Combination Therapy

Abstract

The present invention relates to therapeutic methods using acylated compounds with dual agonist activity at the GLP-1 (glucagon-like peptide 1) receptor and the GLP-2 (glucagon-like peptide 2) receptor, and peptide hormones. In particular, the present invention relates to the administration of dual GLP-1 / GLP-2 agonist peptides and amylin for the regulation of body weight and for the prevention or treatment of obesity and related conditions.

Description

[Technical Field]

[0001] The present invention relates to therapeutic methods using a combination of acylated compounds with dual agonist activity at the glucagon-like peptide 1 (GLP-1) receptor and the glucagon-like peptide 2 (GLP-2) receptor, and a peptide hormone. In particular, the present invention relates to the combination of a dual GLP-1 / GLP-2 agonist peptide and an amylin analog for the regulation of body weight and for the prevention or treatment of obesity and related conditions. [Background technology]

[0002] Obesity is now a major public health problem in most developed countries and is correlated with the development of several serious conditions, such as cardiovascular disease, type 2 diabetes, sleep apnea, and certain cancers. The standard treatment for obesity is lifestyle intervention, including reduced energy intake and increased physical activity. However, while such interventions can achieve initial success, patients often have difficulty maintaining these lifestyle changes over the long term and making the achieved weight loss permanent.

[0003] GLP-1 is released from the gastrointestinal tract in response to food ingestion, thus acting as a satiety signal and reducing food intake (Madsbad, S., 2014, Diabetes Obes Metab, 16: 9-21). Evidence suggests that the effects of GLP-1 may be reduced in obese subjects, suggesting that GLP-1 agonists may be promising in the treatment of obesity.

[0004] Intestinal tissue is responsible for the production of both human glucagon-like peptide 1 (GLP-1(7-36)) and human glucagon-like peptide 2 (GLP-2(1-33)), which are produced by the same cells. Human GLP-2 has the sequence: Hy-His-Ala-Asp-Gly-Ser-Phe-Ser-Asp-Glu-Met-Asn-Thr-Ile-Leu-Asp-Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn-Trp-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp-OH (SEQ ID NO: 1) It is a peptide consisting of 33 amino acids.

[0005] GLP-2 is derived from specific post-translational processing of proglucagon in enteroendocrine L cells of the intestinal tract and in specific regions of the brainstem. GLP-2 binds to a single G protein-coupled receptor and belongs to the class II glucagon secretin family.

[0006] GLP-2 is co-secreted with GLP-1, oxyntomodulin, and glicentin in response to nutritional intake. Human GLP-1 has the sequence: Hy-His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-Gly-NH2 (SEQ ID NO: 2) It is produced as a peptide consisting of 30 amino acids having the formula:

[0007] GLP-1 has been described as a physiological incretin hormone and has therefore been reported to increase insulin response after oral ingestion of glucose or fat in most cases. However, it is generally known that GLP-1 reduces glucagon levels, has a beneficial effect on inhibiting rapid intestinal motility (Tolessa et al., 1998, Dig. Dis. Sci. 43(10): 2284-90), and delays gastric emptying.

[0008] A major drawback of GLP-1 therapy is that a large proportion of patients taking known GLP-1 agonists experience side effects of nausea and vomiting (Filippatos et al., 2014 / 15, Rev Diabet Stud., 11(3): 202-230). Because of these side effects, it is generally necessary to gradually increase the dose of GLP-1 agonists from a low starting dose to minimize these side effects. Indeed, clinical trial data for the GLP-1 agonist semaglutide show that nausea and vomiting are common among patients upon administration, even when low doses of the drug are initially administered (Wilding et al., 2021, N Engl J Med; 384:989-1002). These side effects are undesirable because they tend to reduce patient compliance with treatment.

[0009] Thus, there remains a need for therapeutic agents with GLP-1 agonist activity that are effective in treating obesity and related conditions while not exhibiting the expected side effects of nausea and vomiting upon administration.

[0010] WO 2013 / 164484 discloses GLP-2 analogues that contain one or more substitutions compared to h[Gly2]GLP-2 and may have modified GLP-1 activity properties, and their medical uses.

[0011] WO 2018 / 104561 discloses peptides with dual GLP-1 and GLP-2 agonist activity and proposes their medical uses, and PCT / EP2022 / 074420 discloses specific dosing regimes for the treatment of obesity and related conditions.

[0012] Amylin is a member of a family of peptide hormones that includes amylin, calcitonin, calcitonin gene-related peptide, adrenomedullin, and intermedin (intermedin is also known as AFP-6), which have been implicated in various metabolic diseases and disorders. Human amylin was first isolated, purified, and characterized as a major component of amyloid deposits in pancreatic islets of type 2 diabetic patients.

[0013] Native human amylin has the formula: H-KC()NTATC()ATQRLANFLVHSSNNFGAILSSTNVGSNTY-NH2 (SEQ ID NO: 3) is a peptide consisting of 37 amino acids having the formula In the formula, H- at the N-terminus indicates a hydrogen atom corresponding to the presence of a free amino group at the N-terminal amino acid residue (i.e., the lysine (K) residue at sequence position number 1 in the sequence shown above), -NH2 at the C-terminus indicates that the C-terminal carboxyl group is in amide form, and the parentheses "()" associated with the two cysteine ​​(C, Cys) residues at positions 2 and 7 of the sequence indicate the presence of an intramolecular disulfide bridge between the two Cys residues.

[0014] Amylin may be beneficial in the treatment of metabolic disorders such as diabetes and / or obesity. Amylin is thought to regulate gastric emptying and also suppress glucagon secretion and food intake, regulating the rate of glucose release into the circulation. Amylin is thought to complement the action of insulin. Compared to healthy adults, type 1 diabetic patients have no circulating amylin, and type 2 diabetic patients show low postprandial amylin concentrations. WO 93 / 10146 describes an amylin analog known as pramlintide, which has the sequence: Lys-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Asn-Phe-Leu-Val-His-Ser-Ser-Asn-Asn-Phe-Gly-Pro-Ile-Leu-Pro-Pro-Thr-Asn-Val-Gly-Ser-Asn-Thr-Tyr (SEQ ID NO: 4) It has.

[0015] Pramlintide also has a disulfide bridge between cysteine ​​residues at positions 2 and 7 and has been shown in human studies to reduce body weight or reduce weight gain.

[0016] Another amylin analogue, called IAPP-GI, which incorporates N-methylated residues and reduces its tendency to form fibrils, has been described by Yan et al. (PNAS, 103(7), 2046-2051, 2006; Angew. Chem. Int. Ed. 2013, 52, 10378-10383; WO 2006 / 042745). However, IAPP-GI appears to be less active than native amylin.

[0017] WO 2018 / 046719 describes amylin analogs that, among other things, have a lactam bridge instead of a disulfide bridge, an N-methylated residue, and deletions corresponding to residues Asn21 and Asn22 of native human amylin. Such analogs have a much lower tendency to fibrillate than native amylin, while also having higher potency than the analogs described by Yan et al. (supra). [Prior art documents] [Patent documents]

[0018] [Patent Document 1] International Publication No. 2013 / 164484 Brochure [Patent Document 2] International Publication No. 2018 / 104561 Brochure [Patent Document 3] International Application No. PCT / EP2022 / 074420 [Patent Document 4] International Publication No. 93 / 10146 Brochure [Patent Document 5] International Publication No. 2006 / 042745 Pamphlet [Patent Document 6] International Publication No. 2018 / 046719 Brochure [Non-patent literature]

[0019] [Non-Patent Document 1] Madsbad, S., 2014, Diabetes Obes Metab, 16: 9-21 [Non-patent document 2] Tolessa et al., 1998, Dig. Dis. Sci. 43(10): 2284-90 [Non-patent document 3] Filippatos et al, 2014 / 15, Rev Diabet Stud., 11(3): 202-230 [Non-patent document 4] Wilding et al, 2021, N Engl J Med; 384:989-1002 [Non-patent document 5] PNAS, 103(7), 2046-2051, 2006 [Non-patent document 6] Angew. Chem. Int. Ed. 2013, 52, 10378-10383 Summary of the Invention [Means for solving the problem]

[0020] Broadly, the present invention relates to therapies and methods for the prevention or treatment of diseases and disorders such as obesity and obesity-related conditions. More specifically, the present invention is based on the surprising discovery that the combined use of a GLP-1 (glucagon-like peptide 1) and GLP-2 (glucagon-like peptide 2) receptor dual agonist with a long-acting amylin analog results in superior therapeutic efficacy compared to treatment with either agent alone.

[0021] In one aspect, there is provided a GLP-1 / GLP-2 dual agonist and an amylin analogue for use in a method for preventing or treating a disease or disorder, wherein the GLP-1 / GLP-2 dual agonist and an amylin analogue are administered to a subject.

[0022] In one embodiment, the subject is a human subject.

[0023] In one embodiment, the subject is a human subject suffering from a disease or disorder.

[0024] In one embodiment, the subject is a human subject at risk of developing a disease or disorder.

[0025] In one embodiment, the dose of the GLP-1 / GLP-2 dual agonist and the dose of the amylin analog are independently selected.

[0026] In one embodiment, the GLP-1 / GLP-2 dual agonist may be administered at a dose of 100 nmol / kg and the amylin analog may be administered at a dose of 10 nmol / kg.

[0027] In one embodiment, the disease or disorder is obesity, morbid obesity, obesity-linked gallbladder disease, obesity-induced sleep apnea, inadequate glucose control, glucose tolerance, dyslipidemia, diabetes, pre-diabetes, metabolic syndrome, or hypertension.

[0028] The present invention further relates to the administration of a GLP-1 / GLP-2 dual agonist and an amylin analog for use in methods that are not themselves for the treatment of a disease or disorder. Such uses and methods can be considered for the prevention of a disease or disorder, provided they are used prior to the onset of the disease (e.g., prior to diagnosis), for example, to ameliorate an undesirable physiological characteristic or alter a particular physiological parameter.

[0029] In some embodiments, the uses or methods of the present invention can be considered cosmetic.

[0030] In one embodiment, the subject to whom the GLP-1 / GLP-2 dual agonist and amylin analog is administered is free of a disease or disorder.

[0031] In one aspect, there is provided a combination of a GLP-1 / GLP-2 dual agonist and an amylin analogue for use in a method of reducing or inhibiting weight gain, reducing gastric emptying or intestinal transit, reducing food intake, reducing appetite, or promoting weight loss, wherein the GLP-1 / GLP-2 dual agonist and amylin analogue are administered.

[0032] The present invention further provides a combination of a GLP-1 / GLP-2 dual agonist and an amylin analogue for use in therapy. In yet another aspect, there is provided a combination of a GLP-1 / GLP-2 dual agonist and an amylin analogue for use as a pharmaceutical. There is also provided a combination of a GLP-1 / GLP-2 dual agonist and an amylin analogue for use in a method of medical treatment.

[0033] The present invention also provides a combination of a GLP-1 / GLP-2 dual agonist and an amylin analogue for use in a method of reducing or inhibiting weight gain, reducing gastric emptying or intestinal transit, reducing food intake, reducing appetite, or promoting weight loss.

[0034] The present invention also provides a combination of a GLP-1 / GLP-2 dual agonist and an amylin analogue for use in a method for preventing or treating obesity, morbid obesity, obesity-related gallbladder disease, obesity-induced sleep apnea, poor glucose control, glucose tolerance, dyslipidemia (e.g., high LDL levels or low HDL / LDL ratio), diabetes (e.g., type 2 diabetes, gestational diabetes), pre-diabetes, metabolic syndrome, or hypertension.

[0035] The present invention also provides methods for reducing or inhibiting weight gain, reducing gastric emptying or intestinal transit, reducing food intake, reducing appetite, or promoting weight loss in a subject in need thereof, comprising administering to the subject a GLP-1 / GLP-2 dual agonist and an amylin analog.

[0036] The present invention also provides a method for preventing or treating obesity, morbid obesity, obesity-related gallbladder disease, obesity-induced sleep apnea, poor glucose control, glucose tolerance, dyslipidemia (e.g., high LDL levels or low HDL / LDL ratio), diabetes (e.g., type 2 diabetes, gestational diabetes), pre-diabetes, metabolic syndrome, or hypertension in a subject in need thereof, comprising administering to the subject a GLP-1 / GLP-2 dual agonist and an amylin analog.

[0037] The present invention also provides a GLP-1 / GLP-2 dual agonist and an amylin analog for use in preparing one or more medicaments for administration of the GLP-1 / GLP-2 dual agonist and the amylin analog. In this regard, the GLP-1 / GLP-2 dual agonist and the amylin analog may be in the same or different medicaments. In a preferred embodiment, the GLP-1 / GLP-2 dual agonist and the amylin analog may be in separate medicaments. In a preferred embodiment, the GLP-1 / GLP-2 dual agonist is in one medicament (sometimes referred to as a "GLP-1 / GLP-2 dual agonist medicament" or "dual agonist medicament" or "GLP-1 / GLP-2 dual agonist medicinal product" or "dual agonist medicinal product"), which may include one or more carriers, diluents, and excipients, and the amylin analog is in another medicament (sometimes referred to as an "amylin analog medicament" or "amylin analog medicinal product"), which may include one or more carriers, diluents, and excipients. The GLP-1 / GLP-2 dual agonist medicament can be in any form suitable for administration to a subject. In a preferred embodiment, the GLP-1 / GLP-2 dual agonist medicament is in a form suitable for subcutaneous (s / c or sc) administration to a subject. The amylin analog medicament can be in any form suitable for administration to a subject. In a preferred embodiment, the amylin analogue drug is in a form suitable for subcutaneous (s / c or sc) administration to a subject. The GLP-1 / GLP-2 dual agonist drug and the amylin analogue drug can be delivered to a subject at the same time or at different times. In a preferred embodiment, the GLP-1 / GLP-2 dual agonist drug is delivered to a subject daily. In a preferred embodiment, the GLP-1 / GLP-2 dual agonist drug is delivered to a subject once daily. In a preferred embodiment, the amylin analogue drug is delivered to a subject every other day. In a preferred embodiment, the amylin analogue drug is delivered to a subject once every other day.

[0038] In some embodiments, preferably, the methods comprise administering to the subject a GLP-1 / GLP-2 dual agonist at a dose of about 0.5 mg to about 10.0 mg.

[0039] In some embodiments, preferably, the methods comprise administering to the subject a GLP-1 / GLP-2 dual agonist at a dose of about 0.5 mg to about 7.5 mg, preferably about 1.0 mg to about 7.5 mg, preferably about 1.0 to about 6.0 mg, preferably about 1.0 to about 4.0 mg, preferably about 1.0 to about 3.5 mg.

[0040] In some embodiments, preferably, the methods comprise administering to the subject a GLP-1 / GLP-2 dual agonist once daily at a dose of about 0.5 mg to about 10.0 mg per day.

[0041] In some embodiments, preferably, the methods comprise administering a GLP-1 / GLP-2 dual agonist to the subject once daily at a dose of about 0.5 mg to about 7.5 mg per day, preferably once daily at a dose of about 1.0 mg to about 7.5 mg per day, preferably once daily at a dose of about 1.0 to about 6.0 mg per day, preferably once daily at a dose of about 1.0 to about 4.0 mg per day, preferably once daily at a dose of about 1.0 to about 3.5 mg per day.

[0042] In some embodiments, preferably the GLP-1 / GLP-2 dual agonist is in a pharmaceutical composition, the pharmaceutical composition comprising: (i) at least about 1 mg / mL of one or more GLP-1 / GLP-2 dual agonists (ii) 5 mM to about 50 mM of phosphate buffer components (iii) about 190 mM to about 240 mM mannitol, and / or (iv) The pH is approximately 8.0. Includes one or more of the following.

[0043] In some embodiments, preferably, the method comprises administering to the subject an amylin analog at a dose of about 0.5 mg to about 10.0 mg, preferably about 0.6 mg to about 7.5 mg, preferably about 1.2 mg to about 7.5 mg, preferably about 1.2 mg to about 6.0 mg, preferably about 2.4 mg to about 6.0 mg, preferably about 2.4 mg to about 4.0 mg, preferably about 2.4 mg to about 3.5 mg.

[0044] In some embodiments, the method preferably comprises administering an amylin analog to the subject at a dose of about 0.5 mg to about 10.0 mg per day every other day, preferably at a dose of about 0.6 mg to about 7.5 mg per day every other day, preferably at a dose of about 1.2 mg to about 7.5 mg per day every other day, preferably at a dose of about 1.2 to about 6.0 mg per day every other day, preferably at a dose of about 2.4 to about 6.0 mg per day every other day, preferably at a dose of about 2.4 to about 4.0 mg per day every other day, preferably at a dose of about 2.4 to about 3.5 mg per day every other day.

