Combination therapy comprising long-acting GLP-1 / glucagon and NPY2 receptor agonists

Abstract

The present invention provides a combination therapy comprising: a compound I (GLP-1R / glucagon receptor dual agonist): H-His-Ac4c-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Leu-Asp-Glu-Arg-Ala-Lys-Asp-Phe-Lys (HOOC-(CH2) 16-CO-gamma Glu-Gly-Ser-Gly-Gly-Gly-)-Trp-Leu-Glu-Ser-Ala-NH2 (I), and a long acting NPY2 receptor agonist selected from the group consisting of compounds A to L. The invention further relates to a method for preparing the same. The combination therapy may be used to treat obesity and various obesity-related conditions, diseases, or co-diseases such as type 2 diabetes, liver diseases such as NAFLD (non-alcoholic fatty liver disease) and NASH (non-alcoholic steatohepatitis), kidney diseases, or cardiovascular diseases.

Description

Technical Field

[0001] The present invention relates to the pharmacology and biology of a combination of long-acting acylated PYY analogs, which are neuropeptide Y2 (NPY2) receptor agonists, and dual glucagon-like peptide-1 (GLP-1) / glucagon (GCG) receptor agonists, and their medical uses for treating and / or preventing various diseases, disorders or conditions, such as treating and / or preventing excessive food intake, excess body weight, obesity, metabolic diseases and other disorders or conditions associated with excess body weight or obesity (e.g., diabetes), kidney disease, non-alcoholic steatohepatitis, and cardiovascular disease. Background Art

[0002] obesity

[0003] Obesity is a chronic, relapsing, progressive disease and one of the greatest healthcare challenges of our time. In 2016, more than 1.9 billion adults aged 18 years and over were overweight and more than 650 million of them were obese (BMI ≥ 30 kg / m 2 ). Despite long-term efforts, the number of overweight and obese people is still increasing and the prevalence of obesity almost tripled between 1975 and 2016. This translates to 39% of adults aged 18 years or older (39% of men and 40% of women) being overweight, and 13% being obese.

[0004] Overweight and obesity are defined as abnormal or excessive fat accumulation that poses a risk to health. In this regard, overweight and obesity are major risk factors for many chronic diseases, and the chronic diseases associated with direct (comorbidity) and indirect (complications) include, but are not limited to, cardiovascular disease (e.g., heart failure), cardiometabolic disease (such as insulin resistance), type 2 diabetes, atherosclerosis, cardiovascular disease, hypertension, dyslipidemia, hyperuricemia, chronic kidney disease, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), cirrhosis, depression, sleep apnea, musculoskeletal diseases (e.g., osteoarthritis), gallbladder disease, and certain types of cancer. In the United States, obesity is now believed to be the second leading cause of preventable death after smoking.

[0005] If current trends continue, it is projected that by 2025, 2.7 billion adults will be overweight, more than 1 billion will be affected by obesity, and 177 million adults will be severely affected by obesity. Rising obesity has led to an increase in diabetes, and approximately 90% of people with type 2 diabetes can be classified as obese. There are 246 million people with diabetes worldwide, and by 2025, it is projected that 380 million people will have diabetes.

[0006] First-line treatment for overweight and obese patients includes diet and exercise but is often not effective enough. Second-line treatment options are bariatric surgery and drug therapy. Available drug treatments appear to lack efficacy and / or safety, and only a limited number of approved therapies are available in the United States and Europe. Therefore, there remains a high medical need for more effective and safe treatment options.

[0007] Glucagon, GLP-1, and Oxytocin

[0008] Pre-proglucagon is a 158 amino acid precursor polypeptide that is differentially processed in tissues to form a number of structurally related proglucagon-derived peptides, including glucagon (Glu), glucagon-like peptide-1 (GLP-1), glucagon-like peptide-2 (GLP-2), and oxygenomodulin (OXM). These molecules are involved in a wide range of physiological functions, including glucose homeostasis, insulin secretion, gastric emptying and intestinal growth, and regulation of food intake. Glucagon is a 29 amino acid peptide that corresponds to amino acids 53 to 81 of pre-proglucagon. Oxytomodulin (OXM) is a 37 amino acid peptide that includes the complete 29 amino acid sequence of glucagon and an octapeptide carboxyl-terminal extension (amino acids 82 to 89 of pre-proglucagon). The major biologically active fragment of GLP-1 is produced as a 30 amino acid C-terminal amidated peptide that corresponds to amino acids 98 to 127 of pre-proglucagon. GLP-1 reduces high blood glucose concentrations by improving glucose-stimulated insulin secretion and promotes weight loss primarily by reducing food intake. Clinically, GLP-1 receptor agonists are established therapies for the treatment of diabetes and obesity, with a limited therapeutic window due to gastrointestinal side effects (such as nausea and vomiting) related to their mechanism. Glucagon helps maintain glucose concentrations in the blood by binding to glucagon receptors on hepatocytes, causing the liver to release glucose stored in the form of glycogen through glycogenolysis. When these reserves are exhausted, glucagon stimulates the liver to synthesize additional glucose through gluconeogenesis. Glucagon has been preclinically and clinically shown to affect body weight by increasing energy expenditure. This glucose is released into the blood, preventing the development of hypoglycemia. OXM is released into the blood in response to food intake and is proportional to the calorie content of the meal. OXM activates both glucagon and GLP-1 receptors, with OXM having a slightly higher potency for glucagon receptors than for GLP-1 receptors, but lower than the potency of native glucagon and GLP-1 for their respective receptors. Human glucagon is also able to activate both receptors, although with a stronger preference for the glucagon receptor over the GLP-1 receptor. GLP-1, on the other hand, is unable to activate the glucagon receptor. OXM is involved in regulating body weight and has been shown in humans to suppress appetite and inhibit food intake and energy expenditure.

[0009] Other peptides have been shown to bind to and activate both the glucagon receptor and the GLP-1 receptor and inhibit weight gain (see, e.g., WO2011 / 075393, WO2014 / 041195, WO2015 / 183054, WO2016 / 065090, WO2016 / 108617, WO2017 / 074798).

[0010] PYY

[0011] Peptide YY (PYY) is a 36-amino acid peptide secreted by endocrine cells (L cells) in the gastrointestinal tract in response to food intake and exhibits a feeding inhibitory effect via the Y2 receptor. 3-36 , N Engl J Med 2003;349;941-8). Enteric / hypothalamic pathways via Y2 receptors in neurons expressing NPY / AgRP in the hypothalamic arcuate nucleus, and vagal afferent pathways via Y2 receptors in vagal nerve terminals have been reported as anorectic mechanisms of action, which for native PYY(3-36) is associated with limited tolerability in humans due to dose-dependent nausea and vomiting when administered intranasally. PYY is cleaved to PYY(3-36) by dipeptidyl peptidase IV (DPP IV). Compared to PYY(1-36), PYY(3-36) shows selectivity for neuropeptide Y2 receptors over neuropeptide Y1, Y4 and Y5 receptors, although retaining some Y1 and Y5 affinity. However, PYY and PYY(3-36) have short half-lives in vivo and display undesirable chemical or physical properties, such as low stability. Furthermore, the pharmacological effects (eg, their utility as anti-obesity agents) appear to be limited, however, long-acting analogs of PYY3-36 have been shown preclinically and clinically to offer a therapeutic window that can achieve anti-obesity efficacy with acceptable tolerability.

