Analogues with improved properties

Abstract

The present invention relates to therapeutic and non-therapeutic methods of using peptide hormone analogs. In particular, the present invention relates to the use of amylin analogs with improved properties, in particular amylin analogs that do not delay gastric emptying.

Description

TECHNICAL FIELD

[0001] The present invention relates to therapeutic and non-therapeutic methods using peptide hormone analogues. In particular, the present invention relates to the use of amylin analogues having improved properties, in particular amylin analogues that do not delay gastric emptying. BACKGROUND

[0002] Obesity is a major public health problem in many developed countries today and is associated with the development of several serious conditions such as cardiovascular disease, type 2 diabetes, sleep apnoea and certain cancers. The standard treatment for obesity is lifestyle intervention, including reducing energy intake and increasing exercise. However, while such interventions can achieve temporary success, it is often challenging for patients to maintain such lifestyle changes over a prolonged period of time such that permanent weight loss is achieved. Therefore, more effective pharmacological treatment options are needed.

[0003] Amylin is a peptide hormone family that includes amylin, calcitonin, calcitonin gene-related peptide, adrenomedullin and midkine (midkine is also known as AFP-6) and has been implicated in a variety of metabolic diseases and conditions.

[0004] Human amylin was first isolated, purified and characterised as a major component of amyloid deposits in the islets of Langerhans from the pancreas of patients with type 2 diabetes.

[0005] Native human amylin is a 37 amino acid peptide having the following formula:

[0006]

[0007] where H- at the N-terminus represents a hydrogen atom corresponding to the presence of a free amino group on the N-terminal amino acid residue [i.e. the lysine (K) residue at position 1 in the sequence shown above]; where -NH2 at the C-terminus represents that the carboxyl group at the C-terminus is in amide form; and where the parentheses "()" linked to the two cysteine (C, Cys) residues at positions 2 and 7 indicate the presence of an intramolecular disulphide bridge between the two Cys residues involved.

[0008] Amylin can be beneficial in the treatment of metabolic conditions such as diabetes and / or obesity. Amylin is thought to slow gastric emptying, suppress glucagon secretion and reduce food intake, thereby modulating the rate of glucose release into the circulation. Amylin appears to complement the action of insulin. Type 1 diabetic patients do not have circulating amylin and type 2 diabetic patients show reduced postprandial amylin concentrations compared to healthy adults. WO 93 / 10146 describes an amylin analogue known as pramlintide, which has the following sequence:

[0009] .

[0010] Pramlintide also has a disulfide bridge between the cysteine residues at position 2 and 7 and has shown to reduce body weight or reduce body weight gain in human trials.

[0011] Yan et al. (PNAS, 103(7), 2046-2051, 2006; Angew. Chem. Int. Ed. 2013, 52, 10378-10383; WO2006 / 042745) have described an alternative amylin analogue incorporating N-methylated residues and having a reduced fibrillation propensity, which is designated IAPP-GI. However, IAPP-GI shows lower activity than native amylin.

[0012] WO 2018 / 046719 describes amylin analogues having, inter alia, a lactam bridge instead of a disulfide bridge, having N-methylated residues, and having a deletion of residues Asn21 and Asn22 corresponding to native human amylin. Such analogues have a significantly lower fibrillation propensity than native amylin, while also having a higher potency than the analogue described by Yan et al. (supra).

[0013] While the delayed gastric emptying associated with amylin and amylin analogues can result in a beneficial reduction of food intake, delayed gastric emptying can in some cases be undesirable.

[0014] For example, delayed gastric emptying can result in unwanted changes to exposure to or need for concomitant drugs. Such drug-drug interactions are undesirable. SUMMARY

[0015] The present invention relates to the surprising finding that amylin analogues induce body weight loss, reduce glucagon release and increase satiety in a subject, while unexpectedly not affecting gastric emptying.

[0016] The present invention thus relates to functional analogues of amylin that do not cause delayed gastric emptying.

[0017] In one general aspect, the present invention relates to a dosing regimen for a subject, wherein the dosing regimen comprises:

[0018] (a) administering to the subject an amylin analogue that does not affect the rate of gastric emptying; and

[0019] (b) administering to the subject a concomitant agent.

[0020] According to the present application, the amylin analogue which does not affect the rate of gastric emptying comprises or consists of a peptide of the formula:

[0021]

[0022] wherein:

[0023] R 1 is hydrogen, C 1-4 1-4alkyl, acyl, benzoyl or C 1-4 1-4alkyl, or a half-life extending moiety M, wherein M is optionally linked to Z via a linker moiety L;

[0024] R 2 is OH or NHR 3 , wherein R 3 is hydrogen or C 1-3 1-4alkyl; and

[0025] Z is an amino acid sequence of the formula I:

[0026]

[0027] wherein:

[0028] X1is selected from Arg, Lys and Glu;

[0029] X3is selected from Gly, Gin and Pro;

[0030] X4is selected from Thr and Glu;

[0031] X5is selected from Ala and Leu;

[0032] X6is selected from Thr and Ser;

[0033] X10is selected from Glu and Gin;

[0034] X14is selected from Aad, His, Asp, Asn and Arg;

[0035] X17is selected from Gin, His and Thr;

[0036] X19-X20is selected from Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly and Ala-Asn, or is absent;

[0037] X27is selected from Leu and Pro;

[0038] X32is selected from Val and Thr;

[0039] X35is selected from Asn and Ser;

[0040] X37 is selected from Hyp and Pro; and

[0041] X2 and X7 are amino acid residues that together form a lactam bridge in their side chains;

[0042] Gly(Me) is N-methylglycine [also known as sarcosine (Sar)].

[0043] Ile(Me) is N-methylisoleucine

[0044] Aad stands for 2-aminoadipic acid, such as (2S)-2-aminoadipic acid [also known as (2S)-2-aminoadipic acid], or homo-glutamic acid.

[0045] Preferably, the amylin analogue comprises a peptide of the following formula or a pharmaceutically acceptable salt and / or derivative thereof, or is composed of a peptide of the following formula or a pharmaceutically acceptable salt and / or derivative thereof:

[0046] .

[0047] In some preferred embodiments, the amylin analogue is petrelintide, which is a compound having the following formula or a pharmaceutically acceptable salt thereof:

[0048]

[0049] Intramolecular lactam bridges are formed between the side chains of the residues indicated by parentheses “()”; and [19CD]-isoGlu is a 19-carboxynonadecanoyl group covalently linked to the α-amino group of the isoglutamic acid linker.

[0050] In one aspect, the present invention provides a dosing regimen for a subject, wherein the dosing regimen comprises:

[0051] (a) Administration of amylin analogues to a subject that do not affect the rate of gastric emptying; and

[0052] (b) Applying a comorbid agent to the object;

[0053] The amylin analogue and its concomitant are administered to the subject within approximately 120 minutes of each other.

[0054] In one aspect, the present invention provides a method for inhibiting or reducing weight gain, promoting weight loss, reducing food intake, and / or losing weight in a subject, wherein the method comprises:

[0055] (a) Administration of amylin analogues to a subject that do not affect the rate of gastric emptying; and

[0056] (b) administering to the subject an accompaniment;

[0057] wherein the amylin analogue and the accompaniment are administered to the subject within about 120 minutes of each other.

[0058] In one aspect, the method of inhibiting or reducing weight gain, promoting weight loss, reducing food intake, and / or reducing excess weight in a subject is a non-therapeutic method.

[0059] In one aspect, the present application provides a method of treating or preventing a disease in a subject, wherein the method comprises:

[0060] (a) administering to the subject an amylin analogue that does not affect gastric emptying rate; and

[0061] (b) administering to the subject an accompaniment;

[0062] wherein the amylin analogue and the accompaniment are administered to the subject within about 120 minutes of each other.

[0063] In one aspect, the present application provides an amylin analogue that does not affect gastric emptying rate for use in a method of treating or preventing a disease in a subject, wherein the method comprises:

[0064] (a) administering to the subject an amylin analogue; and

[0065] (b) administering to the subject an accompaniment;

[0066] wherein the amylin analogue and the accompaniment are administered to the subject within about 120 minutes of each other.

[0067] In one aspect, the present application provides the use of an amylin analogue that does not affect gastric emptying rate for the manufacture of a medicament for use in a method of treating or preventing a disease in a subject, wherein the method comprises:

[0068] (a) administering to the subject an amylin analogue; and

[0069] (b) administering to the subject an accompaniment;

[0070] wherein the amylin analogue and the accompaniment are administered to the subject within about 120 minutes of each other. BRIEF DESCRIPTION OF DRAWINGS

[0071] Figure 1 : Single-dose escalation trial design

[0072] The single ascending dose (SAD) trial of Example 1 comprised seven cohorts, each consisting of six participants receiving the amylin analogue of SEQ ID NO: 3 and two participants receiving a placebo. The dose was administered subcutaneously. An additional cohort of the same composition (not shown) received an intravenous dose of 0.35 mg.

[0073] Figure 2 The pharmacokinetic profiles of the groups are consistent.

[0074] The pharmacokinetic profiles for each group are shown. Individual blood concentrations of amylin analogues after a single subcutaneous dose are also shown. Figure 2 A), and each group lasted for up to 35 days ( Figure 2 B) and lasting up to 7 days ( Figure 2 The geometric mean within C) is shown. Each group exhibits a consistent pharmacokinetic profile. Furthermore, exposure variability within the groups is small.

[0075] Figure 3 Dose-dependent reduction in body weight

[0076] A dose-dependent and sustained decrease in body weight was observed in subjects who received a single dose of an amylin analogue. Figure 3 A) shows the weight changes of individual subjects in each group. Figure 3 B) shows the average change in body weight and the 95% confidence level.

[0077] Figure 4 A dose-dependent decrease in glucagon secretion

[0078] A dose-dependent decrease in glucagon secretion was observed in subjects who received a single dose of an amylin analogue following a mixed test meal (MTM).

[0079] Figure 5 Acetaminophen absorption is not affected by amylin analogues.