[0045] In some embodiments, preferably the amylin analog is in a pharmaceutical composition, the pharmaceutical composition comprising: (i) the one or more amylin analogs are at a concentration of about 0.4 mg / ml to about 25 mg / ml; (ii) being present in a solution having a buffer concentration of about 0.5 mM to 25 mM; (iii) in a solution having a pH of about 5.8 to about 6.9; and / or (iv) Provided as a chloride salt. Includes one or more of the following.

[0046] In some embodiments, preferably, the method comprises administering a GLP-1 / GLP-2 dual agonist to the subject once daily at a dose of about 0.5 mg to about 7.5 mg per day, preferably once daily at a dose of about 1.0 mg to about 7.5 mg per day, preferably once daily at a dose of about 1.0 to about 6.0 mg per day, preferably once daily at a dose of about 1.0 to about 4.0 mg per day, preferably once daily at a dose of about 1.0 to about 3.5 mg per day; Here, the method includes administering an amylin analog to a subject at a dose of about 0.5 mg to about 10.0 mg per day every other day, preferably at a dose of about 0.6 mg to about 7.5 mg per day every other day, preferably at a dose of about 1.2 mg to about 7.5 mg per day every other day, preferably at a dose of about 1.2 to about 6.0 mg per day every other day, preferably at a dose of about 2.4 to about 6.0 mg per day every other day, preferably at a dose of about 2.4 to about 4.0 mg per day every other day, preferably at a dose of about 2.4 to about 3.5 mg per day every other day.

[0047] In some embodiments, preferably, the method comprises administering a GLP-1 / GLP-2 dual agonist to the subject once daily at a dose of about 0.5 mg to about 7.5 mg per day, preferably once daily at a dose of about 1.0 mg to about 7.5 mg per day, preferably once daily at a dose of about 1.0 to about 6.0 mg per day, preferably once daily at a dose of about 1.0 to about 4.0 mg per day, preferably once daily at a dose of about 1.0 to about 3.5 mg per day; the GLP-1 / GLP-2 dual agonist is in a first pharmaceutical composition; The first pharmaceutical composition comprises: (i) at least about 1 mg / mL of one or more GLP-1 / GLP-2 dual agonists (ii) 5 mM to about 50 mM of phosphate buffer components (iii) about 190 mM to about 240 mM mannitol, and / or (iv) The pH is approximately 8.0. wherein the method comprises administering to the subject an amylin analog at a dose of about 0.5 mg to about 10.0 mg per day every other day, preferably at a dose of about 0.6 mg to about 7.5 mg per day every other day, preferably at a dose of about 1.2 mg to about 7.5 mg per day every other day, preferably at a dose of about 1.2 to about 6.0 mg per day every other day, preferably at a dose of about 2.4 to about 6.0 mg per day every other day, preferably at a dose of about 2.4 to about 4.0 mg per day every other day, preferably at a dose of about 2.4 to about 3.5 mg per day every other day; and The amylin analog is in a second pharmaceutical composition, the second pharmaceutical composition comprising: (i) the one or more amylin analogs are at a concentration of about 0.4 mg / ml to about 25 mg / ml; (ii) being present in a solution having a buffer concentration of about 0.5 mM to 25 mM; (iii) in a solution having a pH of about 5.8 to about 6.9; and / or (iv) the amylin analog is provided as a chloride salt; Includes one or more of the following.

[0048] In some embodiments, preferably, the method comprises administering a GLP-1 / GLP-2 dual agonist to the subject once daily at a dose of about 0.5 mg to about 7.5 mg per day, preferably once daily at a dose of about 1.0 mg to about 7.5 mg per day, preferably once daily at a dose of about 1.0 to about 6.0 mg per day, preferably once daily at a dose of about 1.0 to about 4.0 mg per day, preferably once daily at a dose of about 1.0 to about 3.5 mg per day; the GLP-1 / GLP-2 dual agonist is in a first pharmaceutical composition; The first pharmaceutical composition comprises: (i) at least about 1 mg / mL of one or more GLP-1 / GLP-2 dual agonists (ii) 5 mM to about 50 mM of phosphate buffer components (iii) about 190 mM to about 240 mM mannitol, and / or (iv) The pH is approximately 8.0. wherein the method comprises administering to the subject an amylin analog at a dose of about 0.5 mg to about 10.0 mg per day every other day, preferably at a dose of about 0.6 mg to about 7.5 mg per day every other day, preferably at a dose of about 1.2 mg to about 7.5 mg per day every other day, preferably at a dose of about 1.2 to about 6.0 mg per day every other day, preferably at a dose of about 2.4 to about 6.0 mg per day every other day, preferably at a dose of about 2.4 to about 4.0 mg per day every other day, preferably at a dose of about 2.4 to about 3.5 mg per day every other day; and The amylin analog is in a second pharmaceutical composition, the second pharmaceutical composition comprising: (i) the one or more amylin analogs are at a concentration of about 0.4 mg / ml to about 25 mg / ml; (ii) being present in a solution having a buffer concentration of about 0.5 mM to 25 mM; (iii) in a solution having a pH of about 5.8 to about 6.9; and / or (iv) the amylin analog is provided as a chloride salt; Includes.

[0049] The present invention also provides the use of a GLP-1 / GLP-2 dual agonist and an amylin analogue in the preparation of a medicament for reducing or inhibiting weight gain, reducing gastric emptying or intestinal transit, reducing food intake, reducing appetite, or promoting weight loss.

[0050] The present invention also provides use of a GLP-1 / GLP-2 dual agonist and an amylin analog in the preparation of a medicament for preventing or treating obesity, morbid obesity, obesity-related gallbladder disease, obesity-induced sleep apnea, poor glucose control, glucose tolerance, dyslipidemia (e.g., high LDL levels or low HDL / LDL ratio), diabetes (e.g., type 2 diabetes, gestational diabetes), pre-diabetes, metabolic syndrome, or hypertension.

[0051] A further aspect provides a therapeutic kit comprising a dual agonist or a pharmaceutically acceptable salt or solvate thereof according to the invention.

[0052] The methods and uses of the present invention may be carried out in vitro, in vivo or ex vivo.

[0053] Compounds having agonist activity at the GLP-1 (glucagon-like peptide 1) receptor and the GLP-2 (glucagon-like peptide 2) receptor (e.g., as assessed in an in vitro potency assay). Such compounds are referred to herein as "GLP-1 / GLP-2 dual agonists," or simply "dual agonists." Thus, the compounds of the present invention have both GLP-1 and GLP-2 activity.

[0054] In one embodiment, the GLP-1 / GLP-2 dual agonist is Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecanoyl]-isoGlu)]QAARDFIAWLIQHKITD-OH (SEQ ID NO: 5) is.

[0055] The dual agonist may be in the form of a pharmaceutically acceptable salt or solvate, such as a pharmaceutically acceptable acid addition salt.

[0056] The present invention also provides a composition comprising the dual agonist of the present invention or a pharmaceutically acceptable salt or solvate thereof together with a carrier, excipient, or vehicle. The carrier may be a pharmaceutically acceptable carrier. The composition may be a pharmaceutical composition. The pharmaceutical composition may be formulated as a liquid suitable for administration by injection or infusion.

[0057] In one embodiment, the amylin analog is [19CD]-isoGlu-RD()GTATK()ATERLA-Aad-FLQRSSF-Gly(Me)-A-Ile(Me)-LSSTEVGSNT-Hyp-NH2 (SEQ ID NO: 6) is.

[0058] The amylin analogue may be in the form of a pharmaceutically acceptable salt or solvate, for example a pharmaceutically acceptable acid addition salt.

[0059] The present invention also provides a composition comprising the amylin analog of the present invention or a pharmaceutically acceptable salt or solvate thereof together with a carrier, excipient, or vehicle. The carrier can be a pharmaceutically acceptable carrier. The composition can be a pharmaceutical composition. The pharmaceutical composition can be formulated as a liquid suitable for administration by injection or infusion. It can be formulated to achieve delayed release of the dual agonist. [Brief explanation of the drawings]

[0060] [Figure 1] Figure 1 shows the effect of sc treatment with vehicle, GLP-1 / GLP-2 dual agonist, amylin analog, and combination therapy on cumulative food intake in diet-induced obese (DIO) Sprague-Dawley rats. Data are shown as mean values ​​(n=9 / group). Statistics: Cumulative food intake on day 28 (days 0-28) was compared by one-way ANOVA followed by Bonferroni's multiple comparison test with 95% confidence intervals. [Figure 2]Figure 1 shows the effect of sc treatment with vehicle, GLP-1 / GLP-2 dual agonist or amylin analog monotherapy, and GLP-1 / GLP-2 dual agonist and amylin analog combination therapy on % body weight (BW) change from baseline, day 0, in diet-induced obese (DIO) Sprague-Dawley rats. Data are shown as mean values ​​(n=9 / group). A) Daily change in BW %. B) Change in BW at day 28. Statistics: Data in B) were compared by one-way ANOVA followed by Bonferroni's multiple comparison test with 95% confidence intervals. [Figure 3]

[0033] Figure 1 shows the effect of subcutaneous (sc) treatment with vehicle, GLP-1 / GLP-2 dual agonist or amylin analog monotherapy, and GLP-1 / GLP-2 dual agonist and amylin analog combination therapy on overnight fasted blood glucose levels in diet-induced obese (DIO) Sprague-Dawley rats on day 29. Data are presented as mean values ​​(n=9 / group). The dotted line indicates the mean baseline fasted blood glucose levels obtained from all animals in the study. Statistics: Data were compared by one-way ANOVA followed by Bonferroni's multiple comparison test with 95% confidence intervals. [Figure 4] Figure 1 shows plasma exposure in nmol / L measured using LC-MS / MS for: A. GLP-1 / GLP-2 dual agonist at a dose of 100 nmol / kg, either individually or in combination with 10 nmol / kg amylin analogue. B. Amylin analogue at a dose of 10 nmol / kg, either individually or in combination with 100 nmol / kg GLP-1 / GLP-2 dual agonist. Data are shown as mean values ​​(n=9 / group) and error bars represent 1 standard deviation. DETAILED DESCRIPTION OF THE INVENTION

[0061] Unless otherwise defined herein, scientific and technical terms used in this application have the meanings commonly understood by those of ordinary skill in the art. Overall, the nomenclature used in connection with, and techniques of, chemistry, molecular biology, cell and cancer biology, immunology, microbiology, pharmacology, and protein and nucleic acid chemistry described herein are those well known and commonly used in the art.

[0062] All patents, published patent applications, and non-patent publications referenced in this application are specifically incorporated herein by reference. In case of conflict, the present specification, including its specific definitions, will control.

[0063] Each embodiment of the invention described herein may be practiced alone or in combination with one or more other embodiments of the invention.

[0064] definition Unless otherwise specified, the following definitions are given to specific terms used herein.

[0065] Throughout this specification, the word "comprise" and grammatical variations thereof, such as "comprises" or "comprising," will be understood to mean the inclusion of a stated integer or component or group of integers or components, but not the exclusion of any other integer or component or group of integers or components.

[0066] The singular forms "a," "an," and "the" include the plural forms unless the context clearly indicates otherwise.

[0067] The term "including" is used to mean "including but not limited to." "Including" and "including but not limited to" can be used interchangeably.

[0068] The terms "patient," "subject," and "individual" can be used interchangeably and refer to either a human or a non-human animal. These terms include mammals, such as humans, primates, livestock animals (e.g., cows and pigs), companion animals (e.g., dogs and cats), and rodents (e.g., mice and rats).

[0069] The term "solvate" in the context of the present invention refers to a complex with a defined stoichiometry formed between a solute (in this case, a peptide according to the invention or a pharmaceutically acceptable salt thereof) and a solvent. The solvent in this context can be, for example, water, ethanol, or another pharmaceutically acceptable, typically low-molecular-weight organic species, such as, but not limited to, acetic acid or lactic acid. When the solvent in question is water, such a solvate is usually called a hydrate.

[0070] The term "agonist" as used in the context of the present invention refers to a substance (ligand) that activates the receptor type in question.

[0071] Throughout this specification and claims, the conventional three-letter and one-letter codes for natural amino acids are used, i.e., A(Ala), G(Gly), L(Leu), I(Ile), V(Val), F(Phe), W(Trp), S(Ser), T(Thr), Y(Tyr), N (Asn), Q(Gln), D(Asp), E(Glu), K(Lys), R(Arg), H(His), M(Met), C(Cys), and P(Pro), and commonly accepted three-letter codes for other alpha amino acids, such as sarcosine (Sar), norleucine (Nle), alpha-aminoisobutyric acid (Aib), 2,3-diaminopropanoic acid (Dap), 2,4-diaminobutanoic acid (Dab), and 2,5-diaminopentanoic acid (ornithine, Orn). Such other alpha amino acids, when used in general formulas or sequences herein, may be indicated in square brackets "[]" (e.g., "[Aib]"), particularly when the remainder of the formula or sequence is shown using single-letter codes. Unless otherwise specified, amino acid residues in the peptides of the invention are in the L-configuration. However, D-configuration amino acids may also be incorporated. In this context, amino acid codes written in lowercase represent the D-configuration of the amino acid, e.g., "k" represents the D-configuration of lysine (K).

[0072] Some sequences disclosed herein incorporate a "Hy-" moiety at the amino-terminus (N-terminus) of the sequence and either an "-OH" or an "-NH2" moiety at the carboxy-terminus (C-terminus) of the sequence. In such cases, and unless otherwise indicated, the "Hy-" moiety at the N-terminus of the sequence in question is a hydrogen atom [i.e., in the general formula R 1 = hydrogen = Hy; corresponds to the presence of a free primary or secondary amino group at the N-terminus], while an "-OH" or "-NH" moiety at the C-terminus of a sequence indicates a hydroxy group [e.g., R 2 =OH; corresponding to the presence of a carboxy (COOH) group at the C-terminus] or an amino group [e.g., in the general formula R 2 = [NH2]; corresponding to the presence of an amide (CONH2) group at the C-terminus]. In each sequence of the present invention, a C-terminal "-OH" moiety can be substituted for a C-terminal "-NH2" moiety, and vice versa.

[0073] "Percent (%) amino acid sequence identity" with respect to a GLP-2 polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to those in the wild-type (human) GLP-2 sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity; any conservative substitutions are not considered part of the sequence identity. Sequence alignment can be performed by those skilled in the art using techniques well known in the art, for example, using publicly available software such as BLAST, BLAST2, or Align software. See, for example, Altschul et al., Methods in Enzymology 266: 460-480 (1996), or Pearson et al., Genomics 46: 24-36, 1997.

[0074] As used herein in the context of the present invention, the percentage of sequence identity can be determined using these programs at their default settings. More generally, those skilled in the art can readily determine appropriate parameters for determining alignment, including any algorithms needed to obtain maximal alignment over the full length of the sequences being compared.

[0075] Dual agonist compounds Compounds having agonist activity at the GLP-1 (glucagon-like peptide 1) receptor and the GLP-2 (glucagon-like peptide 2) receptor (e.g., as assessed in an in vitro potency assay). Such compounds are referred to herein as "GLP-1 / GLP-2 dual agonists" or "dual agonists." Thus, the compounds of the present invention have both GLP-1 and GLP-2 activity.

[0076] The dual agonist compounds according to the present invention have both GLP-1(7-36) and GLP-2(1-33) activity.

[0077] The GLP-1 / GLP-2 dual agonists of the present invention have the formula: R1 -X * -UR 2 or a pharmaceutically acceptable salt or solvate thereof, During the ceremony: -R 1 is hydrogen (Hy), C 1-4 alkyl (e.g., methyl), acetyl, formyl, benzoyl, or trifluoroacetyl; -R 2 is NH or OH, - X * is represented by Formula I: H-X2-EG-X5-F-X7-X8-E-X10-X11-TIL-X15-X16-X17-A-X19-X20-X21-FI-X24-WL-X27-X28-X29-KIT-X33(I) (Sequence number 7) is a peptide of During the ceremony, - X2 is Aib or G, - X5 is T or S; - X7 is T or S; - X8 is S, E, or D; X10 is L, M, V or Ψ; X11 is A, N or S; - X15 is D or E, X16 is G, E, A or Ψ, X17 is Q, E, K, L or Ψ, - X19 is A, V or S; X20 is R, K or Ψ, X21 is D, L or E; X24 is A, N, or S; X27 is I, Q, K, H or Y; X28 is Q, E, A, H, Y, L, K, R, or S; - X29 is H, Y, K or Q; - X33 is D or E, - U is absent or a sequence consisting of 1 to 15 residues independently selected from K, k, E, A, T, I, L, and Ψ; - the molecule has one or only one Ψ, which is a group of formula Z 1 - or Z 1 -Z 2 a residue K, k, R, Orn, Dap, or Dab, the side chain of which is conjugated to a substituent having the formula -Z 1 - is CH3-(CH2) 10-22 -(CO)- or HOOC-(CH2) 10-22 -(CO)-, and - -Z 2 -Z S1 -, -Z S1 -Z S2 -, -Z S2 -Z S1 , -Z S2 -, -Z S3 -, -Z S1 Z S3 -, -Z S2 Z S3 -, -Z S3 Z S1 -, -Z S3 Z S2 -, -Z S1 Z S2 Z S3 -, -Z S1 Z S3 Z S2 -, -Z S2 Z S1 Z S3 -, -Z S2 Z S3 Z S1 -, -Z S3 Z S1 Z S2 -, -Z S3 Z S2 Z S1 -, -Z S2 Z S3 Z S2 - selected from, where: -Z S1 is isoGlu, β-Ala, isoLys, or 4-aminobutanoyl; -Z S2 Ha-(Peg3)m where m is 1, 2, or 3, and - -Z S3 - is a peptide sequence consisting of 1 to 6 amino acid units independently selected from the group consisting of A, L, S, T, Y, Q, D, E, K, k, R, H, F, and G; and at least one of X5 and X7 is T.