[0012] WO2014 / 178018 discloses the ability of PYY analogs and the like to reduce food intake in mice.

[0013] WO2011 / 033068 and WO2011 / 058165 disclose long-acting NPY2R agonists.

[0014] WO2015 / 071355, WO2016 / 198682 and WO2020 / 092191 relate to PYY compounds which are selective NPY2R agonists. Disclosed herein are PYY compounds comprising covalently attached substituents or modifying groups, also referred to therein as prolonging moieties.

[0015] GLP-1 and NPY2 receptor agonism

[0016] Simultaneous activation of GLP-1R and NPY2R, and synergistic effects of reduced food intake and weight loss have been demonstrated in preclinical species, such as mice (Neary NM et al., Endocrinology 2005; 146: 5120; Talsania T et al., Endocrinology 2005; 146: 3748; Kjargaard M. et al., Neuropeptides 2019; 73: 89; WO2011 / 039096; WO2019 / 207505), rats (Reidelberger RD et al., Obesity 2011; 19: 121; Dischinger U et al., Frontiers in Endocrinology 2021; 11: 598843; WO2017 / 035432) and pigs (WO2015 / 071355).

[0017] In humans, administration (continuous infusion) of GLP-1 together with PYY3-36 (Schmidt JB et al., Am J Physiol Endocrinol Metab 2014; 306: E1248) or PYY3-36, GLP-1 and oxytocin (Field BCT et al., Diabetes 2010 59: 1635; Tan T et al., J. Clin. Endocrinol Metab 2017; 102: 2364; Tan T et al., Diabetes Care 2019, 42(8): 1446) demonstrated a synergistic effect in reducing energy intake.

[0018] The present invention provides a combination therapy comprising administration of a long-acting GLP-1R / GCGR dual agonist and a long-acting NPY2R agonist. The simultaneous activation of the GLP-1R, GCGR and NPY2 receptors has the potential to provide a very effective treatment method by reducing food intake, appetite and / or weight by simultaneously synergistically reducing energy intake through GLP-1R and NPY2R agonism and increasing energy expenditure through GCGR activation. According to the present invention, it is possible to activate the three receptors with a long-acting agent that allows administration by subcutaneous injection. In addition, the combination therapy can be effectively administered by once-weekly injection of the agonist.

[0019] The choice of NPY2R agonist to be combined with a GLP-1R / GCGR dual agonist appears to be important. However, the selection of an effective NPY2R agonist is not obvious in view of the data provided in the prior art. (e.g. WO2021 / 094259 or WO2022 / 029231).

[0020] Furthermore, the combination therapy according to the invention herein appears to provide a more sustained effect on food intake suppression when compared to a combination of a GLP-1R agonist and a NPY2R agonist or when compared to GLP-1R / GCGR alone. Summary of the invention

[0021] The present invention relates to a combination therapy comprising administering a long-acting GLP-1 / glucagon receptor dual agonist and a long-acting NPY2 receptor agonist.

[0022] More specifically, the present invention relates to a combination therapy comprising administering to a patient:

[0023] - An effective amount of Compound I:

[0024] H-His-Ac4c-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Arg-Ala-Ala-Lys-Asp-Phe-Ile-Lys(HOOC-(CH2) 16 -CO-γGlu-Gly-Ser-Gly-Ser-Gly-Gly-)-Trp-Leu-Glu-Ser-Ala-NH2 (SEQ ID NO: 01, SEQ ID NO 02);

[0025]

[0026] and

[0027] - A long-acting NPY2 receptor agonist selected from the group consisting of compounds A to L.

[0028] In one embodiment, the long-acting NPY2 receptor agonist is selected from the group consisting of Compounds B, E, K and L.

[0029] In one embodiment, the weight ratio of Compound 1 to NPY2 receptor agonist is in the range of 1:5 to 5:1 (Compound 1:NPY2 receptor agonist).

[0030] A PYY analogue selected from the group consisting of Compounds A to L is for use together with Compound I in combination therapy.

[0031] The compound may be administered once a week (once a week) or more frequently.

[0032] In one embodiment, Compound I and the long-acting NPY2 receptor agonist are administered via subcutaneous injection, for example, via two separate injections or via one injection where both compounds are administered simultaneously. In the case of two separate injections, one of the compounds may be administered first, followed immediately by the other, or there may be a period of time between the two injections, such as 4, 3, 2 or 1 day. Alternatively, the time between injections is 12, 6, 4, 2 or 1 hour. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 : Effect of Compound B on (A) food intake (grams) and (B) relative (percent) body weight in DIO mice. DIO animals were dosed once daily with different doses of Compound B. Relative body weight was measured daily as % of baseline body weight of each animal over a 28-day period. Food intake was measured in grams per animal per day over a 28-day period. Data are presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001; One-way ANOVA compared to vehicle group followed by Dunnett's multiple comparison test.

[0034] Figure 2 : Effect of Compound E on (A) food intake (grams) and (B) relative (percent) body weight in DIO mice. DIO animals were dosed once daily with different doses of Compound E. Relative body weight was measured daily as % of baseline body weight of each animal over a 28-day period. Food intake was measured in grams per animal per day over five 28-day periods. Data are presented as mean ± SEM. *p<0.001; one-way ANOVA compared to vehicle group followed by Dunnett's multiple comparison test.

[0035] Figure 3 : Effect of Compound K on (A) food intake (grams) and (B) relative (percent) body weight in DIO mice. DIO animals were dosed once daily with different doses of Compound K. Relative body weight was measured daily as % of baseline body weight of each animal over an 8-day period. Food intake was measured in grams per animal per day over an 8-day period. Data are presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001; one-way ANOVA compared to vehicle group followed by Dunnett's multiple comparison test.

[0036] Figure 4: Effect of Compound L on (A) food intake (grams) and (B) relative (percent) body weight in DIO mice. DIO animals were dosed once daily with different doses of Compound L. Relative body weight was measured daily as % of baseline body weight of each animal over an 8-day period. Food intake was measured in grams per animal per day over an 8-day period. Data are presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001; one-way ANOVA compared to vehicle group followed by Dunnett's multiple comparison test.

