JP2007538017A - PPY-containing composition for the treatment of gastrointestinal diseases - Google Patents

Abstract

The present invention relates to PYY or a functional equivalent thereof used in a pharmaceutical composition. The pharmaceutical compositions are particularly useful for the treatment of functional gastrointestinal diseases such as irritable enteritis and functional dyspepsia. The present invention further relates to a method of treatment using the composition. Furthermore, the combination of PYY or a functional equivalent thereof and a second active ingredient such as an antiemetic is also included.

Description

  The present invention relates to PPY-containing compositions for the treatment of gastrointestinal diseases. The composition of the present invention is for the manufacture of a pharmaceutical composition for the treatment of IBS, FD and / or abdominal pain. The invention further relates to a method of treatment characterized in that the pharmaceutical composition is administered alone or in combination with a second active ingredient.

  Gastrointestinal disease is very common in the population. Some of these are very characteristic and therefore suitable treatment regimes have been developed. Development of methods for treating functional gastrointestinal diseases is even more difficult. The term refers to a disease or disorder in which the major abnormality is a change in physiological function (how the body functions) rather than an identifiable structural or biochemical cause. Thus, the cause of these types of gastrointestinal diseases is unknown, meaning that there is no biological explanation. In general, this type of disease cannot be diagnosed by traditional methods, i.e., as commonly used diagnostic methods, X-rays, or inflammation, infection, or structural abnormalities that can be seen in clinical laboratory tests. Cannot diagnose.

  Gastrointestinal diseases of unknown cause include irritable bowel syndrome (IBS), also called irritable colon, functional dyspepsia (FD) or non-ulcer dyspepsia. Therefore, the diagnosis of this category of gastrointestinal diseases and the development of suitable treatment methods have not been fully successful at present.

The main symptoms of irritable bowel syndrome IBS are abdominal pain, alternating stool habits, defecation difficulties and discomfort associated with abdominal distension. Patients have increased mucus, nausea, and constipation and bloating. The diagnostic criteria for irritable bowel syndrome are according to ROME II (see below), supervised by Talley, NJ and Spinller R (2002) and Talley, NJ (2003). It occupies a week and is accompanied by two or more of the following characteristics: lightness due to defecation, softening or increasing the frequency of stool, hardening or decreasing the frequency of stool. Symptoms are chronic and impair the patient's quality of life.

IBS ROME II Symptom Diagnosis Criteria Of the past 12 months, it is not necessarily continuous, but there is abdominal discomfort or abdominal pain for at least 12 weeks, and hypersensitivity to two of the following three features Sexual bowel syndrome:
(1) ameliorates after defecation; and / or (2) there is a change in the number of defecations at onset; and / or (3) there is a change in fecal characteristics at the onset, but not essential, but others that help diagnose IBS As symptoms of
Abnormal number of bowel movements (defecation frequency is 3 times a day or less than 3 times a week);
Abnormal stool properties (feces or hard stools, soft stools (mud) or watery stools);
Abnormal defecation (straining, tightness of stool or feeling of residual stool);
Mucus adherence during defecation;
Fullness or abnormal abdominal distension It is estimated that 10-15% of the population experience IBS symptoms.

Patients are divided into three groups based on primary stool habits (diarrhea and / or constipation).
IBS with abdominal pain, constipation and diarrhea
IBS with abdominal discomfort, bloating and constipation
IBS with alternating diarrhea and constipation

Functional dyspepsia Functional dyspepsia (FD) symptoms partially overlap with IBS symptoms. Indigestion means persistent or frequent upper abdominal pain or subjective upper abdominal discomfort, characterized by an early feeling of fullness, a feeling of fullness after meals, or a feeling of fullness. Indigestion is not always a diet-related symptom. Patients are divided into movement disorder-like FD and ulcer-like FD (discomfort and pain, respectively) based on general symptoms centered on the upper abdomen. Many patients experience symptoms including upper abdominal discomfort, satiety and pain after digestion. Additional symptoms include inability to eat a standard amount of food, bloating, nausea, nausea and vomiting (Feinle-Bisset, C et al., 2003). The underlying mechanism of functional dyspepsia is unknown. The role of delayed gastric emptying has been discussed and is not currently considered a preferred model.

Central nervous system, visceral hypersensitivity and stress brain-gut interactions play a major role in the regulation of gastrointestinal function in health and disease. The central nervous system (CNS) regulates gastrointestinal functions such as movement, secretion and blood flow. This happens without the individual being aware. In the case of irregular gastrointestinal tracts, individuals experience short periods of pain. For patients with gastrointestinal disease, the pain is very strong and long. This appears to be associated with visceral hypersensitivity in the pathophysiology of functional gastrointestinal diseases including IBS and FD (Feinle-Bisset, C et al., 2003). Furthermore, the recall of stress, anxiety, or repellent memory enhances the recognition of painful events. Thus, stress has the effect of inducing hyperalgesia (Drossman DA, et al., 2002).

Psychological factors Psychological and sociological factors appear to be related to the onset and severity of symptoms (Drossman DA, et al., 2002), and the most effective drug for the treatment of irritable bowel syndrome is currently antidepressant Seems to be an agent (Talley, NJ and Spiller R).

Treatment At present, the treatment of gastrointestinal disorders of unknown cause, such as irritable bowel syndrome and functional dyspepsia, is based on empirical facts and aims to regulate symptoms. The complexity of the symptoms suggests that a single treatment method is not effective for all patients.

  Symptoms related to diarrhea and constipation are treated with constipation drugs and antidiarrheal drugs, but the success is minimal. In addition, drugs such as loperamide can minimize diarrhea, but do not relieve abdominal pain (reviewed by Talley, NJ 2003), which means that irregular stool habits and experienced abdominal pain Suggests that they are not directly related.

  As noted above, antidepressants have some effect on IBS, and most patients with diarrhea with IBS are treated with tricyclic antidepressants such as desipramine, nortyptiline, fluphenzine, etc. In clinical trials, the quality has changed. Selective serotonin reuptake inhibitors (SSRIs) are expected to be effective in constipated IBS patients (reviewed by Talley, N.J., 2003).

Gastrointestinal hormones Several gastrointestinal hormones have been suggested to be associated with symptoms of gastrointestinal disease. VIP, CCK and motilin are involved in upper gastrointestinal motility, and polypeptide YY (PYY) and neuropeptide Y (NPY) affect intestinal absorption. The latter (both PYY and NPY) has been proposed as a treatment for diarrhea by extending the residence time (US 6,588,708).

PYY
Gastrointestinal hormone peptide YY (PYY) and the neuropeptide, neuropeptide Y (NPY) are structurally related to pancreatic polypeptide (PP) (FIG. 1). PYY and NPY exert their actions via NPY receptors (Y1R, Y2R, Y4R and Y5R). PP, NPY and PYY peptides consist of 36 amino acids with an amidated C-terminus. Two forms of PYY, PYY1-36 and PYY3-36, have been found in the blood circulation, although the latter is a truncated form of the former. PYY3-36 is made by cleaving PYY1-36 with the enzyme dipeptidyl peptidase IV (DPP-IV). PYY1-36 binds to and activates at least three NPY receptor subtypes (Y1, Y2, and Y5), while PYY3-36 is more selective for the Y2 receptor (Y2R). Only the C-terminal part of the PYY3-36 peptide is required for binding to the Y2 receptor. Throughout this document, the term PYY encompasses both PYY1-36 and PYY3-36 (Berglund, MM et al., 2003).

PYY was first isolated from porcine stomach (Tatemoto, K and Mutt, C., 1980) and named peptide YY based on the tyrosine residues present at the N- and C-termini of the molecule.
PYY is expressed in endocrine cells lined up in the gastrointestinal tract, particularly in the distal region. PYY is secreted in response to food digestion. Within 15 minutes, the plasma level of PYY rises and the PYY level reaches a plateau after about 90 minutes. The maximum level of PYY reached is proportional to the calories burned, suggesting that PYY will function as a sensor for food digestion. Furthermore, PYY is also expressed by nerve cells such as peripheral neurons, particularly intestinal neurons. Furthermore, PYY is found in a limited set of central nerves. Expression patterns of PYY both in endocrine cells and in neurons suggest that PYY may be involved in the regulation of diverse functions in individuals (Ekblad, E. and Sundler, F., 2002)

  One proposed role for PYY would be to regulate fluid and electrolyte secretion and absorption in the gastrointestinal tract and intestine. Therefore, PYY has been proposed as a therapeutic agent for diarrhea by extending the residence time (US 6,588,708). Furthermore, it has been suggested that PYY3-36 may be involved in a system that regulates feeding behavior. It has been discovered that peripheral administration of PYY3-36 inhibits food intake in rodents. Furthermore, direct administration of PYY3-36 into the intra-arcuate inhibits food intake. The relationship between the effects of PYY on feeding behavior and the NPY2 receptor is suggested by NPY receptor Y2-deficient mice being resistant to the appetite suppressive effect of peripheral administration of PYY3-36 (Batteham, RL and Bloom, SR, 2003).

  The hypothalamic arcuate nucleus, an important brain region that regulates appetite, can utilize nutrients and hormones in the peripheral circulation. NPY neurons in the hypothalamic nucleus express Y2R. The cone nucleus contains two distinct subgroups of neurons that control food intake. One group of neurons produces NPY that acts on the brain to stimulate feeding (Stanley, BG et al., 1986), and adjacent subgroups of neurons act in the same brain area as NPY, but Produces melanocortin peptides that inhibit food (Fan, W. et al., 1997). In general, when one of these subsets is activated, the other is inhibited.

References
Batterham, RL and Bloom, S, R (2003): Gastrointestinal hormone peptide YY regulates appetite: Ann. NY, Acad. Sci. June; 994: 162-8
Berglund, MM; Hipskind, PA and Gehlert, DR (2003): Recent progress in our understanding of the physiological role of PP-fold peptide receptor subtypes: Exp Biol Med (Maywood), Mar; 228 (3): 217-44
Drossman, DA; Camilleri, M .; Mayer, EA; Whitehead WE (2002): AGA technical examination of irritable bowel syndrome: Gastroenterology, Dec; 123 (6): 2108-31
Ekblad, E. and Sundler, F. (2002): Distribution of pancreatic polypeptide and peptide YY: Peptides. Feb: 23 (2): 251-61
Fan. W .; Boston, BA; Kesterson, RA: Hruby, VJ; Cone, RD (1997): Role of melanocortinic neurons in feeding and Agouti obesity syndrome: Nature. Jan 9; 385 (6612): 165- 168
Feinle-Bisset, C .; Vozzo, R .; Horowitz, M., and Talley, NJ (2003): American Journal of Gastroenterology, Jan: 99 (1): 170-181
Stanley, BG; Kyrkouli, SE; Lampert, S. and Leibowitz, SF (1986): Peptides 7, 1189-1192)
Talley, NJ and Spiller R (2002): Irritable bowel syndrome: Slightly understood organ bowel disease: The Lancet, Vol. 360, August 17, 555-564
Talley, NJ (2003): Evaluation of drug treatment for irritable bowel syndrome: J. Clin. Pharmacol, 56, 362-369
Tatemoto, K. and Mutt, V. (1680): Isolation of two new hormone candidates using chemical discovery methods for naturally occurring polypeptides: Nature, June 5; 285 (5764): 417-8

The present invention relates to a PYY-containing composition for the manufacture of a medicament for the treatment of functional gastrointestinal diseases. The drug is a) constipation with primary stool habits, and / or b) diarrhea and constipation with primary stool habits alternating, and / or c) treatment with parenteral administration,
Can be used to treat irritable bowel syndrome.

  The medicament is for the treatment of abdominal and visceral pain associated with functional gastrointestinal diseases such as irritable bowel syndrome and functional dyspepsia.

Furthermore, the medicament is for the treatment of functional dyspepsia for the treatment of the following symptoms:
a) satiety, and / or b) unable to eat a standard amount of meal, and / or c) pain after food intake, and / or d) nausea and / or e) vomiting, and / or f A) abdominal distension, and / or g) stagnation, and / or h) reflux, and / or i) upper abdominal pain, and / or j) bloating, and / or k) excess intestinal gas,
And any combination of the above symptoms.

  The present invention further relates to a method of treatment characterized by administering the composition.

Definition
AA : See “Amino acid”
Amino acid : An entity comprising an amino terminal part (NH ₂ ) and a carboxy terminal part (COOH) separated by a carbon atom or a central part containing a carbon atom chain, and having at least one side chain or functional group. NH ₂ means an amino group present at the amino terminus of an amino acid or peptide, and COOH means a carboxyl group present at the carboxy terminus of an amino acid or peptide. The general term amino acid encompasses both natural and unnatural amino acids. Natural amino acids of the standard nomenclature listed in J. Biol. Chem., 243: 3552-59 (1969) and adopted in 37 CFR, section 1.822 (b) (2) are listed in Table 1 below. It belongs to the amino acid group. Unnatural amino acids refer to those not listed in Table 1. Examples of unnatural amino acids are those listed in 37 CFR, section 1.822 (b) (4), all of which are incorporated herein by reference. The amino acid residues described herein can be both “D” isomers and “L” isomers.

Amino acid residues : The term “amino acid residue” includes standard amino acids, non-standard amino acids, pseudo-amino acids, which are at least one other species, such as 2, eg 3, Or it reacted with 3 or more other species. In particular, the amino acid residue has an acyl bond instead of a free carboxyl group and / or an amine bond and / or an amide bond instead of a free amine group. Furthermore, the reacted amino acid residue has an ester or thioester bond instead of an amide bond.

Antibody : An antibody refers to an immunoglobulin molecule and the active part of an immunoglobulin molecule. An antibody is, for example, an intact immunoglobulin molecule or a fragment thereof that retains immune activity.

Antigen : A molecule recognized by an antibody. Usually a peptide, porpeptide or multimeric polypeptide. The antigen preferably has the ability to elicit an immune response.