[0078] In one embodiment, the GLP-1 / GLP-2 dual agonist is Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecanoyl]-isoGlu)]QAARDFIAWLIQHKITD-OH (SEQ ID NO: 5) is.

[0079] The dual agonist may be in the form of a pharmaceutically acceptable salt or solvate, such as a pharmaceutically acceptable acid addition salt.

[0080] The present invention also provides a composition comprising the dual agonist of the present invention or a pharmaceutically acceptable salt or solvate thereof together with a carrier, excipient, or vehicle. The carrier may be a pharmaceutically acceptable carrier. The composition may be a pharmaceutical composition. The pharmaceutical composition may be formulated as a liquid suitable for administration by injection or infusion.

[0081] Dual agonist activity According to the present invention, a dual agonist has at least one GLP-1 biological activity and at least one GLP-2 biological activity. Exemplary GLP-1 physiological activities include reducing intestinal transit rate, reducing gastric emptying rate, reducing appetite, food intake, or body weight, and improving glucose control and glucose tolerance. Exemplary GLP-2 physiological activities include increasing intestinal bulk (e.g., in the small intestine or colon), repairing the intestinal tract, and improving intestinal barrier function (i.e., reducing intestinal permeability). These parameters can be assessed in in vivo assays that determine the bulk and permeability of the intestinal tract or a portion thereof after treating a test animal with a dual agonist.

[0082] Dual agonists have agonist activity at the GLP-1 receptor and the GLP-2 receptor, e.g., the human GLP-1 receptor and the GLP-2 receptor. EC 50 The EC value can be used as a numerical measure of agonist potency at a given receptor. 50 The EC value is a measure of the compound concentration (e.g., mol / L) required to achieve half of the maximal activity of that compound in a particular assay. The numerical EC value at a particular receptor in the same assay 50 is the EC of the reference compound 50 A compound lower than the reference compound can be considered to be more potent at that receptor than the reference compound.

[0083] GLP-1 activity In some embodiments, the dual agonist has an EC50 at a GLP-1 receptor (e.g., a human GLP-1 receptor) of less than 2.0 nM, less than 1.5 nM, less than 1.0 nM, less than 0.9 nM, less than 0.8 nM, less than 0.7 nM, less than 0.6 nM, less than 0.5 nM, less than 0.4 nM, less than 0.3 nM, less than 0.2 nM, less than 0.1 nM, less than 0.09 nM, less than 0.08 nM, less than 0.07 nM, less than 0.06 nM, less than 0.05 nM, or less than 0.04 nM, as assessed using, for example, the GLP-1 receptor potency assay described in International Application No. PCT / EP2022 / 074420. 50It has.

[0084] In some embodiments, the dual agonist has a potency of 0.005-2.5 nM, 0.01 nM-2.5 nM, 0.025-2.5 nM, 0.005-2.0 nM, 0.01 nM-2.0 nM, 0.025-2.0 nM, 0.005-2.0 nM, 0.01 nM-2.0 nM, 0.025-2.0 nM, 0.00 EC values ​​at the GLP-1 receptor of 5-1.5nM, 0.01nM-1.5nM, 0.025-1.5nM, 0.005-1.0nM, 0.01nM-1.0nM, 0.025-1.0nM, 0.005-0.5nM, 0.01nM-0.5nM, 0.025-0.5nM, 0.005-0.25nM, 0.01nM-0.25nM, and 0.025-0.25nM 50 It has.

[0085] Another measure of GLP-1 agonist activity can be derived by comparing the potency of the dual agonist with that of a known (or reference) GLP-1 agonist when both are measured in the same assay. Thus, relative potency at the GLP-1 receptor can be defined as: [EC 50 (reference agonist)] / [EC 50 (dual agonist)].

[0086] Thus, a value of 1 indicates that the dual agonist and reference agonist have equal potency, and a value greater than 1 indicates that the dual agonist has greater potency (i.e., a lower EC 50 ), and a value less than 1 indicates that the dual agonist has lower potency (i.e., a higher EC 50 ) is indicated.

[0087] The reference GLP-1 agonist can be, for example, human GLP-1(7-37), liraglutide (NN2211, Victoza), or exendin-4, but is preferably liraglutide.

[0088] Typically, the relative potency will be between 0.001 and 100, for example, between 0.001 and 10, between 0.001 and 5, between 0.001 and 1, between 0.001 and 0.5, between 0.001 and 0.1, between 0.001 and 0.05, or between 0.001 and 0.01; between 0.01 and 10, between 0.01 and 5, between 0.01 and 1, between 0.01 and 0.5, between 0.01 and 0.1, or between 0.01 and 0.05; between 0.05 and 10, between 0.05 and 5, between 0.05 and 1, between 0.05 and 0.5, or between 0.05 and 0.1; between 0.1 and 10, between 0.1 and 5, between 0.1 and 1, or between 0.1 and 0.5; between 0.5 and 10, between 0.5 and 5, or between 0.5 and 1; between 1 and 10 or between 1 and 5; or between 5 and 10.

[0089] The dual agonists described in the Examples below have slightly lower GLP-1 potencies than liraglutide and so may have relative potencies of, for example, 0.01-1, 0.01-0.5, or 0.01-0.1.

[0090] Conversely, the dual agonists of the invention have greater potency at the GLP-1 receptor (e.g., the human GLP-1 receptor) than wild-type human GLP-2 (hGLP-2(1-33)) or [Gly2]-hGLP-2(1-33) (i.e., human GLP-2 with glycine at position 2, also known as teduglutide). Thus, the relative potency of the dual agonist at the GLP-1 receptor compared to hGLP-2(1-33) or teduglutide is greater than 1, typically greater than 5 or greater than 10, and can be up to 100, up to 500, or even higher.

[0091] GLP-2 activity In some embodiments, the dual agonist has an EC50 at a GLP-2 receptor (e.g., a human GLP-2 receptor) of less than 2.0 nM, less than 1.5 nM, less than 1.0 nM, less than 0.9 nM, less than 0.8 nM, less than 0.7 nM, less than 0.6 nM, less than 0.5 nM, less than 0.4 nM, less than 0.3 nM, less than 0.2 nM, less than 0.1 nM, less than 0.09 nM, less than 0.08 nM, less than 0.07 nM, less than 0.06 nM, less than 0.05 nM, less than 0.04 nM, less than 0.03 nM, less than 0.02 nM, or less than 0.01 nM, as assessed using, for example, the GLP-2 receptor potency assay described in International Application No. PCT / EP2022 / 074420. 50 It has.

[0092] In some embodiments, the dual agonist has an EC50 at the GLP-2 receptor of 0.005 to 2.0 nM, 0.01 nM to 2.0 nM, 0.025 to 2.0 nM, 0.005 to 1.5 nM, 0.01 nM to 1.5 nM, 0.025 to 1.5 nM, 0.005 to 1.0 nM, 0.01 nM to 1.0 nM, 0.025 to 1.0 nM, 0.005 to 0.5 nM, 0.01 nM to 0.5 nM, 0.025 to 0.5 nM, 0.005 to 0.25 nM, 0.01 nM to 0.25 nM, or 0.025 to 0.25 nM, as assessed using, for example, the GLP-2 receptor potency assay described in International Application No. PCT / EP2022 / 074420. 50 It has.

[0093] Another measure of GLP-2 agonist activity can be derived by comparing the potency of the dual agonist with that of a known (or reference) GLP-2 agonist when both are measured in the same assay. Thus, relative potency at the GLP-2 receptor can be defined as: [EC 50 (reference agonist)] / [EC 50 (dual agonist)].

[0094] Thus, a value of 1 indicates that the dual agonist and reference agonist have equal potency, and a value greater than 1 indicates that the dual agonist has greater potency (i.e., a lower EC 50 ), and a value less than 1 indicates that the dual agonist has lower potency (i.e., a higher EC 50 ) is indicated.

[0095] The reference GLP-2 agonist can be, for example, human GLP-2(1-33) or teduglutide ([Gly2]-hGLP-2(1-33)), but is preferably teduglutide. Typically, the relative potency will be between 0.001 and 100, for example, between 0.001 and 10, between 0.001 and 5, between 0.001 and 1, between 0.001 and 0.5, between 0.001 and 0.1, between 0.001 and 0.05, or between 0.001 and 0.01; between 0.01 and 10, between 0.01 and 5, between 0.01 and 1, between 0.01 and 0.5, between 0.01 and 0.1, or between 0.01 and 0.05; between 0.05 and 10, between 0.05 and 5, between 0.05 and 1, between 0.05 and 0.5, or between 0.05 and 0.1; between 0.1 and 10, between 0.1 and 5, between 0.1 and 1, or between 0.1 and 0.5; between 0.5 and 10, between 0.5 and 5, or between 0.5 and 1; between 1 and 10 or between 1 and 5; or between 5 and 10. The dual agonists described in the Examples below have slightly lower GLP-2 potencies than teduglutide, and so may have relative potencies of, for example, 0.01-1, 0.01-0.5, or 0.01-0.1.

[0096] Conversely, the dual agonists of the present invention have greater potency at the GLP-2 receptor (e.g., the human GLP-2 receptor) than human GLP-1(7-37), liraglutide (NN2211, Victoza), or exendin-4. Thus, the relative potency of the dual agonist at the GLP-2 receptor compared to human GLP-1(7-37), liraglutide (NN2211, Victoza), or exendin-4 is greater than 1, typically greater than 5 or greater than 10, and may be up to 100, up to 500, or even higher (if the reference GLP-1 agonist also exhibits detectable activity at the GLP-2 receptor).

[0097] It will be understood that the absolute potency of a dual agonist at each receptor is not as important as the balance between GLP-1 and GLP-2 agonist activity. Thus, as long as a dual agonist compound exhibits acceptable relative potency levels at both receptors, it is not a problem if the absolute potency of GLP-1 or GLP-2 at these receptors is lower than that of known agonists. Any apparent deficiency in absolute potency can be compensated for by increasing the dose, if necessary.

[0098] Pharmaceutical Composition One aspect of the present invention relates to a composition comprising a dual agonist according to the present invention or a pharmaceutically acceptable salt or solvate thereof together with a carrier. In one embodiment of the present invention, the composition is a pharmaceutical composition, and the carrier is a pharmaceutically acceptable carrier. The present invention also relates to a pharmaceutical composition comprising a dual agonist according to the present invention or a salt or solvate thereof together with a carrier, excipient, or vehicle. Thus, the dual agonist of the present invention or a salt or solvate thereof, particularly a pharmaceutically acceptable salt or solvate thereof, can be formulated as a composition or pharmaceutical composition prepared for storage or administration, which contains a therapeutically effective amount of the dual agonist of the present invention or a salt or solvate thereof.

[0099] Suitable salts formed with bases include metal salts, such as alkali metal salts or alkaline earth metal salts, for example, sodium salts, potassium salts, or magnesium salts; ammonia salts, and organic amine salts, such as those formed with morpholine, thiomorpholine, piperidine, pyrrolidine, lower mono-, di-, or tri-alkylamines (e.g., ethyl, tert-butyl, diethyl, diisopropyl, triethyl, tributyl, or dimethylpropylamine), or lower mono-, di-, or tri-(hydroxyalkyl)amines (e.g., mono-, di-, or tri-ethanolamine). Internal salts can also be formed. Similarly, when the compound of the present invention contains a basic moiety, salts can be formed using organic or inorganic acids. For example, salts can be formed with the following acids: formic acid, acetic acid, propionic acid, butyric acid, valeric acid, caproic acid, oxalic acid, lactic acid, citric acid, tartaric acid, succinic acid, fumaric acid, maleic acid, malonic acid, mandelic acid, malic acid, phthalic acid, hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, sulfuric acid, benzoic acid, carbonic acid, uric acid, methanesulfonic acid, naphthalenesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, p-toluenesulfonic acid (i.e., 4-methylbenzenesulfonic acid), camphorsulfonic acid, 2-aminoethanesulfonic acid, aminomethylsulfonic acid, and trifluoromethanesulfonic acid (the latter also referred to as triflic acid), as well as other known pharmaceutically acceptable acids. Amino acid addition salts can also be formed with amino acids such as lysine, glycine, or phenylalanine.

[0100] In one embodiment, the pharmaceutical composition of the present invention is one in which the dual agonist is in the form of a pharmaceutically acceptable acid addition salt.

[0101] In some embodiments, the pharmaceutical compositions of the invention are formulated as a 1 mL injectable solution.

[0102] In some embodiments, the pharmaceutical composition of the present invention comprises: (i) at least about 1 mg / mL of one or more GLP-1 / GLP-2 dual agonists (ii) 5 mM to about 50 mM of phosphate buffer components (iii) about 190 mM to about 240 mM mannitol and has a pH of approximately pH 8.0.

[0103] In some aspects, the compositions of the present invention comprise the following listed components: [Table 1] wherein the GLP-1 / GLP-2 dual agonist is a pharmaceutically acceptable salt of Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecanoyl]-isoGlu)]QAARDFIAWLIQHKITD-OH (SEQ ID NO: 5).

[0104] In some aspects, the compositions of the present invention comprise the following listed components: [Table 2] wherein the GLP-1 / GLP-2 dual agonist is a pharmaceutically acceptable salt of Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecanoyl]-isoGlu)]QAARDFIAWLIQHKITD-OH (SEQ ID NO: 5).

[0105] Dosage regime It will be understood that the dose of the GLP-1 / GLP-2 dual agonist and the dose of the amylin analogue may be selected independently. The dose of the GLP-1 / GLP-2 dual agonist may be selected as described herein.

[0106] In accordance with the present invention, a dual GLP-1 / GLP-2 agonist is for use in a method for reducing or inhibiting weight gain, reducing food intake, reducing appetite, promoting weight loss, or treating obesity, morbid obesity, obesity-related gallbladder disease, or obesity-induced sleep apnea. The invention includes a method of administering the dual agonist to a patient at a dose of about 0.1 mg to 10.0 mg.

[0107] A dose of about 0.1 mg to 10.0 mg of the dual agonist is administered to the patient in a single administration (i.e., a single administration event). In other words, the dual agonist is administered to the patient in a single dosage formulation of about 0.1 mg to about 10.0 mg. This single dosage formulation can be administered to the patient one or more times, in which case each of the multiple dosage formulations administered to the patient need not contain the same amount of the dual agonist. In other words, the dual agonist can be administered to the patient in a series of single dosages, in which each single dosage may not contain the same amount of the dual agonist. Each administration of the dual agonist to the patient can be independently selected to result in a dose of about 0.1 mg to about 10.0 mg.

[0108] Accordingly, the present invention provides a GLP-1 / GLP-2 dual agonist, or a pharmaceutically acceptable salt or solvate thereof, as described herein, for use in a method of reducing or inhibiting weight gain, reducing food intake, reducing appetite, promoting weight loss, or treating obesity, morbid obesity, obesity-related gallbladder disease, or obesity-induced sleep apnea, comprising at least one administration of the dual agonist to a patient at a dose of about 0.1 mg to 10.0 mg.

[0109] dose In one embodiment, the dual agonist is administered to a patient at a dose of about 0.1 mg to about 10.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 1.0 mg to about 10.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 1.1 mg to about 10.0 mg, about 1.2 mg to about 10.0 mg, about 1.3 mg to about 10.0 mg, or about 1.4 mg to about 10.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 1.5 mg to about 10.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 1.6 mg to about 10.0 mg, about 1.7 mg to about 10.0 mg, about 1.8 mg to about 10.0 mg, or about 1.9 mg to about 10.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 2.0 mg to about 10.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 2.1 mg to about 10.0 mg or about 2.2 mg to about 10.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 2.25 mg to about 10.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 3.0 mg to about 10.0 mg, about 4.0 mg to about 10.0 mg, about 5.0 mg to about 10.0 mg, about 6.0 mg to about 10.0 mg, about 7.0 mg to about 10.0 mg, about 8.0 mg to about 10.0 mg, or about 9.0 mg to about 10.0 mg.