[0080] No dose-response relationship was observed between amylin analogues and acetaminophen absorption. Amylin analogues substantially did not alter blood acetaminophen concentrations measured after the mixed test diet (MTM) and acetaminophen intake. Figure 5 In A), the concentration of acetaminophen in the blood is shown as a function of time after MTM; the area under the curve corresponding to the first hour after MTM is also shown. Figure 5 B). Figure 5C) Gastric emptying (AUC of acetaminophen concentration) shown as a percentage of placebo further indicates no significant effect on acetaminophen concentration and thus no significant effect on gastric emptying.

[0081] Figure 6 Multiple ascending dose trial design

[0082] The Multiple Ascending Dose (MAD) trial of Example 2 included 2 cohorts, each consisting of 7 participants receiving the amylin analogue of SEQ ID NO: 3 and 3 participants receiving placebo. Doses were administered subcutaneously.

[0083] Figure 7 Weight loss

[0084] Weight loss was observed in human subjects receiving multiple doses of the amylin analogue. The graphs (from left to right) are placebo, amylin 0.6 mg, and amylin 1.2 mg. The graphs show individual subjects (thin lines) and mean values (thick lines) for each cohort.

[0085] Figure 8 Absorption of acetaminophen is not affected by amylin analogue

[0086] No dose relationship was observed between the amylin analogue and acetaminophen absorption. Blood acetaminophen concentrations following mixed test meal (MTM) and acetaminophen intake measured on days 5 and 40 were essentially unchanged from day -1 by the amylin analogue.

[0087] Figure 9 Decreased glucagon secretion

[0088] Decreased glucagon secretion following a mixed test meal (MTM) on day 5 was observed in participants receiving doses of the amylin analogue.

[0089] Figure 10 Multiple ascending dose trial design - Part 2

[0090] The Multiple Ascending Dose (MAD) trial of Example 3 included 3 cohorts, each consisting of 12 participants receiving the amylin analogue of SEQ ID NO: 3 and 4 participants receiving placebo. Doses were administered subcutaneously. DETAILED DESCRIPTION

[0091] Unless otherwise defined herein, scientific and technical terms used in this application have the meanings that are commonly understood by one of ordinary skill in the art. Generally, nomenclature used in connection with, and techniques related to, chemical, molecular biology, cell and cancer biology, immunology, microbiology, pharmacology, and protein and nucleic acid chemistry described herein are those well-known and commonly used in the art.

[0092] All patents, published patent applications, and non-patent publications referred to in this application are specifically incorporated by reference herein. In the event that any inconsistency or conflict exists among cited references, the present specification, including definitions, controls.

[0093] Each embodiment of the application described herein can be used alone or in combination with one or more other embodiments of the application.

[0094] Various modifications and variations of the described methods and systems of the application will be apparent to those skilled in the art without departing from the scope and spirit of the application.

[0095] While the application has been described in connection with specific preferred embodiments thereof, it will be understood that the application is not limited to these specific embodiments. Indeed, various modifications of the described modes of carrying out the application that are obvious to those skilled in biochemistry, molecular biology or related fields are intended to be within the scope of the following.

[0096] Definitions

[0097] The following definitions are provided for specific terms used herein in the written description. All other terms are to be understood as having their common meaning as known to one of skill in the art unless otherwise specified.

[0098] Throughout this specification the word "comprise," or variations such as "comprises" or "comprising," will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.

[0099] Unless otherwise indicated, a noun clause of the singular form is intended to denote one or more.

[0100] The term "including" is used to mean "including but not limited to." "Including" and "including but not limited to" are used interchangeably.

[0101] The terms "patient," "subject," and "individual" are used interchangeably and refer to either a human or a non-human animal. These terms include mammals, such as humans, primates, livestock animals (e.g., cows and pigs), companion animals (e.g., canids and felines), and rodents (e.g., mice and rats).

[0102] The term "solvate" in the context of the present application refers to a complex of defined stoichiometry between a solute (in this case a peptide or a pharmaceutically acceptable salt thereof according to the present application) and a solvent. In this case, the solvent can be, for example, water, ethanol or other pharmaceutically acceptable, usually small-molecular organic substances such as, but not limited to, acetic acid or lactic acid. When the solvent involved is water, such solvates are commonly referred to as hydrates.

[0103] Sequence alignments can be performed by the skilled person using techniques well known in the art, for example using publicly available software such as BLAST, BLAST2 or Align software. See, for example, Altschul et al., Methods in Enzymology 266: 460-480 (1996) or Pearson et al., Genomics 46: 24-36, 1997.

[0104] Percent sequence identity as used herein can be determined using these programs with their default settings in the context of the present application. More generally, the skilled person can readily determine suitable parameters for determining alignments, including any algorithms required to achieve maximum alignment over the full length of the sequences being compared.

[0105] Amylin analog

[0106] An amylin analog is a molecule that acts as an amylin receptor agonist, i.e. the molecule is capable of binding to and inducing signalling by one or more receptors or receptor complexes that are considered to be physiological receptors for human amylin. The terms "amylin analog" and "amylin receptor agonist" are used interchangeably herein. An amylin analog can also be referred to herein as a compound, molecule or peptide.

[0107] An amylin analog (which can also be referred to herein as a compound, molecule or peptide) can be suitably prepared by standard synthetic methods. Thus, a peptide can be synthesized by, for example, a method comprising synthesizing the peptide stepwise or by fragment assembly by standard solid phase or liquid phase methods, and optionally isolating and purifying the final peptide product. The method will typically also comprise a step of forming an amide bond between the side chains at positions 2 and 7, for example as described below. In the case of solid phase synthesis, cyclization can be performed in situ on the solid phase (e.g. resin), i.e. before the peptide is removed from the solid phase.

[0108] The amylin analogs described herein can have the following formula or be a pharmaceutically acceptable salt or solvate thereof:

[0109]

[0110] wherein:

[0111] R 1 is hydrogen, C 1-4 alkyl, acyl, benzoyl or C 1-4 alkyl, or a half-life extending moiety M, wherein M is optionally linked to Z through a linker moiety L;

[0112] R 2 is OH or NHR 3 , wherein R 3 is hydrogen or C 1-3 alkyl; and

[0113] Z is an amino acid sequence of Formula I:

[0114]

[0115] and wherein:

[0116] X1is selected from Arg, Lys and Glu;

[0117] X3is selected from Gly, Gin and Pro;

[0118] X4is selected from Thr and Glu;

[0119] X5is selected from Ala and Leu;

[0120] X6is selected from Thr and Ser;

[0121] X10is selected from Glu and Gin;

[0122] X14is selected from Aad, His, Asp, Asn and Arg;

[0123] X17is selected from Gin, His and Thr;

[0124] X19-X20is selected from Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly and Ala-Asn, or is absent;

[0125] X27 is selected from Leu and Pro;

[0126] X32 is selected from Val and Thr;

[0127] X35 is selected from Asn and Ser;

[0128] X37 is selected from Hyp and Pro; and

[0129] X2 and X7 are amino acid residues whose side chains together form a lactam bridge;

[0130] Gly(Me): N-methylglycine [also known as sarcosine (Sar)]

[0131] Ile(Me): N-methylisoleucine

[0132] Aad: 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2- amino hexanedioic acid], also known as homoglutamic acid

[0133] Preferably, the amylin analogue is petrelintide, which has the following formula:

[0134]

[0135] wherein:

[0136] Gly(Me): N-methylglycine [also known as sarcosine (Sar)]

[0137] Ile(Me): N-methylisoleucine

[0138] Aad: 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2- amino hexanedioic acid], also known as homoglutamic acid

[0139] Hyp: 4-hydroxyproline, for example (2S,4R)-4-hydroxyproline [also known as (4R)-4- hydroxy-L-proline]

[0140] [19CD]-isoGlu: the backbone nitrogen of the Arg residue (present at position X1 of the peptide sequence Z of the amylin analogue) is covalently linked via an amide linkage to the side chain carboxyl group of the Glu moiety. The 19-carboxy-nonadecanoyl group is covalently linked via an amide linkage to the alpha amino group of the Glu linker (as described below).

[0141] The brackets "()" shown after a particular amino acid residue symbol indicate that the side chain of this residue participates in an intramolecular lactam bridge. Thus, the amylin analogue compounds of the present application have an intramolecular lactam bridge formed between the side chains of the residues indicated by the brackets (aspartic acid and lysine, respectively).

[0142] In some preferred embodiments, the amylin analogue is petrelintide, which is a compound having the following formula:

[0143]

[0144] wherein an intramolecular lactam bridge is formed between the side chains of the residues indicated by the brackets; and wherein [19CD]-isoGlu is a 19-carboxy-nonadecanoyl group covalently linked to the alpha amino group of the isoGlu linker.

[0145] Throughout the present specification and claims, the conventional three-letter and one-letter codes for naturally occurring amino acids are used, i.e.:

[0146]

[0147] and the generally accepted three-letter codes for other alpha-amino acids, such as sarcosine (Sar), norleucine (Nle), alpha-aminoisobutyric acid (Aib), 2,3-diaminopropionic acid (Dap), 2,4-diaminobutyric acid (Dab) and 2,5-diaminopentanoic acid (ornithine; Orn). Such other alpha-amino acids can be shown in square brackets "[ ]" (e.g. "[Aib]") when used in a general formula or sequence in the present specification, especially when the remainder of the formula or sequence is shown using one-letter codes. Unless otherwise indicated, the amino acid residues in the peptides of the present application are in the L-configuration. However, D-configuration amino acids can be incorporated. In the context of the present application, an amino acid code written in lower case letters represents the D-configuration of the amino acid, e.g. "k" represents the D-configuration of lysine (K).

[0148] The amylin analogue of SEQ ID NO: 3 is petrelintide and can also be referred to as ZP8396. ZP8396 (petrelintide) is also described in detail in WO 2018 / 046719, which is incorporated herein by reference in its entirety. In WO 2018 / 046719, the amylin analogue of SEQ ID NO: 3 (ZP8396, petrelintide) is also referred to as "Compound 35".

[0149] The amylin analogue can be in the form of a pharmaceutically acceptable salt or solvate, for example a pharmaceutically acceptable acid addition salt.