Chimera : A molecule that consists of at least two parts that are not found together in nature. A chimeric peptide or protein is a peptide or protein constructed by fusing two peptides or proteins. A chimeric DNA molecule is a DNA molecule that encodes a chimeric protein.

Concentration equivalent : a dose equivalent defined as the dose of a compound that exhibits the same response in vitro and / or in vivo as a known compound (evaluated from a dose-response curve, etc.) It is.

Frequency : The number of times an event occurs within a period (for example, the number of occurrences per day or week).

Individual : A living animal or human. In preferred embodiments, the subject is a mammal, including humans and non-human mammals, such as dogs, cats, pigs, cows, sheep, goats, horses, rats and mice. In the most preferred embodiment, the subject is a human.

Isolation : Used to describe the various secretagogues, polypeptides, and nucleotides disclosed herein that have been identified and separated and / or recovered from components of their natural environment. Impure components of its natural environment are materials that generally inhibit the diagnostic or therapeutic use of polypeptides, including enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In a preferred embodiment, the polypeptide will be purified.

Ligand : A molecule, such as a peptide, that can specifically bind to one or more cognate receptors. An antigen is, for example, a ligand for its cognate antibody.

Medical disorder:
By the term “medical disease” is meant any disease or symptom that has a detrimental effect on an individual's physical and / or mental health.

MTD : Maximum tolerance. The maximum tolerated dose of a substance during a given test period is: a) obvious toxicity, eg, obvious cell death or organ dysfunction; b) substantial reduction in lifespan other than that caused by cancer; c) growth phase Do not induce 10% or more inhibition of weight gain in animals.

Parenteral : For the purposes of the present invention, “parenteral” means outside the gastrointestinal tract. Thus, the term “parenteral administration” of a compound includes, for example, subcutaneous, intramuscular, intravenous, nasal, buccal, intradermal and transdermal administration, and also includes inhalation of the compound.

Peptide : A plurality of covalently linked amino acid residues that determine the sequence, linked by amino bonds. The term is used in the same way as oligopeptide and polypeptide. Amino acids include both natural and unnatural amino acids, and combinations thereof. Natural and / or non-natural amino acids are linked by peptide bonds or non-peptide bonds. The term peptide also encompasses post-translational modifications known in the art introduced by chemical or enzyme catalysis.

PYY : Peptide YY. Here, PYY means both PYY1-36 and PYY3-36, and includes PYY having a full length and a shape in which the tip is cut off, respectively.

Receptor : A receptor is a molecule that binds specifically (not randomly) to another molecule, such as a protein or glycoprotein.

Recombinant DNA (rDNA) molecule : A DNA molecule produced by linking two DNA fragments in an operable manner. Thus, a recombinant DNA molecule is a hybrid DNA molecule having at least two nucleotide sequences that are not normally found together in nature.

T _1/2 :
Half-life is the time for the compound concentration to decrease by 50%.

[Detailed description of drawings]
Figure 1:
The PP-fold peptide family structures NPY, PYY and PP share a common hairpin-like three-dimensional structure called PP-fold (Fuhlendorf, J. et al. (1990) J. Biol. Chem. ). All four peptides are 36 amino acids long with an amidated carboxy terminus. The general structure of PP-fold peptides was defined using X-ray crystallography of avian PP and confirmed in several experiments using nuclear magnetic resonance (Keire DA et al. (2000) Biochemistry). Amino acid residues 1-8 form a type II proline helix followed by a loop. Residues 15-32 form an α-helix and the four carboxy terminal residues are in a flexible loop conformation. NPY is one of the most evolutionarily conserved known peptides. Despite the low degree of amino acid sequence conservation between PP and other different species, and between PP, PYY and NPY, the overall three-dimensional structure appears to be conserved in all PP fold peptides It is.

Figure 2:
A PYY plasma concentration PPY1-36 in response to PPY1-36 subcutaneous administration was administered to the subject as described in Example 3. Plasma levels of PYY were measured 4 hours after injection. Plasma levels of PYY increase within 15 minutes after administration. Plasma levels of 80-100 pmol / l are obtained at a dose of 200 pmol / kg (FFM).

Figure 3:
A PYY plasma concentration PPY3-36 in response to PPY3-36 subcutaneous administration was administered to the subject as described in Example 3. Plasma levels of PYY were measured 4 hours after injection. Plasma levels of PYY increase within 15 minutes after administration. Plasma levels of 100-120 pmol / l are obtained at a dose of 100 pmol / kg (FFM)

Detailed Description of the Invention
Gastrointestinal disease:
The gastrointestinal diseases described in the background section of this application are characterized by unknown causes and are later diagnosed based on patient symptom reports. The main symptoms are abdominal discomfort and abdominal pain with irregular bowel movements.
Symptoms of the patient are constipation, diarrhea, abdominal pain, fullness, inability to eat a standard amount of food, abdominal distension, vomiting, nausea, and upper abdominal pain.

Individuals who need:
According to the present invention, any individual who can derive the effect of the composition of the present invention can be treated with the composition. Therefore, an individual who brings out this therapeutic effect is regarded as an individual who needs treatment.
Preferably, the individual is suffering from a gastrointestinal disorder. The individual in need suffers from one of the above diseases. The individual in need is one who suffers from any of the above symptoms of the disease, such as abdominal pain, visceral pain, abdominal discomfort, diarrhea, constipation, nausea, vomiting, abdominal distension, snoring or abdominal distension.
An individual in need is an individual who is at risk of suffering from any of the above diseases.
As described above, the symptoms associated with such gastrointestinal diseases are difficult to treat. Surprisingly, the present invention has succeeded in treating gastrointestinal diseases by using PYY and its functional equivalents.

PYY:
Throughout this document, peptide YY (PYY) is used as a general term encompassing both PYY1-36 and PYY3-36. PYY1-36 is a 36AA polypeptide with C- and N-terminal tyrosine amino acid residues. The polypeptide is prepared by cleaving the prepolypeptide and further cleaved by depeptidyl peptidase IV as described above to produce PYY3-36. Surprisingly, it has continued to be found that PYY has the ability to suppress several symptoms of gastrointestinal diseases such as IBS and functional dyspepsia.

Functional equivalent:
Human PYY1-36 is identified by SEQ ID NO: 1 (SEQ ID NO: 1). PYY3-36 is a truncated form of PYY in which the two N-terminal residues are removed. Functional equivalents of PYY include PYY molecules originating from different species, eg, mouse, rat, monkey, pig, cow, or other mammalian species. Functional equivalents include homologues of PYY.
The present invention relates to a composition containing PYY or a functional equivalent thereof for the preparation of a pharmaceutical composition for the treatment of gastrointestinal diseases of unknown cause.

In one aspect of the invention, the composition is for the preparation of a pharmaceutical composition for the treatment of gastrointestinal diseases of unknown cause characterized by the following symptoms:
a) constipation, and / or b) diarrhea, and / or c) abdominal pain.

In one aspect, the composition containing PYY or a functional equivalent thereof is for the preparation of a pharmaceutical composition for the treatment of irritable bowel syndrome with the following symptoms:
a) primary stool habit is constipation, and / or b) diarrhea and constipation, where main stool habit is alternating, and / or c) treatment is parenteral

In a further embodiment, the composition is for the treatment of functional dyspepsia.
In a further embodiment, the composition is for the treatment of abdominal pain, visceral pain, eg left upper abdominal pain, upper right abdominal pain, left lower abdominal pain, right lower abdominal pain.
In one aspect, the composition is therapeutic.
In one embodiment, the composition is for the treatment of pain associated with IBS or functional dyspepsia.
In one embodiment, the composition is for the treatment of the following symptoms:
l) satiety, and / or m) unable to eat a standard amount of meal, and / or n) pain after food intake, and / or o) nausea, and / or p) vomiting, and / or q A) abdominal distension, and / or r) snoring, and / or s) reflux, and / or t) upper abdominal pain, and / or u) bloating, and / or v) excess intestinal gas,
And any combination of the above symptoms.

  In a preferred embodiment, the composition comprises human PYY. In a further preferred embodiment, the composition comprises human PYY1-36 as defined in SEQ ID NO: 1. In a further preferred embodiment, the composition comprises human PYY3-36 defined by amino acids 3-3 of SEQ ID NO: 1.

Homologue:
Homologues are herein interpreted as molecules that share some identity to the PYY molecule represented by both PYY1-36 and PYY3-36. Homology aligns sequences, if necessary, fills gaps to achieve maximum percent identity with the entire sequence, but does not consider any conservative substitutions as part of sequence identity, comparisons in candidate sequences Expressed as a percentage of amino acid residues identical to the corresponding sequence residues. N- or C-terminal extensions or insertions are not considered to reduce identity or homology. Methods and computer programs for alignment are well known in the art. Sequence identity can be measured using sequence analysis software (eg, Sequence Analysis Software Package, Genetics Computer Group, University of Wisconsin Biotechnology Centre, 1710 University Ave., Madison, Wis. 53705). The software assigns varying degrees of homology to various substitutions, deletions, and other modifications to match similar sequences.

  One or more homologues of the identified sequences differ in one or more amino acids compared to the defined sequence, but can perform the same function, ie, the homologues are of a predetermined sequence It can be conceived as a “functional equivalent”.

As noted above, the functional equivalent of any given sequence can be defined as follows:
i) a homolog having an amino acid sequence that can be recognized by an antibody, which can also recognize PYY (PYY1-36 and / or PYY3-36), and / or ii) select for the NPY receptor Homologues having an amino acid sequence that can bind manually (eg, as measured in Example 1), and / or iii) substantially equivalent to PYY (PYY1-36 or PYY3-36) in the NPY receptor, or Homologues with a higher degree of binding affinity, preferably showing a binding affinity higher than 100 nM, and / or iv) a homologue showing at least 60% identity to human PYY identified by SEQ ID NO: 1 And / or v) a homologue consisting of a fragment of human PYY identified by SEQ ID NO: 1, wherein the fragment is a minor sequence of SEQ ID NO: 1 A homolog having at least 6 consecutive stretches of amino acids.

  Human PYY1-36 has the sequence of SEQ ID NO: 1. Human PYY3-36 is 34 amino acids long and has the sequence of SEQ ID NO: 1 except that the two N-terminal amino acids of SEQ ID NO: 1 are deleted.

  Examples of homologues include substitution of one or more conservative amino acids, including one or more conservative amino acid substitutions within the same group as a given amino acid, or each conservative substitution differs from a defined amino acid Includes multiple conservative amino acid substitutions, with substitutions within the group.

  Thus, a homologue includes conservative substitutions independent of each other, wherein at least one glycine (Gly) of the homologue is substituted with an amino acid independently selected from the amino acid group consisting of Ala, Val, Leu, and Ile. At least one alanine (Ala) of the homologue is substituted with an amino acid independently selected from the amino acid group consisting of Gly, Val, Leu, Ile, at least one valine (Val of the homologue) ) Is substituted with an amino acid independently selected from the amino acid group consisting of Gly, Ala, Leu and Ile, and at least one of the leucine (Leu) of the homologue consists of Gly, Ala, Val and Ile One substituted with an amino acid selected independently from the amino acid group, at least one of the homologous isoleucine (Ile) In which at least one of the aspartic acid (Asp) of the homologue is Glu, Asn, Gln is substituted with an amino acid independently selected from the amino acid group consisting of Gly, Ala, Val, Leu Wherein at least one of the phenylalanine (Phe) of the homologue is an amino acid independently selected from the amino acid group consisting of Tyr, Trp, His, Pro, preferably Substituted with an amino acid independently selected from the amino acid group consisting of Tyr, Trp, and at least one of the tyrosine (Tyr) of the homologue is independent of the amino acid group consisting of Phe, Trp, His, Pro Selected amino acids or amino acids independently selected from the amino acid group consisting of Phe and Trp An acid-substituted one, at least one of the arginine (Arg) of the fragment substituted with an amino acid independently selected from the amino acid group consisting of Lys, His, and the homologous lysine (Lys) One in which at least one is substituted with an amino acid independently selected from the amino acid group consisting of Arg and His, and at least one of the asparagine (Asn) of the homologue is independent from the amino acid group consisting of Asp, Glu and Gln In which at least one of the homologous glutamine (Gln) is substituted with an amino acid independently selected from the amino acid group consisting of Asp, Glu and Asn, the homologue At least one proline (Pro) is independently selected from the amino acid group consisting of Phe, Tyr, Trp, and His. Independently of the amino acid group consisting of Asp, Glu, Lys, Arg, His, Asn, Gln, Ser, Thr, Tyr, wherein at least one of the cysteines (Cys) of the homologue is substituted And those substituted with a selected amino acid.

  Conservative substitutions can be introduced at any position of the preferred defined sequence. However, it is also desirable to introduce non-conservative substitutions, particularly but not limited to, non-conservative substitutions at any one or more positions.

  Non-conservative substitutions to form functionally equivalent homologues of the sequence include, for example, i) polar side chains such as Gly, Ser, Thr, Cys, Tyr, Asn or Gln that are substantially different in polarity. Or a charged amino acid such as Asp, Glu, Arg, or Lys with a residue having a non-polar side chain (Ala, Leu, Pro, Trp, Val, Ile, Leu, Phe or Met), or non- Substituting polar residues with charged or polar residues; and / or ii) substantially different in their effect on polypeptide backbone coordination such as substituting Pro or Gly with other residues: and / or iii ) Substituting positively charged residues such as Lys, His and Arg with negatively charged residues such as Glu and Asp (or vice versa) with substantially different electrical charges And / or iv) substituting residues with small side chains such as Ala, Gly or Ser with (or their) bulky residues such as His, Trp, Phe, Tyr, etc. that are substantially different in steric volume; Opposite).

  In certain embodiments, amino acid substitutions are performed based on their hydrophobicity and hydrophilicity, including charge, size, etc., and the relative similarity of amino acid side chain substitutions. Examples of amino acid substitutions that take into account one of the above features are well known to those skilled in the art, such as arginine and lysine; glutamate and aspartate; serine and threonine; glutamine and asparagine; Leucine and isoleucine.

