Polypeptide dosage regimens

Abstract

The present invention relates to a method of reducing the body weight of a subject, the method comprising administering to the subject about 1 mg to about 15 mg of an amylin receptor (AMYR) agonist. Also provided are corresponding compositions and kits.

Description

[0001] POLYPEPTIDE DOSAGE REGIMENS

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to dosage regimens for reducing body weight in a subject by administering an amylin receptor agonist.

[0004] BACKGROUND

[0005] Obesity is the result of complex relationships between genetic, socioeconomic, and cultural influences, and is defined as a BMI of > 30 kg / m2. It is a risk factor for the development of many comorbid conditions, including type 2 diabetes mellitus, CV disease, and chronic kidney disease, and it is estimated that obesity affects nearly a third of the world’s population (Chooi et al. 2019 Metabolism. 2019;92:6-10). Weight loss has been associated with improvements in comorbidities, however, achieving and maintaining weight loss through lifestyle interventions alone is difficult. Bariatric surgery is currently the most effective treatment for sustained body weight loss (O’Brien et al. 2019 Obes Surg. 2019;29(1 ):3-1 ); however, such surgical intervention is only appropriate for a minority of the patient population.

[0006] Despite the development of several new treatments for overweight and obesity, concerns regarding tolerability, suboptimal response and the need for additional weight loss persist among a significant number of patients, highlighting an unmet need for additional treatment modalities.

[0007] Amylin receptor agonists are being considered for the treatment of obesity, excess food intake, and diabetes (see e.g., WO 2018 / 046719). Pramlintide, a synthetic analogue of human amylin, is clinically used in amylin replacement therapies and simulates the important glucoregulatory actions of amylin. These glucoregulatory actions complement those of insulin by regulating the rate of appearance of glucose in the circulation, and are achieved through three primary mechanisms: slowing the rate of gastric emptying, suppression of post-meal glucagon secretion and suppression of food intake (Roth JD et. al. GLP-1 R and amylin agonism in metabolic disease: complementary mechanisms and future opportunities. Br J Pharmacol. 2012 ; 166(1 ):121 -136). Pramlintide has been used as an adjunct to insulin in patients with diabetes who have failed to reach desired glucose control despite optimal insulin therapy (Pullman J, et. al. Pramlintide is used in the management of insulin-using patients with type 2 and type 1 diabetes. Vase Health Risk Manag. 2006;2(3):203-212). Pramlintide analogues conjugated to lipids to extend their half-life are also known.

[0008] Existing agents are not without their problems. Some amylin analogues are reported to be associated with poor tolerability, as nausea and vomiting are common side effects, as well as the need for frequent administration due to a short half life.

[0009] Consequently, there remains an unmet need for medications that can achieve greater levels of weight loss and the concomitant improvements in health outcomes. It is an object of the present invention to address one or more of these issues.

[0010] SUMMARY OF THE INVENTION

[0011] AZD6234 is a potent agonist of human, cynomolgus monkey, and rat amylin receptors (AMYR), displaying in vitro selectivity over calcitonin receptor activation. In vitro studies and in vivo toxicology studies demonstrated acceptable safety profiles in rats and cynomolgus monkey. The present inventors sought to determine, for the first time, how AZD6234 behaves when administered to humans at various doses.

[0012] The inventors have established through a Phase I human clinical trial that AZD6234 is generally safe and well tolerated when administered to humans. The inventors have combined in vivo pharmacokinetic and safety data obtained from the in-human trial with preclinical efficacy data obtained from animal models to predict dosages of AZD6234 that are expected to achieve an efficacious weight reducing effect in humans without compromising patient safety. Further, it was established that AMYR agonists which have selectivity for AMYR as compared with calcitonin receptor (CTR) result in improved tolerability (e.g., reduction in nausea and / or vomiting). As such, AMYR agonists, particularly those which have selectivity for AMYR, such as AZD6234, at doses as described herein have the potential to achieve clinically beneficial weight loss without adverse clinical events and with improved tolerability compared with other anti-obesity drugs.

[0013] Hence, administering an AMYR agonist, such as AZD6234, at the doses described herein is expected to achieve clinical benefit when administered to humans for reducing body weight, including the treatment of obesity and obesity-related conditions. Such treatments may be particularly suitable for treating patients who are unsuitable for, or unable to tolerate, treatment with other weight loss actives, such as GLP-1 receptor (GLP-1 R) agonists (e.g. semaglutide) and / or GLP-1 R / glucagon receptor (GCGR) dual-agonists (e.g. tirzepatide), particularly monotherapies using such actives.

[0014] Accordingly, the present invention provides a method of reducing the body weight of a subject, the method comprising administering to the subject about 1 mg to about 15 mg of an amylin receptor (AMYR) agonist.

[0015] The amount of AMYR agonist may be about 1 mg to about 10 mg.

[0016] The amount of AMYR agonist may be about 1 mg to about 5 mg.

[0017] The amount of AMYR agonist may be about 4 mg to about 10 mg.

[0018] The amount of AMYR agonist may be about 1 mg to about 3 mg.

[0019] The amount of AMYR agonist may be about 2.5 mg to about 5 mg.

[0020] The amount of AMYR agonist may be about 7 mg to about 10 mg.

[0021] The amount of AMYR agonist may be about 1.5 mg, about 1.6 mg, about 4.5 mg, about 6 mg, or about 9 mg.

[0022] The amount of AMYR agonist may be about 1 .5 mg, about 4.5 mg, or about 9 mg.

[0023] In any method of the invention, the AMYR agonist may be administered by subcutaneous injection.

[0024] In any method of the invention, the AMYR agonist may be administered by selfadministration.

[0025] In any method of the invention, the AMYR agonist may be administered about once every 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, or 14 days; optionally the AMYR agonist may be administered about once a week.

[0026] In any method of the invention, the AMYR agonist may be administered to the subject multiple times over a period of at least 3 months, 6 months, 9 months, 1 year, 2 years, or 5 years.

[0027] In any method of the invention, the AMYR agonist may be titrated to a final dose over a period between about 4 weeks and about 8 weeks. In any method of the invention, the subject’s body weight may be reduced by >5%, optionally, the subject’s body weight may be reduced by >10%, >15%, or >20%.

[0028] In any method of the invention, the subject’s reduction in body weight may be fat-specific weight loss.

[0029] In any method of the invention, the subject may experience reduced nausea as compared to a therapeutically effective dose of a dual amylin-calcitonin receptor agonist (DACRA) or a GLP-1 R agonist.

[0030] The AMYR agonist may have selectivity to AMYR as compared to a calcitonin receptor (CTR).

[0031] The AMYR agonist may have at least a 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 12-fold, at least 15-fold, at least 17-fold, at least 20-fold, or at least 25-fold selectivity to AMYR as compared to CTR; optionally the AMYR agonist may have at least a 10-fold selectivity to AMYR as compared to CTR.

[0032] The AMYR agonist may be selected from a polypeptide, small molecule drug, antibody, antibody-drug conjugate, or aptamer; or a pharmaceutically acceptable salt thereof.

[0033] The AMYR agonist may be a polypeptide, or a pharmaceutically acceptable salt thereof.

[0034] The AMYR may be a human AMYR. The AMYR may be AMY1 R, AMY2R and / or AMY3R.

[0035] The AMYR agonist may be an AMYR agonist polypeptide, or a pharmaceutically acceptable salt thereof, which comprises an amino acid sequence having at least 90% identity to pramlintide (KCNTATCATQRLANFLVHSSNNFGPILPPTNVGSNTY-amide, SEQ ID NO: 5).

[0036] The AMYR agonist polypeptide may be lipidated and / or the lipid may be attached to an amino acid residue in the AMYR agonist polypeptide by a linker.

[0037] The AMYR agonist polypeptide may comprise or consist of an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 99%, or 100% identity to the amino acid sequence K[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 26).

[0038] The AMYR agonist polypeptide, or pharmaceutically acceptable salt thereof, may comprise or consist of the amino acid sequence K(yE-yE- C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 11).

[0039] In any method of the invention, the subject may have a disease or disorder selected from obesity, metabolic disease, an obesity-related condition, eating disorder, Alzheimer's disease, hepatic steatosis ("fatty liver"), kidney failure, arteriosclerosis (e.g. atherosclerosis), cardiovascular disease, macrovascular disease, microvascular disease, diabetic heart (including diabetic cardiomyopathy and heart failure as a diabetic complication), coronary heart disease, peripheral artery disease or stroke, cancer, dumping syndrome, hypertension e.g. pulmonary hypertension, or dyslipidemia e.g. atherogenic dyslipidemia, cholescystitis, or short bowel syndrome. The subject may have obesity.

[0040] The subject may have an obesity-related condition. The obesity-related condition may be overweight, morbid obesity, obesity prior to surgery, obesity-linked inflammation, obesity- linked gallbladder disease, sleep apnoea and respiratory problems, hyperlipidaemia, degeneration of cartilage, osteoarthritis, or reproductive health complications of obesity or overweight such as infertility. The subject may be overweight. The subject may be overweight with at least one comorbidity selected from hypertension, dyslipidaemia, cardiovascular (CV) disease, or obstructive sleep apnoea.

[0041] The subject may have a metabolic disease, optionally wherein the metabolic disease includes diabetes, type 1 diabetes, type 2 diabetes, gestational diabetes, pre-diabetes, insulin resistance, impaired glucose tolerance (IGI), disease states associated with elevated blood glucose levels, metabolic syndrome, or hyperglycaemia (e.g. abnormal postprandial hyperglycaemia).

[0042] The AMYR agonist may be a polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence K(yE-yE-

[0043] C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 11 ) in an amount of about 1 .5 mg, about 4.5 mg, or about 9 mg, and wherein the subject has obesity or an obesity-related condition.

[0044] The invention also provides a method of treating and / or preventing obesity or an obesity- related condition in a subject in need thereof, the method comprising administering to the subject about 1 mg to about 15 mg of an amylin receptor (AMYR) agonist.

[0045] The invention also provides a method of treating and / or preventing a metabolic disease in a subject in need thereof, the method comprising administering to the subject about 1 mg to about 15 mg of an amylin receptor (AMYR) agonist. The invention also provides a cosmetic method of reducing the body weight of a subject, the method comprising administering to the subject about 1 mg to about 15 mg of an amylin receptor (AMYR) agonist.

[0046] The invention also provides an AMYR agonist for use in a method of treating and / or preventing obesity or an obesity-related condition in a subject in need thereof, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist.

[0047] The invention also provides an AMYR agonist for use in a method of treating and / or preventing a metabolic disease in a subject in need thereof, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist.

[0048] The invention also provides an AMYR agonist for use in a method of reducing body weight in a subject, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist.

[0049] In any method of the invention, the subject has a disease or disorder selected from obesity, an obesity-related condition, and metabolic disease.

[0050] The invention also provides a kit comprising (a) an AMYR agonist, and (b) instructions for use of the same for treating or preventing obesity or an obesity-related condition at a dose of about 1 mg to about 15 mg.

[0051] BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1 shows that an AMYR agonist demonstrated dose dependent weight loss following a single dose of either 0.3 mg, 0.9 mg, 1.5 mg, 2.7 mg, or 4.2 mg AMYR agonist in a Phase I trial of healthy subjects who are overweight or obese.

[0053] Figure 2 shows that administration of the AMYR agonist demonstrated a good tolerability profile in the Phase I trial, in which no safety concerns were identified at single doses up to 4.2 mg.

[0054] Figure 3 shows that sustained weight loss was achieved following (A) six weeks of repeat dosing (once weekly) of 2.7 mg AMYR agonist (s.c.) and (B) twelve weeks of repeat dosing (once weekly) gradually titrated from 1.5 mg to 9 mg AMYR agonist (s.c.) in preliminary data from a Phase lb trial of healthy subjects who are overweight or obese. Figure 4 shows that good tolerability was achieved (A) on repeat dosing of 2.7 mg AMYR agonist (s.c.) and (B) on repeat dosing of AMYR agonist (s.c.) gradually titrated from 1 .5 mg to 9 mg in preliminary data from the Phase lb trial.

[0055] Figure 5 shows a simplified layout of a Phase II double-blind, placebo-controlled study of an AMYR agonist at doses of 1.5 mg, 4.5 mg and 9 mg in participants living with obesity or overweight with comorbidity.

[0056] Figure 6 shows: (A) concentration of plasma endogenous leptin following a 4h fast in DIO and lean rats treated with AZD6234 or Semaglutide; (B) immunohistochemistry of pSTAT in the VHM of rats from (A); (C) bar graph quantifying the %pSTAT3 signalling in the mice of (A) challenged with PBS or a murine leptin challenge (13 mg / kg). In the right-hand panel, the left bar for each group represents PBS challenge and the right bar for each group represents leptin challenge. # = Student paired T test * = One way ANOVA

[0057] Figure 7 shows: Acute effects of single AMYR agonist therapy on acute food intake in lean rats. (A) 72-hour cumulative food intake. (B) Food intake during dark cycles. One-way ANOVA, Dunnett analysis. *p<0.05, ** p<0.01 vs vehicle.

[0058] Figure 8 shows: Effects of repeat AMYR agonist therapy on body weight and body composition in DIO rats. (A) Body weight change normalized to vehicle. Two-way ANOVA, Dunnett’s multiple comparison test. (B) Body composition change in fat mass and lean mass (g) compared to vehicle. One-way ANOVA, Turkey’s multiple comparison test. *p<0.05, ** p<0.01 , ***p<0.001 , ****p<0.0001 . (C) Lean / fat mass ratio time course over the study.

[0059] Figure 9: DIO rats were dosed QD for 4 weeks with either Vehicle, AMYR agonist (2.5 or 10 nmol / kg), or DACRA (1 , 3, 10 or 20 nmol / kg). (A) Graph showing body weight change (%) normalized to vehicle over time. (B). Body composition change (fat mass (L) and fat-free mass (R), g) at end of study. (C) Total body weight change (%) normalized to vehicle and corresponding proportion of fat mass loss and fat-free mass-loss at end of study. (D) Emax model showing relationship between calcitonin receptor engagement with lean-mass change (g, corrected for control) (L) and amylin receptor engagement with fat-mass change (g, corrected for control) (R).

[0060] Figure 10: (A) Graph showing result of saccharin conditioned tasted aversion dose response experiment for AZD6234 and a DACRA. Data reported as mean ± standard error of the mean (SEM); data analysis performed using Graph Pad Prism - statistical analysis performed using One-way ANOVA with Tukey ad hoc analysis. (*p<0.05, **p<0.01 , ***0<0.001 , ****p<0.0001 vs. Vehicle). (B) Emax model showing relationship between calcitonin receptor engagement (L) or amylin receptor engagement (R) (fold in vitro potency normalised for fraction unbound) with saccharine aversion in lean rats.

[0061] Figure 11 : Lean Han Wistar rats were dosed with single s.c. administration of AMYR agonist or vehicle and subsequently administered barium sulphate HPMC contrast to evaluate gastric emptying in a 2-hour study. Graph showing time course of gastric emptying (%) following administration of tested compounds.

[0062] DETAILED DESCRIPTION

[0063] Definitions

[0064] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Singleton, et al., DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY, 20 ED., John Wiley and Sons, New York (1994), and Hale & Marham, THE HARPER COLLINS DICTIONARY OF BIOLOGY, Harper Perennial, NY (1991) provide the skilled person with a general dictionary of many of the terms used in this disclosure.

[0065] This disclosure is not limited by the exemplary methods and materials disclosed herein, and any methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of this disclosure. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention, which is defined solely by the claims.

[0066] The description of embodiments of the disclosure is not intended to be exhaustive or to limit the disclosure to the precise form disclosed. While specific embodiments of, and examples for, the disclosure are described herein for illustrative purposes, various equivalent modifications are possible within the scope of the disclosure, as those skilled in the relevant art will recognise. For example, while method steps or functions are presented in a given order, alternative embodiments may perform functions in a different order, or functions may be performed substantially concurrently. The teachings of the disclosure provided herein can be applied to other procedures or methods as appropriate. The various embodiments described herein can be combined to provide further embodiments. Aspects of the disclosure can be modified, if necessary, to employ the compositions, functions and concepts of the above references and application to provide yet further embodiments of the disclosure. Moreover, due to biological functional equivalency considerations, some changes can be made in protein structure without affecting the biological or chemical action in kind or amount. These and other changes can be made to the disclosure in light of the detailed description. All such modifications are intended to be included within the scope of the appended claims.

[0067] The headings provided herein are not limitations of the various aspects or embodiments of this disclosure.

[0068] As used herein, the term "capable of' when used with a verb, encompasses, or means the action of the corresponding verb. For example, "capable of activating" also means activates, "capable of agonising" also means agonises, "capable of binding" also means binds and "capable of specifically activating..." also means specifically activates.

[0069] Numeric ranges are inclusive of the numbers defining the range. Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limits of that range is also specifically disclosed. Each smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in that stated range is encompassed within this disclosure. The upper and lower limits of these smaller ranges may independently be included or excluded in the range, and each range where either, neither or both limits are included in the smaller ranges is also encompassed within this disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in this disclosure.

[0070] As used herein, the articles "a" and “an” may refer to one or to more than one (e.g., to at least one) of the grammatical object of the article. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. In this application, the use of "or" means "and / or" unless stated otherwise. Furthermore, the use of the term "including", as well as other forms, such as "includes" and "included", is not limiting.

[0071] “About” may generally mean an acceptable degree of error for the quantity measured given the nature or precision of the measurements. Exemplary degrees of error are within 20 percent (%), typically, within 10%, and more typically, within 5% of a given value or range of values. Preferably, the term “about” shall be understood herein as plus or minus (±) 5%, preferably ± 4%, ± 3%, ± 2%, ± 1%, ± 0.5%, ± 0.1%, of the numerical value of the number with which it is being used.

[0072] The term "consisting of" refers to compositions, methods, and respective components thereof as described herein, which are exclusive of any element not recited in that description of the invention.

[0073] As used herein the term "consisting essentially of" refers to those elements required for a given invention. The term permits the presence of elements that do not materially affect the basic and novel or functional characteristic(s) of that invention (i.e., inactive, or non- immunogenic ingredients).

[0074] Embodiments described herein as “comprising” one or more features may also be considered as disclosure of the corresponding embodiments “consisting of” and / or “consisting essentially of” such features, of the corresponding embodiments "consisting of" such features.

[0075] Amino acids are referred to herein using the name of the amino acid, the three-letter abbreviation, or the single letter abbreviation. Unless otherwise indicated, amino acid sequences are written left to right in amino to carboxy orientation.

[0076] The term “protein", as used herein, includes proteins, polypeptides, and peptides. As used herein, the term “amino acid sequence” is synonymous with the term “polypeptide” and / or the term “protein”. In some instances, the term “amino acid sequence” is synonymous with the term “peptide”. The terms "protein" and "polypeptide" are used interchangeably herein. In the present disclosure and claims, the conventional one-letter and three-letter codes for amino acid residues may be used. The 3-letter code for amino acids as defined in conformity with the IUPACIUB Joint Commission on Biochemical Nomenclature (JCBN). It is also understood that a polypeptide may be coded for by more than one nucleotide sequence due to the degeneracy of the genetic code.