[0150] Half-life extending moiety M

[0151] The N-terminal moiety R in the compounds of the application 1 may be a half-life extending moiety M (sometimes referred to in the literature as a duration enhancing moiety or an albumin binding moiety) which is optionally linked (covalently linked) to the peptide moiety Z by a linker moiety L. Suitable half-life extending moieties include certain types of lipophilic substituents. Without wishing to be bound by any particular theory, it is believed that such lipophilic substituents (as well as other kinds of half-life extending moieties) bind to albumin in the blood stream, thereby protecting the compounds of the application from filtration by the kidneys as well as from enzymatic degradation, and thus can increase the half-life of the compounds in the body. The lipophilic substituents can also modulate the potency of the compounds as amylin (calcitonin) receptor agonists.

[0152] The lipophilic substituent can be linked to the N-terminal amino acid residue or to the linker L by an ester, a sulfonyl ester, a thioester, an amide, an amine or a sulfonamide. Thus, it will be appreciated that preferably the lipophilic substituent comprises an acyl group, a sulfonyl group, an N atom, an O atom or an S atom which forms part of the ester, sulfonyl ester, thioester, amide, amine or sulfonamide. Preferably, the acyl group in the lipophilic substituent forms part of an amide or ester with the amino acid residue or linker.

[0153] The lipophilic substituent can comprise a hydrocarbon chain having 10 to 24 C atoms (e.g. 14 to 22 C atoms, e.g. 16 to 20 C atoms). Preferably it has at least 14 C atoms and preferably has 20 C atoms or less. For example, the hydrocarbon chain can contain 14, 15, 16, 17, 18, 19 or 20 carbon atoms. The hydrocarbon chain can be linear or branched and can be saturated or unsaturated. Furthermore, it can comprise a functional group at the end of the hydrocarbon chain, for example a carboxylic acid group, which can or can not be protected during synthesis. It will also be appreciated from the above discussion that the hydrocarbon chain is preferably substituted with a moiety which forms part of the linkage to the N-terminal amino acid residue of the peptide moiety Z or to the linker L, e.g. an acyl group, a sulfonyl group, an N atom, an O atom or an S atom.

[0154] Most preferably, the hydrocarbon chain is substituted with an acyl group and thus the hydrocarbon chain can be part of an alkanoyl group, such as dodecanoyl, 2-butyloctanoyl, tetradecanoyl, hexadecanoyl, heptadecanoyl, octadecanoyl, nonadecanoyl or icosanoyl. Some examples of functionalized hydrocarbon chains are 15-carboxy-pentadecanoyl, 17-carboxy-heptadecanoyl and 19-carboxy-nonadecanoyl.

[0155] As mentioned above, the lipophilic substituent M can be connected to the N-terminal amino acid residue of Z via a linker L. In some embodiments, the linker moiety L can itself comprise one, two, three or more connected submoieties L 1 , L 2 , L 3 etc. When the linker L comprises only one such moiety, it is connected to the lipophilic substituent as well as to the N-terminal amino acid residue of Z. The linker can then independently be connected to the lipophilic substituent as well as to the N-terminal amino acid residue of Z via an ester, a sulfonyl ester, a thioester, an amide, an amine or a sulfonamide linkage. It can thus comprise two moieties independently selected from acyl, sulfonyl, N atoms, O atoms and S atoms. The linker can consist of a straight-chain C 1-10 hydrocarbon chain or a branched-chain C 1-10 hydrocarbon chain or more preferably of a straight-chain C 1-5 hydrocarbon chain. Furthermore, the linker can be substituted with one or more substituents selected from C 1-6 alkyl, amino C 1-6 alkyl, hydroxy C 1-6 alkyl and carboxy C 1-6 alkyl.

[0156] In some embodiments, the linker can comprise one or more (e.g. one, two or three) connected amino acid residues, which can each independently be a residue of any naturally occurring amino acid or a non-naturally occurring amino acid. For example, the linker can comprise one, two or three connected amino acid residues, each of which can independently be a residue of:

[0157]

[0158] (i.e. 3-aminopropanoyl), 4-aminobutanoic, 5-aminopentanoyl, 6-aminohexanoyl, 7- aminohexanoyl, 8-aminooctanoyl, 9- aminononanoyl, 10-aminodecanoyl or 8Ado (i.e. 8-amino-3,6-dioxaoctanoyl).

[0159] References to γ-Glu, ε-Lys and β-Asp denote the residue of an amino acid which participates in a bond via its side chain carboxyl or amine function. Thus γ-Glu and β-Asp participate in a bond via their alpha amino group and side chain carboxyl, while ε-Lys participates via its carboxyl and side chain amino group. In the context of the present invention, γ-Glu and isoGlu are used interchangeably.

[0160] In certain embodiments, the linker comprises or consists of one, two or three residues of independently selected Glu, γ-Glu, ε-Lys, β-Ala, 4-aminobutyryl, 8-aminooctanoyl or 8Ado.

[0161] A linker consisting of isoGlu and isoGlu-isoGlu can be particularly preferred.

[0162] One example of a lipophilic substituent comprising a lipophilic moiety M and a linker L is shown in the following formula:

[0163] .

[0164] Here the backbone nitrogen of the Arg residue (present in position X1 of the peptide sequence Z of the amylin analogue) is covalently linked via an amide linkage to the side chain carboxyl of the Glu moiety. The 19-carboxy-nonadecanoyl group is covalently linked via an amide linkage to the alpha amino group of the Glu linker. The Glu linker is thus in iso-Glu (or γ-Glu) configuration. This combination of lipophilic moiety and linker attached to the Arg residue can be represented by the shorthand notation [19CD]-isoGlu-R, for example when shown in a formula of a particular compound.

[0165] The skilled person will be well aware of suitable techniques for preparing the compounds employed in the context of the present invention. Examples of suitable chemical procedures are found in, for example, WO 98 / 08871, WO 00 / 55184, WO 00 / 55119, Madsen et al. (J. Med. Chem. 2007, 50, 6126-32) and Knudsen et al. 2000 (J. Med Chem. 43, 1664-1669).

[0166] The hydrocarbon chain in the lipophilic substituent can be further substituted. For example, it can also be substituted with up to three substituents selected from NH2, OH and COOH. If the hydrocarbon chain is further substituted, it is preferred that it is further substituted with only one substituent. Alternatively or additionally, the hydrocarbon chain can include a cycloalkane or heterocycloalkane moiety, for example as shown below:

[0167] .

[0168] In some embodiments, the cycloalkane or heterocycloalkane moiety is a six-membered ring, such as a piperidine ring.

[0169] In some alternative embodiments of the application, the N-terminal amino acid of Z in the compounds of the application can be optionally linked (covalently linked) to a biotinylated substituent via a linker moiety L. Without wishing to be bound by any particular theory, it is believed that such a biotinylated substituent binds to albumin in the bloodstream, thereby protecting the compound of the application from enzymatic degradation, and thus can increase the half-life of the compound in vivo. When a linker is present, it can provide a spacer between the peptide moiety Z and the biotinylated substituent.

[0170] The biotinylated substituent can be linked to the N-terminal amino acid residue or to the linker via a maleimide ester bond, a sulfonyl ester bond, a thioester bond, an amide bond, an amine bond, or a sulfonamide bond. Thus, it will be appreciated that the biotinylated substituent preferably comprises a maleimide group, an acyl group, a sulfonyl group, an N atom, an O atom, or an S atom that forms part of the ester, sulfonyl ester, thioester, amide, amine, or sulfonamide bond involved.

[0171] Some examples of biotinylated substituents can include

[0172] .

[0173] Biotin is known as vitamin H or coenzyme R, and is a water-soluble B-complex vitamin (vitamin B7). It has been shown to improve oral uptake of certain drugs.

[0174] Potency of compounds

[0175] The compounds of the application, i.e. the amylin analogs, are amylin receptor agonists, i.e. they are capable of binding to and inducing signaling through one or more of the receptors or receptor complexes that are considered to be the physiological receptors of human amylin. These include the human calcitonin receptor hCT-R, and complexes comprising the human calcitonin receptor hCT-R with at least one of the human receptor activity modifying proteins designated hRAMP1, hRAMP2 and hRAMP3. The complexes between hCT-R and hRAMP1, hRAMP2 and hRAMP3 are designated hAMYR1, hAMYR2 and hAMYR3, respectively (i.e. human amylin receptors 1, 2 and 3).

[0176] A compound can be considered an amylin receptor agonist if it has agonist activity on one or more of hAMYR1, hAMYR2 and hAMYR3 (e.g. on hAMYR1 and / or hAMYR3, e.g. on hAMYR3).

[0177] Generally, the amylin receptor agonist will also have agonist activity at the hCT-R when expressed in the absence of hRAMP1, hRAMP2 and hRAMP3. Generally, the agonist activity at the hCT-R (when expressed in the absence of hRAMP1, hRAMP2 and hRAMP3) is less than 10-fold higher than its activity at any one of hAMYR1, hAMYR2 and hAMYR3 (i.e. its activity at all of these receptors) in a comparable assay. The agonist activity at the hCT-R can be less than 5-fold higher than the agonist activity at hAMYR1, hAMYR2 and hAMYR3, substantially the same as (e.g. + / - 10%) the agonist activity at hAMYR1, hAMYR2 and hAMYR3, or less than the agonist activity at hAMYR1, hAMYR2 and hAMYR3. In this regard, it is sufficient to compare activity between the hCT-R and hAMYR3 only.

[0178] The ability to induce cAMP formation (i.e. to induce adenylyl cyclase activity) as a result of binding to the relevant receptor or receptor complex is generally considered to be an indication of agonist activity. Other intracellular signalling pathways or events can also be used as read-outs of amylin receptor agonist activity. These can include calcium release, betacellulin recruitment, receptor internalisation, kinase activation or inactivation, lipase activation, inositol phosphate release, diacylglycerol release or nuclear transcription factor translocation.

[0179] A suitable comparable assay format will use cells expressing hCT-R and differ only in their expression of hRAMP1, 2 and 3. For example, a "base" cell line which does not express any of hCT-R, hRAMP1, hRAMP2 and hRAMP3 can be engineered to produce cells which express (i) hCT-R and (ii) one of hAMYR1, hAMYR2 and hAMYR3 (i.e. hCT-R plus one of hRAMP1, hRAMP2 and hRAMP3), for example hAMYR3. The base cells are typically mammalian cells and can be primate cells. They can be non-human primate cells. Preferably, the base cells do not express any of CT-R, RAMP1, RAMP2 or RAMP3 (whether human or native to the base cells if the base cells are non-human). The base cells can be fibroblasts. Suitable non-human fibroblast base cells include COS7 cells from African green monkey which do not express native CT-R or RAMPs.