[0037] Figure 5 : Effects of Compound B alone or in combination with Semaglutide on (A) relative (percent) body weight, (B) food intake (grams), (C) cumulative food intake (grams), and (D) area under the curve of panel B in DIO mice. DIO animals were dosed once daily with different doses of Compound B alone or in combination with Semaglutide. Relative body weight was measured daily as % of baseline body weight of each animal over a 28-day period. Food intake was measured in grams per animal per day over a 12-day period. Data are presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001; one-way ANOVA compared with Semaglutide group followed by Dunnett's multiple comparison test.

[0038] Figure 6 : Effects of Compound B alone or in combination with Compound I on (A) relative (percent) body weight, (B) food intake (grams), (C) cumulative food intake (grams), and (D) area under the curve of Figure B in DIO mice. DIO animals were dosed once daily with different doses of Compound B alone or in combination with Compound I. Relative body weight was measured daily as % of baseline body weight of each animal over a 28-day period. Food intake was measured in grams per animal per day over an 11-day period. Data are presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001; compared with one-way ANOVA of the Compound I group, followed by Dunnett's multiple comparison test.

[0039] Figure 7: Effects of Compound E alone or in combination with semaglutide on (A) relative (percent) body weight, (B) food intake (grams), (C) cumulative food intake (grams), and (D) area under the curve of panel B in DIO mice. DIO animals were dosed once daily with different doses of Compound E alone or in combination with semaglutide. Relative body weight was measured daily as % of baseline body weight of each animal over a 28-day period. Food intake was measured in grams per animal per day over a 12-day period. Data are presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001; compared with semaglutide group by one-way ANOVA followed by Dunnett's multiple comparison test.

[0040] Figure 8 : Effects of Compound E alone or in combination with Compound I on (A) relative (percent) body weight, (B) food intake (grams), (C) cumulative food intake (grams), and (D) area under the curve of Figure B in DIO mice. DIO animals were dosed once daily with different doses of Compound E alone or in combination with Compound I. Relative body weight was measured daily as % of baseline body weight of each animal over a 28-day period. Food intake was measured in grams per animal per day over an 11-day period. Data are presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001; compared with one-way ANOVA of the Compound I group, followed by Dunnett's multiple comparison test.

[0041] Fig. 9 : Effects of Compound K alone or in combination with semaglutide on (A) relative (percent) body weight, (B) food intake (grams), (C) cumulative food intake (grams), and (D) area under the curve of panel B in DIO mice. DIO animals were dosed once daily with different doses of Compound K. Relative body weight was measured daily as % of baseline body weight of each animal over a 28-day period. Food intake was measured in grams per animal per day over a 12-day period. Data are presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001; one-way ANOVA compared with semaglutide group followed by Dunnett's multiple comparison test.

[0042] Fig.10: Effects of Compound K alone or in combination with Compound I on (A) relative (percent) body weight, (B) food intake (grams), (C) cumulative food intake (grams), and (D) area under the curve of Figure B in DIO mice. DIO animals received different doses of Compound K alone or in combination with Compound I, administered once daily. Relative body weight % was measured daily based on the baseline body weight of each animal during a 28-day period. Food intake was measured in grams for each animal daily during a 12-day period. Data are presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001; compared with one-way ANOVA of the Compound I group, followed by Dunnett's multiple comparison test. DETAILED DESCRIPTION

[0043] Additional abbreviations:

[0044] gGlu or γGlu: L-γ-glutamyl

[0045] iVal: 3-methylbutyryl (isovalerianoyl)

[0046] C18DA: 17-carboxy-heptadecanoyl (HOOC-(CH2) 16 -CO-)

[0047] The synergistic improvement in therapeutic outcome may be greater than the additive improvement in overall efficacy achieved by the combined administration compared to administration of either agent alone.

[0048] "Combination therapy" generally refers to the simultaneous or sequential administration of two or more active ingredients, so that the concentration of the individual active ingredients in the body is high enough to show a synergistic effect. The combination therapy according to the present invention can be carried out with or without instructions for combined use. Therefore, the two active ingredients can be administered completely independently or can be completely independent pharmaceutical dosage forms. The individual active ingredients may be pharmaceutical compositions that are also sold independently of each other, and only the instructions for their combined use are provided in a packaging device (such as a brochure or the like), or provided in other information (such as oral communication, written communication or the like) provided to physicians and medical staff, so as to be used simultaneously or sequentially to present a combined activity. It may refer to a fixed combination in the form of a dosage unit, or a component kit for combined administration, wherein Compound I and a compound selected from the group consisting of compounds A to L (or semaglutide and a compound selected from the group consisting of compounds A to L) can be administered independently at the same time or separately at time intervals, especially when such time intervals allow the combined object to show a cooperative (synergistic) effect.

[0049] As used herein, the term "combination administration" or "combination administration" or the like is intended to include administration of individual active ingredients to a single individual (e.g., a patient) in need thereof, and is intended to include treatment regimens in which the agents are not necessarily administered at the same time or by the same route of administration. These terms also include methods of treating a disease in which two or more active ingredients are administered sequentially so that the concentration of the individual active ingredients in the body at a given time is high enough to show a synergistic effect.

[0050] The term "fixed combination" means that the active ingredients, e.g. Compound I and a compound selected from the group consisting of Compounds A to L (or semaglutide and a compound selected from the group consisting of Compounds A to L) are both administered to a patient simultaneously in a single entity or administration form. In other terms: the active ingredients are present in one dosage form, for example in one tablet, in one prefilled syringe or in one injection device (such as an autoinjector or pen).

[0051] The present invention relates to a combination therapy comprising administering a long-acting GLP-1 / glucagon receptor dual agonist and a long-acting NPY2 receptor agonist.

[0052] More specifically, the present invention relates to a combination therapy comprising administering to a patient:

[0053] - An effective amount of Compound I:

[0054] H-His-Ac4c-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Arg-Ala-Ala-Lys-Asp-Phe-Ile-Lys(HOOC-(CH2) 16 -CO-γGlu-Gly-Ser-Gly-Ser-Gly-Gly-)-Trp-Leu-Glu-Ser-Ala-NH2 (SEQ ID NO: 01, SEQ ID NO: 02);

[0055]

[0056] and

[0057] - A long-acting NPY2 receptor agonist selected from the group consisting of compounds A to L.