[0077] A “fragment” of a polypeptide typically comprises at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 97% or more of the original polypeptide. A “variant” amino acid sequence has substantial homology or substantial similarity to a reference amino acid sequence (or a fragment thereof). An amino acid sequence or fragment thereof is “substantially homologous” (or “substantially identical”) to a reference sequence if, when optimally aligned (with appropriate amino acid insertions or deletions) with the other amino acid (or its complementary strand), there is nucleotide sequence identity in at least about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more % of the amino acids. Methods for homology determination of amino acid sequences are known in the art. Typically, a variant polypeptide of the invention retains the function or activity of the full-length polypeptide.

[0078] A variant polypeptide may be one in which amino acid residues from one species are substituted for the corresponding residue in another species, either at the conserved or nonconserved positions. Variants of amylin receptor agonists and / or GLP-1 R agonists disclosed herein may be produced and used in the present invention. Following the lead of computational chemistry in applying multivariate data analysis techniques to the structure / property-activity relationships [see for example, Wold, et al. Multivariate data analysis in chemistry. Chemometrics-Mathematics and Statistics in Chemistry (Ed.: B. Kowalski); D. Reidel Publishing Company, Dordrecht, Holland, 1984 (ISBN 90-277-1846-6] quantitative activity-property relationships of DNMTs can be derived using well-known mathematical techniques, such as statistical regression, pattern recognition and classification [see for example Norman et al. Applied Regression Analysis. Wiley- Interscience; 3rd edition (April 1998) ISBN: 0471170828; Kandel, Abraham et al. Computer- Assisted Reasoning in Cluster Analysis. Prentice Hall PTR, (May 11 , 1995), ISBN: 0133418847; Krzanowski, Wojtek. Principles of Multivariate Analysis: A User's Perspective (Oxford Statistical Science Series, No 22 (Paper)). Oxford University Press; (December 2000), ISBN: 0198507089; Witten, Ian H. et al Data Mining: Practical Machine Learning Tools and Techniques with Java Implementations. Morgan Kaufmann; (October 11 , 1999), ISBN:1558605525; Denison David G. T. (Editor) et al Bayesian Methods for Nonlinear Classification and Regression (Wiley Series in Probability and Statistics). John Wiley & Sons; (July 2002), ISBN: 0471490369; Ghose, Arup K. et al. Combinatorial Library Design and Evaluation Principles, Software, Tools, and Applications in Drug Discovery. ISBN: 0- 8247-0487-8]. The properties of a DNMT can be derived from empirical and theoretical models (for example, analysis of likely contact residues or calculated physicochemical property) of the DNMT sequence, functional and three-dimensional structures and these properties can be considered individually and in combination. Methods of determining amino acid percentage sequence identity are known in the art. By way of example, when assessing amino acid sequence identity, a sequence having a defined number of contiguous amino acids may be aligned with a amino acid sequence (having the same number of contiguous amino acids) from the corresponding portion of a amino acid sequence of the present invention. Tools known in the art for determining amino acid percentage sequence identity include Protein BLAST.

[0079] Amino acid residues at non-conserved positions may be substituted with conservative or non-conservative residues. In particular, conservative amino acid replacements are contemplated.

[0080] A “conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art, including basic side chains (e.g., lysine, arginine, or histidine), acidic side chains (e.g., aspartic acid or glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, or cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, or tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, or histidine). Thus, if an amino acid in a polypeptide is replaced with another amino acid from the same side chain family, the amino acid substitution is considered to be conservative. The inclusion of conservatively modified variants in a DNMT of the invention does not exclude other forms of variant, for example polymorphic variants, interspecies homologs, and alleles.

[0081] “Non-conservative amino acid substitutions” include those in which (i) a residue having an electropositive side chain (e.g., Arg, His or Lys) is substituted for, or by, an electronegative residue (e.g., Glu or Asp), (ii) a hydrophilic residue (e.g., Ser or Thr) is substituted for, or by, a hydrophobic residue (e.g., Ala, Leu, lie, Phe or Vai), (iii) a cysteine or proline is substituted for, or by, any other residue, or (iv) a residue having a bulky hydrophobic or aromatic side chain (e.g., Vai, His, lie or Trp) is substituted for, or by, one having a smaller side chain (e.g., Ala or Ser) or no side chain (e.g., Gly).

[0082] As used herein, the terms “polynucleotides”, "nucleic acid" and "nucleic acid sequence" refers to any molecule, preferably a polymeric molecule, incorporating units of ribonucleic acid, deoxyribonucleic acid, or an analogue thereof. The nucleic acid can be either singlestranded or double-stranded. A single-stranded nucleic acid can be one nucleic acid strand of a denatured double- stranded DNA Alternatively, it can be a single-stranded nucleic acid not derived from any double-stranded DNA. In one aspect, the nucleic acid can be DNA. In another aspect, the nucleic acid can be RNA Suitable nucleic acid molecules are DNA, including genomic DNA or cDNA. Other examples of nucleic acid molecules are RNA, including siRNA, shRNA, and antisense oligonucleotides. Typically, the methods of the invention relate to the production of oligonucleotides (short DNA or RNA sequences typically less than about 300 bases in length).

[0083] Unless otherwise indicated, any nucleic acid sequences are written left to right in 5' to 3' orientation.

[0084] The polynucleotides of the present invention may be prepared by any means known in the art. For example, large amounts of the polynucleotides may be produced by replication in a suitable host cell. The natural or synthetic DNA fragments coding for a desired fragment will be incorporated into recombinant nucleic acid constructs, typically DNA constructs, capable of introduction into and replication in a prokaryotic or eukaryotic cell. Usually, the DNA constructs will be suitable for autonomous replication in a unicellular host, such as yeast or bacteria, but may also be intended for introduction to and integration within the genome of a cultured insect, mammalian, plant, or other eukaryotic cell lines.

[0085] The polynucleotides of the present invention may also be produced by chemical synthesis, e.g., by the phosphoramidite method or the tri-ester method and may be performed on commercial automated oligonucleotide synthesisers. A double-stranded fragment may be obtained from the single stranded product of chemical synthesis either by synthesising the complementary strand and annealing the strand together under appropriate conditions or by adding the complementary strand using DNA polymerase with an appropriate primer sequence.

[0086] In view of the degeneracy of the genetic code, considerable sequence variation is possible among the polynucleotides of the present invention. One of ordinary skill in the art will appreciate that flexibility exists when determining a degenerate codon, representative of all possible codons encoding each amino acid. For example, some polynucleotides encompassed by the degenerate sequence may encode variant amino acid sequences, but one of ordinary skill in the art can easily identify such variant sequences by reference to the amino acid sequences of the present invention.

[0087] A “variant” nucleic acid sequence has substantial homology or substantial similarity to a reference nucleic acid sequence (or a fragment thereof). A nucleic acid sequence or fragment thereof is “substantially homologous” (or “substantially identical”) to a reference sequence if, when optimally aligned (with appropriate nucleotide insertions or deletions) with the other nucleic acid (or its complementary strand), there is nucleotide sequence identity in at least about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more % of the nucleotide bases. Methods for homology determination of nucleic acid sequences are known in the art.

[0088] Alternatively, a “variant” nucleic acid sequence is substantially homologous with (or substantially identical to) a reference sequence (or a fragment thereof) if the “variant” and the reference sequence they are capable of hybridising under stringent (e.g., highly stringent) hybridisation conditions. Nucleic acid sequence hybridisation will be affected by such conditions as salt concentration (e.g. NaCI), temperature, or organic solvents, in addition to the base composition, length of the complementary strands, and the number of nucleotide base mismatches between the hybridising nucleic acids, as will be readily appreciated by those skilled in the art. Stringent temperature conditions are preferably employed, and generally include temperatures in excess of 30°C, typically in excess of 37°C and preferably in excess of 45°C. Stringent salt conditions will ordinarily be less than 1000 mM, typically less than 500 mM, and preferably less than 200 mM. The pH is typically between 7.0 and 8.3. The combination of parameters is much more important than any single parameter.

[0089] Methods of determining nucleic acid percentage sequence identity are known in the art. By way of example, when assessing nucleic acid sequence identity, a sequence having a defined number of contiguous nucleotides may be aligned with a nucleic acid sequence (having the same number of contiguous nucleotides) from the corresponding portion of a nucleic acid sequence of the present invention. Tools known in the art for determining nucleic acid percentage sequence identity include Nucleotide BLAST.

[0090] One of ordinary skill in the art appreciates that different species exhibit “preferential codon usage”. As used herein, the term “preferential codon usage” refers to codons that are most frequently used in cells of a certain species, thus favouring one or a few representatives of the possible codons encoding each amino acid. For example, the amino acid threonine (Thr) may be encoded by ACA, ACC, ACG, or ACT, but in mammalian host cells ACC is the most commonly used codon; in other species, different codons may be preferential. Preferential codons for a particular host cell species can be introduced into the polynucleotides of the present invention by a variety of methods known in the art. Introduction of preferential codon sequences into recombinant DNA can, for example, enhance production of the protein by making protein translation more efficient within a particular cell type or species. A “fragment” of a polynucleotide of interest comprises a series of consecutive nucleotides from the sequence of said full-length polynucleotide. By way of example, a “fragment” of a polynucleotide of interest may comprise (or consist of) at least 30 consecutive nucleotides from the sequence of said polynucleotide (e.g., at least 35, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800 850, 900, 950 or 1000 consecutive nucleic acid residues of said polynucleotide). Typically, a fragment as defined herein retains the same function as the full-length polynucleotide.

[0091] When applied to a nucleic acid sequence, the term “isolated” denotes that the polynucleotide sequence has been removed from its natural genetic milieu and is thus free of other extraneous or unwanted coding sequences (but may include naturally occurring 5' and 3' untranslated regions such as promoters and terminators) and is in a form suitable for use within genetically engineered protein production systems. When applied to a protein, such as an amylin receptor agonist amino acid sequence, the term “isolated” denotes that the protein has been removed from its natural cellular milieu and is thus free of other extraneous or unwanted coding proteins and / or genetic material and is in a form suitable for use within genetically engineered protein production systems. Such isolated molecules are those that are separated from their natural environment. In the context of the invention, an isolated protein nucleic acid is one which has been separated from one or more of the reagents used in its production according to methods of the invention; one which has been separated from the other proteins and / or nucleic acid sequences synthesised in the same iteration of the method and / or one which has been separated from the one or more units on which it was synthesised.

[0092] The terms "decrease", "reduce", "reduction", or "inhibit" are all used herein to mean a decrease by a statistically significant amount. The terms "reduce," "reduction" or "decrease" or "inhibit" typically means a decrease by at least 5% as compared to a reference level (e.g. the absence of a given treatment) and can include, for example, a decrease by at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or more. As used herein, "reduction" or "inhibition" encompasses a complete inhibition or reduction as compared to a reference level. "Complete inhibition" is a 100% inhibition (i.e., abrogation) as compared to a reference level. The terms "increased", "increase", "enhance", or "activate" are all used herein to mean an increase by a statically significant amount. The terms "increased", "increase", "enhance", or "activate" can mean an increase of at least 25%, at least 50% as compared to a reference level, for example an increase of at least about 50%, or at least about 75%, or at least about 80%, or at least about 90%, at least about 95%, or at least about 98%, or at least about 99%, or at least about 100%, or at least about 250% or more compared with a reference level, or at least about a 1 .5-fold, or at least about a 2-fold, or at least about a 2.5-fold, or at least about a 3-fold, or at least about a 4-fold, or at least about a 5-fold or at least about a 10-fold increase, or any increase between 1.5-fold and 10-fold or greater as compared to a reference level.

[0093] As used herein, the term “treating” or “treatment” refers to an amelioration of a disease or disorder, or at least one discernible symptom thereof. "Treatment" or "treating" may refer to an amelioration of at least one measurable physical parameter, not necessarily discernible by the patient. "Treatment" or "treating" may refer to inhibiting, reducing, or delaying the progression of a disease or disorder, either physically, e.g., stabilization of a discernible symptom, physiologically, e.g., stabilization of a physical parameter, or both. By way of nonlimiting example, “treating” or “treatment” may refer to reducing the progression of an obesity-related condition in a subject compared to the progression that would be expected for a subject having that obesity-related condition or compared to the progression that would be expected in a subject at that stage of the obesity-related condition. "Treatment" or "treating" may refer to delaying the onset of a disease or disorder.

[0094] As used herein, the term “preventing” or “prevention” refers to a reduction of the risk of acquiring a given disease or disorder. “Preventing” or “prevention” may refer to preventing or delaying the onset of one or more symptoms of a disease or disorder, e.g., compared to an untreated control or placebo.

[0095] The term “subject” refers to an animal, such as a mammal, that has been or will be the object of treatment, observation, or experiment. The methods described herein may be useful for both human therapy and veterinary applications. Preferably, the subject is a human. The subject to be treated may be a human adult. The subject may be a human and may be an adult, juvenile or infant, typically an adult.

[0096] As used herein, the term “sample” refers to a sample of biological materials (cells, tissue, fluid, etc.) obtained from an individual. The sample may be any suitable biological material, for example blood, plasma, saliva, serum, sputum, urine, cerebral spinal fluid, cells, a cellular extract, a tissue sample, a tissue biopsy, a stool sample and the like. Typically, the sample is blood sample. The precise biological sample that is taken from the individual may vary, but the sampling preferably is minimally invasive and is easily performed by conventional techniques. The sample may be a whole blood sample, a purified peripheral blood leukocyte sample or a cell type sorted leukocyte sample, such as a sample of the individual’s neutrophils.

[0097] The term “pharmaceutically acceptable” as used herein means approved by a regulatory agency of the Federal or a state government, or listed in the U.S. Pharmacopeia, European Pharmacopeia, or other generally recognised pharmacopeia.

[0098] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that such publications constitute prior art to the claims appended hereto. All documents cited herein are each entirely incorporated by reference herein, including all data, tables, figures, and text presented in the cited documents.

[0099] Treatment and Dosage Regimens

[0100] The invention relates to therapies comprising an amylin receptor (AMYR) agonist. An AMYR agonist is a molecule that is capable of binding to, and inducing signalling by, one or more receptors or receptor complexes regarded as physiological receptors for human amylin.

[0101] Treatment methods

[0102] The present invention encompasses therapies which involve administering between about 1 mg to about 15 mg of an AMYR agonist of the invention to a subject for preventing, treating, or ameliorating symptoms associated with a disease, disorder, or infection. In any method, therapeutic indication, use or other disclosure herein, the subject may be any animal, typically a mammal, preferably a human.

[0103] Accordingly, the invention provides a method of treating and / or preventing a disease or disorder, the method comprising administering to the subject between about 1 mg to about 15 mg of an AMYR agonist, or pharmaceutically acceptable salt thereof, of the invention. The invention also provides an AMYR agonist, or pharmaceutically acceptable salt thereof, of the invention for use in a method of treating and / or preventing a disease or disorder, the method comprising administering to the subject between about 1 mg to about 15 mg of the AMYR agonist, or pharmaceutically acceptable salt thereof.

[0104] In particular, the invention provides a method of reducing the body weight of a subject, the method comprising administering to the subject between about 1 mg to about 15 mg of an AMYR agonist, or pharmaceutically acceptable salt thereof, of the invention.

[0105] Also provided is an AMYR agonist or a pharmaceutically acceptable salt thereof of the invention for use in a method of reducing the body weight of a subject, the method comprising administering to the subject between about 1 mg to about 15 mg of an AMYR agonist, or pharmaceutically acceptable salt thereof, of the invention.

[0106] Also provided is the use of an AMYR agonist in the manufacture of a medicament for reducing the body weight of a subject, wherein the medicament comprises about 1 mg to about 15 mg of the AMYR agonist.

[0107] Also provided is a method of reducing the body weight of a subject, the method comprising administering to the subject an AMYR agonist, or pharmaceutically acceptable salt thereof, of the invention, wherein the reduction in body weight is fat-specific weight loss. The invention also provides an AMYR agonist, or pharmaceutically acceptable salt thereof, of the invention for use in a method of reducing the body weight of a subject, wherein the reduction in body weight is fat-specific weight loss. The invention also provides the use of an AMYR agonist, or pharmaceutically acceptable salt thereof, of the invention in the manufacture of a medicament for reducing the body weight of a subject, wherein the reduction in body weight is fat-specific weight loss.

[0108] The use or method may comprise administering a therapeutically effective schedule that has less frequent doses of the AMYR agonist than the therapeutically effective dosing schedule of pramlintide.

[0109] In the context of the invention, “reducing the body weight of a subject” can be measured by any suitable means, and may be determined by a reduction in the absolute amount of body weight in a subject e.g., in grams) from baseline, and / or a percent change in body weight from baseline. Treatment according to the present invention may reduce the body weight of a subject by > 5% from baseline, > 6% from baseline, > 7% from baseline^ > 8% from baseline, > 9% from baseline, > 10% from baseline^ > 15% from baseline^ > 20% from baseline, > 25% from baseline, > 30% from baseline, or more. Thus, the subject may have a weight reduction of > 5% from baseline following treatment. The subject may have a weight reduction of > 10% from baseline following treatment. The subject may have a weight reduction of > 15% from baseline following treatment. The subject may have a weight reduction of > 20% from baseline following treatment.

[0110] In some embodiments, the subject may have a greater reduction in body weight as compared to a therapeutically effective dose of a different anti-obesity drug, such a dual amylin-calcitonin receptor agonist (DACRA, e.g., cagrilintide or davalintide) and / or an incretin mimetic drug, such as a GLP-1 R agonist {e.g., semaglutide). Treatment according to the present invention may result in greater reduction of body weight in a subject by > 5%, > 6%, > 7%, > 8%, > 9%, > 10%, > 15%, > 20%, > 25%, > 30%, or more, as compared to a therapeutically effective dose of a different anti-obesity drug, such a dual amylin-calcitonin receptor agonist (DACRA, e.g., cagrilintide or davalintide) and / or an incretin mimetic drug, such as a GLP-1 R agonist e.g., semaglutide).

[0111] The reduction in body weight may be a reduction in fat-specific weight. In the context of the invention, “reducing fat mass-specific body weight” (also referred to as “reducing fat-specific weight) may comprise reducing the absolute amount of fat-specific body weight in a subject {e.g., in grams), and / or reducing the proportion of the fat-specific body weight component of total body weight {e.g., as a percentage of total body weight). A method of the invention may preferentially reduce fat-mass specific body weight as compared to lean mass-specific {e.g. muscle) body weight {e.g., preserving lean-mass specific body weight while reducing overall body weight). For the avoidance of doubt, any and all reference herein to “reducing the body weight of a subject” applies equally and without reservation to “reducing fat massspecific body weight of a subject”, unless expressly stated to the contrary.

