Incretin analogs and uses thereof

Abstract

To provide a therapeutic agent available for use in type 2 diabetes mellitus, which enables effective glucose control with weight loss benefits and a favorable side effect profile, and which has sufficiently extended duration of action to allow for dosing at low frequency such as once a day, thrice-weekly, twice-weekly, or once a week.SOLUTION: Incretin analogs are provided that have structural features resulting in balanced activity and extended duration of action at each of glucose-dependent insulinotropic polypeptide (GIP), glucagon-like peptide-1 (GLP-1), and glucagon receptors. Methods also are provided for treating diseases such as diabetes, dyslipidemia, fatty liver disease, metabolic syndrome, non-alcoholic steatohepatitis, and obesity.SELECTED DRAWING: None

Description

Technical Field

[0001] The present disclosure relates to incretin analogs having activity at each of glucose-dependent insulinotropic polypeptide (GIP), glucagon-like peptide-1 (GLP-1), and the glucagon receptor. The incretin analogs described herein provide balanced activity at each of these receptors and have structural features that result in a long duration of action. Such incretin analogs may be useful for treating disorders such as type 2 diabetes (T2DM), dyslipidemia, metabolic syndrome, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (N ASH), and / or obesity. ASH), and / or obesity.

Background Art

[0002] Over the past few decades, the prevalence of diabetes has been continuously increasing. T2DM is the most common form of diabetes, accounting for approximately 9 0% of all diabetes. T2DM is characterized by elevated blood glucose levels caused by insulin resistance. Current standard treatments for T2DM include dietary restriction and exercise, as well as treatment with oral medications and injectable hypoglycemic agents including incretin -based therapies such as GLP-1 receptor agonists.

[0003] GLP-1 is a 36-amino acid peptide, and its major bioactive fragment is a 30-amino acid, C-terminally amidated peptide (GLP-1 that stimulates glucose-dependent insulin secretion and prevents hyperglycemia in diabetic patients. 7-36 Currently, various GLP-1s, including dulaglutide, exenatide, and liraglutide, Analogs are available for treating T2DM. However, many of the currently marketed GLP-1 receptor agonists have their dosages limited by gastrointestinal side effects such as nausea and vomiting. Insulin is considered when treatment with oral medications and incretin-based therapies is inadequate. Despite the available treatment options, a significant number of individuals receiving approved therapies are not achieving glycemic control goals (see, e.g., Casagrande

[0004] et al. (2013) Diabetes Care 36:2271-2279). Uncontrolled diabetes can cause one or more conditions that affect the incidence and mortality of such individuals. One of the major risk factors for T2DM is obesity, and the majority (about 90%) of individuals with T2DM are overweight or obese. Reduction of body fat accumulation has been demonstrated to result in improvement of obesity-related comorbidities including

[0005] hyperglycemia and cardiovascular events. Therefore, there is a need for therapies that are effective in glucose control and weight loss for better disease management. From that perspective, new therapies under investigation include compounds that have activity not only at the GLP-1 receptor but also at one or more other receptors such as the GIP and / or glucagon receptors. In fact, certain No. 4 is an exendin-4 analog that is itself a GLP-1 analog and has triple agonist activity. Analogs of exendin-4 will be described. Similarly, International Patent Application No. WO2014 / 04961 No. 0 and WO2017 / 116204 each describe various analogs having triple agonist activity. In addition, International Patent Application No. WO2017 / 153375 describes glucagon and GLP-1 co-agonists that are also said to have GIP activity.

[0006] Nevertheless, there remains a need for treatments, particularly for T2DM, that can provide effective glucose control with the benefits of weight loss and a favorable side effect profile. In addition, there is a need for therapeutic agents that can be used in a manner with a sufficiently long duration of action to allow for low-frequency administration, such as once daily, three times a week, twice a week, or once a week. SUMMARY OF THE INVENTION

[0007] The incretin analogs described herein are aimed at meeting the above needs. Accordingly, the present disclosure describes incretin analogs that are active at each of the GIP, GLP-1, and glucagon receptors. Advantageously, the incretin analogs described herein have a balanced activity that allows for administration of a dose that provides sufficient activity at each receptor to provide the benefits of agonism at that receptor while avoiding undesirable side effects associated with excessive activity. In addition, the incretin analogs described herein have a long duration of action at each of the GIP, GLP-1, and glucagon receptors, allowing for administration at low frequencies, such as once daily, three times a week, twice a week, or once a week. , Incretin analogs enhance glucose control, cause weight loss and / or improve body composition and have other metabolic benefits, such as reducing proprotein convertase subtilisin / kexin type 9 (PCSK9) and having lipid benefits, and / or increasing bone mass or bone formation or decreasing bone resorption and providing other benefits. The present disclosure also describes effective treatments for other disorders or conditions, including obesity, NAFLD, NASH, dyslipidemia, and / or metabolic disorders. .

[0008] In one embodiment, there is provided an incretin analog comprising the following formula: YX2QGTFTSDYSIX 13 LDKX 17 AX 19 X 20 AFIEYLLX 28 X 29 GPSSX 34 APPPS, wherein X2 is Aib, X 13 is L or α MeL, X 17 is any amino acid having a functional group available for conjugation, the functional group is conjugated to a C 16 -C 22 fatty acid, X 19 is Q or A, X 20 is Ai b, αMeK, Q, or H, X 28 is E or A, X 29 is G or Aib, X 34 is G or Aib (SEQ ID NO: 5), and the C-terminal amino acid is optionally amidated, or a pharmaceutically acceptable salt thereof. is provided. .

[0009] In another embodiment, dyslipidemia, fatty liver disease, metabolic syndrome, NASH , Methods for treating diseases such as obesity and T2DM are provided. Such methods can include at least the step of administering to an individual in need thereof an effective amount of an incretin analog described herein. In some examples, the disease is non-alcoholic fatty liver disease , obesity, NASH, or T2DM.

[0010] In another embodiment, an incretin analog described herein is provided for use in therapy. For example, an incretin analog described herein is for use in the treatment of diseases such as dyslipidemia , non-alcoholic fatty liver disease, metabolic syndrome, NASH, obesity, and T2DM. In some examples, the disease is non-alcoholic fatty liver disease, obesity , NASH, or T2DM.

[0011] In another embodiment, an incretin analog described herein is provided for use in the manufacture of a medicament for treating dyslipidemia, non-alcoholic fatty liver disease , metabolic syndrome, NASH, obesity, and T2DM. In some examples, the disease is non-alcoholic fatty liver disease , obesity, NASH, or T2DM.

[0012] In another embodiment, a pharmaceutical composition is provided comprising an incretin analog described herein and a pharmaceutically acceptable carrier, diluent, or excipient.

[0013] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Any methods and materials similar or equivalent to those described herein can be used with the incretin analog , which can be used in the practice or testing of pharmaceutical compositions and methods, but the preferred methods and materials are described herein.

[0014] Also, references to an element by the indefinite article "a" or "an" do not exclude the possibility that more than one element exists, unless the context clearly requires that there be one and only one element. Thus, the indefinite article "a" or "an" typically means "at least one".

[0015] GIP is a 42 - amino acid peptide (SEQ ID NO: 4), an incretin, that plays a physiological role in glucose homeostasis by stimulating insulin secretion from pancreatic beta - cells in the presence of glucose.

[0016] GLP - 1 is a 36 - amino acid peptide (SEQ ID NO: 2), also an incretin, that has been shown to stimulate glucose - dependent insulin secretion and prevent hyperglycemia in diabetic patients.

[0017] Glucagon is a 29 - amino acid peptide (SEQ ID NO: 1) that binds to and activates the glucagon receptor on hepatocytes to help maintain blood glucose levels and causes the liver to release glucose stored in the form of glycogen through a process called glycogenolysis.

[0018] Oxyntomodulin (OXM) is a 37 - amino acid peptide that contains not only the 29 - amino acid sequence of glucagon, but also an octapeptide carboxy - terminal extension (SEQ ID NO: 3) that activates both the glucagon and GLP - 1 receptors, and has slightly higher potency for the glucagon receptor than for the GLP - 1 receptor. ​​​​

[0019] In addition to T2DM, incretins and analogs thereof having activity in one or more of GIP, GLP-1, and / or glucagon receptors have been described as having potential therapeutic value in a number of other conditions, diseases, or disorders, such as obesity, NAFLD and NASH, dyslipidemia, metabolic syndrome, bone-related disorders, Alzheimer's disease , and Parkinson's disease. For example, see Jall et al. (2017) Mol. Metab. 6:440-446; Carbone et al. (2016) J. Gastroenterol. Hepatol. 31:23-31; Finan et al. (2016) Trends Mol. Med. 22:359-376; Choi et al. (2017) Potent body weight loss and efficacy in a NASH animal model by a nov el long-acting GLP-1 / Glucagon / GIP triple -agonist (HM15211), ADA Poster 1139-P; Din g(2008) J. Bone Miner. Res. 23:536-543; Tai e t al. (2018) Brain Res. 1678:64-74; Muller e t al. (2017) Physiol. Rev. 97:721-766; Finan et al. (2013) Sci. Transl. Med. 5:209; Holsche r (2014) Biochem. Soc. Trans. 42:593-600. See also for reference.