  In a preferred embodiment, the homologue has an amino acid sequence that is at least 60% identical to SEQ ID NO: 1.

  More preferably, the identity is at least 65%, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95%, identical to SEQ ID NO: 1. For example, at least 98% match.

  In a preferred embodiment, the functional equivalent has an amino acid corresponding to the 6 N-terminal amino acids (Tyr Pro Ile Lys Pro Glu) of PYY1-36 as defined in SEQ ID NO: 1. Functional equivalents are the eight N-terminal amino acids of PYY1-36 (Tyr Pro Ile Lys Pro Glu Ala Pro) as defined in SEQ ID NO: 10 or 10 of PYY1-36 as defined in SEQ ID NO: N-terminal amino acid (Tyr Pro Ile Lys Pro Glu Ala Pro Gly Glu), or 12 N-terminal amino acids (Tyr Pro Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala) or sequence defined in SEQ ID NO: 14 N-terminal amino acids of PYY1-36 defined in the number (Tyr Pro Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro) or 16 N-terminal amino acids of PYY 1-36 defined in the sequence number ( Tyr Pro Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu) or 18 N-terminal amino acids of PYY1-36 defined in SEQ ID NO: Tyr Pro Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn), or 20 N-terminal amino acids of PYY1-36 as defined (Tyr Pro Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr) or 22 of PYY 1-36 as defined in SEQ ID NO: N-terminal amino acids (Tyr Pro Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala) or 24 N-terminal amino acids of PYY1-36 defined in SEQ ID NO: Tyr Pro Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Tyr Ala Ser Leu) or 26 N-terminal amino acids of PYY1-36 defined in SEQ ID NO: Tyr Pro Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His) or 28 N-terminal amino acids of PYY1-36 defined in SEQ ID NO: Tyr Pro Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu), or 30 N-terminal amino acids of PYY1-36 (Tyr Pro Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu Asn Leu) defined in the column number, or 32 N-terminal amino acids of PYY1-36 defined in SEQ ID NO: Tyr Pro Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu Asn Leu Val Thr Or 34 N-terminal amino acids of PYY1-36 defined in SEQ ID NO: Tyr Pro Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln).

  In a further preferred embodiment, the functional equivalent is the 6 N-terminal amino acids of PYY3-36 defined by SEQ ID NO: 1 (Ile Lys Pro Glu Ala Pro), or 8 of PYY3-36 defined by SEQ ID NO: N-terminal amino acid (Ile Lys Pro Glu Ala Pro Gly Glu) or 10 N-terminal amino acids of PYY3-36 (Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala) defined by SEQ ID NO. 12 N-terminal amino acids of PYY3-36 (Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro) or 14 N-terminal amino acids of PYY3-36 defined by SEQ ID NO: Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu), or 16 N-terminal amino acids of PYY3-36 defined by SEQ ID NO (Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn), or SEQ ID NO: P defined 18 N-terminal amino acids of Y3-36 (Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr) or PYY3-36 20 N-terminal amino acids (Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala) or 22 N-terminal amino acids of PYY3-36 defined by SEQ ID NO: Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ayr Ser Leu) or 24 N-terminal amino acids of PYY3-36 defined by SEQ ID NO: Tyr Pro Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His) or 26 N-terminal amino acids of PYY3-36 defined by SEQ ID NO: Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu), or PYY3-3 defined by SEQ ID NO: 28 N-terminal amino acids (Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu Asn Leu), or 30 of PYY3-36 defined by SEQ ID NO: N-terminal amino acids (Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu Asn Leu Val Thr) or 32 of PYY3-36 defined by SEQ ID NO: It includes amino acids corresponding to N-terminal amino acids (Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln).

  In a preferred embodiment, the functional equivalent is the 6 C-terminal amino acids of PYY1-36 defined in SEQ ID NO: 1 (Val Thr Arg Gln Arg Tyr), or 8 of PYY1-36 defined in SEQ ID NO: 1. C-terminal amino acid (Asn Leu Val Thr Arg Gln Arg Tyr), 10 C-terminal amino acids of PYY1-36 defined in SEQ ID NO: 1 (Tyr Leu Asn Leu Val Thr Arg Gln Arg Tyr), or SEQ ID NO: 1 Twelve C-terminal amino acids of PYY1-36 (Arg His Tyr Leu Asn Leu Val Thr Arg Gln Arg Tyr) or 14 C-terminal amino acids of PYY1-36 defined in SEQ ID NO: 1 (Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln Arg Tyr) or 16 C-terminal amino acids of PYY1-36 defined in SEQ ID NO: 1 (Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln Arg Tyr Tyr Ala), Or as defined in SEQ ID NO: 1 18 C-terminal amino acids of PYY1-36 (Arg Tyr Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln Arg Tyr Tyr Ala) or 20 C-terminal amino acids of PYY1-36 defined in SEQ ID NO: 1 (Leu Asn Arg Tyr Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln Arg Tyr Tyr Ala) or 22 C-terminal amino acids of PYY1-36 defined in SEQ ID NO: 1 (Glu Glu Leu Asn Arg Tyr Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln Arg Tyr Tyr Ala) or 24 C-terminal amino acids of PYY1-36 defined in SEQ ID NO: 1 (Ser Pro Glu Glu Leu Asn Arg Tyr Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln Arg Tyr Tyr Ala) or 26 C-terminal amino acids of Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln Arg Tyr Tyr Ala) or P defined in SEQ ID NO: 1 28 C-terminal amino acids of Y1-36 (Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln Arg Tyr Tyr Ala) or PYY1 as defined in SEQ ID NO: 1 -36 30 C-terminal amino acids (Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln Arg Tyr Tyr Ala) or as defined in SEQ ID NO: 1 32 C-terminal amino acids of PYY1-36 (Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln Arg Tyr Tyr Ala) 34 C-terminal amino acids (Tyr Ala Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln Arg Tyr) Has the corresponding amino acid.

  In other preferred embodiments, the functional equivalent is an internal amino acid of PYY1-36, such as amino acids 16-21 as defined in SEQ ID NO: 1 (Glu Leu Asn Arg Tyr Tyr), or PYY1-36 as defined in SEQ ID NO: 1. Amino acid 15-22 (Glu Glu Leu Asn Arg Tyr Tyr Ala), or PYY1-36 amino acid 14-23 (Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser) defined in SEQ ID NO: 1, or SEQ ID NO: 1 PYY1-36 amino acids 13-24 (Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu) as defined, or PYY 1-36 amino acids 12-25 (Ala Ser Pro Glu Glu Leu Asn as defined in SEQ ID NO: 1) Arg Tyr Tyr Ala Ser Leu Arg), or amino acids 11-26 of PYY1-36 defined in SEQ ID NO: 1 (Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Tyr Ala Ser Leu Arg His), or SEQ ID NO: 1 PYY1-36 amino acids 10-27 (Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr), or PYY 1-36 amino acids 9-28 (Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu) or amino acids 8-29 of PYY1-36 defined in SEQ ID NO: 1 (Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu Asn) or amino acids 7-30 of PYY1-36 defined in SEQ ID NO: 1 (Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu Asn Leu), or amino acids 6-31 of PYY1-36 defined in SEQ ID NO: 1 (Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu Asn Leu Val), or Amino acid 5 of PYY1-36 defined in SEQ ID NO: 1 32 (Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu Asn Leu Val Thr) or amino acids 4-33 of PYY1-36 defined in SEQ ID NO: 1 (Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg) or amino acid 3-34 of PYY1-36 defined in SEQ ID NO: 1 (Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln) or amino acids 2-35 of PYY1-36 defined in SEQ ID NO: 1 (Pro Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln Arg), or amino acid 2- of PYY1-36 defined in SEQ ID NO: 1 36 (Pro Ile Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Le u Arg His Tyr Leu Asn Leu Val Thr Arg Gln Arg Tyr) or amino acids 4-36 of PYY1-36 defined in SEQ ID NO: 1 (Lys Pro Glu Ala Pro Gly Glu Asp Ala Ser Pro Glu Glu Leu Asn Arg Tyr Tyr Ala Ser Leu Arg His Tyr Leu Asn Leu Val Thr Arg Gln Arg Tyr).

  In certain embodiments, the functional equivalent has any of the above sequences with conservative amino acid substitutions, eg, 1 substitution, eg 2 substitutions, eg 2 substitutions, eg 2 or more substitutions, eg 4 or more substitutions.

  Also included are functional equivalents known in the literature, such as the PYY agonists disclosed in WO 03/057235 and references therein.

Covalently modified functional equivalents encompass all types of modifications. Nearly 200 structurally distinguishable covalent modifications of varying size and complexity ranging from amide to carboxylic acid conversion to adhesion to multiple complex oligosaccharides have now been identified. Such modifications include phosphorylation, acetylation, ubiquinoneation, lipidation (acetylation, phenylation, farnesylation, geranylation, palmitoylation, myristoylation), methylation, carboxylation, sulfuration, and O- Or N-glycosylation.

  Some modifications depend on vitamin C as a cofactor. This includes proline and lysine hydroxylation and carboxy-terminal amidation.

  In one embodiment, PYY or functional equivalent thereof includes C-terminal amidation. In a preferred embodiment, the C-terminal tyrosine residue of PYY or a functional equivalent thereof is amidated.

Functional equivalents according to the protecting group present invention has a protecting group at the N-terminus or C-terminus, or at both ends.
A protecting group covalently linked to the N-terminal amino group reduces the reactivity of the amino terminus under in vivo conditions. Amino protecting groups include -C1-10 alkyl, -C1-10 substituted alkyl, -C2-10 alkenyl, -C2-10 substituted alkenyl, aryl, -C1-6 alkylaryl, -C (O)-(CH2). 1-6-COOH, -C (O) -C1-6 alkyl, -C (O) -aryl, -C (O) -O-C1-6 alkyl, or -C (O) -O-aryl. It is done. Preferably, the amino terminal protecting group includes acetyl, propyl, succinyl, benzyl, benzyloxycarbonyl, or t-butyloxycarbonyl.

  A protecting group covalently linked to the C-terminal carboxy group reduces the reactivity of the carboxy terminus under in vivo conditions. The carboxy terminus protecting group is preferably attached to the a-carbonyl group of the final amino acid. Examples of the carboxy-terminal protecting group include amide, methylamide, and ethylamide.

Complex
PYY or a functional equivalent of PYY can be bound to other entities, for example to increase its half-life. The complex can improve delivery of the desired dose, inhibit degradation, and increase bioavailability in the circulation. A complex is any molecule.

  Complex preparation is well known in the art, e.g., Hermanson GT. Bioconjugate Techniques.New York: Academic Press; 1996, Aslam M, Dent AH. Bioconjugation: protein coupling techniques for the biomedical sciences.Houndsmills, England: See Macmillan Publishers; 1999, and Wong SS. Chemistry of protein conjugation and crosslinking. Boca Raton, FL: CRC Press; 1991.

  Most methods use amine reactive reagents or thiol reactive reagents. In preparing the conjugate, the use of certain linkers is advantageous. For example, linkers containing sterically-hindered disulfide bonds are often preferred because they are very stable in vivo and thus prevent release of toxic moieties prior to attachment at the site of action. It is generally desirable to have a complex that can be retained intact under conditions found everywhere in the body, other than the intended point of action where it is desired to exert good release.

Although different conjugates have been reported, for example, the use of the A chain of ricin is disclosed in US Pat. No. 4,340,535, which is hereby incorporated by reference. Examples of peptide conjugates based on Ac-RYY (RK) (WI) RK) -NH ₂ (parentheses indicate possible variants of amino acid residues) are disclosed in US Patent Application Publication No. 2003030472.

In one aspect of the invention, PYY or a functional equivalent thereof binds to another entity.
Before or after synthesis or purification, the molecules are linked as described above or by peptide bonds. The fusion can be obtained by any suitable method, including but not limited to DNA recombination methods. In a preferred embodiment, the fusion is by DNA recombination methods, for example, a fusion of a nucleotide sequence encoding a binding member and a nucleotide sequence encoding a ligand, whereby the fusion is encoded by a single nucleotide sequence. The fusion polypeptide is expressed and purified as a single polypeptide molecule using any suitable method described in Purification of binding members. The fusion polypeptide comprises an insertion of a linker such as, for example, a peptide of at least 2 AA, a peptide of at least 5 AA, a peptide of at least 8 AA, a peptide of at least 15 AA, a peptide of at least 20 AA.
In one aspect, PYY or a functional equivalent thereof binds to other entities using at least a 2AA linker.

PYY Preparation Method PYY or a functional equivalent thereof can be prepared using techniques well known in the art. For example, the polypeptide region of PYY can be synthesized and modified chemically or biochemically. Chemical synthesis techniques for prepeptides are well known in the art, for example solid phase peptide synthesis (see, eg, Vincent in Peptide and Protein Drug Delivery, New York, NY, Dekker, 1990). Examples of biochemical synthesis techniques including introduction of nucleic acids into cells and expression of nucleic acids are described in Ausubel, Current Protocols in Molecular Biology, John Willey, 1987-1998 and Sambrook et al., In Molecular Cloning, A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press, 1989.

  According to the present invention, PYY can be synthesized by solid phase peptide synthesis (see Example 1).

A second example of how PYY according to the present invention is prepared is shown briefly below. The human PYY of the present invention can be prepared by the following steps.
(A) constructing an expression vector containing an operon having a DNA sequence encoding human PYY or a functional equivalent thereof by a conventional method to prepare the vector of the present invention;
(B) Transfecting the expression vector into a host cell by a conventional method to prepare a transfected host cell of the present invention: (c) culturing the transfected cell by a conventional method and culturing the human PYY of the present invention To prepare.

  Host cells can be co-transfected with a second vector to optimize the manufacturing process. The two vectors contain different selectable markers. The coding sequence for PYY can contain cDNA or genomic DNA or both.