[0180] Comparable activity can be measured by any suitable means, for example by EC 50The determination of the value is measured. It is clear that the biological readout must be the same for both receptor types.

[0181] The compounds of the present application can exhibit a number of advantageous properties relative to human amylin and its existing analogs (e.g. pramlintide, IAPP-GI, and the analogs described in WO2012 / 168430, WO2012 / 168431 and WO2012 / 168432). The compounds of the present application can exhibit, for example, improved potency (e.g. in the form of improved in vitro activity or potency at one or more of the receptors hCT-R, hAMYR1, hAMYR2 or hAMYR3) compared to human amylin or any of these analogs. In addition or alternatively, the compounds of the present application can exhibit improved solubility in aqueous media, especially at a pH of 4 to 7.5, or a pH range within this range. Furthermore, the compounds of the present application can exhibit, in addition or alternatively, a reduced tendency to fibrillate in pharmaceutically relevant aqueous media, especially at a pH of 4 to 7, or a pH range within this range. Furthermore, the compounds of the present application can exhibit, in addition or alternatively, improved chemical stability (i.e. a reduced tendency to chemically degrade) in aqueous media, especially at a pH of 4 to 9, or a pH range within this range.

[0182] Thus, the compounds of the present application can be very well suited for formulation in acidic media (e.g. pH 4) and in neutral or near neutral media (e.g. pH 7 or 7.4). This is in contrast to, for example, pramlintide which typically exhibits poor chemical stability and rapid fibrillation in pharmaceutically relevant aqueous media at neutral pH. Thus, the compounds of the present application can be very well suited for co-formulation with, for example, insulin, various insulin analogs and / or other therapeutic (e.g. anti-diabetic or anti-obesity) agents which require a neutral or near neutral formulation pH.

[0183] Surprisingly, it has been found that the compounds of the present application (amylin analogs) do not induce gastric emptying delay. Thus, these compounds (amylin analogs) do not alter (e.g. do not slow down) gastric emptying when administered to a subject.

[0184] In general, it is preferred to use a biological assay that measures the intracellular signaling caused by binding of a compound to the relevant receptor as discussed above. Activation of the calcitonin / amylin receptor by a compound of the present application, which acts as an agonist of the receptor, induces cAMP formation and activates other intracellular signaling pathways and events. Thus, in suitable cells expressing the receptor, the production of cAMP or any other suitable parameter can be used to monitor agonist activity on the receptor.

[0185] The skilled person will know suitable assay formats and examples are provided below. For example, the assay can utilize the human calcitonin receptor (hCT-R, e.g. isoform 2 of hCT-R) or the hAMYR3 receptor (see examples below). Where reference is made to the sequence of a precursor protein, it will be understood that the assay can utilize the mature protein lacking the signal sequence.

[0186] EC 50 The value can be used as a numerical measure of the agonist potency of a given receptor. The EC 50 value is a measure of the concentration of compound required to achieve half the maximum activity of the compound in a particular assay. Thus, for example, a compound having an EC 50 [hCT-R] lower than the EC 50 [hCT-R] of native human amylin, or lower than the EC 50 [hCT-R] of pramlintide. A compound having an EC

[0187] In some embodiments of the compounds of the application, the EC 50 is lower than about 1.5 nM (e.g. about 0.001 to about 1.5 nM).

[0188] In some embodiments of the compounds of the application, the EC 50 is lower than about 0.9 nM (e.g. about 0.001 to about 0.9 nM).

[0189] In some embodiments of the compounds of the application, the EC 50 is lower than about 0.5 nM (e.g. about 0.001 to about 0.5 nM).

[0190] In some embodiments of the compounds of the application, the EC 50 is lower than about 0.3 nM (e.g. about 0.001 to about 0.3 nM).

[0191] In some embodiments of the compounds of the application, the EC 50 is lower than about 0.2 nM (e.g. about 0.001 to about 0.2 nM).

[0192] The EC 50 of a compound against the hCT-R can be an indication of the effect of the compound on food intake, weight gain and / or weight loss. A compound having a lower EC 50 value against the hCT-R can have a greater effect on these parameters.

[0193] In some embodiments of the compounds of the application, the EC50 for hAMYR3 50 less than about 1.0 nM (e.g., about 0.001 to about 1.0 nM).

[0194] In some embodiments of the compounds of the application, the EC50 for hAMYR3 50 less than about 0.5 nM (e.g., about 0.001 to about 0.5 nM).

[0195] In some embodiments of the compounds of the application, the EC50 for hAMYR3 50 less than about 0.4 nM (e.g., about 0.001 to about 0.4 nM).

[0196] In some embodiments of the compounds of the application, the EC50 for hAMYR3 50 less than about 0.3 nM (e.g., about 0.001 to about 0.3 nM).

[0197] In some embodiments of the compounds of the application, the EC50 for hAMYR3 50 less than about 0.2 nM (e.g., about 0.001 to about 0.2 nM).

[0198] the EC50 for hCT-R (when expressed in the absence of hRAMP1, hRAMP2, and hRAMP3) 50 may be less than the EC50 for any or all of hAMYR1, hAMYR2, and hAMYR3 (e.g., for hAMYR3) 50 .

[0199] For example, the EC50 for hCT-R (when expressed in the absence of hRAMP1, hRAMP2, and hRAMP3) 50 may be more than 10-fold lower than the EC50 for any or all of hAMYR1, hAMYR2, and hAMYR3 (e.g., for hAMYR3) 50 .

[0200] the EC50 for hCT-R (when expressed in the absence of hRAMP1, hRAMP2, and hRAMP3) 50 may be more than 5-fold lower than the EC50 for any or all of hAMYR1, hAMYR2, and hAMYR3 (e.g., for hAMYR3) 50 .

[0201] the EC50 for hCT-R (when expressed in the absence of hRAMP1, hRAMP2, and hRAMP3) 50It can be substantially equal to (e.g., + / - 50%) the EC for any or all of hAMYR1, hAMYR2, and hAMYR3 (e.g., for hAMYR3). 50 .

[0202] ECG of hCT-R (when expressed in the absence of hRAMP1, hRAMP2, and hRAMP3) 50 It can be higher than EC for any or all of hAMYR1, hAMYR2, and hAMYR3 (e.g., for hAMYR3). 50 .

[0203] gastric emptying

[0204] Gastric emptying refers to the process by which the contents of the stomach are emptied into the duodenum.

[0205] Alterations in gastric emptying can affect the residence time of a given molecule in the stomach, and thus alter the time it takes for that molecule to reach a given downstream compartment (e.g., the duodenum, small intestine, and large intestine). Therefore, gastric emptying can delay the absorption of orally administered drugs.

[0206] In addition, delayed gastric emptying can affect blood glucose levels and thus affect any accompanying medications used to control them (such as non-oral medications), such as insulin.

[0207] It is known that peptide hormones in the prior art (especially peptide hormones used for weight loss or treatment of obesity and diabetes) cause delayed gastric emptying.

[0208] For example, tirzepatide (marketed under the brand names Mountaro and Zepbound) delays gastric emptying (Urva et al., Diabetes Obes Metab 2020 22 1886-1891), and therefore may affect the absorption of concomitant oral medications. Caution is advised when oral medications are administered concurrently with Mountaro. When administered concurrently with Mountaro, physicians are advised to monitor subjects taking oral medications that are effective only at threshold concentrations and those with narrow therapeutic indices (e.g., warfarin). It is recommended that subjects using oral hormonal contraceptives switch to non-oral contraceptive methods, or add barrier contraceptive methods 4 weeks after initial use of Mountaro and 4 weeks after each dose escalation of Mountaro.

[0209] In particular, prior art amylin analogs are known to cause delayed gastric emptying. This delay has an effect on concomitant medications. For example, the recommended administration of the amylin analog pralintide (… The object in Apply at least 1 hour before or Certain concomitant oral medications are administered at least 2 hours after administration of the amylin analogue.

[0210] As a further example, subjects receiving the amylin analogue pramlintide (Symlin®) are also informed of the risk of severe hypoglycaemia and advised to reduce the insulin dose (including pre-mixed insulin doses) at meals by 50% to reduce the risk of hypoglycaemia and to monitor blood glucose closely thereafter.

[0211] Thus, in addition to dosing regimens which facilitate where the timing of the concomitant agent is not affected by the administration of the amylin analogue of the application, the present application also provides for regimens where the dose of the concomitant agent is not affected by the administration of the amylin analogue of the application.

[0212] Furthermore, if the subject has taken a medication which alters gastrointestinal motility, they are also advised not to administer .

[0213] The amylin analogue of the application does not affect gastric emptying. In other words, the amylin analogue of the application does not slow gastric emptying.

[0214] It is envisaged that such considerations would not apply to the amylin analogue of the application due to the absence of delayed gastric emptying.

[0215] Gastric emptying is measured by the paracetamol absorption test. In this test, a fasted subject is given an oral dose of paracetamol at the same time as a standard meal. The concentration of paracetamol in the blood is observed after the meal. This can be used as an indication of the relative speed of gastric emptying when the concentration of paracetamol in the blood of a subject treated with an amylin analogue is compared to the paracetamol concentration in a subject treated with placebo. Using the paracetamol test it is easy to determine whether a given amylin analogue affects (i.e. slows) gastric emptying.

[0216] Concomitant agent

[0217] In the dosing regimens, methods and medical uses of the application, the amylin analogue of the application is administered to the subject within about 120 minutes of the concomitant agent (e.g. at the same time as the concomitant agent).

[0218] A concomitant agent is a substance (e.g. a medication, compound, molecule or supplement) which is used (e.g. administered to a subject) at the same time or at a similar time (e.g. within two hours) as the amylin analogue. A "concomitant agent" can also be referred to as "additional substance". The term "concomitant agent" does not imply or suggest any relationship between the amylin analogue and the concomitant agent other than a temporal relationship (i.e. they are administered at similar times).

[0219] A concomitant agent is distinct from an amylin analogue which does not affect the rate of gastric emptying in a subject.​

[0220] The concomitant agent can be a drug (i.e. an agent that treats a disease or condition). The concomitant agent can be a non-drug, for example a nutritional supplement. The concomitant agent can be any substance that is not an amylin analogue. The term "concomitant agent" can refer to any number of concomitant agents that are not amylin analogues.