[0020] As used herein, "about" means, for example, within a statistically significant range of a value(s) such as a recited concentration, length, molecular weight, p H, percent sequence identity, time frame, temperature, or volume. Such a value or range can be within a magnitude typically within 20% of a given value or range, more typically within 10% of a given value or range, and even more typically within 5% of a given value or range. The allowable variation encompassed by "about" depends on the particular system under study and can be readily understood by one of ordinary skill in the art. When used herein, with respect to one or more of GIP, GLP-1, or the glucagon receptor, "active," "activate," "activation," etc. mean the ability of a compound, such as an incretin analog described herein, to bind to a receptor(s) and induce a response at the receptor(s), as measured using assays known in the art, such as the in vitro assays described below. When used herein, "an amino acid having a functional group available for conjugation" means any natural or non-natural amino acid having a functional group that can be conjugated to a fatty acid by a linker. Examples of such functional groups include, but are not limited to, alkynyl, alkenyl, amino, azide, bromo, carboxyl, chloro, iodo, and thiol groups. Examples of natural amino acids containing such functional groups include K (amino), C (thiol), E (carboxyl), and D (carboxyl).

[0021] As used herein, "C -C fatty acid" means a fatty acid having 16 to 22 carbon atoms When used herein, with respect to one or more of GIP, GLP-1, or the glucagon receptor, "active," "activate," "activation," etc. mean the ability of a compound, such as an incretin analog described herein, to bind to a receptor(s) and induce a response at the receptor(s), as measured using assays known in the art, such as the in vitro assays described below. When used herein, "an amino acid having a functional group available for conjugation" means any natural or non-natural amino acid having a functional group that can be conjugated to a fatty acid by a linker. Examples of such functional groups include, but are not limited to, alkynyl, alkenyl, amino, azide, bromo, carboxyl, chloro, iodo, and thiol groups. Examples of natural amino acids containing such functional groups include K (amino), C (thiol), E (carboxyl), and D (carboxyl).

[0022] As used herein, "C -C fatty acid" means a fatty acid having 16 to 22 carbon atoms When used herein, "an amino acid having a functional group available for conjugation" means any natural or non-natural amino acid having a functional group that can be conjugated to a fatty acid by a linker. Examples of such functional groups include, but are not limited to, alkynyl, alkenyl, amino, azide, bromo, carboxyl, chloro, iodo, and thiol groups. Examples of natural amino acids containing such functional groups include K (amino), C (thiol), E (carboxyl), and D (carboxyl). As used herein, "C -C

[0023] As used herein, "C 16 -C 22 fatty acid" means a fatty acid having 16 to 22 carbon atoms means a carboxylic acid having. Suitable C for use herein 16 -C 22 fatty acids can be saturated monoacids or saturated diacids. As used herein, "saturated" means that the fatty acid does not contain carbon-carbon double or triple bonds.

[0024] As used herein, "effective amount" means a single or multiple administration to an individual in need thereof, which provides a desired effect in such an individual under diagnosis or treatment, the amount, concentration, or dosage of one or more incretin analogs described herein, or a pharmaceutically acceptable salt thereof, concentration, or dosage. The effective amount can be readily determined by one of ordinary skill in the art by use of known techniques and by observing results obtained in similar circumstances. In determining the effective amount for an individual, a number of factors are considered including, but not limited to, the mammalian species, its size, age, and general health, the particular disease or disorder involved, the degree, involvement, or severity of the disease or disorder, the response of the individual patient, the particular incretin analog administered, the mode of administration, the pharmacokinetic availability characteristics of the preparation administered, the dosage regimen selected, the use of concomitant medications, and other related circumstances. the particular disease or disorder involved, the degree, involvement, or severity of the disease or disorder, the response of the individual patient, the particular incretin analog administered, the mode of administration, the pharmacokinetic availability characteristics of the preparation administered, the dosage regimen selected, the use of concomitant medications, and other related circumstances. the particular disease or disorder involved, the degree, involvement, or severity of the disease or disorder, the response of the individual patient, the particular incretin analog administered, the mode of administration, the pharmacokinetic availability characteristics of the preparation administered, the dosage regimen selected, the use of concomitant medications, and other related circumstances. the particular disease or disorder involved, the degree, involvement, or severity of the disease or disorder, the response of the individual patient, the particular incretin analog administered, the mode of administration, the pharmacokinetic availability characteristics of the preparation administered, the dosage regimen selected, the use of concomitant medications, and other related circumstances. the particular disease or disorder involved, the degree, involvement, or severity of the disease or disorder, the response of the individual patient, the particular incretin analog administered, the mode of administration, the pharmacokinetic availability characteristics of the preparation administered, the dosage regimen selected, the use of concomitant medications, and other related circumstances. the particular disease or disorder involved, the degree, involvement, or severity of the disease or disorder, the response of the individual patient, the particular incretin analog administered, the mode of administration, the pharmacokinetic availability characteristics of the preparation administered, the dosage regimen selected, the use of concomitant medications, and other related circumstances.

[0025] As used herein, "long-acting" means that the binding affinity and activity of the incretin analog persist for a longer period than natural human GIP, GLP-1, and glucagon peptide, continuing for at least once a day, or even less frequently at three times a week, twice a week, or once a week, enabling administration. The time-action profile of the incretin analog can be measured using known pharmacokinetic testing methods such as those utilized in the examples below.

[0026] As used herein, "incretin analog" means a compound that has multiple differences from each of GIP, GLP-1, and glucagon, particularly human GIP (SEQ ID NO: 4), GLP-1 (SEQ ID NO: 2), and glucagon (SEQ ID NO: 1), but has structural similarity. The incretin analogs described herein include amino acid sequences that result in compounds having affinity for the GIP, GLP-1, and glucagon receptors and activity at each of them (i.e., triple agonist activity). and glucagon, particularly human GIP (SEQ ID NO: 4), GLP-1 (SEQ ID NO: 2), and glucagon (SEQ ID NO: 1), but has structural similarity. The incretin analogs described herein include amino acid sequences that result in compounds having affinity for the GIP, GLP-1, and glucagon receptors and activity at each of them (i.e., triple agonist activity). As used herein, "incretin analog" means a compound that has multiple differences from each of GIP, GLP-1, and glucagon, particularly human GIP (SEQ ID NO: 4), GLP-1 (SEQ ID NO: 2), and glucagon (SEQ ID NO: 1), but has structural similarity. The incretin analogs described herein include amino acid sequences that result in compounds having affinity for the GIP, GLP-1, and glucagon receptors and activity at each of them (i.e., triple agonist activity). and glucagon receptors and activity at each of them (i.e., triple agonist activity). )

[0027] As used herein, "an individual in need thereof" means a mammal, such as a human, having a condition, disease, disorder, or symptom that requires treatment or therapy, including those listed herein. As used herein, "an individual in need thereof" means a mammal, such as a human, having a condition, disease, disorder, or symptom that requires treatment or therapy, including those listed herein. As used herein, "an individual in need thereof" means a mammal, such as a human, having a condition, disease, disorder, or symptom that requires treatment or therapy, including those listed herein.

[0028] As used herein, "treat", "treatment", "treating", etc. mean suppressing, delaying, halting, or reversing the progression or severity of an existing condition, disease, disorder, or symptom. As used herein, "treat", "treatment", "treating", etc. mean suppressing, delaying, halting, or reversing the progression or severity of an existing condition, disease, disorder, or symptom. As used herein, "treat", "treatment", "treating", etc. mean suppressing, delaying, halting, or reversing the progression or severity of an existing condition, disease, disorder, or symptom.

[0029] As used herein, with reference to an incretin analog, "triple agonist activity" means an incretin analog having activity at each of the GIP, GLP-1, and glucagon receptors, particularly an analog having balanced and sufficient activity at each receptor to provide the benefits of receptor agonism while avoiding undesirable side effects associated with excessive activity. Also, an incretin analog having triple agonist activity has a long-lasting action at each of the GIP, GLP-1, and glucagon receptors, which is once daily As used herein, with reference to an incretin analog, "triple agonist activity" means an incretin analog having activity at each of the GIP, GLP-1, and glucagon receptors, particularly an analog having balanced and sufficient activity at each receptor to provide the benefits of receptor agonism while avoiding undesirable side effects associated with excessive activity. Also, an incretin analog having triple agonist activity has a long-lasting action at each of the GIP, GLP-1, and glucagon receptors, which is once daily As used herein, with reference to an incretin analog, "triple agonist activity" means an incretin analog having activity at each of the GIP, GLP-1, and glucagon receptors, particularly an analog having balanced and sufficient activity at each receptor to provide the benefits of receptor agonism while avoiding undesirable side effects associated with excessive activity. Also, an incretin analog having triple agonist activity has a long-lasting action at each of the GIP, GLP-1, and glucagon receptors, which is once daily As used herein, with reference to an incretin analog, "triple agonist activity" means an incretin analog having activity at each of the GIP, GLP-1, and glucagon receptors, particularly an analog having balanced and sufficient activity at each receptor to provide the benefits of receptor agonism while avoiding undesirable side effects associated with excessive activity. Also, an incretin analog having triple agonist activity has a long-lasting action at each of the GIP, GLP-1, and glucagon receptors, which is once daily As used herein, with reference to an incretin analog, "triple agonist activity" means an incretin analog having activity at each of the GIP, GLP-1, and glucagon receptors, particularly an analog having balanced and sufficient activity at each receptor to provide the benefits of receptor agonism while avoiding undesirable side effects associated with excessive activity. Also, an incretin analog having triple agonist activity has a long-lasting action at each of the GIP, GLP-1, and glucagon receptors, which is once daily As used herein, with reference to an incretin analog, "triple agonist activity" means an incretin analog having activity at each of the GIP, GLP-1, and glucagon receptors, particularly an analog having balanced and sufficient activity at each receptor to provide the benefits of receptor agonism while avoiding undesirable side effects associated with excessive activity. Also, an incretin analog having triple agonist activity has a long-lasting action at each of the GIP, GLP-1, and glucagon receptors, which is once daily Advantageously enables administration at low frequencies of once, twice, or three times per week.