[0112] In some embodiments, the subject may have a greater reduction in fat mass-specific body weight as compared to a therapeutically effective dose of a different anti-obesity drug, such a dual amylin-calcitonin receptor agonist (DACRA, e.g., cagrilintide or davalintide) and / or an incretin mimetic drug, such as a GLP-1 R agonist {e.g., semaglutide). Treatment according to the present invention may result in greater reduction of fat mass-specific body weight in a subject by > 5%, > 6%, > 7%, > 8%, > 9%, > 10%, > 15%, > 20%, > 25%, > 30%, or more, as compared to a therapeutically effective dose of a different anti-obesity drug, such a dual amylin-calcitonin receptor agonist (DACRA, e.g., cagrilintide or davalintide) and / or an incretin mimetic drug, such as a GLP-1 R agonist {e.g., semaglutide). The reduction in body weight may be cosmetic (non-therapeutic), e.g. when the patient wishes to lose weight for aesthetic reasons. Thus, the invention provides a cosmetic method of reducing the body weight of a subject, the method comprising administering to the subject between about 1 mg to about 15 mg of an AMYR agonist, or pharmaceutically acceptable salt thereof, of the invention. The invention also provides a cosmetic use of an AMYR agonist, or pharmaceutically acceptable salt thereof, of the invention for reducing the body weight of a subject, wherein said use comprises administering to the subject between about 1 mg to about 15 mg of the AMYR agonist, or pharmaceutically acceptable salt thereof.

[0113] The reduction in body weight may be therapeutic, e.g. when the subject is obese, suffers from an obesity-related condition and / or suffers from a metabolic disease.

[0114] Accordingly, the AMYR agonist may be used in the treatment and / or prevention of a disease or disorder selected from obesity, metabolic disease, an obesity-related condition, eating disorder, Alzheimer's disease, hepatic steatosis ("fatty liver"), kidney failure, arteriosclerosis (e.g. atherosclerosis), cardiovascular disease, macrovascular disease, microvascular disease, diabetic heart (including diabetic cardiomyopathy and heart failure as a diabetic complication), coronary heart disease, peripheral artery disease or stroke, cancer, dumping syndrome, hypertension e.g. pulmonary hypertension, or dyslipidemia e.g. atherogenic dyslipidemia, cholescystitis, or short bowel syndrome. The invention also provides an AMYR agonist for use in such methods.

[0115] In particular, it will be understood that the AMYR agonist may be used in the treatment and / or prevention of obesity, metabolic diseases (such as diabetes, e.g. type 1 or type 2 diabetes), and / or obesity-related conditions. The invention also provides an AMYR agonist for use in such methods. Thus, the subject to be treated may have diabetes {e.g. type 1 or type 2 diabetes).

[0116] Alternatively, the subject may not have type 1 or type 2 diabetes. Alternatively or in addition, the subject may not have obesity induced by other endocrine disorders, such as Cushing’s syndrome or Prader-Willi syndrome. Further alternatively or in addition, the subject may not have previous or planned (within the treatment course) bariatric surgery or fitting of a weight loss device (e.g. a gastric balloon or duodenal barrier). Further alternatively or in addition, the subject may not have gastroparesis.

[0117] The subject may have obesity e.g., a BMI of > 30 kg / m2). Thus, preferably, the AMYR agonist may be used in a method of treating and / or preventing obesity. Said method may comprise administering between about 1 mg to about 15 mg of the AMYR to the subject. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist.

[0118] The subject may have an obesity-related condition. Thus, the AMYR agonist may be used in a method of treating and / or preventing an obesity-related condition in a subject. Said method may comprise administering between about 1 mg to about 15 mg of the AMYR to the subject. The obesity-related condition may be overweight, morbid obesity, obesity prior to surgery, obesity-linked inflammation, obesity-linked gallbladder disease, sleep apnoea and respiratory problems, hyperlipidaemia, degeneration of cartilage, osteoarthritis, or reproductive health complications of obesity or overweight such as infertility in a subject. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist.

[0119] The subject may be overweight e.g., a BMI of > 27 kg / m2) and have at least one weight- related comorbidity e.g., CV disease, hypertension, dyslipidaemia or hyperlidemia, and / or obstructive sleep apnoea). Thus, preferably, the AMYR agonist may be used in a method of treating and / or preventing overweight. Said method may comprise administering between about 1 mg to about 15 mg of the AMYR to the subject. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist.

[0120] The subject may have a metabolic disease. Thus, the AMYR agonist may be used in a method of treating and / or preventing a metabolic disease in a subject. Said method may comprise administering between about 1 mg to about 15 mg of the AMYR to the subject. Said metabolic disease may be diabetes, type 1 diabetes, type 2 diabetes, gestational diabetes, pre-diabetes, insulin resistance, impaired glucose tolerance (IGI), disease states associated with elevated blood glucose levels, metabolic disease including metabolic syndrome, or hyperglycaemia, e.g. abnormal postprandial hyperglycaemia. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist.

[0121] The subject may tolerate the AMYR agonist well. Tolerance of the AMYR agonist may be assessed using any appropriate means, which are within the routine practice of one of ordinary skill in the art. Non-limiting examples of parameters which may be used to assess tolerance include nausea and vomiting. A subject who tolerates the AMYR agonist well may exhibit reduced nausea and / or vomiting (e.g. reduced incidence of nausea and / or vomiting). Alternatively or in addition, tolerance to the AMYR agonist may be assessed by change in growth / differentiation factor 15 (GDF15) from baseline ( / .e. reduced GDF15). Change in GDF15 may be assessed using any suitable sample from the subject, as described herein, e.g. a blood sample. The subject may have increased tolerability to the AMYR agonist (such as reduced nausea and / or vomiting) as compared with their tolerance to a therapeutically effective dose of a different anti-obesity drug, such a dual amylin-calcitonin receptor agonist (DACRA) and / or an incretin mimetic drug, such as a GLP-1 R agonist {e.g., semaglutide). Treatment according to the present invention may reduce the incidence of nausea of a subject by > 5%, > 6%, > 7%, > 8%, > 9%, > 10%, > 15%, > 20%, > 25%, > 30%, or more, as compared to a therapeutically effective dose of a different anti-obesity drug, such a dual amylin-calcitonin receptor agonist (DACRA, e.g., davalintide) and / or an incretin mimetic drug, such as a GLP-1 R agonist e.g., semaglutide). Treatment according to the present invention may reduce the incidence of vomiting of a subject by > 5%, > 6%, > 7%, > 8%, > 9%, > 10%, > 15%, > 20%, > 25%, > 30%, or more, as compared to a therapeutically effective dose of a different anti-obesity drug, such a dual amylin-calcitonin receptor agonist (DACRA, e.g., davalintide) and / or an incretin mimetic drug, such as a GLP-1 R agonist {e.g., semaglutide). Treatment according to the present invention may reduce the levels of GDF- 15 of a subject by > 5%, > 6%, > 7%, > 8%, > 9%, > 10%, > 15%, > 20%, > 25%, > 30%, or more, from baseline.

[0122] Treatment according to the present invention may reduce the need for the subject to be administered an anti-emetic medication as compared to a therapeutically effective dose of a different anti-obesity drug, such a dual amylin-calcitonin receptor agonist (DACRA, e.g., davalintide) and / or an incretin mimetic drug, such as a GLP-1 R agonist (e.g., semaglutide). Suitable anti-emetic medications will be familiar to a person skilled in the art. Thus, treatment according to the present invention may allow the subject to be administered a reduced dose of an anti-emetic medication as compared to a therapeutically effective dose of a different anti-obesity drug. Alternatively or in addition, treatment according to the present invention may allow the subject to be administered an anti-emetic medication at a reduced dosing frequency as compared to a therapeutically effective dose of a different anti-obesity drug. Treatment according to the present invention may eliminate the need for a subject to be administered an anti-emetic medication, i.e., the subject is not administered an anti-emetic medication.

[0123] Also provided is a method of inhibiting or reducing weight gain, promoting weight loss, reducing food intake, increasing satiety, and / or reducing excess body weight, the method comprising about 1 mg to about 15 mg of the AMYR agonist of the invention to the subject. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. Said method may be therapeutic, e.g. when the subject is obese, suffers from an obesity- related condition and / or a metabolic disease. Alternatively, said method may be cosmetic (non-therapeutic), e.g. when the patient wishes to lose weight for aesthetic reasons.

[0124] Thus, the invention also provides a cosmetic use of about 1 mg to about 15 mg of the AMYR agonist of the invention to inhibit or reduce weight gain, promote weight loss, reduce food intake, increase satiety, and / or reduce excess body weight in a subject.

[0125] In some embodiments, the subject may have a greater reduction in food intake as compared to a therapeutically effective dose of a different anti-obesity drug, such a dual amylin- calcitonin receptor agonist (DACRA, e.g., cagrilintide or davalintide) and / or an incretin mimetic drug, such as a GLP-1 R agonist {e.g., semaglutide). Treatment according to the present invention may result in greater reduction of food intake in a subject by > 5%, > 6%, > 7%s> 8%, > 9%, > 10%3> 15%s> 20%s> 25%, > 30%, or more, as compared to a therapeutically effective dose of a different anti-obesity drug, such a dual amylin-calcitonin receptor agonist (DACRA, e.g., cagrilintide or davalintide) and / or an incretin mimetic drug, such as a GLP-1 R agonist e.g., semaglutide).

[0126] There is also provided a method of reducing fat mass-specific body weight (as defined herein), the method comprising administering about 1 mg to about 15 mg of the AMYR agonist of the invention to the subject. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. Said method may be therapeutic, e.g. when the subject is obese, suffers from an obesity-related condition and / or a metabolic disease. Alternatively, said method may be cosmetic (non-therapeutic), e.g. when the patient wishes to lose weight for aesthetic reasons.

[0127] Thus, the invention also provides a cosmetic use of about 1 mg to about 15 mg of the AMYR agonist of the invention to reduce fat-mass specific body weight in a subject.

[0128] There is also provided a method of improving body composition, the method comprising administering about 1 mg to about 15 mg of the AMYR agonist of the invention to the subject. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. The skilled person will be familiar with a suitable method for assessing body composition, for example, absolute change and / or percentage change (e.g., a reduction) in BMI, total body mass, body fat, muscle mass, and / or fat free mass (in kg). By way of non-limiting example, an improvement in body composition may be determined by a reduction in waist circumference (in cm) from baseline. By way of another non-limiting example, an improvement in body composition may be determined by bioimpedance. Said method may be therapeutic, e.g. when the subject is obese, suffers from an obesity-related condition and / or a metabolic disease. Alternatively, said method may be cosmetic (non-therapeutic), e.g. when the patient wishes to improve body composition for aesthetic reasons.

[0129] Thus, the invention also provides a cosmetic use of about 1 mg to about 15 mg of the AMYR agonist of the invention to improve body composition in a subject.

[0130] There is also provided a method of reducing the waist circumference in a subject, the method comprising administering about 1 mg to about 15 mg of the AMYR agonist of the invention to the subject. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. The skilled person will be familiar with a suitable method for measuring waist circumference. By way of non-limiting example, waist circumference may be defined as the minimal abdominal circumference located midway between the lower rib margin and the iliac crest. Measurement (in cm) may be performed with the subject in a standing position with arms down their side and feet together while breathing normally. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. Said method may be therapeutic, e.g. when the subject is obese, suffers from an obesity-related condition and / or a metabolic disease. Alternatively, said method may be cosmetic (non-therapeutic), e.g. when the patient wishes to lose weight for aesthetic reasons.

[0131] Thus, the invention also provides a cosmetic use of about 1 mg to about 15 mg of the AMYR agonist of the invention to reduce the waist circumference in a subject.

[0132] The invention also provides a method of improving glycemic and / or metabolic control in a subject, the method comprising administering about 1 mg to about 15 mg of the AMYR agonist of the invention to the subject.

[0133] The method of improving glycemic and / or metabolic control in a subject may comprise or consist of increasing insulin secretion, delaying gastric emptying, increasing mitochondria function, inhibiting de novo lipogenesis, decreasing HbAlc, enhancing fatty oxidation, decreasing hepatic mitochondrial oxidative stress, decreasing steatosis, decreasing fibrosis, decreasing glycogen synthesis, increasing gluconeogenesis, reducing or reversing fibrosis e.g., liver fibrosis), reducing steatohepatitis, and / or reducing risk of death due to cirrhosis, hepatocellular carcinoma, and / or cardiorenal disease in the subject.

[0134] There is also provided a method of improving glucose homeostasis, p-cell function, and / or insulin sensitivity in a subject, the method comprising administering about 1 mg to about 15 mg of the AMYR agonist of the invention to the subject. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. An improvement in glucose homeostasis, P-cell function, and / or insulin sensitivity may be measured by change from baseline in the updated homeostatic model assessment (H0MA2) scores (HOMA2-IR, HOMA2-%B, and HOMA2-%S based on glucose and C-peptide) over time. H0MA2 is well-known in the art, such as described in Levy et al. (Diabetes Care (1998) 21 (12):2191 -2192), which is herein incorporated by reference in its entirety. An improvement in glucose homeostasis, p-cell function, and / or insulin sensitivity may be measured by change from baseline in HbA1c, fasting serum glucose, fasting serum insulin, fasting serum c-peptide, and / or plasma proinsulin levels over time. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist.

[0135] Also provided is a method of promoting end-organ protection e.g., cardiovascular, renal and / or liver) in a subject, the method comprising administering about 1 mg to about 15 mg of the AMYR agonist of the invention to the subject. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. End-organ protection may be measured by assessing change from baseline on cardiovascular endpoints, such as ApoB / ApoA1 and / or high sensitivity Troponin T (hs Troponin T). Alternatively or in addition, end-organ protection can be measured by assessing change on renal endpoints, such as estimated glomerular filtration rate (eGFR, calculated using creatinine and cystatin C), UACR (urine albumin and creatinine ratio), and / or renin. Further alternatively or in addition, end-organ protection can be measured by assessing change from baseline on liver endpoints, such as alanine transaminase (ALT), aspartate aminotransferase (AST), and / or FIB-4 (which is a standard clinical index for measuring liver fibrosis, calculated using AST, ALT, age, and / or platelet). The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. Said method may be therapeutic, e.g. when the subject is obese, suffers from an obesity - related condition and / or a metabolic disease. Alternatively, said method may be cosmetic (non-therapeutic), e.g. when the patient wishes to lose weight for aesthetic reasons.

[0136] Also provided is a method of improving weight-loss quality in a subject, the method comprising administering about 1 mg to about 15 mg of the AMYR agonist of the invention to the subject. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. Weight loss quality may be measured by change from baseline to on metabolic / adipokine health, as assessed by levels of adiponectin and / or leptin, lipid profile (such as levels of triglycerides (TG), total cholesterol, non-esterified fatty acids (NEFA), high- density lipoprotein (HDL), low-density lipoprotein (LDL), and / or very low-density lipoprotein (VLDL)), and / or change from baseline in systemic inflammation (such as levels of C-reactive protein (CRP), e.g. a high-sensitivity CRP test (hsCRP), and / or interleukin-6 (IL-6), e.g. a high-sensitivity IL-6 test (hslL6)). The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. Said method may be therapeutic, e.g. when the subject is obese, suffers from an obesity-related condition and / or a metabolic disease. Alternatively, said method may be cosmetic (non-therapeutic), e.g. when the patient wishes to lose weight for aesthetic reasons.

[0137] Thus, the invention also provides a cosmetic use of about 1 mg to about 15 mg of the AMYR agonist of the invention to improve weight-loss quality in a subject.

[0138] Also provided is a method of improving physical function in a subject, the method comprising administering about 1 mg to about 15 mg of the AMYR agonist of the invention to the subject. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. Improving physical function in a subject may be assessed by change from baseline in IWQoL-Lite CT Physical Function score, SF-36 v2 Physical functioning score, IWQoL-Lite CT Total Score, SF-36 v2 physical component, mental component scores, and / or FACIT- GP5 scores.

[0139] Also provided herein is about 1 mg to about 15 mg of the AMYR agonist of the invention for use in the manufacture of a medicament. Said medicament may be for use in the treatment and / or prevention of any disease or disorder, or for any clinical purpose (e.g. improving body composition, improving glycemic and / or metabolic control, improving glucose homeostasis, P-cell function, and / or insulin sensitivity, or promoting end-organ protection) as described herein.

[0140] The route of administration of an AMYR agonist, or pharmaceutical composition thereof, can be, for example, oral, parenteral, by inhalation or topical. Preferably, the AMYR agonist or pharmaceutical composition thereof is administered by parenteral administration to a subject or patient. The term “parenteral” as used herein includes, e.g., intravenous, intraarterial, intraperitoneal, intramuscular, subcutaneous, rectal, or vaginal administration. Preferably, the AMYR agonist or pharmaceutical composition thereof is administered by injection, such as by intravenous, subcutaneous or intramuscular injection, to a subject or patient. In particularly preferred embodiments, the AMYR agonist or pharmaceutical composition thereof is administered by subcutaneous injection. Administration by injection, such as by subcutaneous injection, offers the advantage of better comfort for the subject or patient and the opportunity to administer to a subject or patient outside of a hospital setting. Accordingly, the AMYR agonist or pharmaceutical composition thereof may be administered by selfadministration.

[0141] The subject or patient may be a mammal, particularly a human.

[0142] The AMYR agonist or pharmaceutical composition thereof may be administered to the subject in combination with insulin and / or another anti-obesity drug, such as a leptin.

[0143] Any AMYR agonist may be used for therapy, in a method of treatment or in the manufacture of a medicament according to the present invention. Preferably, any AMYR agonist which has selectivity for AMYR as compared with CTR may be used for therapy, in a method of treatment or in the manufacture of a medicament according to the present invention. Preferably, the AMYR agonist with selectivity for AMYR as compared with CTR is an AMYR agonist polypeptide, as described herein.

[0144] The subject to be treated may be a human having a BMI of at least 25 kg / m2at baseline.

[0145] The subject to be treated may be a human having a BMI of at least 27 kg / m2at baseline.

[0146] The subject to be treated may be a human having a BMI of at least 30 kg / m2. The subject may be a human having a BMI of at least 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37,

[0147] 38, 39 or 40 kg / m2at baseline. The subject may be a human having a BMI of less than or equal to 50 kg / m2at baseline. The subject may be a human having a BMI of less than or equal to 50, 49, 48, 47, 46, 45, 44, 43, 42, 41 , or 40 kg / m2at baseline. The subject may be a human having a BMI of 25-40 kg / m2at baseline. The subject may be a human having a BMI of 27-40 kg / m2at baseline. The subject may be a human having a BMI of 30-40 kg / m2at baseline. Preferably, the subject is a human having a BMI of at least 30 kg / m2at baseline. Preferably, the subject is a human having a BMI of at least 27 kg / m2at baseline with at least one weight-related comorbidity e.g., CV disease, hypertension, dyslipidaemia or hyperlipidaemia, and / or obstructive sleep apnoea).