  The host cell used to express the PYY of the present invention may be, for example, a bacterial cell such as Escherichia Coli, or a eukaryotic cell such as S. cerevisiea or P. pastoris. In particular, mammalian cell lines are used, such as Hela, CHO, or any other suitable host cell known to those skilled in the art.

  The general methods for constructing the vectors of the invention, the transfection methods necessary to prepare the host cells of the invention, and the culture methods necessary to prepare the polypeptides of the invention from host cells are all conventional techniques. . Similarly, once prepared, the polypeptides of the invention are purified by the following standard steps.

After purification preparation of PYY, PYY or a functional equivalent thereof is preferably purified. The purification method used depends on several factors, for example, the purity required, the origin of PYY or functional equivalent thereof, the desired application, and the species from which PYY or functional equivalent is prepared.

  Any suitable routine method for polypeptide purification, such as precipitation or column chromatography, is well known to those skilled in the art of purification, for example, cross-flow filtration, ammonium sulfate fractionation, affinity column chromatography, gel Electrophoresis or the like is used.

PYY Composition According to the present invention, a composition containing PYY or a functional equivalent thereof is preferably prepared by chemical or biochemical synthesis or by recombinant methods, preferably present in blood such as infectious agents Does not contain any pollutants.
In preferred embodiments, the PYY composition is at least 10 nM, such as at least 50 nM, at least 0.2 μM, at least 0.5 μM, at least 1 μM, at least 2 μM, at least 5 μM, at least 10 μM, at least 20 μM, at least 50 μM, at least 0.2 mM, at least 0. Contains 5 μM, at least 1 mM, at least 2 mM, at least 5 mM PYY or a functional equivalent thereof.

  The PYY composition can be stored as a dry composition, for example, by freeze drying or spray drying to improve the stability of PYY. Such a composition is redissolved with a liquid solution before use. In general, the protein concentration of the reconstituted formulation is about 2-40 times greater than the protein concentration of the mixture prior to lyophilization, thus allowing the preparation of high concentration PYY compositions. When reconstituted with a diluent (such as sterile water for injection (BWFI)) containing a preservative, the reconstituted formulation can be used as a multi-dose formulation. Such formulations are useful, for example, when a patient requires frequent subcutaneous administration.

In other embodiments, the PYY composition is a highly stable liquid composition. The PYY composition is intended to be mixed with a suitable diluent prior to use.
The PYY composition may further comprise pharmacologically acceptable salts and pharmacologically acceptable carriers and diluents.

Pharmaceutical Compositions The pharmaceutical compositions of the present invention can be prepared by conventional methods such as those described in Remington: The Science and Practice of Pharmacy 1995, edited by EW Martin, Mack Publishing Company, 19th edition, Easton, Pa. The composition is in a conventional dosage form such as, for example, a solution or suspension.

  As used herein, the terms “pharmacologically acceptable”, “physiologically tolerable” and grammatical variations thereof, for example, when referring to a composition, carrier, diluent, reagent, etc. Used interchangeably, meaning that the substance can be administered to an individual without causing undesirable physiological effects such as nausea, dizziness, stomach discomfort, and the like.

According to the present invention, the pharmaceutical composition contains PYY or a functional equivalent thereof and a pharmacologically acceptable salt.
According to the invention, the pharmaceutical composition further preferably contains a pharmacologically acceptable salt, a pharmacologically acceptable carrier and / or diluent. The pharmaceutical composition can further contain vehicles, excipients, and / or transport molecules.
The pharmaceutical composition can be prepared prior to use by mixing the PYY composition with a suitable diluent.
The compositions of the present invention can be administered to an individual in any way that provides medicinal effects, preferably for the treatment of gastrointestinal diseases.

Aspects of the present invention relate to a method of treatment characterized by administering a pharmaceutical composition according to the present invention.
The pharmaceutical composition according to the present invention is formulated to be suitable for administration by any suitable method such as peripheral administration, parenteral administration or oral administration.
The pharmaceutical composition of the present invention is preferably suitable for parenteral administration, for example subcutaneous, intramuscular, intravenous, intranasal, inhalation, buccal, intradermal and transdermal administration, and also by rectal suppository. Formulated into a formulation.

Additional treatment is effective for patients suffering from a second active ingredient eating disorder event. This includes, for example, selective serotonin reuptake inhibitors (SSRI), serotonin / noradrenaline reuptake inhibitors (SNRI), norepinephrine / serotonin reuptake inhibitors (NSRI), selective noradrenaline reuptake inhibitors, tetracyclic anticancer drugs Non-selective monoamine reuptake inhibitors such as antidepressants, tricyclic antidepressants (TCAs), selective reversible monoamine reuptake inhibitors, and antidepressants with other mechanisms of action, mirtazapine, etc. An antidepressant is mentioned. Examples of SSRIs are citalopram, escitalopram, fluoxetine, fluvoxamine, paroxetine and sertraline. An example of SNRI is venlafaxine. NSRI includes the Milinashi plan.

In an embodiment of the invention, the composition can contain a second active ingredient in addition to PYY or a functional equivalent thereof.
In other embodiments, antiemetics can be used as the second active ingredient. This is particularly useful when nausea and / or vomiting is caused by treatment with PYY or a functional equivalent thereof.

Antiemetics In this context, antiemetics are those that counteract (reduce or eliminate) nausea and / or vomiting. Nausea and vomiting experiences have many causes, and treatment or reduction of this symptom can be achieved by a variety of mechanisms. The main groups of drugs useful for the treatment of nausea and vomiting are: neurostabilizers / antipsychotics, antihistamines, anticholinergics, steroids (corticosteroids), 5-HT3 receptor antagonists (serotonin receptor antagonists) NK1 receptor antagonist (neurokinin 1 substance P receptor antagonist), anti-dopaminergic agent / dopamine receptor antagonist, benzodiazepine, cannabinoid. Below is a non-limiting list of each compound in a different group.
1. Nerve stabilizer / antipsychotic a. Dixyrazine
b. Haloperidol
c. Prochlorperazine (Compazine registered trademark)
2. Antihistamine a. Piperazine derivatives
i. Cyclidine
ii. Meclizine
iii. Cinnarizine b. Promethazine c. Dimenhydrinate d. Diphenhydramine e. Hydroxyzine f. Buclizine
g. Meclizine hydrochloride (Bonine, Antivert)
3. Anticholinergic agents (acetylcholine receptor inhibitors)
a. Scopolamine b. Glycopyrron
c. Hyoscine i. Artan (trihexyl-5-trihexylphenidyl hydrochloride)
ii. Kogentin (benztropine mesylate)
iii. Aquinetone (biperiden hydrochloride)
iv. DISIPAL (Norflex Citrate Orphenadrine)
v. Chemadrine (Procyclidine hydrochloride)
4). Steroids (corticosteroids)
a. Betamethasone b. Dexamethasone c. Methylprednisolone d. Prednisone (registered trademark)
e. Trimethobenzamide (Tigan)
5.5-HT3 receptor antagonist (serotonin receptor antagonist)
a. Granisetron b. Dorasetron c. Ondansetron (hydrochloride)
d. Tropisetron
e. Ramosetron f. Palonosetron g. Arosetron h. Bemesetron i. Zachisetron j. Batanopilde k. MDL-73147EF
l. Metoclopramide m. N-3389 (endo-3,9-dimethyl-3,9-diazabicyclo [3.3.1] non-7-yl-1H-indazole-3-carbosamide dihydrochloride)
n. Y-25130 hydrochloride o. MDL 72222
p. Tropanyl-3,5-dimethylbenzoate q. 3- (4-allylpiperazin-1-yl) -2-quinoxaline carbonyl maleate r. Zacoprid hydrochloride s. Mirtazapine (Antidepressant)
6). NK1 receptor antagonist (neurokinin 1 substance P receptor antagonist)
a. Aprepitant b. MPC-4505
c. GW597599
d. MPC-4505
e. GR205171 (selective tachykinin NK1 receptor antagonist)
f. L-759274
g. SR 140333
h. CP-96,345
i. BIIF 1149, NKP 608C, NKP 608A, CGP 60829, SR 140333 (Nolpitantium besilate / chloride), LY 303870 (Lanepitant), MDL-105172A, MDL-103896, MEN-11149, MEN-11467, DNK 333A, YM-49244, YM-44778, ZM-274773, MEM-10930, S-19752, Neuronorm, YM-35375, DA-5018, MK-869, L-754030, CJ-11974, L-758298, DNK-33A, 6b-I, CJ-11974
j. Benserazide and carbidopa k. TAK-637 [(aR, 9R) -7- [3,5-bis (trifluoromethyl) benzyl] -8,9,10,11-tetrahydro-9-methyl-5- (4-methylphenyl) -7H -[1,4] diazocino [2,1-g] [1,7] naphthyridine-6,13-dione]
l. PD 154075 ([(2-Benzofuran) -CH2OCO]-(R) -α-MeTrp- (S) -NHCH (CH3) Ph)
m. FK888, chemical modification of parent compound, (D-Pro4, D-Typ7,9,10, Phe11) SP4-11
7). Anti-dopamine agonist / dopamine receptor antagonist a. Domperidone b. Prochloroperazine c. Metoclopramide d. Chlorpromazine (Thorazine)
e. Droperidol f. Promethazine (Phenergan)
8). Benzodiazepine (Barium (R) etc.)
9. Non-neuroactive cannabinoid a. Cannabidiol (CBD)
b. Dimethylheptylcannabidiol (CBD-DMH)
c. Tetrahydrocannabinol (THC)
d. Cannabinoid agonists such as WIN55-212 (CB1 and CB2 receptor agonists)
e. Dronabinol (Marinol (registered trademark))
10. Additional cannabinoids a. Naviron (Cesamet)
11. c-9280 (Merck)

  A 5-HT3 receptor antagonist is particularly preferred. Antiemetics are used in an effective amount sufficient to eliminate nausea and / or vomiting or to reduce to an acceptable level.

Preferred embodiments The pharmaceutical composition comprises a selective serotonin reuptake inhibitor (SNRI), a serotonin / noradrenaline reuptake inhibitor (SNRI), a norepinephrine / serotonin reuptake inhibitor (NSRI), a selective noradrenaline reuptake inhibitor, a tetracycle Non-selective monoamine reuptake inhibitors, including systemic antidepressants, tricyclic antidepressants (TCAs), selective reversible monoamine reuptake inhibitors, and antidepressants with other mechanisms of action, such as A kit-in-part preparation further containing an antidepressant such as mirtazapine. Examples of SSRIs include citalopram, escitalopram, fluoxetine, fluvoxamine, paroxetine and sertraline. An example of SNRI is venlafaxine. This kit-in-part formulation can be used for simultaneous, sequential or separate administration. An example of NSRI is the milnacipran.
In a preferred embodiment, the pharmaceutical composition is a kit-in-part formulation.

Pharmacologically acceptable salts Pharmacologically acceptable salts of the compounds of the invention are also intended to be included within the scope of the invention if they can be prepared. These salts refer to those that are acceptable for pharmaceutical use. That is, the salt means one that retains the biological activity of the parent compound and does not exhibit deleterious or exacerbating effects when administered and used in the treatment of disease.

Pharmacologically acceptable salts can be prepared by standard techniques. When the parent compound is a base, it is treated with an excess of organic or inorganic acid in a suitable solvent. When the parent compound is an acid, it is treated with an inorganic or organic base in a suitable solvent.
The compound of the present invention is in the form of its alkali metal or alkaline earth metal salt, particularly and preferably in the form of a pharmaceutical composition, together with, or simultaneously with, a pharmaceutically acceptable carrier or diluent. An effective amount can be administered by oral, rectal, or parenteral route (including subcutaneous administration).

  Examples of pharmaceutically acceptable acid addition salts used in the pharmaceutical composition of the present invention include those derived from inorganic acids (hydrochloric acid, bromic acid, phosphoric acid, metaphosphoric acid, nitric acid and sulfuric acid), organic acids (tartaric acid, acetic acid) , Citric acid, malic acid, lactic acid, fumaric acid, benzoic acid, glycolic acid, glucuronic acid, succinic acid, p-toluenesulfonic acid, arylsulfonic acid).

  The pharmaceutical compositions of the invention may include pharmacologically acceptable salts of the compounds disclosed herein. Pharmacologically acceptable salts include acid addition salts (formed with the free amino group of the polypeptide).

  Such salts include pharmacologically acceptable acid addition salts, pharmacologically acceptable metal salts, ammonium salts and alkylated ammonium salts. Examples of acid addition salts include salts with inorganic acids and salts with organic acids. Representative examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, sulfuric acid, nitric acid and the like. Representative examples of suitable organic acids include formic acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, propionic acid, benzoic acid, cinnamic acid, citric acid, fumaric acid, glycolic acid, lactic acid, maleic acid, malic acid, malonic acid, Mandelic acid, oxalic acid, picric acid, pyruvic acid, salicylic acid, succinic acid, methanesulfonic acid, ethanesulfonic acid, tartaric acid, ascorbic acid, pamonic acid, bismethylenesalicylic acid, ethanedisulfonic acid, gluconic acid, citraconic acid, aspartic acid, Examples include stearic acid, palmitic acid, ethylenediaminetetraacetic acid (EDTA), p-aminobenzoic acid, glutamic acid, benzenesulfonic acid, and p-toluenesulfonic acid. Further examples of pharmacologically acceptable inorganic or organic acid addition salts include the pharmacologically acceptable salts listed in J. Pharm. Sci. 1977, 66.2, which is incorporated by reference. Included in this specification. Examples of metal salts include lithium, sodium, potassium, and magnesium salts.

  According to the invention, organic acid salts of organic acids such as acetic acid are preferred.

  Examples of ammonium and alkylated ammonium salts include ammonium, methylammonium, dimethylammonium, trimethylammonium, ethylammonium, hydroxyethylammonium, diethylammonium, butylammonium, and tetramethylammonium salts.