[0030] The structural features of the incretin analogs described herein provide favorable effects on activity at each receptor, such that they have sufficient activity (i.e., triple agonist activity) at each of the GIP, GLP-1, and glucagon receptors to obtain desirable effects, but do not overwhelm activity at the other two receptors or result in undesirable side effects when administered at a dose sufficient to provide activity at all three receptors. Thus, analogs are provided that have little or no activity at any one receptor. Non-limiting examples of such structural features in certain embodiments include, with reference to SEQ ID NO: 5, L or αMeL at position 13, which has been found to contribute to optimal glucagon and GIP activity, Aib at position 20, which has been found to contribute to optimal GIP activity, acylation at position 17, which has been found to contribute to optimal glucagon activity, and Y at position 25, which has been found to contribute to optimal glucagon and / or GIP activity. Other examples of such structural features include the amino acids described herein at positions 22, 24, and 28-39, which have been found to contribute to optimal binding and potency at all three receptors. The structural features of the incretin analogs described herein also result in analogs having many other beneficial attributes relating to their potential for development as therapeutic agents, including improved solubility of the analogs in aqueous solutions, improved chemical and physical formulation stability, extended pharmacokinetic profiles, and minimized

[0031] potential for immunogenicity. Specific structures that provide such attributes include, but are not limited to, those described herein. The structural features of the incretin analogs described herein also result in analogs having many other beneficial attributes relating to their potential for development as therapeutic agents, including improved solubility of the analogs in aqueous Non-limiting examples of the features include an optimal pharmacokinetic (PK) profile and contributions to developability C 20 acylation at the 17th position with a C fatty acid, an optimal PK profile and contributions to developability Aib, αMeK, Q, or H at the 20th position, which contribute to an optimal PK, immunogenicity , developability, and stability, and amino acids at positions 22, 24, and 28-39 are included.

[0032] The foregoing list of structural features is not comprehensive but illustrative, and the beneficial property combinations of the exemplary analogs described herein are achieved not as a result of any single modification alone, but rather through novel combinations of the structural features described herein and it should be noted. In addition, the above effects of the foregoing list of modifications are not exclusive, as many of these modifications also have other effects important for the properties of the compounds described herein, as described below.

[0033] The amino acid sequences of the incretin analogs described herein typically incorporate naturally occurring amino acids, alpha-methyl substitution residues of natural amino acids (e.g., alpha-methyl leucine (αMeL) and alpha-methyl lysine (αMeK)), and certain other non-natural amino acids, such as alpha-aminoisobutyric acid (Aib), as shown using the standard one-letter code (e.g., L = leucine). The structures of these amino acids are shown below.

Chemical Structure

[0034] As described above, the incretin analogs described herein are not Either has structural similarity but many structural differences. For example, when compared to native human G IP (SEQ ID NO: 4), the incretin analogs described herein have modifications at one or more of positions 2 , 3, 7, 13, 14, 17, 18 - 21, 23 - 25, 28 - 29, and 30 - 42 . In some examples, the incretin analogs described herein have amino acid modifications at each of positions 2 , 3, 7, 13, 14, 17, 18, 20, 21, 23 - 25, 29, and 30 - 42 of native human GIP (SEQ ID NO: 4). In certain examples, the incretin analogs described herein have the following amino acid modifications: Aib at position 2 , Q at position 3, T at position 7, L or αMeL at position 13, L at position 14, optionally through the use of a linker, C -C conjugation to the epsilon-amino group of the K side chain having a fatty acid, a modified K residue at position 17, A at position 18, A at position 21, I at position 23, 16 -C 22 E at position 24, Y at position 25, G or Aib at position 29, and substitution of the amino acids at positions 30 - 42 with an amino acid sequence selected from GPSSGAPPPS (SEQ ID NO: 26) and GPSS - Aib - APPPS (SEQ ID NO: 27) (and truncated analogs). Further in other examples, the incretin analogs described herein also include one or more modifications of A at position 19, α MeK, Aib, or H at position 20, and E at position 28. In certain examples , the incretin analogs described herein are amidated. In addition to the changes described herein, the incretin analogs described herein are capable of binding to and activating each of the GIP, GLP - 1, and glucagon receptors . In certain examples, the incretin analogs described herein are amidated. In addition to the changes described herein, the incretin analogs described herein are capable of binding to and activating each of the GIP, GLP - 1, and glucagon receptors such that the analog can bind to and activate each of the GIP, GLP - 1, and glucagon receptors may include one or more additional amino acid modifications, provided that there is up to...

[0035] As described above, the incretin analogs described herein have a functional group available for conjugation to a natural or unnatural amino acid, for example, a fatty acid moiety conjugated by a linker such conjugation is sometimes referred to as acylation. In certain examples, the amino acid having a functional group available for conjugation can be K, C, E, and D. In certain examples, the amino acid having a functional group available for conjugation is K, where the conjugation is to the epsilon-amino group of the K side chain and is thus directed thereto.

[0036] Acylation of the incretin analogs described herein is at position 17 in SEQ ID NO: 5, which was determined to be the optimal location for including this structure. The fatty acid, and in certain embodiments, the linker, acts as an albumin binder and provides the possibility of generating a long-acting compound.

[0037] The incretin analogs described herein utilize a C -C 16 -C 22 fatty acid that is chemically conjugated to the functional group of the amino acid, either directly or via a linker. The length and composition of the fatty acid affect the half-life of the incretin analog, their potency in in vivo animal models, as well as their solubility and stability. Conjugation to a C -C 16 -C 22 saturated fatty monoacid or diacid results in an incretin analog that exhibits desirable half-life, desirable potency in in vivo animal models, as well as desirable solubility and stability characteristics.

[0038] Saturated C for use herein​​​16 -C 22 Examples of fatty acids include palmitic acid (hexadecanoic acid) (C -monoacid), hexadecanedioic acid (C 16 -diacid), margaric acid (heptadecanoic acid) (C -monoacid), heptadecanedioic acid (C 16 -diacid), stearic acid (C 17 -monoacid), octadecanedioic acid (C -diacid), nonadecyl acid (nonadecanoic acid) (C 17 -monoacid), nonadecanedioic acid (C 18 -diacid), arachidic acid (eicosanoic acid) (C 18 -monoacid), eicosanedioic acid (C -diacid), heneicosylic acid (heneicosanoic acid) (C 19 -monoacid), heneicosanedioic acid (C 19 -diacid), behenic acid (docosanoic acid) (C -monoacid), docosanedioic acid (C 20 -diacid), etc., but are not limited thereto. 20 -diacid), behenic acid (docosanoic acid) (C -monoacid), behenic acid (docosanoic acid) (C 21 -monoacid), behenic acid (docosanoic acid) (C 21 -diacid), docosanoic acid (C -monoacid), docosanedioic acid (C 22 -diacid), etc., but are not limited thereto. 22 -diacid), etc., but are not limited thereto. Not limited to these.

[0039] In certain examples, the C 16 -C 22 fatty acid can be a saturated C 18 -monoacid, a saturated C 18 -diacid, a saturated C1 9-monoacid, a saturated C 19 -diacid, a saturated C 20 -monoacid, a saturated C 20 -diacid, and their branched and substituted derivatives. In more specific examples, the C 16 -C 22 fatty acid can be stearic acid, arachidic acid, and eicosanedioic acid, particularly arachidic acid.

[0040] In some examples, the linker is 1 to 4 amino acids, aminopolyethylene glycol It can have carboxylates, or mixtures thereof. In certain examples, amino Polyethylene glycol carboxylate has the following structure, [Chemical formula] wherein m is any integer from 1 to 12, n is any integer from 1 to 12, and p is 1 or 2.

[0041] In certain examples, the linker can optionally have one or more (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) moieties combined with 1 to 4 amino acids. be possible.

[0042] In examples where the linker contains at least one amino acid, the amino acid can be 1 to 4 Glu or γGlu amino acid residues. In some examples, the linker can contain one or two Glu or γGlu amino acid residues, including their D-form. For example, the linker can contain one or two γGlu amino acid residues either way. Or, the linker can contain 1 to 4 amino acid residues (such as Glu or γGlu amino acids, etc.) used in combination with up to 36 (2-[2-(2-amino-ethoxy)-ethoxy]- acetyl) moieties. Specifically, the linker can be a combination of 1 to 4 Glu or γGlu amino acids and 1 to 4 (2-[2-(2-amino-ethoxy)-ethoxy]- acetyl) moieties. In other examples, the linker can be a combination of 1 or 2 γGlu amino acids and 1 or 2 (2-[2-(2-amino-ethoxy)-ethoxy]- acetyl) moieties. Glu or γGlu amino acids and 1 to 4 (2-[2-(2-amino-ethoxy)-ethoxy]- acetyl) moieties. In other examples, the linker can be a combination of 1 or 2 γGlu amino acids and 1 or 2 (2-[2-(2-amino-ethoxy)-ethoxy]- acetyl) moieties.

