JP2013040206A - Method for stimulating motion of digestive system by using ipamorelin - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an effective physiological method for effectively stimulating the motion of the gastrointestinal system.SOLUTION: A therapeutically effective amount of ghrelin mimetic compound represented by structural formula I, its phamaceutically acceptable salt, solvate, or hydrate is administered to a patient.

Description

This application claims the benefit of US Provisional Patent Application No. 61 / 008,828, filed December 21, 2007, the entire contents of which are incorporated herein by reference.
All references (including patents, patent applications, and published patent applications) incorporated herein are incorporated by reference in their entirety, whether or not each is individually cited. .

  Gastrointestinal (GI) motility is a coordinated neuromuscular process that transports nutrients through the digestive system. C. Scarpignato, “Pharmacological Stimulation of Gastrointestinal Motility: Where We Are And Where Are We Going?” Dig. Dis., 15: 112 (1997). Weak (ie, slow) movements of the gastrointestinal system can cause gastroesophageal reflux disease, gastric failure paralysis (eg, diabetic and postoperative paralysis), irritable bowel syndrome, ileus, and constipation (eg, diet or It is one of the biggest health care burdens in industrialized countries. S. D. Feighner et al., “Receptor for Motilin Identified in the Human Gastrointestinal System”, Science, 284: 2184-2188 (June 25, 1999).

  Growth hormone secretagogues (GHS), such as ghrelin and mimetics thereof, have been reported to stimulate gastrointestinal motility. However, certain GHS compounds that have been studied have pharmacokinetic properties that would not be clinically available for the treatment of gastrointestinal motility. In particular, ghrelin is a 28 amino acid peptide produced in the stomach. Biologically active forms of ghrelin (ie acylated forms) have a serum half-life of only 9-13 minutes (Akamizu et al. (2004) European Journal of Endocrinology 150: 447-55). In addition, synthetic GHS compounds such as GHRP-6 have a short serum half-life, which prevents this compound from being used to treat GI motility disorders. Bowers et al. Have shown that the serum half-life of GHRP-6 is only 20 minutes ((1992) Journal of Clinical Endocrinology and Metabolism 74: 292-8).

C. Scarpignato, “Pharmacological Stimulation of Gastrointestinal Motility: Where We Are And Where Are We Going?” Dig. Dis., 15: 112 (1997) S. D. Feighner et al., “Receptor for Motilin Identified in the Human Gastrointestinal System”, Science, 284: 2184-2188 (June 25, 1999) Akamizu et al. (2004) European Journal of Endocrinology 150: 447-55 Journal of Clinical Endocrinology and Metabolism 74: 292-8 (1992)

  Because of the above, an effective physiological method for effectively stimulating gastrointestinal motility is highly desirable and would be an advance in the art.

  The present invention relates to a method of stimulating movement in a patient in need of a method of stimulating gastrointestinal motility, wherein the patient is treated with a gastrointestinal disease (ie, an abnormality, disease, medical condition, or drug or The present invention relates to a method characterized by suffering from a malfunction induced by surgery. This method comprises administering to a patient in need thereof a therapeutically effective amount of a ghrelin mimetic compound or a pharmaceutically acceptable salt, hydrate or solvate thereof. In a preferred embodiment, the ghrelin mimetic is ipamorelin represented by formula I (see below) or a pharmaceutically acceptable salt, hydrate or solvate thereof.

  As mentioned above, ghrelin and ghrelin mimetic compounds have been shown to have a limited serum half-life and are therefore not suitable for use in the treatment of GI motility disorders. For ghrelin and GHRP-6, the serum half-life of ipamorelin has been shown to be 3 to 6.5 hours in humans. Accordingly, the present invention provides methods and therapeutically effective compositions for stimulating gastrointestinal motility.

  Gastrointestinal motility stimulation is used in patients in need of therapeutically effective doses in the treatment of opioid-induced gastrointestinal dysfunction such as morphine-induced colon dysfunction or constipation. Administering a ghrelin mimetic compound or a pharmaceutically acceptable salt, hydrate or solvate thereof. Patients may be using opiate derivatives or opioids for post-surgical pain management or chronic pain management. Typical opiate derivatives and opioids include morphine, codeine, oxycodone, hydromorphone, hydrocodone, methadone, fentanyl, and combinations with anti-inflammatory agents such as acetaminophen or aspirin. A preferred ghrelin mimetic is ipamorelin represented by formula I, or a pharmaceutically acceptable salt, hydrate or solvate thereof.

  Gastrointestinal motility stimulation can be exploited by administering a therapeutically effective amount of a ghrelin mimetic compound in a patient in need thereof to treat gastric paresis. A suitable ghrelin mimetic is ipamorelin represented by formula I, or a pharmaceutically acceptable salt, hydrate, derivative, acid amide or solvate thereof.

  In further embodiments, stimulation of gastrointestinal motility is utilized in a method of treating gastroesophageal reflux disease (GERD) by administering a therapeutically effective amount of a ghrelin mimetic compound in a patient in need thereof. can do. In certain embodiments, the ghrelin mimetic is ipamorelin represented by Formula I, or a pharmaceutically acceptable salt, hydrate or solvate thereof. In certain embodiments, the gastroesophageal reflux disease is a nocturnal gastroesophageal reflux disease.

  The invention also provides a method of stimulating gastrointestinal motility by administering a therapeutically effective amount of a ghrelin mimetic compound in a patient in need thereof for the treatment of irritable bowel syndrome (IBS). provide. A preferred ghrelin mimetic is ipamorelin represented by formula I, or a pharmaceutically acceptable salt, hydrate or solvate thereof. The irritable bowel syndrome may be constipation-primary irritable bowel syndrome or may be alternating constipation / diarrhea irritable bowel syndrome.

  The invention also provides a method of stimulating gastrointestinal motility in a patient in need thereof for treating constipation by administering a therapeutically effective amount of a ghrelin mimetic compound. A preferred ghrelin mimetic is ipamorelin represented by formula I, or a pharmaceutically acceptable salt, hydrate or solvate thereof.

  In one embodiment, stimulation of gastrointestinal motility is therapeutically effective in a patient in need thereof in a method of treating gastrointestinal dysfunction (eg, postoperative ileus) induced by or in connection with surgery. It is utilized by administering an amount of ghrelin mimetic compound. In certain embodiments, the ghrelin mimetic is ipamorelin represented by Formula I, or a pharmaceutically acceptable salt, hydrate or solvate thereof.

又は、製薬上許容しうるその塩、水和物若しくは溶媒和物である。 Suitable ghrelin mimetics are ipamorelin (α-methylalanine-L-histidine-D-β- (2-naphthyl) -alanine-D-phenylalanine-L-lysine amide or H-Aib- represented by Formula I below: His-β- (2-naphthyl) -D-Ala-D-Phe-Lys-NH ₂ ):

Or a pharmaceutically acceptable salt, hydrate or solvate thereof.

  In this disclosure and particularly in the claims and / or paragraphs, the terms “include”, “include”, “include”, etc. may have the meaning ascribed to them in US patent law; Means “includes”, “included”, “includes” and the like; and the terms “consisting essentially of” and “consisting essentially of” In which they recognize components that are not explicitly listed, but that are found in the prior art or that affect basic or novel features of the invention It is noted that the element is excluded.

  These and other embodiments are disclosed in, or are obvious from, and encompassed by the following detailed description.

  The following detailed description, which is given for purposes of illustration and is not intended to limit the invention to only the specific embodiments described, is best understood by reference to the following drawings Can do.

FIG. 4 shows the effect on gastrokinetics of oral administration of 10 and 100 mg / kg ipamorelin in a rat model of postoperative ileus. FIG. 5 shows the efficacy of intravenously administered 0.1, 0.25 or 1.0 mg / kg ipamorelin in a rat model of postoperative ileus. FIG. 4 shows the efficacy of intravenously administered 0.01, 0.03 and 0.1 mg / kg ipamorelin in a rat model of postoperative ileus. Figure 2 shows that ghrelin did not accelerate gastric emptying in rats given 4 mg / kg morphine and that rats given saline or ghrelin showed the same group mean absorbance of phenol red It is. 10 mg / kg RC-1139 administered intravenously with 0.25, 1.0, 2.5 mg / kg ipamorelin (denoted in this figure as 26-0161) to treat postoperative ileus in a rat model It is the figure which showed the effect on gastric motility compared with. FIG. 5 shows reversal of morphine-induced reduction in gastrointestinal motility in humans with ipamorelin alone administered at doses of 0.01, 0.03 and 0.06 mg / kg. 2 is a bar graph comparing the effects of various ghrelin mimetics on gastric emptying. 2 is a bar graph comparing the effects of various ghrelin mimetics on gastrointestinal motility through the small intestine. 2 is a bar graph comparing the effects of various ghrelin mimetics on gastric emptying. 2 is a bar graph comparing the effects of various ghrelin mimetics on gastrointestinal motility through the small intestine (distal from the pyloric sphincter). 2 is a bar graph comparing the effects of various ghrelin mimetics on gastrointestinal motility through the small intestine (proximal to pyloric sphincter). 12A to 12D are diagrams showing the effects of abdominal surgery on colon transit time, fecal excretion, food intake, and weight gain in a rat model of postoperative ileus. FIG. 6 shows colon transit time in rats after abdominal surgery compared to vehicle and control (GHRP-6) after single administration of 0.1 and 1 mg / kg ipamorelin. Cumulative feces compared to vehicle and standard control (GHRP-6) at 12, 24 or 48 hours in rats after abdominal surgery after administration of 0.1 and 1 mg / kg ipamorelin alone It is the figure which showed the grain discharge amount. Cumulative food intake at various time points from 3 to 48 hours after abdominal surgery, after administration of 0.1 and 1 mg / kg ipamorelin alone, with vehicle and standard control (GHRP-6). It is the figure shown in comparison. The figure shows the effect on body weight at 24 and 48 hours after abdominal surgery after administration of 0.1 and 1 mg / kg ipamorelin alone compared to vehicle and standard control (GHRP-6). It is. FIG. 4 shows the colon transit time of rats after abdominal surgery after multiple administrations of 0.01, 0.1 and 1.0 mg / kg ipamorelin compared to vehicle. Diagram showing the effect of 48 hours of fecal excretion compared to vehicle in rats after abdominal surgery after multiple administrations of 0.01, 0.1 and 1.0 mg / kg ipamorelin It is. Showed the effect of cumulative food intake over 48 hours compared to vehicle in rats after abdominal surgery after multiple doses of 0.01, 0.1 and 1.0 mg / kg ipamorelin FIG. FIG. 5 shows the effect of 0.01, 0.1 and 1.0 mg / kg ipamorelin on weight gain over 48 hours in rats after abdominal surgery after multiple doses.

  The present invention relates to a method for stimulating gastrointestinal motility in a patient in need of stimulating gastrointestinal motility, wherein the patient has a gastrointestinal tract disease (ie abnormal or illness or drug or surgery). The present invention relates to the method characterized by suffering from a dysfunction induced by In certain embodiments, these diseases include opioid-induced gastrointestinal dysfunction, such as morphine-induced gastrointestinal dysfunction, constipation, diabetes-related gastric failure, gastroesophageal reflux disease (GERD), Irritable bowel syndrome (IBS) or gastrointestinal dysfunction induced by drugs or surgery, such as postoperative ileus. This method comprises administering to a patient in need thereof a therapeutically effective amount of a ghrelin mimetic compound or a pharmaceutically acceptable salt, hydrate or solvate thereof. The ghrelin mimetic is preferably ipamorelin represented by formula I, or a pharmaceutically acceptable salt, hydrate or solvate thereof.

Ghrelin Mimetics As used herein, the term “ghrelin mimetic” or “ghrelin mimetic compound” or “ghrelin agonist” is the historical term “growth hormone secretagogue” or “growth hormone secretagogue compound” that has been conventionally employed. Is a synonym. A ghrelin mimetic or ghrelin agonist refers to a substance (eg, molecule, compound) that promotes (induces or enhances) at least one function characterized by binding to the ghrelin receptor (GRLN). The GRLN receptor has been previously reported in the literature as a GHS _1a receptor that reflects its first known attribute-growth hormone secretion. This ghrelin receptor is mainly expressed in the hypothalamus and pituitary gland. Activation in the pituitary gland of these receptors induces growth hormone secretion. In addition to inducing growth hormone secretion, recent studies have shown that ghrelin mimetics can increase appetite and weight. In certain embodiments, these ghrelin mimetics are described in US Pat. Nos. 5,767,085, 6,303,620, 6,576,648, 5,977,178, 6,566,337. 6,083,908, 6,274,584, and 6,919,315 (the contents of all of which are incorporated herein by reference).

  Subsequently, this GRLN receptor was identified in locations in the body other than the pituitary and hypothalamus, such as the gastrointestinal tract and vasculature. The binding of ghrelin or ghrelin mimetics to their receptors resulted in pharmacological activities other than or in addition to growth hormone secretion. In particular, other pharmacological activities were increased gastrointestinal prokinetic activity and altered cardio function. Thus, substances previously referred to as growth hormone secretagogue compounds are now more commonly referred to as ghrelin mimetics to represent a broader range of bioactive effects resulting from binding to its receptor (GRLN). Or it is called an agonist.

  Most identified ghrelin mimetics have core peptide backbones of various lengths (tri, tetra, penta, and hexa peptides, and macrocycles). It is also anticipated that different molecular structures will give rise to different affinities for the ghrelin receptor and therefore different pharmacological consequences. It cannot be determined a priori which molecules may have extraordinary activity or efficacy compared to others in the general class. It is generally apparent from specific studies with unusual results or discoveries.

  Compounds having GRLN receptor agonist activity can be identified and evaluated for activity by any suitable method. For example, the binding affinity of a GRLN receptor agonist to the GRRLN receptor can be measured using a receptor binding assay, and growth hormone stimulation is described in US Pat. No. 6,919,315 (for reference, see herein). (Incorporated) can be evaluated. These ghrelin mimetics can be obtained from any source including any commercial supplier.