Salts formed with free carboxyl groups are, for example, those derived from inorganic bases (sodium, potassium, ammonium, calcium hydroxide or ferric hydroxide, etc.) and organic bases (isopropylamine, trimethylamine, 2-ethylamino) Ethanol, histidine, procaine, etc.) may be used.
Moreover, those arbitrary hydrates (hydrated form) are also included in the scope of the compounds of the present invention and pharmacologically acceptable acid addition salts thereof.

  The preparation of a pharmaceutical composition in which an active ingredient is dissolved or dispersed is well known in the art. While such compositions are primarily prepared as liquid solutions or suspensions, aqueous or non-aqueous sterile injections, solid forms suitable for solutions or suspensions in liquid can also be prepared prior to use. The preparation may be emulsified.

The pharmaceutically acceptable carrier and diluent active ingredients can be mixed with excipients that are pharmaceutically acceptable and compatible with the active ingredients in amounts suitable for use in the therapeutic methods disclosed herein. Suitable excipients include, for example, water, saline, dextrose, glycerol, ethanol and the like or combinations thereof. Furthermore, if necessary, the composition may contain small amounts of adjuvants (those that increase the effect of active ingredients such as wetting agents, emulsifying agents, pH buffering agents).

  The liquid composition may also contain a liquid phase other than water or excluding water. Examples of such additional liquid phases include glycerin, vegetable oils (such as cottonseed oil), organic esters (such as ethyl oleate), and water oil emulsions.

  Suitable pharmaceutically acceptable carriers include inert solid diluents and fillers, sterile aqueous solutions and various organic solvents. Examples of solid carriers include lactose, gypsum, sucrose, cyclodextrin, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid or lower alkyl ethers of cellulose. Examples of liquid carriers include syrup, peanut oil, olive oil, phospholipid, fatty acid, fatty acid amine, polyoxyethylene or water.

  A pharmaceutical composition formed by mixing a compound of the present invention and a pharmacologically acceptable carrier can be readily administered in a variety of dosage forms suitable for the disclosed route of administration. The composition is advantageously formulated into single dose or multiple dose forms by methods known in the pharmaceutical arts.

  In a further aspect, the invention relates to a pharmaceutical composition comprising as an active ingredient a compound as described above or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

Stabilizers The active compounds of the present invention may be unstable, so the composition may preferably contain stabilizers, preservatives, or preservatives to increase the stability of the compound.

  A pH buffering agent is used to stabilize the active compound of the composition. The buffer is acetate, carboxylate, dicarboxylate, phosphate, citrate, Tris or Hepes. In a preferred embodiment, the buffering agent is acetate.

  According to the present invention, the composition is preferably from 2.0 to 9.0, alternatively from 2.5 to 8.0, alternatively from 3.0 to 7.0, alternatively from 3.5 to 6.0, alternatively 3.5. PH value between 5.0 and 5.0, alternatively 4.0 to 5.5, alternatively 4.0 to 5.0, alternatively 4.0 to 4.5. Preferably, the pH of the composition is 6 or less, preferably 5.5 or less, preferably 5 or less, preferably 4.8 or less, preferably 4.6 or less, preferably 4.4 or less, preferably 4.2. It is as follows.

Tween 20, tween 60, tween 80, span 80, cetostearyl alcohol, benzyl alcohol, myristyl alcohol, glyceryl monostearate, mannitol, polysorbate and sodium lauryl sulfate are possible stabilizers.
In a preferred embodiment, mannitol is used as a stabilizer.

Tween 60, span 80, cetostearyl alcohol, benzyl alcohol, myristyl alcohol, glyceryl monostearate, mannitol and sodium lauryl sulfate are possible stabilizers.
In a preferred embodiment, mannitol is used as a stabilizer.

In preparing lyophilized compositions, lyoprotectants are used to stabilize the active ingredient (Twonsend and DeLuca, "Use of lyoprotectants in the freez-drying of a model protein, ribonuclease A" Journal of Parenteral Science & Technology 42 (6): 190-199 (Nov.-Dec. 1988)).
The dispersion protecting agent is preferably sucrose such as sucrose, dextran, or hydroxypropyl- / 142-cyclodextrin, or a sugar such as trehalose.

Transport molecules Transport molecules act by incorporating or immobilizing the compounds of the present invention therein. Any suitable transport molecule known to the technician can be used. Examples of transport molecules include liposomes, micelles, and / or microspheres.
For example, as described in Szoka et al., Ann. Rev. Biophys. Bioeng. 9: 467 (1980), U.S. Pat. Nos. 4,235,871, 4501728, 4837028, all of which are incorporated herein by reference. Can be used to prepare liposomes.
Micelles are formed by surfactants (molecules having a hydrophobic portion and one or more ionic or strongly hydrophilic groups) in an aqueous solution. As the concentration of the solid surfactant increases, the monolayer adsorbed on the air / water or glass / water interface becomes more compact and further occupancy is already in the two monolayers. Requires excessive compression of existing interfacial molecules. Further increase in the amount of surfactant dissolved above that concentration yields the same amount as the new molecule and aggregates in the micelle. This process starts with a characteristic concentration called “critical micelle concentration”.

  The shape of the micelle formed in the diluted surfactant solution is approximately spherical. The polar head group of the surfactant molecule is placed in the outer spherical shell with the hydrocarbon chain oriented in the center, forming the spherical nucleus of the micelle. The hydrocarbon chains are arbitrarily coiled and the micelle interior is monopolar and liquid. In micelles of polyoxyethylenated nonionic surfactant, the polyoxyethylene sites are oriented outward and penetrated into water. This arrangement is energetically desirable because the hydrophilic head group is in contact with water and the carbon hydrogen moiety is removed from the aqueous medium and partially hidden from contact with water by the polar head group. The hydrocarbon tails of the surfactant molecules are located inside the micelles and interact with each other with a weak van der Waals force.

  The size of micelles or their aggregation number is mainly governed by geometrical factors. The radius of the carbon hydrogen nucleus cannot exceed the length of the extended hydrocarbon chain of the surfactant molecule. Therefore, the aggregation number of spherical micelles is increased by increasing the chain length or increasing the number of homologous columns. If the surfactant concentration increases by more than a few percent, and if an electrolyte is added (for ionic surfactants) or if the temperature is increased (for nonionic surfactants), the micelles Increase its size. Under these conditions, the micelles are too large to remain spherical, and become elliptical, cylindrical, or fine layered.

  Common surfactants well known to those skilled in the art are used in the micelles of the present invention. Suitable surfactants include sodium laurate, sodium oleate, sodium lauryl sulfate, octaoxyethylene glycol monododecyl ether, octoxynol 9 and Pluronic F-127 (PLURONIC F-127, Wyandotte Chemicals Corp.). It is done. Suitable surfactants are nonionic polyoxyethylene and polyoxypropylene surfactants conforming to intravenous injection, such as Tween 80, Pluronic F-68, n-octyl-beta-D-glucopyranosyl and the like. Furthermore, the phospholipids described in the use in preparing liposomes can also be used for micelle formation.

Compositions for oral administration The compounds of the present invention are formulated into a wide range of dosage forms for oral administration. Pharmaceutical compositions and dosage forms comprise the compound of the present invention or a pharmacologically acceptable salt thereof or a crystal form thereof as an active compound. The pharmacologically acceptable carrier may be solid or liquid. Solid dosage forms include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. Solid carriers include one or more substances that act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, humidifiers, tablet disintegrating agents, or encapsulating agents.
Preferably, the balance consisting of suitable pharmacological excipients is from 0.00002% to 2% by weight of the compound of the invention. Such excipients for oral administration include pharmaceutical grade mannitol, lactose, starch, magnesium stearate, sodium saccharine, talc, cellulose, glucose, gelatin, sucrose, magnesium carbonate and the like.

  In powders, the carrier is a finely divided solid which is a mixture with the finely divided active component. In tablets, the active ingredient is mixed with the carrier having the necessary binding capacity in suitable proportions and compressed into the required shape and size. Powders and tablets preferably contain 1-70% active ingredient. Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like. The term “preparation” also encompasses the preparation of encapsulant and active compound as a carrier to provide a capsule in which the active ingredient is surrounded by the carrier associated with it, with or without other carriers. And Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges are solid preparations for oral administration.

  The drops of the invention are sterile or non-sterile aqueous or oily solutions or suspensions, sometimes with an active ingredient, sometimes a bactericidal and / or antifungal agent and / or any suitable preservative, and sometimes a surfactant. It can be prepared by dissolving in a suitable aqueous solution containing. The resulting solution is filtered and clarified, transferred to a suitable container, then sealed and autoclaved or sterilized by heating to 98-100 ° C for half an hour. Alternatively, the solution is sterilized by filtration and transferred aseptically to a container. Examples of fungicides and antifungal agents suitable for inclusion in drops include phenylmercuric nitrate or phenylmercuric acetate (0.002%), benzalkonium chloride (0.01%) and chlorohexidine acetate (0. 01%). Suitable solvents for the preparation of oily solutions are glycerol, dilute alcohol and propylene alcohol.

  Also included are solid formulations that are intended to be converted to liquid formulations for oral administration just prior to use. Such liquid dosage forms include solutions, suspensions, and emulsions. Such formulations may contain, in addition to the active compound, colorants, flavors, stabilizers, buffers, artificial or natural sweeteners, dispersants, thickeners, solubilizer adjuvants, and the like.

  Other dosage forms suitable for oral administration include emulsions, syrups, elixirs, aqueous solutions, aqueous suspensions, toothpastes, gel dentifrices, chewing gum and other liquid formulations, or are intended to be changed to liquid formulations immediately before use. Solid formulations prepared. The emulsion may be prepared in a solution in a water-soluble propylene glycol solution or may contain an emulsifier (such as lecithin, sorbitan monooleate, or acacia). Aqueous solutions can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizers, and thickening agents. Water-soluble suspensions are prepared by dispersing finely divided active ingredients in water together with sticky substances (natural or synthetic rubbers, resins, methylcellulose, sodium carboxymethylcellulose, and other well-known suspending agents). I can do it. Solid preparations include solutions, suspensions, and emulsions. In addition to the active ingredient, coloring agents, flavoring agents, stabilizers, buffering agents, artificial or natural sweeteners, dispersing agents, thickeners, solubilizing aids An agent or the like may be contained.

A composition for parenteral administration can take the form of a suspension, solution, or oily or aqueous medium emulsion, such as a solution in aqueous polyethylene glycol. Examples of oily or non-aqueous carriers, diluents, solvents or media include propylene glycol, polyethylene glycol, vegetable oils (such as olive oil), and injectable organic esters (such as oleates), as well as formulation agents, For example, preservatives, wetting agents, emulsifying agents, suspending agents, stabilizers and / or dispersing agents may be included. Alternatively, the active ingredient may be in the form of a powder obtained by aseptic extraction of a sterilized solid or lyophilization of a solution for reconstitution prior to use in a suitable medium (such as sterile, pyrogen-free water). Also good. If necessary, the aqueous solution should preferably be buffered, and the liquid diluent is first made isotonic with sufficient saline or glucose. Aqueous solutions are particularly suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. All sterile aqueous media used are readily obtained by standard techniques known to those skilled in the art.

  In a preferred embodiment of the present invention, the composition of PYY or a functional equivalent thereof or a salt thereof is a lyophilized composition, and may further contain a solvent. In other embodiments, the composition is a solution of PYY or a functional equivalent or salt thereof according to the present invention. Preferably, the solvent may be any suitable solvent described herein, and in particular the solvent is a saline solution or a physiological buffer such as a phosphate buffer.

The pharmaceutical composition contains PYY or a functional equivalent or pharmacologically acceptable salt thereof (and, for example, antigenic epitopes and protease inhibitors). Such compositions are prepared by mixing with water or saline, and sometimes with a non-toxic surfactant. Compositions for intravenous or intraarterial administration are sterile aqueous solutions containing buffers, liposomes, diluents and other suitable additives.
Oils used in parenteral compositions include petroleum, animal, vegetable or synthetic oils. Specific examples of oils useful in such compositions include peanut oil, soybean oil, sesame oil, cottonseed oil, corn oil, olive oil, liquid paraffin, mineral oil. Suitable fatty acids for use in the parenteral composition include oleic acid, stearic acid, and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters.

  Soaps used in parenteral compositions include fatty acid alkali metal salts, ammonium salts, and triethanolamine salts, and suitable surfactants include (a) dimethyldialkylammonium halides. Cationic surfactants such as alkylpyridinium halides, (b) anionic surfactants such as alkyl sulfonates, aryl and olefins, alkyl sulfates, olefins, ethers, monoglycerides, and sulfosuccinates, (c) fatty acid amine oxides, Nonionic surfactants such as fatty acid alkanolamides, polyoxyethylene polypropylene copolymers, (d) amphoteric surfactants such as (d) alkyl-beta-aminopropionate and 2-alkyl-imidazoline quaternary ammonium salts, and (d )these A mixture is mentioned.

In general, compositions for parenteral administration contain from about 0.5 to about 25% (wt%) active ingredient in solution. Preservatives and buffers can be used. In order to minimize or eliminate irritation at the injection site, such a composition is one of the nonionic surfactants having a hydrophilic-lipophilic balance (HLB) of about 12 to about 17. Or you may contain multiple. The amount of surfactant in such compositions is usually in the range of about 5 to about 15% by weight. Suitable surfactants include polyethylene sorbitan fatty acid esters (such as sorbitan monooleate) and polymer adducts of ethylene oxide with a hydrophobic base obtained by condensation of propylene glycol and propylene oxide. The parenteral composition is stored in a single-dose or multiple-dose sealed container (ampoules or vials), stored under lyophilized (freeze-dried) conditions, and a sterile liquid vehicle for injection just before use. For example, only water is required. Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules, and tablets described above.
Injectable pharmaceutical dosage forms include sterile aqueous solutions or dispersions containing the active ingredients which are adapted for administration by encapsulation in liposomes. In all cases, the ultimate dosage form must be sterile, fluid and stable under the conditions of the manufacturing and storage process.

