Amylin analogues

Abstract

A compound or derivative of a compound comprising a peptide chain that is an analog of amylin, a terminal -NH2 group attached to the C-terminus of the peptide chain, and an alkylene or alkenylene chain attached to the other end of the peptide chain; or a salt or solvate of the compound or derivative. The compounds, derivatives, and salts and related compositions are suitable for preventing or treating diabetes, obesity, heart disease, stroke, and non-alcoholic fatty liver disease, improving insulin release in a subject, improving carbohydrate metabolism in a subject, improving the lipid profile of a subject, improving carbohydrate tolerance in a subject, reducing appetite, reducing food intake, and / or reducing calorie intake, and related purposes.

Description

[Technical Field]

[0001] The present disclosure relates to compounds that are peptide hormone analogs and are useful in the treatment of disorders such as diabetes and obesity. [Background technology]

[0002] According to the National Health and Nutrition Examination Survey (NHANES, 2009-2010), 33.0% of U.S. adults aged 20 years and older were overweight, 35.7% were obese, and 6.3% were severely obese. Since then, the prevalence of obesity in the United States has further increased, and in 2020, 41.9% of U.S. adults were classified as obese and 9.2% were classified as severely obese (NHANES, 2020). Furthermore, a significant number of U.S. children are overweight or obese.

[0003] The causes of obesity are complex and multifactorial. Increasing evidence suggests that obesity is not a simple matter of self-control, but a complex disorder involving appetite suppression and energy metabolism. Furthermore, obesity is associated with a variety of conditions that are associated with increased morbidity and mortality in the population. The etiology of obesity has not been clearly established, but genetic, metabolic, biochemical, cultural, and psychosocial factors are thought to contribute. Generally, obesity is described as a condition in which excess body fat places an individual at risk for health.

[0004] There is strong evidence that obesity is associated with increased morbidity and mortality. Disease risk, such as cardiovascular disease risk and type 2 diabetes risk, increases independently with increasing body mass index (BMI). Indeed, this risk has been quantified as a 5 percent increase in heart disease risk for women and a 7 percent increase in heart disease risk for men for each point of BMI above 24.9 (see Kenchaiah et al., N. Engl. J. Med. 347:305, 2002; Massie, N. Engl. J. Med, 347:358, 2002).

[0005] Diabetes mellitus (DDM) is a chronic syndrome in which carbohydrate, protein, and fat metabolism is impaired due to insufficient insulin secretion or insulin resistance in target tissues. It occurs in two major forms: insulin-dependent diabetes mellitus (type 1 diabetes) and non-insulin-dependent diabetes mellitus (type 2 diabetes). Type 1 diabetes, or insulin-dependent diabetes mellitus (IDDM), is caused by beta cell destruction, leading to insufficient endogenous insulin levels. Type 2 diabetes, or non-insulin-dependent diabetes mellitus, results from both a lack of the body's sensitivity to insulin and a relative deficiency in insulin production. According to the 2014 National Diabetes Statistics Report, approximately 28.9 million adults aged 20 years and older in the United States have diabetes (based on estimates from the 2009–2012 National Health and Nutrition Examination Survey applied to 2012 U.S. Census data). Among adults, 90–95% of diabetes cases are type 2 diabetes.

[0006] There is considerable evidence that weight loss in obese individuals reduces important disease risk factors. For example, even small weight losses, such as 10% of initial body weight, in both overweight and obese adults have been associated with reduced risk factors such as hypertension, hyperlipidemia, and hyperglycemia. Significant weight loss has been shown to effectively reverse type 2 diabetes (Lim et al., Diabetologia June 2011).

[0007] Although diet and exercise provide simple methods for reducing weight gain, overweight and obese individuals often cannot adequately control these factors to effectively reduce weight. Pharmacological treatments are available; several weight-loss drugs have been approved by the Food and Drug Administration and can be used as part of a comprehensive weight-loss program. However, many of these drugs have serious adverse side effects. When less invasive methods fail and patients are at high risk of obesity-related morbidity or mortality, weight-loss surgery is an option for carefully selected patients with severe clinical obesity. However, these procedures are high-risk and suitable for use in only a limited number of patients. Obese subjects are not the only ones who want to lose weight. People who weigh within the recommended range, for example, at the upper end of the recommended range, may want to reduce their weight and approach their ideal weight. Therefore, there remains a need for agents that can be used to cause weight loss in overweight and obese subjects and normal-weight subjects.

[0008] Several approaches to the development of drugs useful in inducing weight loss involve gastrointestinal peptide hormones and their analogs.

[0009] However, despite considerable progress, identifying substances useful as drugs remains complex and often unpredictable. To be useful as a therapeutic agent, a compound must have an appropriate range of properties. In addition to having good efficacy at the intended biological target, a compound must have good in vivo pharmacokinetic properties, low toxicity, and an acceptable side effect profile. For example, even with commercially available drugs such as liraglutide, side effects may include nausea and vomiting, and concerns have been raised regarding thyroid cancer and pancreatitis.

[0010] Thus, there remains a need for additional compounds useful for treating disorders and diseases such as diabetes and obesity. For example, it would be desirable to identify peptides that have beneficial properties, such as improved activity profiles, and / or reduced side effects. For example, it would be desirable to identify peptides that increase energy expenditure in a subject but do not significantly reduce food intake. If a compound does not significantly reduce food intake, the compound is expected to have fewer side effects, such as nausea. Alternatively, or additionally, it would be desirable to identify peptides that have these and other biological effects over an extended period of time. Compounds with a longer duration of activity can be administered less frequently and in lower doses, which contributes to improved convenience for the subject, fewer side effects, and lower costs.

[0011] Amylin (also known as islet amyloid polypeptide, IAPP) is a 37-residue peptide hormone co-secreted with insulin by pancreatic beta cells. It is known to play a role in blood glucose regulation, slow gastric emptying, and increase satiety. Its production may be deficient in subjects with type 2 diabetes who have lost pancreatic beta cells.

[0012] Human amylin has amyloidogenic properties, meaning that it has the ability to form insoluble precipitates and fibrils both intracellularly and extracellularly. These amyloid fibrils and precipitates are known to be cytotoxic and have been implicated in both neurodegeneration and pancreatic beta cell death.

[0013] Amyloidogenic activity is present in human amylin and, at least to some extent, in amylin from other primates. Amylin from other species, including mouse and rat, lacks residues responsible for amyloidogenic activity. Amylin Pharmaceuticals, Inc. markets the drug pramlintide acetate (Symlin). (商標)), which is an acetate salt of an analog of human amylin in which residues 25, 26, and 29 of human amylin are substituted to reduce (though not completely eliminate) its amyloidogenic activity. Pramlintide acetate was approved by the FDA in 2005 for use in diabetic patients who use insulin but have difficulty maintaining glycemic control. This allows patients to use less insulin, resulting in reduced blood sugar undershoot and overshoot. It has satiety-promoting properties but is less effective at reducing unhealthy weight and is not approved for use as a weight-loss drug.

[0014] Novo Nordisk has a potential obesity management drug in clinical trials called caglilintide, which is co-administered with semaglutide. Caglilintide has a peptide sequence that is highly similar to that of pramlintide. Therefore, the sequences of all the peptides discussed are shown below: Pramlintide KCNTATCATQRLANFLVHSSNNFGPILPPTNVGSNTY-(NH2) (SEQ ID NO: 1) Caglilintide KCNTATCATQRLAEFLRHSSNNFGPILPPTNVGSNTP-(NH2) (SEQ ID NO: 2) Human amylin KCNTATCATQRLANFLVHSSNNFGAILSSTNVGSNTY-(NH2) (SEQ ID NO: 3) Rat amylin KCNTATCATQRLANFLVRSSNNLGPVLPPTNVGSNTY-(NH2) (SEQ ID NO: 4)

[0015] Amylin, caglilintide, and pramlintide are all amidated at their C-terminus. All of these peptides also have an -SS- bridge between the two C residues. For an injectable weight loss drug to be tolerated by patients and affordable, it preferably has a half-life after injection of at least several days, preferably one week or more, so that it can be conveniently administered at intervals of one week or more. All of the peptides listed above are short-lived. The drug caglilintide (not amylin or pramlintide) is lipidated to extend its half-life to the extent that a once-weekly injection regimen may be feasible.

[0016] There is still a need for improved therapeutic weight loss agents. Despite considerable progress, identifying substances useful as drugs remains complex and often unpredictable. To be useful as a therapeutic agent, a compound must have an appropriate range of properties. In addition to having good efficacy at the intended biological target, a compound must also have good in vivo pharmacokinetic properties, low toxicity, and an acceptable side effect profile. For example, even with commercially available drugs such as liraglutide, side effects may include nausea and vomiting, and there have been concerns about thyroid cancer and pancreatitis.

[0017] Thus, there remains a need for additional compounds useful for treating disorders and diseases such as diabetes and obesity. For example, it would be desirable to identify peptides with beneficial properties, such as improved activity profiles, and / or reduced side effects. For example, it would be desirable to identify peptides that increase energy expenditure in a subject but do not significantly reduce food intake. If a compound does not significantly reduce food intake, the compound is expected to have fewer side effects, such as nausea. Alternatively, or in addition, it would be desirable to identify peptides that have these and other biological effects over a longer period of time. Compounds with a longer period of activity can be administered less frequently and in lower doses, which contributes to improved convenience for the subject, fewer side effects, and lower cost. The present invention provides novel weight loss agents based on amylin but with advantages over caglilintide, including greater efficacy, a longer half-life, and lower cost. [Prior art documents] [Patent documents]

[0018] [Patent Document 1] U.S. Patent No. 4,179,337 [Patent Document 2] U.S. Patent No. 6,447,743 [Patent Document 3] U.S. Patent No. 5,700,486 [Patent Document 4] U.S. Patent No. 6,436,091 [Patent Document 5] U.S. Patent No. 5,939,380 [Patent Document 6] U.S. Patent No. 5,993,414 [Non-patent literature]

[0019] [Non-Patent Document 1] Kenchaiah et al., N. Engl. J. Med. 347:305, 2002 [Non-patent document 2] Massie, N.Engl.J.Med, 347:358, 2002 [Non-licensed document 3] National Diabetes Statistics Report, 2014

Non-licensed Document 4

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Non-licensed Document 6

Non-licensed Document 7

Non-licensed literature 9

Non-licensed literature 10

Non-licensed Document 11

Non-licensed Document 12

[0020] In a first aspect of the present invention, a compound of the formula: ABC or a salt or solvate of the compound or derivative, wherein: A is Q, Q-Ser, Q-Glu or Q-Lys, and Q is a group of formulas: [ka] and In the formula, R is C8 to C 28 an alkylene or alkenylene chain, R1 is -CO2H; B is a peptide moiety of 32 or 33 amino acid residues having the sequence XYZ, wherein: X is Lys-Cys, Y is Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6, wherein: Xaa1 is Ser or absent, Xaa2 is Asn, Lys, Ser, or Thr; Xaa3 is Thr or Leu; Xaa4 is Ala or Ser, Xaa5 is Thr, Xaa6 is Cys, Z is Xaa7-Xaa8-Xaa9-Xaa10-Xaa11-Xaa12-Xaa13-Xaa14-Xaa15-Xaa16-Xaa17-Xaa18-Xaa19-Xaa2 0-Xaa21-Xaa22-Xaa23-Xaa24-Xaa25-Xaa26-Xaa27-Xaa28-Xaa29-Xaa30-Xaa31 (SEQ ID NO: 6), in the formula: Xaa7 is Ala, Val or Met; Xaa8 is Thr or Leu; Xaa9 is Gln or Gly; Xaa10 is Arg or Lys; Xaa11 is Leu, Xaa12 is Ala or Ser, Xaa13 is any amino acid, Xaa14 is Glu, Asp or Phe; Xaa15 is Leu, Xaa16 is His, Xaa17 is Lys or Arg; Xaa18 is Leu, Xaa19 is Gln or Lys; Xaa20 is Thr, Xaa21 is Tyr or Phe; Xaa22 is Pro, Xaa23 is Arg or Lys; Xaa24 is Thr, Xaa25 is Gln, Asp, Pro, Lys, or Asn; Xaa26 is Val or Thr; Xaa27 is Gly, Xaa28 is Ser or Ala, Xaa29 is Lys, Asn, Gly or Asp; Xaa30 is Thr or Ala; Xaa31 is any amino acid, C is a terminal -NH2 group attached to the C-terminus of peptide moiety B.