又は、製薬上許容しうるその塩、水和物、酸、アミド、結晶又は溶媒和物である。 In the case of this invention, suitable ghrelin mimetics are those of the following structural formula I (α-methylalanine-L-histidine-D-β- (2-naphthyl) -alanine-D-phenylalanine-L-lysine amide or H -Ipamorelin represented by -Aib-His-β- (2-naphthyl) -D-Ala-D-Phe-Lys-NH ₂ ):

Or a pharmaceutically acceptable salt, hydrate, acid, amide, crystal or solvate thereof.

  The present invention is based, in part, on the surprising discovery made by the inventor that certain ghrelin mimetics, particularly ipamorelin, have a surprisingly effective and powerful stimulatory effect on gastrointestinal motility. Ipamorelin is a potent growth hormone secretagogue, but its binding affinity with the GRLN receptor is about 2-3 logs weaker than many other reported ghrelin mimetics.

Substances to be co-administered Another aspect of the present invention relates to the co-administration of at least one substance and a ghrelin mimetic, such as ipamorelin, for the treatment of gastrointestinal abnormalities, diseases or conditions. Co-administered can mean that two or more substances are administered together as a single pharmaceutical composition, or that two or more substances are administered in a short time, for example within seconds to days.

Peripheral-acting opioid antagonists Peripheral-acting opioid receptor antagonists such as methylnaltrexone, naltoxone, naltrexone, nalmefene and albimopan (ENTEREG ™) that do not cross the blood-brain barrier, induce side effects induced by opioids, induce opioid withdrawal symptoms Or it can be administered to treat without loss of analgesia. (Holzer P., “Opioids and Opioid Receptors in the Enteric Nervous System: From a Problem in Opioid Analgesia to a Possible New Prokinetic Therapy in Humans” Neurosci Lett., 361 (1-3): 192-5 (2004)) ( Incorporated herein by reference). As used herein, peripherally acting opioid antagonists refer to opioid antagonists that act in the periphery (ie, not the central, eg, the central nervous system).

Proton Pump Inhibitors In another aspect, the present invention provides for co-administration of a ghrelin mimetic such as ipamorelin and a proton pump inhibitor for the treatment of gastrointestinal conditions or diseases. Proton pump inhibitors suppress gastric acid secretion (the final process of acid production) by specific inhibition of the H ⁺ K ⁺ -ATPase enzyme system at the secretory surface of gastric wall cells. Proton pump inhibitors include benzimidazole compounds such as esomeprazole (NEXIUM ™), omeprazole (PRILOSEC ™), lansoprazole (PREVACID ™), rabeprazole (ACIPHEX ™) and pantoprazole (Protonix ™) )including. These proton pump inhibitors contain a sulfinyl group located between the substituted benzimidazole and the pyridine ring. At neutral pH, esomeprazole, omeprazole, lansoprazole, and pantoprazole are chemically stable, lipophilic, weakly basic and have no inhibitory activity. These uncharged weak bases reach the mural cells from the blood and diffuse into the secretory tubules where they are protonated and thereby captured. Protonated species rearrange to form sulfenic acid and sulfenamide, the latter species can interact with the sulfhydryl group of H ⁺ K ⁺ -ATPase. Complete inhibition occurs with two molecules of inhibitor per enzyme molecule. The specificity of the effect of proton pump inhibitors is: a) selective distribution of H ⁺ K ⁺ -ATPase; b) that acidic conditions are required to catalyze the formation of reactive inhibitors; and c) in acidic tubules It is believed to be derived from the capture of protonated drugs and cationic sulfenamides at and adjacent to the target enzyme. Goodman &Gilman's The Pharmacological Basis of Therapeutics, 9th edition, 901-915 (1996) (incorporated herein by reference).

H ₂ receptor antagonists In yet another aspect, the invention provides for the simultaneous administration of a ghrelin mimetic, eg, ipamorelin, and an H ₂ receptor antagonist for the treatment of a gastrointestinal condition or disease. H ₂ receptor antagonists competitively inhibit the interaction of histamine with the H ₂ receptor. They are highly selective and have little or no effect on the H ₁ receptor. Even though H ₂ receptors are present in many tissues, including blood vessels and bronchial smooth muscle, H ₂ receptor antagonists have little noticeable intervention in physiological functions other than gastric acid secretion. H ₂ receptor antagonists include nizatidine (AXID ™), ranitidine (ZANTAC ™ and TRITEC ™), famotidine (PEPCCID AC ™), and cimetidine (TAGAMET ™). Goodman &Gilman's The Pharmacological Basis of Therapeutics, 9th edition, pages 901-915 (1996)) (incorporated herein by reference). H ₂ receptor antagonists inhibit gastric acid secretion elicited by histamine, other H ₂ agonists, gastrin (and to a lesser extent muscarinic agonists). H ₂ receptor antagonists also inhibit basal and nocturnal acid secretion.

Antacids Another aspect of the invention provides a method of co-administration of a ghrelin mimetic such as ipamorelin and an antacid for the treatment of a gastrointestinal condition or disease. For example, the compounds of this invention can be co-administered with an antacid to neutralize gastric acid. For example, aluminum hydroxide and magnesium (MAALOX ™ and MYLANTA ™) neutralize gastric acid and increase the pH in the stomach and duodenal bulb.

Laxatives The present invention further provides a method for co-administering a ghrelin mimetic such as ipamorelin and a laxative to treat gastrointestinal conditions or diseases. Laxatives may take a variety of forms including, for example, liquids, tablets, suppositories, powders, granules, capsules, chewing gum, chocolate flavored wafers, and caramel. The basic types of laxatives are expansive laxatives, lubricious laxatives, stool softeners (also called mild laxatives), and irritant laxatives.

  Intumescent laxatives include materials such as cellulose and psyllium that pass undigested through the digestive tract. For example, these materials absorb liquid and swell, making the stool soft and bulky to facilitate passage. This bulky stool then stimulates the colon to exercise. This group of laxatives includes brands such as FIBERCON ™, FIBERALL ™, and METAMUCIL ™.

  Lubricating laxatives include, for example, mineral oil. Mineral oil is the most widely used lubricant laxative. If taken by mouth, the oil coats the stool. This keeps the stool moisture and soft and facilitates its passage. Lubricating laxatives are often used in patients who need to avoid a stomach rash (eg, after abdominal surgery).

  Stool softeners (relaxing laxatives) make stools softer and easier to pass by increasing their moisture content. This type of laxative does not actually stimulate colonic movement, but allows colonic movement without bloating. Stool softeners are best used, for example, to prevent constipation in people who need to avoid bloating due to recent surgery. Fecal softeners include, for example, doxate sodium (COLACE ™, REGUTOL ™, etc.), doxate calcium (SURFAK ™, DC SOFTGELS ™) and doxate potassium (DIALOSE ™), DIOCTO-K ™).

Serotonin receptor (5-HT) agonist (pure or mixed)
The present invention also ghrelin mimetic example ipamorelin with serotonin receptor agonists for example 5-HT ₄ agonist, also provides a method for co-administration in order to treat the condition or disease of the gastrointestinal. These serotonin agonists may be pure or mixed 5-HT receptor subtypes, or may be a mixture with other central nervous system receptors such as dopamine.

5-HT ₄ agonists speed up the movement of colon contents through the colon and reduce sensitivity to enteric nerve stimulation. Suitable serotonin agonists that can be used in combination with the compounds of this invention include, but are not limited to, laurucine, yohimbine, metoclopramide, purcarpride and tegaserod (ZELNORM ™). Spiller R., “Serotonergic Modulating Drugs for Functional Gastrointestinal Diseases”, Br J Clin Pharmacol. 54: 11-20 (2002) and US Pat. No. 6,413,988 (incorporated herein by reference).

The Motilin Receptor Agonist The present invention further provides a method for the simultaneous administration of a ghrelin mimetic such as ipamorelin and a motilin receptor agonist for the treatment of a gastrointestinal condition or disease. Motilin is a 22 amino acid peptide produced in the gastrointestinal tract system of many species. Motilin induces smooth muscle contraction in dog, rabbit and human stomach tissue and rabbit colon. Apart from local gastrointestinal tissue, motilin and its receptor have been found in other tissues.

  In addition to motilin, there are other substances that are agonists of the motilin receptor and induce excretion of gastrointestinal contents. One of those drugs is the antibiotic erythromycin. Studies have shown that erythromycin elicits a biological response comparable to motilin itself and therefore may be useful in the treatment of diseases such as chronic idiopathic simulated bowel obstruction and gastric paralysis. Weber, F. et al., The American Journal of Gastroenterology, 88: 4, 485-90 (1993) (incorporated herein by reference).

Dopamine Antagonists Another aspect of the present invention provides methods for the simultaneous administration of ghrelin mimetics such as ipamorelin and dopamine antagonists to treat gastrointestinal conditions or diseases.

  Dopamine antagonists are drugs that bind to dopamine receptors but do not activate them, thereby blocking the action of dopamine or exogenous agonists. This class of drugs includes, but is not limited to, metoclopramide, domperidone, amisulpride, cleboprid, mosapramine, nemonapride, remoxiprid, risperidone, sulpiride, sultopride and ziprasidone.

Cholinesterase Inhibitors The present invention also provides methods for co-administering a ghrelin mimetic such as ipamorelin and a cholinesterase inhibitor to treat a gastrointestinal condition or disease. The term “cholinesterase inhibitor” refers to at least one agent that prolongs the action of acetylcholine by inhibiting its destruction or hydrolysis by cholinesterase. Cholinesterase inhibitors are also known as acetylcholinesterase inhibitors. Examples of cholinesterase inhibitors include edrophonium, neostigmine, neostigmine methyl sulfate, pyridostigmine, tacrine and physostigmine, ambenonium chloride (MYTEASE ™), edrophonium chloride (TENSILON ™), neostigmine (PROSTIGMIN ™) , Pyridogustimina (MESTINON ™), distigmine bromide, eptastigmine, galantamine, acecridine, acetyl chlorine bromide, acetyl chlorin chloride, acratonium napadisilate, benzpyrinium bromide, carbachol, carbonium chloride, ceme potassium bromide, dexpan Tenol, diisopropyl paraoxon, ecothiofetochloride, ethanol Lysine, Frankfurt Leto chloride, Metako link chloride de, muscarinic, oxa prop iodide, and include xanomeline, not limited to these.

Stereochemistry Many of the compounds described herein have at least one chiral center and can therefore exist in various mirror image types. If desired, the chiral carbon can be indicated with an asterisk ( ^* ). Where the bond to the chiral carbon is depicted as a straight line in the formula of this invention, both the (R) and (S) configurations of the chiral carbon are, therefore, both enantiomers and mixtures thereof are represented by the formula It is understood that they are encompassed within. When used in the field, if one wishes to specify the absolute configuration of a chiral carbon, one of the bonds to the chiral carbon can be depicted as a wedge (bond to the upper atom on the page) and another Can be drawn as a series of short parallel lines or wedges (bonds to atoms below the plane of the paper). Using the Khan-Ingold-Prelog system, (R) or (S) can be assigned to a chiral carbon.

  When a compound of the present invention has more than one chiral carbon, it can have more than one optical isomer and can exist as diastereoisomers. For example, if there are two chiral carbons, the compound can have up to four optical isomers and two pairs of enantiomers ((S, S) / (R, R) and (R, S) / ( S, R)). These enantiomeric pairs (eg, (S, S) / (R, R)) are mirror image stereoisomers of each other. Stereoisomers that are not mirror images (eg, (S, S) and (R, S)) are diastereoisomers. These diastereoisomer pairs can be separated by methods known to those skilled in the art, such as chromatography or crystallization, and the individual enantiomers in each pair can be separated as described above. The present invention includes each diastereoisomer of such compounds and mixtures thereof.

Screening It is understood that ghrelin mimetic compounds can be identified, for example, by screening a library or collection of molecules using an appropriate method. Another source of compounds of interest is a combinatorial library that can contain many structurally distinct molecular species. Combinatorial libraries can be used to identify lead compounds or to optimize previously identified lead compounds. Such libraries can be produced by well-known methods of combinatorial chemistry and screened by appropriate methods.

Method of Treating Gastrointestinal Diseases The present invention is a method of stimulating gastrointestinal motility in a patient in need thereof, wherein the patient has a gastrointestinal disease (ie, illness, abnormality, medical condition, or drug or Provided are such methods suffering from surgery-induced dysfunction. This method comprises administering to a patient in need thereof a therapeutically effective amount of a ghrelin mimetic compound or a pharmaceutically acceptable salt, hydrate or solvate thereof. The ghrelin mimetic is ipamorelin represented by Formula I, or a pharmaceutically acceptable salt, hydrate or solvate thereof.

  As used herein, the term “gastrointestinal disease” refers to any disease, abnormality, condition, or dysfunction that results in impaired gastrointestinal function. For example, the gastrointestinal disorder may be opioid-induced gastrointestinal dysfunction, such as constipation induced by morphine, postoperative ileus, or gastric failure paralysis.

Constipation In another aspect, the invention provides a method of treating constipation by administering a therapeutically effective amount of a ghrelin mimetic, such as ipamorelin. Constipation is a condition with unpleasant or frequent colonic movement. Persons with constipation may have hard stools and have difficulty in defecation. The person can also feel as if the rectum is not completely emptied. Acute constipation suddenly starts prominently. On the other hand, chronic constipation begins instinctively and can last for months or years.

  The method of treating constipation of the present invention can further comprise co-administering a ghrelin mimetic, such as ipamorelin, with a therapeutically effective amount of a laxative. Suitable laxatives include, but are not limited to, inflatable laxatives, lubricious laxatives, stool softeners, or any combination thereof.

Opioid-Induced Constipation This invention provides a method of treating opioid-induced constipation by administering a therapeutically effective amount of a ghrelin mimetic such as ipamorelin. The use of opioid analgesics to relieve chronic pain can cause effects on organs outside the central nervous system target. For example, the action of opioids can slow gastric emptying and inhibit colonic movement. Long-term stool retention in the intestine results in excessive moisture and sodium absorption from the stool contents, which results in harder, drier stool and constipation. This effect afflicts about 90% of individuals using analgesics. For patients with chronic pain who are receiving opioid medication, the resulting constipation can be a side effect that limits dosage. In addition, analgesics used for post-surgical pain management can cause opioid-induced constipation. Suitable opioids include, but are not limited to, morphine, codeine, oxycodone, hydromorphone, hydrocodone, methadone, fentanyl, and anti-inflammatory agents such as acetaminophen or aspirin or any combination thereof. .