[0043] In certain examples, the incretin analogs described herein have a structure of the following formula and have a linker and a fatty acid moiety, (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl)a-(γGlu)b -CO-(CH2)c-CO2H, wherein a is 0, 1, or 2, b is 1 or 2, and c is 16 or 18 is. In certain examples, a is 2, b is 1, c is 18, and its structure is shown below. [Chemical formula] .

[0044] In another specific example, a is 1, b is 2, c is 18, and its structure is shown below. [Chemical formula]

[0045] In another specific example, a is 0, b is 2, c is 18, and its structure is shown below. [Chemical formula]

[0046] In another specific example, a is 1, b is 1, c is 18, and its structure is shown below. [Chemical formula]

[0047] As shown in the chemical structures of Examples 1-20 below, the above linker-fatty acid moiety can be linked to the epsilon (ε)-amino group of the lysine (K) side chain. ​

[0048] For each of GIP, GLP-1, and the glucagon receptor, as described herein The affinity of the incretin analogs can be measured using techniques known in the art for measuring receptor binding, including, for example, those described in the following examples, and is generally expressed as an inhibition constant (Ki) value. The activity of the incretin analogs described herein at each of the receptors can also be measured using techniques known in the art, including, for example, the in vitro activity assays described below, and is generally expressed as an effective concentration 50 (EC 50) value which is the concentration of the compound that causes a maximal half-maximal stimulation in a dose-response curve. 50 50 ) value and is the concentration of the compound that causes a maximal half-maximal stimulation in a dose-response curve.

[0049] The incretin analogs described herein can be formulated as pharmaceutical compositions and administered by parenteral routes (e.g., subcutaneous, intravenous, intraperitoneal, intramuscular, or transdermal). Such pharmaceutical compositions and techniques for preparing them are well known in the art. See, for example, Remington: The Science and Practice of Pharmacy (Troy, Ed., 21 st Edition, Lippincott, Williams & Wilkins, 2006). In certain instances, the incretin analogs are administered subcutaneously. Practice of Pharmacy (Troy, Ed., 21 st nd Edition ), Lippincott, Williams & Wilkins, 2006). In certain instances, the incretin analogs are administered subcutaneously. See also Remington: The Science and Practice of Pharmacy (Troy, Ed., 21st Edition, Lippincott, Williams & Wilkins, 2006). In certain instances, the incretin analogs are administered subcutaneously.

[0050] The incretin analogs described herein can react with any of several inorganic and organic acids / bases to form pharmaceutically acceptable acid / base addition salts. Pharmaceutically acceptable salts and general techniques for preparing them are well known in the art (e.g., ​​​ , Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection and Use, 2 nd Revised Edition (Wiley-VCH, 2011) (see). Pharmaceutically acceptable salts for use herein include sodium salts, trifluoroacetic salts, hydrochloride salts, and / or acetate salts.

[0051] The present disclosure also provides, and thus encompasses, novel intermediates and methods for synthesizing the incretin analogs described herein, or pharmaceutically acceptable salts thereof. The intermediates and incretin analogs described herein can be prepared by various techniques known in the art. For example, methods using chemical synthesis are illustrated in the following examples. The specific synthetic steps for each of the described routes can be combined in different ways to prepare the incretin analogs described herein. Reagents and starting materials are readily available to those skilled in the art.

[0052] The specific incretin analogs described herein are generally effective over a wide dosage range. For example, the dosage for once-weekly administration is within the range of about 0.01 to about 30 mg / person / week, within the range of about 0.1 to about 10 mg / person / week, or even within the range of about 0.1 to about 3 mg / person / week. Thus, the incretin analogs described herein can be administered once daily, three times a week, twice a week, or once a week, particularly once a week.

[0053] The incretin analogs described herein are useful for treating various conditions, disorders, diseases, or symptoms ​It can be used for treatment. In particular, a method for treating T2DM in an individual is provided and such a method comprises at least the step of administering to an individual in need of such treatment an effective amount of an incretin analogue described herein, or a pharmaceutically acceptable salt thereof.

[0054] In addition, a method for treating obesity in an individual is provided, and such a method comprises at least the step of administering to an individual in need of such treatment an effective amount of an incretin analogue described herein, or a pharmaceutically acceptable salt thereof.

[0055] In addition, a method for inducing non-therapeutic weight loss in an individual is provided, and such a method comprises at least the step of administering to an individual in need of such treatment an effective amount of an incretin analogue described herein, or a pharmaceutically acceptable salt thereof.

[0056] In addition, a method for treating metabolic syndrome in an individual is provided, and such a method comprises at least the step of administering to an individual in need of such treatment an effective amount of an incretin analogue described herein, or a pharmaceutically acceptable salt thereof.

[0057] In addition, a method for treating NASH in an individual is provided, and such a method comprises at least the step of administering to an individual in need of such treatment an effective amount of an incretin analogue described herein, or a pharmaceutically acceptable salt thereof.

[0058] In addition, a method for treating NAFLD in an individual is provided, and such a method , administering to an individual in need thereof an effective amount of an incretin as described herein a mimetic, or a pharmaceutically acceptable salt thereof, at least comprising the step of

[0059] In these methods, the effectiveness of the incretin mimetic can be evaluated, for example, by observing a significant decrease in blood glucose, observing a significant increase in insulin, observing a significant decrease in HbA1c, and / or observing a significant decrease in body weight can be.

[0060] Alternatively, the incretin mimetic or a pharmaceutically acceptable salt thereof described herein can be used to improve bone strength in an individual in need thereof. In some examples the individual in need thereof has hypoossification or hypoosteoidosis, or has healed from a fracture, arthromyodynia rehabilitation, orthosis implant, dental implant, and / or spinal fixation. The incretin mimetic described herein can also be used to treat other disorders such as Parkinson's disease or Alzheimer's disease .

Example

[0061] Peptide synthesis Example 1: Example 1 is the compound represented by the following description. Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-2-amino no-ethoxy)-ethoxy]-acetyl)2-(γGlu)-CO-(CH2) 18 -C O2H)AQ-αMeK-AFIEYLLA-Aib-GPSSGAPPPS-NH2( SEQ ID NO: 7).

[0062] The following is a depiction of the structure of Example 1, excluding the residues Aib2, αMeL13, K17, αMeK20, and Aib29, and using the standard one-letter amino acid code, with the structures of these amino acid residues being extended.

Chemical Structure

[0063] The peptide backbone of Example 1 is synthesized using fluorenylmethyloxycarbonyl (Fmoc) / tert-butyl (t-Bu) chemistry on a Symphony 12-Channel Multip lex Peptide Synthesizer (Protein Technolo gies, Inc. Tucson, AZ).

[0064] The resin consists of 1%DVB cross-linked polystyrene (Fmoc-Rink-MBHA Low Loading resin, 100 - 200 mesh, EMD Millipore) with a substitution of 0.3 - 0.4 meq / g. Standard side-chain protecting groups are used. Fmoc-Lys (Mtt)-OH is used for the lysine at position 17, and Boc-Tyr(tBu)-OH) is used for the tyrosine at position 1. Prior to each coupling step (7 minutes × 2 times), the Fmoc group is removed using 20% piperidine in DM F. All standard amino acid coupl ings are carried out at a 9-fold molar excess relative to the theoretical peptide loading, using an equimolar ratio of Fmoc amino acid (0.3 mM), diisopropylcarbodiimide (0. 9 mM), and Oxyma (0.9 mM) for 1 hour for primary amines and 3 hours for secondary amines. The exception is the coupling to Cα-methylated amino acids, which are coupled for 3 hours. After the synthesis of the peptide backbone is completed, the resin is thoroughly washed six times with DCM to remove residual DMF. The Mtt protecting group of lysine at position 17 is selectively removed from the peptide resin by treatment with 30% hexafluoroisopropanol (Oakwood Chemicals) in DCM twice (40 minutes treatment × 2 times).

[0065] Subsequent attachment of the fatty acid-linker moiety is achieved by coupling of 2-[2-(2-Fmoc-amino-ethoxy)-ethoxy]-acetic acid (Fmoc-AEEA-OH, ChemPep, Inc.), Fmoc-glutamic acid α-t-butyl ester (Fmoc-Glu-OtBu, Ark Pharm, Inc.), and mono-OtBu-eicosanedioic acid (WuXi AppTec, Shanghai, China). A three-fold excess of reagents (AA:PyAOP:DIPEA = 1:1:1 mol / mol) is used for each coupling for 1 hour.

[0066] Upon completion of the synthesis, the peptide resin is washed with DCM and air-dried completely. The dried resin is treated with 10 mL of cleavage cocktail (trifluoroacetic acid:water:triisopropylsilane, 95:2.5:2.5 v / v) at room temperature for 2 hours. The resin is filtered off and washed twice with 2 mL of neat TFA each. The combined filtrate is treated with 5-fold cold diethyl ether (-20 °C) to precipitate the crude peptide. Subsequently, the peptide / ether suspension is centrifuged at 3500 rpm for 2 minutes to form a solid pellet, the supernatant is decanted, and the solid pellet is triturated twice more with ether and dried in vacuo. The crude peptide is solubilized in 20% acetonitrile / 20% acetic acid / 60% water and purified by preparative HPLC using 100% acetonitrile and 0.1% TFA / water buffer system. Luna 5μm phenyl-hexyl column (21×250 mm, Phenomenex) with a linear gradient of TFA (30 - 50% acetonitrile in 60 minutes) for RP-HPLC purification. The peptides are purified by RP-HPLC. The purity of the peptides is evaluated using analytical RP-HPLC, and the purity criterion is over 95%. The main pool purity of Example 1 is found to be 98.8%. Subsequent lyophilization of the final major product pool yields the lyophilized peptide TFA salt. The molecular weight is determined by LC-MS (obsd: M+4H+ / 4 = 1226.8, Calc M+4H+ / 4 = 1226.9).