[0148] The subject may have previously been administered an anti-obesity drug, such as a dual amylin and calcitonin receptor agonist (DACRA) or an incretin mimetic drug, such as a GLP- 1 R agonist e.g., semaglutide). In some embodiments, the subject does not tolerate an antiobesity drug, such as a DACRA or an incretin mimetic drug, such as a GLP-1 R agonist e.g., semaglutide). As used herein, a subject that does not or cannot tolerate a drug may have had, or be at risk of, adverse events (such as nausea and / or vomiting) in response to the drug. Thus, the subject may have experienced adverse events following administration of an anti-obesity drug; discontinued administration of the anti-obesity drug due to the burden of adverse events of the drug; or been advised that they are at risk of adverse events if administered an anti-obesity drug.

[0149] Dosage regimens

[0150] The invention provides a method of reducing the body weight of a subject, the method comprising administering to the subject about 1 mg to about 15 mg of an amylin receptor (AMYR) agonist. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. The invention also provides the use of an AMYR agonist in the manufacture of a medicament for use in such a method, the method comprising administering to the subject the medicament which comprises about 1 mg to about 15 mg of the AMYR agonist.

[0151] The invention also provides a method of treating and / or preventing disease or disorder as disclosed herein, the method comprising administering to a subject in need thereof about 1 mg to about 15 mg of the AMYR agonist. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. The invention also provides an AMYR agonist in the manufacture of a medicament for treating and / or preventing a disease or disorder as disclosed herein, wherein the medicament comprises about 1 mg to about 15 mg of the AMYR agonist. In particular, the invention provides a method of treating and / or preventing obesity or an obesity-related condition in a subject in need thereof, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. The invention also provides an AMYR agonist in the manufacture of a medicament for treating and / or preventing obesity or an obesity-related condition as disclosed herein, wherein the medicament comprises about 1 mg to about 15 mg of the AMYR agonist.

[0152] The invention also provides a method of treating and / or preventing a metabolic disease in a subject in need thereof, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. The invention also provides an AMYR agonist for use in such a method, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist. The invention also provides an AMYR agonist in the manufacture of a medicament for treating and / or preventing a metabolic disease as disclosed herein, wherein the medicament comprises about 1 mg to about 15 mg of the AMYR agonist.

[0153] Also provided is a kit comprising (a) an AMYR agonist, and (b) instructions for use of the same for treating or preventing a disease or disorder as disclosed herein (such as obesity or an obesity-related condition, or a metabolic disease as described herein) at a dose of about 1 mg to about 15 mg.

[0154] The amylin receptor agonist in a method of the invention, an amylin receptor agonist for use in the invention, and the uses, compositions or kits of the invention may be any amylin receptor agonist as disclosed herein.

[0155] Preferably, the doses as disclosed herein are fixed doses. As used herein, the term "fixed dose" refers to a dose which is used for all subjects, e.g., the dose is an amount that does not vary based on the weight of the subject. A fixed dose may be a specific, unchanging amount, wherein the same dose is used for all subjects, or for all adult subjects, such as a unit dosage form as disclosed herein. A fixed dose is typically expressed in terms of an amount of the active agent, such as the amylin receptor agonist or pharmaceutically acceptable salt thereof, rather than an amount that is expressed relative to the weight or mass of the subject to be treated. The methods, uses, compositions, and kits of the invention utilise a dose (such as a fixed dose) of about 1 mg to about 15 mg of an AMYR agonist, or pharmaceutically acceptable salt thereof, for example comprise administering such a (fixed) dose to a subject.

[0156] The (fixed) dose of about 1 mg to about 15 mg may be at least 1 mg, such as at least 2 mg, at least 3 mg, at least 4 mg, at least 5 mg, at least 6 mg, at least 7 mg, at least 8 mg, at least

[0157] 9 mg, at least 10 mg, at least 11 mg, or at least 12 mg. The (fixed) dose of about 1 mg to about 15 mg may be up to 15 mg, such as up to 12 mg, up to 11 mg, up to 10 mg, up to 9 mg, up to 8 mg, up to 7 mg, up to 6 mg, up to 5 mg, up to 4 mg, up to 3 mg, or up to 2 mg. Any of these upper and lower end points may be combined within the range of about 1 mg to about 15 mg. For example, the AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about

[0158] 10 mg, about 1 mg to about 6 mg, about 1 mg to about 5 mg, about 4 mg to about 10 mg, about 1 mg to about 3 mg, about 2.5 mg to about 5 mg, or about 7 mg to about 10 mg.

[0159] The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 12 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 10 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 1 .5 mg to about 9 mg.

[0160] The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 8 mg, about 1 mg to about 7 mg, about 1 mg to about 6 mg, or about 1 mg to about 5 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 1 .5 mg to about 8 mg, about 1 .5 mg to about 7 mg, about 1 .5 mg to about 6 mg, about 1.5 mg to about 5 mg, or about 1.5 mg to about 4.5 mg. Preferably, the AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 5 mg. Preferably, the AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 1 .5 mg to about 4.5 mg.

[0161] The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 1.6 mg to about 8 mg, about 1.6 mg to about 7 mg, about 1.6 mg to about 6 mg, about 1.6 mg to about 5 mg, or about 1.6 mg to about 4.5 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose of about 2 mg to about 15 mg, about 3 mg to about 15 mg, about 4 mg to about 15 mg, about 4.5 mg to about 15 mg, or about 6 mg to about 15 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose of about 2 mg to about 12 mg, about 3 mg to about 12 mg, about 4 mg to about 12 mg, about 4.5 mg to about 12 mg, or about 6 mg to about 12 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose of about 2 mg to about 10 mg, about 3 mg to about 10 mg, about 4 mg to about 10 mg, about 4.5 mg to about 10 mg, or about 6 mg to about 10 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose of about 2 mg to about 9 mg, about 3 mg to about 9 mg, about 4 mg to about 9 mg, about 4.5 mg to about 9 mg, or about 6 mg to about 9 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 4 mg to about 15 mg, about 4 mg to about 12 mg, about 4 mg to about 10 mg, about 4 mg to about 9 mg, or about 6 mg to about 9 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 6 mg to about 15 mg, about 6 mg to about 12 mg, about 6 mg to about 10 mg, or about 6 mg to about 9 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 4.5 mg to about 15 mg, about 4.5 mg to about 12 mg, about 4.5 mg to about 10 mg, or about 4.5 mg to about 9 mg. Preferably, the AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 4 mg to about 10 mg. Preferably, the AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 4 mg to about 9 mg. Preferably, the AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 4.5 mg to about 9 mg.

[0162] The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose of about 1 mg to about 4 mg, about 1 mg to about 3 mg, or about 1 mg to about 2 mg. Preferably, the AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 3 mg.

[0163] The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 2 mg to about 8 mg, about 2 mg to about 7 mg, about 2 mg to about 6 mg, or about 2 mg to about 5 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 3 mg to about 8 mg, about 3 mg to about 7 mg, about 3 mg to about 6 mg, or about 3 mg to about 5 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 4 mg to about 8 mg, about 4 mg to about 7 mg, about 4 mg to about 6 mg, or about 4 mg to about 5 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 2.5 mg to about 8 mg, about 3 mg to about 8 mg, or about 4 mg to about 8 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 2.5 mg to about 7 mg, about 3 mg to about 7 mg, or about 4 mg to about 7 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 2.5 mg to about 6 mg, about 3 mg to about 6 mg, or about 4 mg to about 6 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 2.5 mg to about 5 mg, about 3 mg to about 5 mg, or about 4 mg to about 5 mg. Preferably, the AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 2.5 mg to about 5 mg.

[0164] The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 5 mg to about 12 mg, about 5 mg to about 10 mg, or about 5 mg to about 9 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 6 mg to about 12 mg, about 6 mg to about 10 mg, or about 6 mg to about 9 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 7 mg to about 12 mg, about 7 mg to about 10 mg, or about 7 mg to about 9 mg. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 8 mg to about 9 mg. Preferably, the AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, of about 7 mg to about 10 mg.

[0165] The AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of up to 15 mg. For example, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 10 mg, about 1 mg to about 5 mg, about 4 mg to about 10 mg, about 1 mg to about 3 mg, about 2.5 mg to about 5 mg, or about 7 mg to about 10 mg. The AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of up to 12 mg. For example, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 10 mg, about 1 mg to about 5 mg, about 4 mg to about 10 mg, about 1 mg to about 3 mg, about 2.5 mg to about 5 mg, or about 7 mg to about 10 mg.

[0166] The AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of up to 10 mg. For example, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 10 mg, about 1 mg to about 5 mg, about 4 mg to about 10 mg, about 1 mg to about 3 mg, about 2.5 mg to about 5 mg, or about 7 mg to about 10 mg.

[0167] The AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of up to 9 mg. For example, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 9 mg, about 1 mg to about 5 mg, about 4 mg to about 9 mg, about 1 mg to about 3 mg, about 2.5 mg to about 5 mg, about 7 mg to about 9 mg, or about 9 mg.

[0168] The AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of up to 8 mg. For example, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 8 mg, about 1 mg to about 5 mg, about 4 mg to about 8 mg, about 1 mg to about 3 mg, about 2.5 mg to about 5 mg, or about 7 mg to about 8 mg.

[0169] The AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of up to 7 mg. For example, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 7 mg, about 1 mg to about 5 mg, about 4 mg to about 7 mg, about 1 mg to about 3 mg, or about 2.5 mg to about 5 mg.

[0170] The AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of up to 6 mg. For example, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 6 mg, about 1 mg to about 5 mg, about 4 mg to about 6 mg, about 1 mg to about 3 mg, or about 2.5 mg to about 5 mg.

[0171] The AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of up to 5 mg. For example, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 5 mg, about 4 mg to about 5 mg, about 1 mg to about 3 mg, about 2.5 mg to about 5 mg, or about 4.5 mg.

[0172] The AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of up to 4 mg. For example, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 4 mg, about 1 mg to about 3 mg, or about 2.5 mg to about 4 mg.

[0173] The AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of up to 3 mg. For example, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 3 mg.

[0174] The AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of up to 2 mg. For example, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 1 mg to about 2 mg, about 1 .5 mg, or about 1 .6 mg.

[0175] Preferably, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 1 .5 mg, about 1 .6 mg, about 4.5 mg, about 6 mg, or about 9 mg.

[0176] Preferably, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 1 .5 mg, about 4.5 mg, or about 9 mg.

[0177] Preferably, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 1 .5 mg, such as a dose of 1 .5 mg.

[0178] Preferably, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 1 .6 mg, such as a dose of 1 .6 mg.

[0179] Preferably, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 4.5 mg, such as a dose of 4.5 mg.

[0180] Preferably, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 6 mg, such as a dose of 6 mg.

[0181] Preferably, the AMYR agonist may be administered to the subject at a dose, such as a fixed dose, of about 9 mg, such as a dose of 9 mg. Treatment or prevention may be effected by a single administration or by multiple administrations of the AMYR agonist. For example, the treatment or prevention may comprise administering two doses, three doses, four doses, five doses, six doses, seven doses, eight doses, nine doses, ten doses, or more than ten doses of the AMYR agonist to the subject. Each dose may be a dose, such as a fixed dose, as disclosed herein. The treatment or prevention may comprise multiple administrations as described herein, such as repeated administrations, of the AMYR agonist over a course of treatment.

[0182] By a course of treatment is meant a treatment plan comprising several rounds of administration, as part of a method of reducing body weight or treating and / or preventing a disease or disorder as described herein (e.g. obesity or an obesity-related condition and / or a metabolic disease) in accordance with the invention. A course of treatment may last for one or more weeks, one or more months, such as two, three, four, five, six, seven, eight, nine, ten or eleven months, or for one or more years, such as one year, two years, three years, four years, five years, or longer. A course of treatment may be continued with no fixed endpoint, for example a course of treatment may be continued for the lifetime of the subject being treated. Multiple administrations, such as repeated administrations, may be continued at regular or irregular intervals during the course of the treatment as determined to be needed by a medical practitioner. A course of treatment of any duration as described herein may comprise a titration schedule as discussed below.

[0183] The AMYR agonist or pharmaceutical composition thereof may be administered to a subject multiple times, at an interval of once a week, every two weeks, every three weeks, or every four weeks. The AMYR agonist or pharmaceutical composition thereof may be administered about once every 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, or 14 days. Preferably, the AMYR agonist or pharmaceutical composition thereof is administered about once a week. Accordingly, the AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, in the range of about 1 mg to about 10 mg, about 1 mg to about 5 mg, about 4 mg to about 10 mg, about 1 mg to about 3 mg, about 2.5 mg to about 5 mg, or about 7 mg to about 10 mg about once a week. The method may comprise administering to the subject a dose of about 1 .5 mg, about 4.5 mg, or about 9 mg about once a week.

[0184] When the AMYR agonist is administered in multiple administrations, such as repeated administrations, each of said administrations may be at a fixed dose as described herein. For example, each administration in a method or use of the invention may comprise a fixed dose of about 1 mg to about 15 mg of the AMYR agonist as disclosed herein. In such instances, the number of administrations or the frequency of administrations may be any of the options disclosed herein. The multiple administrations, such as repeated administrations of the AMYR agonist, may be each at the same dose. Two or more administrations of the AMYR agonist may be each at the same dose. The multiple administrations, such as repeated administrations of the AMYR agonist, may be each at a dose within a range of doses disclosed herein. For example, the AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, in the range of about 1 mg to about 10 mg, about 1 mg to about 5 mg, about 4 mg to about 10 mg, about 1 mg to about 3 mg, about 2.5 mg to about 5 mg, or about 7 mg to about 10 mg. The method may comprise administering to the subject multiple doses of about 1.5 mg, about 1.6 mg, about 4.5 mg, about 6 mg or about 9 mg. The method may comprise administering to the subject multiple doses of about 1 .5 mg, about 4.5 mg, or about 9 mg.

[0185] In accordance with the invention, the AMYR agonist may be administered to a subject multiple times over a period of at least 1 month, 2 months, 3 months, 6 months, 9 months, 1 year, 2 years or 5 years. The AMYR agonist may be administered to a subject about once a week over a period of at least 1 month, 2 months, 3 months, 6 months, 9 months, 1 year, 2 years or 5 years. The subject may have body weight reduction following administration of multiple doses after 3 months. The subject may have body weight reduction following administration of multiple doses after 6 months. The subject may have body weight reduction following administration of multiple doses between a period of about 3 months and about 6 months. The AMYR agonist may be administered at a dosing interval of about once a week over these periods of time.

[0186] For example, the AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, in the range of about 1 mg to about 10 mg, about 1 mg to about 5 mg, about 4 mg to about 10 mg, about 1 mg to about 3 mg, about 2.5 mg to about 5 mg, or about 7 mg to about 10 mg about once a week over a period of at least 3 months. The method may comprise administering to the subject a dose of about 1.5 mg, about 1.6 mg, about 4.5 mg, about 6 mg, or about 9 mg about once a week over a period of at least 3 months. The method may comprise administering to the subject a dose of about 1.5 mg, about 4.5 mg, or about 9 mg about once a week over a period of at least 3 months. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, in the range of about 1 mg to about 10 mg, about 1 mg to about 5 mg, about 4 mg to about 10 mg, about 1 mg to about 3 mg, about 2.5 mg to about 5 mg, or about 7 mg to about 10 mg about once a week over a period of at least 6 months. The method may comprise administering to the subject a dose of about 1.5 mg, about 1.6 mg, about 4.5 mg, about 6 mg, or about 9 mg about once a week over a period of at least 6 months. The method may comprise administering to the subject a dose of about 1.5 mg, about 4.5 mg, or about 9 mg about once a week over a period of at least 6 months. The AMYR agonist, or pharmaceutically acceptable salt thereof, may be administered to the subject at a dose, such as a fixed dose, in the range of about 1 mg to about 10 mg, about 1 mg to about 5 mg, about 4 mg to about 10 mg, about 1 mg to about 3 mg, about 2.5 mg to about 5 mg, or about 7 mg to about 10 mg about once a week over a period of between about 3 months to about 6 months. The method may comprise administering to the subject a dose of about 1.5 mg, about 1.6 mg, about 4.5 mg, about 6 mg, or about 9 mg about once a week over a period of at about 3 months to about 6 months. The method may comprise administering to the subject a dose of about 1 .5 mg, about 4.5 mg, or about 9 mg about once a week over a period of about 3 months to about 6 months. By way of non-limiting example, the method may comprise administering to the subject a dose of about 1 .5 mg about once a week over a period of about 3 months to about 6 months (e.g. about 3 months, about 4 months, about 5 months or about 6 months). By way of a further non-limiting example, the method may comprise administering to the subject a dose of about 1 .6 mg about once a week over a period of about 3 months to about 6 months (e.g. about 3 months, about 4 months, about 5 months or about 6 months). By way of a further non-limiting example, the method may comprise administering to the subject a dose of about 4.5 mg about once a week over a period of about 3 months to about 6 months (e.g. about 3 months, about 4 months, about 5 months or about 6 months). By way of a further nonlimiting example, the method may comprise administering to the subject a dose of about 6 mg about once a week over a period of about 3 months to about 6 months (e.g. about 3 months, about 4 months, about 5 months or about 6 months). By way of a further nonlimiting example, the method may comprise administering to the subject a dose of about 9 mg about once a week over a period of about 3 months to about 6 months (e.g. about 3 months, about 4 months, about 5 months or about 6 months).

[0187] The AMYR agonist may be titrated to a final dose over a period of time. The AMYR agonist may be titrated to a final dose over a period between about 1 week to about 8 weeks. The AMYR agonist may be titrated to a final dose over a period between about 2 weeks to about 8 weeks. The AMYR agonist may be titrated to a final dose over a period between about 2 weeks to about 6 weeks. The AMYR agonist may be titrated to a final dose over a period between about 4 weeks to about 8 weeks. The AMYR agonist may be titrated to a final dose over about 2 weeks. The AMYR agonist may be titrated to a final dose over about 4 weeks. The AMYR agonist may be titrated to a final dose over about 6 weeks. The AMYR agonist may be titrated to a final dose over about 8 weeks. Where the AMYR agonist is a dose of about 4.5 mg, the AMYR agonist may be titrated to a dose of about 4.5 mg over about 4 weeks. Where the AMYR agonist is a dose of about 9 mg, the AMYR agonist may be titrated to a dose of about 9 mg over about 4 weeks or over about 6 weeks. Where the AMYR agonist is a dose of about 9 mg, the AMYR agonist may be titrated to a dose of about 9 mg over about 6 weeks. Where the AMYR agonist is a dose of about 9 mg, the AMYR agonist may be titrated to a dose of about 9 mg over about 8 weeks. Titration may involve a single step-change in the dose of the AMYR agonist. Each step-change may comprise one more administrations of a dose (e.g. 1 , 2, 3, or 4 doses) at a given titration level. By way of nonlimiting example, titration to a final dose of 4.5 mg may comprise one or more dose (e.g. 1 , 2, 3, or 4 doses) of 1.5 mg followed by the remaining doses at the final dose of 4.5 mg. Alternatively, titration may involve multiple step-changes (e.g. 2, 3 or more step-changes) in the dose of the AMYR agonist. Each step-change may comprise one more administrations of a dose (e.g. 1 , 2, 3, or 4 doses) at a given titration level. Thus, the AMYR agonist may be titrated at a first dose over a first period of time, increased to one or more intermediate doses over a second or subsequent e.g., a third, fourth, or fifth, etc.) period of time, and then increased to a final dose. The first, second, and subsequent periods of time may be the same or different lengths of time. The first period of time may be between about 1 week to about 8 weeks The first period of time may be between about 2 weeks to about 8 weeks. The first period of time may be between about 2 weeks to about 6 weeks. The first period of time may be between about 4 weeks to about 8 weeks. The first period of time may be about 1 week. The first period of time may be about 2 weeks. The first period of time may be about 4 weeks. The first period of time may be about 6 weeks. The first period of time may be about 8 weeks. The second and / or subsequent period of time may be between about 2 weeks to about 8 weeks. The second and / or subsequent period of time may be between about 2 weeks to about 6 weeks. The second and / or subsequent period of time may be between about 4 weeks to about 8 weeks. The second and / or subsequent period of time may be about 2 weeks. The second and / or subsequent period of time may be about 4 weeks. The second and / or subsequent period of time may be about 6 weeks. The second and / or subsequent period of time may be about 8 weeks. By way of non-limiting example, titration to a final dose of 9 mg may comprise one or more dose (e.g. 1 , 2, 3, or 4 doses) of 1.5 mg, followed by one or more dose (e.g. 1 , 2, 3 or 4 doses) at 4.5 mg, followed by the remaining doses at the final dose of 9 mg. Further exemplary titration schedules are set out in Table 1 herein.