  By mixing the required amount of PYY or a functional equivalent thereof or a pharmaceutically acceptable salt thereof together with some of the other ingredients listed above in a suitable solvent and then filter sterilizing if necessary. Sterile injectable solutions can be prepared.

Compositions for topical administration The compositions of the present invention can be delivered topically. Sites for topical administration include mucosal tissues of the epidermis and rectum, nose, mouth and throat. Compositions for topical administration via the skin and mucous membranes should not cause irritation aspects such as swelling and redness.
Topical compositions include a pharmaceutically acceptable carrier adapted for topical administration. Therefore, the composition can be a suspension, solution, ointment, lotion, claim, foam, aerosol, spray, suppository, embedding, inhalant, tablet, capsule, dry powder, syrup, balm or lozenge (troche), etc. Can take the form of Methods for preparing such compositions are well known in the pharmaceutical industry.

  The compositions of the invention are formulated as ointments, claims or lotions, or transdermal patches for topical administration to the epidermis. Ointments and claims can be formulated, for example, by the addition of an aqueous or oily base and a suitable thickening and / or gelling agent. Lotions are formulated with an aqueous or oily base and will usually also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents, thickening agents, coloring agents. A composition suitable for topical administration in the mouth is a medicated candy containing a base flavored with an active ingredient, usually sucrose, acacia or tracant; an active ingredient with an inert base (gelatin, glycerin, A sucrose, acacia) aromatic tablet (pastille); a mouthwash containing the active ingredient in a suitable liquid carrier.

  The cream, ointment or paste of the present invention is a semi-solid composition for external use of the active ingredient. They can be used as a fatty base or non-fatty base by using a suitable machine for the active ingredient finely decomposed and pulverized alone, in a solution or suspension in an aqueous or non-aqueous fluid. It can be prepared by mixing with. Bases are hard, soft, or liquid hydrocarbons such as paraffin, glycerol, beeswax, metal soap; rubber paste; natural oils such as almond oil, corn oil, peanut oil, castor oil, olive oil; wool Examples thereof include fats or derivatives thereof, fatty acids such as stearic acid and oleic acid, and alcohols such as propylene glycol and macrogol. The composition can incorporate a suitable surfactant, such as an anionic, cationic, or nonionic surfactant, such as a sorbitan ester or its polyoxyethylene derivative. Suspending agents such as natural gums, cellulose derivatives and inorganic materials such as silica and other ingredients such as lanolin can also be included.

  The lotion of the present invention is suitable for administration to the skin and eyes. The eye drops are bactericidal aqueous solutions that may contain bactericides and can be prepared in a similar manner to the preparation of drops. Lotions or liniments to be administered to the skin also include agents that accelerate drying such as alcohol or acetone and cool the skin, and / or humectants such as oils such as glycerol, castor oil and peanut oil.

  The pharmaceutical-chemical modifier complex described herein can be administered transdermally. Transdermal administration usually means drug delivery by a transdermal route to the systemic circulation of the patient. The skin site means an anatomical region for transdermal administration of a drug, and examples include the upper arm, abdomen, chest, back, buttocks, and nipple.

  Transdermal delivery is accomplished by prolonged exposure of the source of the complex to the patient's skin. Transdermal patches have the added advantage of providing controlled delivery of the drug-chemical modifier complex to the body. Transdermal Drug Deliveary, Developmental Issues and Research Initiatives, Hadgraft and Guy (eds.), Marcel Dekker, Inc., (1989); Controlled Drug Delivery: Fundamentals and Applications, Robinson and Lee (eds.), See Marcel Dekker Inc., (1987); and Transdermal Delivery of Drugs, Vols. 1-3, Kydonieous and Berner (eds.), CRC Press, (1987). Such dosage formulations can be prepared by dissolving, dispersing or incorporating the pharmaceutical-chemical modifier complex in a suitable medium, such as an elastomeric material. Absorption enhancers are also used to increase the flux of the compound across the skin. Such a flow rate can be adjusted by providing a rate-limiting membrane or by dispersing the compound in a polymer matrix or gel.

  Various types of transdermal patches can be used in the methods disclosed herein. For example, a simple adsorption patch can be prepared from a substrate and an acrylic adhesive. The drug-chemical modifier complex and optional enhancer are molded into the adsorption template solution and thoroughly mixed. The solution is poured directly onto the substrate, and the casting solvent is distilled off in an oven to obtain an adhesive film. The release liner is glued to complete the system.

  Alternatively, polurethane matrix patches can be used to deliver drug-chemical modifier complexes. This patch layer consists of a substrate, a polyurethane agent / enhancer matrix, a membrane, an adhesive, and a release liner. The polyurethane matrix is prepared using a room temperature curing polyurethane prepolymer. By adding water, alcohol, and complex to the prepolymer, a sticky film-like elastomer is formed, which is cast directly onto the substrate.

  In a further aspect of the invention, a hydrogel matrix patch is used. In general, the hydrogel matrix consists of alcohol, water, drug, and several hydrophilic polymers. This hydrogel matrix is incorporated into a transdermal patch between the substrate and the adhesive layer.

  Liquid reservoir patches can also be used in the methods disclosed herein. The patch consists of an impermeable or semi-permeable, heat-sealable substrate, a heat-sealable membrane, a skin adhesive that can be bonded only by acrylate pressure, and a siliconized release liner. The substrate is heat sealed to the membrane to form a reservoir that is then filled with a solution of the complex, enhancer, gelling agent, and other excipients.

  Foam matrix patches are similar in design and ingredients to liquid reservoir patches, except that the gelled pharmaceutical-chemical modifier solution is confined to a thin foam layer, usually polyurethane. This foam layer is located between the base material and the membrane heat sealed at the outer periphery of the patch.

  For passive delivery systems, the release rate is generally controlled by diffusion from a monolithic device with a membrane located between the reservoir and the skin, or by the skin itself, which acts as a rate-limiting barrier in the delivery system. See U.S. Pat. Nos. 4,816,258, 4927405, 4904475, 4588580, 488062 and the like. The drug delivery rate depends in part on the nature of the membrane. For example, transmembrane drug delivery rates in the body are generally faster than when permeating the skin barrier. The rate at which the complex is delivered from the device to the membrane can be most advantageously adjusted by the use of a rate limiting membrane located between the reservoir and the skin. Assuming that the skin is sufficiently permeable to the complex (eg, absorption through the skin is greater than the speed of the route through the membrane), the membrane serves to regulate the rate at which the patient is obtained Will.

  Suitable permeable membrane materials are selected based on the desired permeability, composite properties, and mechanical conditions regarding the device configuration. Specific examples of permeable membrane materials include a wide range of natural and synthetic polymers such as polydimethylsiloxane (silicone rubber), ethylene vinyl acetate copolymer (EVA), polyurethane, polyurethane-polyether copolymer, polyurethane, polyamide, polyvinyl chloride (PVC). ), Polypropylene, polycarbonate, polytetrafluoroethylene (PTEE), cellulosic materials such as cellulose triacetate and cellulose nitrate / acetate, and hydrogels such as 2-hydroxyethyl methacrylate (HEMA).

  The device can also contain other items, such as other conventional components of therapeutic substances, depending on the nature of the desired device. For example, the compositions of the present invention can include one or more preservatives or fungicides, such as methyl hydroxybenzoate, propyl hydroxybenzoate, chlorocresol, benzalkonium chloride, and the like. These pharmaceutical compositions may also contain other active ingredients such as antibacterial agents, in particular antibiotics, anesthetics, analgesics, and antipruritic agents.

Compositions to be Administered as Suppositories The compounds of the invention can be formulated for administration as suppositories. A low melting wax such as a mixture of fatty acid glycerides or cocoa butter is first melted, and the active ingredient is uniformly dispersed by, for example, stirring. The molten homogeneous mixture is then poured into a normal size mold, allowed to cool and solidify.
The active compound is formed into a suppository, eg, containing about 0.5% to about 50% of a compound of the invention in a polyethylene glycol (PEG) carrier (eg, PEG 1000 [96%] and PEG 4000 [4%]) .

Composition for nasal administration The compound of the present invention is formulated for nasal administration. Solution or suspension formulations are administered directly into the nasal cavity by conventional means, for example with a dropper, pipette or spray. The composition is provided in single or multiple dose forms. In the latter case of a dropper or pipette, it is performed by the patient himself by administering a suitable predetermined amount of a solution or suspension formulation. In the case of spraying, for example, a metering spray pump is used.

Composition for aerosol administration The compounds of the present invention can be formulated for aerosol administration, particularly for the respiratory tract, including intranasal administration. The compound generally has a small particle size of, for example, a unit of 5 microns or less. Such particle size is obtained by methods known in the art, such as micronization. The active ingredient is pressurized with a suitable high-pressure gas (propellant) such as, for example, chlorofluorocarbon (CFC, eg dichlorodifluoromethane, trichlorofluoromethane) or dichlorotetrafluoroethane), carbon dioxide, or other suitable gas. Offered in packs. Aerosols usually also contain a surfactant such as lecithin. The drug dosage is adjusted by a metering valve. Alternatively, the active ingredient is provided in a dry powder dosage form, such as a powder mixture of suitable powders and compounds, such as lactose, starch, starch derivatives such as hydroxypropylmethylcellulose and polyvinylpyrrolidine (PVP), etc. Served in dry powder. The powder carrier is gelled in the nasal cavity. Powder compositions are provided in unit dosage forms, such as gelatin capsules or cartridges, or blister packs in which the powder can be administered by inhaler.

  Compositions administered by aerosol include saline solutions using, for example, benzyl alcohol and other suitable preservatives, absorption enhancers that promote bioavailability, fluorocarbons, and / or other solubilizers or dispersants Can be prepared.

The composition of the mixture that the present invention may be administered in combination with a second active ingredient, is further envisioned. The second active ingredient is formulated with other ingredients of the present invention.
It is further envisioned that the active ingredient of the present invention can be administered in combination with a second active ingredient. “In combination” means that the composition is formulated with other compounds in the same composition, and / or the second active ingredient and / or pharmaceutical composition is the present invention. Is administered before, during (including simultaneously) and / or after administration of the composition.
Accordingly, one aspect of the present invention relates to a composition of PYY or a functional equivalent thereof and a second active ingredient.
Thus, one aspect of the present invention relates to a composition of PYY or a functional equivalent thereof described herein and a second active ingredient. The second active ingredient includes, but is not limited to, beta adrenergic receptor antagonists, alpha adrenergic receptor antagonists, calcium antagonists, angiotensin converting enzyme (ACE) inhibitors, angiotensin II receptor antagonists and diuretics Antihypertensive agents such as (but not limited to, thiazide diuretics and loop diuretics); insulin; DPP-V inhibitors; but not limited to selective serotonin reuptake inhibitors (SSRIs), mianserin and mirtazapine, Antidepressants such as classic and atypical psychotropic drugs (nerve stabilizers); corticosteroids; and other drugs that increase body weight or body fat, or high blood pressure, overweight, obesity, metabolic syndrome (syndrome X) And / or other drugs that are intended for the treatment of diabetes.

Administration Further within the scope of the present invention is a method of treating an individual in need, characterized by administering to the individual one or more effective doses of the pharmaceutical composition disclosed herein. The individual is preferably one or more of the health problems mentioned above, such as those suffering from, or at risk of suffering from gastrointestinal diseases. “Treatment” also means prevention and aftercare, and / or means the prevention and / or treatment of a disease that reduces and / or reduces pathological symptoms. The methods of treatment include improving an individual's well-being awareness and quality of life. The method refers to one or more combination therapies disclosed herein.

  The pharmaceutical composition of the present invention can be used for both prophylactic and therapeutic administration. Thus, a pharmaceutical composition of PYY or a functional equivalent thereof can be administered to a patient to minimize the severity of the disease, to prevent worsening of gastrointestinal disease, or to a patient already suffering from the disease. Can be administered. Furthermore, the treatment method prevents recurrence of gastrointestinal disease.

The pharmaceutical composition can be prepared to suit one or more specific methods of administration. The compositions of the invention can be administered parenterally, orally, nasally (inhalation or nasal spray), topically (in the skin or eye), rectally using suppositories, or by vaginal absorption. it can.
A pharmaceutical composition of PYY or a functional equivalent thereof can be administered to an individual in need.
According to the invention, the composition of PYY or a functional equivalent thereof can be administered parenterally, for example by injection. Thus, according to the present invention, PYY or a functional equivalent thereof can be administered parenterally, such as intravenously, intraarterially, intraperitoneally, intramuscularly, subcutaneously, intranasally, or transdermally. The composition can be further administered by inhalation, topical administration to the eye, nasal administration, vaginal absorption, dermal administration, and rectal suppository. Other methods of drug administration that are effective to deliver the drug to the target site or introduce the drug into the bloodstream are also contemplated.

  A pharmaceutical composition of PYY or a functional equivalent thereof can be administered intravenously, eg, by unit dose injection. The term “unit dose” as used in connection with the pharmaceutical composition of the present invention means a physically discrete unit suitable as a single dose for a subject, each unit being calculated to obtain the desired therapeutic effect. A predetermined amount of the active substance contained is contained together with the required diluent (eg carrier or medium). Alternatively, the pharmaceutical composition can be prepared in multiple dosage forms, each single dosage form being administered as needed using a multiple dose delivery device. As an alternative to standard injection, the pharmaceutical composition can be administered by infusion (such infusion is preferably short-lived).

  According to the present invention, it is believed that the effects of PYY or its functional equivalent are mediated by the action of PYY outside the central nervous system, except for NPY neurons in the hypothalamus. Thus, PYY or its functional equivalent does not affect NPY neurons located throughout the central nervous system.

Thus, PYY or a functional equivalent thereof that can bind to the NPY Y2 receptor circulates in the bloodstream to the hypothalamic receptor. However, these molecules preferably cannot cross the blood brain barrier and therefore cannot enter other parts of the central nervous system.
The composition of the present invention can be administered with a second pharmaceutical composition. By “both”, the composition can be formulated with other compounds within the same composition, and / or the other compound and / or pharmaceutical composition is prior to administration of the composition of the invention. Means administered during (including simultaneous administration) and / or after administration.