[0110] In one embodiment, the dual agonist is administered to a patient at a dose of about 0.1 mg to about 9.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 1.0 mg to about 9.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 1.1 mg to about 9.0 mg, about 1.2 mg to about 9.0 mg, about 1.3 mg to about 9.0 mg, or about 1.4 mg to about 9.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 1.5 mg to about 9.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 1.6 mg to about 9.0 mg, about 1.7 mg to about 9.0 mg, about 1.8 mg to about 9.0 mg, or about 1.9 mg to about 9.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 2.0 mg to about 9.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 2.1 mg to about 9.0 mg or about 2.2 mg to about 9.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 2.25 mg to about 9.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 3.0 mg to about 9.0 mg, about 4.0 mg to about 9.0 mg, about 5.0 mg to about 9.0 mg, about 6.0 mg to about 9.0 mg, about 7.0 mg to about 9.0 mg, or about 8.0 mg to about 9.0 mg.

[0111] In one embodiment, the dual agonist is administered to a patient at a dose of about 0.1 mg to about 8.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 1.0 mg to about 8.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 1.1 mg to about 8.0 mg, about 1.2 mg to about 8.0 mg, about 1.3 mg to about 8.0 mg, or about 1.4 mg to about 8.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 1.5 mg to about 8.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 1.6 mg to about 8.0 mg, about 1.7 mg to about 8.0 mg, about 1.8 mg to about 8.0 mg, or about 1.9 mg to about 8.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 2.0 mg to about 8.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 2.1 mg to about 8.0 mg or about 2.2 mg to about 8.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 2.25 mg to about 8.0 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 3.0 mg to about 8.0 mg, about 4.0 mg to about 8.0 mg, about 5.0 mg to about 8.0 mg, about 6.0 mg to about 8.0 mg, or about 7.0 mg to about 8.0 mg.

[0112] In one embodiment, the dual agonist is administered to a patient at a dose of about 1.0 mg to about 7.5 mg, about 1.0 mg to about 7.0 mg, about 1.0 mg to about 6.0 mg, about 1.0 mg to about 5.0 mg, about 1.0 mg to about 4.0 mg, or about 1.0 mg to about 3.5 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 1.5 mg to about 7.5 mg. In another embodiment, the dual agonist is administered to a patient at a dose of about 1.5 mg to about 7.0 mg, about 1.5 mg to about 6.0 mg, about 1.5 mg to about 5.0 mg, about 1.5 mg to about 4.0 mg, or about 1.5 mg to about 3.5 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 2.0 mg to about 7.5 mg, about 2.0 mg to about 7.0 mg, about 2.0 mg to about 6.0 mg, about 2.0 mg to about 5.0 mg, about 2.0 mg to about 4.0 mg, or about 2.0 mg to about 3.5 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 2.25 mg to about 7.5 mg, about 2.25 mg to about 7.0 mg, about 2.25 mg to about 6.0 mg, about 2.25 mg to about 5.0 mg, about 2.25 mg to about 4.0 mg, or about 2.25 mg to about 3.5 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 4.0 mg to about 7.5 mg. In one embodiment, the dual agonist is administered to a patient at a dose of about 4.0 mg to about 6.0 mg.

[0113] In one embodiment, the dual agonist is administered to a patient at a dose of 1.0 mg to 7.5 mg, 1.0 mg to 7.0 mg, 1.0 mg to 6.0 mg, 1.0 mg to 5.0 mg, 1.0 mg to 4.0 mg, or 1.0 mg to 3.5 mg. In one embodiment, the dual agonist is administered to a patient at a dose of 1.5 mg to 7.5 mg. In one embodiment, the dual agonist is administered to a patient at a dose of 1.5 mg to 7.0 mg, 1.5 mg to 6.0 mg, 1.5 mg to 5.0 mg, 1.5 mg to 4.0 mg, or 1.5 mg to 3.5 mg. In one embodiment, the dual agonist is administered to a patient at a dose of 2.0 mg to 7.5 mg, 2.0 mg to 7.0 mg, 2.0 mg to 6.0 mg, 2.0 mg to 5.0 mg, 2.0 mg to 4.0 mg, or 2.0 mg to 3.5 mg. In one embodiment, the dual agonist is administered to a patient at a dose of 2.25 mg to 7.5 mg, 2.25 mg to 7.0 mg, 2.25 mg to 6.0 mg, 2.25 mg to 5.0 mg, 2.25 mg to 4.0 mg, or 2.25 mg to 3.5 mg. In one embodiment, the dual agonist is administered to a patient at a dose of 4.0 mg to 7.5 mg. In one embodiment, the dual agonist is administered to a patient at a dose of 4.0 mg to 6.0 mg.

[0114] In one embodiment, the dose is greater than 0.6 mg. In one embodiment, the dual agonist is administered to the patient at a dose of 1.5 mg.

[0115] In some embodiments, the dual agonist is administered to a patient at a dose of about 1.0 mg, about 1.5 mg, about 2.0 mg, about 2.25 mg, about 2.5 mg, about 3.0 mg, about 3.5 mg, about 4.0 mg, about 4.5 mg, about 5.0 mg, about 5.5 mg, about 6.0 mg, about 6.5 mg, about 7.0 mg, about 7.5 mg, about 8.0 mg, about 9.0 mg, or about 10.0 mg. In some embodiments, the dual agonist is administered to a patient at a dose of 1.0 mg, 1.5 mg, 2.0 mg, 2.25 mg, 2.5 mg, 3.0 mg, 3.5 mg, 4.0 mg, 4.5 mg, 5.0 mg, 5.5 mg, 6.0 mg, 6.5 mg, 7.0 mg, 7.5 mg, 8.0 mg, 9.0 mg, or 10.0 mg.

[0116] Administration Administration of the dual agonists described herein can be by any mode of administration common or standard in the art, for example, oral, intravenous, intramuscular, subcutaneous, sublingual, intranasal, intradermal, suppository, or implant. In a preferred embodiment of the invention described herein, administration is by subcutaneous injection.

[0117] The dosing regimens of the present invention can involve administering two or more doses of the dual agonist. Accordingly, in some embodiments, the present invention provides a GLP-1 / GLP-2 dual agonist or a pharmaceutically acceptable salt or solvate thereof described herein for use in a method for reducing or inhibiting weight gain, reducing food intake, reducing appetite, promoting weight loss, or treating obesity, morbid obesity, obesity-related gallbladder disease, or obesity-induced sleep apnea, comprising one or more administrations of the dual agonist to a patient at a dose of about 0.1 mg to 10.0 mg. In some embodiments, the method comprises two or more administrations of the dual agonist at a dose of about 0.1 mg to 10.0 mg to a patient. In some embodiments, each administration of the dual agonist to a patient is a dose of about 0.1 mg to 10.0 mg.

[0118] In some embodiments of the present invention, where the method comprises administering a dual agonist to a patient two or more times, the dose of the dual agonist may be different in each administration. In other words, the dose of the dual agonist need not be the same in each administration. However, in other embodiments of the present invention, where the method comprises administering a dual agonist to a patient two or more times, the dose of the dual agonist may be the same or substantially the same in each administration.

[0119] In some embodiments of the present invention, a series of single doses are delivered to a patient, and the dose of the dual agonist in the single-dose formulation may be increased after the first single-dose course. In some embodiments, the first course may include any one of 2, 3, 4, 5, 6, 7, 8, 9, 10, or more doses in which the amount of dual agonist in the single-dose formulation increases. In some embodiments, the dose of the dual agonist in the single-dose formulation after the first course may be the same as the last dose of the first course, less than the last dose of the first course, or more than the last dose of the first course. In certain embodiments, the dose of the dual agonist in the single-dose formulation after the first course may be the same as or approximately the same as the last dose of the first course.

[0120] The administration may involve weekly administration of the dual agonist. The term "weekly" refers to approximately every 7 days, e.g., approximately every 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, or 9 days, with each "day" counting as approximately 24 hours. As is understood in the art, the time between doses may vary somewhat, so not all doses are separated by exactly the same amount of time. This is often prescribed at the discretion of the physician. Thus, the doses may be separated by a clinically acceptable time range. In one embodiment of the invention described herein, the term "weekly" may refer to 7 days ± 2 days. That is, administration may occur up to 2 days before or up to 2 days after the stated date. Thus, administration may occur 2 days or 1 day before, or 1 day or 2 days after the stated date. In one embodiment, the dual agonist is administered weekly at a dose of about 1.5 mg to about 10.0 mg, e.g., about 1.5 mg to about 7.5 mg, e.g., about 1.5 mg to about 6.0 mg, e.g., about 1.5 mg to about 4.0 mg, e.g., about 1.5 mg to about 3.5 mg. In one embodiment, the dual agonist is administered weekly at a dose of about 2.0 mg to about 7.5 mg, e.g., about 2.0 mg to about 6.0 mg, e.g., about 2.0 mg to about 4.0 mg, e.g., about 2.0 mg to about 3.5 mg. In one embodiment, the dual agonist is administered weekly at a dose of about 2.25 mg to about 3.5 mg. In one embodiment, the dual agonist is administered weekly at a dose of 1.5 mg to 10.0 mg, e.g., about 1.5 mg to about 7.5 mg, e.g., 1.5 mg to 6.0 mg, e.g., 1.5 mg to 4.0 mg, e.g., 1.5 mg to 3.5 mg. In one embodiment, the dual agonist is administered weekly at a dose of 2.0 mg to 7.5 mg, e.g., 2.0 mg to 6.0 mg, e.g., 2.0 mg to 4.0 mg, e.g., 2.0 mg to 3.5 mg. In one embodiment, the dual agonist is administered weekly at a dose of 2.25 mg to 3.5 mg.

[0121] In one embodiment, the number of doses administered to the patient can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 or more doses. In other words, in some embodiments, the dual agonist is administered to the patient 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 or more times. In some embodiments, the methods comprise administering the dual agonist 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 or more times. In some embodiments, the dual agonist is administered to the patient at a dose of about 0.1-10.0 mg (or any other dose described herein) 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 or more times. In some embodiments, the method comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 or more administrations of the dual agonist at a dose of about 0.1-10.0 mg (or at any other dose described herein). In one embodiment, 4 doses are administered to the patient. In one embodiment, the method comprises 4 administrations of the dual agonist at a dose of about 0.1-10.0 mg (or at any other dose described herein). In one embodiment, 12 doses are administered to the patient. In one embodiment, the method comprises 12 administrations of the dual agonist at a dose of about 0.1-10.0 mg (or at any other dose described herein).

[0122] In one embodiment, the agonist can be administered in the same dose each time. In one embodiment, each administration of the dual agonist to the patient is in a dose of about 0.1 mg to 10.0 mg.

[0123] In one embodiment, multiple doses are administered to the patient over a period of weeks or months, or for a year or more.

[0124] In one embodiment, multiple doses are administered to the patient weekly and over a period of several weeks or months, or for a year or more.

[0125] In one aspect, the agonist may be administered in escalating doses.

[0126] Titration and Treatment The dose of the dual agonist according to the present invention may be a titration dose or a therapeutic dose.

[0127] The term "titration dose" refers to the dose of dual agonist administered to a patient at each administration during a titration period prior to administration at a therapeutic dose. Each titration dose is an amount of dual agonist between 0.1 mg and 10.0 mg. The doses, dosing regimes, and administration protocols presented herein apply equally to titration doses.

[0128] The term "therapeutic dose" refers to the dose of dual agonist administered to a patient at each administration during a treatment period. Each therapeutic dose is an amount of dual agonist between 0.1 mg and 10.0 mg. The doses, dosing regimes, and administration protocols presented herein apply equally to therapeutic doses.

[0129] In some embodiments, the dual agonist is administered to the patient according to a titration regimen, which includes an initial set of one or more administrations of the dual agonist during a "titration period," followed by a set of one or more administrations of the dual agonist during a "treatment period." Typically, the dose of the dual agonist in each administration during the titration period is lower than the dose in each administration during the treatment period.

[0130] The primary purpose of the titration period is to familiarize the patient with the side effects of the dual agonist. The initial administration of the dual agonist may cause side effects, which decrease in severity after further administration as the patient adapts. Administering a lower dose of the dual agonist during the titration period may reduce the initial severity of these side effects. The secondary purpose of the titration period is to determine the appropriate dual agonist dose for the patient. The dose of the dual agonist can be increased throughout the titration period, allowing the physician to observe side effects at different doses and thus determine the appropriate dose for treatment.

[0131] Thus, in some embodiments, the present invention provides a GLP-1 / GLP-2 dual agonist, or a pharmaceutically acceptable salt or solvate thereof, as described herein for use in a method for reducing or inhibiting weight gain, reducing food intake, reducing appetite, promoting weight loss, or treating obesity, morbid obesity, obesity-related gallbladder disease, or obesity-induced sleep apnea, comprising at least one administration of the dual agonist to a patient at a dose of about 0.1 mg to 10.0 mg, and comprising at least one administration of a titrating dose of the dual agonist to the patient and at least one administration of a therapeutic dose of the dual agonist to the patient. In other words, in some embodiments, the method comprises administering the dual agonist to the patient at least once at a titrating dose and at least once at a therapeutic dose.

[0132] In some embodiments, the method comprises administering to the patient two or more titrated doses of the dual agonist (i.e., two or three or more administrations). In some embodiments, the method comprises administering to the patient three or four or more, four or five or more, or five or six or more titrated doses of the dual agonist. In some embodiments, the method comprises administering to the patient one, two, three, four, or five titrated doses of the dual agonist. In preferred embodiments, the method comprises administering to the patient two titrated doses of the dual agonist. In preferred embodiments, the method comprises administering to the patient five titrated doses of the dual agonist.

[0133] In one embodiment, there may be at least one initial titration period of low doses before increasing the dose. In one embodiment, the titration period may consist of 1, 2, 3, or 4 low doses, where the dose is preferably the same each time. In one embodiment, the titration period consists of 1 low dose. In one embodiment, the titration period consists of 2 low doses.

[0134] In preferred embodiments, the titrated dose is administered weekly. In other words, in some embodiments, the method comprises administering a titrated dose of a dual agonist to a patient once a week.

[0135] The titration dose can be any dual agonist dose described elsewhere herein. In some embodiments, the titration dose is about 0.1 mg to about 10.0 mg. In some embodiments, the titration dose is about 1.0 mg to about 6.0 mg, e.g., about 1.5 mg to about 6.0 mg. Thus, in some embodiments, the method comprises administering at least one titration dose of about 1.5 mg to about 6.0 mg of the dual agonist to the patient. In one embodiment, the titration dose is about 1.0 mg to about 4.0 mg, e.g., about 1.5 mg to about 4.0 mg. In one embodiment, the titration dose is about 1.0 mg to about 3.5 mg, e.g., about 1.5 mg to about 3.5 mg, or about 1.5 mg to about 3.0 mg. In one embodiment, the titration dose is 1.0 mg, 2.0 mg, 2.25 mg, 3.0 mg, 3.5 mg, 4.0 mg, 4.5 mg, 5.0 mg, 5.5 mg, or 6.0 mg, or is about 1.0 mg, about 2.0 mg, about 2.25 mg, about 3.0 mg, about 3.5 mg, about 4.0 mg, about 4.5 mg, about 5.0 mg, about 5.5 mg, or about 6.0 mg. In some embodiments, the titration dose is 2.0 mg. In some embodiments, the titration dose is 2.0 mg and is administered once a week. In some embodiments, the titration dose is 4.0 mg. In some embodiments, the titration dose is 4.0 mg and is administered once a week.

[0136] The titrated dose need not be the same with each administration. In other words, different titrated doses can be administered to a patient within a titration period. Thus, in some aspects, the present invention provides a GLP-1 / GLP-2 dual agonist, or a pharmaceutically acceptable salt or solvate thereof, as described herein for use in a method of reducing or inhibiting weight gain, reducing food intake, reducing appetite, promoting weight loss, or treating obesity, morbid obesity, obesity-related gallbladder disease, or obesity-induced sleep apnea, comprising at least one administration of about 0.1 mg to 10.0 mg of the dual agonist to a patient, and comprising at least one administration of one or more titrated doses of the dual agonist to the patient and at least one administration of a therapeutic dose of the dual agonist to the patient.

[0137] In some embodiments, the method includes two or more, three or more, or four or more, different titrated doses. In some embodiments, the method includes two, three, or four different titrated doses. In preferred embodiments, the method includes two different titrated doses. Each titrated dose can be any of the dual agonist doses described elsewhere herein.

[0138] In some embodiments, all titration doses are the same (ie, there is one titration dose that is common to all administrations of the dual agonist to the patient during the titration period).

[0139] In some embodiments, the method comprises administering to the patient one dose of a 3.5 mg titrated dose of the dual agonist. In some embodiments, the method comprises administering to the patient two doses of a 2.0 mg titrated dose of the dual agonist. In some embodiments, the method comprises administering to the patient two doses of a 2.0 mg titrated dose of the dual agonist and three doses of a 4.0 mg titrated dose of the dual agonist.