[0058] Compound A:

[0059] N{α-4}-(3-methylbutyryl)-N{ε-7}-[2-(2-{2-[2-(2-{2-[(4S)-4-carboxy-4-(17-carboxyheptadecanamido)butyryl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)acetamido]-[4A,6A,7K,9E,13T,18Q,22V,28Y,30W,31L]hPYY(4-36);

[0060] iVal-APAK(C18DA-gGlu-OEG-OEG-)

[0061] PEEDATPEELQRYYVSLRHYYNWLTRQRY-NH2(SEQ ID NO:03);

[0062]

[0063] Compound B:

[0064] N{α-4}-(3-methylbutyryl)-N{ε-7}-[2-(2-{2-[2-(2-{2-[(4S)-4-carboxy-4-(17-carboxyheptadecanamido)butyryl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)acetamido]-[4A,6A,7K,9E,18Q,22V,23A,28Y,30W,31L]hPYY(4-36);

[0065] iVal-APAK(C18DA-gGlu-OEG-OEG-)

[0066] PEEDASPEELQRYYVALRHYYNWLTRQRY-NH2(SEQ ID NO:04);

[0067]

[0068] Compound C:

[0069] N{α-4}-(3-methylbutyryl)-N{ε-7}-[2-(2-{2-[2-(2-{2-[(4S)-4-carboxy-4-(17-carboxyheptadecanamido)butyryl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)acetamido]-[4A,6A,7K,9E,11E,18Q,22V,28Y,30W,31L]hPYY(4-36);

[0070] iVal-APAK(C18DA-gGlu-OEG-OEG-)

[0071] PEEEASPEELQRYYVSLRHYYNWLTRQRY-NH2(SEQ ID NO:05);

[0072]

[0073] Compound D:

[0074] N{α-4}-(3-methylbutyryl)-N{ε-7}-[2-(2-{2-[2-(2-{2-[(4S)-4-carboxy-4-(17-carboxyheptadecanamido)butyryl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)acetamido]-[4A,7K,9E,14A,18Q,22V,23A,28Y,30W,31L]hPYY(4-36);

[0075] iVal-APEK(C18DA-gGlu-OEG-OEG-)

[0076] PEEDASAEELQRYYVALRHYYNWLTRQRY-NH2 (SEQ ID NO:06);

[0077]

[0078] Compound E:

[0079] N{α-4}-(3-methylbutyryl)-N{ε-7}-[2-(2-{2-[2-(2-{2-[(4S)-4-carboxy-4-(17-carboxyheptadecanamido)butyryl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)acetamido]-[4A,6A,7K,9E,17I,18Q,22V,28Y,30W,31L]hPYY(4-36);

[0080] iVal-APAK(C18DA-gGlu-OEG-OEG-)

[0081] PEEDASPEEIQRYYVSLRHYYNWLTRQRY-NH2(SEQ ID NO:07);

[0082]

[0083] Compound F:

[0084] N{α-4}-(3-methylbutyryl)-N{ε-7}-[2-(2-{2-[2-(2-{2-[(4S)-4-carboxy-4-(17-carboxyheptadecanamido)butyryl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)acetamido]-[4A,7K,9E,10A,13T,18Q,22V,28Y,30W,31L]hPYY(4-36);

[0085] iVal-APEK(C18DA-gGlu-OEG-OEG-)

[0086] PEADATPEELQRYYVSLRHYYNWLTRQRY-NH2(SEQ ID NO:08);

[0087]

[0088] Compound G:

[0089] N{α-4}-(3-methylbutyryl)-N{ε-7}-[2-(2-{2-[2-(2-{2-[(4S)-4-carboxy-4-(17-carboxyheptadecanamido)butyryl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)acetamido]-[4A,7K,9E,13T,17I,18Q,22V,28Y,30W,31L]hPYY(4-36);

[0090] iVal-APEK(C18DA-gGlu-OEG-OEG-)

[0091] PEEDATPEEIQRYYVSLRHYYNWLTRQRY-NH2(SEQ ID NO:09);

[0092]

[0093] Compound H:

[0094] N{α-4}-(3-methylbutyryl)-N{ε-7}-[2-(2-{2-[2-(2-{2-[(4S)-4-carboxy-4-(17-carboxyheptadecanamido)butyryl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)acetamido]-[4A,7K,9E,13T,18Q,19K,22V,28Y,30W,31L]hPYY(4-36);

[0095] iVal-APEK(C18DA-gGlu-OEG-OEG-)

[0096] PEEDATPEELQKYYVSLRHYYNWLTRQRY-NH2(SEQ ID NO:10);

[0097]

[0098] Compound J:

[0099] N{α-4}-(3-methylbutyryl)-N{ε-7}-[2-(2-{2-[2-(2-{2-[(4S)-4-carboxy-4-(17-carboxyheptadecanamido)butyryl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)acetamido]-[4A,7K,9E,10A,17I,18Q,22V,28Y,30W,31L]hPYY(4-36);

[0100] iVal-APEK(C18DA-gGlu-OEG-OEG-)

[0101] PEADASPEEIQRYYVSLRHYYNWLTRQRY-NH2 (SEQ ID NO: 11);

[0102]

[0103] Compound K:

[0104] N{α-4}-(3-methylbutyryl)-N{ε-7}-(6-[(4S)-4-carboxy-4-(17-carboxyheptadecanamido)butyryl]hexanoyl)-[4A,7K,9E,10A,13T,17I,18Q,22V,28Y,30W,31L]-hPYY(4-36);

[0105] iVal-APEK(C18DA-gGlu-Ahx)PEADATPEEIQRYYVSLRHYYNWLTRQRY-NH2 (SEQ ID NO: 12);

[0106]

[0107] Compound L:

[0108] N{α-4}-(3-methylbutyryl)-N{ε-7}-(6-[(4S)-4-carboxy-4-(17-carboxyheptadecanamido)butyryl]hexanoyl)-[4A,7K,9E,13T,17I,18Q,22V,23A,28Y,30W,31L]-hPYY(4-36);

[0109] iVal-APEK(C18DA-gGlu-Ahx)PEEDATPEEIQRYYVALRHYYNWLTRQRY-NH2 (SEQ ID NO: 13);

[0110]

[0111] In an alternative embodiment, the present invention relates to a combination therapy comprising administering to a patient: - an effective amount of semaglutide:

[0112]

[0113] (N-ε26-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][Aib8,Arg34]GLP-1-(7-37), (SEQ ID NO: 17))

[0114] and

[0115] - A long-acting NPY2 receptor agonist selected from the group consisting of compounds A to L.

[0116] WO2021 / 094259 discloses the Ki measured in the radioligand binding (RLB) assay of compounds A to J (Example 1), and the effects on acute food intake (AFI) in normal NMRI mice (Example 6).

[0117] WO2022 / 029231 discloses the Ki as measured in the radioligand binding (RLB) assay of compounds K and L (Example 1), and the effects on acute food intake (AFI) in normal NMRI mice (Example 6).

[0118] The same radioligand binding (RLB) assay is described herein in Example 1. The Ki measured for Compounds A to L and References 1 to 3 are reported in Table 1 below.