[0188] Amylin Receptor Agonists

[0189] Efficacy Typically the AMYR is human AYMR (hAMYR). Human AMYR is formed of the human calcitonin receptor hCTR complexed with at least one of the human receptor activity modifying proteins designated hRAMPI , hRAMP2 and hRAMP3. Thus, hAMYI R is a complex of hCTR and hRAMPI ; hAMY2R is a complex of hCTR and hRAMP2; and hAMY3R is a complex of hCTR and hRAMP3. An exemplary amino acid sequence for hRAMPI is given in SEQ ID NO: 1. An exemplary amino acid sequence for hRAMP2 is given in SEQ ID NO: 2. An exemplary amino acid sequence for hRAMP3 is given in SEQ ID NO: 3. An exemplary amino acid sequence for hCTR is given in SEQ ID NO: 4.

[0190] An AMYR agonist of the invention may activate one or more of hAMYI R, hAMY2R and / or hAMY3R. Thus, an AMYR agonist of the invention may activate hAMYI R; hAMY2R; hAMY3R; hhAMYI R and AMY2R; hAMYI R and AhMY3R; hAMY2R and hAMY3R; or hAMYI R, hAMY2R and hAMY3R.

[0191] An AMYR agonist is not native amylin but exhibits activity at the AMYR of about at least 1% or more relative to native amylin. In some embodiments, an AMYR agonist exhibits activity at the AMYR of at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more relative to native amylin. This may be measured using a cAMP assay, and quantified by Effective Concentration (EC) values, such as asymptotic maximum attainable response, Emax, or EC5O, as described further below.

[0192] The ability to induce cAMP formation as a result of binding to the relevant receptor or receptor complex is typically regarded as indicative of agonist activity. Other intracellular signaling pathways or events may also be used as readouts for amylin receptor agonist activity. These may include calcium release, arrestin recruitment, receptor internalization, kinase activation or inactivation, lipase activation, inositol phosphate release, diacylglycerol release or nuclear transcription factor translocation.

[0193] EC5O values may be used as a measure of agonist potency at a given receptor. An EC5o value is a measure of the concentration of a compound required to achieve half of that compound's maximal activity in a particular assay, for example a cAMP assay as described in Example 2 of WO 2022 / 129254, which is herein incorporated by reference in its entirety.

[0194] A drug that can agonise both AMYR and CTR with no particular selectivity for either AMYR or CTR is also known as a dual amylin calcitonin receptor agonist (DACRA). An example of a DACRA is davalintide. However, increased CTR engagement is associated with nausea, e.g., as demonstrated in rat aversion studies (Boccia et al. 2022, Mol Metab. 2022 Apr; 58: 101444). In this regard, Phase lib trials of davalintide were stopped as body weight reduction was found to not be superior to an AMYR selective agonist (pramlintide) and due to high incidence of adverse events including nausea.

[0195] Thus, in particular, an AMYR agonist of the invention has selectivity to AMYR (preferably hAMYR) over a CTR (preferably hCTR). Such amylin agonists are described as selective amylin receptor agonists (SARAs). Thus, in some embodiments, the AMYR agonist of the invention is a SARA. In some embodiments, the AMYR agonist of the invention is not a dual amylin and calcitonin receptor agonist (DACRA). A SARA may have selectivity for AMYR over a CTR as described further below in the context of AMYR agonists of the invention.

[0196] An AMYR agonist of the invention may exhibit greater or similar selectivity to hAMYR over hCTR as pramlintide, optionally as measured using cAMP release from binding to hAMYR and hCTR. Pramlintide exhibits at least 10-fold selectivity to hAMYR as compared to hCTR.

[0197] An AMYR agonist of the invention may exhibit a lower or similar selectivity to hAMYR over hCTR as pramlintide, optionally as measured using cAMP release from binding to hAMYR and hCTR. Where an AMYR agonist of the invention exhibits a lower selectivity to hAMYR over hCTR compared with pramlintide, the AMYR agonist of the invention is still selective for hAMYR over hCTR. Pramlintide exhibits at least 10-fold selectivity to hAMYR as compared to hCTR.

[0198] An AMYR agonist of the invention may have at least a 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 12-fold, at least 15-fold, at least 17-fold, at least 20-fold, or at least 25-fold, at least about 50-fold, at least about 75-fold, or at least about 100-fold selectivity to hAMYR over hCTR. In preferred embodiments, the AMYR agonist has at least a 5-fold selectivity to AMYR e.g. hAMYR) over CTR (e.g. hCTR). In preferred embodiments, the AMYR agonist has at least a 10-fold selectivity to AMYR (e.g. hAMYR) over CTR (e.g. hCTR).

[0199] In some embodiments, an AMYR agonist of the invention has around 12-20-fold, around 14- 18-fold, optionally around 16-fold selectivity to AMYR (e.g., hAMYR) over CTR (e.g., hCTR).

[0200] In some embodiments, the AMYR agonist of the invention has an EC50 measured under the conditions described in Example 2 of WO 2022 / 129254 (i.e. containing 0.1 % bovine serum albumin (BSA)) of below about 1 .4 nM, below about 1 .2 nM, below about 1 nM, below about 0.8 nM, below about 0.6 nM, below about 0.4 nM, below about 0.3 nM, or below about 0.2 nM for an AMYR, particularly AMY3R.

[0201] In contrast, a DACRA may have less than 2-fold selectivity, less than 1.5-fold selectivity, or less than 1.2-fold selectivity for hAMYR over hCTR. A DACRA may have approximately equal selectivity for hAMYR and hCTR. A DACRA may have at least a 1 .2-fold, at least a 1 .5-fold, or at least a 2-fold selectivity to hCTR over hAMYR.

[0202] Classification

[0203] The AMYR agonist of the invention may be a polypeptide, a small molecule drug, an antibody, an antibody-drug conjugate (ADC), or an aptamer; or a pharmaceutically acceptable salt thereof.

[0204] The AMYR agonist of the invention may be a small molecule drug. As used herein a “small molecule drug” refers to a low molecular weight compound, typically an organic compound. Typically, a small molecule has a maximum molecule weight of 900 Da, allowing for rapid diffusion across cell membranes. The maximum molecular weight of a small molecule may be 500 Da. Typically a small molecule has a size in the order of 1 nm.

[0205] The AMYR agonist of the invention may be an aptamer. Aptamers are generally nucleic acid molecules that bind a specific target molecule. Aptamers can be engineered completely in vitro, are readily produced by chemical synthesis, possess desirable storage properties, and elicit little or no immunogenicity in therapeutic applications. These characteristics make them particularly useful in pharmaceutical and therapeutic utilities.

[0206] As used herein, "aptamer" refers in general to a single or double stranded oligonucleotide or a mixture of such oligonucleotides, wherein the oligonucleotide or mixture is capable of binding specifically to a target. Oligonucleotide aptamers will be discussed here, but the skilled reader will appreciate that other aptamers having equivalent binding characteristics can also be used, such as peptide aptamers. In general, aptamers may comprise oligonucleotides that are at least 5, at least 10 or at least 15 nucleotides in length. Aptamers may comprise sequences that are up to 40, up to 60 or up to 100 or more nucleotides in length. For example, aptamers may be from 5 to 100 nucleotides, from 10 to 40 nucleotides, or from 15 to 40 nucleotides in length. Where possible, aptamers of shorter length are preferred as these will often lead to less interference by other molecules or materials. The AMYR agonist of the invention may be an antibody. As used herein, the term antibody encompasses the use of a monoclonal antibody or polyclonal antibody, as well as the antigen-binding fragments of a monoclonal or polyclonal antibody, or a peptide which binds to REV-ERB with specificity. The antibody may be a Fab, F(ab’)2, Fv, scFv, Fd, Fc or dAb.

[0207] The AMYR agonist of the invention may be an antibody-drug conjugate (ADC), in which a pharmaceutically active moiety is conjugated, directly or indirectly (e.g. via a linker) to an antibody or fragment thereof. For example, an AMYR agonist may be an amylin analogue conjugated, directly or indirectly, to an Fc fragment. The linker may be cleavable or non- cleavable.

[0208] Preferably, an AMYR agonist of the invention is a polypeptide. An AMYR agonist polypeptide may be of between about 15 to about 40 amino acids in length, preferably between about 30 to about 40 amino acids in length, most preferably about 37 amino acids in length. For the avoidance of doubt, any and all disclosure herein in relation to AMYR agonists of the invention applies particularly to AMYR agonist polypeptides, and pharmaceutically acceptable salts thereof, unless expressly stated to the contrary.

[0209] AMYR agonist polypeptides

[0210] Throughout this specification, amino acid positions of an AMYR agonist polypeptide (e.g. lipidated AMYR agonist polypeptides) are numbered according to the corresponding position in pramlintide having the sequence set forth in SEQ ID NO: 5.

[0211] Throughout this specification, amino acids are referred to by their conventional three-letter or single-letter abbreviations (e.g. Ala or A for alanine, Arg or R for arginine, etc.). In the case of certain less common or non-naturally occurring amino acids (i.e. amino acids other than the 20 encoded by the standard mammalian genetic code), unless they are referred to by their full name, frequently employed three- or four-character codes are employed for residues thereof, including aMeSer (fS / 2-amino-3-hydroxy-2-methylphenylpropanoic acid), aMePhe (fS 2-amino-2-methyl-3-phenylpropanoic acid), Aib (2-amino-2-methylpropanoic acid), Dab (2,4-diaminobutanoic acid) and y-Glu (y-glutamic acid, yE).

[0212] An AMYR agonist polypeptide of the invention may comprise or consist of an amino acid sequence having at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or more, up to 100% identity to pramlintide (KCNTATCATQRLANFLVHSSNNFGPILPPTNVGSNTY-amide, SEQ ID NO: 5). Preferably, an AMYR agonist polypeptide of the invention may comprise or consist of an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or more, up to 100% identity to pramlintide (KCNTATCATQRLANFLVHSSNNFGPILPPTNVGSNTY-amide, SEQ ID NO: 5).

[0213] AMYR agonist polypeptides disclosed here can be formulated in or chemically conjugated to e.g. a protein, polymeric drug carrier or advance drug delivery system that further enhance the chemical stability and or physical stability and or the circulatory exposure of the AMYR agonist polypeptide.

[0214] In embodiments of any aspect of the invention, the polypeptides (e.g. lipidated polypeptides) of the invention are isolated polypeptides (e.g. isolated lipidated polypeptides).

[0215] Modified amino acids

[0216] An AMYR agonist polypeptide (e.g. lipidated polypeptides) of the invention may comprise one or more amino acid modifications or substitutions compared to the pramlintide sequence (SEQ ID NO: 5). The AMYR agonist polypeptide may comprise one or more non- proteinogenic amino acids, particularly 2,4-diaminobutanoic acid (Dab). Alternatively or in addition, the AMYR agonist polypeptide comprises one or more alpha methyl amino acids, particularly selected from 2-amino-2-methylpropanoic acid (Aib) and / or alpha methyl phenylalanine (aMePhe or aMeF). The reference to aMePhe and aMeF herein refers to ( S)- 2-amino-2-methyl-3-phenylpropanoic acid.

[0217] Lipidation and Linkers

[0218] An AMYR agonist polypeptide of the invention may be lipidated. Preferably, lipidated polypeptides may have extended half-life compared to pramlintide but without the fibrilforming tendency of lipidated pramlintide analogues described in the art. Without being bound by theory, it is thought that the lipid acts as an albumin binding moiety and protects the AMYR agonist polypeptide against clearance and degradation, thereby extending the half-life of the AMYR agonist polypeptide.

[0219] Accordingly, the lipid may comprise a hydrocarbon chain having from 10 to 26 C atoms, e.g. from 14 to 24 C atoms, e.g. from 16 to 22 C atoms. For example, the hydrocarbon chain may contain 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23 or 24 C atoms. In preferred embodiments, the lipid has 18 to 20 C atoms. In particular, the lipid may have 18 C atoms or 20 C atoms. The hydrocarbon chain may be linear or branched, and may be saturated or unsaturated. Furthermore, it can include a functional group at the end of the lipophilic chain, e.g. a carboxylic acid group which may or may not be protected during synthesis.

[0220] Preferably, the AMYR agonist polypeptide of the invention may be lipidated with a lipid is selected from the group consisting of C12diacid, C14diacid, C16diacid, C17diacid, C18diacid, C19diacid or C20diacid. In some particularly preferred embodiments, the lipid is C18diacid or C20diacid.

[0221] The AMYR agonist polypeptide may comprise at least one lipidated amino acid residue. In some embodiments, the AMYR agonist polypeptide comprises at least two lipidated amino acid residues. Preferably, the AMYR agonist polypeptide may contain only one lipidated amino acid residue. The lipid may be attached to an amino acid residue of the polypeptide.

[0222] The lipid may be attached to the amino acid residue through a linker (referred to herein as "linker-lipid"). Alternatively, the lipid may be directly attached to the amino acid residue without an intervening linker. The lipid may be attached to the amino acid residue via an ester, a sulfonyl ester, a thioester, an amide, an amine or a sulphonamide. Accordingly, it will be understood that the lipid or the linker includes an acyl group, a sulphonyl group, an N atom, an O atom or an S atom which forms part of the ester, sulphonyl ester, thioester, amide, amine or sulphonamide. Optionally, an acyl group in the lipid or linker forms part of an amide or ester with the amino acid residue. Accordingly, preferably the lipid may be attached to an acylation site on the amino acid residue.

[0223] The linker may be attached to any residue of the AMYR agonist polypeptide. The linker may be attached to the side chain of an amino acid residue in the polypeptide, for example to the E-N of a lysine residue. Preferably, the linker is attached to the N-terminus of the polypeptide, (e.g. to a lysine at the N-terminus of the polypeptide). The linker may comprise one or more residues of any naturally occurring or non-naturally occurring amino acid. The linker may comprise a combination of residues, as single or repeating units. For example, the linker may comprise multiple combinations of residues, as single or repeating units, each of which may independently be a residue of Glu (E), y-Glu (yE), Lys, e-Lys, Asp, p-Asp, Gaba, p-Ala (3-aminopropanoyl), O2Oc (2-(2-(2-aminoethoxy)ethoxy)acetic acid), PEG2 (3- (2-(2-aminoethoxy)ethoxy)propanoic acid), PEG4 ( 1 -amino-3, 6, 9, 12-tetraoxapentadecan- 15-oic acid), PEGS (1-amino-3,6,9, 12, 15, 18,21 ,24-octaoxaheptacosan-27-oic acid, PEG12 (1 -amino-3,6,9, 12, 15, 18,21 ,24,27,30,33,36-dodecaoxanonatriacontan-39-oic acid). y-Glu and p-Asp refer to amino acids where the alpha-amino group and the side chain carboxyl group participate in peptide bond formation. c-Lys refers to an amino acid where the epsilon- amino and carboxyl group of lysine participate in peptide bond formation.

[0224] An AMYR agonist polypeptide may comprise a lipid is attached to an amino acid residue in the AMYR agonist polypeptide by a linker, wherein the linker comprises y-glutamic acid (yE), yE-yE, ((O2Oc)-(O2Oc)-yE), ((O2Oc)-(O2Oc)-yEyE), or ((PEG2)-(PEG2)-yE). An AMYR agonist polypeptide may comprise a lipid is attached to an amino acid residue in the AMYR agonist polypeptide by a linker, wherein the linker consists of yE, yE-yE, ((O2Oc)- (020c)- yE), ((O2Oc)-(O2Oc)-yEyE), or ((PEG2)-(PEG2)-yE).

[0225] Combinations of linker, lipid and polypeptide acylation sites are described in WO 2022 / 129254.

[0226] In some preferred embodiments, the lysine at position 1 of the AMYR agonist polypeptide, or pharmaceutically acceptable salt thereof, is lipidated. In some particularly preferred embodiments, wherein the lysine at position 1 of the AMYR agonist polypeptide is lipidated, the lipid is linked to the lysine via a yE-yE linker, and the lipid is octadecanedioic acid (C18diacid).