  Thus, in one embodiment, the composition of the present invention can be administered with a second pharmaceutical composition. The compositions can be administered as separate compositions or together in a single dosage form, simultaneously or sequentially as two different pharmaceutical compositions.

  In one embodiment, the pharmaceutical composition of the present invention comprises a selective serotonin reuptake inhibitor (SSRI), a serotonin noradrenaline reuptake inhibitor (SNRI), a norepinephrine serotonin reuptake inhibitor (NSRI), a selective noradrenaline reuptake inhibitor. Non-selective monoamine reuptake inhibitors, selective reversible monoamine reuptake inhibitors, including inhibitors, tetracyclic antidepressants, tricyclic antidepressants (TCAs), and other mechanisms of action It is administered with a second pharmaceutical composition selected from a pharmaceutical composition of an antidepressant, eg, an antidepressant such as mirtazapine. Specific examples of SSRIs include citalopram, escitalopram, fluoxetine, fluvoxamine, paroxetine and sertraline. A specific example of SNRI is venlafaxine. A specific example of NSRI is milnacipran.

Treatment method The treatment method is aimed at reducing the symptoms of gastrointestinal diseases.
The composition is preferably administered after feeling symptoms. For example, 5 minutes after symptom detection, for example 10 minutes after symptom detection, for example 15 minutes after symptom detection, for example, 20 minutes after symptom detection, for example, 30 minutes after symptom detection, for example 45 minutes from symptom detection By later, for example, 60 minutes after symptom sensing, for example, 75 minutes after symptom sensing, for example, 90 minutes after symptom sensing, for example, 120 minutes after symptom sensing, for example 150 minutes from symptom sensing It is administered later, for example, 180 minutes after symptom detection.
The composition can be administered once a day, every other day, twice a week, or once a week. The pharmaceutical composition is administered once a day, twice a day, or three times a day.

  The pharmaceutical composition is administered before a meal, for example 5 minutes before a meal, for example within 20 minutes of a meal, for example within 60 minutes of a meal.

  The pharmaceutical compositions disclosed herein can be prepared in unit dosage forms. In such form, the preparation is subdivided into unit dose forms containing appropriate quantities of the active component. A unit dosage form is a packaged preparation, a package containing discrete quantities of the preparation, eg, individually packaged powders. In this embodiment, the powder can be mixed with a solvent before or during use.

In one aspect of the invention, a suitable dose of the composition described herein is administered to an individual in need thereof for treatment in a pharmacologically effective amount. Here, the “pharmacologically effective amount” means administration of the total amount of each active ingredient of the pharmaceutical composition or the pharmaceutical composition or method in order to show a significant effect on the patient. As used herein, the term “unit dosage form” means a physically separated unit suitable as a single dose for human and animal subjects, each unit comprising a pharmaceutically acceptable diluent, carrier or medium. Together with a predetermined amount of the compound, alone or in combination with other drugs, calculated as an amount sufficient to obtain the desired medicinal effect. The details of the unit dosage form of the invention depend on the particular compound or compounds used, and the effects obtained and the pharmacodynamics associated with each compound in the host. A dose is an “effective amount” or the amount necessary to achieve an “effective level” for an individual patient.
The pharmaceutical composition can be administered by intravenous infusion. The infusion time is within 120 minutes, such as 100 minutes or less, such as 80 minutes or less, such as 60 minutes or less, such as 40 minutes or less, such as 20 minutes or less, such as 10 minutes or less, such as 5 minutes or less.

The administration requirements vary depending on the individual drug composition used, the method of administration and the subject to be treated. Ideally, patients treated with the methods of the invention are administered a pharmacologically effective amount of the compound that does not exceed the maximum tolerated dose (MTD), which is generally not greater than that required to develop drug resistance. .
Suitable dosage regimes are preferably factors well known in the art, such as the type, age, weight, sex, and condition of the subject being administered; route of administration; renal function and liver function of the subject; desired efficacy And the particular drug to be used.
Achieving drug concentrations within a range that exhibits drug efficacy without toxicity requires a regimen based on the kinetics of the drug's availability to the target site. This requires consideration of drug distribution, equilibrium and excretion. For the present invention, the dosage will vary depending on the drug used and the mode of administration.

  While the dose can be calculated based on the subject's body weight, under certain circumstances, the dose is calculated based on the subject's lean body mass (FFM). Thus, the dose is the concentration equivalent of PYY1-36 or PYY3-36.

In the present invention, the dosage will vary depending on the compound used and the mode of administration. The dose level varies between about 4 ng / kg body weight and 20 μg / kg body weight per day, preferably between about 10 ng / kg body weight and 1 μg / kg body weight, more preferably between 50 and 750 ng / kg body weight. To do. Another dose related to FFM is between about 5 ng / kg FFM to 25 μg / kg FFM, preferably between about 12.5 ng / kg FFM to 1.25 μg / kg FFM, more preferably 62.5 to 875 ng per day. / Kg FFM. To obtain a dose related to FFM, dose / kg body weight should be multiplied by a factor of 1.25.
The dosage can be administered as needed, for example up to 10 times a day, for example 1 to 5 times a day, for example 2 to 3 times a day, or preferably for example once a day. Therefore, the dose per day is 2 to 5 times the dose described above. Alternatively, the dosage can be administered less frequently than once a day as described above.

  A suitable dose of the composition used in the present invention is a concentration equivalent of PYY or a functional equivalent thereof corresponding to 4 ng to about 20 μg / kg body weight, or 10 ng to 1 μg / kg body weight, more preferably 50 ng to 750 ng / kg body weight. It is. The dose of PYY is preferably 20-200 ng / kg, 20-160 ng / kg, 40-160 ng / kg, 40-120 ng / kg, 40-80 ng / kg, 60-120 ng / kg, or about 60 ng / kg or 80 ng / kg. kg.

  Preferred doses are 1 pmol / kg to 5 nmol / kg, 5 pmol / kg to 1 nmol / kg, 20 pmol / kg to 500 pmol / kg, alternatively 40 to 160 pmol / kg, alternatively 75 to 120 pmol / kg of PYY or a functional equivalent thereof. Corresponding concentration. PYY1-36 can preferably be administered at a dosage of 50-400 pmol / kg, 80-320 pmol / kg, 150-250 pmol / kg, or about 200 pmol / kg. PYY3-36 can be administered preferably at a dosage of 30-350 pmol / kg, 50-280 pmol / kg, 80-200 pmol / kg, or about 120 pmol / kg. The dose for FFM can be calculated by multiplying the above dose by 1.25.

In a second aspect, the dosage is 5-50 pmol / kg, 5-40 pmol / kg, 5-30 pmol / kg, 10-40 pmol / kg, 10-30 pmol / kg, 5-25 pmol / kg, 5-20 pmol / kg. Yes, most preferably 10-20 pmol / kg, 15-30 pmol / kg, or about 15 pmol / kg or 20 pmol / kg.
The dosage is preferably administered once a day, twice a day, three times a day, four times a day, five times a day, or five times or more per day.
In one preferred aspect of the invention, the composition comprises from about 400 ng to about 2 mg, more preferably from about 10 μg to about 200 μg, alternatively from about 5 μg to about 250 μg, more preferably from about 20 μg to about 200 μg, more preferably from 20 μg to about 100 μg. At a dose of PYY or a functional equivalent thereof. Most preferably, the dose is 1-20 μg, 2-16 μg, 4-16 μg, 4-12 μg, 4-8 μg, 6-12 μg, 6-10 μg or about 8 μg.

  In preferred embodiments, the composition is administered at a dose of 100 pmol to 500 nmol, alternatively 500 pmol to 100 nmol, alternatively 1 nmol to 50 nmol, alternatively 2 to 25 nmol, alternatively 4-20 nmol, PYY or a functional equivalent thereof. Alternatively, preferred doses are 0.25-5 nmol, 0.5-4 nmol, 1-4 nmol, 1-3 nmol, 1-2 nmol, 1.5-3 nmol, or more preferably 1.5-2.5 nmol, Most preferably about 2 nmol. In a more preferred embodiment, the dose of PYY1-36 is, for example, 5-40 nmol, 8-32 nmol, 15-25 nmol, about 20 nmol, while PYY3-36 is 3-35 nmol, 5-28 nmol, 8-200 nmol, or about 12 nmol.

  In other embodiments, PYY or a functional equivalent thereof is 5-30 pmol / kg, 5-25 pmol / kg, 5-20 pmol / kg, or 10 to achieve effective levels in the individual being treated. It is administered subcutaneously at a dose of -20 pmol / kg. Currently preferred doses are 10-20 pmol / kg body weight for PYY1-36 or PYY3-36. In a second preferred embodiment, the dose of PYY1-36 is 150-250 pmol / kg and / or the dose of PYY3-36 is 80-150 pmol / kg.

The dose of PYY or a functional equivalent thereof is preferably administered once a day, twice a day, three times a day, four times a day, or five times a day.
The pharmaceutical compositions disclosed herein can be prepared as unit dosage formulations. In such dosage forms, the formulation is divided into single dosage forms containing a suitable amount of the active ingredient. Unit dosage forms are packaged preparations, packages containing separate quantities of the preparation, eg, individually packaged powders. In this embodiment, the powder can be mixed with the solvent before or during use.

  In a preferred embodiment, the PYY composition is administered in unit dosage form and is about 400 ng to about 2 mg, more preferably about 10 μg to about 200 μg, alternatively about 5 μg to about 250 μg, more preferably about 20 μg to about 200 μg, more preferably Administer in an amount of about 20 μg to about 100 μg of PYY or a functional equivalent thereof. Unit dosage forms contain 100 pmol to 500 nmol, alternatively 500 pmol to 100 nmol, alternatively 1 nmol to 50 nmol, alternatively 2-25, alternatively 4 nmol to 200 nmol PYY or a functional equivalent thereof. The composition is preferably administered once a day, twice a day, three times a day, four times a day, or five times a day. In a further preferred embodiment, the unit dose of PYY1-36 includes 5-40 nmol, 8-32 nmol, 15-25 nmol, about 20 nmol, and the unit dose of PYY3-36 is 3-35 nmol, 5-28 nmol, 8-200 nmol. About 12 nmol.

In certain embodiments, the pharmaceutical composition is administered by infusion. The dose of PYY or its functional equivalent for infusion is 0.01 pmol / kg min to 500 pmol / kg its functional equivalent, for example 0.05 pmol / kg min to 100 pmol / kg min, 0.1 pmol / kg min To 50 pmol / kg min, or 1 pmol / kg min to 25 pmol / kg min.
The composition is preferably administered after feeling symptoms, for example 5 minutes after symptom sensing, eg 10 minutes after symptom sensing, eg 15 minutes after symptom sensing, eg 20 minutes after symptom sensing, eg from symptom sensing Up to 30 minutes later, for example 45 minutes after symptom sensing, for example 60 minutes after symptom sensing, for example 75 minutes after symptom sensing, for example 90 minutes after symptom sensing, for example 120 minutes after symptom sensing, For example, administration is performed until 150 minutes after symptom detection, for example, 180 minutes after symptom detection.
The PYY composition of the present invention can be administered in admixture with a pharmacologically acceptable carrier or diluent.
Treatment includes doing it at night. Furthermore, since “effective levels” are used as the preferred endpoint of administration, the actual dose and administration schedule will vary with individual differences in pharmacokinetics, drug distribution, and metabolism. An “effective level” is defined as the desired blood or tissue level of a patient, for example, corresponding to the concentration of one or more compounds of the invention.
By effective level is meant the amount of active ingredient of the composition of the present invention that is capable of obtaining a certain blood or tissue level of the desired compound in the patient. An effective level is the amount of active ingredient of the composition of the present invention that can eliminate the patient's symptoms.
The pharmaceutical composition can be administered during hospitalization. Furthermore, if the composition can be self-administered, it is beneficial to the patient.
In one aspect of the invention, the composition of the invention can be self-administered. The pharmaceutical composition can be administered using an injection device, for example using a system similar to an insulin pen.
In one aspect of the invention, the composition can be administered using a single dose or multiple dose injection device.
The pharmaceutical composition comprises an antidepressant such as a selective serotonin reuptake inhibitor (SSRI), a serotonin / noradrenaline reuptake inhibitor (SNRI), a norepinephrine / serotonin reuptake inhibitor (NRSI), a selective noradrenaline reuptake. Anti-selective monoamine reuptake inhibitors, including inhibitors, tetracyclic antidepressants, tricyclic antidepressants (TCAs), selective reversible monoamine reuptake inhibitors, and other mechanisms of action such as mirtazapine It can also be a kit-in-part formulation further containing a depression. Specific examples of SSRIs include citalopram, escitalopram, fluoxetine, fluvoxamine, paroxetine and sertraline. A specific example of SNRI is venlafaxine. The kit-in-part formulation can be used simultaneously, sequentially or for separate administration. A specific example of NSRI is milnacipran.
In a preferred embodiment, the pharmaceutical composition is a kit-in-part formulation.

Examples The following examples illustrate the present invention, but are not limited thereto.

Combined and functional analysis
Transfection and tissue culture:
COS-7 cells can be cultured in Dulbecco's Modified Eagle Medium 1885 containing 10% fetal bovine serum, 2 mM glutamine and 0.01 mg / ml gentamicin. Expression plasmids containing cDNA encoding wild type or mutant receptors can be transiently expressed after transfection using the calcium phosphate precipitation method and analyzed 48 hours after transfection.