[0119] Example 2 herein describes an experiment measuring the effects on acute food intake (AFI) in normal NMRI mice. The AFI % compared to the vehicle group is provided in Table 1 below.

[0120] Example 3 herein describes an experiment to measure the effects on body weight loss and food intake in diet-induced obese (DIO) mice.

[0121] All compounds showed good binding to the human NPY2 receptor (Example 1) and achieved strong inhibition of food intake after 24 h (Example 2). In the DIO study, the weight loss achieved by compounds A to L was in the range of 6.7% to 12.2% compared to the vehicle group (Example 3). However, the weight loss achieved by reference compounds Ref 1 to Ref 3 was only up to 2.7%. This shows that not all PPY2 analogs that bind to the NPY2 receptor and inhibit acute food intake are actually effective in reducing body weight.

[0122] DIO experiments (Example 3) showed that the combination of compound B, E or K and semaglutide had a synergistic effect on body weight ( Figure 5 A, 7A and 9A). The combination of Compound B, E or K together with Compound I also achieved synergistic weight loss ( Figure 6 A, 8A and 10A). When semaglutide is administered in combination with Compounds B, E or K, a synergistic effect or greater than additive benefit may be observed.

[0123] Compared with the combination of compound B and semaglutide ( Figure 5 B), the combination of Compound B and Compound I appears to provide a long-lasting reduction in food intake ( Figure 6 B). Compared with compound E and semaglutide ( Figure 7 B), a longer-lasting effect on food intake was also observed for the combination of Compound E and Compound I ( Figure 8 B). Compared with the combination of compound B or E and semaglutide ( Figure 5 A and 7A), the long-lasting reduction in food intake in combination with Compounds B and E and Compound I resulted in greater weight loss effects (% BW, Figure 6 A and 8A), and resulted in a significant reduction in total food intake ( Figure 6 D and 8D).

[0124] The combined DIO experiment showed that when a long-acting GLP-1 / glucagon receptor dual agonist is combined with a long-acting NPY2 receptor agonist according to the present invention, a synergistic effect can be achieved. In addition, compared to a combination comprising a long-acting GLP-1 receptor agonist (semaglutide) and a NPY2 receptor agonist, the combination according to the present invention can provide a longer-lasting effect on a significant reduction in food intake.

[0125] In one embodiment, the long-acting NPY2 receptor agonist is selected from the group consisting of Compounds B, E, K and L.

[0126] In another embodiment, the long-acting NPY2 receptor agonist is selected from the group consisting of Compounds B, E and K.

[0127] In another embodiment, the long-acting NPY2 receptor agonist is selected from the group consisting of Compounds E and K.

[0128] In another embodiment, the long-acting NPY2 receptor agonist is selected from the group consisting of Compounds B and E.

[0129] In another embodiment, the long-acting NPY2 receptor agonist is Compound B.

[0130] In another embodiment, the long-acting NPY2 receptor agonist is Compound E.

[0131] In another embodiment, the long-acting NPY2 receptor agonist is Compound K.

[0132] In one embodiment, the weight ratio between Compound I (or semaglutide) and the NPY2 receptor agonist is in the range of 1:10 to 10:1 (Compound I:NPY2 receptor agonist).

[0133] In one embodiment, the weight ratio between Compound I (or semaglutide) and the NPY2 receptor agonist is in the range of 1:5 to 5:1 (Compound I:NPY2 receptor agonist).

[0134] In one embodiment, the weight ratio between Compound I (or semaglutide) and the NPY2 receptor agonist is in the range of 1:3 to 3:1.

[0135] In one embodiment, the weight ratio between Compound I (or semaglutide) and the NPY2 receptor agonist is in the range of 1:2 to 2:1.

[0136] For example, the weight ratio between Compound I (or semaglutide) and Compound E is in the range of 1:3 to 3: 1. Thus, both compounds can be administered in a weight ratio of 1:2, 1:1 or 2:1.

[0137] For example, the weight ratio between Compound I (or semaglutide) and Compound K is in the range of 1:3 to 3: 1. Thus, both compounds can be administered in a weight ratio of 1:2, 1:1 or 2:1.

[0138] Although any dose combination may be used, doses of Compound I and the NPY2R agonist that provide synergistic or greater than additive benefits are generally used. For example, due to the synergistic behavior, lower doses of Compound I and the NPY2R agonist may be selected, thereby reducing the patient's medication burden while still producing a relevant effect on body weight. In addition, by providing a broad therapeutic window for such a combination, acceptable tolerability and safety may be achieved. In addition, lower doses reduce the API requirements, potentially resulting in economic benefits.

[0139] A low dose of the compound (eg, Compound I and / or NPY2R agonist) may be administered initially in gradually increasing doses (dose escalation) until a certain dose is achieved and maintained (maintenance dose).

[0140] The compounds may be administered once a week (once a week) or more frequently, preferably once a week.

[0141] In one embodiment, Compound I (or semaglutide) and a long-acting NPY2 receptor agonist are administered subcutaneously, for example, via two separate injections ("free combination") or via one injection, where both compounds are administered simultaneously, for example, via injection of a fixed dose combination of two agonists. In the case of two separate injections, one of the compounds may be administered first, followed immediately by the other, or there may be a time period between the two injections, such as 4, 3, 2 or 1 days. Alternatively, the time period between the injections is 12, 6, 4, 2 or 1 hour.

[0142] In another aspect, the invention relates to a PYY analogue selected from the group consisting of Compounds A to L for use together with Compound I in combination therapy.

[0143] Therefore, the present invention relates to a method of treating a human body, wherein the treatment comprises administering (a) PYY analogue selected from the group consisting of Compounds A to L and Compound I.

[0144] Therefore, the present invention relates to compound I for use together with a PYY analogue selected from the group consisting of compounds A to L in combination therapy.

[0145] In an alternative aspect, the invention relates to a PYY analogue selected from the group consisting of compounds A to L for use in combination therapy with semaglutide.

[0146] Therefore, the present invention relates to a method of treating a human body, wherein the treatment comprises administering (a) PYY analogue selected from the group consisting of compounds A to L and semaglutide.

[0147] Therefore, the present invention relates to semaglutide for use together with a PYY analogue selected from the group consisting of compounds A to L in combination therapy.

[0148] As described in further detail below, the combination therapy of the invention can be administered in addition to treatment with another drug therapy (e.g., an incretin-based therapy). In another embodiment, the combination therapy of the invention is administered without administering other incretin-based therapies.

[0149] In a more specific embodiment, the combination therapies of the present invention relate to the treatment of humans.

[0150] In an alternative embodiment, the present invention relates to a combination therapy comprising the administration of semaglutide and a long-acting NPY2 receptor agonist selected from the group consisting of compounds A to L, preferably compounds B, E, K or L.