[0221] The concomitant agent can be any substance whose administration is affected by gastric emptying rate. Preferably, the concomitant agent is a substance whose effectiveness is affected by gastric emptying rate. For example, where the therapeutic efficacy requires the drug to act quickly or to reach a threshold concentration, the concomitant agent - particularly a drug that is administered orally - can be dependent on gastric emptying rate. Such drugs include analgesics, antibiotics and contraceptives.

[0222] In some embodiments, the concomitant agent is a drug that requires rapid onset of action. In some embodiments, the concomitant agent is an oral drug that requires rapid onset of action.

[0223] In some embodiments, the concomitant agent is an oral drug.

[0224] In some embodiments, the concomitant agent is an analgesic.

[0225] In some embodiments, the concomitant agent is an antibiotic.

[0226] In some embodiments, the concomitant agent is an oral contraceptive.

[0227] In some embodiments, the concomitant agent is a nutritional supplement.

[0228] In some embodiments, the concomitant agent is a non-oral drug.

[0229] In some embodiments, the concomitant agent is administered intravenously, intramuscularly or subcutaneously.

[0230] In some embodiments, the concomitant agent is selected from the group consisting of: analgesics, antibiotics, anticoagulants, contraceptives, anti-hypotensive drugs, statins, antifungals, hypoglycaemic agents, contraceptives, oral drugs with narrow therapeutic index, and nutritional supplements.

[0231] In some embodiments, the concomitant agent is selected from the group consisting of: insulin, acetaminophen, digoxin, warfarin, metformin, hydrocortisone, griseofulvin, glibenclamide, gliclazide and glipizide.

[0232] In some embodiments, the concomitant agent is a sulfonylurea, for example glibenclamide, gliclazide, glipizide.

[0233] In some embodiments, the concomitant agent is insulin.

[0234] The subject can be administered one or more concomitant agents independently as defined herein. For example, where a amylin analogue of the application is administered to a subject simultaneously with, for example, three drugs, each of the three drugs will be considered independently to be a "concomitant agent" according to the application.

[0235] Thus, in some embodiments, the amylin analogue is administered to a subject simultaneously (within about 120 minutes of) at least one concomitant agent, for example at least two concomitant agents, at least three concomitant agents, at least four concomitant agents, at least five concomitant agents, at least six concomitant agents, at least seven concomitant agents, at least eight concomitant agents, at least nine concomitant agents, or at least ten concomitant agents.

[0236] In some embodiments, the amylin analogue is administered to a subject simultaneously (within about 120 minutes of) one or more concomitant agents, for example two or more concomitant agents, three or more concomitant agents, four or more concomitant agents, five or more concomitant agents, six or more concomitant agents, seven or more concomitant agents, eight or more concomitant agents, nine or more concomitant agents, or ten or more concomitant agents.

[0237] In some embodiments, the amylin analogue is administered to a subject simultaneously (within about 120 minutes of) one or more concomitant agents, for example two or more concomitant agents, three or more concomitant agents, four or more concomitant agents, five or more concomitant agents, six or more concomitant agents, seven or more concomitant agents, eight or more concomitant agents, nine or more concomitant agents, or ten or more concomitant agents.

[0238] Timing of administration

[0239] Surprisingly, the amylin analogues of the application do not cause any significant delay in gastric emptying. Thus, these compounds are suitable for administration simultaneously with, or in close time proximity to, other compounds or drugs (i.e. concomitant agents).

[0240] In particular, where the drug is to be administered orally, its administration can be sensitive to gastric emptying, and its administration will depend on the administration of any other drug that delays gastric emptying (e.g. an amylin analogue of the prior art).

[0241] For example, it can be necessary to delay administration of the drug until 2 hours or more has elapsed since administration of the amylin analogue that delays gastric emptying. Alternatively, it can be necessary to advance administration of the drug (or delay administration of the amylin analogue) so that the amylin analogue is administered at least one hour after administration of the drug has elapsed.

[0242] In both cases, the administration of the drug is time dependent on each other. Thus, the subject can have to change an existing dosing regimen, develop a burdensome new administration regimen, and / or face the risk of a drug-drug interaction that reduces the efficacy of the non-amylin analogue drug.

[0243] Advantageously, the amylin analogue of the present application has no effect on gastric emptying - i.e. does not delay gastric emptying like amylin and other hormone peptide treatments as well as amylin analogues - which means that the timing of the administration of one or more other (non-amylin analogue) substances to the subject does not need to be adjusted to take into account the administration of the amylin analogue of the present application. For example, unlike taspoglutide or pramlintide, the amylin analogue of the present application does not advantageously delay gastric emptying.

[0244] Thus, the timing of the administration of the amylin analogue does not affect the timing of the administration of the concomitant agent. Thus, the administration of the amylin analogue and the administration of the concomitant agent are substantially independent in time. In other words, the amylin analogue of the present application allows the independent administration in time of the amylin analogue of the present application and one or more concomitant agents.

[0245] Thus, in the dosing regimen, method and medical use of the present application, the amylin analogue of the present application is administered to the subject within about 120 minutes of the administration of the concomitant agent to the subject (e.g. simultaneously with the administration of the concomitant agent to the subject).

[0246] It is understood that the concomitant agent can be administered at any time (e.g. the time dictated by any dosing regimen of said concomitant agent) when not administered concomitantly with the amylin analogue of the present application. Thus, the administration of the concomitant agent is not limited by the administration of the amylin analogue.

[0247] The amylin analogue and the concomitant agent can be administered independently of each other. It is understood that the order in which the amylin analogue and the concomitant agent are presented herein does not limit the order in which they are administered. Thus, the amylin analogue can be administered before, simultaneously with, or after the concomitant agent, and vice versa.

[0248] The present application allows the administration of the amylin analogue to the subject while also administering one or more concomitant agents to the subject. Indeed, the amylin analogue and the concomitant agent can be administered to the subject in close proximity in time (i.e. at similar times), but not necessarily at the exact same moment. The key factor is that the present application allows the administration of the concomitant agent at a time point relative to the administration of the amylin analogue that would not be possible if it was not for the amylin analogue of the present application not delaying gastric emptying.

[0249] Accordingly, in some embodiments, the amylin analogue is administered to the subject within about 120 minutes of the administration of the concomitant agent to the subject, e.g., within about 110 minutes, e.g., within about 100 minutes, e.g., within about 90 minutes, e.g., within about 80 minutes, e.g., within about 70 minutes, e.g., within about 60 minutes, e.g., within about 50 minutes, e.g., within about 45 minutes, e.g., within about 40 minutes, e.g., within about 30 minutes, e.g., within about 20 minutes, e.g., within about 15 minutes, e.g., within about 10 minutes, e.g., within about 9 minutes, e.g., within about 8 minutes, e.g., within about 7 minutes, e.g., within about 6 minutes, e.g., within about 5 minutes, e.g., within about 4 minutes, e.g., within about 3 minutes, e.g., within about 2 minutes, or e.g., within about 1 minute of the administration of the concomitant agent to the subject.

[0250] In some embodiments, the amylin analogue is administered to the subject prior to the administration of the concomitant agent to the subject. In some embodiments, the amylin analogue is administered to the subject about 120 minutes prior to the administration of the concomitant agent to the subject, e.g., about 110 minutes prior, e.g., about 100 minutes prior, e.g., about 90 minutes prior, e.g., about 80 minutes prior, e.g., about 70 minutes prior, e.g., about 60 minutes prior, e.g., about 50 minutes prior, e.g., about 45 minutes prior, e.g., about 40 minutes prior, e.g., about 30 minutes prior, e.g., about 20 minutes prior, e.g., about 15 minutes prior, e.g., about 10 minutes prior, e.g., about 9 minutes prior, e.g., about 8 minutes prior, e.g., about 7 minutes prior, e.g., about 6 minutes prior, e.g., about 5 minutes prior, e.g., about 4 minutes prior, e.g., about 3 minutes prior, e.g., about 2 minutes prior, or e.g., about 1 minute prior to the administration of the concomitant agent to the subject.

[0251] In some embodiments, the amylin analogue is administered to the subject after the administration of the concomitant agent to the subject. In some embodiments, the amylin analogue is administered to the subject about 120 minutes after the administration of the concomitant agent to the subject, e.g., about 110 minutes after, e.g., about 100 minutes after, e.g., about 90 minutes after, e.g., about 80 minutes after, e.g., about 70 minutes after, e.g., about 60 minutes after, e.g., about 50 minutes after, e.g., about 45 minutes after, e.g., about 40 minutes after, e.g., about 30 minutes after, e.g., about 20 minutes after, e.g., about 15 minutes after, e.g., about 10 minutes after, e.g., about 9 minutes after, e.g., about 8 minutes after, e.g., about 7 minutes after, e.g., about 6 minutes after, e.g., about 5 minutes after, e.g., about 4 minutes after, e.g., about 3 minutes after, e.g., about 2 minutes after, or e.g., about 1 minute after the administration of the concomitant agent to the subject.

[0252] In some embodiments, the amylin analogue is administered to the subject concurrently with the concomitant agent. Concurrently is understood to mean that administration occurs substantially simultaneously, e.g. within 5 minutes of each other.

[0253] Dosage and dosage regimen

[0254] In the present context, the term "dosage" refers to the amount of a given compound (amylin analogue or concomitant agent) administered to a subject in each administration event. The term "dosage regimen" refers to both the amount and the frequency of administration of a given compound to a subject.

[0255] The dosage regimen of the amylin analogue can be selected independently of the dosage regimen of the concomitant agent.

[0256] A typical dosage regimen of the amylin analogue used in the present application can be from about 0.0001 to about 100 mg / kg body weight per day or every other day, e.g. from about 0.0005 to about 50 mg / kg body weight per day or every other day, e.g. from about 0.001 to about 10 mg / kg body weight per day or every other day, e.g. from about 0.01 to about 1 mg / kg body weight per day or every other day, administered in one or more doses, e.g. one to three doses. The precise dosage regimen employed will depend inter alia on the nature and severity of the disease or condition to be treated, the gender, age, weight, and general condition of the subject to be treated, possible other concurrent diseases or conditions for which treatment is being or will be given, and other factors known to medical practitioners skilled in the art.