  The method of treating opioid-induced constipation can further comprise co-administering a ghrelin mimetic compound, such as ipamorelin, with a therapeutically effective amount of a peripherally acting opioid antagonist, laxative or any combination thereof. Suitable peripherally acting opioid antagonists include, but are not limited to, methylnaltrexone, naltrexone, nalmefene, naloxone and albimopan or any combination thereof. Suitable laxatives include, but are not limited to, inflatable laxatives, lubricious laxatives, and stool softeners, or any combination thereof.

Post-operative ileus The present invention provides a method of treating post-operative ileus by administering a therapeutically effective amount of a ghrelin mimetic such as ipamorelin. It has been well documented that gastrointestinal (GI) tract movement is temporarily impaired after surgery. The impact that abdominal surgery has on gastrointestinal motility is commonly referred to as “post-operative ileus”, a term that refers to normal stressed movement disruption of the gastrointestinal tract. Peristaltic movement becomes dysfunctional. Ileus is also defined as a functional, non-mechanical disorder of the colon. The term “postoperative ileus” refers to a delay in draining contents from the normal stomach and intestines.

  The method of treating post-operative ileus can further comprise co-administering a ghrelin mimetic, such as ipamorelin, with a therapeutically effective amount of a dopamine antagonist. Suitable dopamine antagonists include, but are not limited to, metoclopramide, domperidone, amisulpride, clevopride, mosapramine, nemonapride, remoxiprid, risperidone, sulpiride, sultopride and ziprasidone, or any combination thereof.

Irritable Colon Syndrome The present invention provides a method of treating irritable bowel syndrome by administering a therapeutically effective amount of a ghrelin mimetic such as ipamorelin. Irritable bowel syndrome (IBS) is a dysfunction that affects the movement of the entire gastrointestinal tract and can cause abdominal pain, constipation, and / or diarrhea. Impaired gastrointestinal motility in IBS is not accompanied by changes in physical structure such as inflammation or tumor. IBS symptoms are thought to be related to abnormal muscle contraction in any part of the intestine.

  In this syndrome, the gastrointestinal tract is particularly sensitive to gastrointestinal irritation. Stress, diet, drugs, hormones, or minor stimuli can cause abnormal contractions of the gastrointestinal tract. There are different types of IBS: constipation-dominated IBS, diarrhea-dominated IBS, and IBS with alternating constipation and constipation dominance.

A method of treating IBS can comprise co-administering a ghrelin mimetic compound, such as ipamorelin, with a therapeutically effective amount of an H ₂ receptor antagonist; a serotonin 5-HT agonist; a laxative; or any combination thereof.

Suitable H ₂ receptor antagonists include but are not limited to nizatidine, ranitidine, famotidine, and cimetidine, or any combination thereof. Suitable central nervous system receptor agonists include, but are not limited to, laurucine, yohimbine, metoclopramide, tegaserod, or any combination thereof. Suitable laxatives include, but are not limited to, inflatable laxatives, lubricating laxatives, stool softeners, or any combination thereof.

Gastroesophageal Reflux Disease This invention further provides a method of treating gastroesophageal reflux disease by administering a therapeutically effective amount of a ghrelin mimetic such as ipamorelin. Gastroesophageal reflux disease (GERD) is a condition in which stomach contents (eg, bile salts) return to the esophagus, causing chronic reflux of the stomach contents from the stomach to the lower esophagus. GERD, commonly known as heartburn, causes esophageal irritation and inflammation.

  In people with GERD, the esophageal sphincter (the circular muscle located at the lower end of the esophagus to prevent stomach contents from returning to the esophagus) cannot perform its protective mission. Instead of opening only when a person eats or drinks, it relaxes and allows the digestive fluid to flow back into the esophagus and irritate the lining of the esophagus.

  Two types of GERD have been identified, upright or daytime GERD and supine or nighttime GERD. The occurrence of nocturnal reflux symptoms is infrequent, but the acid clearance is longer. Nocturnal reflux can be combined with GERD complications such as esophageal fistulas, ulcers and respiratory symptoms. Currently, it is estimated that 17 million Americans suffer from heartburn and other symptoms of GERD.

A method of treating GERD includes co-administering a ghrelin mimetic compound, such as ipamorelin, with a therapeutically effective amount of an H ₂ receptor antagonist; an antacid; a proton pump inhibitor; or any combination thereof.

Suitable H ₂ receptor antagonists include but are not limited to nizatidine, ranitidine, famotidine, and cimetidine, or any combination thereof. Suitable antacids include, but are not limited to aluminum hydroxide and magnesium hydroxide and combinations thereof. Suitable proton pump inhibitors include, but are not limited to, esomeprazole (NEXIUM ™), omeprazole, lansoprazole, pantoprazole, or combinations thereof.

Gastroparesis The present invention provides a method of treating gastric failure, eg, diabetic or idiopathic gastric failure, by administering a therapeutically effective amount of a ghrelin mimetic, eg, ipamorelin. Gastroparesis (also called slow emptying of the stomach contents) is an abnormality that takes too long for the stomach to drain its contents. It often occurs in people with type 1 diabetes or type 2 diabetes. Gastroparesis can occur when nerves to the stomach are damaged or stop functioning. The vagus nerve controls the movement of food through the digestive tract. If the vagus nerve is damaged, stomach and intestinal muscles do not work properly and food movements are slowed or stopped. Diabetes can damage the vagus nerve if blood glucose levels remain high over time. High blood glucose causes chemical changes in the nerves and damages the blood vessels that carry oxygen and nutrients to the nerves.

  The method of treating gastric paresis may comprise co-administering a ghrelin mimetic compound, such as ipamorelin, with a therapeutically effective amount of a dopamine antagonist. Suitable dopamine antagonists include, but are not limited to, metoclopramide, domperidone, amisulpride, clevopride, mosapramine, nemonapride, remoxiprid, risperidone, sulpiride, sultopride and ziprasidone, or any combination thereof.

  The invention further refers to pharmaceutical compositions useful for stimulating (ie, inducing) gastrointestinal motility. The pharmaceutical composition comprises a ghrelin mimetic and an optional pharmaceutically acceptable carrier. The pharmaceutical composition can include a second amount of a suitable therapeutic agent. An appropriate therapeutic agent can be determined based on the condition of the patient being treated.

  For example, the pharmaceutical composition can include a first amount of a ghrelin mimetic, such as ipamorelin, and a second amount of a laxative (when treating constipation). The pharmaceutical composition of the present invention can appropriately contain a pharmaceutically acceptable carrier. The ghrelin mimetic and laxative can each be present in the pharmaceutical composition in a therapeutically effective amount. In other aspects, the first and second amounts can together constitute a therapeutically effective amount.

The pharmaceutical composition can comprise a first amount of a ghrelin mimetic and a second amount of an H ₂ receptor antagonist. The pharmaceutical composition of the present invention can appropriately contain a pharmaceutically acceptable carrier. The ghrelin mimetic and H ₂ receptor antagonist can each be present in the pharmaceutical composition in a therapeutically effective amount. In other aspects, the first and second amounts can together constitute a therapeutically effective amount.

  The pharmaceutical composition can comprise a first amount of a ghrelin mimetic and a second amount of a serotonin receptor agonist. The pharmaceutical composition can optionally contain a pharmaceutically acceptable carrier. The ghrelin mimetic and serotonin receptor agonist can each be present in the pharmaceutical composition in a therapeutically effective amount. The first and second amounts together can constitute a therapeutically effective amount.

  The pharmaceutical composition can include a first amount of a ghrelin mimetic such as ipamorelin and a second amount of an antacid. The pharmaceutical composition can optionally contain a pharmaceutically acceptable carrier. The ghrelin mimetic and antacid can each be present in the pharmaceutical composition in a therapeutically effective amount. In other aspects, the first and second amounts together can constitute a therapeutically effective amount.

  The pharmaceutical composition can include a first amount of a ghrelin mimetic and a second amount of an opioid antagonist. The pharmaceutical composition can optionally contain a pharmaceutically acceptable carrier. The ghrelin mimetic and opioid antagonist can each be present in the pharmaceutical composition in a therapeutically effective amount. The first and second amounts together can constitute a therapeutically effective amount.

  The pharmaceutical composition can comprise a first amount of a ghrelin mimetic and a second amount of a proton pump inhibitor. The pharmaceutical composition can optionally contain a pharmaceutically acceptable carrier. The ghrelin mimetic and proton pump inhibitor can each be present in the pharmaceutical composition in a therapeutically effective amount. The first amount and the second amount together can constitute a therapeutically effective amount.

  The pharmaceutical composition can comprise a first amount of a ghrelin mimetic and a second amount of a motilin receptor agonist. The pharmaceutical composition can optionally contain a pharmaceutically acceptable carrier. The ghrelin mimetic and motilin receptor agonist can each be present in the pharmaceutical composition in a therapeutically effective amount. The first and second amounts together can constitute a therapeutically effective amount.

  The pharmaceutical composition can comprise a first amount of a ghrelin mimetic and a second amount of a dopamine antagonist. The medicament can optionally contain a pharmaceutically acceptable carrier. The ghrelin mimetic and dopamine antagonist can each be present in the pharmaceutical composition in a therapeutically effective amount. The first and second amounts together can constitute a therapeutically effective amount.

  The pharmaceutical composition can comprise a first amount of a ghrelin mimetic and a second amount of a cholinesterase inhibitor. The pharmaceutical composition can optionally contain a pharmaceutically acceptable carrier. The ghrelin mimetic and cholinesterase inhibitor can each be present in the pharmaceutical composition in a therapeutically effective amount. The first and second amounts together can constitute a therapeutically effective amount.

  The pharmaceutical composition can comprise a first amount of ghrelin mimetic and a second amount of somatostatin. The pharmaceutical composition can optionally contain a pharmaceutically acceptable carrier. The ghrelin mimetic and somatostatin can each be present in the pharmaceutical composition in a therapeutically effective amount. The first and second amounts together can constitute a therapeutically effective amount.

  This invention further refers to the use of ghrelin mimetic compounds for the manufacture of a medicament for stimulating (ie, inducing) gastrointestinal motility.

  Patient, as used herein, refers to an animal, such as a mammal, and is a primate (eg, human), cow, sheep, goat, horse, pig, dog, cat, rabbit, guinea pig, rat, mouse or other bovine genus , Sheep-like animals, equine, canine, feline, rodent or murine species. In preferred embodiments, the mammal is a human.

  As used herein, treating and treating refers to stimulating (eg, inducing) gastrointestinal motility.

  As used herein, a therapeutically effective amount refers to an amount sufficient to elicit a desired biological response. The desired biological response is to stimulate (eg, induce) gastrointestinal motility. The desired biological response is to stimulate (eg, induce) gastrointestinal motility in patients in need thereof to treat opioid-induced constipation. The patient may be one who is using opioids for postoperative pain management or for chronic pain management.

  The desired biological response is to stimulate (eg, induce) gastrointestinal motility in patients in need thereof to treat gastric paresis.

  The desired biological response is to stimulate (eg, induce) gastrointestinal motility in a patient in need thereof to treat gastroesophageal reflux disease. This gastroesophageal reflux disease is a nighttime gastroesophageal reflux disease.

  The desired biological response is to stimulate (eg, induce) gastrointestinal motility in a patient in need thereof to treat irritable bowel syndrome. This irritable bowel syndrome is constipation-dominated irritable bowel syndrome. In yet another embodiment, the irritable bowel syndrome is constipation / diarrhea irritable bowel syndrome.

  The desired biological response is to stimulate (eg, induce) gastrointestinal motility in patients in need thereof to treat constipation.

  The desired biological response is to stimulate (eg, induce) gastrointestinal motility in a patient in need thereof to treat postoperative ileus.

Pharmaceutical Compositions Pharmaceutical compositions suitable for use in the present invention include compositions containing the active ingredients in a therapeutically effective amount to achieve the intended purpose. The unit content of the active ingredient contained in the individual dosage of each dosage form does not have to constitute an effective amount on its own, which means that an effective amount required can be a plurality of dosage units (capsules or tablets). Or it may be achieved by administration of vials or combinations thereof. In addition, it is understood that at some dosage levels, an effective amount may not show any measurable effect until after one week, one month, three months, or six months of use. Determination of an effective amount is well within the capability of those skilled in the art, particularly within the capability of those skilled in the art in light of the detailed disclosure provided herein. The particular dosage level for any particular user will depend on various factors including age, physical activity level, general health status, and severity of gastrointestinal disease.

A therapeutically effective dose also refers to that amount necessary to achieve the desired effect without causing undesirable or unacceptable side effects. Toxicity and therapeutic efficacy of ghrelin mimetics such as ipamorelin of this invention can be measured by standard pharmaceutical procedures in cell cultures or experimental animals. Standard methods can be used to determine the ED ₅₀ , a dosage that shows efficacy in about 50% of the test population. Similarly, the SD ₅₀ , the dosage that produces undesirable side effects in 50% of the population, can be measured. The dose ratio between side effects and therapeutic effects can be expressed as a therapeutic index, which can be expressed as the ratio between SD ₅₀ / ED ₅₀ . A ghrelin mimetic with a high therapeutic index (eg, ipamorelin), ie one that is effective at low dosages and has no undesirable side effects (if at all, only at very high doses) is preferred is there. A suitable treatment index is greater than about 3, more preferably the treatment index is greater than 10, and most preferably the treatment index is greater than 25 (eg, greater than 50). Furthermore, ghrelin mimetics that have no side effects at any dosage level are more preferred. Finally, ghrelin mimetics that are effective at low dosages and have no side effects at any dosage level are most preferred. The exact formulation, route of administration and dosage can be chosen according to the desired effect and can be made by one skilled in the art.