[0067] Example 2: Example 2 is the compound represented by the following description. Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-amino- ethoxy)-ethoxy]-acetyl)-(γGlu)-CO-(CH2) 18 -CO2H )AQHAFIEYLLA-Aib-GPSSGAPPPS-NH2 (SEQ ID NO: 6).

[0068] The following is a depiction of the structure of Example 2 using the standard one-letter amino acid code except for residues Aib2, αMeL13, K17, and Aib29, and the structures of these amino acid residues are extended.

Chemical Structure

[0069] Using a process similar to that described above for Example 1, the peptide backbone is synthesized, conjugated to the fatty acid-linker moiety, the purity is examined, and the molecular weight of Example 2 is confirmed.

[0070] Example 3: ​​​​​​​Example 3 is a compound represented by the following description: Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-amino- Ethoxy)-ethoxy]-acetyl)2-(γGlu)-CO-(CH2) 18 -CO2 H) AQ-αMeK-AFIEYLLEGGPSSGAPPPS-NH2 (SEQ ID NO: 8) .

[0071] The following are canonical except for residues Aib2, αMeL13, K17, and αMeK20: FIG. 1 is a representation of the structure of Example 3 using the standard one-letter amino acid code; The structure of the group is expanded. [ka]

[0072] The peptide backbone was synthesized using a process similar to that described above for Example 1. , conjugate to a fatty acid-linker moiety, check for purity, and confirm molecular weight as in Example 3.

[0073] Example 4: Example 4 is a compound represented by the following description: Y-Aib-QGTFTSDYSILLDKK((2-[2-(2-amino-ethoxy) -ethoxy]-acetyl)-(γGlu)-CO-(CH2) 18 -CO2H)AQ-A ib-AFIEYLLA-Aib-GPSSGAPPPS-NH2 (sequence number 9).

[0074] The following are standard 1 sequences, except for residues Aib2, K17, Aib20, and Aib29: 1 is a representation of the structure of Example 4 using the letter amino acid code, with these amino acid residues The structure is expanded. [ka]

[0075] Using a process similar to that described above for Example 1, synthesize the peptide backbone , conjugate it to the fatty acid-linker moiety, examine the purity, and confirm the molecular weight of Example 4.

[0076] Example 5: Example 5 is a compound represented by the following description. Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-amino- ethoxy)-ethoxy]-acetyl)2-(γGlu)-CO-(CH2) 18 -CO2 H)AAQAFIEYLLE-Aib-GPSSGAPPPS-NH2 (SEQ ID NO: 10) .

[0077] The following is a depiction of the structure of Example 5 using the standard one-letter amino acid code, excluding the residues Aib2, αMeL13, K17, and Aib29, and the structures of these amino acid residues are extended.

Chemical Structure

[0078] Using a process similar to that described above for Example 1, synthesize the peptide backbone , conjugate it to the fatty acid-linker moiety, examine the purity, and confirm the molecular weight of Example 5.

[0079] Example 6: Example 6 is a compound represented by the following description. Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-amino- ethoxy)-ethoxy]-acetyl)2-(γGlu)-CO-(CH2) 18 -CO2 ​H) AAQAFIEYLLEGGPSSGAPPPS-NH2 (SEQ ID NO: 11).

[0080] The following are standard one-letter amino acids, except for residues Aib2, αMeL13, and K17: 1 is a representation of the structure of Example 6 using the acid code. The structures of these amino acid residues are shown in the expanded form. It is stretched. [ka]

[0081] The peptide backbone was synthesized using a process similar to that described above for Example 1. , conjugate to a fatty acid-linker moiety, check for purity, and confirm molecular weight as in Example 6.

[0082] Example 7: Example 7 is a compound represented by the following description: Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-amino- Ethoxy)-ethoxy]-acetyl)2-(γGlu)-CO-(CH2) 18 -CO2 H) AQHAFIEYLLEGGPSSGAPPPS-NH2 (SEQ ID NO: 12).

[0083] The following are standard one-letter amino acids, except for residues Aib2, αMeL13, and K17: 1 is a representation of the structure of Example 7 using the acid code. The structures of these amino acid residues are shown in the expanded form. It is stretched. [ka]

[0084] The peptide backbone was synthesized using a process similar to that described above for Example 1. , conjugate to a fatty acid-linker moiety, check for purity, and confirm molecular weight of Example 7.

[0085] Example 8: Example 8 is a compound represented by the following description. Y-Aib-QGTFTSDYSILLDKK((2-[2-(2-amino-ethoxy) -ethoxy]-acetyl)2-(γGlu)-CO-(CH2) 18 -CO2H)AQ- αMeK-AFIEYLLEGGPSSGAPPPS-NH2 (SEQ ID NO: 13).

[0086] The following is a depiction of the structure of Example 8 using the standard one-letter amino acid code, excluding the residues Aib2, K17, and αMeK20, and the structures of these amino acid residues are expanded.

Chemical Structure

[0087] Using a process similar to that described above for Example 1, synthesize the peptide backbone, conjugate it to the fatty acid-linker portion, examine the purity, and confirm the molecular weight of Example 8.

[0088] Example 9: Example 9 is a compound represented by the following description. Y-Aib-QGTFTSDYSI-αMeL-LDKK((γGlu)2-CO-(C H2) 18 -CO2H)AQHAFIEYLLEGGPSSGAPPPS-NH2 (SEQ ID NO: 14).

[0089] The following is a depiction of the structure of Example 9 using the standard one-letter amino acid code, excluding the residues Aib2, αMeL13, and K17, and the structures of these amino acid residues are expanded.

Chemical Structure

[0090] Using a process similar to that described above for Example 1, the peptide backbone is synthesized , conjugated to the fatty acid-linker moiety, the purity is examined, and the molecular weight of Example 9 is confirmed.

[0091] Example 10: Example 10 is the compound represented by the following description. Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-amino- ethoxy)-ethoxy]-acetyl)-(γGlu)-CO-(CH2) 18 -CO2H )AQHAFIEYLLEGGPSSGAPPPS-NH2 (SEQ ID NO: 15).

[0092] The following is a depiction of the structure of Example 10 using the standard one-letter amino acid code, except for residues Aib2, αMeL13, and K17, and the structures of these amino acid residues are extended.

Chemical formula

[0093] Using a process similar to that described above for Example 1, the peptide backbone is synthesized , conjugated to the fatty acid-linker moiety, the purity is examined, and the molecular weight of Example 10 is confirmed.

[0094] Example 11: Example 11 is the compound represented by the following description. Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-amino- ethoxy)-ethoxy]-acetyl)2-(γGlu)-CO-(CH2) 18 -CO2 H) AQ-Aib-AFIEYLLEGGPSSGAPPPS-NH2 (SEQ ID NO: 16) 。

[0095] The following is a description of the structure of Example 11 using the standard one-letter amino acid code, excluding residues Aib2, αMeL13, K17, and Aib20, and the structures of these amino acid residues are extended. 1-letter amino acid code, and the structures of these amino acid residues are extended. The structure of these amino acid residues is extended.

Chemical Structure

[0096] Using a process similar to that described above for Example 1, synthesize the peptide backbone, conjugate it to the fatty acid-linker moiety, examine the purity, and confirm the molecular weight of Example 11. conjugate it to the fatty acid-linker moiety, examine the purity, and confirm the molecular weight of Example 11.

[0097] Example 12: Example 12 is a compound represented by the following description. Y-Aib-QGTFTSDYSI-αMeL-LDKK ((2-[2-(2-amino- ethoxy)-ethoxy]-acetyl)-(γGlu)-CO-(CH2) 18 -CO2H ) AQ-Aib-AFIEYLLEGGPSSGAPPPS-NH2 (SEQ ID NO: 17).

[0098] The following is a description of the structure of Example 12 using the standard one-letter amino acid code, excluding residues Aib2, αMeL13, K17, and Aib20, and the structures of these amino acid residues are extended. 1-letter amino acid code, and the structures of these amino acid residues are extended. The structure of these amino acid residues is extended.

Chemical Structure

[0099] Using a process similar to that described above for Example 1, synthesize the peptide backbone. , conjugate to the fatty acid-linker moiety, examine the purity, and confirm the molecular weight of Example 12.

[0100] Example 13: Example 13 is a compound represented by the following description. Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-amino- ethoxy)-ethoxy]-acetyl)-(γGlu)-CO-(CH2) 18 -CO2H )AQ-Aib-AFIEYLLE-Aib-GPSSGAPPPS-NH2 (SEQ ID NO. 18).

[0101] The following is a depiction of the structure of Example 13 using the standard one-letter amino acid code, excluding the residues Aib2, αMeL13, K17, Aib20, and Aib29, and the structures of these amino acid residues are extended.

Chemical formula

[0102] Using a process similar to that described above for Example 1, synthesize the peptide backbone , conjugate to the fatty acid-linker moiety, examine the purity, and confirm the molecular weight of Example 13.