[0021] Also provided herein are compositions comprising a compound, derivative, salt, or solvate of the invention together with a pharmaceutically acceptable carrier, and optionally, an additional therapeutic agent.

[0022] Also provided herein is a compound, derivative, salt, or solvate of the invention, or a composition comprising such a compound, derivative, salt, or solvate and a pharmaceutically acceptable carrier, for use as a pharmaceutical, for example, for the prevention or treatment of diabetes, obesity, heart disease, stroke, or non-alcoholic fatty liver disease, improving insulin release in a subject, improving carbohydrate metabolism in a subject, improving the lipid profile in a subject, reducing appetite, reducing food intake, reducing calorie intake, and / or improving carbohydrate tolerance in a subject.

[0023] Also provided herein are methods for treating or preventing a disease or disorder or other unwanted physiological condition in a subject, e.g., treating or preventing diabetes, obesity, heart disease, stroke, or non-alcoholic fatty liver disease in a subject, improving insulin release in a subject, improving carbohydrate metabolism in a subject, improving the lipid profile in a subject, improving carbohydrate tolerance in a subject, reducing appetite, reducing food intake, and / or reducing calorie intake in a subject, comprising administering a therapeutically effective amount of a compound, derivative, salt, or solvate of the invention, or a composition comprising such a compound, derivative, salt, or solvate and a pharmaceutically acceptable carrier.

[0024] Also provided herein is the use of a compound, derivative, salt, or solvate of the invention for the manufacture of a medicament for the prevention or treatment of diabetes, obesity, heart disease, stroke, or non-alcoholic fatty liver disease, improving insulin release in a subject, improving carbohydrate metabolism in a subject, improving the lipid profile in a subject, improving carbohydrate tolerance in a subject, reducing appetite, reducing food intake, and / or reducing calorie intake.

[0025] Also provided herein are methods for causing weight loss or preventing weight gain in a subject for cosmetic purposes, comprising administering an effective amount of a compound, derivative, salt, or solvate of the invention. [Brief explanation of the drawings]

[0026] [Figure 1]

[0023] Figure 1 shows, in a multi-page table, the amino acid sequences of exemplary compounds of the invention. The compounds are presented with the N-terminal residue on the left-hand side of the table. Amino acid sequences shown in Figure 1 that are not part of the compounds of the invention, such as prior art sequences and naturally occurring sequences, are included for ease of comparison only. DETAILED DESCRIPTION OF THE INVENTION

[0027] The results of the uptake experiment and the receptor activity experiment are shown in the right column of the table in FIG.

[0028] array Amino acid sequences herein are presented with the N-terminus at the left, and when a sequence spans multiple lines, the N-terminus is at the top left. Unless otherwise indicated, amino acid residues in a sequence are L-amino acids.

[0029] The amino acid sequences listed in this application are shown using standard letter abbreviations for amino acids.

[0030] The specific sequences shown herein relate to particular embodiments of the invention.

[0031] Detailed Description definition In order to facilitate review of the various embodiments of the disclosure, the following explanations of specific terms are provided:

[0032] Amylin: A 37-peptide hormone. In certain aspects of the present disclosure, the amylin can be mammalian amylin, such as rodent or human amylin.

[0033] Animal: Living multi-cellular vertebrate organisms, a category that includes mammals and birds. The term mammal includes both human and non-human mammals. Similarly, the term "subject" includes both human and veterinary subjects. In preferred embodiments of the invention, the subject is a human subject.

[0034] Appetite: The natural desire or craving for food. In one embodiment, appetite is measured by a survey to assess desire for food. Increased appetite generally leads to increased eating behavior.

[0035] Appetite suppressant: A compound that reduces the desire for food. Over-the-counter appetite suppressants include, but are not limited to, amfepramone (diethylpropion), phentermine, mazindol and phenylpropanolamine, fenfluramine, dexfenfluramine, and fluoxetine.

[0036] Body Mass Index (BMI): Sometimes called the Quetelet index, this is a mathematical formula for measuring body mass. BMI is calculated by multiplying weight (kg) by height. 2 (meters 2 ) The current standard accepted as "normal" for both men and women is 20-24.9 kg / m 2 In one embodiment, the BMI is 25 kg / m 2 A BMI above 25.5 kg / m can be used to identify obese subjects. Grade 1 obesity (which is sometimes referred to as being "overweight" rather than obese) is 25-29.9 kg / m 2 Grade II obesity corresponds to a BMI of 30-40 kg / m 2 Grade III obesity corresponds to a BMI of 40 kg / m2 (Jequier, Am. J Clin. Nutr. 45:1035-47, 1987). Ideal body weight varies among species and individuals based on height, build, bone structure, and sex.

[0037] Diabetes: The inability of cells to transport endogenous glucose across membranes due to either endogenous insulin deficiency and / or defective insulin sensitivity. Diabetes is a chronic syndrome in which carbohydrate, protein, and fat metabolism is impaired due to insufficient insulin secretion or insulin resistance in target tissues. It occurs in two main forms: insulin-dependent diabetes mellitus (IDDM, type I) and non-insulin-dependent diabetes mellitus (NIDDM, type II), which differ in etiology, pathogenesis, genetics, age of onset, and treatment.

[0038] The two major forms of diabetes are both characterized by the inability to deliver insulin in the amounts and at the precise times needed to control glucose homeostasis. Type 1 diabetes, or insulin-dependent diabetes mellitus (IDDM), is caused by the destruction of beta cells, resulting in insufficient levels of endogenous insulin. Type 1I diabetes, or non-insulin-dependent diabetes, results from a defect in both the body's sensitivity to insulin and a relative deficiency in insulin production. Preferred embodiments of the present invention may relate to the treatment of diabetes, including the treatment of type 1 diabetes or type 1I diabetes. In other embodiments, the present invention may relate to providing weight loss (including treating or preventing obesity or overweight conditions) in subjects with diabetes (type 1 or type 2) or prediabetes or other conditions or disorders characterized by poor glycemic control.

[0039] Energy Metabolism: The body must consume a certain amount of energy to maintain normal metabolism. In civilized humans, this is often considered to be approximately 2,800 calories per day. If food consumption does not provide this, weight loss results. However, energy metabolism is also regulated; for example, administration of glucagon is thought to increase metabolic rate, resulting in greater food intake being required to achieve energy balance and maintain body weight. Thus, if food intake is maintained at a normal level but energy metabolism is increased, weight loss results.

[0040] Food intake: The amount of food consumed by an individual. Food intake can be measured by volume or mass. For example, food intake can be the total amount of food consumed by an individual. In animal intake experiments, "food intake" is the mass of standardized chow consumed by the animal within a 24-hour period. Similar experiments can be conducted with human subjects by providing them with an unlimited buffet of calorie-controlled food portions or items and recording how much is consumed (e.g., by asking the human subjects to keep a record, by using third-party observation, or by placing each food portion or item in a separate dish and counting the number of empty dishes at the end of the experiment). Alternatively, food intake can be the amount of protein, fat, carbohydrates, cholesterol, vitamins, minerals, or any other dietary component of an individual. "Protein intake" refers to the amount of protein consumed by an individual. Similarly, "fat intake," "carbohydrate intake," "cholesterol intake," "vitamin intake," and "mineral intake" refer to the amount of protein, fat, carbohydrate, cholesterol, vitamin, or mineral consumed by an individual.

[0041] Normal daily diet: The average food intake of an individual of a given species. A normal daily diet can be expressed in terms of calorie intake, protein intake, carbohydrate intake, and / or fat intake. A normal daily diet for a human generally includes: from about 2,000, about 2,400, or about 2,800 to significantly more calories. Furthermore, a normal daily diet for a human generally includes about 12 g to about 45 g of protein, about 120 g to about 610 g of carbohydrates, and about 11 g to about 90 g of fat. A low-calorie diet is considered to be no more than about 85%, preferably no more than about 70%, of the normal calorie intake of a human individual.

[0042] In animals, calorie and nutrient requirements vary depending on the species and size of the animal. For example, in cats, the total calorie intake per pound and the percentage distribution of protein, carbohydrates, and fat vary depending on the age and reproductive status of the cat. However, a general guideline for cats is 40 calories / lb / day (18.2 calories / kg / day). About 30% to about 40% should be protein, about 7% to about 10% should be from carbohydrates, and about 50% to about 62.5% should come from fat intake. A typical daily diet for an individual of any species can be easily determined by one skilled in the art.

[0043] Obesity: A condition in which excess body fat can put people at health risks (see Barlow and Dietz, Pediatrics 102:E29, 1998; National Institutes of Health, National Heart, Lung, and Blood Institute (NHLBI), Obes. Res. 6(Suppl 2):51S-209S, 1998). Excess body fat is the result of an imbalance between energy intake and energy expenditure. For example, to assess obesity, the body mass index (BMI) can be used. One commonly used convention is a BMI of 25.0 kg / m 2 ~29.9kg / m 2 is overweight, but has a BMI of 30 kg / m 2 A BMI above this is considered obese.

[0044] Another convention uses waist circumference to assess obesity. In this convention, a waist circumference of 102 cm or greater is considered obese in men, while a waist circumference of 89 cm or greater is considered obese in women. Strong evidence indicates that obesity affects both an individual's morbidity and mortality. For example, obese individuals are at increased risk for, among other things, heart disease, non-insulin-dependent (type 2) diabetes, hypertension, stroke, cancer (e.g., endometrial, breast, prostate, and colon cancer), dyslipidemia, gallbladder disease, sleep apnea, reduced fertility, and osteoarthritis (see Lyznicki et al., Am. Fam. Phys. 63:2185, 2001).

[0045] Overweight: An individual whose weight exceeds their ideal body weight. An overweight individual may be obese, but is not necessarily obese. For example, an overweight individual is any individual who wishes to lose weight. In one convention, an overweight individual is someone with a BMI of 25.0 kg / m 2 ~29.9kg / m 2 This is an individual.

[0046] Peripheral administration: Administration outside the central nervous system. Peripheral administration does not include direct administration to the brain. Peripheral administration includes, but is not limited to, intravascular, intramuscular, subcutaneous, inhalation, oral, rectal, transdermal, or intranasal administration.

[0047] Polypeptide: A polymer of amino acid residues in which the monomers are linked via amide bonds. Polypeptides are sometimes referred to as "peptides." Unless otherwise indicated by context, the terms "polypeptide," "peptide," or "protein" as used herein encompass any amino acid sequence, including modified sequences such as glycoproteins. The terms "polypeptide" and "peptide" cover naturally occurring proteins as well as recombinantly or synthetically produced ones. The term "polypeptide fragment" refers to a portion of a polypeptide, e.g., a fragment that exhibits at least one sequence useful for receptor binding. The term "functional fragment of a polypeptide" refers to all fragments of a polypeptide that retain the activity of the polypeptide. Biologically functional peptides can also include fusion proteins in which the peptide of interest is fused to another peptide that does not diminish its desired activity. Generally, proteins are longer than peptides / polypeptides. The term "protein" is generally designated in reference to larger molecules, either a single chain of 100 or more amino acids or several chains of amino acids linked together.