  In certain embodiments, these ghrelin mimetics are formulated as pharmaceutically acceptable salts. As used herein, the term pharmaceutically acceptable salt refers to a salt of a compound to be administered, its solvate, prepared from a pharmaceutically acceptable non-toxic acid (including inorganic and organic acids), Refers to hydrate or clathrate. Examples of such inorganic acids are hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, sulfuric acid and phosphoric acid. Suitable organic acids can be selected from, for example, fatty acids, aromatic acids, carboxylic acids and organic acid classes of sulfonic acids, examples of which include formic acid, acetic acid, propionic acid, succinic acid, camphor sulfonic acid, citric acid. Acid, fumaric acid, gluconic acid, isethionic acid, lactic acid, malic acid, mucoic acid, tartaric acid, para-toluenesulfonic acid, glycolic acid, glucuronic acid, maleic acid, furoic acid, glutamic acid, benzoic acid, anthranilic acid, salicylic acid, phenyl Examples include acetic acid, mandelic acid, embonic acid (pamoic acid), methanesulfonic acid, ethanesulfonic acid, pantothenic acid, benzenesulfonic acid (besylate), stearic acid, sulfanilic acid, alginic acid, galacturonic acid, and the like.

  The ghrelin mimetics of this invention can be made from their hydrates such as hemihydrate, monohydrate, dihydrate, trihydrate, tetrahydrate and the like and solvates.

  Ghrelin mimetics such as ipamorelin and its derivatives and any co-administered drugs can be incorporated into any suitable pharmaceutical composition that may be suitable for administration. Such compositions typically comprise an active agent (eg, a ghrelin mimetic of this invention) and a pharmaceutically acceptable carrier.

  As used herein, “pharmaceutically acceptable carrier” refers to any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like (compatible with pharmaceutical administration). Intended to be included). The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional medium or agent is incompatible with the active compound, their use in these compositions is contemplated. Supplementary active compounds can also be incorporated into the pharmaceutical compositions of the invention. Any of the components of the pharmaceutical composition can be modified to affect the solubility or cleansing value of the factors of the invention. Peptide molecules can also be synthesized with D-amino acids to increase resistance to enzymatic degradation. In some cases, the composition can be co-administered with one or more solubilizers, preservatives, and permeability enhancers.

The therapeutically effective amount or dose of a ghrelin mimetic depends on the patient's age, sex, and weight, and the patient's current health status. One skilled in the art can determine the appropriate dosage according to these and other factors to achieve the desired biological response.

  Suitable daily doses of the ghrelin mimetic of this invention are about 1 ng to about 10,000 mg, about 5 ng to about 9,500 mg, about 10 ng to about 9,000 mg, about 20 ng to about 8,500 mg, about 30 ng. To about 7,500 mg, about 40 ng to about 7,000 mg, about 50 ng to about 6,500 mg, about 100 ng to about 6,000 mg, about 200 ng to about 5,500 mg, about 300 ng to about 5,000 mg, about 400 ng to about 4,500 mg, about 500 ng to about 4,000 mg, about 1 μg to about 3,500 mg, about 5 μg to about 3,000 mg, about 10 μg to about 2,600 mg, about 20 μg to about 2,575 mg, about 30 μg to about 2, 550 mg, about 40 μg to about 2500 mg, about 50 μg to about 2,475 mg, about 100 μg to about 2,450 mg, about 200 μg to 2,425 mg, about 300 μg to about 2,000, about 400 μg to about 1,175 mg, about 500 μg to about 1,150 mg, about 0.5 mg to about 1,125 mg, about 1 mg to about 1,100 mg, about 1.25 mg To about 1,075 mg, about 1.5 mg to about 1,050 mg, about 2.0 mg to about 1,025 mg, about 2.5 mg to about 1,000 mg, about 3.0 mg to about 975 mg, about 3.5 mg to about 950 mg, about 4.0 mg to about 925 mg, about 4.5 mg to about 900 mg, about 5 mg to about 875 mg, about 10 mg to about 850 mg, about 20 mg to about 825 mg, about 30 mg to about 800 mg, about 40 mg to about 775 mg, about 50 mg To about 750 mg, about 100 mg to about 725 mg, about 200 mg to about 700 mg, about 300 mg to about 675 mg, about 400 mg to about 6 0 mg, may be in the range of about 500mg, or about 525mg~ about 625 mg.

  Other suitable daily doses of ghrelin mimetics of this invention are about 1 ng, about 5 ng, about 10 ng, about 20 ng, about 30 ng, about 40 ng, about 50 ng, about 100 ng, about 200 ng, about 300 ng, about 400 ng, about 500 ng, about 1 μg, about 5 μg, about 10 μg, about 20 μg, about 30 μg, about 40 μg, about 50 μg, about 100 μg, about 200 μg, about 300 μg, about 400 μg, about 500 μg (0.5 mg), about 1 mg, about 1.25 mg, about 1.5 mg, about 2.0 mg, about 2.5 mg, about 3.0 mg, about 3.5 mg, about 4.0 mg, about 4.5 mg, about 5 mg, about 10 mg, about 20 mg, about 30 mg , About 40 mg, about 50 mg, about 100 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 625 mg, about 650 m About 675 mg, about 700 mg, about 725 mg, about 750 mg, about 775 mg, about 800 mg, about 825 mg, about 850 mg, about 875 mg, about 900 mg, about 925 mg, about 950 mg, about 975 mg, about 1000 mg, about 1025 mg, about 1050 mg, about 1075 mg, about 1100 mg, about 1125 mg, about 1150 mg, about 1175 mg, about 1200 mg, about 1225 mg, about 1250 mg, about 1275 mg, about 1300 mg, about 1325 mg, about 1350 mg, about 1375 mg, about 1400 mg, about 1425 mg, about 1450 mg, about 1475 mg, About 1500 mg, about 1525 mg, about 1550 mg, about 1575 mg, about 1600 mg, about 1625 mg, about 1650 mg, about 1675 mg, about 1700 mg, about 1725 mg, 1750 mg, about 1775 mg, about 1800 mg, about 1825 mg, about 1850 mg, about 1875 mg, about 1900 mg, about 1925 mg, about 1950 mg, about 1975 mg, about 2000 mg, about 2025 mg, about 2050 mg, about 2075 mg, about 2100 mg, about 2125 mg, about 2150 mg, About 2175 mg, about 2200 mg, about 2225 mg, about 2250 mg, about 2275 mg, about 2300 mg, about 2325 mg, about 2350 mg, about 2375 mg, about 2400 mg, about 2425 mg, about 2450 mg, about 2475 mg, about 2500 mg, about 2525 mg, about 2550 mg, about 2575 mg About 2600 mg, about 3,000 mg, about 3,500 mg, about 4,000 mg, about 4,500 mg, about 5,000 mg, about 5,500 mg, A dosage of about 6,000 mg, about 6,500 mg, about 7,000 mg, about 7,500 mg, about 8,000 mg, about 8,500 mg, about 9,000 mg, or about 9,500 mg or more is included.

  Suitable dosages of ghrelin mimetics may range from about 0.20 mg to about 4000 mg per day, such as from about 1 mg to about 4000 mg, such as from about 5 mg to about 3000 mg, such as from about 10 mg to about 2400 mg. It may be. This dose can be administered in a single dose or in multiple doses (eg, 1 to 4 times per day). When multiple doses are used, each dose may be the same or different.

  Suitable dosages for additional therapeutic agents such as laxatives may be in the same range as described above for ghrelin mimetics. The dosage of the ghrelin mimetic and the additional drug may be the same or different. Appropriate doses of additional agents can be found in the literature.

  The compounds for use in the methods of this invention may be any suitable route such as oral or parenteral administration (eg transdermal, transmucosal (eg sublingual, lingual, (trans) buccal), Vaginal administration (eg transvaginal and pervaginal administration), nasal (intracavitary) and rectal (internal) administration, subcutaneous administration, intramuscular administration, intradermal administration, intraarterial administration, intravenous administration, inhalation and topical administration For administration by administration.

  In preferred embodiments, the compounds of this invention are formulated for intravenous delivery. In other preferred embodiments, the compounds of this invention are formulated for oral delivery. Suitable compositions and dosage forms include tablets, capsules, caplets, pills, gel caps, troches, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pellets, magma Includes lozenges, creams, pastes, salves, lotions, discs, suppositories, liquid sprays, dry powders or aerosolized formulations.

  It is preferred that these compounds be administered orally. Suitable oral dosage forms include, for example, by conventional means, pharmaceutically acceptable excipients such as binders (eg, polyvinylpyrrolidone or hydroxypropylmethylcellulose); fillers (eg, lactose, microcrystalline cellulose or calcium phosphate). Tablets, capsules or caplets made with a lubricant (eg, magnesium stearate, talc or silica); a disintegrant (eg, sodium starch glycolate); or a wetting agent (eg, sodium lauryl sulfate). included. If desired, using suitable methods, these tablets can be coated, for example, to facilitate swallowing or to provide a delayed release of the active ingredient. Liquid preparations for oral administration may be in the form of solutions, syrups or suspensions. Liquid formulations (eg, solutions, suspensions and syrups) are also suitable for oral administration, and pharmaceutically acceptable additives such as suspending agents (eg, sorbitol syrup, methylcellulose or hydrogenated edible fat); Produced by conventional means, with emulsifiers (eg lecithin or gum arabic); non-aqueous vehicles (eg almond oil, oily esters or ethyl alcohol); and preservatives (eg methyl or propyl p-hydroxybenzoate or sorbic acid) can do.

  Preferably, a pharmaceutical composition comprising a ghrelin mimetic of this invention is for treating a gastrointestinal disorder (eg, gastrointestinal tract abnormality, disease, condition or injury that impairs gastrointestinal motility) by stimulating gastrointestinal motility. Used for. The exact formulation, route of administration and dosage can be chosen according to the desired effect and can be made by one skilled in the art.

  Dosage intervals can be determined by experimental testing. One or more ghrelin mimetics of this invention could be administered using a regimen that maintains gastrointestinal motility 10%, about 20%, about 50% above or below normal.

  Another suitable method of administration provides the ghrelin mimetic of this invention, a cell line capable of expressing the ghrelin mimetic by transplant, or the transplant or cell line provides the ghrelin mimetic to cells of the gastrointestinal system Is to give as much as you can.

  A pharmaceutical composition of the invention can be formulated to be compatible with its intended route of administration.

  Oral administration refers to administration of the pharmaceutical composition of this invention by ingestion through the mouth or through any other part of the gastrointestinal system including the esophagus or by suppository administration. Parenteral administration refers to delivery of a composition, eg, a composition comprising a ghrelin mimetic, by a route other than the route through the gastrointestinal tract (eg, oral delivery). In particular, parenteral administration may be intra-intestinal injection, subcutaneous injection, intramuscular injection or intramedullary (ie intrathecal) injection. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic. Topical administration refers to the application of the medicament to the outer surface of the skin or mucous membranes (including the surface membranes of the nose, lungs and mouth (which may also be in the form of oral administration in this case)), so that the drug is applied to the skin Or it enters the underlying tissue across the outer surface of the mucosa. Topical administration of a drug can result in a limited distribution of the drug to the skin and surrounding tissue, or can result in a systemic distribution of the drug if it is moved from the treatment area by the bloodstream .

  In one form of topical administration contemplated by this invention, the ghrelin mimetic is delivered by transdermal delivery. Transdermal delivery refers to the diffusion of a drug across a skin barrier. Absorption through intact skin can be enhanced by placing the active agent in an oily vehicle (a method known as oiling) and the use of microneedles prior to application to the skin. Passive topical administration may consist of applying the active agent directly to the treatment site along with a palliative or penetration enhancer. Another way to enhance delivery through the skin is to increase the dosage of the medicament. Dosages for topical administration can be increased up to 10 times, 100 times or 1000 times the dosage administered by other routes.

  Penetrants for transdermal delivery are generally known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For administration by inhalation, the ghrelin mimetics of this invention may be delivered in the form of an aerosol spray from a pressurized container or wave dispenser containing a gas such as a suitable propellant, such as carbon dioxide, or in a nebulizer. Can do. For transdermal administration, the ghrelin mimetics of this invention can be formulated into ointments, salves, gels, or creams as is generally known in the art.

  In addition, the ghrelin mimetics of this invention can be delivered by nasal or pulmonary administration. Pulmonary delivery of aerosolized medicaments has been described in a number of references, beginning with Gansslen (1925) Klin. Wochenschr. 4:71, Laube et al. (1993) JAMA 269: 2106-21-9; Elliott et al. (1987) Aust. Paediatr. J. 23: 293-297; Wigley et al. (1971) Diabetes 20: 552-556. Corthorpe et al. (1992) Pharm Res 9: 764-768; Govinda (1959) Indian J. Physiol. Pharmacol. 3: 161-167; Hastings et al. (1992) J. Appl. Physiol. 73: 1310-1316; Liu et al. (1993) JAMA 269: 2106-2109; Nagano et al. (1985) Jikeikai Med. J. 32: 503-506; Sakr (1992) Int. J. Phar. 86: 1-7; and Yoshida et al. (1987) Clin. Res. 35: 160-166 (each of which is referenced) Incorporated herein by reference). Pulmonary delivery of dry powder medication is described in US Pat. No. 5,254,330. A metered dose inhaler is described in Lee and Sciara (1976) J. Pharm. Sci. 65: 567-572. Intrabronchial administration of recombinant insulin is briefly described in Schlutiter et al. (Abstract) (1984) Diabetes 33: 75A and Kohler et al. (1987) Atemw. Lungenkrkr. 13: 230-232. Intranasal and pulmonary delivery of various polypeptides is described in US Pat. No. 5,011,678 and Nagai et al. (1984) J. Contr. Rel. 1: 15-22.

  Pharmaceutical compositions suitable for injectable use are known in the art and include, for example, sterile aqueous solutions (where water soluble) or suspensions and sterile solutions for the immediate preparation of sterile injectable solutions or dispersions. Contains powder. For intravenous administration of the ghrelin mimetic of this invention, suitable carriers include, for example, saline, bacteriostatic water, Cremophor EL ™ (BASF, New Jersey, Parsippany) or phosphate buffered saline (PBS ) Is included.