[0103] Example 14: Example 14 is a compound represented by the following description. Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-amino- ethoxy)-ethoxy]-acetyl)-(γGlu)-CO-(CH2) 18 -CO2H )AQ-Aib-AFIEYLLE-Aib-GPSS-Aib-APPPS-NH2( SEQ ID NO. 19).

[0104] ​The following, except for residues Aib2, αMeL13, K17, Aib20, Aib29, and Aib34, uses the standard one-letter amino acid code in the depiction of the structure of Example 14, and the structures of these amino acid residues are extended.

Chemical Structure

[0105] Using a process similar to that described above for Example 1, synthesize the peptide backbone, conjugate it to the fatty acid-linker moiety, examine the purity, and confirm the molecular weight of Example 14.

[0106] Example 15: Example 15 is the compound represented by the following description. Y-Aib-QGTFTSDYSILLDKK((2-[2-(2-amino-ethoxy) -ethoxy]-acetyl)2-(γGlu)-CO-(CH2) 18 -CO2H)AQ- Aib-AFIEYLLA-Aib-GPSSGAPPPS-NH2 (SEQ ID NO: 20).

[0107] The following, except for residues Aib2, K17, Aib20, and Aib29, uses the standard 1 letter amino acid code in the depiction of the structure of Example 15, and the structures of these amino acid residues are extended.

Chemical Structure

[0108] Using a process similar to that described above for Example 1, synthesize the peptide backbone, conjugate it to the fatty acid-linker moiety, examine the purity, and confirm the molecular weight of Example 15.

[0109] Example 16: Example 16 is a compound represented by the following description. Y-Aib-QGTFTSDYSILLDKK((2-[2-(2-amino-ethoxy) -ethoxy]-acetyl)2-(γGlu)-CO-(CH2) 18 -CO2H)AQ- Aib-AFIEYLLEGGPSSGAPPPS-NH2 (SEQ ID NO: 21).

[0110] The following is a depiction of the structure of Example 16 using standard one-letter amino acid codes, excluding the residues Aib2, K17, and Aib20, and the structures of these amino acid residues are expanded.

Chemical formula

[0111] Using a process similar to that described above for Example 1, synthesize the peptide backbone , conjugate it to the fatty acid-linker moiety, examine the purity, and confirm the molecular weight of Example 16.

[0112] Example 17: Example 17 is a compound represented by the following description. Y-Aib-QGTFTSDYSILLDKK((2-[2-(2-amino-ethoxy) -ethoxy]-acetyl)2-(γGlu)-CO-(CH2) 18 -CO2H)AQ- Aib-AFIEYLLE-Aib-GPSSGAPPPS-NH2 (SEQ ID NO: 22).

[0113] The following is a depiction of the structure of Example 17 using standard one-letter amino acid codes, excluding the residues Aib2, K17, Aib20, and Aib29, and the structures of these amino acid residues are expanded.

Chemical formula

[0114] Using a process similar to that described above for Example 1, the peptide backbone is synthesized , conjugated to the fatty acid-linker moiety, the purity is examined, and the molecular weight of Example 17 is confirmed.

[0115] Example 18: Example 18 is a compound represented by the following description. Y-Aib-QGTFTSDYSILLDKK((2-[2-(2-amino-ethoxy) -ethoxy]-acetyl)-(γGlu)-CO-(CH2) 18 -CO2H)AQ-A ib-AFIEYLLEGGPSSGAPPPS-NH2 (SEQ ID NO: 23).

[0116] The following is a depiction of the structure of Example 18 using the standard one-letter amino acid code, excluding residues Aib2, K17, and Aib20, and the structures of these amino acid residues are expanded .

Chemical Structure

[0117] Using a process similar to that described above for Example 1, the peptide backbone is synthesized , conjugated to the fatty acid-linker moiety, the purity is examined, and the molecular weight of Example 18 is confirmed.

[0118] Example 19: Example 19 is a compound represented by the following description. Y-Aib-QGTFTSDYSILLDKK((2-[2-(2-amino-ethoxy) -ethoxy]-acetyl)-(γGlu)2-CO-(CH2) 18 -CO2H)AQ- Aib - AFIEYLLE - Aib - GPSSGAPPPS - NH2 (SEQ ID NO: 24).

[0119] The following is a depiction of the structure of Example 19 using the standard one - letter amino acid code, excluding the residues Aib2, K17, Aib20, and Aib29, and the structures of these amino acid residues are extended. [Chemical Structure]

[0120] Using a process similar to that described above for Example 1, synthesize the peptide backbone, conjugate it to the fatty acid - linker moiety, examine the purity, and confirm the molecular weight of Example 19.

[0121] Example 20: Example 20 is a compound represented by the following description. Y - Aib - QGTFTSDYSILLDKK ((2 - [2 - (2 - amino - ethoxy) - ethoxy] - acetyl) - (γGlu) - CO - (CH2) 18 - CO2H)AQ - A ib - AFIEYLLE - Aib - GPSSGAPPPS - NH2 (SEQ ID NO: 25).

[0122] The following is a depiction of the structure of Example 20 using the standard one - letter amino acid code, excluding the residues Aib2, K17, Aib20, and Aib29, and the structures of these amino acid residues are extended. [Chemical Structure]

[0123] Using a process similar to that described above for Example 1, synthesize the peptide backbone, conjugate it to the fatty acid - linker moiety, examine the purity, and confirm the molecular weight of Example 20. ​​​​

[0124] In vitro function Binding affinity: Perform a radioligand competition binding assay to determine the equilibrium dissociation constants of exemplary compounds and comparator moieties Such assays are Scintillation Proximity Assay ( SPA) methods and transfected HEK293 cells overexpressing human GIP receptor (GIPR), GLP-1 receptor (GLP-1R), or human glucagon receptor (GcgR) using membranes prepared from the cells. The assay is performed in the presence of bacitracin as a non-specific blocker to prevent the sialylated moiety of the test analog from binding to the protein components (e.g., albumin) used in the standard assay buffer.

[0125] The competition curve is plotted as percent specific inhibition (y-axis) versus the log concentration of the compound (x-axis) and analyzed using a four-parameter non-linear regression fit with variable slope (ABase or Genedata). K values are calculated according to the equation K = IC

[0126] / (1+(D / K )) where IC is the concentration of the compound that results in 50% inhibition of binding, D i is the concentration of the radioligand used in the assay, and K i is the equilibrium dissociation constant of the receptor and radioligand determined from saturation binding analysis (shown in Table 1 below). 50 / (1+(D / K d )) and is calculated according to the equation where IC is the concentration of the compound that results in 50% inhibition of binding, D 50 is the concentration of the radioligand used in the assay, and K is the equilibrium dissociation constant of the receptor and radioligand determined from saturation binding analysis (shown in Table 1 below). d is determined from saturation binding analysis and is the equilibrium dissociation constant of the receptor and radioligand (shown in Table 1 below).

Table 1

Table 2

[0127] As shown in Table 2, exemplary analogs have binding affinity at each of the GIP, GLP-1, and glucagon receptors.

[0128] Functional activity: Functional activity is determined in GIP-R-GLP-1R- and GcgR-expressing HEK-293 cell lines. In a 20 μl assay volume, peptides in DMEM (Gibco catalog number 31053) supplemented with 1X GlutaMAX (Gibco catalog number 35050), 0.25% FBS (fetal bovine serum, Gibco catalog number 26400), 0.05% fraction V BSA (bovine serum albumin, Gibco catalog number 15260), 250 μM IBMX, and 20 mM HEPES (Gibco catalog number 15630) (20 point CRC, 2.75-fold Labcyte Echo direct dilution) are used to treat each receptor overexpressing cell line.

[0129] After incubation for 60 minutes at room temperature, the resulting increase in intracellular cAMP is quantitatively determined using the CisBio cAMP Dynamic 2 HTRF assay kit (62AM4PEJ). Briefly, intracellular cAMP levels are detected by adding the cAMP-d2 conjugate in cell lysis buffer, followed by the antibody, anti-cAMP-cAMP-Eu-cryptate, also in cell lysis buffer. The resulting competitive assay is incubated at room temperature for at least 60 minutes and then the luminescence at 320 nm excitation and 665 nm and 620 nm emission is measured on a PerkinElmer Envision. 3+ ​​​​​​​​​​​​​​​Detect using the Envision unit (luminescence at 665 nm / 620 nm). *10,000) is inversely proportional to the amount of cAMP present and is converted to nM cAMP per well using a cAMP standard curve.

[0130] The amount of cAMP (nM) generated in each well is converted to a percentage of the maximum response observed with either human GLP-1(7-36)NH2, human Gcg, or human GIP(1-42)NH2. The relative EC values are derived by non-linear regression analysis using the maximum response percentage vs. added peptide concentration fit to a four-parameter logistic equation. 50

[0131] Data for exemplary analogs and hGIP(1-42)NH2, hGLP-1(7-36)NH2 , and hGcg are shown in Table 3 below.

Table 3

[0132] As seen in Table 3, in the presence of FBS and BSA, the exemplary analogs have agonist activity lower than that of the native ligands, as determined by the human GIP-R, GLP-1R, and GcgR cAMP cA MP assay.

[0133] An additional set of cAMP assays is performed in HEK293 cells expressing the human GLP-1, GIP, and glucagon receptors. The assays are performed using homogeneous time-resolved fluorescence measurement and in the presence of casein (instead of serum albumin) as a non-specific blocker that does not interact with the fatty acid moiety of the molecules being analyzed. Exemplary analogs and controls are performed in the presence of casein (instead of serum albumin). ​​​​​Determine the intrinsic efficacy of the impaler molecule.