[0140] In some embodiments, the methods involve administering to the patient two or more therapeutic doses of the dual agonist (i.e., two or three or more administrations). In some embodiments, the methods involve administering to the patient three or four or more, four or five or more, five or six or more, six or seven or more, seven or eight or more, eight or nine or more, nine or ten or more, ten or eleven or more, eleven or twelve or more, or twelve or thirteen or more therapeutic doses of the dual agonist. In some embodiments, the methods involve administering to the patient 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 therapeutic doses of the dual agonist. In preferred embodiments, the methods involve administering to the patient three therapeutic doses of the dual agonist. In preferred embodiments, the methods involve administering to the patient ten therapeutic doses of the dual agonist. In a preferred embodiment, the method comprises administering to the patient seven therapeutic doses of a dual agonist.

[0141] The therapeutic dose may continue to be administered for as long as necessary. The therapeutic dose of the dual agonist may be administered to a patient for a period of, for example, 1 month to 20 years, such as 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, 12 years, 13 years, 14 years, 15 years, 16 years, 17 years, 18 years, 19 years, or 20 years.

[0142] For example, a therapeutic dose can be administered weekly to a patient over a period of, for example, 1 month to 20 years, e.g., 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, 12 years, 13 years, 14 years, 15 years, 16 years, 17 years, 18 years, 19 years, or 20 years.

[0143] The therapeutic dose can be any dual agonist dose described herein. In some embodiments, the therapeutic dose is about 0.1 mg to about 10.0 mg. In some embodiments, the therapeutic dose is about 1.0 mg to about 10.0 mg, about 1.5 mg to about 10.0 mg, about 2.0 mg to about 10.0 mg, about 2.25 mg to about 10.0 mg, about 3.0 mg to about 10.0 mg, about 4.0 mg to about 10.0 mg, about 5.0 mg to about 10.0 mg, about 6.0 mg to about 10.0 mg, about 7.0 mg to about 10.0 mg, about 8.0 mg to about 10.0 mg, or about 9.0 mg to about 10.0 mg. In some embodiments, the therapeutic dose is about 1.0 mg, about 1.5 mg, about 2.0 mg, about 2.25 mg, about 2.5 mg, about 3.0 mg, about 3.5 mg, about 4.0 mg, about 4.5 mg, about 5.0 mg, about 5.5 mg, about 6.0 mg, about 6.5 mg, about 7.0 mg, about 7.5 mg, about 8.0 mg, about 9.0 mg, or about 10.0 mg. In some embodiments, the therapeutic dose is 1.0 mg, 1.5 mg, 2.0 mg, 2.25 mg, 2.5 mg, 3.0 mg, 3.5 mg, 4.0 mg, 4.5 mg, 5.0 mg, 5.5 mg, 6.0 mg, 6.5 mg, 7.0 mg, 7.5 mg, 8.0 mg, 9.0 mg, or 10.0 mg.

[0144] Typically, in embodiments where a dual agonist is administered to a patient more than once in a therapeutic dose (i.e., multiple administrations of a therapeutic dose of the dual agonist), all administrations during the treatment period are the same dose. Thus, in some embodiments, all therapeutic doses are the same (i.e., there is one therapeutic dose that is common to all administrations of the dual agonist to a patient during the treatment period).

[0145] However, therapeutic dose does not have to be the same in each administration.In other words, different therapeutic doses can be administered to patient during treatment period.The therapeutic dose can be changed according to the patient's response to dual agonist.For example, if patient shows severe side effects at a given therapeutic dose, the therapeutic dose can be reduced in subsequent administrations to reduce the severity of side effects.

[0146] Accordingly, in some aspects, the present invention provides a GLP-1 / GLP-2 dual agonist, or a pharmaceutically acceptable salt or solvate thereof, as described herein, for use in a method of reducing or inhibiting weight gain, reducing food intake, reducing appetite, promoting weight loss, or treating obesity, morbid obesity, obesity-related gallbladder disease, or obesity-induced sleep apnea, comprising at least one administration of the dual agonist to a patient at a dose of about 0.1 mg to 10.0 mg, and comprising at least one administration of one or more titration doses of the dual agonist to the patient, and at least one administration of one or more therapeutic doses of the dual agonist to the patient.

[0147] In some embodiments, the methods include two or more, three or more, or four or five or more different therapeutic doses. In some embodiments, the methods include two, three, or four different therapeutic doses. Each therapeutic dose can be any of the dual agonist doses described elsewhere herein.

[0148] Typically, a therapeutic dose is greater than a titration dose. Thus, in some aspects, a therapeutic dose is greater than a titration dose. In some aspects, a therapeutic dose is greater than some or all of the titration doses.

[0149] However, the therapeutic dose may be less than the titrated dose. For example, this may be the case when the titrated dose increases as the titration period progresses (i.e., the titrated dose increases over successive administrations), but the therapeutic dose is reduced as the titrated dose increases to account for side effects experienced by the patient. Thus, in some embodiments, the therapeutic dose is less than the titrated dose. In some embodiments, the therapeutic dose is less than some or all of the titrated dose.

[0150] As described herein, the purpose of dose titration is to identify an appropriate therapeutic dose. Thus, in some embodiments, the therapeutic dose is determined by a physician observing the effect of the titrated dose on the patient. In other words, the therapeutic dose may depend on the titrated dose.

[0151] In preferred embodiments, the therapeutic dose is administered weekly.In other words, in some embodiments, the method comprises administering to the patient a therapeutic dose of the dual agonist once a week.In some embodiments, the method comprises administering to the patient a titration dose of the dual agonist once a week and a therapeutic dose once a week.In other words, the weekly administration of the therapeutic dose of the dual agonist is a continuation of the weekly administration of the titration dose.

[0152] In one embodiment, the titration period may be followed by one or more doses at a dose greater than the titration dose. In one embodiment, the titration period is followed by one, two, three, or four doses at a dose greater than the titration dose. In one embodiment, the titration period is followed by three doses at a dose greater than the titration dose. In one embodiment, the titration period is followed by ten doses at a dose greater than the titration dose. In one embodiment, the titration period is followed by seven doses at a dose greater than the titration dose. In one embodiment, the titration period consists of one dose, followed by three doses at a dose greater than the titration dose. In one embodiment, the titration period consists of two doses, followed by ten doses at a dose greater than the titration dose. In one embodiment, the high dose is between about 3 mg and about 8 mg. In one embodiment, the high dose is between about 3 mg and about 8 mg.

[0153] In some embodiments, the method comprises administering to the patient one titration dose of 3.5 mg of the dual agonist, and administering to the patient three therapeutic doses of 6.0 mg of the dual agonist, each administration once per week.

[0154] In some embodiments, the method comprises administering to the patient two titration doses of 2.0 mg of the dual agonist and ten therapeutic doses of 4.0 mg of the dual agonist, each administration once a week.

[0155] In some embodiments, the method includes administering to the patient two titration doses of a 2.0 mg dual agonist, three titration doses of a 4.0 mg dual agonist to the patient, and seven therapeutic doses of a 6.0 mg dual agonist to the patient, each administration once weekly.

[0156] In one embodiment, the high dose (after a titration period) is at or about 6.0 mg, 7.0 mg, 7.5 mg, or 8.0 mg, preferably 6.0 mg.

[0157] Titrated doses may be administered daily, every other day, or weekly.

[0158] Post-titration doses are administered daily, every other day, or weekly.

[0159] Advantageously, subjects do not experience nausea or vomiting (or other adverse gastrointestinal effects) during the titration period, which allows for a shorter or more rapid titration period before administering a higher dose.

[0160] In one embodiment, there may be more than one titration period.

[0161] In one aspect of the invention, further doses are administered after the doses discussed above, i.e., the subject may continue to receive medication after the initial dose discussed herein.

[0162] Additional dosing may be once a week.

[0163] Administration of the dual agonist may continue for as long as necessary.

[0164] The additional doses can be administered as needed, for example, over a period of 1 month to 20 years, e.g., 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, 12 years, 13 years, 14 years, 15 years, 16 years, 17 years, 18 years, 19 years, or 20 years.

[0165] In a preferred embodiment, the dual agonist is administered to the subject daily.

[0166] medical conditions The dual agonists described herein have both GLP-1 and GLP-2 biological activity.

[0167] GLP-1 is a peptide hormone known for its important role in glucose homeostasis. When secreted from the gastrointestinal tract in response to nutrient intake, GLP-1 enhances glucose-stimulated insulin secretion from β-cells (Kim and Egan, 2008, Pharmacol. Rev. 470-512). Furthermore, GLP-1 or its analogs have been shown to increase somatostatin secretion and suppress glucagon secretion (Holst JJ, 2007, Physiol. Rev. 1409-1439).

[0168] In addition to its primary effect on glucose-stimulated insulin secretion, GLP-1 has been shown to be an important regulator of appetite, food intake, and body weight. Furthermore, GLP-1 can inhibit gastric emptying and gastrointestinal motility in both rodents and humans, most likely via GLP-1 receptors present in the gastrointestinal tract (Holst JJ, 2007, Physiol Rev. 1409-1439; Hellstrom et al., 2008, Neurogastroenterol Motil. Jun; 20(6):649-659). In addition, GLP-1 is thought to have insulin-like effects in major extrapancreatic tissues and is involved in glucose homeostasis and lipid metabolism in tissues such as muscle, liver, and adipose tissue (Kim and Egan, 2008, Pharmacol. Rev. 470-512).

[0169] In combination with an amylin analog, the dual agonist compounds described herein are also useful in, among other things, reducing or inhibiting weight gain, reducing the rate of gastric emptying or intestinal transit, reducing food intake, reducing appetite, or promoting weight loss. The effect on body weight may be mediated in part or in whole by reducing food intake, appetite, or intestinal transit.

[0170] Thus, the dual agonists of the present invention can be used to prevent or treat obesity, morbid obesity, obesity-related gallbladder disease, and obesity-induced sleep apnea.

[0171] Independent of their effect on body weight, the dual agonists of the present invention may have beneficial effects on glucose tolerance and / or glucose control. They may also be used to modulate (e.g., improve) circulating cholesterol levels, lower circulating triglyceride or LDL levels, and increase the HDL / LDL ratio.

[0172] They can therefore be used to prevent or treat poor glucose control, glucose tolerance or dyslipidemia (e.g., high LDL levels or low HDL / LDL ratio) and related conditions, including diabetes (e.g., type 2 diabetes, gestational diabetes), pre-diabetes, metabolic syndrome, and hypertension.

[0173] Many of these conditions are also associated with obesity or being overweight, and the effects of dual agonists on these conditions may therefore be due in whole or in part to their effect on body weight, or may be independent of their effect on body weight.

[0174] The effect on weight can be therapeutic or cosmetic.

[0175] The dual agonist activity of the compounds described herein can be particularly advantageous in many of the conditions described, as the two activities can complement each other.

[0176] For example, malabsorption is a condition resulting from abnormalities in the absorption of fluid and / or dietary nutrients, such as amino acids, sugars, fats, vitamins, or minerals, through the gastrointestinal (GI) tract, resulting in malnutrition and / or dehydration. Malabsorption can be the result of physical (e.g., traumatic) or chemical damage to the intestinal tract. The dual agonists described herein can improve intestinal barrier function, reduce gastric emptying, and increase intestinal absorption while simultaneously normalizing intestinal transit time. This not only helps patients increase nutrient and fluid absorption, but also reduces the patient's social concerns regarding food-induced bowel movements.

[0177] Furthermore, disorders of intestinal function and metabolism may be closely related to each other, with each contributing to the pathogenesis or symptomology of the other.

[0178] As noted above, obesity is associated with low-grade inflammation (sometimes described as "obesity-associated inflammation"). It is also generally recognized that obesity (along with other syndromes) causes increased vascular permeability, which allows pathogens and toxins such as LPS to penetrate the cell walls of the intestinal tract and cause inflammation. The changes that result from an inflammatory response are essentially the same, regardless of the cause and regardless of where the damage occurs. The inflammatory response can be acute (short-term) or chronic (long-lasting).

[0179] For example, obese mice (ob / ob and db / db mice) have been demonstrated to have disrupted mucosal barrier function and increased low-grade inflammation (Brun et al., 2007, Am. J. Physiol. Gastrointest. Liver Physiol., 292: G518-G525, Epub 5 Oct 2006). These findings were also observed in C57BL6 / J mice maintained on a high-fat diet (Cani et al., 2008, Diabetes, vol. 57, 1470-1481) and non-obese diabetic mice (Hadjiyanni et al., 2009, Endocrinology, 150(2): 592-599).

[0180] Cani et al. (Gut; 2009, 58:1091-1103) reported that modulation of the gut microbiota reduced intestinal barrier dysfunction and systemic inflammation via a GLP-2-dependent pathway in ob / ob mice. Furthermore, increased intestinal permeability observed in obese and diabetic patients likely plays a more important role in disease progression than previously anticipated. Increased intestinal permeability increases the transport of bacterial lipopolysaccharide (LPS) across the intestinal barrier. This increased LPS activates immune cells, including circulating macrophages and macrophages present in organs, resulting in low-grade chronic inflammation that may be involved in the pathogenesis of many diseases. This phenomenon is called metabolic endotoxemia (ME).

[0181] Inflammatory processes may also play a role in the development of metabolic dysfunction, such as insulin resistance and other metabolic disorders, in obese individuals.

[0182] Thus, the dual agonist compounds of the invention may be particularly useful in the prevention or treatment of low-grade inflammation, particularly in obese or overweight individuals, exerting beneficial effects through the GLP-1 agonist component of their activity and / or the GLP-2 component of their activity.

[0183] The therapeutic effectiveness of treatment with the dual agonists of the present invention can be monitored by taking intestinal biopsies to examine villous morphology, by biochemical assessment of nutrient absorption, by non-invasive determination of intestinal permeability, by weight gain in the patient, or by improvement of symptoms associated with these conditions.

[0184] Amylin analogs Amylin analogs (sometimes referred to as compounds or peptides) can be suitably prepared by standard synthetic methods. Thus, peptides can be synthesized, for example, by methods involving synthesis of peptides by standard solid-phase or solution-phase methodologies, either stepwise or by fragment assembly, and optional isolation and purification of the final peptide product. This method typically further includes the step of forming an amide bond between the side chains at positions 2 and 7, as described below. In the case of solid-phase synthesis, cyclization may be performed in situ on the solid phase (e.g., resin), i.e., before the peptide is removed from the solid phase.

[0185] Amylin analogs according to the present invention have the formula: R 1 -ZR 2 or a pharmaceutically acceptable salt or solvate thereof, wherein: R 1 is hydrogen, C 1-4 Acyl, benzoyl or C 1-4 alkyl, or a half-life extending moiety M, where M is optionally linked to Z via a linker moiety L; R 2 OH or NHR 3 where R 3 is hydrogen or C 1-3 alkyl, and Z is a group of formula I: X1-X2-X3-X4-X5-X6-X7-Ala-Thr-X10-Arg-Leu-Ala-X14-Phe-Leu-X17-Arg-X19-X20- Phe-Gly(Me)-Ala-Ile(Me)-X27-Ser-Ser-Thr-Glu-X32-Gly-Ser-X35-Thr-X37 (SEQ ID NO: 8) where: - X1 is selected from the group consisting of Arg, Lys, and Glu; - X3 is selected from the group consisting of Gly, Gln, and Pro; - X4 is selected from the group consisting of Thr and Glu; - X5 is selected from the group consisting of Ala and Leu; - X6 is selected from the group consisting of Thr and Ser; - X10 is selected from the group consisting of Glu and Gln; - X14 is selected from the group consisting of Aad, His, Asp, Asn, and Arg; - X17 is selected from the group consisting of Gln, His, and Thr; - X19-X20 are selected from Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly, and Ala-Asn, or are absent; - X27 is selected from the group consisting of Leu and Pro; - X32 is selected from the group consisting of Val and Thr; - X35 is selected from the group consisting of Asn and Ser; - X37 is selected from the group consisting of Hyp and Pro, and X2 and X7 are amino acid residues whose side chains together form a lactam bridge, - Gly(Me) is N-methylglycine (also known as sarcosine (Sar)); Ile(Me) is N-methylisoleucine, Aad is 2-aminoadipic acid, also known as homoglutamic acid, such as (2S)-2-aminoadipic acid [also known as (2S)-2-aminohexanedioic acid].

[0186] In a preferred embodiment, the amylin analog is [19CD]-isoGlu-RD()GTATK()ATERLA-Aad-FLQRSSF-Gly(Me)-A-Ile(Me)-LSSTEVGSNT-Hyp-NH2 (SEQ ID NO: 6) where: Gly(Me): N-methylglycine [also known as sarcosine (Sar)] Ile(Me): N-methylisoleucine Aad: 2-aminoadipic acid, also known as homoglutamic acid, e.g., (2S)-2-aminoadipic acid [also known as (2S)-2-aminohexanedioic acid]; Hyp: 4-hydroxyproline, e.g., (2S,4R)-4-hydroxyproline [also written as (4R)-4-hydroxy-L-proline] The backbone nitrogen of the [19CD]-isoGlu:Arg residue (present at position X1 of peptide sequence Z of the amylin analog) is covalently bound to the carboxyl group of the side chain of the Glu moiety via an amide bond, and the 19-carboxy-nonadecanoyl group is covalently bound to the alpha amino group of the Glu linker via an amide bond (as described below).