[0227] Exemplary AMYR agonist polypeptides

[0228] In some preferred embodiments, the lipidated polypeptides of WO 2022 / 129254 may be used in the therapies of the present invention. Thus, an AMYR agonist polypeptide, or pharmaceutically acceptable salt thereof, may comprise or consist of an amino acid sequence having at least 90%, at least 95 at least 97%, at least 98%, at least 99% or more, up to 100% identity to an amino acid sequence selected from the group consisting of:

[0229] C18diacid-yE-[CNTATC]ATQRLAEFLRHSSNN(aMePhe)GPILPPTNVGSNTY-amide (SEQ ID NO: 6);

[0230] K(yE-yE-C18diacid)[CNTATC]ATQRLAEFLRHSSNN(aMePhe)GPILPPTNVGSNTY-amide (SEQ ID NO: 7);

[0231] K(yE-C18diacid)K[CNTATC]ATQRLAEFLRHSSNN(aMePhe)GPILPPTNVGSNTY-amide (SEQ ID NO: 8);

[0232] K(yE-C18diacid)K[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 9);

[0233] K(O2Oc-O2Oc-yE-18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY- amide (SEQ ID NO: 10); K(YE-YE-C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 11 );

[0234] K(YE-C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTNVGSNTY-amide (SEQ ID NO: 12);

[0235] K(YE-YE-C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTNVGSNTY-amide (SEQ ID NO: 13);

[0236] K(YE-YE-C18diacid)[CNTATC]ATQRLANFLVHSSNN(aMePhe)GPILPPTNVGSRTY-amide (SEQ ID NO: 14);

[0237] K(YE-YE-C18diacid)[CNTATC]ATQRLANFLVHSSNN(aMePhe)GPILPPTRVGSNTY-amide (SEQ ID NO: 15);

[0238] K(YE-C18diacid)[CNTATC]ATQRLANFLVHSSNN(aMePhe)GPILPPTRVGSNTY-amide (SEQ ID NO: 16);

[0239] K(YE-C18diacid)K[CNTATC]ATQRLA(Dab)FLVHSSNNFGPILPPTNVGSNTY-amide (SEQ ID NO: 17);

[0240] K(YE-C18diacid)K[CNTATC]ATQRLANFLVHSS(Aib)NFGPILPPTHVGSNTY-amide (SEQ ID NO: 18);

[0241] K(YE-C18diacid)[CNTATC]ATQRLANFLRHSS(Aib)NFGPILPPTEVGSNTY-amide (SEQ ID NO: 19);

[0242] K(YE-C18diacid)[CNTATC]ATQRLANFLRHSS(Aib)NFGPILPPTNVGSNTY-amide (SEQ ID NO: 20);

[0243] K(YE-YE-C18diacid)K[CNTATC]ATQRLANFLRHSS(Aib)NFGPILPPTNVGSNTY-amide (SEQ ID NO: 21 );

[0244] K(YE-YE-C18diacid)K[CNTATC]ATQRLANFLVHSS(Aib)NFGPILPPTRVGSNTY-amide (SEQ ID NO: 22);

[0245] K(YE-C18diacid)K[CNTATC]ATQRLANFLVHSS(Aib)NFGPILPPT(Aib)VGSNTY-amide (SEQ ID NO: 23);

[0246] K(YE-C18diacid)K[CNTATC]ATQRLANFL(Aib)HSSNNFGPILPPTNVGSNTY-amide (SEQ ID NO: 24); and K(YE-C18diacid)K[CNTATC]ATQRLA(Dab)FL(Aib)HSSNNFGPILPPTEVGSNTY-amide (SEQ ID NO: 25).

[0247] In some preferred embodiments, an AMYR agonist polypeptide comprises an amino acid sequence having at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or more, up to 100% identity to the amino acid sequence K[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 26). In some preferred embodiments, an AMYR agonist polypeptide consists of an amino acid sequence having at least 70%, at least 80%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99% or more, up to 100% identity to the amino acid sequence K[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 26).

[0248] In some particularly preferred embodiments, an AMYR agonist polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99% or more, up to 100% identity to the amino acid sequence K[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 26).

[0249] In some particularly preferred embodiments, an AMYR agonist polypeptide consists of an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99% or more, up to 100% identity to the amino acid sequence K[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 26).

[0250] More preferably, the AMYR agonist polypeptide, or pharmaceutically acceptable salt thereof, comprises the amino acid sequence K(yE-yE-

[0251] C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 11).

[0252] Most preferably, the AMYR agonist polypeptide, or pharmaceutically acceptable salt thereof, consists of the amino acid sequence K(yE-yE- C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 11).

[0253] In any one of the methods disclosed herein, the AMYR agonist may be a polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence K(yE-yE- C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 11) in an amount of about 1.5 mg. The subject may have obesity or an obesity-related condition.

[0254] In any one of the methods disclosed herein, the AMYR agonist may be a polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence K(yE-yE- C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 11) in an amount of about 4.5 mg. The subject may have obesity or an obesity-related condition. In any one of the methods disclosed herein, the AMYR agonist may be a polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence K(yE-yE- C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 11 ) in an amount of about 9 mg. The subject may have obesity or an obesity-related condition.

[0255] In any one of the methods disclosed herein, the AMYR agonist may be a polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence K(yE-yE- C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 11 ) in an amount of about 1 .5 mg. The subject may have a metabolic disease.

[0256] In any one of the methods disclosed herein, the AMYR agonist may be a polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence K(yE-yE- C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 11) in an amount of about 4.5 mg. The subject may have a metabolic disease.

[0257] In any one of the methods disclosed herein, the AMYR agonist may be a polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence K(yE-yE- C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 11) in an amount of about 9 mg. The subject may have a metabolic disease.

[0258] Pharmacokinetics

[0259] The AMYR agonist polypeptides used in the invention may exhibit favourable pharmacokinetic properties as compared to pramlintide. For example, the AMYR agonist polypeptides may have an extended half-life as compared to pramlintide.

[0260] As used herein, the term "half-life" is used to refer to the time taken for the concentration of isolated polypeptide in plasma to decline to 50% of its original level. Methods to determine the half-life of proteins are known in the art and are described in Example 4 of WO 2022 / 129254. It will be recognised that an extended half-life is advantageous, as it permits the therapeutic proteins to be administered according to a safe and convenient dosing schedule, e.g. lower doses that can be administered less frequently. Moreover, the achievement of lower doses may provide further advantages such as the provision of an improved safety profile. To the contrary, pramlintide requires frequent and inconvenient administration. The AMYR agonist polypeptides may have a half-life of at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10 hours, at least 11 hours, at least 12 hours, at least 13 hours or at least 14 hours in rat models. Preferably, the AMYR agonist polypeptides may have a half-life of at least 14 hours.

[0261] Reduced Fibrilation

[0262] The AMYR agonist polypeptides may exhibit reduced tendency to undergo fibrillation in pharmaceutically relevant aqueous media, especially at pH values in the range from 4 to 7, as compared to lipidated pramlintide. The AMYR agonist polypeptides may exhibit reduced tendency to undergo fibrillation in pharmaceutically relevant aqueous media, especially at pH values in the range from 4 to 7, as compared to pramlintide which is lipidated in a similar manner e.g. the same lipid is attached, the lipid is attached through the same linker and / or the lipid is attached at the same position. Exemplary lipidated AMYR agonist polypeptides, include, for example, SEQ ID NOs: 6-25.

[0263] Accordingly, the AMYR agonist polypeptides may be suited for formulation in acidic media (e.g. pH 4) and in neutral or near-neutral media (e.g. pH 7 or 7.4). Such AMYR agonist polypeptides may be well suited for coformulation with, for example, insulin, various insulin analogues and / or other therapeutic (e.g. anti-diabetic or anti-obesity) agents that require a neutral or near-neutral formulation pH.

[0264] The AMYR agonist polypeptides may show no detectable fibrillation after about 5 hours, after about 7 hours, after about 9 hours, after about 11 hours, after about 13 hours, after about 15 hours, after about 17 hours or after about 20 hours, after about 48 hours, after about 72 hours, after about 96 hours, after about 108 hours, after about 120 hours, after 132 about hours or after about 144 hours at pH 4 and 37°C. Suitable assays for quantifying fibrillation are described in Example 3 of WO 2022 / 129254 (which describes a Thioflavin T fibrillation assay).

[0265] Preferably, the AMYR agonist polypeptides are soluble at concentrations required for therapeutic efficacy.

[0266] Pharmaceutical compositions An AMYR agonist of the invention may be provided in a pharmaceutical composition. As described herein, said AMYR agonist is typically an agonist polypeptide.

[0267] Further provided are compositions, e.g., pharmaceutical compositions, that contain an effective amount of an AMYR agonist of the invention as provided herein e.g. about 1 mg to about 15 mg of AMYR agonist). The compositions may be formulated for the treatment of obesity, an obesity-related condition or a metabolic disease as described herein, e.g., obesity and / or type 2 diabetes.

[0268] The pharmaceutical compositions of the invention may comprise one or more excipient(s), carrier, buffer, stabilizer or other materials well known to those skilled in the art. Such materials should be non-toxic and should not interfere with the efficacy of the active ingredient. The precise nature of the carrier or other material will depend on the route of administration. Pharmaceutically acceptable excipients are known in the art, see for instance Remington's Pharmaceutical Sciences (by Joseph P. Remington, 18th ed., Mack Publishing Co., Easton, PA), which is incorporated herein in its entirety.

[0269] Composition can be in a variety of forms, including, but not limited to an aqueous solution, an emulsion, a gel, a suspension, lyophilized form, or any other form known in the art. In addition, the composition can contain pharmaceutically acceptable additives including, for example, diluents, binders, stabilizers, and preservatives. Once formulated, compositions of the invention can be administered directly to the subject.

[0270] For injectable formulations, e.g., for subcutaneous injection, the active ingredient will be in the form of a parenterally acceptable aqueous solution which is pyrogen-free and has suitable pH, isotonicity and stability.

[0271] An AMYR agonist as described herein may be formulated in liquid, semi-solid or solid forms depending on the physicochemical properties of the molecule and the route of delivery. Formulations may include excipients, or combinations of excipients, for example: sugars, amino acids and surfactants. Liquid formulations may include a wide range of antibody concentrations and pH. Solid formulations may be produced by lyophilisation, spray drying, or drying by supercritical fluid technology, for example. Treatment may be given by injection (for example, subcutaneously, or intra-venously. The treatment may be administered orally. The route of administration can be determined by the physicochemical characteristics of the treatment, by special considerations for the disease or by the requirement to optimize efficacy or to minimize side-effects. A preferred route of administering pharmaceutical compositions of the present invention is subcutaneous administration.

[0272] The pharmaceutical composition may be a solid composition. Such solid compositions may be reconstituted prior to administration. The pharmaceutical composition may be a liquid composition.

[0273] A composition may be administered alone or in combination with other treatments, either simultaneously or sequentially dependent upon the condition to be treated.

[0274] An AMYR agonist may be used as part of a combination therapy in conjunction with an additional medicinal component. Combination treatments may be used to provide significant synergistic effects, particularly the combination of an AMYR agonist with one or more other drugs. For example, an AMYR agonist may be administered with one or more other antiobesity drugs, such as an incretin mimetic drug e.g., a GLP-1 R agonist) and / or a leptin. An AMYR agonist may be administered concurrently or sequentially or as a combined preparation with another therapeutic agent or agents, for the treatment of one or more of the conditions listed herein.

[0275] Methods of production

[0276] The polypeptides of the invention, particularly the AMYR agonist polypeptides, may be produced by any method known in the art.

[0277] The production of polypeptides such as AMYR agonist polypeptides is well known in the art. The AMYR agonist polypeptides of the invention can thus be produced by chemical synthesis, e.g. solid phase polypeptide synthesis using t-Boc or Fmoc chemistry, or other well-established techniques as described by Merrifield (1963, J. Am. Chem. Soc. 85:2149- 2154). Solid phase peptide synthesis can be accomplished, e.g., by using automated synthesizers, using standard reagents, e.g., as explained in Example 1 of WO 2014 / 091316, which is herein incorporated by reference in its entirety.

[0278] Alternatively, AMYR agonist polypeptides provided herein can be produced recombinantly using a convenient vector / host cell combination as would be well known to the person of ordinary skill in the art, e.g., by recombinant expression of a nucleic acid molecule encoding a fusion polypeptide in a host cell. Generally, a polynucleotide sequence encoding the polypeptide is inserted into an appropriate expression vehicle, e.g., a vector which contains the necessary elements for the transcription and translation of the inserted coding sequence. The nucleic acid encoding the polypeptide is inserted into the vector in proper reading frame. The expression vector is then transfected into a suitable host cell which will express the polypeptide. Suitable host cells include without limitation bacteria, yeast, or mammalian cells. A variety of commercially-available host-expression vector systems can be utilized to express the polypeptides described herein.

[0279] Following synthesis, the AMYR agonist polypeptides may optionally be isolated or purified.

[0280] Kits and Articles of Manufacture

[0281] The present invention further provides a kit comprising AMYR agonist polypeptides or pharmaceutical compositions of the invention. The AMYR agonist polypeptide is typically provided in one or more dose of about 1 mg to about 15 mg. The kit may comprise a package containing the AMYR agonist polypeptides or pharmaceutical composition thereof with instructions. The instructions may be directed to use of the AMYR agonist polypeptide for reducing weight in a subject at a dose of about 1 mg to about 15 mg. The instructions may be directed to use of the AMYR agonist polypeptide for treating or preventing obesity or an obesity-related condition at a dose of about 1 mg to about 15 mg. The instructions may be directed to use of the AMYR agonist polypeptide for treating or preventing a metabolic disease at a dose of about 1 mg to about 15 mg.

[0282] In said kit, the AMYR agonist polypeptide or pharmaceutical composition of the invention may be formulated in single dose vials or a container closure system (e.g. pre- filled syringe). Optionally associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration.

[0283] The present invention also provides an article of manufacture comprising AMYR agonist polypeptides or pharmaceutical compositions of the invention. The AMYR agonist polypeptide is typically provided in one or more dose of about 1 mg to about 15 mg.

[0284] Brief Description of Sequence Listing Where present, a square bracket [ ] between the two cysteine residues (Cys2 and Cys7) indicate the presence of an intramolecular disulphide bridge. yE or yE = gamma-glutamate

[0285] SEQ ID NO: 1 exemplary amino acid sequence for hRAMPI

[0286] SEQ ID NO: 2 exemplary amino acid sequence for hRAMP2

[0287] SEQ ID NO: 3 exemplary amino acid sequence for hRAMP3

[0288] SEQ ID NO: 4 exemplary amino acid sequence for hCTR

[0289] SEQ ID NO: 5 pramlintide amino acid sequence

[0290] SEQ ID NO: 6 C18diacid-yE-

[0291] [CNTATC]ATQRLAEFLRHSSNN(aMePhe)GPILPPTNVGSNTY-amide

[0292] SEQ ID NO: 7 K(yE-yE-

[0293] C18diacid)[CNTATC]ATQRLAEFLRHSSNN(aMePhe)GPILPPTNVGSNTY-amide

[0294] SEQ ID NO: 8 K(yE-

[0295] C18diacid)K[CNTATC]ATQRLAEFLRHSSNN(aMePhe)GPILPPTNVGSNTY-amide);

[0296] SEQ ID NO: 9 K(yE-

[0297] C18diacid)K[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide

[0298] SEQ ID NO: 10 K(O2Oc-O2Oc-yE-

[0299] 18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide

[0300] SEQ ID NO: 11 K(yE-yE-

[0301] C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide

[0302] SEQ ID NO: 12 K(yE-

[0303] C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTNVGSNTY-amide

[0304] SEQ ID NO: 13 K(yE-yE-

[0305] C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTNVGSNTY-amide

[0306] SEQ ID NO: 14 K(yE-yE-

[0307] C18diacid)[CNTATC]ATQRLANFLVHSSNN(aMePhe)GPILPPTNVGSRTY-amide

[0308] SEQ ID NO: 15 K(yE-yE-

[0309] C18diacid)[CNTATC]ATQRLANFLVHSSNN(aMePhe)GPILPPTRVGSNTY-amide

[0310] SEQ ID NO: 16 K(yE-

[0311] C18diacid)[CNTATC]ATQRLANFLVHSSNN(aMePhe)GPILPPTRVGSNTY-amide

[0312] SEQ ID NO: 17 K(yE-

[0313] C18diacid)K[CNTATC]ATQRLA(Dab)FLVHSSNNFGPILPPTNVGSNTY-amide

[0314] SEQ ID NO: 18 K(yE-

[0315] C18diacid)K[CNTATC]ATQRLANFLVHSS(Aib)NFGPILPPTHVGSNTY-amide SEQ ID NO: 19 K(yE-

[0316] C18diacid)[CNTATC]ATQRLANFLRHSS(Aib)NFGPILPPTEVGSNTY-amide

[0317] SEQ ID NO: 20 K(yE-

[0318] C18diacid)[CNTATC]ATQRLANFLRHSS(Aib)NFGPILPPTNVGSNTY-amide

[0319] SEQ ID NO: 21 K(YE-YE-

[0320] C18diacid)K[CNTATC]ATQRLANFLRHSS(Aib)NFGPILPPTNVGSNTY-amide

[0321] SEQ ID NO: 22 K(YE-YE-

[0322] C18diacid)K[CNTATC]ATQRLANFLVHSS(Aib)NFGPILPPTRVGSNTY-amide

[0323] SEQ ID NO: 23 K(YE-

[0324] C18diacid)K[CNTATC]ATQRLANFLVHSS(Aib)NFGPILPPT(Aib)VGSNTY-amide

[0325] SEQ ID NO: 24 K(YE-

[0326] C18diacid)K[CNTATC]ATQRLANFL(Aib)HSSNNFGPILPPTNVGSNTY-amide

[0327] SEQ ID NO: 25 K(YE-

[0328] C18diacid)K[CNTATC]ATQRLA(Dab)FL(Aib)HSSNNFGPILPPTEVGSNTY-amide

[0329] SEQ ID NO: 26 K[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY- amide

[0330] EXAMPLES

[0331] The invention is now described with reference to the Examples below. These are not limiting in terms of the scope of the invention, and a person skilled in the art would appreciate that suitable equivalents could be used within the scope of the present invention. Thus, the Examples may be considered component parts of the invention, and the individual aspects described therein may be considered as disclosed independently, or in any combination.

[0332] Example 1 : A Phase I Single Ascending Dose (SAD) study and a Phase lb repeated dose study of administering an AMYR agonist to healthy subjects who are overweight or obese

[0333] Pre-clinical studies have demonstrated that an AMYR agonist promotes, maintains and enhances weight loss without compromising tolerability in animal models. In particular, 20 nmol / kg of an AMYR agonist results in a reduction in overall body weight loss (in grams) in diet-induced obese (DIO) rats in a 15 day treatment as compared to vehicle (data not shown). In particular, the AMYR agonist was found to drive good quality weight loss by inducing more fat mass specific body weight loss and preserving more lean muscle mass as compared to a GLP-1 receptor agonist drug, semaglutide (10nmol / kg), and rats pair-fed to the AMYR agonist (i.e., rats having restricted food intake to match the treatment group diet). In a separate experiment, a AMYR agonist (30 nmol / kg) was found to be highly tolerable as measured by rodent aversion studies (an experiment in which a rodent’s aversion to saccharin following administration of a drug is a correlate for nausea), in which the AMYR agonist demonstrated significantly less aversion in comparison to vehicle as compared to a DACRA (cagrilintide 10 nmol / kg, p<0.05), a GLP-1 receptor agonist (semaglutide 2.4 nmol / kg, p<0.01), and a combination thereof (p<0.0001 ).

[0334] Following the pre-clinical studies, a Phase I randomised, single-blind, placebo-controlled study was performed in order to assess the safety, tolerability, and pharmacokinetics of an AMYR agonist following single ascending dose (SAD) administration to healthy subjects who are overweight or obese (trial identifier: D8750C00001 ; ClinicalTrials.gov identifier: NCT05511025). The inclusion criteria for the study was healthy male and female participants aged 18 to 55 years and a body mass index (BMI) of 25 to 35 kg / m2inclusive (at the time of screening) and weighing at least 50 kg. A Japanese cohort was also designed to include natives of Japan (defined as having both parents and four grandparents who are Japanese, including second and third generation subjects of Japanese descent whose parents or grandparents are living in a country other than Japan) and having a BMI of 23 to 35 kg / m2inclusive (at the time of screening) and weighing at least 50 kg.