[0048] Subcutaneous administration: Subcutaneous administration is the administration of a substance into the subcutaneous fat layer found between the dermis of the skin and the underlying tissue. Subcutaneous administration may be by injection, for example, using a hypodermic needle attached to a syringe or "pen" injection device. Other administration methods, such as microneedles, may also be used. Hypodermic needle injections typically involve some degree of pain for the recipient's benefit. Such pain can be mitigated by the use of local anesthetics or analgesics. However, typical methods used to reduce the perceived pain of injections are simply to distract the subject immediately before and during the injection. Pain can be minimized by using a relatively small gauge hypodermic needle, by injecting small amounts of substance, and by avoiding overly acidic or alkaline compositions, which may cause the subject to experience a "stinging" sensation at the injection site. Compositions having a pH between pH 4 and pH 10 are generally considered to be tolerably painless.

[0049] Therapeutically effective amount: A dose sufficient to prevent progression or cause regression of a disorder, or capable of alleviating a sign or symptom of a disorder, or otherwise achieving a desired result. In some embodiments, a therapeutically effective amount of a compound of the invention is an amount sufficient to inhibit or stop weight gain, or an amount sufficient to reduce appetite.

[0050] Compounds of the Invention The present inventors have found that the exemplified compounds of the present invention have properties such as causing weight loss in vivo. The compounds also have long serum half-lives, which means that the compounds can be administered conveniently infrequently.

[0051] Compared to naturally occurring human amylin and previous derivatives of amylin, including pramlintide and caglilintide, the compounds of the present invention have additional amino acid substitutions at one or more positions in the peptide sequence. Furthermore, the compounds of the present invention are lipidated with an aliphatic dicarboxylic acid. In many embodiments of the present invention, the aliphatic dicarboxylic acid is eicosane diacid. An aliphatic dicarboxylic acid (e.g., eicosane diacid) can be added to the N-terminus of a compound's peptide sequence via a condensation reaction between one of the acid groups of the aliphatic dicarboxylic acid and the alpha-amino group at the N-terminus of the compound's peptide sequence. Alternatively, an aliphatic dicarboxylic acid (e.g., eicosane diacid) can be added to an additional glutamic acid moiety via a condensation reaction between one of the acid groups of the aliphatic dicarboxylic acid and the amino group of glutamic acid, thereby conjugating glutamic acid to the N-terminal amino group of a compound's peptide sequence via its side chain carboxylic acid group (i.e., the group attached to the gamma carbon of glutamic acid).

[0052] The compounds of the present invention contain sequence substitutions and functionalizations, both alone and in novel combinations not previously investigated, and the beneficial properties discovered by the inventors have not been seen before. In some embodiments, these opportunities result in compounds that are up to three times more potent than caglilintide and have half-lives up to twice as long. Furthermore, the compounds of the present invention contain peptide moieties that are slightly shorter than those present in caglilintide. The use of shorter peptides reduces production costs and facilitates purification.

[0053] Peptide sequence As noted above, the compounds of the present invention have the formula: ABC It has.

[0054] A is Q, Q-Ser, Q-Glu or Q-Lys, and Q is a group of formulas: [ka] and In the formula, R is C8 to C 28 an alkylene or alkenylene chain, R1 is -CO2H; B is a peptide moiety of 32 or 33 amino acid residues having the sequence XYZ, wherein: X is Lys-Cys, Y is Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6, wherein: Xaa1 is Ser or absent, Xaa2 is Asn, Lys, Ser, or Thr; Xaa3 is Thr or Leu; Xaa4 is Ala or Ser, Xaa5 is Thr, Xaa6 is Cys, Z is Xaa7-Xaa8-Xaa9-Xaa10-Xaa11-Xaa12-Xaa13-Xaa14-Xaa15-Xaa16-Xaa17-Xaa18-Xaa19-Xaa2 0-Xaa21-Xaa22-Xaa23-Xaa24-Xaa25-Xaa26-Xaa27-Xaa28-Xaa29-Xaa30-Xaa31 (SEQ ID NO: 6), in the formula: Xaa7 is Ala, Val or Met; Xaa8 is Thr or Leu; Xaa9 is Gln or Gly; Xaa10 is Arg or Lys; Xaa11 is Leu, Xaa12 is Ala or Ser, Xaa13 is any amino acid, Xaa14 is Glu, Asp or Phe; Xaa15 is Leu, Xaa16 is His, Xaa17 is Lys or Arg; Xaa18 is Leu, Xaa19 is Gln or Lys; Xaa20 is Thr, Xaa21 is Tyr or Phe; Xaa22 is Pro, Xaa23 is Arg or Lys; Xaa24 is Thr, Xaa25 is Gln, Asp, Pro, Lys, or Asn; Xaa26 is Val or Thr; Xaa27 is Gly, Xaa28 is Ser or Ala, Xaa29 is Lys, Asn, Gly or Asp; Xaa30 is Thr or Ala; Xaa31 is any amino acid, C is a terminal -NH2 group attached to the C-terminus of peptide moiety B.

[0055] As can be seen, a significant portion of the compound consists of peptide sequence B, which can be 32 or 33 amino acid residues in length. It is preferably 32 amino acids in length.

[0056] Next, considering each of the residues in B, B has the sequence XYZ, where X is Lys-Cys; Y is Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6.

[0057] Xaa1 is Ser or absent. In a preferred embodiment, Xaa1 is absent. As a result, peptide sequence B is 32 amino acid residues long.

[0058] Xaa2 is Asn, Lys, Ser or Thr. Preferably, Xaa2 is Asn, Lys or Ser.

[0059] Xaa3 is Thr or Leu. Preferably, Xaa3 is Thr when Xaa1 is absent. Preferably, Xaa3 is Leu when Xaa1 is Ser.

[0060] Xaa4 is Ala or Ser. Preferably, Xaa4 is Ala when Xaa1 is absent. Preferably, Xaa2 is Ser when Xaa1 is Ser.

[0061] Xaa5 is Thr,

[0062] Xaa6 is Cys.

[0063] Preferably, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr and Xaa6 is Cys.

[0064] Z is Xaa7-Xaa8-Xaa9-Xaa10-Xaa11-Xaa12-Xaa13-Xaa14-Xaa15-Xaa16-Xaa17-Xaa18-Xaa19-Xaa2 0-Xaa21-Xaa22-Xaa23-Xaa24-Xaa25-Xaa26-Xaa27-Xaa28-Xaa29-Xaa30-Xaa31 (SEQ ID NO: 6), in the formula: Xaa7 is Ala, Val or Met. Preferably, Xaa7 is Ala.

[0065] Xaa8 is Thr or Leu. Preferably, Xaa8 is Thr.

[0066] Xaa9 is Gln or Gly. Preferably, Xaa9 is Gln.

[0067] Xaa10 is Arg or Lys. Preferably, Xaa10 is Arg.

[0068] Xaa11 is Leu.

[0069] Xaa12 is Ala or Ser. Preferably, Xaa12 is Ala.

[0070] Preferably, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7); Preferably, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, and Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7).

[0071] Xaa13 is any amino acid. Preferably, Xaa13 is Glu, Asp, Gln, Asn, or Lys, and more preferably, Xaa13 is Asp, Glu, or Gln.

[0072] Xaa14 is Glu, Asp or Phe. Preferably, Xaa14 is Glu.

[0073] Xaa15 is Leu.

[0074] Xaa16 is His.

[0075] Xaa17 is Lys or Arg. Preferably, Xaa17 is Lys.

[0076] Xaa18 is Leu.

[0077] Xaa19 is Gln or Lys. Preferably, Xaa19 is Gln.

[0078] Xaa20 is Thr.

[0079] Xaa21 is Tyr or Phe. Preferably, Xaa21 is Tyr.

[0080] Xaa22 is Pro.

[0081] Xaa23 is Arg or Lys. Preferably, Xaa23 is Arg.

[0082] Xaa24 is Thr or Lys. Preferably, Xaa24 is Thr.

[0083] Preferably, Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 8). Alternatively, Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 9).

[0084] Preferably, Xaa13 is Glu, Asp, Gln, Asn, or Lys (more preferably, Xaa13 is Asp, Glu, or Gln), and Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (sequence number 8).

[0085] Alternatively, Xaa13 is Glu, Asp, Gln, Asn, or Lys (more preferably, Xaa13 is Asp, Glu, or Gln), and Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 9).

[0086] Preferably, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), and Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 8).

[0087] Alternatively, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), and Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 9).

[0088] Preferably, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), and Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 8).

[0089] Alternatively, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), and Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 8).

[0090] Preferably, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, and Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (sequence number 8).

[0091] Alternatively, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, and Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (sequence number 9).

[0092] Preferably, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), and Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (sequence number 8).

[0093] Alternatively, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), and Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (sequence number 9).

[0094] Preferably, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7) and Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 8).

[0095] Alternatively, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7) and Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 9).

[0096] Preferably, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), and Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 8).

[0097] Alternatively, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), and Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 9).

[0098] Xaa25 is Pro, Lys or Asn. Preferably, Xaa25 is Pro. In alternative embodiments, it may be Lys.

[0099] Xaa26 is Val or Thr. Preferably, Xaa26 is Val. In alternative embodiments, it may be Thr.

[0100] Xaa27 is Gly or Val. Preferably, Xaa27 is Gly.

[0101] Xaa28 is Ser or Gly. Preferably, Xaa28 is Ser.

[0102] Preferably, Xaa27-Xaa28 are Gly-Ser.

[0103] Preferably, Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 8), or alternatively, Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 9) and Xaa27-Xaa28 are Gly-Ser.

[0104] Preferably, Xaa13 is Glu, Asp, Gln, Asn, or Lys (more preferably, Xaa13 is Asp, Glu, or Gln), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (sequence number 8), and Xaa27-Xaa28 are Gly-Ser.

[0105] Alternatively, Xaa13 is Glu, Asp, Gln, Asn, or Lys (more preferably, Xaa13 is Asp, Glu, or Gln), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (sequence number 9), and Xaa27-Xaa28 are Gly-Ser.

[0106] Preferably, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 8), and Xaa27-Xaa28 are Gly-Ser.

[0107] Alternatively, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 9), and Xaa27-Xaa28 are Gly-Ser.

[0108] Preferably, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 8), and Xaa27-Xaa28 are Gly-Ser.

[0109] Alternatively, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 9), and Xaa27-Xaa28 are Gly-Ser.

[0110] Preferably, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (sequence number 8), and Xaa27-Xaa28 are Gly-Ser.

[0111] Alternatively, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (sequence number 9), and Xaa27-Xaa28 are Gly-Ser.

[0112] Preferably, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (sequence number 8), and Xaa27-Xaa28 are Gly-Ser.

[0113] Alternatively, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (sequence number 9), and Xaa27-Xaa28 are Gly-Ser.

[0114] Preferably, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala, Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (sequence number 8), and Xaa27-Xaa28 are Gly-Ser.

[0115] Alternatively, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 9), and Xaa27-Xaa28 are Gly-Ser.

[0116] Preferably, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 8), and Xaa27-Xaa28 are Gly-Ser.

[0117] Alternatively, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 9), and Xaa27-Xaa28 are Gly-Ser.

[0118] Xaa29 is Lys, Asn, Gly, Ala, or Asp. Preferably, Xaa29 is Lys.

[0119] Xaa30 is Thr or Asn. Preferably, Xaa30 is Thr.

[0120] Xaa31 is any amino acid. Preferably, Xaa31 is Pro, Ala, Leu, Ile, Val, Ser, or Gln (most preferably Pro).

[0121] Xaa30-Xaa31 are preferably Thr-Pro.