[0151] Further embodiments include:

[0152] E1. A combination therapy comprising administering a long-acting GLP-1 / glucagon receptor dual agonist and a long-acting NPY2 receptor agonist.

[0153] E2. The combination therapy according to E1, comprising administering to the patient

[0154] - An effective amount of Compound I:

[0155] H-His-Ac4c-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Arg-Ala-Ala-Lys-Asp-Phe-Ile-Lys(HOOC-(CH2) 16 -CO-γGlu-Gly-Ser-Gly-Ser-Gly-Gly-)-Trp-Leu-Glu-Ser-Ala-NH2 (SEQ ID NO:01, SEQ ID NO:02) and

[0156] - a long-acting NPY2 receptor agonist selected from compounds A to L.

[0157] E3. The combination therapy according to embodiment E1 or E2, wherein the long-acting NPY2 receptor agonist is selected from compounds B, E, K and L.

[0158] E4. The combination therapy according to embodiment E1, E2 or E3, wherein the long-acting NPY2 receptor agonist is selected from the group consisting of compounds B, E and K.

[0159] E5. The combination therapy according to any one of the preceding embodiments E1 to E4, wherein the long-acting NPY2 receptor agonist is Compound B.

[0160] E6. The combination therapy according to any one of embodiments E1 to E4, wherein the long-acting NPY2 receptor agonist is Compound E.

[0161] E7. The combination therapy according to any one of embodiments E1 to E4, wherein the long-acting NPY2 receptor agonist is Compound K.

[0162] E8. The combination therapy according to any one of the preceding embodiments E1 to E7, wherein the compound may be administered once a week.

[0163] E9. A combination therapy according to any one of the preceding embodiments E1 to E8, wherein the therapy is directed to the treatment and / or prevention of obesity and various obesity-related conditions, diseases or comorbidities, such as type 2 diabetes, liver disease (such as NAFLD (non-alcoholic fatty liver disease) and NASH (non-alcoholic steatohepatitis)), kidney disease or cardiovascular disease.

[0164] E10. The combination therapy according to any one of the preceding embodiments E1 to E9, wherein the combination therapy produces a synergistic therapeutic effect, such as a synergistic effect on body weight, weight management or weight loss.

[0165] E11. The combination therapy according to embodiment E10, wherein the therapy produces a synergistic therapeutic effect compared to administration of Compound I alone.

[0166] E12. A long-acting GLP-1 / glucagon receptor dual agonist for use in a method of treating a disease, wherein the method comprises co-administering the long-acting GLP-1 / glucagon receptor dual antagonist with a long-acting NPY2 receptor agonist.

[0167] Treatment

[0168] The present invention relates to a combination of a PYY analogue and a dual GLP-1R / GCGR agonist according to the above-mentioned embodiments, which are suitable for the prevention and / or treatment of diseases and / or disorders associated with or modulated by NPY2, GLP-1 and GCG receptor activity, including but not limited to the treatment and / or prevention of obesity and various obesity-related disorders, diseases or comorbidities, such as type 2 diabetes, liver disease (such as NAFLD and NASH (non-alcoholic steatohepatitis)), kidney disease or cardiovascular disease.

[0169] The combination therapy described herein is particularly suitable for preventing weight gain or promoting weight loss. "Prevention" means inhibition or reduction when compared to the lack of treatment and does not necessarily mean a complete cessation of weight gain. The combination therapy can cause a reduction in food intake and / or increased energy expenditure and can have a beneficial effect on blood sugar control and / or on lipid metabolism (including but not limited to liver and circulating triglycerides and cholesterol content), respectively, and can reduce circulating LDL content and increase HDL / LDL ratio. Therefore, the combination therapy of the present invention can be used for direct or indirect therapy of any disease caused or characterized by excess weight, such as treatment and / or prevention of obesity, morbid obesity, obesity-related inflammation, obesity-related gallbladder disease and obesity-related sleep apnea. The combination therapy can also be used to prevent diseases or treat comorbidities associated with obesity caused or characterized by the following: insufficient blood sugar control or dyslipidemia, type 2 diabetes, metabolic syndrome, hypertension, atherosclerotic dyslipidemia, coronary heart disease, disease peripheral arterial disease, stroke or microvascular disease, heart failure and cancer. The combination may also be used to prevent disorders or treat comorbidities associated with obesity, such as liver disease (such as NAFLD and NASH), kidney disease, and to treat diseases of the central nervous system, such as cognitive dysfunction, depression, psychiatric disorders (including addictive behaviors (e.g., opioid addiction, overeating)), or neurodegenerative diseases (such as Alzheimer's disease or Parkinson's disease). The effects of the combination therapy in these diseases may be due to or associated with their effects on body weight or may be unrelated to them.

[0170] Combination therapy

[0171] Combination therapy of a PYY analogue and a dual-acting agonist for GLP-1 and GCG receptors may be administered together with another active agent to treat the disease or condition, such as an anti-obesity agent, an anti-diabetic agent, an agent for treating metabolic syndrome, an anti-dyslipidemia agent, an anti-hypertensive agent, a proton pump inhibitor or an anti-inflammatory agent. In this case, the active agents may be administered together or separately, for example as ingredients in the same pharmaceutical composition or formulation, or in the form of separate formulations. For example, the PYY analogue and the GLP-1 / GCGR agonist may be administered together, and the other active agent may be administered separately (e.g. as a background drug).

[0172] Thus, the combination according to the invention may be used in combination with an anti-obesity agent of known type. The anti-obesity agent may be amylin or an amylin analog, such as pramlintide or a calcitonin analog. Alternatively, the anti-obesity agent may be a lipase inhibitor (Orlistat TM), phentermine, melanin concentrating hormone receptor 1 antagonists, GDF-15 analogs, FGF-21 analogs, urocortin analogs, leptin analogs, GOAT inhibitors, ghrelin-receptor antagonists, neuromedin receptor 2 agonists, NPY4 receptor agonists, NPY5 receptor antagonists, melanocortin receptor 4 agonists, and analogs thereof. It will be appreciated that a PYY analogue may thus be administered in combination with a dual-acting agonist for GLP-1R / GCGR to individuals affected by a disorder or disease characterized by insufficient appetite control or otherwise excessive eating, such as bulimia and Prader-Willi syndrome. Clearly, the analogs may be used to treat a combination of disorders or diseases described herein.

[0173] Furthermore, the combination according to the invention may have some benefits if administered in combination with an antidiabetic agent of a known type, for example selected from SGLT2 inhibitors (i.e. inhibitors of sodium-glucose transport, such as empagliflozin), GPR40 agonists (FFAR1 / FFA1 agonists) or insulin or insulin analogs. Examples of suitable insulin analogs include, but are not limited to, Lantus TM 、Novorapid TM , Humalog TM 、Novomix TM 、Actraphane TM HM, Levemir TM Degludec TM and Apidra TM .