Binding analysis:
One day after transfection, cells are transferred to a multi-well plate for analysis and seeded. The number of cells seeded per well is selected to give 5-10% binding of the added radioligand. Two days after transfection, the cells are analyzed in a competitive binding test using ¹²⁵ I-PYY (3-36) as a tracer. Radioligand binds in a buffer consisting of 0.5 ml of 50 mM HEPES buffer (pH 7.4) supplemented with 1 mM CaCl ₂ , 5 mM MgCl ₂ , 0.1% BSA, and replaces unlabeled ligand in a dose-dependent manner. . The analysis was performed twice at 4 ° C. for 3 hours, and the analysis was stopped by washing twice with a buffer solution. Cell-added receptor-bound radioligand is measured by adding lysis buffer (48% urea, 3% acetic acid containing 2% NP-40). The concentration of radioligand in the analysis solution corresponds to a final concentration of about 20 pM.

Functional analysis:
COS-7 is cultured as described above and co-transfected. Activation of phospholipase C by a chimeric G-protein (Conklin B) formed between both Gαq and GαiY2 receptors can be measured through intracellular turnover of inositol phosphate (IP). IP turnover can be recorded using the following assay.
One day after transfection, COS-7 cells were cultured in medium supplemented with [ ³ H] myo-inositol (Amersham, PT6-271) 5: Ci and 10% fetal calf serum, 2 mM glutamine and 0.01 mg / mg gentamicin. Incubate for 24 hours in 1 ml per well. The cells were washed twice with 20 mM HEPES buffer (pH 7.4) containing 140 mM NaCl, 5 mM KCl, 1 mM MgSO ₄ , 1 mM CaCl ₂ , 10 mM glucose, 0.05% (w / v) bovine serum, and containing 10 mM LiCl. Incubate in 0.5 ml of buffer at 37 ° C. for 30 minutes. Incubate for a further 30 minutes with 1 U / ml concentration of adenosine deaminase ADA (200 U / mg, Boehringer Mannheim, Germany). After stimulation with various concentrations of peptides at 37 ° C. for 45 minutes, the cells are extracted with 10% ice-cold perchloric acid and incubated on ice for 30 minutes. The obtained supernatant is neutralized with a KEP solution in HEPES buffer, and the resulting [ ³ H] -inositol phosphate is purified with Bio-Rad AG 1-X18 anion exchange resin. The measurement is performed in duplicate.

Synthetic Preparation of PYY and Functional Equivalents The polypeptides of the present invention can be prepared by conventional peptide synthesis methods.
Amino acid derivatives and synthetic reagents are obtained from commercial sources. Peptide chain elongation is performed mainly using an Applied Biosystem 433A synthesizer (manufactured by Perkin Elimer). The protected peptide derivative resin is constructed by the Boc or Fmoc method. The protected peptide resin obtained by the Boc method is anhydrous hydrogen fluoride (HF) and deprotected in the presence of p-cresol to release the peptide, which is then purified. The protected peptide resin obtained by the Fmoc method is deprotected by dilution with trifluoroacetic acid (TFA) or various scavenger-containing TFAs, and the released peptide is purified. Purification is performed by C4 or C18 column reverse phase HPLC. The purity of the purified product is confirmed by reverse phase HPLC, and its structure is confirmed by amino acid composition analysis and mass spectrometry.

The PYY plasma level measurement experiment is performed by subcutaneous injection with placebo and PYY1-36 or PYY3-36 in four concentrations (shown in Table 1). The dose of PYY is calculated based on the subject's lean body mass (FFM).

結果は平均±ＳＥ、対応のあるｔ−検定（ＳＡＳ）および反復測定（ＳＡＳ）で示す。

Results are presented as mean ± SE, paired t-test (SAS) and repeated measures (SAS).

  PYY injection was performed at 0 minutes, and plasma concentrations of PYY after administration of PYY1-36 and PYY3-36 were t = 0, 15, 30, 45, 60, 75, 90, 120, 150, 180, 210, and 240. Measure in minutes.

PYY Analysis The plasma concentration of PYY is measured by PYY radioimmunoassay. Analysis is performed using PYY antiserum (code number: 8412-5) (Euro-Diagonistica, Malmoe, Sweden). The antiserum recognizes both human PYY1-36 and PYY3-36. Synthetic human PYY1-36 (Peninsula, Merseyside, UK) and porcine ¹²⁵ I-PYY (code number: IM259) are obtained from Amersham Bioscience, Buckinghamshire, UK and used as standards. The measurement limit of analysis is 2 pmol / l or less, and 50% inhibition is obtained at 40 pmol / l PYY. The recovery of PYY added to plasma at concentrations between 5 and 50 pmol / l is within 15% of the expected value. The deviation coefficient between experiments is 5% or less. The antiserum does not cross-react with human NPY or human PP at concentrations up to 500 pmol / l.
The results are shown in FIG. 2 and FIG.

In vivo measurement of the effect of PYY on migrating myoelectric complexes
Method:
Rats are equipped with bipolar electrodes 5 (duodenum), 15 and 25 (jejunum) cm distal from the pylorus for electromyography of the small intestine. The native ligand peptide YY1-36 and peptide YY3-36 are infused intravenously for 60 minutes at a dose of 0.5-400 pmol / kg min. The mechanism of action is examined in left and right vagus nerve isolated animals after pretreatment with 1 mg / kg of N-nitro-L-arginine (L-NNA) and 3 mg / kg of guanethidine.
The experimental protocol was approved by the local ethics committee for animal experiments in Northern Stockholm (Sweden).

material:
Male SD (Sprague-Dawley) rats (B & K, Sollentuna, Sweden) weighing 300-350 g were used in the experiments.

Preparation of rats for electromyography:
Rats are anesthetized with intraperitoneal administration of 50 mg / kg pentobarbital (Apoteksbolaget, Umea, Sweden). A midline incision with 3 bipolar stainless steel electrodes (SS-5T, Clark Electromedical Instruments, Reading, UK) and small intestinal muscles 5 (D), 15 (J1) and 25 (J2) cm distal to the pylorus Embed in the wall. All animals are equipped with a jugular vein catheter for drug administration. The electrodes and the catheter are guided outside the body through the subcutaneous part of the neck of the animal. After surgery, the animals are individually caged and allowed to recover for at least 7 days until the experiment is performed. During the recovery period, rats are trained to accept experimental conditions. The experiment is then carried out on animals that have been awakened after 8 hours of fasting in a wire-bottomed cage that allows free access to water. During the experiment, rats are placed in a Bollman cage. The electrodes are connected to an EEG preamplifier (7P5B) operating a Grass Polygraph 7B (Grass Instruments, Quincy, Mass.). The time constant is set to 0.015 seconds, and the low and high cutoff frequencies are set to 10 and 35 Hz, respectively.

Design of the electromyogram test All experiments begin by recording as a control the basal myoelectric activity of the four activity fronts that propagate over all three recording sites for 60 minutes.
Immediately after the fourth active front passes through the first electrode site, instillation is initiated using a microinjection pump (CMA 100, Carnegie Medicine, Stockholm, Sweden).
In the first series of experiments, a dose of 0.5 to 400 pmol / kg min of natural ligand PYY1-36 or PYY3-36 is administered intravenously for 60 minutes.
In a second series of experiments, 100 pmol / kg PYY1-36 or 45 minutes prior to infusion of PYY3-36 for min, NO synthase inhibitors of 1 mg / kg bolus injection N ^omega - nitro -L- arginine (L-NNA ).
In a third experimental series, 3 mg / kg guanethidine is administered on day 1 and the next day the effect of 100 pmol / kg min of PYY1-36 or PYY3-36 is examined.
In a fourth experimental series, the effect of infusion of 100 pmol / kg min of PYY1-36 or PYY3-36 is investigated in vagus and pseudovagus isolated animals.

Drug rats PYY1-36 and rat PYY3-36 are from Neosystem, Strasbourg, France, L-NNA is from Sigma-Aldrich (Sigma-Aldrich, St. Louis, MO, USA), and guanethidine is from Apoteksbolaget ( (Stockholm, Sweden).

Data analysis The main characteristics of fasting small intestine myoelectric activity, activity front, or phase III of MMC is as a period of clearly distinguishable strong spike activity with an amplitude at least twice that of the preceding baseline Identify and propagate to the opposite side of the entire recorded segment, followed by a rest period, followed by MMC Phase I.
The percentage of the recording period occupied by the Phase III activity front is calculated as the stimulation period (60 minutes). The effect of the peptide is expressed as a percentage of the time occupied by the activity front.
Dose response curves are obtained with Graph-Pad Prism 4 (GraphPad, San Diego, CA, USA). Data are expressed as mean ± SEM. Differences between each data group are measured by ANOVA, and if necessary, by Dunnett's test or Student's T test with statistical significance of p <0.05.

Clinical Protocol 45 subjects who meet the proposed diagnostic criteria for gastrointestinal disease IBS or functional dyspepsia will participate in this study.
Experiments will be performed in a double-blind, placebo-controlled trial format. Subjects are divided into three groups (each n = 15), Group A, Group B, and Group C. Subjects should write their symptoms (fecal habits / pain / nausea, etc.) and diet information in a diary. The first phase of the experiment is 4 weeks (Run-in Phase), after which the subject is treated with one of the following three regimes: Subjects will keep a diary of the timing, type and severity of symptoms experienced during the treatment phase. The duration of the treatment phase is 4 weeks).

Subjects in Medication A group receive placebo subcutaneous injections (Nacl) three times a day (equally distributed in waking hours). Subjects in group B received subcutaneous injections of 60 pmol / kg (body weight) PYY3-36 three times a day, and subjects in group C received subcutaneous injections of 100 pmol / kg (body weight) PYY3-36 three times a day Get an injection.
The subject's diary is checked by researchers, and the severity of symptoms is determined by food intake and timing.
The effect on symptoms was compared with the results of the treatment phase for 4 weeks and the results of the introduction phase for each subject, and then statistically compared the results of Group B with Group A and the results of Group C with Group A. To evaluate.

Effect of PYY on pain sensation and rectal locomotor activity Twelve patients (men and women between the ages of 18 and 60) who meet the Roma II diagnostic criteria will participate in this study.
Operation of the experiment All patients come to the clinic after 12 hours of fasting. Prior to the experiment, the patient undergoes a tap water enema.
A probe (barostat bag) is inserted into the rectum and automatically connected to an electric pressure regulator (barostad) programmed to inflate at regular intervals and with a constant increase in bag pressure.
The experiment is performed as three crossover studies of approximately 7 days between treatments.
A) Subcutaneous injection of 0.9% NaCl solution of PYY1-36 B) Subcutaneous injection of 0.9% NaCl solution of PYY3-36 C) Subcutaneous injection of saline (0.9% NaCl solution)

Swelling begins following a defined protocol after administration of PYY or saline. Patients record discomfort and pain using a visual analog scale (VAS). The patient is allowed to be able to deflate the bag immediately in case of severe discomfort or pain, thereby allowing the protocol to be controlled. Write down pressure and volume thresholds.
After a period of time, for example after 20 or 30 minutes, a saline meal with a constant energy content is given by mouth.
Thereafter, a second expansion similar to the first expansion starts.
A constant low pressure is maintained in the pressure regulating bag (barostat bag) during the two inflations. This makes it possible to record bag volume as a measure of rectal motor activity.

Comparison of pain threshold with or without food intake in patients with primary endpoint IBS and comparison of PYY effect with placebo
Second endpoint: A comparative study of the physiological effects of PYY on locomotor activity in the rectum with placebo

The effect on symptoms is assessed by statistically comparing the results of Group B with Group A and the results of Group C with Group A.
Statistical analysis is done by nonparametric analysis of differences (Kruskal-Wallis test).

Structure of PP-fold peptide family Plasma concentration of PYY1-36 Plasma concentration of PYY3-36

Claims (21)

  A composition containing PYY or a functional equivalent thereof, or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating functional gastrointestinal diseases. The functional gastrointestinal disorder is irritable bowel syndrome,
a) the main stool habit is constipation, and / or b) diarrhea and constipation in which the main stool habit alternates, and / or c) the treatment is administered parenterally,
The composition according to claim 1.   The composition of claim 1 for the treatment of abdominal pain.   4. A composition according to claim 3 for the treatment of visceral pain.   The composition according to claim 3 or 4, wherein the functional gastrointestinal disease is irritable bowel syndrome or functional dyspepsia.   The composition of claim 1, wherein the functional gastrointestinal disorder is functional dyspepsia. 7. A composition according to claim 6 for the treatment of the following symptoms:
a) satiety, and / or b) unable to eat a standard amount of meal, and / or c) pain after food intake, and / or d) nausea and / or e) vomiting, and / or f A) abdominal distension, and / or g) stagnation, and / or h) reflux, and / or i) upper abdominal pain, and / or j) bloating, and / or k) excess intestinal gas,
And any combination of the above symptoms.   A composition according to any preceding claim, wherein the symptoms are mainly upper gastrointestinal symptoms.   A composition according to any preceding claim, wherein the symptoms are mainly lower gastrointestinal symptoms.   A composition according to any preceding claim, comprising human PYY.   The composition according to any of the preceding claims, comprising human PYY1-36.   40. A composition according to any preceding claim, comprising human PYY3-36.   A composition according to any preceding claim, wherein the pH of the composition is between 2.0 and 9.0.   A composition according to any preceding claim, wherein the pharmaceutical composition is for parenteral administration.   A composition according to any preceding claim, wherein the pharmaceutical composition is for subcutaneous administration.   A composition according to any preceding claim, wherein the pharmaceutical composition is for intranasal administration.   A composition according to any preceding claim, wherein the composition contains a second active ingredient.   The second pharmaceutical composition comprises SSRI (citalopram, escitalopram, fluoxetine, fluvoxamine, paroxetine and sertraline), SNRI (venlafaxine), NSRI, tetracyclic antidepressant, tricyclic antidepressant (TCA) 19. The composition of claim 18, which is a non-selective monoamine reuptake inhibitor, a selective reversible monoamine reuptake inhibitor and an antidepressant such as mirtazapine.   19. A composition according to claim 18, wherein the second pharmaceutical composition is an antiemetic.   The composition according to claim 20, which is used for treatment of IBS and / or FD.   A method of treatment, comprising administering a pharmaceutical composition according to any of the preceding claims.

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