[0187] The parentheses "()" shown after the symbol of a particular amino acid residue indicate the residue whose side chain is involved in an intramolecular lactam bridge. Thus, the amylin analog compounds present in the formulations of the present invention have an intramolecular lactam bridge formed between the side chains of the residues shown in parentheses (aspartic acid and lysine, respectively).

[0188] The amylin analogue may be in the form of a pharmaceutically acceptable salt or solvate, for example a pharmaceutically acceptable acid addition salt.

[0189] Half-life extension moiety M As described herein, the N-terminal moiety R of the compounds of the present invention 1may be a half-life extending moiety M (sometimes described in the literature as, inter alia, a duration extending moiety or an albumin binding moiety), which may be linked (covalently bonded) to the peptide moiety Z via a linker moiety L. Suitable half-life extending moieties include certain types of lipophilic substituents. Without wishing to be bound by any particular theory, it is believed that such lipophilic substituents (and other classes of half-life extending moieties) bind to albumin in the bloodstream, thereby protecting the compounds of the invention from renal filtration and enzymatic degradation, and thus possibly extending the half-life of the compound in vivo. Lipophilic substituents may also modulate the potency of the compound as an agonist for the amylin (calcitonin) receptor.

[0190] The lipophilic substituent may be bonded to the N-terminal amino acid residue or the linker L via an ester, sulfonyl ester, thioester, amide, amine, or sulfonamide. It is therefore understood that the lipophilic substituent preferably comprises an acyl group, sulfonyl group, N atom, O atom, or S atom that forms part of an ester, sulfonyl ester, thioester, amide, amine, or sulfonamide. Preferably, the acyl group in the lipophilic substituent forms part of an amide or ester together with the amino acid residue or linker.

[0191] The lipophilic substituent may comprise a hydrocarbon chain having 10 to 24 carbon atoms, e.g., 14 to 22 carbon atoms, e.g., 16 to 20 carbon atoms. Preferably, the hydrocarbon chain has at least 14 carbon atoms, and preferably 20 or fewer carbon atoms. For example, the hydrocarbon chain may have 14, 15, 16, 17, 18, 19, or 20 carbon atoms. The hydrocarbon chain may be linear or branched, saturated or unsaturated. Furthermore, the hydrocarbon chain may contain a functional group at its terminus, e.g., a carboxylic acid group, which may or may not be protected during synthesis. From the above discussion, it will also be understood that the hydrocarbon chain is preferably substituted with a moiety that forms part of the bond to the N-terminal amino acid residue of the peptide moiety Z or to the linker L, e.g., an acyl group, a sulfonyl group, an N atom, an O atom, or an S atom.

[0192] Most preferably, the hydrocarbon chain is substituted with an acyl group, and therefore the hydrocarbon chain can be part of an alkanoyl group, such as dodecanoyl, 2-butyloctanoyl, tetradecanoyl, hexadecanoyl, heptadecanoyl, octadecanoyl, nonadecanoyl, or eicosanoyl. Examples of functionalized hydrocarbon chains are 15-carboxy-pentadecanoyl, 17-carboxy-heptadecanoyl, and 19-carboxy-nonadecanoyl.

[0193] As noted above, the lipophilic substituent M may be linked to the N-terminal amino acid residue of Z via a linker L. In embodiments, the linker moiety L may itself be linked to one, two, three or more linked sub-moieties L. 1 , L 2 , L 3 , ..., etc. When the linker L comprises only one such moiety, it is bonded to the lipophilic substituent and to the N-terminal amino acid residue of Z. The linker may then be bonded independently to the lipophilic substituent and to the N-terminal amino acid residue of Z by an ester bond, a sulfonyl ester bond, a thioester bond, an amide bond, an amine bond, or a sulfonamide bond. Thus, it may comprise two moieties independently selected from acyl, sulfonyl, an N atom, an O atom, and an S atom. The linker may be a linear or branched C 1-10 A hydrocarbon chain, or more preferably a linear C 1-5 The linker may be composed of a hydrocarbon chain. 1-6 Alkyl, Amino C 1-6 Alkyl, hydroxy C 1-6 Alkyl and carboxy C 1-6 It may be substituted with one or more substituents selected from alkyl.

[0194] In some embodiments, a linker can comprise one or more (e.g., one, two, or three) linked amino acid residues, which can each independently be the residue of any natural or unnatural amino acid. For example, a linker can comprise one, two, or three linked amino acid residues, each of which can independently be the residue Gly, Pro, Ala, Val, Leu, Ile, Met, Cys, Phe, Tyr, Trp, His, Lys, Arg, Gln, Asn, α-Glu, γ-Glu, ε-Lys, Asp, β-Asp, Ser, Thr, Gaba, Aib, β-Ala (i.e., 3-aminopropanoyl), 4-aminobutanoyl, 5-aminopentanoyl, 6-aminohexanoyl, 7-aminoheptanoyl, 8-aminooctanoyl, 9-aminononanoyl, 10-aminodecanoyl, or 8Ado (i.e., 8-amino-3,6-dioxaoctanoyl).

[0195] References to γ-Glu, ε-Lys, and β-Asp refer to amino acid residues that participate in the bond via their side chain carboxyl or amine functional groups. Thus, γ-Glu and β-Asp participate in the bond via their alpha amino group and side chain carboxyl group, while ε-Lys participates via its carboxyl group and side chain amino group. In the context of the present invention, γ-Glu and isoGlu are used interchangeably.

[0196] In certain embodiments, the linker comprises or consists of one, two, or three independently selected residues: Glu, γ-Glu, ε-Lys, β-Ala, 4-aminobutanoyl, 8-aminooctanoyl, or 8Ado.

[0197] Linkers composed of isoGlu and isoGlu-isoGlu may be particularly preferred.

[0198] An example of a lipophilic substituent comprising a lipophilic moiety M and a linker L is shown in the formula: [ka] In the formula, the backbone nitrogen of the Arg residue (located at position X1 in the peptide sequence Z of the amylin analog) is covalently linked to the carboxyl group of the side chain of the Glu moiety via an amide bond. The 19-carboxy-nonadecanoyl group is covalently linked to the alpha-amino group of the Glu linker via an amide bond. Thus, the Glu linker is in the iso-Glu (or γ-Glu) conformation. This combination of a lipophilic moiety and a linker attached to the Arg residue can be described, for example, by the shorthand notation [19CD]-isoGlu-R when shown in the formula of a particular compound.

[0199] Those skilled in the art will recognize suitable techniques for preparing compounds utilized in the context of the present invention. For examples of suitable chemistries, see, for example, WO 98 / 08871, WO 00 / 55184, WO 00 / 55119, Madsen et al. (J. Med. Chem. 2007, 50, 6126-32), and Knudsen et al., 2000 (J. Med. Chem. 43, 1664-1669).

[0200] The hydrocarbon chain in the lipophilic substituent may be further substituted. For example, it may be further substituted with up to three substituents selected from NH, OH, and COOH. When the hydrocarbon chain is further substituted, it is preferably further substituted with only one substituent. Alternatively, or in addition, the hydrocarbon chain may include, for example, a cycloalkane or heterocycloalkane moiety as shown below. [ka]

[0201] In some embodiments, the cycloalkane or heterocycloalkane moiety is a six-membered ring, for example, a piperidine ring.

[0202] In an alternative embodiment of the invention, the N-terminal amino acid of Z in the compounds of the invention may be linked (covalently bonded) to a biotinyl substituent, optionally via a linker moiety L. Without wishing to be bound by any particular theory, it is believed that such a biotinyl substituent similarly binds to albumin in the bloodstream, thereby protecting the compounds of the invention from enzymatic degradation and thus possibly extending the half-life of the compounds in vivo. The linker, if present, may provide space between the peptide moiety Z and the biotinyl substituent.

[0203] The biotinyl substituent may be attached to the N-terminal amino acid residue or to the linker via a maleimide ester bond, a sulfonyl ester bond, a thioester bond, an amide bond, an amine bond, or a sulfonamide bond. It will therefore be understood that the biotinyl substituent preferably comprises a maleimide group, an acyl group, a sulfonyl group, an N atom, an O atom, or an S atom that forms part of the ester bond, sulfonyl ester bond, thioester bond, amide bond, amine bond, or sulfonamide bond.

[0204] Examples of biotinyl substituents may include: [ka]

[0205] Biotin, also known as vitamin H or coenzyme R, is a water-soluble B vitamin (vitamin B7) that has been shown to increase the oral uptake of certain drugs.

[0206] Compound efficacy The compounds of the present invention are amylin receptor agonists, i.e., they are capable of binding to and inducing signal transduction by one or more receptors or receptor complexes believed to be physiological receptors for human amylin. These include the human calcitonin receptor hCT-R and complexes comprising the human calcitonin receptor hCT-R and at least one of the human receptor activity-modifying proteins designated hRAMP1, hRAMP2, and hRAMP3. The complexes between the hCT-R and hRAMP1, hRAMP2, and hRAMP3 are designated hAMYR1, hAMYR2, and hAMYR3 (i.e., human amylin receptors 1, 2, and 3), respectively.

[0207] A compound can be considered to be an amylin receptor agonist if it has agonist activity at one or more of hAMYR1, hAMYR2, and hAMYR3, eg, at hAMYR1 and / or hAMYR3, eg, at hAMYR3.

[0208] Typically, an amylin receptor agonist also has agonist activity at the hCT-R when expressed in the absence of hRAMP1, hRAMP2, and hRAMP3. Typically, an agonist has activity at the hCT-R (when expressed in the absence of hRAMP1, hRAMP2, and hRAMP3) in an equivalent assay that is less than 10-fold greater than its activity at any one of hAMYR1, hAMYR2, and hAMYR3 (i.e., its activity at all of these receptors). The agonist activity at hCT-R may be less than 5-fold greater than the agonist activity at hAMYR1, hAMYR2, and hAMYR3, may be substantially equal to (e.g., + / - 10%) the agonist activity at hAMYR1, hAMYR2, and hAMYR3, or may be less than the agonist activity at hAMYR1, hAMYR2, and hAMYR3. In this regard, it may be sufficient to compare the activity between hCT-R and hAMYR3.

[0209] The ability to induce cAMP formation (i.e., induce adenylate cyclase activity) as a result of binding to the relevant receptor or receptor complex is typically considered to be an indicator of agonist activity. Other intracellular signaling pathways or events can also be used as readouts for amylin receptor agonist activity. These can include calcium release, β-arrestin recruitment, receptor internalization, kinase activation or inactivation, lipase activation, inositol phosphate release, diacylglycerol release, or nuclear transcription factor translocation.

[0210] Suitable equivalent assay formats utilize cells that express hCT-R and differ only in their expression of hRAMP1, 2, and 3. For example, a "base" cell line that does not express hCT-R, hRAMP1, hRAMP2, or hRAMP3 can be engineered to generate cells that express (i) hCT-R and (ii) one of hAMYR1, hAMYR2, and hAMYR3 (i.e., hCT-R plus one of hRAMP1, hRAMP2, and hRAMP3), e.g., hAMYR3. Base cells are typically mammalian cells and may be primate cells. They may be non-human primate cells. Preferably, the base cells do not express any of CT-R, RAMP1, RAMP2, or RAMP3 (human, or native to the base cell if the base cell is non-human). The base cells may be fibroblasts. Suitable non-human fibroblast-based cells include African green monkey COS7 cells, which do not express native CT-R or RAMP.

[0211] Equivalent activity is determined by the EC 50 Determination of the value can be measured by any suitable means, for example via It will be apparent that the same biological readout must exist for both receptor types.

[0212] The compounds of the present invention may exhibit many advantageous properties relative to human amylin and its existing analogs, such as pramlintide, IAPP-GI, and analogs described in WO 2012 / 168430, WO 2012 / 168431, and WO 2012 / 168432. Compared to human amylin or any of these analogs, the compounds of the present invention may, for example, exhibit improved efficacy (e.g., in the form of improved in vitro activity or potency at one or more of the receptors hCT-R, hAMYR1, hAMYR2, or hAMYR3). Additionally, or alternatively, the compounds of the present invention may exhibit improved solubility in aqueous media, particularly at or across a range of pH values ​​from 4 to 7.5. Furthermore, the compounds of the present invention may also, or alternatively, exhibit a reduced tendency to undergo fibrillation in pharmaceutically relevant aqueous media, particularly at or across a range of pH values ​​from 4 to 7. Furthermore, the compounds of the present invention may also or alternatively exhibit improved chemical stability (i.e., reduced tendency to undergo chemical decomposition) in aqueous media at various pH values, particularly at or across the pH range of 4 to 9.

[0213] The compounds of the present invention are thus well suited for formulation in acidic media (e.g., pH 4) and neutral or near-neutral media (e.g., pH 7 or 7.4). In contrast to, for example, pramlintide, which typically exhibits low chemical stability and rapid fibrillation in pharmaceutically relevant aqueous media at neutral pH, the compounds of the present invention are thus well suited for co-formulation with, for example, insulin, various insulin analogs, and / or other therapeutic (e.g., antidiabetic or antiobesity) agents that require a neutral or near-neutral formulation pH.

[0214] Generally, as mentioned above, it is preferred to use the biological assay that measures the intracellular signal transduction caused by the binding of compound to relevant receptor.The activation of calcitonin / amylin receptor by the compound of the present invention (which acts as receptor agonist) induces the formation of cAMP and the activation of other intracellular signal transduction pathways and events.Therefore, the generation of cAMP or any other suitable parameter in the appropriate cell that expresses receptor can be used to monitor the agonist activity of receptor.

[0215] Those skilled in the art will be aware of suitable assay formats, and examples are provided below. For example, the assay can use the human calcitonin receptor (hCT-R, e.g., isoform 2 of the hCT-R) or the hAMYR3 receptor (see Examples below). Where reference is made to the sequence of a precursor protein, it should be understood that the assay can use the mature protein lacking the signal sequence.

[0216] EC 50 The EC value can be used as a numerical measure of agonist potency at a given receptor. 50 The EC value is a measure of the compound concentration required to achieve half of the compound's maximal activity in a particular assay. Thus, for example, the EC 50 Lower than [hCT-R] or EC 50 Lower EC than [hCT-R] 50 Compounds with [hCT-R] can be considered to have a potency or activity at the receptor that is greater than that of native human amylin or greater than that of pramlintide, respectively, in a particular assay.

[0217] In some embodiments of the compounds of the present invention, the EC 50 is less than 1.5 nM (for example, 0.001 to 1.5 nM).

[0218] In some embodiments of the compounds of the present invention, the EC50 is less than 0.9 nM (for example, 0.001 to 0.9 nM).

[0219] In some embodiments of the compounds of the present invention, the EC 50 is less than 0.5 nM (for example, 0.001 to 0.5 nM).

[0220] In some embodiments of the compounds of the present invention, the EC 50 is less than 0.3 nM (for example, 0.001 to 0.3 nM).

[0221] In some embodiments of the compounds of the present invention, the EC 50 is less than 0.2 nM (for example, 0.001 to 0.2 nM).

[0222] EC in hCT-R 50 may be an indicator of the effect of a compound on food intake, weight gain, and / or weight loss. EC 50 Compounds with low values ​​may have superior effects on these parameters.

[0223] In some embodiments of the compounds of the invention, the EC 50 is less than 1.0 nM (for example, 0.001 to 1.0 nM).

[0224] In some embodiments of the compounds of the invention, the EC 50 is less than 0.5 nM (for example, 0.001 to 0.5 nM).

[0225] In some embodiments of the compounds of the invention, the EC 50 is less than 0.4 nM (for example, 0.001 to 0.4 nM).

[0226] In some embodiments of the compounds of the invention, the EC 50 is less than 0.3 nM (for example, 0.001 to 0.3 nM).

[0227] In some embodiments of the compounds of the invention, the EC 50 is less than 0.2 nM (for example, 0.001 to 0.2 nM).

[0228] EC in hCT-R (expressed in the absence of hRAMP1, hRAMP2, and hRAMP3) 50 EC at any or all of hAMYR1, hAMYR2, and hAMYR3, e.g., at hAMYR3 50 It may be lower than that.

[0229] For example, EC in hCT-R (when expressed in the absence of hRAMP1, hRAMP2, and hRAMP3) 50 EC at any or all of hAMYR1, hAMYR2, and hAMYR3, e.g., at hAMYR3 50 It may be less than one-tenth of the

[0230] EC in hCT-R (expressed in the absence of hRAMP1, hRAMP2, and hRAMP3) 50 EC at any or all of hAMYR1, hAMYR2, and hAMYR3, e.g., at hAMYR3 50 It may be less than one-fifth of the

[0231] EC in hCT-R (expressed in the absence of hRAMP1, hRAMP2, and hRAMP3) 50 EC at any or all of hAMYR1, hAMYR2, and hAMYR3, e.g., at hAMYR3 50 may be substantially equal (e.g., + / - 50%) to

[0232] EC in hCT-R (expressed in the absence of hRAMP1, hRAMP2, and hRAMP3) 50 EC at any or all of hAMYR1, hAMYR2, and hAMYR3, e.g., at hAMYR3 50 It may be higher than that.