[0134] Intracellular cAMP levels are determined by extrapolation using a standard curve. The dose of the compound Response curves are normalized to the minimum (buffer only) and maximum (maximum concentration of each control ligand) and plotted as a percentage of the stimulated response and fitted to a variable slope 4-parameter nonlinear regression fit (Genedata Screener 13) for analysis. EC 50 is the concentration of the compound that causes a maximum half-maximal simulation in the dose-response curve.

[0135] The data are provided in Table 4 below.

Table 4

[0136] As seen in Table 4, the exemplary analogs stimulate cAMP from human GIP, GLP-1, and glucagon receptors in the presence of 0.1% casein.

[0137] In vivo studies Pharmacokinetics in male Sprague Dawley rats: The pharmacokinetics of the exemplary analogs are evaluated after a single subcutaneous administration of 100 nM / kg to male Sprague Dawley rats. Blood samples are collected over 120 hours and pharmacokinetic parameters are calculated using the resulting individual plasma concentrations. Peptide plasma (K3EDTA) concentrations are determined using a validated LC / MS method that measures the intact mass of the analog. Each peptide and analog as an internal standard is extracted from 100% specific plasma using methanol containing 0.1% formic acid. For LC / MS detection, Thermo Q-Exactive ​​​​​​ve, high-resolution instruments, and Thermo Easy Spray PepMap are combined. The average pharmacokinetic parameters are shown in Table 5. As shown in Table 5, the exemplary analogs exhibit an extended pharmacokinetic profile.

Table 5

[0138] In Vivo Effect on Insulin Secretion in Male Wistar Rats:

[0139] Using an intravenous glucose tolerance test (ivGTT) in rats (male Wistar), the insulin secretory capacity of the exemplary analogs is estimated. The GLP-1 RA semaglutide is used as a positive control. Rats surgically implanted with cannulas in the jugular vein and carotid artery (Envigo, Indianapolis, IN, 280 - 320 grams) are housed one per polycarbonate cage with a filter top. The rats are maintained at 21°C on a 12-hour light / dark cycle and have free access to 2014 Teklad Global Diet (Envigo, Indianapolis) and deionized water. The rats are randomized by body weight and the exemplary analog is administered subcutaneously (sc) at 1.5 mL / kg 16 hours prior to glucose administration, followed by fasting. A typical analog at a stock concentration of 211 nM is diluted in Tris buffer pH 8.0 to 6.667 nMol / mL, 2.0 nMol / mL, 0.667 nMol / mL, 0.2 nMol / mL, and the doses tested are vehicle, 1, 3, and 10 nMol / kg, and in some cases 0.3 and 30 nMol / kg. Semaglutide is used as a positive control, and its ins ​​​​​​​​​​​​​The effect of phosphorus secretion is measured in both implementations of its own test (vehicle and 1, 3, 10, and 30 nMol / kg doses) and the exemplary analog (10 nMol / kg dose). Blood samples at the 0 time point are collected into EDTA tubes and then glucose is administered (0.5

[0140] mg / kg, 5 mL / kg). Blood samples are collected for glucose and insulin levels at 2, 4, 6, 10 mg / kg, 5 mL / kg). Blood samples are collected for glucose and insulin levels at 2, 4, 6, 10, , 20, and 30 minutes after intravenous administration of glucose. Plasma insulin is determined using an electrochemiluminescence assay (Meso Scale, Gaithersbur g, MD). The area under the insulin curve (AUC) is examined and n = 6 animals per group is compared to the vehicle control. Statistical analysis is performed using JMP in one-way analysis of variance followed by Dunnett's comparison to the vehicle control. The data are provided in Table 6 below.

[0141] Statistical analysis is performed using JMP in one-way analysis of variance followed by Dunnett's comparison to the vehicle control. The data are provided in Table 6 below.

Table 6

[0142] As seen in Table 6, the exemplary analog shows a dose-dependent increase in insulin secretion.

[0143] Study in diet-induced obese C57BL / 6 mice: The exemplary incretin analogs described herein are proposed as a treatment for metabolic syndrome, which is a collection of not only diabetes but also insulin resistance and co-existing diseases associated with diabetes (such as dyslipidemia, obesity, hepatic steatosis, etc.). To investigate the effect of the exemplary analogs on parameters such as weight loss, metabolism, body composition, and hepatic steatosis, they was administered to C57BL / 6 diet-induced obesity (DIO) mice. These animals do not have diabetes but exhibit insulin resistance, dyslipidemia, and hepatic steatosis, which are all characteristics of metabolic syndrome after being fed a high-fat diet for 18 weeks.

[0144] Specifically, 24- to 25-week-old DIO male C57BL / 6 mice maintained on a calorie-rich diet are used in the following studies. A temperature-controlled (24 °C ) facility with a 12-hour light / dark cycle (lights on at 22:00) and free access to diet (TD95217) and water is used to house the mice individually. After a minimum of 2 weeks of acclimation to the facility, the animals are randomized according to their body weight so that each experimental group of animals has a similar body weight. The body weight ranges from 40 to 51 g.

[0145] All groups contain 5 to 6 mice. Vehicle, an exemplary analog dissolved in vehicle (40 mM Tris-HCl at pH 8.0), and semaglutide dissolved in vehicle are administered to freely fed DIO mice by subcutaneous (SC) injection (10 mL / kg) 3 days every 15 days, 30 minutes to 90 minutes before the start of the dark cycle. SC injections are performed on days 1, 4, 7, 10, and 13. Body weight and food intake are measured throughout the study. The absolute change in body weight is calculated by subtracting the body weight of the same animal before the first injection of vehicle, analog, or semaglutide. On days 0 and 15, total fat mass is measured by nuclear magnetic resonance (NMR) using an Echo Medical System Instrument (Houston, TX). On day 15, the animals are sacrificed before the dark period

[0146] ​​​​​​ Remove the liver and freeze it. Determine from the liver homogenate collected at slaughter the liver triglycerides and plasma cholesterol using a Hitachi Modular r P clinical analyzer.

[0147] Data are shown as mean ± SEM of 5 - 6 animals per group in Tables 7 and 8 below. Statistical analysis is performed using repeated measures ANOVA followed by Dunnett's method for multiple comparisons. A significant difference is identified at p < 0.05.

Table 7

[0148] As seen in Table 7 above, the exemplary analogs show a dose - dependent decrease in body weight.

[0149] Data for metabolic parameters at a dose of 3 nmol / kg are provided in Table 8 below.

Table 8

[0150] In addition to substantial weight loss, as seen in Table 8, the exemplary analogs reduce blood glucose, plasma insulin (as an indication of increased insulin sensitivity), and plasma cholesterol, and improve liver health as indicated by a decrease in plasma ALT and liver triglycerides.