[0257] A typical dosage regimen of the amylin analogue used in the present application can be from about 0.5 mg to about 10.0 mg per day or every other day, from about 0.6 mg to about 7.5 mg per day or every other day, preferably from about 1.2 mg to about 7.5 mg per day or every other day, preferably from about 1.2 to about 6.0 mg per day or every other day, preferably from about 2.4 to about 6.0 mg per day or every other day, preferably from about 2.4 to about 4.0 mg per day or every other day, preferably from about 2.4 to about 3.5 mg per day or every other day.

[0258] In some embodiments, the amylin analogue is administered to the subject at a dosage of from about 0.04 mg to about 10 mg, e.g. from about 0.04 mg to about 7 mg, from about 0.04 mg to about 6 mg, from about 0.04 mg to about 4.4 mg, from about 0.04 mg to about 2.4 mg, from about 0.6 mg to about 6 mg, from about 0.6 mg to about 4.4 mg, from about 0.6 mg to about 2.4 mg, from about 0.7 mg to about 6 mg, from about 0.7 mg to about 4.4 mg, or from about 0.7 mg to about 2.4 mg.

[0259] In some embodiments, the amylin analog is administered to the subject at a dose of about 0.6 mg to about 4.8 mg, about 0.6 mg to about 3.6 mg, about 0.6 mg to about 2.4 mg, or about 0.6 mg to about 1.2 mg.

[0260] In some embodiments, the amylin analog is administered to the subject at a dose of about 1.2 mg to about 4.8 mg, about 1.2 mg to about 3.6 mg, or about 1.2 mg to about 2.4 mg. In some embodiments, the amylin analog is administered to the subject at a dose of about 2.4 mg to about 4.8 mg or about 2.4 mg to about 3.6 mg. In some embodiments, the amylin analog is administered to the subject at a dose of about 3.6 mg to about 4.8 mg.

[0261] In some embodiments, the amylin analog is administered to the subject at a dose of up to about 10 mg, for example, up to about 9.5 mg, up to about 9 mg, up to about 8.5 mg, up to about 8 mg, up to about 7.5 mg, up to about 7 mg, up to about 6.5 mg, up to about 6 mg, up to about 5.5 mg, up to about 5 mg, up to about 4.8 mg, up to about 4.5 mg, up to about 4 mg, up to about 3.6 mg, up to about 3.5 mg, up to about 3 mg, up to about 2.5 mg, up to about 2.4 mg, up to about 2 mg, up to about 1.5 mg, up to about 1.2 mg, up to about 1 mg, up to about 0.6 mg, up to about 0.5 mg, or up to about 0.25 mg.

[0262] In some embodiments, the amylin analog is administered to the subject at a dose of about 10 mg, for example, about 9.5 mg, about 9 mg, about 8.5 mg, about 8 mg, about 7.5 mg, about 7 mg, about 6.5 mg, about 6 mg, about 5.5 mg, about 5 mg, about 4.8 mg, about 4.5 mg, about 4.4 mg, about 4 mg, about 3.6 mg, about 3.5 mg, about 3.4 mg, about 3 mg, about 2.5 mg, about 2.4 mg, about 2 mg, about 1.5 mg, about 1.4 mg, about 1.2 mg, about 1 mg, about 0.6 mg, about 0.5 mg, about 0.35 mg, about 0.25 mg, about 0.16 mg, about 0.08 mg, or about 0.04 mg.

[0263] In some embodiments, the amylin analogue according to the application is administered at regular intervals. In some embodiments, the amylin analogue is administered to the subject once a day, once every two days, once a week, once every two weeks, once every three weeks, or once every four weeks. In some embodiments, the amylin analogue is administered to the subject once a month.

[0264] In some embodiments, the dose of amylin analogue is not the same during each administration. In some embodiments, the dose of amylin analogue is increased for each successive administration. In some embodiments, the dose of amylin analogue is increased for each successive administration until a desired maximum dose is reached. Optionally, the desired maximum dose can be maintained for a given period of time (i.e. a dose plateau). In some embodiments, the dose of amylin analogue is the same during each administration.

[0265] Advantageously, the amylin analogue of the application has no effect on gastric emptying - i.e. does not delay gastric emptying like amylin and other amylin analogues - which means that the dose or dosage regimen of one or more other (non-amylin analogue) substances administered to the subject need not be adjusted to take into account administration of the amylin analogue of the application.

[0266] Accordingly, administration of the amylin analogue does not affect the dose or dosage regimen of the concomitant agent administered. Thus, the dose or dosage regimen of the amylin analogue and the concomitant agent are essentially independent. In other words, the amylin analogue of the application allows independent dose selection for administration of the amylin analogue of the application and the one or more concomitant agents. Accordingly, administration of the concomitant agent is not limited by administration of the amylin analogue.

[0267] Accordingly, in the dosing regimens, methods and medical uses of the application, the amylin analogue of the application and the concomitant agent are administered to the subject, wherein the dose of the concomitant agent administered is not altered relative to the dose of the concomitant agent administered in the absence of the amylin analogue.

[0268] Furthermore, in the dosing regimens, methods and medical uses of the application, the amylin analogue of the application is administered to the subject within about 120 minutes of (e.g. concurrently with) administration of the concomitant agent to the subject, and the dose of the concomitant agent administered is not altered relative to the dose administered in the absence of the amylin analogue.

[0269] The concomitant agent can be a substance for which the administered dose is dependent on the rate of gastric emptying, optionally wherein the dependency is due to blood glucose levels or rate of change thereof.

[0270] According to the foregoing, the concomitant agent can be an oral hypoglycemic agent, such as a sulfonylurea. In other embodiments, the oral hypoglycemic agent is glibenclamide, gliclazide and / or glipizide.

[0271] Furthermore, according to the foregoing, the concomitant agent is insulin.

[0272] It will be appreciated that the dose or dosage regimen of the concomitant agent can be adjusted independently of the timing and / or dose of the administration of the amylin analogue.

[0273] Dosing regimen

[0274] The present application provides a dosing regimen for a subject, wherein the dosing regimen comprises:

[0275] (a) administering to the subject an amylin analogue that does not affect gastric emptying rate; and

[0276] (b) administering to the subject a concomitant agent;

[0277] wherein:

[0278] the amylin analogue and the concomitant agent are administered to the subject within about 120 minutes of each other; and

[0279] the amylin analogue comprises or consists of a peptide of the formula:

[0280]

[0281] wherein:

[0282] R 1 is hydrogen, C 1-4 acyl, benzoyl or C 1-4 alkyl, or a half-life extending moiety M, wherein M is optionally linked to Z via a linker moiety L;

[0283] R 2 is OH or NHR 3 , wherein R 3 is hydrogen or C 1-3 alkyl; and

[0284] Z is an amino acid sequence of the formula I:

[0285]

[0286] wherein:

[0287] X1is selected from Arg, Lys and Glu;

[0288] X3is selected from Gly, Gin and Pro;

[0289] X4 is selected from Thr and Glu;

[0290] X5 is selected from Ala and Leu;

[0291] X6 is selected from Thr and Ser;

[0292] X10 is selected from Glu and Gin;

[0293] X14 is selected from Aad, His, Asp, Asn, and Arg;

[0294] X17 is selected from Gin, His, and Thr;

[0295] X19-X20 is selected from Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly, and Ala-Asn, or is absent;

[0296] X27 is selected from Leu and Pro;

[0297] X32 is selected from Val and Thr;

[0298] X35 is selected from Asn and Ser;

[0299] X37 is selected from Hyp and Pro; and

[0300] X2 and X7 are amino acid residues whose side chains together form a lactam bridge;

[0301] Gly(Me) is N-methylglycine [also known as sarcosine (Sar)]

[0302] Ile(Me) is N-methylisoleucine

[0303] Aad is 2-amino adipic acid, e.g., (2S)-2-amino adipic acid [also known as (2S)-2- amino hexanedioic acid], also known as homoglutamic acid.

[0304] In some embodiments, the present application provides a dosing regimen for a subject, wherein the dosing regimen comprises:

[0305] (a) administering to the subject an amylin analog that does not affect gastric emptying rate; and

[0306] (b) administering to the subject a concomitant agent;

[0307] wherein:

[0308] the amylin analog and the concomitant agent are administered to the subject within about 120 minutes of each other; and

[0309] The amylin analogue comprises or consists of a peptide of the formula:

[0310]

[0311] wherein:

[0312] Gly(Me): N-methylglycine [also known as sarcosine (Sar)]

[0313] Ile(Me): N-methylisoleucine

[0314] Aad: 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2- amino hexanedioic acid] also known as homoglutamic acid

[0315] Hyp: 4-hydroxyproline, for example (2S,4R)-4-hydroxyproline [also known as (4R)-4- hydroxy-L-proline]

[0316] [19CD]-isoGlu: the backbone nitrogen of the Arg residue (present at position X1 of the peptide sequence Z of the amylin analogue) is covalently linked via an amide linkage to the side chain carboxyl of the Glu moiety; the 19-carboxy-nonadecanoyl group is covalently linked via an amide linkage to the alpha amino group of the Glu linker (as described herein).