[0233] therapeutic use In combination with a dual agonist, amylin analogs are useful, among others, for reducing food intake, promoting weight loss, and inhibiting or reducing weight gain.They can therefore provide attractive treatment options for, among others, obesity and metabolic diseases caused by, characterized by, or associated with excess weight.Therefore, the compounds can be used in methods for treating, inhibiting, or reducing weight gain, promoting weight loss, reducing food intake, and / or reducing excess weight.Treatment can be achieved, for example, by controlling appetite, eating, food intake, calorie intake, and / or energy expenditure.

[0234] The compounds can be used in methods for treating obesity and related diseases, disorders, and conditions, including, but not limited to, morbid obesity, pre-surgical obesity, obesity-related inflammation, obesity-related gallbladder disease and obesity-induced sleep apnea and obesity-induced respiratory problems, cartilage degeneration, osteoarthritis, and reproductive health complications of obesity or being overweight, such as infertility.

[0235] The compounds may also be used in methods for preventing or treating Alzheimer's disease, diabetes, type 1 diabetes, type 2 diabetes, pre-diabetes, insulin resistance syndrome, impaired glucose tolerance (IGT), conditions associated with elevated blood glucose levels, metabolic diseases including metabolic syndrome, hyperglycemia, hypertension, atherogenic dyslipidemia, hepatic steatosis ("fatty liver", including non-alcoholic fatty liver disease (NAFLD), which itself includes non-alcoholic steatohepatitis (NASH)), renal failure, arteriosclerosis (e.g., atherosclerosis), macrovascular disease, microvascular disease, diabetic heart disease (including diabetic cardiomyopathy and heart failure, such as diabetic complications), coronary heart disease, peripheral arterial disease, or stroke.

[0236] The compounds may also be useful in lowering circulating LDL levels and / or increasing the HDL / LDL ratio.

[0237] The effects of the compounds may be mediated in whole or in part through an effect on body weight, or may be independent of an effect on body weight.

[0238] Metabolic syndrome is characterized by a group of metabolic risk factors in an individual, including abdominal obesity (excess adipose tissue surrounding internal organs), atherogenic dyslipidemia (a blood lipid disorder involving high triglycerides, low HDL cholesterol, and / or high LDL cholesterol, which contribute to the development of plaque deposits in arterial walls), elevated blood pressure (hypertension), insulin resistance and glucose intolerance, prothrombotic conditions (e.g., high fibrinogen or plasminogen activator inhibitor-1 in the blood), and inflammatory conditions (e.g., elevated C-reactive protein in the blood).

[0239] Individuals with metabolic syndrome are at increased risk for coronary heart disease and other diseases associated with other manifestations of atherosclerosis (e.g., stroke and peripheral vascular disease). The primary underlying risk factor for this syndrome is thought to be abdominal obesity.

[0240] Pharmaceutical Composition The invention also extends to compositions, e.g., pharmaceutical compositions, comprising amylin analogues. As with all aspects of the invention, reference to amylin analogues is understood to include reference to pharmaceutically acceptable salts and solvates.

[0241] The amylin analogs of the present invention can be formulated as pharmaceutical compositions suitable for administration, with or without preservation, which typically comprise a therapeutically effective amount of at least one peptide of the present invention together with a pharmaceutically acceptable carrier, excipient, or vehicle.

[0242] The term "pharmaceutically acceptable carrier" includes any standard pharmaceutical carrier. Pharmaceutically acceptable carriers for therapeutic use are well known in the pharmaceutical field and are described, for example, in "Remington's Pharmaceutical Sciences," 17th edition, Alfonso R. Gennaro (Ed.), Mark Publishing Company, Easton, PA, USA, 1985. For example, sterile saline and phosphate-buffered saline at slightly acidic or physiological pH can be used. Suitable pH buffers can be, for example, phosphate, citrate, acetate, tris(hydroxymethyl)aminomethane (TRIS), N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPS), ammonium bicarbonate, diethanolamine, histidine, arginine, lysine, or acetate (e.g., sodium acetate), or mixtures thereof. The term further encompasses all carrier agents listed in the United States Pharmacopoeia for use in animals, including humans.

[0243] The pharmaceutical compositions of the present invention may be in unit dosage form. In such form, the composition is divided into unit doses containing appropriate amounts of one or more active ingredients. The unit dosage form may be presented as a packaged preparation, the package containing discrete quantities of the preparation, for example, packaged tablets, capsules, or powders in vials or ampoules. The unit dosage form may also be, for example, a capsule, cachet, or tablet itself, or the appropriate number of any of these packaged forms. The unit dosage form may also be presented in single-dose injectable form, for example, in the form of a pen device containing a liquid phase (typically aqueous) composition. The composition may be formulated for any suitable route and means of administration. Pharmaceutically acceptable carriers or diluents include, for example, those used in formulations suitable for oral, intravitreal, rectal, vaginal, nasal, topical, enteral, or parenteral administration (including subcutaneous (sc), intramuscular (im), intravenous (iv), intradermal, and transdermal) administration, or by inhalation. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmaceutical formulation.

[0244] Subcutaneous or transdermal modes of administration may in some cases be suitable for the peptides of the present invention.

[0245] Further embodiments relate to devices, dosage forms, and packaging used to deliver the pharmaceutical formulations of the present invention. Thus, at least one peptide in a stable or preserved formulation or solution described herein can be administered to a patient in accordance with the present invention via a variety of delivery methods, including by sc or im injection, or by transdermal, pulmonary, or transmucosal administration, or by implant, or by use of an osmotic pump, cartridge, micropump, or other means recognized by one of skill in the art.

[0246] Further embodiments relate to oral formulations and oral administration. Formulations for oral administration may rely on the co-administration of adjuvants (e.g., resorcinol and / or nonionic surfactants, such as polyoxyethylene oleyl ether and n-hexadecyl polyethylene ether) to artificially increase the permeability of the intestinal wall and / or the co-administration of enzyme inhibitors (e.g., pancreatic trypsin inhibitor, diisopropylfluorophosphate (DFF), or trasylol) to inhibit enzymatic degradation. The active ingredient compound in a solid dosage form for oral administration may be mixed with at least one additive, such as sucrose, lactose, cellulose, mannitol, trehalose, raffinose, maltitol, dextran, starch, agar, alginate, chitin, chitosan, pectin, tragacanth gum, gum arabic, gelatin, collagen, casein, albumin, synthetic or semi-synthetic polymers, or glycerides. These dosage forms may also contain other types of additives, such as inert diluents, lubricants (such as magnesium stearate), parabens, preservatives (such as sorbic acid, ascorbic acid, or alpha-tocopherol), antioxidants (such as cysteine), disintegrants, binders, thickeners, buffers, pH adjusters, sweeteners, flavorings, or perfuming agents.

[0247] Dosage It will be understood that the dose of the GLP-1 / GLP-2 dual agonist and the dose of the amylin analog may be selected independently. The dose of the amylin analog may be selected as described herein.

[0248] Typical dosages of amylin analogs employed in the context of the present invention may range from about 0.0001 to about 100 mg per kg of body weight per day or every other day, e.g., about 0.0005 to about 50 mg per kg of body weight per day or every other day, e.g., about 0.001 to about 10 mg per kg of body weight per day or every other day, e.g., about 0.01 to about 1 mg per kg of body weight per day or every other day, administered in one or more doses, e.g., 1 to 3 doses. The exact dosage employed will depend, inter alia, on the nature and severity of the disease or disorder being treated, the sex, age, weight, and general condition of the subject being treated, any other possible concomitant diseases or disorders being or to be treated, and other factors known to a physician skilled in the art.

[0249] Typical dosages of amylin analogs employed in the context of the present invention may range from about 0.5 mg to about 10.0 mg per day or every other day, from about 0.6 mg to about 7.5 mg per day or every other day, preferably from about 1.2 mg to about 7.5 mg per day or every other day, preferably from about 1.2 mg to about 6.0 mg per day or every other day, preferably from about 2.4 mg to about 6.0 mg per day or every other day, preferably from about 2.4 mg to about 4.0 mg per day or every other day, preferably from about 2.4 mg to about 3.5 mg per day or every other day.

[0250] The amylin analogs of the present invention may be administered to a subject continuously (e.g., by intravenous administration or another continuous drug administration method) or at regular intervals, typically at regular time intervals, depending on the desired dosage for a particular subject and the pharmaceutical composition selected by one skilled in the art. Dosage intervals for regular administration include, for example, once daily, twice daily, every other day, every third day, every fourth day, every fifth day, or every sixth day, once or twice weekly, once or twice monthly, and the like. In certain circumstances, such as during chronic long-term administration, a regular peptide administration regimen may be advantageously interrupted for a period of time to reduce the level of the drug administered to the subject or to stop taking the drug; this is often referred to as taking a "drug holiday." Drug holidays are useful, for example, to maintain or regain sensitivity to a drug, particularly during intermittent long-term treatment, or to reduce undesirable side effects of long-term chronic treatment of a subject with a drug. The timing of the drug holiday depends on the timing of the regular dosing regimen and the purpose of taking the drug holiday (e.g., to regain sensitivity to the drug and / or to reduce undesirable side effects of continuous long-term administration). In some embodiments, the drug holiday can be a reduction in the dosage of the drug (e.g., to an amount below the therapeutically effective amount for a certain time interval). In other embodiments, administration of the drug is stopped for a certain time interval, after which administration begins again using the same or a different dosing regimen (e.g., a lower or higher dose and / or dosing frequency). The drug holiday of the present invention can therefore be selected from a wide range of durations and dosage regimens. Exemplary drug holidays are those of 2 or 3 days or more, 1 or 2 weeks or more, or 1 or 2 months or more, up to about 24 months. Thus, for example, a regular daily dosing regimen using a peptide of the invention can be interrupted by, for example, a one-week, two-week, or four-week rest period, after which the regular dosage regimen (e.g., a daily or weekly dosing regimen) is resumed. A variety of other rest period regimens are envisioned as useful for administering the peptides of the invention.

[0251] Thus, the peptide may be delivered via an administration regime comprising two or more administration steps separated by respective washout periods.

[0252] During each administration step, the peptide is administered to the recipient subject in a therapeutically effective amount according to a predetermined administration pattern. The administration pattern may include continuous administration of the drug to the recipient subject over the duration of the administration step. Alternatively, the administration pattern may include administration of multiple doses of the peptide to the recipient subject, the doses being separated by dosing intervals.

[0253] The dosing pattern may include at least 2 doses per dosing step, at least 5 doses per dosing step, at least 10 doses per dosing step, at least 20 doses per dosing step, at least 30 doses per dosing step, or more.

[0254] The dosing intervals can be regular dosing intervals, which can be as indicated above, including once daily, twice daily, once every 2, 3, 4, 5, or 6 days, once or twice weekly, once or twice monthly, or regular and less frequent dosing intervals, depending on the particular dosage formulation, bioavailability, and pharmacokinetic profile of the peptide.

[0255] The administration phase may have a duration of at least 2 days, at least 1 week, at least 2 weeks, at least 4 weeks, at least 1 month, at least 2 months, at least 3 months, at least 6 months, or more.

[0256] If the dosing pattern includes multiple doses, the duration of any possible subsequent drug holiday phases will be longer than the dosing intervals used in that dosing pattern. If the dosing intervals are irregular, the length of the drug holiday phase may be longer than the average interval between doses over the course of the dosing phase. Alternatively, the length of the drug holiday may be longer than the longest interval between successive doses during the dosing phase.

[0257] The duration of a possible drug holiday phase may be at least twice the duration of the relevant dosing interval (or its average), at least three times, at least four times, at least five times, at least ten times, or at least 20 times the duration of the relevant dosing interval or its average.

[0258] Within these constraints, a drug holiday phase may have a duration of at least 2 days, at least 1 week, at least 2 weeks, at least 4 weeks, at least 1 month, at least 2 months, at least 3 months, at least 6 months, or more, depending on the administration pattern during the previous administration phase.

[0259] A dosing regime involving the use of drug holidays comprises at least two dosing steps, with successive dosing steps separated by respective drug holiday steps. Thus, a dosing regime may comprise at least 3, at least 4, at least 5, at least 10, at least 15, at least 20, at least 25, or at least 30 or more dosing steps, each separated by a respective drug holiday step.

[0260] Successive administration phases may utilize the same administration pattern, although this is not always desirable or necessary. However, when other drugs or active agents are administered in combination with the peptides of the invention, the same combination of drugs or active agents is typically administered in the consecutive administration phases. In certain embodiments, the recipient subject is a human.

[0261] In a preferred embodiment, the amylin analog is administered to the subject every other day.

[0262] medical conditions The formulations of the present invention are useful, inter alia, in reducing food intake, promoting weight loss, and inhibiting or reducing weight gain, and may therefore provide attractive treatment options for, inter alia, obesity and metabolic disorders caused by, characterized by, or associated with excess body weight.

[0263] Thus, the formulations can be used in methods for treating, inhibiting, or reducing weight gain, promoting weight loss, reducing food intake, and / or reducing excess weight. Treatment can be achieved, for example, by controlling appetite, eating, food intake, calorie intake, and / or energy expenditure.

[0264] The formulations can be used in methods to treat obesity and related diseases, disorders, and conditions, including, but not limited to, morbid obesity, pre-surgical obesity, obesity-related inflammation, obesity-related gallbladder disease and obesity-induced sleep apnea and respiratory problems, cartilage degeneration, osteoarthritis, and reproductive health complications of obesity or being overweight, such as infertility.

[0265] The formulations can also be used in methods to prevent or treat Alzheimer's disease, diabetes, type 1 diabetes, type 2 diabetes, pre-diabetes, insulin resistance syndrome, impaired glucose tolerance (IGT), conditions associated with elevated blood glucose levels, metabolic diseases including metabolic syndrome, hyperglycemia, hypertension, atherogenic dyslipidemia, hepatic steatosis ("fatty liver", including non-alcoholic fatty liver disease (NAFLD), which itself includes non-alcoholic steatohepatitis (NASH)), renal failure, arteriosclerosis (e.g., atherosclerosis), macrovascular disease, microvascular disease, diabetic heart disease (including diabetic cardiomyopathy and heart failure, such as diabetic complications), coronary heart disease, peripheral arterial disease, or stroke.

[0266] The formulations may also be useful in lowering circulating LDL levels and / or increasing the HDL / LDL ratio.

[0267] These effects may be mediated in whole or in part through effects on body weight, or may be independent of effects on body weight.

[0268] Metabolic syndrome is characterized by a group of metabolic risk factors in an individual, including abdominal obesity (excess adipose tissue surrounding internal organs), atherogenic dyslipidemia (a blood lipid disorder involving high triglycerides, low HDL cholesterol, and / or high LDL cholesterol, which contribute to the development of plaque deposits in arterial walls), elevated blood pressure (hypertension), insulin resistance and glucose intolerance, prothrombotic conditions (e.g., high fibrinogen or plasminogen activator inhibitor-1 in the blood), and inflammatory conditions (e.g., elevated C-reactive protein in the blood).

[0269] Individuals with metabolic syndrome are at increased risk for coronary heart disease and other diseases associated with other manifestations of atherosclerosis (e.g., stroke and peripheral vascular disease). The primary underlying risk factor for this syndrome is thought to be abdominal obesity.

[0270] The term "treatment" (as well as "treating" and other grammatical variations thereof) as used in the context of the present invention refers to an approach for obtaining beneficial or desired clinical results. For purposes of this invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, attenuation of the extent of the disease, stabilization of the disease state (i.e., not worsening), delay or slowing of disease progression, improvement or reduction in pathology, and remission (partial or complete), whether detectable or undetectable. "Treatment" can also refer to prolonging survival as compared to expected survival in the absence of treatment. "Treatment" is an intervention performed with the goal of preventing the onset of a disorder or altering the pathology of a disorder. Thus, "treatment" refers to both therapeutic treatment and prophylactic or preventative treatment. When used in the context of prophylactic or preventative treatment, a pharmaceutical formulation need not completely prevent the onset of a disease or disorder. Subjects in need of treatment include those already suffering from the disorder and those in whom the onset of the disorder is being prevented. "Treatment" also means inhibiting or reducing the increase in pathology or symptoms (e.g., weight gain or hypoglycemia) compared to the absence of treatment, and does not necessarily include the complete cessation of the associated condition.