[0335] In the study, 54 participants were dosed with a single dose of between 0.3 mg to 4.2 mg of an AMYR agonist or placebo by subcutaneous administration, a single dose of 0.3 mg by intravenous administration, or a single dose of 2.7 mg by subcutaneous administration in the Japanese cohort. Subjects were monitored for a period of 15 days. The AMYR agonist was found to demonstrate good efficacy with competitive tolerability. In particular, the AMYR agonist demonstrated dose dependent weight loss (approximately 1 .5% - 3% mean body weight change from baseline) at doses of 0.3 mg, 0.9 mg, 1 .5 mg, 2.7 mg, and 4.2 mg. Peak weight loss was demonstrated at 4-5 days in all groups. Strong weight loss profiles were seen in the 1.5 mg and 4.2 mg cohorts in particular. Following a single 2.7 mg dose of AMYR agonist, a mean weight loss of 1 .7% was seen in the global cohort (s.c.), and 3.8% in the Japanese cohort (s.c.; see Figure 1).

[0336] The AMYR agonist also demonstrated a good tolerability profile, in which no safety concerns were identified at single doses up to 4.2 mg. Moderate or severe vomiting were only demonstrated for some subjects in the 4.2 mg group (s.c.), while some incidences of mild nausea and / or mild vomiting occurred in the 1.5 mg and 2.7 mg (global and Japanese cohorts) groups (see Figure 2). A Phase lb study designed to assess the safety, tolerability, pharmacokinetics, and the impact on body weight of repeated doses of an AMYR agonist in healthy participants with overweight or obesity (trial identifier: D8750C00002; ClinicalTrials.gov identifier: NCT06132841 ) is currently ongoing. The inclusion criteria for the study is healthy male and female participants aged 18 to 55 years and a body mass index (BMI) of 25 to 40 kg / m2inclusive (at the time of screening) and weighing at least 60 kg.

[0337] The study involves a treatment period of 6 weeks (cohort 1 ) and 12 weeks (cohort 2 and 3) during which the participants receive once-weekly repeated doses of the drug at doses between 1.5 mg to 9 mg during residency at a clinical unit. Cohort 1 (n=15+5) receive repeated doses of 2.7 mg of the drug, or placebo, over six weeks. Cohorts 2 (n=15+5) and 3 (n=9+3) receive repeated doses gradually titrated from an initial 1 .5 mg to 9 mg by the end of the study over a 12 week period, or placebo. Eligible participants were randomized to AMYR agonist and placebo in a 3:1 ratio.

[0338] Based on interim data from a subset of the participants following s.c. weekly administration of between 1.5 mg to 9 mg AMYR agonist, the AMYR agonist was well tolerated and no safety concerns were identified. In particular, following 6 weeks of treatment with 2.7 mg (Cohort 1 ), 2.1% weight loss (3.5 % placebo-corrected) was measured following 6 weeks of weekly repeat dosing (see Figure 3A). Further, four (27%) and one (6%) participants on active treatment of 2.7 mg reported mild to moderate nausea and vomiting, respectively. In particular, the sole report of vomiting occurred within a week of the first dose, all reports of moderate nausea occurred within a week of the first dose, with only one report of mild nausea beyond the second dose. Thus, the vast majority of these incidents occurred in the first week after the first dose (see Figure 4A).

[0339] Following 12 weeks of treatment gradually titrated from 1.5 mg to 9 mg (Cohort 2), 4% weight loss (3.8% placebo-corrected) was measured (see Figure 3B). Three participants reported mild to moderate nausea (20%) and one reported mild vomiting (6%) following 12 weeks of weekly repeat dosing. The sole report of vomiting occurred shortly after the escalation to a 9 mg dose (see Figure 4B).

[0340] These studies demonstrate a dose-dependent reduction in bodyweight across the dose range of 1 .5 to 9 mg in humans, which based on preclinical models is expected to be almost exclusively due to fat mass loss. Further, it was observed that even relatively high doses of AMYR agonist therapy are surprisingly well tolerated in human subjects. In these studies, the AMYR agonist was AZD6234 (SEQ ID NO: 11). AZD6234 is a potent agonist of human, cynomolgus monkey, and rat amylin receptors, displaying in vitro selectivity over calcitonin receptor activation.

[0341] Example 2: A Phase lib Randomised, Double-blind, Placebo-controlled Study to Evaluate the Efficacy, Safety and Tolerability of an AMYR agonist in Participants Living with Obesity or Overweight with Comorbidity

[0342] A Phase lib, global, randomised, parallel-group, double-blind, placebo-controlled, multicentre study (‘APRICUS’ study) was designed for human subjects (approximately 231 participants) between 18 to 75 years of age with BMI > 30 kg / m2, or BMI > 27 kg / m2with at least one weight-related comorbidity (hypertension, dyslipidaemia or hyperlipidaemia, CV disease or obstructive sleep apnoea). The study was designed to be conducted at up to 50 sites in up to 6 countries.

[0343] An eligibility screening protocol was designed. The eligibility screening period is up to 28 days prior to randomisation. The screening protocol was designed to include eligible consenting participants according to age (18 to 75 years of age, inclusive, at the time of signing the informed consent, with suitable veins for cannulation or repeated venipuncture), disease characteristics (at screening, have a BMI of (a) > 30 kg / m2inclusive, or (b) > 27 kg / m2and at least one of the following comorbidities (with documented medical history or currently being treated for): (i) Hypertension (ii) Dyslipidaemia or hyperlipidaemia (iii) CV disease and / or (iv) Obstructive sleep apnoea), weight (a stable body weight for 3 months prior to screening (± 5% body weight change)), contraceptive requirements, and a negative pregnancy test (at screening and randomisation) and non-breastfeeding with respect to female participants of childbearing potential. Exclusion criteria included weight criteria, including subjects having obesity induced by other endocrine disorders (such as Cushing’s syndrome or Prader-Willi syndrome), subjects receiving prescription or non-prescription medication for weight loss within the 3 months prior to screening, and / or previous or planned (within study period) bariatric surgery or fitting of a weight loss device (e.g. gastric balloon or duodenal barrier); diabetes status, including a history of type 1 or type 2 diabetes mellitus, treatment with diabetes medication in past 3 months prior to screening, and / or a haemoglobin (HbA1c) level > 6.5% (48 mmol / mol) at screening; gastrointestinal health status (e.g., gastroparesis and / or IBD); cardiovascular / renal health status {e.g., hypertension); mental health status; other disease or diagnostic status e.g., HIV and / or hepatitis B antibody); prior or concomitant therapy associated with weight gain or that deemed clinically relevant; and prior or concomitant clinical study experience.

[0344] Following screening, eligible participants are randomised to receive 1.5 mg (no titration), 4.5 mg (following titration a or b), or 9 mg (following titration a or b) of AMYR agonist or matching placebo (following no titration, or titration a or b). A simplified version of the study design is set out at Figure 5. The titration schedule is set out at Table 1. Participants are randomised to receive one of the 3 AMYR agonist doses or volume-matched placebo, in a 3:2 ratio for the 1.5 mg AMYR agonist dose and in a 3:1 ratio for the 4.5 mg and 9.0 mg AMYR agonist doses. Pooling by dose level, the randomisation for (1.5 mg AMYR agonist):(4.5 mg AMYR agonist):(9.0 mg AMYR agonist):(placebo) is 1 :2:2:2.

[0345] Based on the weight loss, tolerability, safety, and pharmacokinetic data from the Phase I SAD and the Phase lb studies, the Phase lib trial was designed to examine the effect of 3 target doses of AMYR agonist (1 .5 mg, 4.5 mg, and 9 mg) or placebo on body weight over an initial period of 26 weeks and on to the end of treatment at 36 weeks. The primary endpoint was designed to be evaluated after all participants complete 26 weeks. The doses in the trial do not exceed the highest dose achieved in the clinical Phase lb study.

[0346] The top dose in this trial (9 mg) was selected based on combined consideration of weight loss, tolerability, and safety. The 9 mg dose is at the plateau of the predicted exposureresponse for weight loss based on modelling. The 9 mg dose was tolerated and no safety concerns raised in Phase lb following titration. Based on the weight loss data for AZD6234 and modelling, it is expected that doses of 1.5, 4.5, and 9 mg will provide adequate information on weight loss to establish dose- and / or exposure relationship for weight loss to support dose selection for a Phase III trial.

[0347] The starting dose of 1 .5 mg for all treatment arms was selected since it was shown to be well tolerated with only mild Gl-related tolerability issues in the Phase I SAD / Phase lb studies reported in Example 1. To reach higher doses than 1.5 mg, the dose will be gradually increased to reach 4.5 mg and 9 mg (according to Table 1).

[0348] The study is double-blind and each AMYR agonist treatment group re placebo-matched with respect to titration schedule and dose levels.

[0349] Each treatment with the AMYR agonist or placebo is administered by subcutaneous injection. Administration will either be carried out at the clinic, or by self-administration. Determination of subjects permitted to self-administer is determined by the medical team, who provide instructions and training for self-administration, including injection site selection and rotation.

[0350] Participants begin treatment (placebo or AMYR agonist) on Day 1 and continue for 36 weeks. After the treatment period, participants will have one follow-up visit approximately 7 weeks post last dose.

[0351] Body weight was designed to be measured in the morning while the participant is fasted and prior to breakfast. At screening (Visit 1) and all clinic visits, the participant’s weight is measured without shoes, on an empty bladder and only wearing lightweight clothing (no heavy jumpers / sweaters / outer garments); all readings are to be recorded as accurately as possible on a digital scale and recorded in kilograms (one decimal). The same model scale is to be used for body weight measurement throughout the study and calibrated on a regular basis as recommended by the manufacturer.

[0352] Synthesis of AMYR agonists, such as lipidated pramlintide analogue polypeptides, have been described elsewhere (see WO 2022 / 129254, the contents of which are hereby incorporated by reference). In this clinical trial, the AMYR agonist is AZD6234 (SEQ ID NO: 11 ). AZD6234 is a potent agonist of human, cynomolgus monkey, and rat amylin receptors, displaying in vitro selectivity over calcitonin receptor activation. In vitro studies and in vivo toxicology studies demonstrated acceptable safety profiles.

[0353] Table 1 Titration Regimen

[0354] 1, low dose / no titration; 2a, medium dose; 2b, medium dose; 3a, high dose; 3b, high dose. Primary objectives

[0355] The co-primary objectives of this study are:

[0356] • Percent change in body weight from baseline to Week 26

[0357] • Proportion of participants with weight loss > 5% from baseline at Week 26

[0358] The secondary objectives of this study are:

[0359] • Proportion of participants with weight loss > 5% from baseline at Week 36

[0360] • Proportion of participants with weight loss > 10% from baseline at Week 26

[0361] • Proportion of participants with weight loss > 10% from baseline at Week 36

[0362] • Percent change in body weight from baseline to Week 36

[0363] • Absolute change in body weight (kg) from baseline to Week 26 and Week 36

[0364] • AZD6234 plasma concentration (characterising drug pharmacokinetics)

[0365] The safety and tolerability of AZD6234 is to be assessed by monitoring incidence of adverse events, vital signs, clinical and laboratory assessments, and ECG. Adverse effects (AEs) include serious AEs (SAEs), SAEs with outcome of death, AEs leading to discontinuation of AZD6234, AEs leading to dose reduction of AZD6234, and AEs possibly related to AZD6234. In particular, AEs include nausea and vomiting, which will also be summarised by titration arm. Vital signs include systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR). Laboratory parameters include Clinical Chemistry and Haematology parameters as well as urinalysis. ECG measurements include HR, Time Duration Between Consecutive R-waves (RR), Pulmonic (Pulmonary) Regurgitation (PR), QRS Complex (Q-, R-, and S-wave; QRS), and QT intervals. Derived variables include Absolute QT Interval Corrected for Heart Rate by Fridericia's formula (QTcF). The immunogenicity of AZD6234 is to be measured by ADA prevalence, incidence and titre.

[0366] Body composition as measured by bioimpedance and waist circumference (WC):

[0367] • Change and percentage change in BMI, total body mass, body fat, muscle mass, and fat free mass in kg and change in WC in cm from baseline to Week 26 and Week 36

[0368] Glucose homeostasis, / 3-cell function, and insulin sensitivity:

[0369] • Change from baseline in HOMA2 scores (HOMA2-IR, HOMA2-%B, and HOMA2-%S) at Week 26 and Week 36

[0370] • Change from baseline in HbA1c, fasting serum glucose, fasting serum insulin, fasting serum c-peptide, and plasma proinsulin levels at Week 26 and Week 36

[0371] End-organ protection:

[0372] • Change from baseline to Week 26 and Week 36 on CV endpoints:

[0373] • ApoB / ApoA1 , hs Troponin T)

[0374] • Change from baseline to Week 26 and Week 36 on renal endpoints:

[0375] • eGFR (creatinine and cystatin C)

[0376] • UACR (urine albumin and creatinine ratio)

[0377] • Renin (part of safety assessment)

[0378] • Change from baseline to Week 26 and Week 36 on liver endpoints:

[0379] • ALT, AST, FIB-4 (calculated AST, ALT, age, platelet)

[0380] Weight-loss quality:

[0381] • Change from baseline to Week 26 and Week 36 on metabolic / adipokine health:

[0382] • Adiponectin, Leptin

[0383] • Lipid profile (TG, total cholesterol, NEFA, HDL, LDL, (calculated) VLDL (calculated)

[0384] • Change from baseline to Week 26 and Week 36 on systemic inflammation:

[0385] • hsCRP, hslL6

[0386] Nausea:

[0387] • Change from baseline in GDF15 to Week 4, 12, 26, and 36 Treatment response predictions versus placebo

[0388] • Investigate associations between: Demographics, standard clinical chemistry, and other included biomarkers with response (BMI)

[0389] • Explore response in post-menopausal women as a sub-segment

[0390] Exploratory biomarkers of Mechanism of Action:

[0391] • Change from baseline to Week 26 and Week 36 in exploratory MoA by analysing plasma samples using Olink panels.

[0392] Blood pressure and heart rate:

[0393] • Change from baseline to the end of Week 26 in mean SBP and DBP, and HR over 24 hours and during the day and night

[0394] Plasma and serum sample biomarkers:

[0395] • Potential biomarkers in EDTA plasma and serum samples

[0396] Dose / exposure and efficacy / tolerability:

[0397] • Dose / exposure of AZD6234 relative to efficacy (e.g., change form baseline body weight) and tolerability

[0398] Physical function, quality of life and tolerability

[0399] • Change from baseline to Week 26 and Week 36 in IWQoL-Lite CT Physical Function score, SF-36 v2 Physical functioning score, IWQoL-Lite CT Total Score, SF-36 v2 physical component and mental component scores

[0400] • Analysis of FACIT-GP5 scores across assessment timepoints

[0401] Statistical Methods

[0402] Approximately 231 eligible participants are to be randomly assigned the study drug, of which approximately 182 participants are anticipated to complete treatment and assessments through Week 26. This sample size provides > 90% power to determine that each of the 4.5 mg and 9.0 mg AMYR agonist doses (> 80% power for the 1 .5 mg AZD6234 dose) is superior to placebo based on the co-primary endpoint of percentage change from baseline in weight at Week 26, assuming a SD of 6.5%, 2-sided alpha level of 0.05, and a treatment difference of 5%. It also provides > 90% power to determine that each of the 4.5 mg and 9.0 mg AMYR agonist doses (> 80% power for the 1.5 mg AMYR agonist dose) is superior to placebo based on the co-primary endpoint of proportion of participants with weight loss > 5% from baseline at Week 26, assuming 70% response in these AMYR agonist dose arms compared to 30% in the placebo arm and a 2-sided alpha level of 0.05.

[0403] For efficacy analyses, participants on AMYR agonist dose are pooled by dose, regardless of the titration regimen (a or b). Participants on placebo are also pooled. Multiple testing adjustments will not be applied to the comparisons between each dose level and placebo.

[0404] The co-primary endpoint of percent change in body weight from baseline at Week 26, is to be analysed using a mixed model for repeat measures (MMRM) comprising of the fixed effects of baseline weight (kg), target dose, visit, and treatment-by-visit interaction, and the random effect (random intercept) of participant. BMI status, sex, and ethnicity may also be included as fixed effects. The Restricted Maximum Likelihood estimation approach is to be used. The treatment effects will be summarised by the difference in least squares means, 95% confidence intervals (Cis), and p-values.

[0405] The co-primary endpoint of proportion of participants with body weight loss of at least 5% at Week 26, is to be analysed using the Cochran-Mantel-Haenszel test, stratified by sex and baseline BMI status. P-value of the test of general association will be presented. Mantel- Haenszel common risk difference will be obtained along with 95% Cis, based on the Sato variance estimator.

[0406] Example 3: AZD6234 treatment restores leptin-mediated pSTAT3 signaling in the ventromedial hypothalamus of DIO rats when compared to semaglutide

[0407] Sprague Dawley rats arrived at 8 weeks and were sorted into chow diet (lean) and a condensed milk diet (DIO) groups. After 4 weeks on diet, the DIO and lean rats were sorted based on BW into 10 groups with 2 groups each receiving one of the following treatments daily for 2 weeks:

[0408] Lean: Vehicle (Control)

[0409] DIO: Vehicle (Control)

[0410] DIO: AZD6234 (10 nmol / kg)

[0411] DIO: Semaglutide (3 nmol / kg)

[0412] DIO: Pair-fed to AZD6234 Animals received injections QD with 8 rats per group. Doses of the analogs were selected to produce equivalent body weight reductions. On day 10 body composition was measured via NMR. On day 14, after a 4 h fast plasma endogenous leptin was measured (Figure 6A), and each group was given either a PBS or murine leptin (13 mg / kg) challenge. After 45 min, brains were collected and pSTAT3+cells in the ventromedial hypothalamus (VMH) were quantified (Figure 6B).

[0413] The leptin challenge induced pSTAT3 activation in the VMH, with lean rats showing 14.8-fold increase in pSTAT3 positive nuclei vs PBS challenged animals. In DIO rats, vehicle and semaglutide treatment exhibited a blunted response around 4-fold. In contrast, AZD6234 increased pSTAT3+ labeling by 16.8-fold, while pair-fed treated rats showed 11 -fold increase. These results (shown in the Table below and Figure 6C) demonstrate that AZD6234 restores basal levels of pSTAT3 activation in the VMH, while semaglutide does not.