[0317] In one embodiment, the present application provides a dosing regimen for a subject, wherein the dosing regimen comprises:

[0318] (a) administering to the subject an amylin analogue that does not affect gastric emptying rate; and

[0319] (b) administering to the subject a concomitant agent;

[0320] wherein:

[0321] the amylin analogue and the concomitant agent are administered to the subject within about 120 minutes of each other;

[0322] The amylin analogue comprises or consists of a peptide of the formula:

[0323]

[0324] wherein:

[0325] R 1 is hydrogen, C 1-4 acyl, benzoyl or C 1-4 alkyl, or a half-life extending moiety M, wherein M is optionally linked to Z via a linker moiety L;

[0326] R 2 is OH or NHR 3 wherein R 3 is hydrogen or C 1-3 alkyl; and

[0327] Z is an amino acid sequence of Formula I:

[0328]

[0329] wherein:

[0330] X1 is selected from Arg, Lys, and Glu;

[0331] X3 is selected from Gly, Gin, and Pro;

[0332] X4 is selected from Thr and Glu;

[0333] X5 is selected from Ala and Leu;

[0334] X6 is selected from Thr and Ser;

[0335] X10 is selected from Glu and Gin;

[0336] X14 is selected from Aad, His, Asp, Asn, and Arg;

[0337] X17 is selected from Gin, His, and Thr;

[0338] X19-X20 is selected from Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly, and Ala-Asn, or is absent;

[0339] X27 is selected from Leu and Pro;

[0340] X32 is selected from Val and Thr;

[0341] X35 is selected from Asn and Ser;

[0342] X37 is selected from Hyp and Pro; and

[0343] X2 and X7 are amino acid residues whose side chains together form a lactam bridge;

[0344] Gly(Me) is N-methylglycine [also known as sarcosine (Sar)]

[0345] Ile(Me) is N-methylisoleucine

[0346] Aad is 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2-aminoadipic acid], also known as homoglutamic acid;

[0347] Furthermore, the dose of the concomitant is unchanged relative to the dose of the concomitant administered in the absence of amylin analogues.

[0348] Methods to suppress weight gain or lose weight

[0349] This invention provides a method for inhibiting weight gain and / or reducing weight in a subject, wherein the method includes:

[0350] (a) Administration of amylin analogues to a subject that do not affect the rate of gastric emptying; and

[0351] (b) Applying a comorbid agent to the object;

[0352] in:

[0353] Amylin analogues and concomitants are administered to the subject within approximately 120 minutes of each other; and

[0354] Amylin analogues comprise peptides or pharmaceutically acceptable salts and / or derivatives of the following formula, or consist of peptides or pharmaceutically acceptable salts and / or derivatives of the following formula:

[0355]

[0356] in:

[0357] R 1 It is hydrogen, C 1-4 Acyl, benzoyl or C 1-4 An alkyl group, or a half-life-extending portion M, wherein M is optionally connected to Z via a connector portion L;

[0358] R 2 Is it OH or NHR? 3 , where R 3 Is it hydrogen or C? 1-3 Alkyl; and

[0359] Z is the amino acid sequence of formula I:

[0360]

[0361] in:

[0362] X1 is selected from Arg, Lys, and Glu;

[0363] X3 is selected from Gly, Gln, and Pro;

[0364] X4 is selected from Thr and Glu;

[0365] X5 was chosen from Ala and Leu;

[0366] X6 is selected from Thr and Ser;

[0367] X10 is selected from the group consisting of Glu and Gin;

[0368] X14 is selected from the group consisting of Aad, His, Asp, Asn and Arg;

[0369] X17 is selected from the group consisting of Gin, His and Thr;

[0370] X19-X20 is selected from the group consisting of Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly and Ala-Asn, or is absent;

[0371] X27 is selected from the group consisting of Leu and Pro;

[0372] X32 is selected from the group consisting of Val and Thr;

[0373] X35 is selected from the group consisting of Asn and Ser;

[0374] X37 is selected from the group consisting of Hyp and Pro; and

[0375] X2 and X7 are amino acid residues whose side chains together form a lactam bridge;

[0376] Gly(Me) is N-methylglycine [also known as sarcosine (Sar)]

[0377] Ile(Me) is N-methylisoleucine

[0378] Aad is 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2-aminoadipic acid], also known as homoglutamic acid.

[0379] In some embodiments, the present application provides a method of inhibiting weight gain and / or reducing body weight in a subject, wherein the method comprises:

[0380] (a) administering to the subject an amylin analogue that does not affect gastric emptying rate; and

[0381] (b) administering to the subject a concomitant agent;

[0382] wherein:

[0383] the amylin analogue and the concomitant agent are administered to the subject within about 120 minutes of each other; and

[0384] the amylin analogue comprises or consists of a peptide of the formula:

[0385]

[0386] wherein:

[0387] Gly(Me): N-methylglycine [also known as sarcosine (Sar)]

[0388] Ile(Me): N-methylisoleucine

[0389] Aad: 2-amino adipic acid, e.g. (2S)-2-amino adipic acid [also known as (2S)-2- amino hexanedioic acid] [also known as homoglutamic acid]

[0390] Hyp: 4-hydroxyproline, e.g. (2S,4R)-4-hydroxyproline [also known as (4R)-4- hydroxy-L-proline]

[0391] [19CD]-isoGlu: the backbone nitrogen of the Arg residue (present at position X1 of the peptide sequence Z of the amylin analogue) is covalently linked via an amide linkage to the side chain carboxyl group of the Glu moiety; the 19-carboxy-nonadecanoyl group is covalently linked via an amide linkage to the alpha amino group of the Glu linker (as described herein).

[0392] The inhibition of glucagon secretion and food intake associated with amylin and amylin analogues can contribute to weight loss, or at least slow down (i.e. reduce or inhibit) weight gain. Thus, the methods of the application can be expressed as methods of inhibiting or reducing weight gain in a subject, methods of promoting weight loss, methods of reducing food intake and / or methods of weight loss.

[0393] Non-therapeutic use

[0394] The amylin analogues of the application can be used for non-therapeutic purposes. In this regard, it can be desirable for a subject to reduce food intake to lose weight (or inhibit weight gain), e.g. for cosmetic reasons, although the subject is not overweight, obese, diabetic or suffering from any other body weight related disease. Administration of the amylin analogue can achieve these desired effects (reduction of food intake, weight loss and / or inhibition of weight gain). In such embodiments, administration of the amylin analogue will not be therapeutic, as the amylin analogue is not treating a disease or condition.

[0395] Likewise, the concomitant agent need not be a drug, and does not necessarily have a therapeutic effect. For example, a concomitant agent that is a nutritional supplement will not have a therapeutic effect.

[0396] Thus, in some embodiments, the dosing regimen of the application is a non-therapeutic dosing regimen. Thus, the application provides a non-therapeutic dosing regimen for a subject, wherein the dosing regimen comprises:

[0397] (a) administering to the subject an amylin analogue of the application that does not affect gastric emptying rate; and

[0398] (b) administering to the subject an accompaniment;

[0399] wherein:

[0400] the amylin analogue and the accompaniment are administered to the subject within about 120 minutes of each other.

[0401] In some embodiments, the method of inhibiting weight gain and / or reducing weight in a subject is a non-therapeutic method. Accordingly, the present application provides a non-therapeutic method of inhibiting weight gain and / or reducing weight in a subject, wherein the method comprises:

[0402] (a) administering to the subject an amylin analogue of the application that does not affect gastric emptying rate; and

[0403] (b) administering to the subject an accompaniment;

[0404] wherein:

[0405] the amylin analogue and the accompaniment are administered to the subject within about 120 minutes of each other.

[0406] In some embodiments, the method of inhibiting weight gain and / or reducing weight in a subject is a cosmetic method.

[0407] When the amylin analogue is used for therapeutic and non-therapeutic purposes, the considerations outlined herein in relation to gastric emptying and accompaniments (i.e. second drugs) apply equally.

[0408] Such non-therapeutic methods are intended to reduce the weight of a subject who is not obese or morbidly obese.

[0409] In some embodiments of the non-therapeutic methods of the application, the subject has a healthy weight. In other words, in some embodiments, the subject is not overweight, not obese and not morbidly obese. In some embodiments, the subject has a BMI of 18.5 to 24.9 kg / m 2 corresponding to a healthy weight.

[0410] In some embodiments of the non-therapeutic methods of the application, the subject is overweight. In other words, in some embodiments, the subject does not have a healthy weight, but is not obese and not morbidly obese. In some embodiments, the subject has a BMI of 25.0 to 29.9 kg / m 2 corresponding to being overweight.

[0411] Therapeutic uses

[0412] From the unexpected finding that the amylin analogues described herein do not stimulate gastric emptying delay, they have utility in a number of therapeutic settings. The amylin analogues of the application can be used to treat or prevent a disease.

[0413] More specifically, the amylin analogs of the present application are useful, inter alia, for reducing food intake, promoting weight loss, and inhibiting or reducing weight gain. Thus, they can provide an attractive therapeutic option for, inter alia, obesity and metabolic diseases caused by, characterized by, or associated with excess body weight. Treatment can be achieved, for example, by controlling appetite, eating, food intake, caloric intake, and / or energy expenditure.

[0414] The term "treatment" (and other grammatical variations thereof) as used in the context of the present application refers to an approach for obtaining beneficial or desired clinical results. For purposes of the present application, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. "Treatment" can also mean prolonging survival as compared to expected survival in the absence of treatment. "Treatment" is an intervention performed with the intention of preventing the development or altering the pathology of a disorder. Thus, "treatment" refers to both therapeutic treatment and prophylactic or preventative measures. Where used in the context of prophylactic or preventative measures, the amylin analog or concomitant agent need not completely prevent the occurrence of a disease or disorder. Those in need of treatment include those already with the disorder as well as those in which the disorder is to be prevented from occurring. "Treatment" also means inhibition or reduction of an increase in a pathological condition or symptom (e.g., weight gain or hypoglycemia), as compared to the absence of treatment, and does not necessarily imply that the relevant disorder is completely stopped or cured. The term "prevention" (and other grammatical variations thereof) refers to the prevention of a disease, for example, by alleviating symptoms of the disease.

[0415] The subject can take any number of other drugs (i.e., other than the amylin analogs described herein) to prevent or treat any number of the diseases or disorders described herein, or virtually any other disease or disorder. Thus, the amylin analogs described herein are particularly advantageous in that they do not interfere with the administration of such other drugs, as described elsewhere herein.