[0174] The above-mentioned combination of the present invention between PYY analogs and dual agonists for GLP-1R and GCGR can be further used in combination with drugs for the treatment of hypertension, dyslipidemia, inflammation and platelet function targeting cardiovascular diseases. Drugs for the treatment of hypertension can be selected from the group including (but not limited to) the following: angiotensin converting enzyme inhibitors, angiotensin II receptor blockers, diuretics, β-blockers or calcium channel blockers. Drugs for the treatment of heart failure can be selected from the following categories: angiotensin-receptor-neprilysin-inhibitors (ARNi), SGLT2 inhibitors (such as empagliflozin), soluble guanylate cyclase stimulators or activators (such as vericiguat), β-blockers, angiotensin converting enzyme inhibitors, angiotensin II receptor blockers or steroids (spironolactone) and non-steroids (such as finenerenone) mineralocorticoid receptor antagonists and aldosterone-synthase inhibitors.

[0175] The above-mentioned combinations of the present invention between PYY analogs and dual agonists for GLP-1R and GCGR may still further be used in combination with known types of anti-dyslipidemic agents, including but not limited to statins, fibrates, niacin, PSCK9 (proprotein convertase subtilisin type 9 / kexin) inhibitors or cholesterol absorption inhibitors.

[0176] The above-mentioned combinations of the invention between PYY analogues and dual agonists for GLP-1R and GCGR can also be combined with proton pump inhibitors of known type (i.e. with a cytotoxic agent as H + / K + -ATPase inhibitors) are used in combination with proton pump inhibitors, including (but not limited to) benzimidazole derivatives or imidazopyridine derivatives, such as Omeprazole TM .

[0177] In addition, with regard to anti-inflammatory treatment, if used with known types of anti-inflammatory agents, including but not limited to: steroids and corticosteroids (e.g., prednisone, dexamethasone), nonsteroidal anti-inflammatory agents (NSAIDs), such as propionic acid derivatives (e.g., ibuprofen); acetic acid derivatives (e.g., indomethacin, diclofenac); fenamic acid derivatives (e.g., flufenamic acid, meclofenamic acid); biphenylcarboxylic acid derivatives (e.g., diflunisal and flufenisal); oxicams (e.g., isoxicam); salicylates (e.g., acetylsalicylic acid); and pyrazolones (e.g., apazone, bezpiperylon); COX The above-mentioned combination of the present invention between a PYY analogue and a dual agonist for GLP-1R and GCGR may be beneficial if the combination is administered in combination with a PYY II inhibitor (e.g. rofecoxib); a formulation of interferon beta (e.g. interferon beta-1a or interferon beta-1b); and certain other compounds, such as 5-aminosalicylic acid and its prodrugs and pharmaceutically acceptable salts.

[0178] The above-mentioned combination of the present invention between a PYY analogue and a dual agonist for GLP-1R and GCGR can be further used in combination with drugs targeting chronic kidney disease (including diabetic nephropathy). Drugs for treating chronic and diabetic nephropathy can be selected from the group including (but not limited to) the following: angiotensin converting enzyme inhibitors, angiotensin II receptor blockers, diuretics, soluble guanylate cyclase stimulators or activators, aldosterone-synthase inhibitors and SGLT2 inhibitors (e.g., empagliflozin).

[0179] Example

[0180] Compound I is disclosed as Example 13 in WO2015 / 055801 and has the following structure: HH-Ac4c-QGTFTSDYSKYLDERAAKDFI-K([17-carboxy-heptadecanoyl]-isoGlu-GSGSGG)-WLESA-NH2 (SEQ ID NO: 01, SEQ ID NO: 02)

[0181]

[0182] Compound A is disclosed as compound 106 in WO2021 / 094259.

[0183] Compound B is disclosed as compound 117 in WO2021 / 094259.

[0184] Compound C is disclosed as compound 234 in WO2021 / 094259.

[0185] Compound D is disclosed as compound 74 in WO2021 / 094259.

[0186] Compound E is disclosed as compound 23 in WO2021 / 094259.

[0187] Compound F is disclosed as compound 14 in WO2021 / 094259.

[0188] Compound G is disclosed as compound 171 in WO2021 / 094259.

[0189] Compound H is disclosed as compound 231 in WO2021 / 094259.

[0190] Compound J is disclosed as compound 53 in WO2021 / 094259.

[0191] Compound K is disclosed as compound 7 in WO2022 / 029231.

[0192] Compound L is disclosed as compound 84 in WO2022 / 029231.

[0193] Semaglutide has the following structure:

[0194]

[0195] (N-ε26-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][Aib8,Arg34]GLP-1-(7-37), (SEQ ID NO: 17))

[0196] Example 1: Radioligand Binding Competition Assay (RLB)

[0197] The filtration RLB assay was performed in a 96-well plate with a final volume of 100 μl per well. Freeze-dried test peptides were dissolved in 100% dimethyl sulfoxide (DMSO) to a 1 mM stock solution and serially diluted in assay buffer (50 mM HEPES, 5 mM MgCl2, 1 mM CaCl2, pH 7.4) containing 0.2% ovalbumin. 10 μl / well of the test peptide solution was added to the plates to produce final concentrations ranging from 1 μM to 3 pM. Subsequently, 10 μl of human 1 μM ... 125 I-PYY (1-36) (Perkin Elmer) was added to the wells to give a final concentration of 0.02 nM. Next, 80 μL of membrane (HTS066M, ChemiSCREEN TM Human neuropeptide Y2 receptor membrane preparation, CHEMICON) was added to each well to give a final protein concentration of 0.5 μg / well. The plates were sealed and incubated for 2 hours at room temperature in a plate shaker set at 400 rpm. The cells were then plated using a 96-well FilterMate 5000 on GF / C filters pre-impregnated with 0.5% poly(ethyleneimine) (PEI). TM The culture was stopped by vacuum filtration using a collector (Perkin Elmer), followed by washing four times with 300 μl / well ice-cold wash buffer (50 mM HEPES, 500 mM NaCl, pH 7.4). The filter disk was then dried at room temperature for 60 min and the bottom of the disk was sealed with UniFilter-96 adhesive film. Finally, 50 μl / well scintillation counting mixture (Microscint 20, Packard) was added and radioactivity was counted in a Packard TopCount NXT scintillation counter. IC 50The half-maximal inhibitory concentration (IC) of the agonist was calculated by nonlinear regression analysis of the sigmoidal dose-response curve. The Ki value of the binding affinity was calculated by the Cheng-Prusoff equation (Ki = IC 50 / (1+[L] / Kd), where Kd is the previously measured receptor-specific dissociation constant (for NPY2R = 0.07 nM) and [L] is 125 I-PYY(1-36) radioligand concentration.