  The physiologically acceptable carrier can be any carrier known in the art as suitable for pharmaceutical (ie, topical, oral and parenteral) use. Suitable pharmaceutical carriers and formulations are described, for example, in Remington's Pharmaceutical Sciences (19th edition) (Genarro et al. (1995) Mack Publishing Co., Pennsylvania, Easton).

  Orally administered solutions generally comprise a physiologically acceptable inert diluent or edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For oral therapeutic administration, the ghrelin mimetics of this invention can be incorporated into physiological excipients and used in the form of tablets, troches or capsules.

  Several systems that alter the delivery of injectable drugs can be utilized to alter the pharmacokinetic properties of therapeutic agents (see, eg, K. Reddy, 2000, Annals of Pharmacotherapy 34: 915-923). . Drug delivery can be modified by changes in formulation (eg, continuous release products, liposomes) or addition to drug molecules (eg, PEGylation). Potential benefits of these drug delivery mechanisms include increased or prolonged duration of pharmacological activity, reduced adverse effects, and increased patient compliance and quality of life. Injectable continuous release systems are particularly suitable when it is important to deliver the drug in a controlled, predetermined manner and to avoid large fluctuations in drug concentration in plasma. Encapsulating a drug within a liposome can result in increased distribution to tissues (eg, tumors) with prolonged half-life and increased capillary permeability. PEGylation provides a method for modifying therapeutic peptides or proteins to minimize limitations (eg, stability, half-life, immunogenicity) that may be associated with the ghrelin mimics of this invention.

  According to this invention, at least one ghrelin mimetic is combined with a lipid or lipid vehicle (eg, micelles, liposomes, microspheres, protocells, protobionts, liposomes, coacervates, etc.) to allow multimer formation. Can do. Similarly, ghrelin mimetics can be multimerized using PEG addition, cross-linking, disulfide bond formation, covalent cross-linking formation, glycosyl phosphatidylinositol (GPI) anchor formation, or other established methods. it can. These multimerized ghrelin mimetics can be formulated into pharmaceutical compositions and utilized to increase or enhance their effectiveness.

  These ghrelin mimetics can also be manufactured in the form of suppositories (eg, using conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

  Those of skill in the art can utilize a variety of ghrelin mimetics of the present invention to more specifically deliver local gastrointestinal tissue such as, but not limited to, the stomach, esophagus, small intestine or colon. You will admit that the technology is known. This method of delivery will depend on factors such as the tissue of interest, the nature of the compound to be delivered, and the duration of treatment.

  In one aspect, the ghrelin mimetics of the present invention together with a carrier that protects the ghrelin mimetic against rapid elimination from the body, such as formulations for controlled release (including implants and microencapsulated delivery systems). Manufactured. These can be prepared according to methods known to those skilled in the art, for example, as described in US Pat. No. 4,522,811.

  Thus, although detailed preferred embodiments of the present invention have been described, the invention defined by the above paragraph is capable of many obvious equivalents without departing from the spirit or scope of the present invention. Therefore, it should be understood that it should not be limited to the specific ones described above.

  The invention will now be further illustrated by the following non-limiting examples.

  The gastrokinetic efficacy of ipamorelin was evaluated in a rat model of postoperative ileus. In these studies, ipamorelin is administered orally (10 mg / kg and 100 mg / kg) to male rats after abdominal surgery or as a single intravenous injection over a dosage range of 0.01 mg / kg to 1.0 mg / kg. Was administered. The effect of ipamorelin on gastric emptying was measured by administering ipamorelin immediately after administration of a dose of phenol red by oral gavage.

Example 1. Gastric motility efficacy of ipamorelin (10 or 100 mg / kg) in a rat model of postoperative ileus This study shows a single oral dose of ipamorelin at a dose of 10 or 100 mg / kg in a rat model of postoperative ileus Later, the potential gastric motility efficacy was assessed.

Methods and Experimental Design Male Sprague-Dawley CD ™ (Crl: CD ™ (SD)) rats (Rattus norvegicus) were obtained from Charles River Canada Inc. (St. Constant, Quebec, Canada). . In order to acclimate these animals to the laboratory environment, 9 days were allowed from the time they were obtained until the start of treatment. At the start of the treatment, the animals were about 7 weeks old and the body weight ranged from 230 to 254 g.

  The animals were individually housed in a stainless steel wire mesh bottom cage equipped with an automatic water supply valve. Each cage was clearly labeled with a color coded cage card indicating the project, group, number of animals and gender. Each animal was uniquely identified. The targeted conditions for the animal room environment and day length were as follows: temperature 22 ± 3 ° C .; humidity 50 ± 20%; photoperiod 12 hours of illumination and 12 hours of darkness.

  All animals had free access to a commercially available standard approved pelleted laboratory diet (PMI Certified Rodent Chow 5002: PMI Nutrition International Inc.), except during the indicated procedure. This diet was controlled and routinely analyzed by manufacturers for maximum allowable concentrations of contaminants (eg, heavy metals, aflatoxins, organophosphates, chlorinated hydrocarbons and PCBs). Municipal tap water that was softened, purified by reverse osmosis and exposed to ultraviolet light was given indefinitely (except during the indicated procedure). It appears that there were no known contaminants in the dietary material that could affect the results of this study.

  Ipamorelin obtained and used in this study was obtained from Bachem AG (Bubendorf, Switzerland). The vehicle used was 0.9% sodium chloride for injection (Baxter). The gastrointestinal marker used to assess gastric emptying levels was phenol red (Sigma Aldrich).

  An appropriate amount of the test article was dissolved in 0.9% sodium chloride for injection (US Pharmacopoeia). These test product formulations were adjusted to pH 7.4-7.5 using 0.1N / 1N hydrochloric acid or 0.1N sodium hydroxide as required. All dosing formulations were kept at room temperature and protected from light. Phenol red was prepared with deionized water as a 5 mg / mL solution on the day of dosing and stored protected from light at room temperature.

Surgical catheterization Each animal was injected with the antibiotics benzathine penicillin G and procaine penicillin G (0.1 mL) intramuscularly on the day of surgery and again on the second day after surgery. These animals were anesthetized with isoflurane gas and then prepared for surgery (including shaving of the thigh and dorsal incision sites). The shaved area was washed with chlorhexidine gluconate (4%) and then more povidone iodine (10%) was applied. Prior to surgery and at the end of the surgical procedure, a hypoallergenic ophthalmic lubricant (Tears Naturale ™ PM) was administered to each eye under anesthesia. Animals were maintained under isoflurane gas anesthesia throughout the surgical procedure.

  A small incision was made in the starboard area and the femoral vein was withdrawn. A small venous incision was made in the vein and a medical grade silicone-based catheter was inserted and the tip of the catheter was placed in the vena cava at approximately the kidney level. The catheter was secured in place with the appropriate suture material and passed through the skin to the exposed point of the neck of the neck. The thigh was cleaned by warm (about 37 ° C.) 0.9% sodium chloride injection (US Pharmacopeia). The thigh was closed with intermittent sew stitches and the exposed site was closed with purse string sutures (which were removed during 5-10 days or by healing results). Topical antibiotics (polymyxin B, bacitracin, neomycin) were administered daily until the end of this catheter exposure site and until deemed unnecessary at the thigh site.

  A jacket was placed over the animal to maintain a restraint system. A catheter prefilled with 0.9% sodium chloride injection (US Pharmacopeia) was sent through this restraint system and attached to a swivel secured to the outside of the cage. The top of the swivel was connected to an infusion pump and all animals were continuously infused with 0.9% sodium chloride injection (US Pharmacopeia) at a rate of 0.4 mL / hour until the start of treatment.

All treatment procedures for inducing post-operative ileus were repeated over two consecutive days with approximately equal numbers of animals from each group treated each day. No food was given to the animals overnight before surgery. On the day of surgery, the animals were anesthetized with isoflurane gas and a non-irritating ophthalmic lubricant (Tears Naturale PM) was applied to each eye. These animals were prepared for surgery by shaving the entire abdominal area. The shaved area was cleaned and properly sterilized before incision. Using a scalpel, the abdomen was opened to locate the cecum. The cecum was removed from the body and treated for about 1 minute (ie, gently tapped between both hands in a gauze soaked with saline). The cecum was then returned to its original position and the surgical site was closed with absorbable suture material (intermittent suture) and staples. The animals were then returned to their cages to recover from anesthesia.

  The animals assigned to the sham control group (Group 1) received the same treatment procedure, except that the cecum was removed from the body and returned to its original position without treatment.

Administration dosage of the dose begins daily continuous, approximately equal numbers of animals from each group were dosed on each day. Prior to dosing, these animals were taken up with water. These test / control articles were administered by oral gavage using a syringe and a flexible gavage tube. These animals were dosed immediately following administration of phenol red by oral gavage. The dose volume was 5 mL / kg (for both ipamorelin and phenol red doses) and the actual dose administered was based on the immediate body weight of each animal.

Gastrointestinal assessment Approximately 30 minutes after morphine injection, all animals were given 0.4 mL of phenol red by oral gavage. Approximately 30 minutes after administration of the control or test article, these rats were euthanized. The stomach was then revealed by abdominal wall incision and quickly ligated at the pylorus and cardia and removed. The stomach was cut open and the contents extracted with 100 mL 0.1 N NaOH. The phenol red content of this extract was assayed spectrophotometrically at 558 nm on a spectrometer. After collection, the samples were stored on wet ice until moved for analysis.

Results Ipamorelin administered orally after postoperative ileus accelerated gastric emptying by 12.4% and 41.6%, respectively, compared to control animals at 10 and 100 mg / kg (provided that Not achieved statistical significance). See FIG.

Conclusion Ipamorelin administered orally at doses of 10 or 100 mg / kg accelerated gastric emptying in a rat model of postoperative ileus in a dose-dependent relationship.

Example 2 Gastric motility efficacy of intravenously administered ipamorelin (0.1, 0.25 or 1.0 mg / kg) in a rat model of postoperative ileus In this study, ipamorelin was administered by slow intravenous bolus injection via an indwelling catheter (In approximately 100 seconds).

Methods and Experimental Design Male Sprague-Dawley CD ™ (Crl: CD ™ (SD)) rats (Rattus norvegicus) were purchased from Charles River Canada Inc. (St. Constant, Quebec, Canada). . There was an 8 day period from the purchase of these animals to the surgical implantation of the catheters, and these animals were adapted to physical and environmental conditions. Animal dosing began approximately one week after surgical implantation of the catheter to allow proper recovery of the animal prior to treatment. At the start of the treatment, the animals were about 10-12 weeks old and the body weight ranged from 327-397 g.

  The animals were individually housed in a stainless steel wire mesh bottom cage equipped with an automatic water supply valve. Each cage was clearly labeled with a color coded cage card indicating the project, group, number of animals and sex. Each animal was individually identified. The targeted conditions for the animal room environment and day length were as follows: temperature 22 ± 3 ° C .; humidity 50 ± 20%; photoperiod was 12 hours of illumination and 12 hours of darkness .

  All animals had free access to standard commercial approved pelletized laboratory food (PMI CertifiedRodent Chow 5002: PMI Nutrition International Inc.). This diet was controlled and routinely analyzed by the manufacturer for maximum allowable concentrations of contaminants (eg, heavy metals, aflatoxins, organophosphates, chlorinated hydrocarbons and PCBs). Unlimited municipal tap water that was softened, purified by reverse osmosis and exposed to UV light (except during the indicated procedure). There appears to be no known contaminants in these dietary materials that affect the results of this study.

  Ipamorelin, obtained and used in this study, was obtained from Bachem AG. The vehicle used was 0.9% sodium chloride (Baxter) for injection. The gastrointestinal marker used to assess the level of elimination from the stomach was phenol red (Sigma Aldrich).

Surgical Catheterization Each animal received an intramuscular injection of the antibiotics benzathine penicillin G and procaine penicillin G (0.1 mL) on the day of surgery and again on the second day after surgery. These animals were anesthetized with isoflurane gas and then prepared for surgery such as shaving of the thigh and dorsal incision sites. The shaved area was washed with 4% chlorhexidine gluconate and then with more 10% povidone iodine. Prior to surgery and at the end of the surgical procedure, a hypoallergenic lubricating ophthalmic drug (Tears Naturale PM) was administered to each eye under anesthesia. Animals were maintained under isoflurane gas anesthesia throughout the surgical procedure.

  A small incision was opened in the starboard area and the femoral vein was withdrawn. A small venous incision was made and a medical grade silicone-based catheter was inserted. The catheter tip was placed in the vena cava at approximately the kidney level. The catheter was secured in place with sutures and then guided subcutaneously to the exposed point in the neck of the neck. The thighs were washed with warm (about 37 ° C.) 0.9% sodium chloride injection (US Pharmacopoeia). The thigh was closed with intermittent seam stitches, and the exposed area was closed with purse string sutures, and the thread was removed after 5-10 days or depending on the healing results. Topical antibiotics (polymyxin B, bacitracin, neomycin) were administered daily to the catheter exposure site to the end and to the thigh until deemed unnecessary.

  A jacket was placed over the animal to maintain a restraint system. A catheter prefilled with 0.9% sodium chloride injection (US Pharmacopeia) was fed through this restraint system and attached to a swivel secured to the outside of the cage. The top of the swivel was connected to an infusion pump and all animals were continuously infused with 0.9% sodium chloride injection (US Pharmacopeia) at a rate of 0.4 mL / hour until the start of treatment.

All treatment procedures for inducing post-operative ileus were repeated over two consecutive days with approximately equal numbers of animals from each group treated each day. No food was given to the animals overnight before surgery. On the day of surgery, the animals were anesthetized with isoflurane gas and a non-irritating ophthalmic lubricant (Tears Naturale PM) was applied to each eye. These animals were prepared for surgery by shaving the entire abdominal area. The shaved area was cleaned and properly sterilized before incision. Using a scalpel, the abdomen was opened to locate the cecum. The cecum was removed from the body and treated for about 1 minute (ie, gently tapped between both hands in a gauze soaked with saline). The cecum was then returned to its original position and the surgical site was closed with absorbable suture material (intermittent suture) and staples. The animals were then returned to their cages to recover from anesthesia.