[0151] SEQ ID NO: 1 - Human Glucagon HSQGTFTSDYSKYLDSRRAQDFVQWLMNT SEQ ID NO: 2 - Human GLP - 1(7 - 36) amide HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR - NH2 SEQ ID NO: 3 - Human OXM HSQGTFTSDYSKYLDSRRAQDFVQWLMNTKRNRNNIA Sequence number 4 - Human GIP YAEGTFISDYSIAMDKIHQQDFVNWLLAQKGKKNDWKHNI TQ Sequence number 5 - Incretin analog YX2QGTFTSDYSIX 13 LDKX 17 AX 19 X 20 AFIEYLLX 28 X 29 GPSSX 34 APPPS In the formula, X2 is Aib, X 13 is L or αMeL, X 17 is any amino acid having a functional group available for conjugation, and the functional group is C1 6 - C 22 conjugated to a fatty acid, X 19 is Q or A, X 20 is Aib, αMeK, Q, or H, X 28 is E or A, X 29 is G or Aib, X 34 is G or Aib Sequence number 6 - Incretin analog Y - Aib - QGTFTSDYSI - αMeL - LDKK((2 - [2 - (2 - amino - ethoxy) - ethoxy] - acetyl) - (γGlu) - CO - (CH2) 18 - CO2H )AQHAFIEYLLA - Aib - GPSSGAPPPS - NH2 Sequence number 7 - Incretin analog Y - Aib - QGTFTSDYSI - αMeL - LDKK((2 - [2 - (2 - amino - (2-(2-(2-Amino-ethoxy)-ethoxy)-acetyl)-2-(γGlu)-CO-(CH2) 18 -CO2 H)AQ-αMeK-AFIEYLLA-Aib-GPSSGAPPPS-NH2 SEQ ID NO: 8 - Incretin analog Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-Amino- (2-(2-(2-Amino-ethoxy)-ethoxy)-acetyl)-2-(γGlu)-CO-(CH2) 18 -CO2 H)AQ-αMeK-AFIEYLLEGGPSSGAPPPS-NH2 SEQ ID NO: 9 - Incretin analog Y-Aib-QGTFTSDYSILLDKK((2-[2-(2-Amino-ethoxy) -ethoxy]-acetyl)-(γGlu)-CO-(CH2) 18 -CO2H)AQ-A ib-AFIEYLLA-Aib-GPSSGAPPPS-NH2 SEQ ID NO: 10 - Incretin analog Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-Amino- (2-(2-(2-Amino-ethoxy)-ethoxy)-acetyl)-2-(γGlu)-CO-(CH2) 18 -CO2 H)AAQAFIEYLLE-Aib-GPSSGAPPPS-NH2 SEQ ID NO: 11 - Incretin analog Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-Amino- (2-(2-(2-Amino-ethoxy)-ethoxy)-acetyl)-2-(γGlu)-CO-(CH2) 18 -CO2 H)AAQAFIEYLLEGGPSSGAPPPS-NH2 SEQ ID NO: 12 - Incretin analog Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-Amino- (2-(2-(2-Amino-ethoxy)-ethoxy)-acetyl)-2-(γGlu)-CO-(CH2) 18-CO2 H)AQHAFIEYLLEGGPSSGAPPPS-NH2 SEQ ID NO:13 - Incretin analog Y-Aib-QGTFTSDYSILLDKK((2-[2-(2-amino-ethoxy) -ethoxy]-acetyl)2-(γGlu)-CO-(CH2) 18 -CO2H)AQ- αMeK-AFIEYLLEGGPSSGAPPPS-NH2 SEQ ID NO:14 - Incretin analog Y-Aib-QGTFTSDYSI-αMeL-LDKK((γGlu)2-CO-(C H2) 18 -CO2H)AQHAFIEYLLEGGPSSGAPPPS-NH2 SEQ ID NO:15 - Incretin analog Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-amino- ethoxy)-ethoxy]-acetyl)-(γGlu)-CO-(CH2) 18 -CO2H )AQHAFIEYLLEGGPSSGAPPPS-NH2 SEQ ID NO:16 - Incretin analog Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-amino- ethoxy)-ethoxy]-acetyl)2-(γGlu)-CO-(CH2) 18 -CO2 H)AQ-Aib-AFIEYLLEGGPSSGAPPPS-NH2 SEQ ID NO:17 - Incretin analog Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-amino- ethoxy)-ethoxy]-acetyl)-(γGlu)-CO-(CH2) 18 -CO2H )AQ-Aib-AFIEYLLEGGPSSGAPPPS-NH2 SEQ ID NO:18 - Incretin analog Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-amino- ethoxy)-ethoxy]-acetyl)-(γGlu)-CO-(CH2) 18 -CO2H )AQ-Aib-AFIEYLLE-Aib-GPSSGAPPPS-NH2 SEQ ID NO:19-incretin analog Y-Aib-QGTFTSDYSI-αMeL-LDKK((2-[2-(2-amino- ethoxy)-ethoxy]-acetyl)-(γGlu)-CO-(CH2) 18 -CO2H )AQ-Aib-AFIEYLLE-Aib-GPSS-Aib-APPPS-NH2 SEQ ID NO:20-incretin analog Y-Aib-QGTFTSDYSILLDKK((2-[2-(2-amino-ethoxy) -ethoxy]-acetyl)2-(γGlu)-CO-(CH2) 18 -CO2H)AQ- Aib-AFIEYLLA-Aib-GPSSGAPPPS-NH2 SEQ ID NO:21-incretin analog Y-Aib-QGTFTSDYSILLDKK((2-[2-(2-amino-ethoxy) -ethoxy]-acetyl)2-(γGlu)-CO-(CH2) 18 -CO2H)AQ- Aib-AFIEYLLEGGPSSGAPPPS-NH2 SEQ ID NO:22-incretin analog Y-Aib-QGTFTSDYSILLDKK((2-[2-(2-amino-ethoxy) -ethoxy]-acetyl)2-(γGlu)-CO-(CH2) 18 -CO2H)AQ- Aib-AFIEYLLE-Aib-GPSSGAPPPS-NH2 SEQ ID NO:23-incretin analog Y-Aib-QGTFTSDYSILLDKK((2-[2-(2-amino-ethoxy) -Ethoxy]-acetyl)-(γGlu)-CO-(CH2) 18 -CO2H)AQ-A ib-AFIEYLLEGGPSSGAPPPS-NH2 SEQ ID NO: 24 - Incretin analog Y-Aib-QGTFTSDYSILLDKK((2-[2-(2-Amino-ethoxy) -Ethoxy]-acetyl)-(γGlu)2-CO-(CH2) 18 -CO2H)AQ- Aib-AFIEYLLE-Aib-GPSSGAPPPS-NH2 SEQ ID NO: 25 - Incretin analog Y-Aib-QGTFTSDYSILLDKK((2-[2-(2-Amino-ethoxy) -Ethoxy]-acetyl)-(γGlu)-CO-(CH2) 18 -CO2H)AQ-A ib-AFIEYLLE-Aib-GPSSGAPPPS-NH2 SEQ ID NO: 26 - Artificial sequence GPSSGAPPPS SEQ ID NO: 27 - Artificial sequence GPSS-Aib-APPPS

Claims

1. 1. An incretin analog comprising: YX ２ QGTFTSDYSIX １３ LDKX １７ AX １９ X ２０ AFIEYLLX ２８ X ２９ GPSSX ３４ APPPS During the ceremony, X ２ is Aib, X １３ is L or αMeL, X １７ is any amino acid having a functional group available for conjugation, said functional group being C １６ -C ２２ Conjugated to fatty acids X １９ is Q or A, X ２０ is Aib, αMeK, Q, or H; X ２８ is E or A, X ２９ is G or Aib, X ３４ is G or Aib, (SEQ ID NO:5) incretin analogs, wherein the C-terminal amino acid is optionally amidated; or a pharma- ceutically acceptable salt thereof.

2. X １７ The amino acids having the functional groups available for conjugation at the positions K, C, E, and 2. The incretin analog of claim 1, wherein the incretin analog is selected from the group consisting of D and D.

3. X １７ The amino acid having the functional group available for conjugation at position A is K.

2. An incretin analog according to claim 1.

4. X １７ the amino acid having the functional group available for conjugation at position C; １６ -C ２２ The fatty acid is conjugated by a linker between the amino acid and the fatty acid.

4. An incretin analog according to any one of 1 to 3.

5. The incretin analog of claim 4, wherein the linker comprises 1 to 4 amino acids.

6. 6. The incretin analog of claim 5, wherein the amino acid is Glu or γGlu. 。

7. The linker is Yellow ２ -CH ２ -[O-CH ２ -CH ２ ] ｍ -O-(CH ２ ) ｐ -CO} ｎ In the formula, m is an integer of 1 to 12, and n is an integer of 1 to 1. 2, and p is 1 or 2. Incretin analogues.

8. The linker is selected from the group consisting of 1 to 4 (2-[2-(2-amino-ethoxy)-ethoxy]-amino The incretin analog of any one of claims 4 to 7, further comprising a cetyl) moiety.

9. X １７ is chemically linked to the epsilon-amino group of the K side chain, which has the structure is K modified by (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) ａ - (γGlu) ｂ -CO-(CH ２ ) ｃ -CO ２ H, where a is 0, 1, or 2, and b is 1 or The incretin analog of claim 1 , wherein n is 2 and c is an integer from 16 to 18.

10. 10. The incretin analog of claim 9, wherein a is 1.

11. 10. The incretin analog of claim 9, wherein a is 2.

12. The incretin analog of any one of claims 9 to 11, wherein b is 1.

13. The incretin analog of any one of claims 9 to 11, wherein b is 2.

14. The incretin analog of any one of claims 9 to 13, wherein c is 18.

15. X １３ The incretin analog according to any one of claims 1 to 14, wherein body.

16. X １３ The incretin analog of any one of claims 1 to 14, wherein is L.

17. X ２０ The incretin analog of any one of claims 1 to 16, wherein is Aib. 。

18. X ２８ The incretin analog of any one of claims 1 to 17, wherein is E.

19. X ２９ The incretin analog of any one of claims 1 to 18, wherein is G.

20. X ３４ The incretin analog of any one of claims 1 to 19, wherein is G.

21. X １９ The incretin analog of any one of claims 1 to 20, wherein is A.

22. a is 1, b is 1, c is 18, and X １３ is αMeL, and X １９ is A and X ２０ is Aib, and X ２８ is E and X ２９ But G , X ３４ The incretin analog of claim 9 , wherein is G.

23. SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, An incretin having a formula selected from the group consisting of SEQ ID NO:21, and SEQ ID NO:

23. Analogs.

24. Diabetes, obesity, fatty liver disease, nonalcoholic steatohepatitis, dyslipidemia, and metabolic syndrome 16. A method for treating a disease selected from the group consisting of rheumatoid arthritis, ... and rheumatoid arthritis, said method comprising the steps of: 、 An effective amount of the incretin according to any one of claims 1 to 23 is administered to an individual in need thereof. The method comprises administering an isolin analog to said subject.

25. 1. A method of treating type II diabetes, the method comprising: An effective amount of the incretin analogue according to any one of claims 1 to 23 is administered to an individual in need thereof. The method comprises administering an analog.

26. An incretin analogue according to any one of claims 1 to 23; A pharmaceutical composition comprising a pharma- ceutically acceptable carrier, diluent, or excipient.

27. Diabetes, dyslipidemia, fatty liver disease, metabolic syndrome, non-alcoholic fat and obesity.

24. The incretin analog of any one of 1-23.

28. A compound according to any one of claims 1 to 23 for use in the treatment of type II diabetes. Ancretin analogue.

29. Diabetes, dyslipidemia, fatty liver disease, metabolic syndrome, non-alcoholic fat and obesity. Use of an incretin analogue according to any one of claims 1 to 23.

30. A compound according to any one of claims 1 to 23 in the manufacture of a medicament for treating type II diabetes. Uses of the incretin analogs described herein.

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