[0122] Preferably, Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 8). Alternatively, Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 9), Xaa27-Xaa28 are Gly-Ser, Xaa30 is Thr, and Xaa31 is Pro, Ala, Leu, Ile, Val, Ser, or Gln (most preferably Pro).

[0123] Preferably, Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (sequence number 8), Xaa27-Xaa28 are Gly-Ser, Xaa30 is Thr, and Xaa31 is Pro, Ala, Leu, Ile, Val, Ser or Gln (most preferably Pro).

[0124] Alternatively, Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (sequence number 9), Xaa27-Xaa28 are Gly-Ser, Xaa30 is Thr, and Xaa31 is Pro, Ala, Leu, Ile, Val, Ser or Gln (most preferably Pro).

[0125] Preferably, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 8), Xaa27-Xaa28 are Gly-Ser, Xaa30 is Thr, and Xaa31 is Pro, Ala, Leu, Ile, Val, Ser or Gln (most preferably Pro).

[0126] Alternatively, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 9), Xaa27-Xaa28 are Gly-Ser, Xaa30 is Thr, and Xaa31 is Pro, Ala, Leu, Ile, Val, Ser or Gln (most preferably Pro).

[0127] Preferably, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), and Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa Xaa13 is Asp, Glu or Gln), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 8), Xaa27-Xaa28 are Gly-Ser, Xaa30 is Thr, and Xaa31 is Pro, Ala, Leu, Ile, Val, Ser or Gln (most preferably Pro).

[0128] Alternatively, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), and Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa1 Xaa13 is Asp, Glu or Gln, Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 9), Xaa27-Xaa28 are Gly-Ser, Xaa30 is Thr, and Xaa31 is Pro, Ala, Leu, Ile, Val, Ser or Gln (most preferably Pro).

[0129] Preferably, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (sequence number 8), Xaa27-Xaa28 are Gly-Ser, Xaa30 is Thr, and Xaa31 is Pro, Ala, Leu, Ile, Val, Ser or Gln (most preferably Pro).

[0130] Alternatively, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (sequence number 9), Xaa27-Xaa28 are Gly-Ser, Xaa30 is Thr, and Xaa31 is Pro, Ala, Leu, Ile, Val, Ser or Gln (most preferably Pro).

[0131] Preferably, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (sequence number 8), Xaa27-Xaa28 are Gly-Ser, Xaa30 is Thr, and Xaa31 is Pro, Ala, Leu, Ile, Val, Ser or Gln (most preferably Pro).

[0132] Alternatively, Xaa1 is absent, Xaa2 is Asn, Lys or Ser, Xaa3 is Thr, Xaa4 is Ala, Xaa5 is Thr, Xaa6 is Cys, Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (sequence number 9), Xaa27-Xaa28 are Gly-Ser, Xaa30 is Thr, and Xaa31 is Pro, Ala, Leu, Ile, Val, Ser or Gln (most preferably Pro).

[0133] Preferably, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 8), Xaa27-Xaa28 are Gly-Ser, Xaa30 is Thr, and Xaa31 is Pro, Ala, Leu, Ile, Val, Ser, or Gln (most preferably Pro).

[0134] Alternatively, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 9), Xaa27-Xaa28 are Gly-Ser, Xaa30 is Thr, and Xaa31 is Pro, Ala, Leu, Ile, Val, Ser, or Gln (most preferably Pro).

[0135] Preferably, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 8), Xaa27-Xaa28 are Gly-Ser, Xaa30 is Thr, and Xaa31 is Pro, Ala, Leu, Ile, Val, Ser or Gln (most preferably Pro).

[0136] Alternatively, Xaa7 to Xaa12 are Ala-Thr-Gln-Arg-Leu-Ala (SEQ ID NO: 7), Xaa13 is Glu, Asp, Gln, Asn or Lys (more preferably, Xaa13 is Asp, Glu or Gln), Xaa14 to Xaa24 are Glu-Leu-His-Lys-Leu-Lys-Thr-Tyr-Pro-Arg-Thr (SEQ ID NO: 9), Xaa27-Xaa28 are Gly-Ser, Xaa30 is Thr, and Xaa31 is Pro, Ala, Leu, Ile, Val, Ser or Gln (most preferably Pro).

[0137] Groups A and Q A is Q, Q-Ser, Q-Glu or Q-Lys. Preferably, A is Q, Q-Ser or Q-Lys.

[0138] Q is the formula: [ka] This is the part In the formula, R is C8 to C 28 It is an alkylene or alkenylene chain and R1 is -CO2H.

[0139] Within the Q portion of the compound, the R group is an alkylene or alkenylene chain that is linked at one end via an amide bond to the B portion of the molecule (or to a Ser residue in A if A is Q-Ser, or to a Lys residue in A if A is Q-Lys) by a Glu residue in the Q portion of the molecule. At its other end, the R alkylene or alkenylene chain is linked to an R acid group (COH).

[0140] Option (i) above describes the situation where the Glu residue of Q is attached to the B portion of the compound (or to the Ser residue of A if A is Q-Ser, or to the Lys residue of A if A is Q-Lys) via the γ-carboxylic acid group of Glu. Option (ii) above describes the situation where the Glu residue of the Q portion of the compound is attached to the B portion of the compound (or to the Ser residue of A if A is Q-Ser, or to the Lys residue of A if A is Q-Lys) via the α-carboxylic acid group of Glu. In a preferred embodiment of the invention, the Q portion of the compound is of formula (I), i.e., the Glu residue of the Q portion is attached to the rest of the compound via the γ-carboxylic acid group of the Glu residue moiety of Q.

[0141] Generally, R has an even number of carbon atoms. For example, R can be an alkylene or alkenylene chain found in naturally occurring fatty acids. The root fatty acid has a chain length that is two carbon atoms longer than the number of carbon atoms in the R alkylene or alkenylene chain.

[0142] R is preferably C 16 ~C 18 Preferably, R is a straight chain alkylene or alkenylene group. For example, R is preferably a straight chain alkylene or alkenylene group. For example, R is C 16 or C 18It can be a straight chain alkylene group. For example, R can be C 16 This can be provided by an octadecanedioic acid moiety, for example, when R is C 18 In the case of the group, this may be provided by an eicosanedioic acid moiety.

[0143] In a preferred embodiment, R is C 18 It is an alkylene group.

[0144] In the table of Figure 1, the specific identity of the dicarboxylic acid portion of Q is indicated by reference to the root aliphatic dicarboxylic acid that forms part of Q. That is, when "eicosanedioic acid" appears in the table of Figure 1, R is C 18 It is a straight chain alkylene group. The glutamic acid portion of Q is identified by the symbol "γGlu" or "Glu." "γGlu" refers to the Q portion of a compound according to formula (I). "Glu" refers to the Q portion of a compound according to formula (II).

[0145] In particularly preferred embodiments, the compound of the invention is one of the specific compounds of the invention set forth in the table of FIG.

[0146] Disulfide bridges In all embodiments of the invention, compounds of formula ABC have a Cys residue near the N-terminus of the peptide sequence as shown, and five (or sometimes six) additional Cys residues along the peptide portion of the molecule toward the C-terminus. The Cys residues contain a thiol group side chain that can be oxidized to form a disulfide -SS- bond between two Cys residues. While non-disulfide bonded versions of the compounds of the invention are valuable (e.g., as intermediate compounds in the generation of disulfide bonded versions of the compounds of the invention), the presence of this -SS- bond is essential for biological activity and is therefore a preferred feature according to certain embodiments of the invention. All of the compounds listed in the table of Figure 1 were made with a -SS- bond between the two Cys residues.

[0147] Derivatives and Salts The present invention provides compounds of formula ABC, derivatives of such compounds and salts or solvates of such compounds and derivatives.

[0148] The compounds, derivatives and salts may be produced by recombinant methods well known in the art, or may be produced by synthetic methods, again well known in the art.

[0149] derivative In some embodiments, the present invention relates directly to compounds of formula ABC, but not derivatives, while in other embodiments, the present invention relates to derivatives of compounds of formula ABC. Derivatives can include, for example, one or more derivatizations selected from amidation, glycosylation, carbamylation, acylation, sulfation, phosphorylation, further cyclization, further lipidation, pegylation, and fusion to another peptide or protein to form a fusion protein. For example, derivatives can include one or more derivatizations selected from amidation, glycosylation, carbamylation, acylation, sulfation, phosphorylation, cyclization, lipidation, and pegylation. The structure can be modified at random positions within the molecule or at predetermined positions within the molecule and can include one, two, three, or more attached chemical moieties. According to certain embodiments, the term "derivative" does not extend to compounds in which the peptide sequence has been altered.

[0150] A derivative can be, for example, a fusion protein in which the ABC structure of a compound of the invention is fused to another protein or polypeptide (fusion partner) using recombinant methods known in the art. Alternatively, such fusion proteins can be synthetically synthesized by any known method. Such a fusion protein comprises a structure of formula ABC. Any suitable peptide or protein can be used as the fusion partner (e.g., serum albumin, carbonic anhydrase, glutathione-S-transferase, thioredoxin, etc.). Such a fusion protein can be made by linking the carboxy terminus of the fusion partner to the amino terminus of the structure of formula ABC (or vice versa). Optionally, a cleavable linker can be used to link the structure of formula ABC to the fusion partner. The resulting cleavable fusion protein can be cleaved in vivo to liberate the active form of the compound of the invention. Examples of such cleavable linkers include, but are not limited to, the linkers Asp-Asp-Asp-Asp-Tyr (SEQ ID NO: 10), Gly-Pro-Arg, Ala-Gly-Gly, and His-Pro-Phe-His-Leu (SEQ ID NO: 11), which can be cleaved by enterokinase, thrombin, ubiquitin cleavage enzyme, and renin, respectively. For further details, see, e.g., U.S. Patent No. 6,410,707, the contents of which are incorporated herein by reference.

[0151] The derivatives of the present invention may be, for example, physiologically functional derivatives of the structure of formula ABC. The term "physiologically functional derivative" is used herein to refer to a chemical derivative of a compound of formula ABC that has the same physiological function as the corresponding unmodified compound. For example, a physiologically functional derivative may be convertible in vivo to a compound of formula ABC. According to the present invention, examples of physiologically functional derivatives include esters, amides, and carbamates; preferably, esters and amides.

[0152] For example, pharmaceutically acceptable esters and amides of the compounds of the present invention can comprise a C1-20 alkyl-, C2-20 alkenyl-, C5-10 aryl-, C5-10ar-C1-20 alkyl-, or amino acid-ester or amide group attached to an appropriate moiety, formed, for example, by the reaction of an alkyl, alkenyl, aryl, aralkyl, or aminoalkyl group containing an alcohol or amino moiety with an acid moiety present in a compound of formula ABC, or by the reaction of an alkyl, alkenyl, aryl, aralkyl, or aminoalkyl group containing an activated acyl group with an alcohol or amine group present in a compound of formula ABC. Examples of suitable moieties are hydrophobic substituents having 4 to 26 carbon atoms, preferably 5 to 19 carbon atoms. Suitable lipid groups include fatty acids (e.g., lauroyl (C 12 H 23 ), Palmityl (C 15 H 31 ), Oleil (C 15 H 29 ) or stearyl (C 17 H 35 )) and bile acids (e.g., cholic acid or deoxycholic acid).

[0153] Lipidation (both with respect to the compounds of the present invention and their derivatives) significantly increases absorption of the compounds and prolongs blood and tissue retention of the compounds compared to the absorption rate of the corresponding non-lipidated compounds. Suitable lipid-containing moieties are hydrophobic substituents having 4 to 26 carbon atoms, preferably 5 to 19 carbon atoms. Suitable lipid groups include fatty acids (e.g., lauroyl (C 12 H 23 ), Palmityl (C 15 H 31 ), Oleil (C 15 H 29 ) or stearyl (C 17 H 35)) and bile acids (e.g., cholic acid (cholate) or deoxycholic acid). While the lipid-functionalized compounds of the present invention may have benefits in certain circumstances, it is expected that in most cases it will be preferred if the compounds of the present invention are not further derivatized so that there are no additional lipid groups present beyond the aliphatic dicarboxylic acid portion of the compound according to formula ABC.