[0414] Post hoc ana ysis conducted using Tukey’s multiple comparison’s test

[0415] Thus, AZD6234 treatment restored leptin-mediated pSTAT3 signaling in the VMH of DIO rats when compared to semaglutide. Further, AZD6234 treatment reduced body fat to the level of aged match lean rats within 14 days, almost doubling the effect seen in animals treated with semaglutide, despite both treatment groups exhibiting similar body weight reductions (data not shown). These data suggest the long-acting amylin analog AZD6234 is a potential option for the treatment of obesity and weight management, and in particular that AZD6234 may be used in combination therapy with leptin for the treatment of obesity and weight management. Example 4: Selective amylin receptor (AMYR) agonist monotherapy reduces body weight and food intake in a dose-dependent manner and drives fat-mass specific body weight loss and preserves lean-mass and reduced aversion as compared to a dual amylin and calcitonin receptor agonist (DACRA) monotherapy in DIO rats

[0416] Study 1 Design: High-fat diet (DIO) fed rats were used to profile AMYR agonist or DACRA monotherapy to evaluate food intake, body weight and body composition effects in a 4-week study. In this experiment, the AMYR agonist was that of SEQ ID NO: 11 and the DACRA was cagrilintide.

[0417] Study 1 Data: AMYR agonist (2.5 or 10 nmol / kg) or DACRA (1 , 3, 10, or 20 nmol / kg) over a 4-week study in DIO rats resulted in dose-dependent reduction in body weight compared to vehicle treated rats. BW reduction was -5.2 ± 0.6 and -9.7 ± 0.7 % for AMYR agonist (2.5 or 10 nmol / kg), and -3.0 ± 0.6, -5.3 ± 1.1 , -8.8 ± 1 .4 and -11 .6 ± 0.9 % for DACRA (1 , 3, 10, or 20 nmol / kg), respectively (see Figure 9A). A reduction of cumulative food intake of -14.3 ± 1.2 and -20.9 ± 0.7 % of vehicle for AMYR agonist (2.5 or 10 nmol / kg), and -7.1 ± 2.1 , -16.0 ± 2.5, -23.9 ± 3.5 and -23.7 ± 1.1 % of vehicle for DACRA (1 , 3, 10, or 20 nmol / kg), respectively, was observed. Body-composition measurements showed that AMYR agonist, unlike DACRA, promoted fat-mass-specific body weight loss while preserving lean-mass. The greatest fat-mass loss observed for AMYR agonist 10 nmol / kg was -28.0 ± 2.3 g, which was nearly double that observed for matched body weight loss DACRA 10 nmol / kg of -15.5 ± 6.0 g fat. Lean mass was preserved in the AMYR agonist groups, unlike in the DACRA groups (see Figure 9B). Overall, AMYR agonist resulted in more fat mass loss and preservation of lean mass as compared to DACRA at matched body weight loss levels. In particular, 100% of body weight loss was fat-mass specific in the AMYR agonist 10 nmol / kg group, as compared to 54% and 61% for DACRA 10 and 20 nmol / kg groups, respectively (see Figure 9C).

[0418] Study 2 Design: Target engagement was estimated in the following way for each compound. The average steady-state concentration (Cavg) after repeated subcutaneous injection was estimated in rats based on measured rat clearance in separate rat pharmacokinetic studies, dose level, and dosing interval. Plasma protein binding was determined in vitro using 3B Pharmaceutical’s Escalate Equilibrium Shift Assay or an in-house Surface Plasmon Resonance (SPR) assay. Peptide in vitro potency was determined via HTRF cAMP accumulation assay using HEK293 cells recombinantly expressing the rat calcitonin receptor or rat amylin receptor (calcitonin receptor plus RAMP3) in saline buffer containing 0.1% bovine serum albumin. The in vitro potencies (IC50s) for amylin and calcitonin were adjusted to unbound values by taking into account the albumin concentration in the assay and fraction unbound data, according to an established method (Wan H and Bergstrom F (2007), High Throughput Screening of Drug-Protein Binding in Drug Discovery, Journal of Liquid Chromatography & Related Technologies, 30:5, 681 -700; doi:10.1080 / 10826070701190989). Finally, the target engagement was calculated as the unbound Cavg divided by the unbound IC50, both for amylin and calcitonin. The Emax model was defined as Lean mass = E0 - Emax x (ENGAh I (ENGAh + ED50Ah)), where E0 is the baseline mass, Emax is the maximum induced mass change, ENG is the receptor engagement, and h is the slope (Hill) parameter. Parameter estimation was performed according to a maximum likelihood approach with an additive error model. Uncertainty of parameter estimates was determined by bootstrapping, sampling single measurements randomly with replacement within each experiment (N=500). Numeric analyses were performed in MATLAB (R2023b; The MathWorks, Natick, MA). The lean and fat mass changes are given as mean and standard error (N=8 per group).

[0419] Study 2 Data: Lean-mass loss correlated with the degree of calcitonin receptor engagement for DACRA, while fat-mass loss correlated with the degree of amylin receptor engagement (see Figure 9D). Thus, the AMYR agonist reduces body weight and food intake in a dosedependent manner in DIO rats and elicits fat-mass specific weight loss with preservation of lean mass.

[0420] Study 3 Design: In a separate study, male Wistar Han rats (9 weeks old) on a chow diet had overnight water restriction on day -1 . On study day 1 they were given saccharin water for 4 hours and during this time were dosed once SC. After a 4-hour exposure to saccharin, it was removed and rats had access to water as usual. A preference test was performed 72 hours after compound administration. During this time, rats are given a choice of both saccharin and water, and the intake of both are measured over a 24-hour period. Rats generally prefer the sweet saccharin water, and if they associate the saccharin water with feeling malaise or nausea they will not drink as much of the saccharin and prefer water. The saccharin preference is calculated a percent of saccharin intake (g) / total intake (saccharin and water). Data reported as mean ± standard error of the mean (SEM); data analysis performed using GraphPad Prism - statistical analysis performed using One-way ANOVA with Tukey ad hoc analysis. (*p<0.05, **p<0.01 , ***0<0.001 , ****p<0.0001 vs. Vehicle). In this experiment, the AMYR agonist was that of SEQ ID NO: 11 , and the DACRA was cagrilintide.

[0421] Study 3 Data: As shown in Figure 10A, rats treated with vehicle had a saccharin preference of >90%, while animals treated with 0.13 mg / kg DACRA had a significantly lower saccharin preference, at a similar level to 3 mg / kg cisplatin (positive control). Rats treated with 0.28 mg / kg AMYR agonist on average had a lower saccharin preference than Vehicle, albeit with a wide spread, and the average tending to lower aversion (higher saccharine preference) than the highest dose DACRA group.

[0422] The AMYR agonist exhibits ~28 fold more amylin receptor engagement than the DACRA molecule, and the DACRA exhibits ~2-3 fold more calcitonin engagement than AMYR agonist.

[0423] Study 4: The inventors discovered that saccharine aversion in lean rats correlates to calcitonin receptor engagement but not to amylin receptor engagement (see Figure 10B). Target engagement was estimated in the following way for each compound. The maximum concentration (Cmax) after subcutaneous injection was measured in separate rat pharmacokinetic studies and was dose-adjusted to the tested dose levels when appropriate, assuming dose linearity. Plasma protein binding was determined in vitro using 3B Pharmaceutical’s Escalate Equilibrium Shift Assay or an in-house Surface Plasmon Resonance (SPR) assay. Peptide in vitro potency was determined via HTRF cAMP accumulation assay using HEK293 cells recombinantly expressing the rat calcitonin receptor or rat amylin receptor (calcitonin receptor plus RAMP3) in saline buffer containing 0.1% bovine serum albumin. The in vitro potencies (IC50s) for amylin and calcitonin were adjusted to unbound values by taking into account the albumin concentration in the assay and fraction unbound data, according to an established method (Wan H and Bergstrom F (2007), High Throughput Screening of Drug-Protein Binding in Drug Discovery, Journal of Liquid Chromatography & Related Technologies, 30:5, 681 -700; doi:10.1080 / 10826070701190989). Finally, the target engagement was calculated as the unbound Cmax divided by the unbound IC50, both for amylin and calcitonin. The Emax model was defined as aversion = E0 + Emax x (ENGAh I (ENGAh + ED50Ah)), where E0 is the baseline aversion, Emax is the maximum induced aversion, ENG is the receptor engagement, and h is the slope (Hill) parameter. Parameter estimation was performed according to a maximum likelihood approach with an additive error model, incorporating data from all compounds simultaneously. Uncertainty of parameter estimates was determined by bootstrapping, sampling single measurements randomly with replacement within each experiment (N=500). Numeric analyses were performed in MATLAB (R2023b; The MathWorks, Natick, MA). The aversion data are given as mean and standard error (N=9-11 per group). In this experiment, the AMYR agonist was that of SEQ ID NO: 11 , SARA2 was eloralintide, DACRA1 was cagrilintide, and DACRA2 was petrelintide. Table 2: Relative potency ratios a: Assays in the presence of 0.1 % bovine serum albumin, n>5. b: Relative potency ratio = (amylin assay potency relative to pramlintide 100%) / (calcitonin assay potency relative to pramlintide 100%>). AMY, amylin; CTR, calcitonin, all except hAmylin and Pramlintide are lipidated peptides.

[0424] Example 5: Selective amylin receptor (AMYR) agonist monotherapy reduces body weight and food intake in a dose-dependent manner and drives fat-mass specific body weight loss and preserves lean-mass as compared to a GLP-1 R monotherapy in DIO rats

[0425] Study 1 Design: Male Sprague Dawley rats (n=5-7 per group) were used to profile a single s.c. administration of AMYR agonist or GLP-1 RA monotherapy after 6-hour fast to evaluate acute food intake in a 72-hour study. AMYR agonist doses of 3, 10, 30, 90 and 270 nmol / kg were tested, vs. 3 nmol / kg GLP-1 R agonist. In this experiment, the AMYR agonist was that of SEQ ID NO: 11 and the GLP-1 RA was semaglutide.

[0426] Study 1 Data: As shown in Figure 7, the AMYR agonist displayed dose-dependent and durable reduction in food intake in lean rats. At the highest AMYR agonist doses, food intake levels had not returned to control levels even after three dark cycles. Study 2 Design: DIO male Sprague Dawley rats (n=8 per group) were used to profile daily s.c. administration of AMYR agonist or GLP-1 RA monotherapy to evaluate body weight and body composition in a 15-day study. AMYR agonist doses of 3, 10, and 20 nmol / kg were tested, vs. 3 and 10 nmol / kg GLP-1 R agonist, and animals pair-fed to the AMYR agonist 20 nmol / kg group. In this experiment, the AMYR agonist was that of SEQ ID NO: 11 and the GLP-1 RA was semaglutide.

[0427] Study 2 Data: As shown in Figure 8A, body weight was reduced in a dose-dependent manner with repeat AMYR agonist therapy. Reduction in body weight with AMYR agonist therapy was enhanced compared to matched pair-fed animals. As shown in Figure 8B, fatmass specific body weight was reduced in a dose-dependent manner with repeat AMYR agonist therapy and enhanced compared to matched pair-fed animals, whereas lean mass was preserved. In contrast, both fat mass and lean mass were reduced following GLP-1 RA therapy. As shown in Figure 8C, at the end of the study, all AMYR agonist therapy groups displayed an increase in lean / fat mass ratio in a dose-dependent manner, with the greatest increase in the highest dose AMYR agonist therapy group.

[0428] Example 6: Selective amylin receptor (AMYR) agonist monotherapy delays gastric emptying in a dose-dependent manner in lean rats

[0429] Study Design: Lean Han Wistar rats were used to profile a single s.c. administration of AMYR agonist or vehicle to evaluate gastric emptying in a 2-hour study. AMYR agonist doses of 1 , 3 and 10 nmol / kg were tested. Barium sulphate hydroxypropyl methylcellulose (HPMC) contrast was administered 2 hours after test compound administration and gastric emptying assessed by X-ray fluoroscopy. In this experiment, the AMYR agonist was that of SEQ ID NO: 11.

[0430] Study Data: As shown in Figure 11 , the AMYR agonist therapy reduced gastric emptying in a dose-dependent manner in lean rats, demonstrating potent delayed gastric emptying with AMYR agonist administration.

Claims

1. CLAIMS1. A method of reducing the body weight of a subject, the method comprising administering to the subject about 1 mg to about 15 mg of an amylin receptor (AMYR) agonist.

2. The method of claim 1 , wherein the amount of AMYR agonist is about 1 mg to about 10 mg.

3. The method of claim 1 or claim 2, wherein the amount of AMYR agonist is about 1 mg to about 5 mg.

4. The method of claim 1 or claim 2, wherein the amount of AMYR agonist is about 4 mg to about 10 mg.

5. The method of any one of claims 1 -3, wherein the amount of AMYR agonist is about 1 mg to about 3 mg.

6. The method of any one of claims 1 -3, wherein the amount of AMYR agonist is about2.5 mg to about 5 mg.

7. The method of any one of claims 1 , 2 or 4, wherein the amount of AMYR agonist is about 7 mg to about 10 mg.

8. The method of claim 1 , wherein the amount of AMYR agonist is about 1 .5 mg, about1 .6 mg, about 4.5 mg, about 6 mg, or about 9 mg.

9. The method of any one of the preceding claims, wherein the AMYR agonist is administered by subcutaneous injection.

10. The method of any one of the preceding claims, wherein the AMYR agonist is administered to the subject by self-administration.11 . The method of any one of the preceding claims wherein the AMYR agonist is administered about once every 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, or 14 days; optionally wherein the AMYR agonist is administered about once a week.

12. The method of any one of the preceding claims, wherein the AMYR agonist is administered to the subject multiple times over a period of at least 3 months, 6 months, 9 months, 1 year, 2 years, or 5 years.

13. The method of any one of the preceding claims, wherein the AMYR agonist is titrated to a final dose over a period between about 4 weeks and about 8 weeks.

14. The method of any one of the preceding claims, wherein the subject’s body weight is reduced by >5%, optionally, wherein the subject’s body weight is reduced by >10%, >15%, or >20%.

15. The method of any one of the preceding claims, wherein the subject’s reduction in body weight is fat-specific weight loss.

16. The method of any one of the preceding claims, wherein the subject experiences reduced nausea as compared to a therapeutically effective dose of a dual amylin- calcitonin receptor agonist (DACRA) or a GLP-1 R agonist.

17. The method of any one of the preceding claims, wherein the AMYR agonist has selectivity to AMYR as compared to a calcitonin receptor (CTR).

18. The method of claim 17, wherein the AMYR agonist has at least a 2-fold, at least 3- fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 12-fold, at least 15-fold, at least 17-fold, at least 20-fold, or at least 25-fold selectivity to AMYR as compared to CTR; optionally wherein the AMYR agonist has at least a 10-fold selectivity to AMYR as compared to CTR.

19. The method of any one of the preceding claims, wherein the AMYR agonist is a polypeptide, small molecule drug, antibody, antibody-drug conjugate, or aptamer; or a pharmaceutically acceptable salt thereof.

20. The method of claim 19, wherein the AMYR agonist is a polypeptide, or a pharmaceutically acceptable salt thereof.21 . The method of any one of the preceding claims, wherein the AMYR is a human AMYR; and / or wherein the AMYR is AMY1 R, AMY2R and / or AMY3R.

22. The method of any one of the preceding claims, wherein the AMYR agonist is an AMYR agonist polypeptide, or a pharmaceutically acceptable salt thereof, which comprises an amino acid sequence having at least 90% identity to pramlintide (KCNTATCATQRLANFLVHSSNNFGPILPPTNVGSNTY-amide, SEQ ID NO: 5).

23. The method of claim 22, wherein the AMYR agonist polypeptide is lipidated and / or wherein the lipid is attached to an amino acid residue in the AMYR agonist polypeptide by a linker.

24. The method of claim 22 or claim 23, wherein the AMYR agonist polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 99%, or 100% identity to the amino acid sequenceK[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 26).

25. The method of claim 24, wherein the AMYR agonist polypeptide, or pharmaceutically acceptable salt thereof, comprises the amino acid sequence K(yE-yE- C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 11 ).

26. The method of any one of the preceding claims, wherein the subject has a disease or disorder selected from obesity, metabolic disease, an obesity-related condition, eating disorder, Alzheimer's disease, hepatic steatosis ("fatty liver"), kidney failure, arteriosclerosis (e.g. atherosclerosis), cardiovascular disease, macrovascular disease, microvascular disease, diabetic heart (including diabetic cardiomyopathy and heart failure as a diabetic complication), coronary heart disease, peripheral artery disease or stroke, cancer, dumping syndrome, hypertension e.g. pulmonary hypertension, or dyslipidemia e.g. atherogenic dyslipidemia, cholescystitis, or short bowel syndrome.

27. The method of claim 26, wherein the subject has an obesity-related condition; optionally wherein the obesity-related condition is overweight, morbid obesity, obesity prior to surgery, obesity-linked inflammation, obesity-linked gallbladder disease, sleep apnoea and respiratory problems, hyperlipidaemia, degeneration of cartilage, osteoarthritis, or reproductive health complications of obesity or overweight such as infertility; optionally wherein the subject is overweight; further optionally wherein the subject is overweight with at least one comorbidity selected from hypertension, dyslipidaemia, cardiovascular (CV) disease, or obstructive sleep apnoea.

28. The method of claim 26, wherein the subject has a metabolic disease, optionally wherein the metabolic disease includes diabetes, type 1 diabetes, type 2 diabetes, gestational diabetes, pre-diabetes, insulin resistance, impaired glucose tolerance(IGI), disease states associated with elevated blood glucose levels, metabolic syndrome, or hyperglycaemia (e.g. abnormal postprandial hyperglycaemia).

29. The method of any one of the preceding claims, wherein the AMYR agonist is a polypeptide, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence K(yE-yE-C18diacid)[CNTATC]ATQRLANFLRHSSNN(aMePhe)GPILPPTEVGSNTY-amide (SEQ ID NO: 11 ) in an amount of about 1 .5 mg, about 1 .6 mg, about 4.5 mg, about 6 mg, or about 9 mg, and wherein the subject has obesity or an obesity-related condition.

30. A method of treating and / or preventing obesity or an obesity-related condition in a subject in need thereof, the method comprising administering to the subject about 1 mg to about 15 mg of an amylin receptor (AMYR) agonist.31 . A method of treating and / or preventing a metabolic disease in a subject in need thereof, the method comprising administering to the subject about 1 mg to about 15 mg of an amylin receptor (AMYR) agonist.

32. A cosmetic method of reducing the body weight of a subject, the method comprising administering to the subject about 1 mg to about 15 mg of an amylin receptor (AMYR) agonist.

33. An AMYR agonist for use in a method of treating and / or preventing obesity or an obesity-related condition in a subject in need thereof, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist.

34. An AMYR agonist for use in a method of treating and / or preventing a metabolic disease in a subject in need thereof, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist.

35. An AMYR agonist for use in a method of reducing body weight in a subject, the method comprising administering to the subject about 1 mg to about 15 mg of the AMYR agonist.

36. The AMYR agonist for use in a method according to claim 35, wherein the subject has a disease or disorder selected from obesity, an obesity-related condition, and metabolic disease.

7. A kit comprising (a) an AMYR agonist, and (b) instructions for use of the same for treating or preventing obesity or an obesity-related condition at a dose of about 1 mg to about 15 mg.

Images