[0271] Devices and Kits In some embodiments, the present invention relates to a device for delivering the amylin analog or pharmaceutical composition of the present invention and / or the analog containing the dual agonist or pharmaceutical composition of the present invention to a subject. Via such a device, the dual agonist and / or amylin analog can be administered to a subject via a variety of delivery methods, including intravenous, subcutaneous, intramuscular, or intraperitoneal injection; oral administration; transdermal administration; pulmonary or transmucosal administration; administration by implant, osmotic pump, cartridge, or micropump; or other means recognized by those skilled in the art.

[0272] In some embodiments, the present invention relates to kits comprising the dual agonists and / or amylin analogs of the present invention or the pharmaceutical compositions of the present invention. In certain embodiments, the kits further comprise packaging and / or instructions for use.

[0273] Pharmaceutical Compositions and Administration One aspect of the present invention relates to separate compositions, wherein a first composition comprises a GLP-1 / GLP-2 dual agonist according to the present invention and / or a pharmaceutically acceptable salt or solvate thereof together with a carrier, and a second composition comprises an amylin analog according to the present invention and / or a pharmaceutically acceptable salt or solvate thereof together with a carrier. The first and second compositions may be prepared for storage or administration and comprise therapeutically effective amounts of a GLP-1 / GLP-2 dual agonist and, individually, an amylin analog of the present invention, or a salt or solvate thereof.

[0274] In one embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition in which the dual agonist is in the form of a pharmaceutically acceptable acid addition salt, and the pharmaceutical composition may include one or more pharmaceutically acceptable carriers, diluents, or excipients, and may also be referred to as a dual agonist pharmaceutical composition or dual agonist medicament.

[0275] In one embodiment, the pharmaceutical composition of the present invention is a pharmaceutical composition in which the amylin analog is in the form of a pharmaceutically acceptable acid addition salt, and the pharmaceutical composition may include one or more pharmaceutically acceptable carriers, diluents, or excipients, and may also be described as an amylin analog pharmaceutical composition or an amylin analog medicament.

[0276] In a preferred embodiment, the combination of the present invention comprises separate administration of an amylin analog pharmaceutical composition and an amylin analog pharmaceutical composition to a subject.

[0277] Therapeutic Uses, Methods, and Dosage Regimes In a broad aspect, the present invention provides one or more of a dual GLP-1 / GLP-2 agonist and an amylin analogue according to the invention for use as a medicament.

[0278] In one aspect, the present invention relates to a GLP-1 / GLP-2 dual agonist and an amylin analogue according to the present invention for use in therapy.

[0279] In one aspect, the present invention relates to a GLP-1 / GLP-2 dual agonist and an amylin analogue according to the present invention for use in the treatment or prevention of a disease or disorder.

[0280] In a preferred embodiment, the disease or disorder is obesity or obesity-related.

[0281] In one aspect, the present invention relates to a GLP-1 / GLP-2 dual agonist and an amylin analogue according to the present invention for use in a method of treating or preventing a disease or disorder.

[0282] In one aspect, the invention relates to a GLP-1 / GLP-2 dual agonist and an amylin analogue according to the invention for use in reducing the body weight of a subject.

[0283] The GLP-1 / GLP-2 dual agonists and amylin analogues according to the present invention can be used for therapeutic or cosmetic purposes.

[0284] The present invention further relates to a GLP-1 / GLP-2 dual agonist and an amylin analog for use in a method that is not itself for the treatment of a disease or disorder. Such uses and methods can be considered to be preventative of a disease or disorder insofar as they can be used before the onset of the disease (e.g., before diagnosis), for example, to improve an undesirable physiological characteristic or to alter a certain physiological parameter. Furthermore, such uses or methods can also be considered cosmetic.

[0285] It will be understood that the GLP-1 / GLP-2 dual agonists and amylin analogues for use in accordance with the present invention may be utilized in any of the diseases or disorders disclosed herein (e.g., in relation to each of the GLP-1 / GLP-2 dual agonists and amylin analogues).

[0286] It will be understood that the uses of the GLP-1 / GLP-2 dual agonists and amylin analogues of the invention described herein can be expressed as methods, and all features of the uses of the invention described herein are applicable to the corresponding methods of the invention.

[0287] Thus, the present invention provides methods for preventing or treating a disease or disorder comprising administering to a subject a GLP-1 / GLP-2 dual agonist and an amylin analog, hi some embodiments, the disease or disorder is obesity, morbid obesity, obesity-related gallbladder disease, obesity-induced sleep apnea, poor glucose control, glucose tolerance, dyslipidemia, diabetes, pre-diabetes, metabolic syndrome, or hypertension.

[0288] The present invention also provides methods for reducing or inhibiting weight gain, reducing food intake, reducing appetite, or promoting weight loss, comprising administering to a subject a GLP-1 / GLP-2 dual agonist and an amylin analog. The methods can be therapeutic or non-therapeutic (i.e., cosmetic).

[0289] It will likewise be understood that the uses of the GLP-1 / GLP-2 dual agonists and amylin analogues of the invention described herein may be expressed as dosing regimes, and all features of the uses of the invention described herein are applicable to the corresponding dosing regimes of the invention.

[0290] Thus, the present invention provides a dosing regimen for preventing or treating a disease or disorder comprising administering to a subject a GLP-1 / GLP-2 dual agonist and an amylin analog, hi some embodiments, the disease or disorder is obesity, morbid obesity, obesity-related gallbladder disease, obesity-induced sleep apnea, poor glucose control, glucose tolerance, dyslipidemia, diabetes, pre-diabetes, metabolic syndrome, or hypertension.

[0291] The present invention also provides a dosing regime for reducing or inhibiting weight gain, reducing food intake, reducing appetite, or promoting weight loss, comprising administering to a subject a GLP-1 / GLP-2 dual agonist and an amylin analog. The dosing regime can be therapeutic or non-therapeutic (i.e., cosmetic). [Example]

[0292] The following examples are provided to illustrate preferred embodiments of the invention and are not intended to limit the scope of the invention. [Example]

[0293] Effects of GLP-1 / GLP-2 dual agonists alone and in combination with amylin analogues in DIO rats The dual GLP-1 / GLP-2 agonist used in this example is the GLP-1 / GLP-2 dual agonist Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecanoyl]-isoGlu)]QAARDFIAWLIQHKITD-OH (SEQ ID NO: 5).

[0294] The amylin analog used in this example is amylin analog [19CD]-isoGlu-RD()GTATK()ATERLA-Aad-FLQRSSF-Gly(Me)-A-Ile(Me)-LSSTEVGSNT-Hyp-NH2 (SEQ ID NO: 6).

[0295] This study was conducted to evaluate the effects of treatment with a combination of a GLP-1 / GLP-2 dual agonist and an amylin analogue in diet-induced obese (DIO) rats.

[0296] Healthy male Sprague-Dawley rats were obtained from Taconic Denmark (Taconic Biosciences A / S). Rats weighed approximately 225–275 g upon arrival and were allowed to acclimate for 4 weeks. To generate diet-induced obese (DIO) rats for this study, animals were fed a high-fat diet (D12492, Research Diet Inc., New Brunswick, USA) ad libitum, with 60% of total energy derived from fat. Animals were housed in groups of two under a 12-h light / 12-h dark cycle (lights on 6:00 AM–6:00 PM) at standard temperature and humidity conditions (20–23°C, 50–80% relative humidity) and with free access to domestic-quality tap water.

[0297] DIO animals (n=9 / group) were stratified based on day 3 body weight and treated for 4 weeks, starting on day 0, with either vehicle (once daily (qd)), GLP-1 / GLP-2 dual agonist (100 nmol / kg, qd), amylin analog (10 nmol / kg, every other day (qod)), or a combination of GLP-1 / GLP-2 dual agonist and amylin analog (100 nmol / kg, qd and 10 nmol / kg, qod, respectively).

[0298] Outcomes included cumulative food intake, body weight (BW), and fasting blood glucose levels. Food intake was measured every other day from day -2 throughout the experiment, and cumulative food intake for each treatment group was calculated (Figure 1).

[0299] For BW outcomes, animals were weighed on day -2 (baseline weight) and daily throughout the study. The percent (%) change in BW from baseline was calculated for each day from days 0 to 28 (Figure 2A) and from baseline (day 0) to day 28 (the day before termination) (Figure 2B).

[0300] Overnight fasting blood glucose levels (mmol / L) were measured at baseline (day -1) and at termination (day 29) in overnight fasted animals. Blood samples (10 μl) were collected from the tail vein into heparin-coated capillary tubes using the Biosen enzyme-based electrode method with an S-line Biosen glucose analyzer (EKF diagnostics, US). The measured mean fasting blood glucose levels (mmol / L) for each group on day 29 and the mean blood glucose levels for all animals on day -1 (horizontal dotted lines) are shown in Figure 3. Exposure to both compounds was measured in plasma using LC-MS / MS.

[0301] Combination therapy with a GLP-1 / GLP-2 dual agonist and an amylin analog resulted in a significant, sustained, and greater cumulative reduction in food intake compared to vehicle (p<0.0001) and compared to GLP-1 / GLP-2 dual agonist monotherapy (p<0.05) (Figure 1), and also resulted in a significant, sustained, and greater reduction in BW compared to vehicle and either monotherapy treatment (Figures 2A and 2B). Data show changes compared to initial body weight of DIO animals of -12.4%±0.9 for GLP-1 / GLP-2 dual agonist, -5.9%±0.4 for amylin analog, -18.9%±1.8 for combination therapy, and +4.4%±0.7 for vehicle, with p<0.0001 for all groups compared to vehicle, p<0.01 for combination compared to GLP-1 / GLP-2 dual agonist, and p<0.0001 for combination compared to amylin analog.

[0302] GLP-1 / GLP-2 dual agonist monotherapy and combination treatments resulted in significant reductions in fasting blood glucose levels compared to vehicle, p<0.001 (FIG. 3).

[0303] Treatment with the GLP-1 / GLP-2 dual agonist, the amylin analog, and the combination of the two was observed to be well tolerated. Exposure of the GLP-1 / GLP-2 dual agonist and the amylin analog measured in plasma showed no difference between dosing as single compounds and when both compounds were administered together in a combination (Figure 4).

[0304] In conclusion, in the DIO rat model, a GLP-1 / GLP-2 dual agonist potentiates the effect of amylin analog monotherapy on weight loss, suggesting an additive effect of this combination therapy for the management of overweight, obesity, and obesity-related comorbidities.

[0305] All publications mentioned in the above specification are herein incorporated by reference. Various modifications and variations of the described methods and system of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention that are obvious to those skilled in biochemistry, molecular biology, or related fields are intended to be within the scope of the following embodiments.

Claims

1. 1. A GLP-1 / GLP-2 dual agonist and an amylin analogue for use in a method for preventing or treating a disease or disorder, wherein the GLP-1 / GLP-2 dual agonist and an amylin analogue are administered to a subject.

2. 2. The GLP-1 / GLP-2 dual agonist and amylin analogue for use according to claim 1, wherein the disease or disorder is obesity, morbid obesity, obesity-related gallbladder disease, obesity-induced sleep apnea, poor glucose control, glucose tolerance, dyslipidemia, diabetes, pre-diabetes, metabolic syndrome, or hypertension.

3. A GLP-1 / GLP-2 dual agonist and an amylin analogue for use in a method of reducing or inhibiting weight gain, reducing food intake, reducing appetite or promoting weight loss.

4. 4. The GLP-1 / GLP-2 dual agonist and amylin analogue for use according to any one of claims 1 to 3, wherein the GLP-1 / GLP-2 dual agonist is Hy-H[Aib]EGSFTSELATILD[K([17-carboxy-heptadecanoyl]-isoGlu)]QAARDFIAWLIQHKITD-OH (SEQ ID NO: 5), or a pharmaceutically acceptable salt or solvate thereof.

5. 5. The GLP-1 / GLP-2 dual agonist and amylin analogue for use according to any one of claims 1 to 4, wherein the method comprises administering to the subject a dose of the GLP-1 / GLP-2 dual agonist of from about 0.5 mg to about 10.0 mg.

6. 6. A GLP-1 / GLP-2 dual agonist and an amylin analogue for use according to any one of claims 1 to 5, wherein the method comprises administering to the subject the GLP-1 / GLP-2 dual agonist at a dose of about 0.5 mg to about 7.5 mg, preferably about 1.0 mg to about 7.5 mg, preferably about 1.0 to about 6.0 mg, preferably about 1.0 to about 4.0 mg, preferably about 1.0 to about 3.5 mg.

7. 7. The GLP-1 / GLP-2 dual agonist and amylin analogue for use according to any one of claims 1 to 6, wherein the method comprises administering the GLP-1 / GLP-2 dual agonist to the subject once daily at a dose of about 0.5 mg to about 10.0 mg per day.

8. 8. The GLP-1 / GLP-2 dual agonist and amylin analogue for use according to any of claims 1 to 7, wherein the method comprises administering to the subject the GLP-1 / GLP-2 dual agonist once daily at a dose of about 0.5 mg to about 7.5 mg per day, preferably once daily at a dose of about 1.0 mg to about 7.5 mg per day, preferably once daily at a dose of about 1.0 to about 6.0 mg per day, preferably once daily at a dose of about 1.0 to about 4.0 mg per day, preferably once daily at a dose of about 1.0 to about 3.5 mg per day.

9. The amylin analog is [19CD]-isoGlu-RD()GTATK()ATERLA-Aad-FLQRSSF-Gly(Me)-A-Ile(Me)-LSSTEVGSNT-Hyp-NH ２ (SEQ ID NO: 6), or a pharmaceutically acceptable salt or solvate thereof.

10. 10. A GLP-1 / GLP-2 dual agonist and amylin analogue for use according to any of claims 1 to 9, wherein the method comprises the step of administering to the subject the amylin analogue at a dose of about 0.5 mg to about 10.0 mg, preferably about 0.6 mg to about 7.5 mg, preferably about 1.2 mg to about 7.5 mg, preferably about 1.2 to about 6.0 mg, preferably about 2.4 to about 6.0 mg, preferably about 2.4 to about 4.0 mg, preferably about 2.4 to about 3.5 mg.

11. 11. A GLP-1 / GLP-2 dual agonist and amylin analogue for use according to any of claims 1 to 10, wherein the method comprises administering to the subject an amylin analogue at a dose of about 0.5 mg to about 10.0 mg per day every other day, preferably at a dose of about 0.6 mg to about 7.5 mg per day every other day, preferably at a dose of about 1.2 mg to about 7.5 mg per day every other day, preferably at a dose of about 1.2 to about 6.0 mg per day every other day, preferably at a dose of about 2.4 to about 6.0 mg per day every other day, preferably at a dose of about 2.4 to about 4.0 mg per day every other day, preferably at a dose of about 2.4 to about 3.5 mg per day every other day.

12. 12. The GLP-1 / GLP-2 dual agonist and amylin analogue for use according to any one of claims 1 to 11, wherein the GLP-1 / GLP-2 dual agonist and / or amylin analogue, or a pharmaceutically acceptable salt or solvate thereof, are independently mixed with a carrier, excipient, and / or vehicle.

13. 13. The GLP-1 / GLP-2 dual agonist and amylin analogue for use according to claim 12, wherein the carrier is a pharmaceutically acceptable carrier.

14. The amylin analog is in a pharmaceutical composition, said pharmaceutical composition comprising: (i) the one or more amylin analogs are at a concentration of about 0.4 mg / ml to about 25 mg / ml; (ii) being present in a solution having a buffer concentration of about 0.5 mM to 25 mM; (iii) in a solution having a pH of about 5.8 to about 6.9; and / or (iv) the amylin analog is provided as a chloride salt; 14. A GLP-1 / GLP-2 dual agonist and amylin analogue for use according to any one of claims 1 to 13, comprising one or more of:

15. The GLP-1 / GLP-2 dual agonist is in a pharmaceutical composition, the pharmaceutical composition comprising: (i) at least about 1 mg / mL of one or more GLP-1 / GLP-2 dual agonists (ii) 5 mM to about 50 mM of a phosphate buffer component (iii) about 190 mM to about 240 mM mannitol, and / or (iv) pH is about pH 8.0 15. A GLP-1 / GLP-2 dual agonist and amylin analogue for use according to any one of claims 1 to 14, comprising one or more of:

16. The GLP-1 / GLP-2 dual agonist and amylin analogue for use according to any one of claims 1 to 15, wherein the subject is a human.

17. 17. The GLP-1 / GLP-2 dual agonist and amylin analogue for use according to any of claims 1 to 16, wherein the GLP-1 / GLP-2 dual agonist and amylin analogue are administered by injection, preferably by subcutaneous injection.

18. A dosing regime for preventing or treating a disease or disorder comprising administering to a subject a GLP-1 / GLP-2 dual agonist and an amylin analogue.

19. A dosing regime for reducing or inhibiting weight gain, reducing food intake, reducing appetite, or promoting weight loss, comprising administering to a subject a GLP-1 / GLP-2 dual agonist and an amylin analog.

20. A dosing regime according to claim 18 or 19, comprising the features of any of claims 1 to 17.

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