[0154] Cyclization methods (in the context of both the compounds of the present invention and derivatives of those compounds) include cyclization via disulfide bridge formation and head-to-tail cyclization using a cyclization resin. Cyclized peptides may have enhanced stability, including increased resistance to enzymatic degradation, as a result of their conformational constraints. Cyclization may be particularly advantageous when the uncyclized peptide contains an N-terminal cysteine ​​group. Suitable cyclized peptides include monomeric and dimeric head-to-tail cyclized structures. Cyclized peptides may contain one or more additional residues, particularly additional cysteines incorporated for the purposes of disulfide bond formation or side chains incorporated for the purposes of resin-based cyclization. While further cyclization may be beneficial in certain circumstances, it is expected that in most cases it will be preferable if the compounds of the present invention are not further cyclized, so that there is no further cyclization beyond that achieved by -SS- between the two cysteine ​​residues in compounds of formula ABC.

[0155] The derivative can be, for example, a pegylated structure of formula ABC. Pegylated compound derivatives of the invention can provide advantages such as increased solubility, stability, and circulation time of the polypeptide, or reduced immunogenicity (see U.S. Pat. No. 4,179,337, the contents of which are incorporated herein by reference).

[0156] Chemical moieties for derivatization of the compounds of the invention can also be selected from water-soluble polymers, such as polyethylene glycol, ethylene glycol / propylene glycol copolymers, carboxymethylcellulose, dextran, polyvinyl alcohol, and the like. Polymer moieties for derivatization of the compounds of the invention can be of any molecular weight and can be branched or unbranched. For ease of handling and manufacturing, the preferred molecular weight of polyethylene glycol for derivatization of the compounds of the invention is about 1 kDa to about 100 kDa, the term "about" indicating that in polyethylene glycol preparations, some molecules will be heavier and some will be lighter than the stated molecular weight. Polymers of other molecular weights can be used depending on the desired therapeutic profile, e.g., the desired duration of sustained release, the effect, if any, on biological activity, ease of handling, the degree or lack of antigenicity, and other known effects of polyethylene glycol on therapeutic proteins or analogs. For example, polyethylene glycol may be used in the following ranges: about 200, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000, 11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15 ,500, 16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500, 20,000, 25,000, 30,000, 35,000, 40,000, 45,000, 50,000, 55,000, 60,000, 65,000, 70,000, 75,000, 80,000, 85,000, 90,000, 95,000 or 100,000 kDa.

[0157] salt Salt forms of the compounds of the present invention and derivatives of such compounds also form part of the present invention. In some embodiments, the salts are salts of compounds of the present invention. In other embodiments, the salts are salts of derivatives of compounds of the present invention.

[0158] Salts of the compounds of the present invention include salts that are pharmaceutically acceptable, i.e., suitable for use in pharmaceutical products. However, salts having pharmaceutically unacceptable counterions are also within the scope of the invention for use as intermediates in the preparation of the compounds.

[0159] Suitable salts according to the present invention include those formed with organic or inorganic acids or bases. Pharmaceutically acceptable acid addition salts include those formed with hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, citric acid, tartaric acid, acetic acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, trifluoroacetic acid, succinic acid, perchloric acid, fumaric acid, maleic acid, glycolic acid, salicylic acid, oxaloacetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, formic acid, benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and isethionic acid. Other acids, such as oxalic acid, may be useful as intermediates in obtaining the final compound of the present invention.

[0160] Pharmaceutically acceptable salts with bases include ammonium salts, alkali metal salts such as potassium salts and sodium salts, alkaline earth metal salts such as calcium salts and magnesium salts, and salts with organic bases such as dicyclohexylamine and N-methyl-D-glucamine.

[0161] solvate Those skilled in the art of organic chemistry and / or medicinal chemistry will recognize that many organic compounds can form complexes with solvents in which they are reacted or from which they are precipitated or crystallized. Such complexes are known as "solvates." For example, a complex with water is known as a "hydrate." The present invention also encompasses solvates of the compounds of the invention, solvates of derivatives of the compounds, and solvates of salts of the derivatives.

[0162] Those skilled in the art of organic chemistry and / or medicinal chemistry will also recognize that many organic compounds can exist in different forms, for example, as amorphous material and / or in one or more crystalline forms. Different physical forms of an organic compound are known as polymorphs. The present invention encompasses all such different physical forms of the compounds of the present invention, as well as different physical forms of derivatives and salts.

[0163] biological activity Amylin appears to have at least three different receptors that can bind with high affinity, including the calcitonin receptor along with one of at least three altered receptor activity-modifying proteins (RAMPs)—RAMP1, RAMP2, or RAMP3 (Hay DL, Christopoulos G, Christopoulos A, Sexton PM (2004). Biochem. Soc. Trans. 32(Part 5):865-7).

[0164] Compounds of the present invention that have activity at human amylin receptors can be considered amylin receptor agonists. This can be assessed, for example, by in vitro or cell binding assays, or by reporter assays. Preferred compounds of the present invention exhibit activity at human amylin receptors that is at least 50 times less than that of human amylin, preferably at least 30, 20, 10, 5, 3 or 2 times less than that of human amylin, when tested, for example, according to the assays described in the Examples below.

[0165] The method for evaluating the activity of amylin receptor is well known. For example, homogeneous time resolved fluorescence (HFT) (Change CL et al., Activation of Calcitonin Receptor and Calcitonin Receptor-like Receptor by Membrane-anchored Ligands, Journal of Biological Chemistry, Vol. 285, No. 2, January 8, 2010, pp. 1075-1080) discloses a method for assaying amylin receptor activation. A specific method is described in one of the examples herein.

[0166] The compounds of the present invention meet some, or more preferably all, of the following criteria: 1) sustained biological activity at human amylin receptors, resulting in appetite suppression; 2) High solubility in aqueous solution at pH 3.8, allowing an effective dose to be administered with a low volume injection (thereby reducing the pain of the injection). Solubility can be easily assessed by simple in vitro tests; 3) long-term activity in vivo (when assessed in humans or animal models) to allow injections no more frequently than once per day, preferably no more than twice per week, or more preferably no more than once per week, while still providing acceptable therapeutic or cosmetic benefit; 4) Good weight loss (when assessed in human subjects or animal models).

[0167] According to certain embodiments of the invention, particularly those relating to weight loss, obesity, carbohydrate metabolism and diabetes, the compounds, derivatives and salts of the invention have one or both of the following features: A. Sufficient solubility between pH 2 and pH 8 (e.g., between pH 4 and 8 or between pH 4 and pH 7) to allow an effective dose to be administered in a volume of less than 1 ml, less than 0.5 ml, or less than 0.3 ml; B. One, some, or all of the additional features 1-6 listed above.

[0168] Pharmacokinetics, duration of action and solubility The compounds of the present invention exhibit potent and long-lasting action in vivo after subcutaneous administration. To achieve this, the compounds are required to have both good activity at the biological target and excellent pharmacokinetic properties.

[0169] The compounds of the present invention have a therapeutically useful duration of action, which is manifested in the beneficial effects observed over several days in the experiments described herein below. The half-life of the compounds of the present invention can be evaluated using any suitable pharmacokinetic model, for example, a porcine PK model. Preferred compounds of the present invention have been found to have a significantly longer half-life than human amylin. In addition to exhibiting a long in vivo half-life, the compounds of the present invention have good storage stability, with no significant degradation observed upon storage in solution (e.g., sterile water or sterile buffer solution) at 4°C for 4 weeks.

[0170] Poor water solubility is a known problem for lipid-containing molecules. In contrast, the compounds of the present invention have very good solubility.

[0171] situation The present invention also provides a compound, derivative or salt of the invention or a composition comprising the compound, derivative or salt together with a pharmaceutically acceptable carrier, and optionally a further therapeutic agent, for use as a pharmaceutical.

[0172] The present invention also provides a method of treating or preventing a disease or disorder or other unwanted physiological condition in a subject, comprising administering a therapeutically effective amount of a compound, derivative, or salt of the invention, or a composition comprising the compound, derivative, or salt, together with a pharmaceutically acceptable carrier and, optionally, a further therapeutic agent. Preferably, the compound, derivative, salt, or composition is administered subcutaneously.

[0173] According to certain embodiments, the disease or disorder or other unwanted physiological condition is diabetes or obesity, in particular diabetes (eg, type II diabetes).

[0174] According to certain embodiments, the disease or disorder or other undesired physiological condition may be the physiological condition of being overweight.

[0175] The subject to which the compound is administered may be overweight, for example, obese. Alternatively, or in addition, the subject may be a diabetic patient, for example, with insulin resistance or glucose intolerance or both. The subject may have diabetes mellitus, for example, the subject may have type II diabetes. The subject may be overweight, for example, obese, and may have diabetes mellitus, for example, type II diabetes.

[0176] Additionally or alternatively, the subject may have or be at risk of having the disorder that obesity or overweight is a risk factor.Such disorders include but are not limited to heart disease, cardiovascular disease, such as hypertension, atherosclerosis, congestive heart failure and dyslipidemia; stroke; gallbladder disease; osteoarthritis; sleep apnea; reproductive disorders, such as polycystic ovarian syndrome; cancer, such as breast cancer, prostate cancer, colon cancer, endometrial cancer, kidney cancer and esophageal cancer; varicose veins; acanthosis nigricans; eczema; exercise intolerance; insulin resistance; hypertension, hypercholesterolemia; cholelithiasis; osteoarthritis; orthopedic injury; insulin resistance, such as type 2 diabetes and syndrome X; and thromboembolic disease (see Kopelman, Nature 404:635-43, 2000; Rissanen et al., British Med.J.301, 835, 1990).

[0177] Other disorders associated with obesity include depression, anxiety, panic attacks, migraines, headaches, PMS, chronic pain conditions, fibromyalgia, insomnia, impulsivity, obsessive-compulsive disorder, and myoclonus. Certain neurological disorders and certain forms of neurodegeneration are also associated with obesity. Furthermore, obesity is a recognized risk factor for increased incidence of complications of general anesthesia (see, for example, Kopelman, Nature 404:635-43, 2000). In general, obesity carries a significant risk of comorbidities such as those listed above.

[0178] Other diseases or disorders associated with obesity include maternal obesity, which is associated with an increased incidence of birth defects, neural tube defects, and carpal tunnel syndrome (CTS); chronic venous insufficiency (CVI); daytime sleepiness; deep vein thrombosis (DVT); end-stage renal disease (ESRD); gout; fever disorders; impaired immune response; respiratory dysfunction; infertility; liver disease; back pain; obstetric and gynecological complications; pancreatitis; and abdominal hernia; acanthosis nigricans; endocrine disorders; chronic hypoxia and hypercapnia; dermatological effects; These include elephantiasis; gastroesophageal reflux; heel spurs; leg edema; mammegaly causing significant problems such as bra strap pain, skin breakdown, neck pain, chronic odor and infection in the skin folds under the breasts; large anterior abdominal wall masses, such as abdominal panniculitis with frequent panniculitis, that interfere with walking and cause frequent infections, odor, difficulty dressing, and back pain; musculoskeletal disorders; pseudotumor cerebri (or benign intracranial hypertension), and sliding hiatal hernia.

[0179] In some embodiments, the disease or disorder can be non-alcoholic fatty liver disease.