  Animals assigned to the fake control group (Group 1) received the same treatment procedure, except that the cecum was removed from the body and returned to its original position without treatment. Due to technical carelessness, animal no. Note that the cecum of 2001, 2009, 3001 and 4001 were treated with sterile water for injection (US Pharmacopoeia) instead of gauze infiltrated with saline solution. This slight deviation was considered to have no effect on the results of this study or the interpretation of the results.

Administration dosage of the dose begins daily continuous, approximately equal numbers of animals from each group were dosed on each day. These test / control articles were administered as slow bolus injections (approximately 100 seconds) via an indwelling catheter. These animals were administered immediately after administration of phenol red by oral gavage. The dose volume was 5 mL / kg (both ipamorelin and phenol red administration), and the actual dose administered was based on the immediate body weight of each animal.

Gastrointestinal Evaluation Prior to administration of phenol red, these animals were taken up with water. Approximately 30 minutes after surgery (ileus), the animals received 0.4 mL of phenol red by oral gavage. The animals were then dosed and euthanized approximately 30 minutes later. At euthanasia, the stomach was exposed by abdominal wall incision and quickly ligated and removed at the pylorus and cardia. The stomach was cut open and the contents extracted with 100 mL 0.1 N NaOH. The phenol red content of this extract was assayed spectrophotometrically at 558 nm by a colorimetric method. After collection, the samples were stored on wet ice until moved for analysis.

Results After surgery, ipamorelin, administered intravenously at doses of 0.1, 0.25 and 1.0 mg / kg, caused gastric emptying in male albino rats with vehicle and fake control animals. Accelerated compared (although no dose relationship was observed). Animals treated with intravenous dosing of ipamorelin (0.1, 0.25, or 1.0 mg / kg) reduced gastric phenol red content compared to the vehicle control group (85.7 respectively). %, 95.6% and 92.2%). These reductions reached statistical significance at the 0.25 and 1.0 mg / kg dose levels (p ≦ 0.001 and p ≦ 0.01, respectively). See FIG.

  The average gastric phenol red content of the vehicle control group is similar to that of the fake control group and is not statistically different, which is detected in the (unexpected) ileus or surgical control group in the fake control group It may reflect the inability to induce possible ileus. Therefore, animals treated with intravenous dosing of ipamorelin (0.1, 0.25 or 1.0 mg / kg) when expressed relative to the fake control group have reduced gastric phenol red content (87.1%, 96.0% and 93.1%, respectively), which were statistically significant at all dose levels. See FIG.

Conclusions Ipamorelin administered intravenously to male albino rats at doses of 0.1, 0.25 and 1.0 mg / kg after surgery compared gastric emptying compared to sham and vehicle control animals. Significantly accelerated.

Example 3 Gastric motility efficacy of intravenously administered ipamorelin (0.01, 0.03 or 0.1 mg / kg) in a rat model of postoperative ileus In this study, ipamorelin was administered as an indwelling catheter as a slow bolus intravenous injection. Via a dose of 0.01, 0.03 or 0.1 mg / kg (approximately 100 seconds).

Male Sprague-Dawley CD ™ (Crl: CD ™ (SD)) rats (Rattus norvegicus) were used. Seven days from purchase of these animals to surgical implantation of the catheters allowed them to acclimate to physical and environmental conditions. Dosing to these animals began approximately 1 week after surgical implantation of the catheter for proper recovery prior to animal treatment. At the start of treatment, the animals were approximately 10 weeks old and the body weight ranged from 334 to 385 g. The animals were individually housed in a stainless steel wire mesh bottom cage equipped with an automatic water supply valve. Targeted conditions for animal room environment and day length were as follows:
Temperature: 22 ± 3 ° C .; Humidity: 50 ± 20%; Photoperiod: 12 hours of illumination and 12 hours of darkness.
All animals were examined twice daily for mortality and signs of illness and response to treatment (provided that the day of arrival and the date of necropsy are examined once). Individual body weights were measured randomly the day before dosing (for dose calculation purposes only).

Surgical catheterization A small incision was made in the starboard region and the femoral vein was withdrawn. A small venous incision was made and a medical grade silicone-based catheter was inserted and the tip of the catheter was positioned in the vena cava at approximately the kidney level. The catheter was secured in place with an appropriate suture and then passed subcutaneously to the exposed point of the neck nape. The thighs were washed with warm (about 37 ° C.) 0.9% sodium chloride injection (US Pharmacopoeia). The thigh was closed using intermittent sew stitches, the exposed site was closed with purse string sutures, and the thread was removed within 5-10 days or by healing results. Topical antibiotics (polymyxin B, bacitracin, neomycin) were administered to the catheter exposure site to the end of each day and to the thigh until deemed unnecessary.

  A jacket was placed over the animal to maintain a restraint system. A catheter prefilled with 0.9% sodium chloride injection (US Pharmacopeia) was fed through this restraint system and attached to a swivel secured to the outside of the cage. The top of the swivel was connected to an infusion pump and all animals were continuously infused with 0.9% sodium chloride injection (US Pharmacopeia) at a rate of 0.4 mL / hour until the start of treatment.

The entire surgical procedure to induce post-operative ileus was repeated for 2 consecutive days with approximately equal numbers of animals from each group treated each day. No food was given to the animals overnight before surgery. On the day of surgery, animals were anesthetized with isoflurane gas and a mild eye lubricant (Tears Naturale PM) was applied to each eye. These animals were prepared for surgery by shaving the entire abdominal area. The shaved area was cleaned and properly sterilized before incision. Using a scalpel, the abdomen was opened to locate the cecum. The cecum was removed from the body and treated for about 1 minute (ie, gently tapped between both hands in a gauze soaked with saline). The cecum was then returned to its original position and the surgical site was closed with absorbable suture material (intermittent suture) and staples. The animals were then returned to their cages to recover from anesthesia.

  Animals assigned to the non-operative control group (Group 1) did not undergo surgery to induce postoperative ileus.

Administration dosage of the dose begins daily continuous, approximately equal numbers of animals from each group were dosed on each day. These test / control articles were administered as a slow bolus intravenous injection via an indwelling catheter. These animals were dosed immediately after administration of phenol red by oral gavage. The dose volume was 5 mL / kg for groups 1, 2 and 5; 0.5 mL / kg for group 3 and 1.5 mL / kg for group 4. The actual dose administered was based on the most recent body weight of each animal.

Gastrointestinal Evaluation Prior to administration of phenol red, these animals were taken up with water. Approximately 30 minutes after surgery (ileus), the animals received 0.4 mL of phenol red by oral gavage. The animals were then dosed and euthanized approximately 30 minutes later. At euthanasia, the stomach was exposed by abdominal wall incision and quickly ligated and removed at the pylorus and cardia. The stomach was cut open and the contents extracted with 100 mL 0.1 N NaOH. The phenol red content of this extract was assayed spectrophotometrically at 558 nm by a colorimetric method. After collection, the samples were stored on wet ice until moved for analysis.

Results After surgery, ipamorelin administered intravenously at doses of 0.01, 0.03 and 0.1 mg / kg reduced gastric emptying in male albino rats, vehicle control and non-operative control animals. (Dose relationship was not observed). Animals treated with intravenous dosing of ipamorelin (0.01, 0.03 or 0.1 mg / kg) showed a reduction in stomach phenol red content compared to the vehicle control group. These reductions reached statistical significance at all dose levels (0.01 and 0.1 mg / kg, p ≦ 0.05 and 0.03 mg / kg, p ≦ 0.0). See FIG.

  The average gastric phenol red content of the vehicle control group is similar to that of the non-operative control group and is not statistically different, reflecting the inability to induce detectable ileus in the surgical control group. sell. Thus, when expressed relative to the non-surgical control group, animals treated with intravenous ipamorelin (0.01, 0.03 or 0.1 mg / kg) were treated with gastric phenol red content. There was a decrease, which was statistically significant at all dose levels.

Example 4 Efficacy of Morphine Inducing Gastric Ileus in Rats The purpose of this study was to evaluate the potential efficacy of inducing gastric ileus in morphine rats. The treatment groups are shown below.

Methods and Experimental Design Male Sprague-Dawley CD ™ (Crl: CD ™ (SD)) rats (Rattus norvegicus) were obtained from Charles River Canada Inc. (St. Constant, Quebec, Canada). . Upon arrival, all animals were subjected to a general physical examination by qualified members of the veterinary staff to ensure normal health. All animals were considered acceptable for use. Nine days were allowed to acclimate the animal to the laboratory environment from the time the animal was received until the start of treatment. At the start of treatment, the animals were approximately 7 weeks old and the body weight ranged from 231 to 267 g. Note that the animal's weight was slightly outside the range stated in the protocol. This bias was considered to have no impact on the results of this study or the interpretation of the results.

  The animals were individually housed in stainless steel wire mesh bottom cages with automatic water supply valves. The targeted conditions for the animal room environment and day length were as follows: temperature 22 ± 3 ° C .; humidity 50 ± 20%; photoperiod 12 hours of illumination and 12 hours of darkness.

  All animals had free access to commercially available standard approved pelletized laboratory food (PMI Certifie Rodent Chow 5002: PMI Nutrition International Inc.) except in the indicated procedure. This diet was controlled and routinely analyzed by manufacturers for maximum allowable concentrations of contaminants (eg, heavy metals, aflatoxins, organophosphates, chlorinated hydrocarbons and PCBs). The results of these analyzes are retained in the PCS-MTL scientific archive. Municipal tap water that was softened, purified by reverse osmosis and exposed to ultraviolet light was given indefinitely (except during the indicated procedure). Periodic analysis of water was subcontracted to a management accredited analytical laboratory audited by the quality assurance department of PCS-MTL. The results of these analyzes are retained in the PCS-MTL scientific archive. It appears that there were no known contaminants in the dietary material that could affect the results of this study.

  These dosage formulations were prepared on the day of administration. Appropriate controls were dissolved in the vehicle to achieve the desired concentration. A morphine sulfate solution was used as the feed. Phenol red was prepared as a 5 mg / mL solution on the day of administration and stored at room temperature protected from light until use.

Dosage doses of morphine (4 mg / kg or 1 mg / kg) were administered by subcutaneous injection using a hypodermic needle attached to a syringe on the upper back (scapular region). Morphine was administered approximately 30 minutes before administration of the control / positive control product. The dose volume was 0.1 mL / kg for groups 2 and 3, and 0.4 mL / kg for groups 4 and 5. The actual dose administered was based on the most recent body weight of each animal.

Gastrointestinal assessment Approximately 30 minutes after morphine injection, all animals were given 0.4 mL of phenol red by oral gavage. Approximately 30 minutes after administration of the control or test article, these rats were euthanized. The stomach was then revealed by abdominal wall incision and quickly ligated at the pylorus and cardia and removed. The stomach was cut open and the contents extracted with 100 mL 0.1 N NaOH. The phenol red content of this extract was assayed spectrophotometrically at 558 nm on a spectrometer. After collection, the samples were stored on wet ice until moved for analysis.

Results When administered subcutaneously at a dose of 1 or 4 mg / kg, morphine significantly reduces gastric emptying by approximately 42% compared to control animals (but does not achieve statistical significance). Ghrelin, a potent gastric motility peptide known to reverse gastric ileus, reduces gastric emptying when administered intravenously at a dose of 50 μg / kg after exposure to 1 mg / kg morphine. Accelerated by approximately 37% and reduced ileus by approximately 11% when compared to animals treated with 1 mg / kg and saline solution. However, ghrelin did not accelerate gastric emptying in rats dosed with 4 mg / kg morphine, and both groups (saline solution vs. ghrelin) showed the same group mean absorbance of phenol red (respectively, 2.99 and 3.01). See FIG.

CONCLUSION Morphine administered subcutaneously to male albino rats at doses of 1 or 4 mg / kg induced similar levels of gastric ileus in a non-dose-dependent manner. Ghrelin accelerated gastric emptying and reversed gastric ileus induced after exposure to 1 mg / kg morphine, but gastric emptying in mice receiving a 4 mg / kg dose of morphine Had no effect.

Example 5 FIG. Intravenous injection of ipamorelin (1.0, 2.5, 10 mg / kg) to promote gastric motility compared to RC-1139 to treat postoperative ileus in a rat model In an ileus rat model, it was evaluated in comparison with RC-1139 (a ghrelin mimetic with known gastric motility enhancing potency). In this study, each drug was administered as a single intravenous injection as shown in the table below. The treatment groups were as follows:

  All animals were examined twice daily for mortality and signs of unhealthy or response to treatment (except on the day of arrival when the animals were tested once and the date of necropsy). Individual body weights were measured randomly and on the day before dosing (for purposes of dose calculation only). Since the two animals died immediately after administration of ipamorelin at a dose of 10 mg / kg, all remaining animals in group 4 were dosed at 0.25 mg / kg.

Methods and Experimental Design Male Sprague-Dawley CD ™ (Crl: CD ™ (SD)) rats (Rattus norvegicus) were randomly divided into multiple treatment groups. There was at least 5 days between receiving the animal and starting treatment to allow the animal to acclimate to the laboratory environment. At the start of treatment, the animals were about 7 weeks old and weighed in the range of 205-272 g.

  Prior to the preparation of the formulation for the first administration, a pilot preparation was made at 2 mg / mL (test article solution) and 2 mg / mL (reference article solution) to confirm the suitability of the proposed formulation.

  For administration, formulations were prepared by dissolving an appropriate amount of test or reference product in 0.9% sodium chloride for injection (US Pharmacopeia). These dosage formulations were then adjusted to pH 7.4-7.5 using 0.1N / 1N hydrochloric acid or, if necessary, 0.1N sodium hydroxide. All dosage formulations were filtered through a 0.2 μm PVDF filter before use and stored at room temperature protected from light.

  Phenol red was prepared in deionized water as a 5 mg / mL solution on the day of administration and stored at room temperature protected from light.

All surgical procedures to induce post-operative ileus were repeated for two consecutive days with approximately the same number of animals from each group treated each day. No food was given to the animals overnight before surgery. On the day of surgery, animals were anesthetized with isoflurane gas and a mild eye lubricant was applied to each eye. These animals were prepared for surgery by shaving the entire abdominal area. The shaved area was cleaned and properly sterilized before incision. Using a scalpel, the abdomen was opened to locate the cecum. The cecum was removed from the body and treated for about 1 minute (ie, gently tapped between both hands in a gauze soaked with saline). The cecum was then returned to its original position and the surgical site was closed with absorbable suture material (intermittent suture) and staples. The animals were then returned to their cages to recover from anesthesia.