[0198] Ki values ​​are reported in Table 1 below.

[0199] Example 2: Effects on acute food intake in normal NMRI mice

[0200] Male NMRI mice were obtained from Charles River (Charles River, Research Models & Services Germany GmbH) or JanVier (JanVier Labs, France) at 5 weeks of age. The animals were group housed with 4 mice per cage under a 12 / 12 h dark-light cycle with lights off at 3 PM. The room temperature was controlled at 21°C ± 1°C and the humidity was 60% ± 20%. Animals had ad libitum access to regular rodent chow (KLIBA Nafag 3430 or Altromin 1324, Brogaarden, Denmark) and tap water.

[0201] Animals were transferred to a real-time food intake monitoring system, the HM-2 system (MBRose, Denmark) 5 to 7 days before the start of the study to allow adaptation to experimental conditions. Since the animals were uniquely identified with a microchip, each individual animal was identified by its microchip as soon as it entered and left the food channel. The randomization of mice in each study group (n = 7 to 8) was based on the body weight measured one day before the start of the study. The vehicle-treated (50 mM phosphate buffer pH 7 with 5% mannitol) group was included in each experiment. The animals were fasted six hours before the start of the night phase. The test peptide was administered subcutaneously to the animals once (10 nmol / kg) one hour before the night phase. Food intake was reported every hour over a 24h period. The food intake of the treatment groups was normalized (in %) by the mean food intake of the group receiving the vehicle (Table 1). Statistical significance was assessed using a one-way analysis of variance with a Turkey's multiple comparison test. P < 0.05 was considered statistically significant.

[0202] Example 3: Effects on weight loss in wild-type diet-induced obese mice

[0203] Male C57BL6 / J pre-fed with a 60% high fat diet were obtained from Jackson Laboratories at >16 weeks of age. Diet-induced obese (DIO) mice were housed individually upon arrival to obtain accurate and individual food intake measurements for each animal and assigned animal numbers. The animals were housed at room temperature of 21±2°C, relative humidity of 60±20%, and a repeated 12-h light / dark cycle (lights on at 10 am). Animals had ad libitum access to food and water throughout the study.

[0204] Before the start of treatment, stratified randomization was performed based on body weight measured one week before the start of the study. At the start of the study, the mice were approximately 20 weeks old. Body weight and food intake were measured daily before compound administration. Depending on the experiment, animals were dosed by subcutaneous injection of 30 nmol / kg once daily approximately one hour before the start of the night phase for 5 to 28 days. Control animals were dosed by daily subcutaneous injection of vehicle.

[0205] RLB analysis (NPY2R)

[0206] Table 1

[0207]

[0208]

[0209] Reference 1:

[0210] iVal-APEK(C18DA-gGlu-OEG-OEG)

[0211] PEEDASPEEIQQYYVSLRHYYNWLTRQRY-NH2 (SEQ ID NO: 14) Reference 1 is disclosed as compound 11 in WO2021 / 094259.

[0212] Reference 2:

[0213] iVal-APEK(C18DA-gGlu-OEG-OEG)

[0214] PGEDASPEELQRYYVSLRHYYHWLTRQRY-NH2(SEQ ID NO:15) Reference 3:

[0215] iVal-RPEK(C18DA-gGlu-OEG-OEG)

[0216] PEEDASPEELQRYYVSLRHYYNWLTRQRY-NH2(SEQ ID NO:16)

Claims

1. Compound I for use in a method for treating a disease, wherein the method comprises: - Compound I: H-His-Ac4c-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Arg-Ala-Ala-Lys-Asp-Phe-Ile-Lys(HOOC-(CH2) 16 -CO-γGlu-Gly-Ser-Gly-Ser-Gly-Gly-)-Trp-Leu-Glu-Ser-Ala-NH2(SEQ ID NO:01, SEQ ID NO:02) and The compound is administered in combination with a long-acting NPY2 receptor agonist selected from the group consisting of compounds A to L.

2. Compound I for use in a method for treating a disease according to claim 1, wherein the long-acting NPY2 receptor agonist is selected from the group consisting of compounds B, E, K and L.

3. Compound I for use in a method for treating a disease according to claim 1, wherein the long-acting NPY2 receptor agonist is selected from the group consisting of compounds B, E and K.

4. Compound I for use in a method for treating a disease according to claim 1, wherein the long-acting NPY2 receptor agonist is Compound B.

5. Compound I for use in a method for treating a disease according to claim 1, wherein the long-acting NPY2 receptor agonist is Compound E.

6. The compound I for use in a method for treating a disease according to claim 1, wherein the long-acting NPY2 receptor agonist is compound K.

7. Compound I for use in a method for treating a disease according to any one of claims 1 to 6, wherein the combination or the compound can be administered once a week.

8. Compound I for use in a method for treating a disease according to any one of the preceding claims, wherein the therapy is directed to the treatment and / or prevention of obesity and various obesity-related conditions, diseases or comorbidities, such as type 2 diabetes, liver diseases such as NAFLD (non-alcoholic fatty liver disease) and NASH (non-alcoholic steatohepatitis), kidney disease or cardiovascular disease.

9. Compound I for use in a method for treating a disease according to any one of the preceding claims, wherein the combined administration has a synergistic effect on the patient, such as a synergistic effect on body weight, weight management or weight loss.

10. Compound I for use in a method for treating a disease according to any one of the preceding claims, wherein the use is for use in the treatment of obesity and / or obesity-related disorders, for use in the treatment of type 2 diabetes, liver disease (such as NAFLD (non-alcoholic fatty liver disease) or NASH (non-alcoholic steatohepatitis)), kidney disease or cardiovascular disease.

11. Compound I for use in a method for treating a disease according to any one of claims 1 to 9, wherein the use is for treating obesity or losing weight.

12. A pharmaceutical composition comprising - Compound I: H-His-Ac4c-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Arg-Ala-Ala-Lys-Asp-Phe-Ile-Lys(HOOC-(CH2) 16 -CO-γGlu-Gly-Ser-Gly-Ser-Gly-Gly-)-Trp-Leu-Glu-Ser-Ala-NH2(SEQ ID NO:01, SEQ ID NO:02) and - a long-acting NPY2 receptor agonist selected from compounds A to L.

13. The pharmaceutical composition according to claim 12, wherein the long-acting NPY2 receptor agonist is selected from compounds B, E and K.

14. The pharmaceutical composition according to claim 12 or 13, for use in the treatment of obesity and / or obesity-related disorders, for use in the treatment of type 2 diabetes, NAFLD (non-alcoholic fatty liver disease), NASH (non-alcoholic steatohepatitis), renal disease or cardiovascular disease.

15. The composition according to any one of claims 12 to 14, wherein the combination produces a synergistic effect, such as a synergistic effect on body weight, weight management or weight loss.

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