[0180] According to certain embodiments, the disease or disorder or other undesired physiological condition may be an undesired weight, despite not being obese or overweight. The subject may be of normal weight (including, but not limited to, subjects who were previously overweight or obese, and subjects who wish to prevent a return to an unhealthy weight). The subject may be a subject who desires weight loss, for example, a female and / or male subject who desires a change in appearance. In some cases where the subject is of normal weight, aspects of the invention may relate to cosmetic treatments rather than therapeutic treatments.

[0181] The present invention also provides methods for reducing appetite in a subject, reducing food intake in a subject, reducing calorie intake in a subject, improving insulin release in a subject, improving carbohydrate metabolism in a subject, and / or improving carbohydrate tolerance in a subject, comprising administering a therapeutically effective amount of a compound, derivative, salt, or composition of the invention. Such methods may involve treating a subject with a pre-diabetic condition, such as insulin insensitivity or pre-diabetes.

[0182] The present invention also provides a method for improving the lipid profile in a subject, comprising administering a therapeutically effective amount of a compound, derivative, salt or composition of the present invention.The present invention also provides a method for alleviating a condition or disorder that can be alleviated by reducing nutrient availability, comprising administering a therapeutically effective amount of a compound, derivative, salt or composition of the present invention.

[0183] The compounds, derivatives, salts or compositions of the invention can be used to control and treat body weight, for example, to reduce or prevent obesity, and in particular to prevent and reduce weight gain, induce and promote weight loss, and reduce obesity as measured by body mass index. The compounds, derivatives, salts or compositions of the invention can be used to maintain any one or more of a desired body weight, a desired body mass index, a desired appearance, and good health.

[0184] The present invention can also be used to treat, prevent, ameliorate, or alleviate conditions or disorders caused by, accompanied by, or exacerbated by relatively high nutrient availability. The term "condition or disorder that can be alleviated by reducing calorie (or nutrient) availability" is used herein to refer to any condition or disorder in a subject that is either caused by, accompanied by, or exacerbated by relatively high nutrient availability, or that can be alleviated by reducing nutrient availability, for example, by reducing food intake. Subjects who are insulin resistant, glucose intolerant, or have any form of diabetes mellitus, such as type 1 diabetes, type 2 diabetes, or gestational diabetes, can also benefit from the method according to the present invention.

[0185] Conditions or disorders associated with increased caloric intake include, but are not limited to, insulin resistance, glucose intolerance, obesity, diabetes, including type 2 diabetes, eating disorders, insulin resistance syndrome, and Alzheimer's disease.

[0186] The present invention also provides a compound, derivative, salt or composition of the invention for use in the treatment of obesity or diabetes.

[0187] The invention also provides a compound, derivative, salt or composition of the invention for use in increasing energy expenditure in a subject, improving insulin release in a subject, improving carbohydrate tolerance in a subject and / or improving carbohydrate metabolism in a subject. Such use may relate to the treatment of a subject with a pre-diabetic condition, such as insulin insensitivity or pre-diabetes.

[0188] The invention also provides a compound, derivative, salt or composition of the invention for use in reducing appetite in a subject, in reducing food intake in a subject, in reducing calorie intake in a subject, in improving insulin release in a subject, and / or in improving carbohydrate tolerance in a subject. Such uses may relate to the treatment of a subject with a pre-diabetic condition, such as insulin insensitivity or pre-diabetes.

[0189] The invention also provides the use of a compound, derivative, salt or composition of the invention for the manufacture of a medicament for the treatment of obesity or diabetes in a subject, which may be as described above in relation to other aspects of the invention.

[0190] The present invention also provides the use of a compound, derivative or salt of the invention for the manufacture of a medicament for improving insulin release in a subject, for improving carbohydrate tolerance in a subject and / or for improving carbohydrate metabolism in a subject. Such use may relate to the treatment of a subject with a pre-diabetic condition, such as insulin insensitivity or pre-diabetes.

[0191] The invention also provides the use of a compound, derivative or salt of the invention for the manufacture of a medicament for use in reducing appetite in a subject, reducing food intake in a subject, reducing calorie intake in a subject, improving insulin release in a subject, and / or improving carbohydrate tolerance in a subject.

[0192] In certain embodiments, the compounds, derivatives, salts, or compositions of the present invention are administered parenterally. In other embodiments, the compounds, derivatives, salts, or compositions of the present invention are administered subcutaneously, intravenously, intramuscularly, intranasally, transdermally, or sublingually. In other embodiments, the compounds, derivatives, salts, or compositions of the present invention are administered orally. In a preferred embodiment, the compounds, derivatives, salts, or compositions of the present invention are administered subcutaneously.

[0193] The compounds, derivatives, salts, or compositions of the present invention are preferably used to treat human subjects. However, although the compounds, derivatives, salts, or compositions of the present invention are typically used to treat human subjects, they can also be used to treat similar or identical conditions in other vertebrates, such as other primates; livestock, such as pigs, cattle, and poultry; sport animals, such as horses, or companion animals, such as dogs and cats.

[0194] composition The compounds of the present invention or their derivatives and / or salts are preferably present in pharmaceutical formulations or compositions. Accordingly, the present invention provides compositions comprising the compounds, derivatives, or salts of the present invention together with a pharmaceutically acceptable excipient, and optionally another therapeutic ingredient. The compositions comprising the compounds, derivatives, or salts are suitable for pharmaceutical use. According to certain preferred embodiments, the compositions are present in a syringe or other administration device for subcutaneous administration to humans. According to certain preferred embodiments, the compositions have a pH of less than 5. The compositions of the present invention can take the pharmaceutical formulation forms described below.

[0195] Pharmaceutical formulations according to the present invention include those suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous and intraarticular), inhalation (including fine particle dusts or mists which may be generated by various types of metered dose pressurized aerosols, nebulizers or insufflators), rectal and topical (including cutaneous, transdermal, transmucosal, buccal, sublingual and ocular) administration, although the most suitable route may depend, for example, on the condition and disorder of the recipient.

[0196] Preparations can be conveniently provided in unit dosage form and can be prepared by any method well known in the field of pharmacy.All methods include the step of mixing active ingredient with carrier that constitutes one or more accessory ingredients.Generally, preparations are prepared by uniformly and intimately mixing active ingredient with liquid carrier or finely divided solid carrier or both, and then, if necessary, shaping product into desired preparation form.

[0197] Formulations of the present invention suitable for oral administration can be presented as discrete units such as capsules, cachets, or tablets, each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or suspension in an aqueous or non-aqueous liquid; or as an oil-in-water or water-in-oil liquid emulsion. The active ingredient can also be presented as a bolus, electuary, or paste. Various pharmaceutically acceptable carriers and their formulations are described in standard formulation treatises, such as Remington's Pharmaceutical Sciences by E.W. Martin. See also Wang, Y.J. and Hanson, M.A., Journal of Parenteral Science and Technology, Technical Report No. 10, Suppl. 42:2S, 1988, the contents of which are incorporated herein by reference.

[0198] Tablets can be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing the active ingredient in a free-flowing form, such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, glidant, surface active agent, or dispersing agent in a suitable machine. Molded tablets can be made by molding a mixture of powdered compound moistened with an inert liquid diluent in a suitable machine. Tablets can optionally be coated or scored and can be formulated to provide sustained or controlled release of the active ingredient therein. The compound can be administered, for example, in a form suitable for immediate release or sustained release. Immediate release or sustained release can be achieved by using a suitable pharmaceutical composition containing the compound, or, particularly in the case of sustained release, by using a device such as a subcutaneous implant or osmotic pump. The compound can also be administered liposomally.

[0199] Preferably, the composition according to the present invention is suitable for subcutaneous administration, for example, by injection. According to certain embodiments, the composition may contain metal ions, such as copper ions, iron ions, aluminum ions, zinc ions, nickel ions, or cobalt ions. The presence of such ions can limit solubility, thereby delaying absorption from the subcutaneous administration site into the circulatory system.

[0200] Exemplary compositions for oral administration include suspending agents, which may contain, for example, microcrystalline cellulose for bulking, alginic acid or sodium alginate as a suspending agent, methylcellulose as a thickener, and sweeteners or flavoring agents, such as those known in the art; and immediate-release tablets, which may contain, for example, microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate, and / or lactose, and / or other excipients, binders, fillers, disintegrants, diluents, and lubricants, such as those known in the art. Such compositions may also contain penetration enhancers. The compounds of the present invention can also be delivered through the oral cavity by sublingual and / or buccal administration. Molded tablets, compressed tablets, or freeze-dried tablets are exemplary forms that can be used. Exemplary compositions include those in which the compounds are formulated with fast-dissolving diluents, such as mannitol, lactose, sucrose, and / or cyclodextrins. High molecular weight excipients, such as cellulose (avicel) or polyethylene glycol (PEG), may also be included in such formulations. Such formulations may also include excipients to aid mucoadhesion, such as hydroxypropyl cellulose (HPC), hydroxypropyl methylcellulose (HPMC), sodium carboxymethyl cellulose (SCMC), maleic anhydride copolymers (e.g., Gantrez), and agents to control release, such as polyacrylic copolymers (e.g., Carbopol 934). Lubricants, glidants, flavors, colorants, and stabilizers may also be added for ease of manufacture and use.

[0201] Formulations for parenteral administration include aqueous and non-aqueous sterile injection solutions, which may contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions, which may contain suspending agents and thickening agents. The formulations may be presented in unit-dose or multi-dose containers, such as sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) state, requiring only the addition of a sterile liquid carrier, such as saline or water for injection, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets of the type described above. Exemplary compositions for parenteral administration include injectable solutions or suspensions, which may contain, for example, a suitable non-toxic parenterally acceptable diluent or solvent, such as mannitol, 1,3-butanediol, water, Ringer's solution, isotonic sodium chloride solution, or other suitable dispersing or wetting agents and suspending agents, such as synthetic mono- or diglycerides and fatty acids, such as oleic acid or cremaphor. The aqueous carrier may be, for example, an isotonic buffer solution having a pH of about 3.0 to about 8.0, preferably about 3.5 to about 7.4, e.g., 3.5 to 6.0, e.g., 3.5 to about 5.0. Useful buffers include sodium citrate-citric acid, sodium phosphate-phosphate, and sodium acetate / acetic acid buffers. The composition preferably does not contain any compounds known to be harmful to peptide compounds.

[0202] Excipients that can be included are, for example, other proteins, such as human serum albumin or plasma preparations. If desired, the pharmaceutical compositions can also contain minor amounts of nontoxic auxiliary substances, such as wetting or emulsifying agents, preservatives, and pH buffering agents, such as, for example, sodium acetate or sorbitan monolaurate.

[0203] Exemplary compositions for nasal aerosol or inhalation administration include solutions in saline, which may contain, for example, benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, and / or other solubilizing or dispersing agents, such as those known in the art. Conveniently, in compositions for nasal aerosol or inhalation administration, the compounds of the present invention are delivered in the form of an aerosol spray presentation from a pressurized pack or nebulizer using a suitable propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. In the case of pressurized aerosols, the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges, e.g., of gelatin, for use in inhalers or insufflators can be formulated to contain a powder mix of the compound and a suitable powder base, e.g., lactose or starch. In one specific, non-limiting example, the compounds of the present invention are administered as an aerosol from a metered valve via an aerosol adapter, also known as an actuator. Optionally, a stabilizer is also included, and / or porous particles are included for deep lung delivery (see, eg, US Pat. No. 6,447,743).

[0204] Formulations for rectal administration can be presented as retention enemas or suppositories with conventional carriers such as cocoa butter, synthetic glyceride esters, or polyethylene glycols. Such carriers are typically solid at ordinary temperatures but liquefy and / or melt in the rectal cavity to release the drug.