Dosage dose animals were dosed immediately after administration of phenol red by oral gavage. These test / control and reference products were administered by intravenous injection (slow bolus injection for approximately 100 seconds) into the tail vein using a syringe and an appropriate gauge needle. The dose volume was 5 mL / kg and the actual dose administered was based on the immediate body weight of each animal.

Gastrointestinal evaluation Prior to administration of phenol red, it was noted that these animals were not given water. Approximately 30 minutes after surgery, the animals were given 0.4 mL of phenol red by oral gavage followed by a test article and then euthanized approximately 30 minutes later. At euthanasia, the stomach was exposed by abdominal wall incision and quickly ligated and removed at the pylorus and cardia. The stomach was cut open and the contents extracted with 100 mL 0.1 N NaOH. The phenol red content of this extract was assayed spectrophotometrically at 558 nm by a colorimetric method. After collection, the samples were stored on wet ice until moved for analysis.

Results After induction of ileus, ipamorelin administered intravenously at doses of 0.25, 1 and 2.5 mg / kg reduced gastric emptying, in male albino rats and in control animals and RC-1139. Accelerated compared to treated animals (although no dose relationship was observed). See FIG.

  The 0.25 and 2.5 mg / kg ipamorelin doses show similar levels of potency of about 50 and 60% reduction in phenol red content, respectively, and about 21% after 1 mg / kg dose. Gastric emptying was observed. The stomach content of phenol red marker after administration of 10 mg / kg RC-1139 was about 16% lower than that of the control group.

Conclusions Ipamorelin, administered intravenously at doses of 0.25, 1, and 2.5 mg / kg to male albino rats with postoperative ileus, accelerates gastric emptying compared to control and reference animals did.

Example 6 Administration of ipamorelin to reverse morphine-induced gastric emptying delay in healthy volunteers
Delayed introduction of gastric contents plays an etiologic role in several important target indications including postoperative ileus, opiate-induced colorectal dysfunction and gastric failure paralysis. In addition, delayed gastric emptying can promote or worsen nausea. Thus, drugs such as ipamorelin have the demonstrated ability to promote gastric emptying in animal models (see examples above) and are important therapeutic agents in various GI diseases classified by reduced motility Can play a role.

  Ipamorelin is a ghrelin mimetic. Ghrelin is a 28 amino acid peptide hormone that is synthesized primarily in the gastric acid secretory gland and, to a lesser extent, in other organs of the body such as the kidney and hypothalamus. Among the important physiological effects exerted by ghrelin is the ability to regulate gastric motility, which has shown strong gastric motility promoting effects in various animal species and humans (upper and lower GI). In both). See Masuda 2000, Asakawa 2001, Tack 2006. In addition, in rat models, ghrelin has been shown to eliminate gastric postoperative ileus. See Trudel 2002.

  It is likely that ghrelin's gastric motility promoting activity is mediated by direct effects on the gastrointestinal tract or indirectly by the vagal cholinergic muscarinic pathway. It acts locally in the stomach and stimulates the movement of the stomach by stimulating the firing of vagal afferent neurons. See Peeters 2003. Efforts have been made over the years to exploit the positive effects of ghrelin in various diseases by identifying and developing drugs that mimic ghrelin. Ghrelin has an exceptionally short half-life (about 10 minutes) in humans and therefore has a limited therapeutic potential. Ipamorelin has a half-life of about 6 hours available as an intravenous therapeutic agent in humans and is therefore a ghrelin mimetic suitable for therapeutic applications.

  This study was designed to employ a well-proven clinical pharmacological model (acetaminophen AUC) for assessing the effect of ipamorelin on gastric emptying. Ipamorelin was shown to have a strong stimulatory effect on gastric emptying in the rat model in the above examples. This study was intended to extend these findings to humans.

Dosage The dose of ipamorelin selected for this study (0.06 mg / kg intravenous infusion over 15 minutes) has been shown to be safe and well tolerated in previous Phase 1 studies. Dose. Ipamorelin was formulated as a 0.5 mg / mL sterile solution using 2 equivalents of acetic acid in normal saline solution. This sterile solution was further diluted with normal saline solution to a volume suitable for administration before use.

  The dose of morphine selected for this study (0.05 mg / kg, intravenous bolus administration) is clinically relevant as an analgesic dose, and significantly reduces gastric emptying in normal volunteers. It was the [Yuan 1998] dose previously shown to be delayed.

  The dose of acetaminophen selected for this study (1000 mg oral suspension dose) is a standard acetaminophen drug dose. This dose has been successfully adopted in previous gastric emptying studies, resulting in an easily measured plasma concentration (> 0.2 μg / mL).

The study design was a standard, three-phase, randomized, single-dose crossover study.
This study is performed using a single dose administration, which is well-studied and suitable for the acetaminophen AUC model of gastric emptying. All of the three drugs employed in this study have a half-life of 6 hours or less, and therefore a wash interval of 5-8 days ensures complete elimination of these drugs from the body. .

The purpose of this study was to evaluate the ability of intravenous ipamorelin to reverse the delay in opiate-induced gastric emptying and to evaluate the ability of intravenous ipamorelin to reverse opiate-induced nausea. Inclusive. We reverse the gastric emptying delay induced by opaate for ipamorelin as assessed by plasma acetaminophen absorption (AUC _0-60 after ipamorelin administration is 50% higher than that after placebo administration) Predicted).

Study Design This study was conducted as a single-centered, double-blind, randomized, single-dose, three-way crossover study. This study compared the following treatments:
1. Morphine + ipamorelin;
2. 2. Morphine control; Normal control

As a measure of gastric emptying, plasma samples were obtained over 3 hours after administration of acetaminophen for the measurement of acetaminophen AUC. It is expected that morphine will significantly delay gastric emptying [Yuan 1998]; in addition, it is expected that ipamorelin will reverse the observed delay of gastric emptying. It was. The first parameter of interest is the initial absorption of acetaminophen as reflected in the plasma AUC over the first hour (AUC _0-60 ) after administration of acetaminophen. Further parameters of interest include AUC _0-180 , C _MAX and T _MAX .

  The drug for this study was administered over a 15 minute period via a well-calibrated infusion pump (eg, Harvard pump or similar). The dose for each patient was calculated up to 100 kg (6 mg) based on body weight. The dosing volume was then diluted to a total volume of 15 mL using normal injectable salt solution as a diluent. Bubbles were drawn into the syringe (to facilitate agitation) and mixed by gently inverting the syringe 6 times.

Morphine administration Morphine (1.0 mg / mL) / placebo was administered by slow bolus injection (30-60 seconds). The infusion catheter was then immediately flushed with 3-5 mL normal saline solution.

Acetaminophen Administration Acetaminophen suspension (32 mg / mL) was shaken well before administration. The dose to be administered was 31 mL (992 mg). The patient then drank given an additional 150 mL of water.

Results Treatment with ipamorelin was well tolerated and these results showed that morphine-induced delays in gastrointestinal motility were reversed in humans. See FIG.

Example 7 A lower dose study of ipamorelin in delayed morphine-induced gastric emptying in healthy male volunteers. This study was the same design as Example 6a, but induced by an opiate of gastric emptying The lower intravenous dose of ipamorelin that reverses the delayed effect was evaluated. Data are given for 23 patients who completed all treatments. The process of this study is:
(1) untreated (saline solution) control; (2) morphine 0.05 mg / kg (IV: intravenous administration); and (3) morphine 0.05 mg / kg + ipamorelin 0.01 mg / kg (iv administration) and (4) Morphine 0.05 mg / kg + ipamorelin 0.03 mg / kg (intravenous administration). Acetaminophen elixir was administered orally in each treatment cycle to allow assessment of gastric content exclusion. These treatments were given in a single-blind, placebo-controlled, three-way crossover study (with 5-8 days of washing between treatments).

  These results indicate that morphine administration delayed gastric emptying as measured by a decrease in plasma acetaminophen levels. This effect was reversed by the dose of ipamorelin and was comparable to that presented in Example 6. The data is shown in FIG.

Example 8 FIG. Comparison of effects of various ghrelin mimetics, including ipamorelin, on gastrointestinal motility in rats The purpose of this study was to determine the pharmacological effects of a series of ghrelin mimetics on gastrointestinal motility (measured by passage of charcoal) in rats in general. The gastrointestinal motility enhancer used was metoclopramide and was evaluated after a single intravenous infusion of the experimental drug compared to the control. The treatment groups were as follows:

Procedure:
Each dosing formulation was prepared on the day of dosing. For these ghrelin mimetics, an aliquot of a 4 mg / mL stock solution was diluted with an appropriate volume of vehicle to achieve the final desired concentration. A reference formulation was also prepared on the day of administration by mixing an appropriate amount of reference with an appropriate amount of vehicle to achieve the desired final concentration. The supplied salt solution and metoclopramide (1 mg / mL) were used.

  Dosing was performed on consecutive days, and approximately the same number of animals were dosed each day. No food was given to these animals overnight before surgery. Each formulation was administered by intravenous injection into the tail vein using a syringe and an appropriate gauge needle. The dose volume was 5 mL / kg or 10 mL / kg (for metoclopramide only).

  Approximately 30 minutes after administration of the test article, all animals received 3 mL of activated charcoal by oral gavage and no food or water was given to the animals for the next 20 minutes.

  After the observation period, these rats were euthanized, the abdominal cavity was opened, and the stomach and small intestine were removed. Recorded the presence or absence of charcoal. The weight of these stomachs (with or without contents) was measured to provide an index of gastric emptying. These small intestines were opened and expanded over their entire length. The position of the charcoal was located and the distance from the pyloric sphincter to the closest and far away charcoal and the total distance from the pyloric sphincter to the cecum was measured. The distance traveled by the charcoal was also measured as a percentage of the total length of the small intestine.

Results Metoclopramide significantly increased small intestinal motility and increased the distance traveled by the charcoal diet by 29%, from 67.3% in the control group to 86.9% in the metoclopramide treated group. In contrast, the gastric charcoal content of the group of animals receiving metoclopramide was not significantly different from that of the control group.

  Ipamorelin significantly increased gastric emptying compared to the control group, reducing the amount of charcoal remaining in the stomach by 66%. Ghrelin and GHRP-6 also significantly reduced the amount of charcoal remaining in the stomach compared to the control group (57% and 64%, respectively), but their effects were not significantly different from that of ipamorelin. It was.

  Ipamorelin apparently produced a very significant (P ≦ 0.001) increase in gastric emptying and a trend towards increased small intestinal motility compared to the saline treated control group, Supports the view that ipamorelin is a potent gastrointestinal motility promoter. The results of these experiments are shown in FIGS.

Example 9 Comparison of effects of various ghrelin mimetics including ipamorelin on gastrointestinal motility in rats The purpose of this study was to determine the pharmacological effects of a series of ghrelin mimetics on gastrointestinal motility in rats (by measuring charcoal transit) It was to evaluate after a single intravenous infusion compared to controls.

Procedure:
Dosing was started on consecutive days and approximately the same number of animals from each group were dosed each day. No food was given to these animals overnight before surgery. Prior to administration, these animals were not given water. These test / reference / positive controls were administered by intravenous injection into the tail vein using a syringe and appropriate gauge needle. The dose volume was 5 mL / kg and the actual dose administered was based on the immediate body weight of each animal.

  Approximately 30 minutes after administration of the test / reference or positive control product, all animals received 3 mL of activated charcoal by oral gavage, and then no food and water were given to the animals for 20 minutes.

  At the end of this observation period, the rats were euthanized, the abdominal cavity was opened, and the stomach and small intestine were removed. The presence or absence of charcoal in the stomach was recorded. These stomachs (with and without contents) were weighed and recorded to obtain an index of gastric emptying. These small intestines were opened and expanded over their entire length. The position of the charcoal was located and the distance from the pyloric sphincter to the nearest and farthest charcoal traces and the total distance from the pyloric sphincter to the cecum were measured and recorded (all distances were measured in mm). In addition to stomach weight (with and without content), the distance traveled by charcoal was recorded as a percentage of the total length of the small intestine to obtain an index of gastric emptying. Any abnormal or unusual clinical signs observed during this 20 minute observation period were recorded.

Results Ghrelin and the three ghrelin mimetics tested in this study (ipamorelin, GHRP-6 and RC-1139 acetate) were administered independently in male albino rats at a single intravenous dose of 75 μmol / kg. In some cases, it showed different effects on gastric emptying and small intestine passage. Only ipamorelin showed a similar stimulatory effect to ghrelin on the measurement of gastric emptying and passage through the small intestine (the movement of charcoal pyloric sphincter at short and long distance). Ghrelin, GHRP-6, and ipamorelin caused gastric emptying by one of two measurements of small intestine passage (short distance movement of charcoal from the pyloric sphincter). See FIGS.

Example 10 Efficacy of ipamorelin for the treatment of postoperative ileus in rats The purpose of this study was to investigate whether ipamorelin accelerates colonic transit in a rat model of POI. To make this assessment, both single dose and multiple dose efficacy were evaluated.

Materials and Methods Animals: Adult male Sprague-Dawley rats with indwelling catheters implanted in the right jugular vein were obtained from Charles River (Massachusetts, Wilmington). The initial body weight of these animals was 250-270 g. These catheters were maintained patently during environmental acclimatization and were utilized for ipamorelin or vehicle dosing. A further group of control rats that are not subjected to surgery and are not subjected to drug or vehicle treatment, indwelling catheters used to instill dye markers that measure colonic passage are implanted in the proximal colon (1-2 cm from the cecum) Purchased a group that has been. All rats were housed alone under controlled conditions (25 ° C., 12 hour light / dark cycle) and had free access to food and water. An environmental acclimation period of at least one week was given prior to testing.

  Induction of post-operative ileus (POI): POI was induced by the surgical procedure described in “running of the bowel” (Kalff et al., 1998). In particular, rats were anesthetized by inhalation of isoflurane (2-3%), the abdomen was shaved, sterilized, and a midline incision was made to expose the viscera. The small intestine and cecum were probed using a cotton applicator that was pulled out of the body and soaked in sterile saline for 5 minutes. After completion of the review, the intestine was covered with gauze soaked in saline solution and the abdomen was kept open for an additional 10 minutes. To study colon transit, 200 μl of non-absorbable dye marker (trypan blue in saline solution) was injected into the proximal (1 cm distance from the cecum) colon. The incision was then closed using silk sutures. The whole procedure lasted 25-30 minutes. Surgery was always performed from 6:00 am to 8:00 am, and these animals received ipamorelin or vehicle treatment during light-dark cycle illumination.

  Measurement of colon transit time: Prior to the experiment, these rats were fasted for 20-22 hours, with free access to water. At the end of the surgical procedure, these rats received an intracolonic injection of the dye marker. After this surgery, the rats were placed in a clean cage and fed a premeasured amount of food (Purina ™ rat chow) and water. Colon transit time was evaluated as the period between the end of surgery and the appearance of dye in the feces. Inexperienced rats that had not undergone surgery and treatment with drugs or vehicle were studied on each experimental day with rats with POI. These naive rats were fitted with a colon catheter that was used to instill a dye marker into the colon after 20-22 hours of fasting. Data collected from these animals served as a reference to healthy controls. In previous studies, rats with POI showed a significant delay in colon transit time compared to naive animals (Zittel et al., 1998; Greenwood-Van Meerveld, unpublished data).

  Cumulative fecal excretion, food intake and body weight: Feces were counted and weighed at 3 and 12 hour intervals during the first 48 hours after surgery (see experimental design). This cumulative fecal excretion was assessed by adding the number of fecal particles during 48 hours after surgery. Dietary intake was recorded at 3 or 12 hour intervals according to the experimental design and normalized as g per 100 g body weight. Cumulative food intake was calculated over 48 hours after surgery in both series of experiments. Body weights were measured daily from 8:00 am to 9:00 am before animals were fasted and on the day of the experiment they were measured before surgery and 24 and 48 hours after surgery. The change in body weight was expressed as weight gain compared to the weight of fasted animals measured before surgery.

Test and Control Supplies: The test compound ipamorelin (free base) was converted to the diacetate by mixing with 2 molar equivalents of glacial acetic acid. A 0.5 mg / ml stock solution was prepared daily with sterile saline solution and glacial acetic acid (0.1 μl / ml) to make ipamorelin as a solution (pH 3-4). The solution was then titrated with NaOH to pH 7.0-7.2. Further dilution was performed with saline solution. Sterile saline solution was used in vehicle control experiments. The positive control [D-Lys ³ ] -GHRP-6 was purchased from Sigma-Aldrich (Missouri, St. Louis) and dissolved in this saline solution. Both test and control articles were administered as bolus intravenous infusions at a volume of 0.2 ml per 100 g body weight via a jugular vein catheter.

  Data and statistical analysis: These data were expressed as mean ± SEM for each group. Differences between groups were assessed for statistical significance by Student's T-test and by Dunnett's or Bonferroni's test for one-way or two-way ANOVA followed by optional multiple comparisons. A level of p <0.05 was considered significant. In addition, data on the effects of these multiple doses were evaluated using linear regression analysis to measure significant differences between the slopes of the lines between treatments.

Experimental design:
Experimental Series 1 : Measure the acute effects induced by a single dose of ipamorelin in a rat model of POI.
Rats were allowed to acclimate to the facility for 7 days; catheter patency was maintained. Day 0: Rats were weighed and fasted from 8:00 am to 9:00 am. Day 1: From 6:00 am to 8:00 am, “runnin” surgical dye under isoflurane anesthesia was injected into the proximal colon at the end of the surgery.
Single dose treatment with ipamorelin or vehicle.
Observations in the cage (time to first fecal excretion; fecal excretion and food intake at 3, 6, 9 and 12 hours after surgery).
・ Day 2: From 7:00 am to 8:00 am, body weight, fecal excretion, food intake at 24 hours after surgery ・ Day 3: From 7:00 am to 8:00 am, 48 hours after surgery Body weight, fecal excretion, dietary intake of foods were over-administered with isoflurane.

Inexperienced rats were fasted but did not undergo surgery. Body weight, large intestine passage, fecal excretion and food intake were measured at the same time as POI rats.
Group (single dose treatment):
POI + vehicle (intravenous administration) n = 12
POI + ipamorelin 0.1 mg / kg (intravenous administration) n = 9
POI + ipamorelin 1 mg / kg (intravenous administration) n = 10
POI + GHRP-6 20 μg / kg (intravenous administration) n = 8
Inexperienced (no surgery, no drugs) n = 14

Experimental Series 2 : Determine the efficacy of multiple doses of ipamorelin in a rat model of POI.
Rats were allowed to acclimate to the facility for 7 days; catheter patency was maintained. Day 0: Rats were weighed and fasted from 8:00 am to 9:00 am. Day 1: From 6:00 am to 8:00 am, “intestinal motility” surgical dye under isoflurane anesthesia was injected into the proximal large intestine at the end of the surgery.
Multiple doses of ipamorelin or vehicle (first dose at the end of surgery, second, third and fourth doses at 3 hour intervals).
Observations in the cage (time to first fecal excretion; fecal excretion and food intake at 3, 6, 9 and 12 hours after surgery).
• Day 2: From 7:00 am to 8:00 am, measure body weight 24 hours after surgery Multiple doses of ipamorelin or vehicle (first dose at weight measurement, second at 3 hour intervals) , Third and fourth doses).
Observation in cages (feces excretion and food intake at 24, 27, 30, 33 and 36 hours after surgery).
・ Day 3: From 7:00 am to 8:00 am, body weight, fecal excretion and food intake were measured 48 hours after surgery. Euthanasia by overdose of isoflurane.

Inexperienced rats were fasted but did not undergo surgery. Body weight, large intestine passage, fecal excretion and food intake were measured at the same time as POI rats. Note that naïve rats did not receive these treatments, unlike POI rats that were treated multiple times during the administration of ipamorelin or vehicle.
Group (multiple doses):
POI + vehicle (intravenous administration) n = 8
POI + ipamorelin 0.01 mg / kg (intravenous administration) n = 8
POI + ipamorelin 0.1 mg / kg (intravenous administration) n = 8
POI + ipamorelin 1 mg / kg (intravenous administration) n = 7
Inexperienced (no surgery, no drugs) n = 8

result
POI in rats : Effects of abdominal surgery on colon transit, fecal excretion, food intake and body weight.
A comparison between rats undergoing “intestinal motility” surgery and inexperienced rats shows that surgery delayed the passage of the large intestine (FIG. 12A), decreased fecal excretion (FIG. 12B), decreased food intake ( FIG. 12C) and the occurrence of POI characterized by a decrease in weight gain (FIG. 12D) were shown.

POI rat model. (A) Large intestine transit time (measured as time to excrement with the first mark) is significantly delayed after surgery compared to naive rats. (B) Cumulative excretion at 24 and 48 hours after surgery is reduced compared to naive rats. (C) Cumulative food intake is significantly reduced compared to naive rats at 24 and 48 hours after surgery. (D) Weight gain decreases after surgery compared to naive rats. Data are mean ± SEM from 12 POI rats and 14 naive rats. The significance of the difference between surgically and naïve rats was examined using Student's t-test: ^* p <0.05, ^** p <0.01, ^*** p <0. 001.

Experimental Series 1 : Measure acute effects induced by single dose treatment of ipamorelin in a rat model of POI.
Experiments were performed in POI rats to determine the efficacy of a single dose treatment of 0.1 mg / kg or 1 mg / kg ipamorelin. Ipamorelin or vehicle was administered by intravenous infusion following the end of surgery. The time to first fecal discharge (Figure 13) and cumulative fecal discharge (Figure 14), food intake (Figure 15) and weight gain (Figure 16) were measured at 48 hours after surgery, The efficacy of ipamorelin was evaluated by comparing the effect of ipamorelin with that of the vehicle. In addition, the potency of 20 μg GHPR-6 (an agonist of GRLN, ie GHS-R1a) was used as a positive control (Davenport et al., 2005). The results presented in FIG. 13 show that a single post-operative dose of 1 mg / kg ipamorelin or 20 μg GHRP-6 significantly reduced colon transit time.

  However, neither ipamorelin nor GHRP-6 had a significant effect on fecal excretion (FIG. 14), food intake (FIG. 15) or weight gain (FIG. 16) after the first 48 hours after surgery. It was.

  In summary, these results from Experimental Series 1 show that treatment with a single dose of 1 mg / kg ipamorelin given at the end of surgery was the first in the 48 hour recovery process in POI rats. It shows that the time to bowel movement was reduced but did not induce effects on fecal excretion and food intake. However, these results are consistent with what is expected based on the reported half-life of ipamorelin in rats of 30-60 minutes (Johansen et al., 1998).

Experimental Series 2 : Determine the efficacy of multiple doses of ipamorelin in a rat model of POI.
The dose-response effect of multiple doses of ipamorelin (0.01, 0.1 or 1 mg / kg, iv) was investigated in POI rats. The results showed that multiple doses of 0.1 mg / kg or 1 mg / kg ipamorelin caused significant acceleration compared to the effect of the vehicle through the large intestine (FIG. 17).

  The effect of multiple doses of ipamorelin on fecal excretion is presented in FIG. Multiple doses of ipamorelin induced an increase in cumulative fecal output compared to vehicle. This effect was significantly reached at doses of 0.1 mg / kg or 1 mg / kg (FIG. 18). Fecal output is higher (lower 1 / gradient value) in rats receiving ipamorelin at doses of 0.1 or 1 mg / kg (intravenous administration) compared to rats treated with vehicle Increased in At a dose of 0.01 mg / kg, ipamorelin had no significant effect. In addition, ipamorelin induced increased food intake (FIG. 19). Increased food intake induced by higher doses of ipamorelin was associated with weight gain. As shown in FIG. 20, rats receiving the highest dose of 1 mg / kg ipamorelin administered according to the multiple dose paradigm were compared to rats treated with vehicle in the first 48 hours after surgery. Significantly gained more body weight.

CONCLUSION Overall, these results show that multiple intravenous administration of ipamorelin in the first 48 hours after surgery improves colonic transit and food intake and increases body weight gain in rats. This suggests that intravenous infusion after ipamorelin surgery may improve symptoms in patients with POI.

  Now that the present disclosure has been fully provided, those skilled in the art will be able to do undue experimentation on the same in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of this disclosure. I will admit that I can do it without it. Moreover, while this disclosure has been described with respect to particular embodiments, it will be understood that further modifications are possible. This application is intended to cover any variations, uses, or adaptations of this disclosure, which are generally intended to cover such deviations from the disclosure in accordance with the principles disclosed. Are within the known or customary practice in the field to which this disclosure belongs and may be applied to the requisite matters indicated above.

References
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Masuda Y., Tanaka T., Inomata N, Ohnuma N., Tanaka S., Itoh Z., et al. Ghrelin stimulates gastric acid secretion and motility in rats. Biochem Biophys Res Commun 2000 Oct 5; 276 (3): 905 -8.
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Claims (18)

A method of treating a disease in a patient in need of treatment of a gastrointestinal disease, wherein the therapeutically effective amount of the ghrelin mimetic compound represented by Structural Formula I:

Or a method comprising administering a pharmaceutically acceptable salt, solvate or hydrate thereof.   The method of claim 1, wherein the patient is using opioids for management of pain after surgery.   The method of claim 1, wherein the patient is using an opioid for the management of chronic pain.   4. The method of claim 3, wherein the opioid is selected from the group consisting of morphine, codeine, oxycodone, hydromorphone, hydrocodone, methadone, and fentanyl. A method wherein a therapeutically effective amount of a laxative, peripherally acting opioid antagonist, proton pump inhibitor, H ₂ antagonist, antacid, serotonin receptor agonist (pure or mixture), motilin receptor agonist, dopamine antagonist, 2. The method of claim 1, further comprising administering a cholinesterase inhibitor, or a combination thereof.   The method of claim 1, wherein the patient is a human.   2. The method of claim 1, wherein the ghrelin mimetic is administered in a manner that achieves an effective plasma concentration.   2. The method of claim 1, wherein the gastrointestinal disease is gastrointestinal dysfunction induced by opioids.   2. The method of claim 1, wherein the gastrointestinal disorder is morphine-induced constipation.   The gastrointestinal disorder is opioid-induced constipation, diabetes-related or idiopathic gastric paresis, gastroesophageal reflux disease, irritable bowel syndrome, constipation, or postoperative ileus. the method of. A method for stimulating gastrointestinal motility in a patient in need of stimulating gastrointestinal motility, wherein the patient has a therapeutically effective amount of a ghrelin mimetic compound represented by Structural Formula I:

Or a method comprising administering a pharmaceutically acceptable salt, solvate or hydrate thereof.   The method of claim 11, wherein the patient is a human.   12. The method of claim 11, wherein the patient has opioid-induced constipation, diabetes-related gastric failure, gastroesophageal reflux disease, irritable bowel syndrome, constipation, or postoperative ileus.   12. The method of claim 11, wherein the ghrelin mimetic is administered in a manner that achieves an effective plasma concentration. Need to treat a disease or condition selected from the group consisting of opioid-induced constipation, diabetes-related or idiopathic gastric failure, gastroesophageal reflux disease, irritable bowel syndrome, constipation, and postoperative ileus A method of treating the disease or condition in a patient having a therapeutically effective amount of a ghrelin mimetic compound represented by Structural Formula I below:

Or a method comprising administering a pharmaceutically acceptable salt, solvate or hydrate thereof.   The method of claim 15, wherein the patient is a human.   16. The method of claim 15, wherein the ghrelin mimetic is administered in a manner that achieves an effective plasma concentration. A therapeutically effective amount of a laxative, a peripherally acting opioid antagonist, further comprising a proton pump inhibitor, H ₂ antagonists, antacids, serotonin receptor agonists, motilin receptor agonists, administering a dopamine antagonist, or a cholinesterase inhibitor, claim 15. The method according to 15.

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