[0205] Formulations for topical administration in the oral cavity, e.g., buccal or sublingual, include, for example, lozenges comprising the active ingredient in a flavored base such as sucrose and acacia or tragacanth, and pastilles comprising the active ingredient in a base such as gelatin and glycerin or sucrose and acacia. Exemplary compositions for topical administration include a topical carrier such as Plastibase (mineral oil gelled with polyethylene).

[0206] Preferred unit dosage formulations are those containing an effective dose, as herein above recited, or an appropriate fraction thereof, of the active ingredient.

[0207] It will be understood that in addition to the ingredients particularly mentioned above, the formulations of the present invention may include other agents conventional in the art having regard to the type of formulation being discussed; for example, those suitable for oral administration may include flavoring agents.

[0208] The compounds, derivatives, and salts of the present invention can also be suitably administered as sustained-release systems. Suitable examples of sustained-release systems of the present invention include suitable polymeric materials, such as semipermeable polymer matrices in the form of shaped articles, e.g., films or microcapsules; suitable hydrophobic materials, e.g., emulsions in acceptable oils; or ion exchange resins; and sparingly soluble derivatives of the compounds of the present invention, e.g., sparingly soluble salts. The sustained-release systems can be administered orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, or topically, for example, as powders, ointments, gels, drops, or transdermal patches; bucally; or as oral or nasal sprays.

[0209] The preparation for administration can be suitably formulated to provide controlled release of the compounds, derivatives and salts of the present invention. For example, the pharmaceutical composition can be in the form of particles containing one or more of biodegradable polymers, polysaccharide gelling and / or bioadhesive polymers, amphiphilic polymers, and agents that can modify the interfacial properties of particles of the compounds of the present invention. These compositions exhibit certain biocompatible features that allow for controlled release of the active substance. See U.S. Patent No. 5,700,486, the contents of which are incorporated herein by reference.

[0210] The use of controlled-release compositions is preferred for indications such as the treatment of obesity and / or diabetes, where it is desirable to maximize the time between injections. However, for indications such as providing neuroprotection or cardioprotection (e.g., after a suspected myocardial infarction or stroke), where it is desirable to achieve a therapeutic plasma concentration of the active agent in as short a time as possible, an immediate-release formulation is preferred. In such cases, a dosing regimen involving administration of a dose of an immediate-release formulation of the active agent (i.e., as soon as possible after a suspected myocardial infarction or stroke) and subsequent administration of a dose of a controlled-release formulation of the active agent may be preferred.

[0211] The compounds, derivatives, or salts of the invention can be delivered via pump (see Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng. 14:201, 1987; Buchwald et al., Surgery 88:507, 1980; Saudek et al., N Engl. J. Med. 321:574, 1989) or by continuous subcutaneous infusion, for example, using a minipump. An intravenous bag solution can also be used. An important factor in selecting the appropriate dose is the results obtained, as measured by total body weight loss or fat to lean mass ratio, or other criteria deemed appropriate by the physician for measuring the control or prevention of obesity or obesity-related conditions. Other controlled-release systems are discussed in the review by Langer (Science 249:1527-1533, 1990), incorporated herein by reference. In another aspect of the disclosure, the compounds of the present invention are delivered via an implantable pump, for example, as described in U.S. Pat. No. 6,436,091; U.S. Pat. No. 5,939,380; U.S. Pat. No. 5,993,414, the contents of which are incorporated herein by reference.

[0212] Implantable drug infusion devices are used to provide patients with a constant and long-term dosage or infusion of a drug or any other therapeutic agent. Essentially, such devices can be categorized as either active or passive. The compounds, derivatives, or salts of the present invention can be formulated as depot preparations. Such long-acting depot preparations can be administered, for example, by subcutaneous or intramuscular implantation; or by intramuscular injection. Thus, for example, the active ingredient can be formulated with a suitable polymeric or hydrophobic material, for example, as an emulsion in an acceptable oil; or with an ion exchange resin; or as a sparingly soluble derivative, for example, as a sparingly soluble salt.

[0213] The therapeutically effective amount of the active agent of the present invention can be administered as a single pulse dose, as a bolus dose, or as a pulse dose administered over time.Therefore, in pulse dose administration, a bolus dose of active agent is provided, followed by a period in which no active agent is administered to the subject, and then a second bolus dose is administered.In a specific non-limiting example, pulse dose is administered for one day, one week, or one month.

[0214] Combination therapy In certain embodiments, a therapeutically effective amount of a compound, derivative, salt, or composition of the present invention is administered together with a therapeutically effective amount of one or more additional therapeutic agents. The compound, derivative, or salt can, for example, be administered simultaneously with one or more additional therapeutic agents, or can be administered sequentially or separately. Thus, the present invention provides a compound, derivative, or salt of the present invention for use as a pharmaceutical, wherein the compound, derivative, or salt is for use together with a therapeutically effective amount of one or more additional therapeutic agents (e.g., for simultaneous, sequential, or separate administration). In certain embodiments, an active agent of the present invention is formulated and administered together with one or more additional therapeutic agents as a single dose.

[0215] In certain embodiments, the additional therapeutic agent(s) is an additional antidiabetic agent, appetite suppressant, food intake reducer, plasma glucose lowering agent, or plasma lipid altering agent. Specific non-limiting examples of additional appetite suppressants include caglilintide, pramlintide, amfepramone (diethylpropion), phentermine, mazindol, and phenylpropanolamine, fenfluramine, dexfenfluramine, phendimetrazine, benzphetamine, sibutramine, rimonabant, topiramate, fluoxetine, bupropion, zonisamide, naltrexone, orlistat, and cetilistat. Specific non-limiting examples of additional antidiabetic agents include metformin, phenformin, rosiglitazone, pioglitazone, troglitazone, repaglinide, nateglinide, tolbutamide, acetohexamide, tolazamide, chlorpropamide, glipizide, glyburide, glimepiride, gliclazide, fibroblast growth factor 21, miglitol, acarbose, exenatide, pramlintide, vildagliptin, and sitagliptin.

[0216] In a preferred embodiment, the additional therapeutic agent or agents is GLP-1 or a GLP-1 derivative, analogue or agonist.

[0217] In an alternative embodiment, the additional therapeutic agent or agents is insulin or an insulin analog, derivative or agonist.

[0218] In an alternative embodiment, the additional therapeutic agent or agents is GIP or a GIP analog, derivative or agonist.

[0219] In an alternative embodiment, the additional therapeutic agent or agents is glucagon or a glucagon analogue, derivative or agonist.

[0220] Dosage The compounds, derivatives, salts or compositions of the invention can be administered whenever an effect, such as appetite suppression, reduced food intake or reduced calorie intake, is desired or shortly before whenever the effect is desired, for example, but not limited to, about 10 minutes, about 15 minutes, about 30 minutes, about 60 minutes, about 90 minutes or about 120 minutes before the time the effect is desired.

[0221] Alternatively, or in addition, the compounds, derivatives, salts or compositions of the invention may be administered in a conventional dosing regimen, for example, once daily, once weekly, once every 10 days or once every two weeks.

[0222] The therapeutically effective amount of an active agent of the present invention depends on the molecule utilized, the subject being treated, the severity and type of affliction, and the mode and route of administration. For example, a therapeutically effective amount of a compound of the present invention can vary from about 0.01 μg / kilogram (kg) body weight to about 1 g / kg body weight, e.g., from about 0.1 μg to about 20 mg / kg body weight, e.g., from about 1 μg to about 5 mg / kg body weight or from about 5 μg to about 1 mg / kg body weight. When used in a combination therapy or composition with another therapeutic agent, the dosage of an active agent of the present invention may optionally be half of the dosages recited above.

[0223] In one embodiment of the present invention, the compound, derivative, or salt of the present invention can be administered to a subject at a dose of 0.5 to 1,333 nmol / kg body weight, for example, 1 to 1,333 nmol / kg body weight, for example, 2 to 1,000 nmol / kg body weight, for example, 4 to 1,333 nmol / kg body weight, for example, 5 to 1,000 nmol / kg body weight, for example, 10 to 750 nmol / kg body weight, for example, 20 to 500 nmol / kg body weight, particularly 30 to 240 nmol / kg body weight. In a preferred embodiment, the highly active compound of the present invention is administered to a subject at a dose of 0.2 to 10 nmol / kg body weight, for example, 0.5 to 5.0 nmol / kg body weight, for example, 1.0 to 2.0 nmol / kg body weight, for example, 1.5 nmol / kg body weight. For a 75 kg subject, such a dose corresponds to a dosage of 37.5 nmol to 100 μmol, e.g., 75 nmol to 100 μmol, e.g., 150 nmol to 100 μmol, e.g., 300 nmol to 100 μmol, e.g., 375 nmol to 75 μmol, e.g., 750 nmol to 56.25 μmol, e.g., 1.5 to 37.5 μmol, especially 2.25 to 18 μmol. In a preferred embodiment of the highly active compounds of the present invention, for a 75 kg subject, such a dose corresponds to a dosage of 15 to 750 nmol, e.g., 37.5 to 375.0 nmol, e.g., 75 to 150 nmol, e.g., 112.5 nmol. The present invention also contemplates dosage ranges bracketed by any of the specific dosage amounts mentioned herein.

[0224] The exact dose will be readily determined by one of ordinary skill in the art based on the potency of the particular compound employed, the route of delivery of the compound, and the age, weight, sex, and physiological condition of the subject.

[0225] For compounds with long blood half-lives, the above-mentioned doses can be administered, for example, once or twice per month, or once, twice, three or four times per week. For preferred compounds, the doses can be administered no more frequently than once per week. Alternatively, for compounds with short blood half-lives, the above-mentioned doses can be administered, for example, once, twice, three or four times per day, or once or twice per week. In some embodiments, the doses can be administered once every 2, 3 or 4 days. According to certain embodiments, the doses can be administered once immediately before each meal. [Example]

[0226] The invention will now be further described with reference to the following non-limiting examples.

[0227] Materials and Methods Peptide synthesis Peptide synthesis was performed on Rink amide MBHA resin. Amino acids were coupled using the Fmoc strategy. Each amino acid was added sequentially from the C-terminus to the N-terminus of the main peptide portion of the molecule. Reagents such as HBTU were used to mediate peptide coupling. The peptide was cleaved from the resin with trifluoroacetic acid in the presence of a scavenger. In the second step, the glutamic acid residue added to the N-terminus of the main peptide portion was replaced and functionalized with an aliphatic dicarboxylic acid group, followed by coupling via one or the other of the carboxylic acid groups.

[0228] Peptides were purified by reverse-phase HPLC. Quality control was performed on all purified peptides, and peptides were shown to be >90% pure by HPLC in two buffer systems in most cases. MALDI-MS showed the expected molecular ion.

[0229] Exemplary Synthesis Using standard Fmoc chemistry, exemplary compound A233 was prepared as follows: 1) Preparation of resin: A solution of Rink amide MBHA resin (0.6 mmol, 1.00 equivalents, Sub 0.3 mmol / g) in DMF was stirred with N2 at 20°C for 2 hours. The mixture was then filtered to obtain the resin. 2) Deprotection: A solution of 20% piperidine in DMF (25.00 mL) was added to the resin, and the mixture was stirred with N for 15 minutes at 20° C. The resin was washed with DMF (5 washes with 25.00 mL) and filtered to obtain the resin. 3) Coupling: A solution of HBTU (648 mg, 1.71 mmol, 2.85 equiv) and Fmoc-Pro-OH (606 mg, 1.8 mmol, 3.00 equiv) in DMF (10.0 mL) was added to the resin along with DIPEA (N,N-diisopropylethylamine) (0.63 m, 3.6 mmol, 6.00 equiv) and stirred with N for 30 min at 20 °C. The resin was then washed five times with DMF (25.0 mL). 4) Steps 3 and 4 were repeated to couple subsequent amino acids. Note:

[0230] [Table 1A]

[0231] [Table 1B]

[0232] Peptide cleavage, disulfide bond formation and purification: 5) The resin was washed twice with MeOH (20.0 mL) and dried under vacuum to yield 8.0 g of peptide resin. Then, 80 mL of cleavage buffer (92.5% TFA / 2.5% Mpr / 2.5% TIS / 2.5% HO) was added to the flask containing the side-chain protected peptide resin at 20 °C, and the mixture was stirred for 2.5 h. The peptide was precipitated with cold tert-butyl methyl ether (800 mL) and centrifuged (3000 rpm for 2 min). The peptide precipitate was washed two more times (400 mL) with tert-butyl methyl ether. The crude peptide was dried, and its identity was confirmed by LCMS. 6) To a solution of the crude peptide (1.32 g) in water (150 mL) and CHCN (100.0 mL) was added a 0.10 M solution of I in MeOH at 20°C dropwise until a yellow color persisted. The mixture was then stirred at 20°C for 2 minutes. After 2 minutes, sodium thiosulfate (0.10 M aqueous solution) was added dropwise until the yellow color disappeared. The mixture was lyophilized to give a crude powder. 7) The residue was purified by preparative HPLC (TFA conditions; 30 °C, A: 0.075% TFA / HO, B: CHCN) to give the example compound (166.9 mg, 40.8 μmol, yield 6.8, purity 92.94%, TFA) as a white solid, the identity of which was confirmed by LCMS.

[0233] Equivalent methods were used for all other peptides described herein.

[0234] Receptor potency of peptides at human amylin receptors. Biological activity was assessed by the ability of peptides to stimulate cAMP production in human embryonic kidney cells using a tetracycline-regulated mammalian expression system (T-Rex-293, Invitrogen) cell line overexpressing human amylin receptor 3 (human calcitonin receptor and receptor activity-modifying protein 3). Cells were transfected with the human calcitonin receptor plasmid and RAMP3 plasmid (Azenta Life Sciences) 24 hours before compound treatment. Individual wells of cells were dosed with a given concentration of test article to test the dose response of each compound. Cellular cAMP was quantified by HTRF (homogeneous time-resolved fluorescence) technology after 30 minutes of peptide stimulation and an additional hour of lysis using a commercially available cAMP kit (Cisbio). Plates were read on a SpectraMax i3x multimode detection platform plate reader, and concentration-response curves were plotted using GraphPad Prism 8.0 (or later). EC values ​​were calculated for each peptide. 50 Values ​​were generated and these were compared to the control for that day.

[0235] In vivo efficacy testing: Single-dose feeding study in male rats Male rats (Charles River, Margate, UK) were used in the animal experiments. Rats were fed ad libitum and housed individually in IVC cages. Animals were randomized into treatment groups stratified by body weight. Peptide solutions were freshly prepared immediately before administration. Control animals received 5% v / v water and 95% NaCl (0.9% w / v), while peptides (either 1.5 or 15 nmol / kg body weight) were resuspended in water for injection. Peptides and vehicle were administered by subcutaneous injection during the early light period (09:00-10:00), and animals were provided with a known amount of food. Animals had free access to food and water throughout the study. Animals were weighed and remaining food was typically measured 24, 48, 72, and 96 hours after dosing, and in some cases 168 hours after dosing. Results presented are for 3 days after dosing.

[0236] Compounds were evaluated for their ability to inhibit food intake and induce weight change, which are reported as two values ​​in Figure 1, referred to as "potency" and "food." "Potency" was scored as two times the difference in weight change (measured in grams) between the treated and vehicle control groups, measured three days after dosing. "Food" was scored by the difference in food intake (measured in grams) consumed by the treated and vehicle control groups, measured three days after dosing.

Claims

1. A compound or derivative of a compound having the formula ABC; or a salt or solvate of the compound or derivative: During the ceremony, A is Q, Q-Ser, Q-Glu or Q-Lys, and Q is a group of formulas: 【Chemistry 1】 and where R is C 8 ~C 28 is an alkylene or alkenylene chain, R 1 Ha-CO 2 H; B is a peptide moiety of 32 or 33 amino acid residues having the sequence XYZ, wherein: X is Lys-Cys, Y is Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6, wherein: Xaa1 is Ser or absent, Xaa2 is Asn, Lys, Ser, or Thr; Xaa3 is Thr or Leu; Xaa4 is Ala or Ser, Xaa5 is Thr, Xaa6 is Cys, Z is Xaa7-Xaa8-Xaa9-Xaa10-Xaa11-Xaa12-Xaa13-Xaa14-Xaa15-Xaa16-Xaa17-Xaa18-Xaa19-Xaa2 0-Xaa21-Xaa22-Xaa23-Xaa24-Xaa25-Xaa26-Xaa27-Xaa28-Xaa29-Xaa30-Xaa31 (SEQ ID NO: 6), in the formula: Xaa7 is Ala, Val or Met; Xaa8 is Thr or Leu; Xaa9 is Gln or Gly; Xaa10 is Arg or Lys; Xaa11 is Leu, Xaa12 is Ala or Ser, Xaa13 is any amino acid, Xaa14 is Glu, Asp or Phe; Xaa15 is Leu, Xaa16 is His, Xaa17 is Lys or Arg; Xaa18 is Leu, Xaa19 is Gln or Lys; Xaa20 is Thr, Xaa21 is Tyr or Phe; Xaa22 is Pro, Xaa23 is Arg or Lys; Xaa24 is Thr, Xaa25 is Gln, Asp, Pro, Lys, or Asn; Xaa26 is Val or Thr; Xaa27 is Gly, Xaa28 is Ser or Ala, Xaa29 is Lys, Asn, Gly or Asp; Xaa30 is Thr or Ala; Xaa31 is any amino acid, C is a terminal -NH bonded to the C-terminus of peptide moiety B 2 It is the base.

2. 2. The compound, derivative or salt of claim 1, wherein a disulfide bridge is present between the side chains of two Cys residues.

3. 3. The compound, derivative or salt of claim 1 or claim 2, wherein Xaa1 is absent.

4. 4. The compound, derivative or salt of claim 3, wherein Xaa2 is Asn, Lys or Ser, Xaa3 is Thr and Xaa4 is Ala.

5. 5. The compound, derivative or salt according to claim 3 or 4, wherein Xaa7 is Ala, Xaa8 is Thr, Xaa9 is Gln, Xaa10 is Arg and Xaa12 is Ala.

6. 6. A compound, derivative or salt according to any one of claims 3 to 5, wherein Xaa13 is Glu, Asp, Gln, Asn or Lys, Xaa14 is Glu, Xaa15 is Leu, Xaa17 is Lys, His or Arg, Xaa18 is Leu, Xaa19 is Gln, Xaa22 is Pro and Xaa24 is Thr.

7. 7. The compound, derivative or salt of claim 6, wherein Xaa13 is Asp, Glu or Gln.

8. 8. A compound, derivative or salt according to claim 6 or claim 7, wherein Xaa17 is Lys.

9. 9. The compound, derivative or salt according to any one of claims 3 to 8, wherein Xaa25 is Pro, Xaa26 is Val, Xaa27 is Gly and Xaa28 is Ser.

10. 9. The compound, derivative or salt according to any one of claims 3 to 8, wherein Xaa29 is Gly and Xaa30 is Thr or Ala.

11. 9. A compound, derivative or salt according to any one of claims 3 to 8, wherein Xaa29 is Asn and Xaa30 is Thr or Ala.

12. 12. The compound, derivative or salt according to claim 10 or 11, wherein Xaa30 is Thr and Xaa31 is Pro.

13. 2. The compound, derivative or salt of claim 1, having an amino acid sequence corresponding to any one of the amino acid sequences listed in the table of FIG.

14. 10. The compound, derivative or salt of claim 1 having the formula ABC: During the ceremony, A is Q-Lys, where Q is a group of formulas: 【Chemistry 2】 and; B is a peptide moiety of 32 amino acid residues having the sequence Lys-Cys-Ser-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Glu-Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Pro-Val-Gly-Ser-Asn-Thr-Pro; C is a terminal -NH bonded to the C-terminus of peptide moiety B 2 It is the base.

15. 15. The compound, derivative or salt of any one of claims 1 to 14, wherein R has an even number of carbon atoms.

16. R is C 16 or C 18 16. The compound, derivative or salt of claim 15, which is an alkylene or alkenylene group.

17. R is C 18 17. The compound, derivative or salt of claim 16, which is a straight-chain alkylene group.

18. 18. A derivative of a compound according to any one of claims 1 to 17, or a salt or solvate of such a derivative, comprising one or more derivatizations selected from amidation, glycosylation, carbamylation, acylation, sulfation, phosphorylation, cyclization, lipidation, pegylation and fusion to another peptide or protein to form a fusion protein.

19. 19. A compound, derivative, salt or solvate according to any one of claims 1 to 18 together with a further therapeutic agent for simultaneous, sequential or separate administration.

20. 20. The compound, derivative, salt or solvate of claim 19, wherein the further therapeutic agent is a GLP-1 receptor agonist, a Y2 (PYY) receptor agonist, a SIP receptor agonist or a glucagon receptor agonist.

21. 20. The compound, derivative, salt or solvate of claim 19, wherein the further therapeutic agent is insulin or an insulin derivative or agonist, GLP-1 or a GLP-1 derivative or agonist, GIP or a GIP derivative or agonist, or glucagon or a glucagon derivative or agonist.

22. 22. A composition comprising a compound, derivative, salt or solvate according to any one of claims 1 to 21 together with a pharmaceutically acceptable carrier.

23. 23. The composition of claim 22, present in a syringe or other administration device for subcutaneous administration to a human.

24. 24. A compound, derivative, salt or solvate according to any one of claims 1 to 21 or a composition according to claim 22 or claim 23 for use as a medicament.

25. 24. A method for treating or preventing a disease or disorder or other unwanted physiological condition in a subject, comprising administering a therapeutically effective amount of a compound, derivative, salt or solvate of any one of claims 1 to 21, or a composition of claim 22 or claim 23.

26. 24. A compound, derivative, salt, or solvate according to any one of claims 1 to 21, or a composition according to claim 22 or claim 23, for use in the prevention or treatment of diabetes, obesity, heart disease, stroke, and non-alcoholic fatty liver disease, improving insulin release in a subject, improving carbohydrate metabolism in a subject, improving the lipid profile in a subject, reducing appetite, reducing food intake, reducing calorie intake, and / or improving carbohydrate tolerance in a subject.

27. 24. A method for treating or preventing diabetes, obesity, heart disease, stroke or non-alcoholic fatty liver disease in a subject, improving insulin release in a subject, improving carbohydrate metabolism in a subject, improving the lipid profile of a subject, improving carbohydrate tolerance in a subject, reducing appetite and / or reducing food intake, reducing calorie intake, said method comprising administering a therapeutically effective amount of a compound, derivative, salt or solvate of any one of claims 1 to 21 or a composition of claim 22 or claim 23.

28. 22. Use of a compound, derivative, salt or solvate according to any one of claims 1 to 21 for the manufacture of a medicament for the prevention or treatment of diabetes, obesity, heart disease, stroke and non-alcoholic fatty liver disease, improving insulin release in a subject, improving carbohydrate metabolism in a subject, improving the lipid profile in a subject, improving carbohydrate tolerance in a subject, reducing appetite, reducing food intake, and / or reducing calorie intake.

29. 24. A method for causing weight loss or preventing weight gain in a subject for cosmetic purposes, comprising administering an effective amount of a compound, derivative, salt or solvate of any one of claims 1 to 21 or a composition of claim 22 or claim 23.

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