[0416] Method of treatment

[0417] Thus, the present application provides a method of treating or preventing a disease in a subject, wherein the method comprises:

[0418] (a) administering to the subject an amylin analog that does not affect gastric emptying rate; and

[0419] (b) administering to the subject a concomitant agent;

[0420] wherein:

[0421] the amylin analogue and the concomitant agent are administered to the subject within about 120 minutes of each other; and

[0422] The amylin analogue comprises or consists of a peptide of the formula:

[0423]

[0424] wherein:

[0425] R 1 is hydrogen, C 1-4 acyl, benzoyl or C 1-4 alkyl, or a half-life extending moiety M, wherein M is optionally linked to Z via a linker moiety L;

[0426] R 2 is OH or NHR 3 , wherein R 3 is hydrogen or C 1-3 alkyl; and

[0427] Z is an amino acid sequence of the formula I:

[0428]

[0429] wherein:

[0430] X1is selected from Arg, Lys and Glu;

[0431] X3is selected from Gly, Gin and Pro;

[0432] X4is selected from Thr and Glu;

[0433] X5is selected from Ala and Leu;

[0434] X6is selected from Thr and Ser;

[0435] X10is selected from Glu and Gin;

[0436] X14is selected from Aad, His, Asp, Asn and Arg;

[0437] X17is selected from Gin, His and Thr;

[0438] X19-X20is selected from Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly and Ala-Asn, or is absent;

[0439] X27is selected from Leu and Pro;

[0440] X32is selected from Val and Thr;

[0441] X35 is selected from Asn and Ser;

[0442] X37 is selected from Hyp and Pro; and

[0443] X2 and X7 are amino acid residues whose side chains together form a lactam bridge;

[0444] Gly(Me) is N-methylglycine [also known as sarcosine (Sar)]

[0445] Ile(Me) is N-methylisoleucine

[0446] Aad is 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2- amino hexanedioic acid] [also known as homoglutamic acid]

[0447] In some embodiments, the present application provides a method of treating or preventing a disease in a subject, wherein the method comprises:

[0448] (a) administering to the subject an amylin analogue that does not affect gastric emptying rate; and

[0449] (b) administering to the subject a concomitant agent;

[0450] wherein:

[0451] the amylin analogue and the concomitant agent are administered to the subject within about 120 minutes of each other; and

[0452] the amylin analogue comprises or consists of a peptide of the formula:

[0453]

[0454] wherein:

[0455] Gly(Me): N-methylglycine [also known as sarcosine (Sar)]

[0456] Ile(Me): N-methylisoleucine

[0457] Aad: 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2- amino hexanedioic acid] [also known as homoglutamic acid]

[0458] Hyp: 4-hydroxyproline, for example (2S,4R)-4-hydroxyproline [also known as (4R)-4- hydroxy-L-proline]

[0459] [19CD]-isoGlu: the backbone nitrogen of the Arg residue (present at position X1 of the peptide sequence Z of the amylin analogue) is covalently linked via an amide linkage to the side chain carboxyl group of the Glu moiety; the 19-carboxy-nonadecanoyl group is covalently linked via an amide linkage to the alpha amino group of the Glu linker (as described herein).

[0460] In some embodiments, the present application provides a method of treating a subject without altering (e.g. delaying) gastric emptying, the method comprising administering to a subject in need thereof an amylin analogue according to the present application.

[0461] In some embodiments, the present application provides a method for maintaining a normal administration regimen of one or more concomitantly administered drugs in a subject in need of treatment with an amylin analogue, wherein the method comprises administering to the subject an amylin analogue according to the present application.

[0462] Medical use

[0463] The present application also provides an amylin analogue that does not affect the rate of gastric emptying for use in a method of treating or preventing a disease in a subject, wherein the method comprises:

[0464] (a) administering to the subject an amylin analogue; and

[0465] (b) administering to the subject a concomitant agent;

[0466] wherein:

[0467] the amylin analogue and the concomitant agent are administered to the subject within about 120 minutes of each other; and

[0468] the amylin analogue comprises or consists of a peptide of the formula:

[0469]

[0470] wherein:

[0471] R 1 is hydrogen, C 1-4 acyl, benzoyl or C 1-4 alkyl, or a half-life extending moiety M, wherein M is optionally linked to Z via a linker moiety L;

[0472] R 2 is OH or NHR 3 , wherein R 3 is hydrogen or C 1-3 alkyl; and

[0473] Z is an amino acid sequence of the formula I:

[0474]

[0475] wherein:

[0476] X1 is selected from Arg, Lys and Glu;

[0477] X3 is selected from Gly, Gin and Pro;

[0478] X4 is selected from Thr and Glu;

[0479] X5 is selected from Ala and Leu;

[0480] X6 is selected from Thr and Ser;

[0481] X10 is selected from Glu and Gin;

[0482] X14 is selected from Aad, His, Asp, Asn and Arg;

[0483] X17 is selected from Gin, His and Thr;

[0484] X19-X20 is selected from Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly and Ala-Asn, or is absent;

[0485] X27 is selected from Leu and Pro;

[0486] X32 is selected from Val and Thr;

[0487] X35 is selected from Asn and Ser;

[0488] X37 is selected from Hyp and Pro; and

[0489] X2 and X7 are amino acid residues whose side chains together form a lactam bridge;

[0490] Gly(Me) is N-methylglycine [also known as sarcosine (Sar)]

[0491] Ile(Me) is N-methylisoleucine

[0492] Aad is 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2- amino hexanedioic acid], also known as homoglutamic acid.

[0493] The present application also provides the use of an amylin analogue which does not affect gastric emptying rate for the manufacture of a medicament for use in a method of treating or preventing a disease in a subject, wherein the method comprises:

[0494] (a) administering an amylin analogue to the subject; and

[0495] (b) administering to the subject an accompaniment agent;

[0496] wherein:

[0497] the amylin analogue and the accompaniment agent are administered to the subject within about 120 minutes of each other; and

[0498] the amylin analogue comprises or consists of a peptide of the formula:

[0499]

[0500] wherein:

[0501] R 1 is hydrogen, C 1-4 acyl, benzoyl or C 1-4 alkyl, or a half-life extending moiety M, wherein M is optionally linked to Z via a linker moiety L;

[0502] R 2 is OH or NHR 3 , wherein R 3 is hydrogen or C 1-3 alkyl; and

[0503] Z is an amino acid sequence of the formula I:

[0504]

[0505] wherein:

[0506] X1is selected from Arg, Lys and Glu;

[0507] X3is selected from Gly, Gin and Pro;

[0508] X4is selected from Thr and Glu;

[0509] X5is selected from Ala and Leu;

[0510] X6is selected from Thr and Ser;

[0511] X10is selected from Glu and Gin;

[0512] X14is selected from Aad, His, Asp, Asn and Arg;

[0513] X17is selected from Gin, His and Thr;

[0514] X19-X20 is selected from Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly and Ala-Asn, or is absent;

[0515] X27 is selected from Leu and Pro;

[0516] X32 is selected from Val and Thr;

[0517] X35 is selected from Asn and Ser;

[0518] X37 is selected from Hyp and Pro; and

[0519] X2 and X7 are amino acid residues whose side chains together form a lactam bridge;

[0520] Gly(Me) is N-methylglycine [also known as sarcosine (Sar)]

[0521] Ile(Me) is N-methylisoleucine

[0522] Aad is 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2- amino hexanedioic acid], also known as homoglutamic acid.

[0523] In some embodiments, the present application provides an amylin analogue according to the present application for use in therapy, wherein the amylin analogue does not alter gastric emptying.

[0524] In some embodiments, the present application provides an amylin analogue according to the present application for use in therapy, wherein the amylin analogue does not slow gastric emptying.

[0525] In some embodiments, the present application provides an amylin analogue according to the present application for use in a method of treating or preventing obesity, overweight and / or diabetes while maintaining normal gastric emptying.

[0526] In some embodiments, the present application provides an amylin analogue according to the present application for use in a method of treating or preventing obesity, overweight and / or diabetes without delaying gastric emptying.

[0527] The body weight of a subject can be referred to using the Body Mass Index (BMI), which for a human subject is calculated by dividing the body weight of the subject in kilograms by the square of the height of the subject in metres.

[0528] In some embodiments, the subject is obese. In some embodiments, the subject has a BMI of 30.0 to 39.9 kg / m2. 2 corresponding to obesity.

[0529] In some embodiments, the subject is morbidly obese. In some embodiments, the subject has a BMI of 40.0 kg / m 2 or higher, corresponding to morbid obesity.

[0530] In some embodiments, the present application provides a method for treating a subject in need thereof with an amylin analogue according to the present application, wherein the subject is receiving one or more concomitant medications.

[0531] Disease

[0532] The terms "disease", "disorder" and "condition" are synonymous and used interchangeably herein to refer to a state of dysfunction of the body.

[0533] The amylin analogues of the present application can be used to treat obesity and related diseases, conditions and health states, including but not limited to morbid obesity, pre-surgical obesity, obesity-related inflammation, obesity-related gallbladder disease, and obesity-induced sleep apnea and respiratory problems, cartilage degradation, osteoarthritis, and reproductive health complications of obesity or overweight (e.g. infertility).

[0534] Accordingly, in some embodiments of the dosing regimen, method and medical use of the present application, the disease is overweight, obesity, morbid obesity, diabetes, or a disease associated with obesity or diabetes. In other words, in some embodiments of the dosing regimen, method and medical use of the present application, the subject is overweight, has obesity, has morbid obesity, has diabetes, or has a disease associated with obesity or diabetes. The diabetes can be type 1 diabetes or type 2 diabetes.

[0535] In some embodiments of the dosage regimen, methods and medical uses of the application, the disease associated with obesity or diabetes is selected from the group consisting of obesity-associated inflammation, obesity-associated gallbladder disease, obesity-induced sleep apnea, obesity-associated respiratory problems, cartilage degeneration, osteoarthritis, infertility, Alzheimer's disease, prediabetes, insulin resistance syndrome, impaired glucose tolerance (IGT), a disease state associated with elevated blood glucose levels, metabolic disease, metabolic syndrome, hyperglycemia, hypertension, atherogenic dyslipidemia, hepatic steatosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), renal failure, arteriosclerosis, macrovascular disease, microvascular disease, diabetic heart disease, diabetic cardiomyopathy, heart failure as a complication of diabetes, coronary heart disease, peripheral arterial disease, and stroke.

[0536] The compounds can also be used to lower circulating LDL levels and / or to increase the HDL / LDL ratio.

[0537] The effects of the above-mentioned compounds can be mediated in whole or in part by an effect on body weight, or can be independent thereof.

[0538] Metabolic syndrome is characterized by a cluster of metabolic risk factors in one person. They include abdominal obesity (excess fat tissue around the internal organs in the abdomen), atherogenic dyslipidemia (dyslipidemia, including high triglycerides, low HDL cholesterol and / or high LDL cholesterol, which promotes plaque formation in the walls of arteries), elevated blood pressure (hypertension), insulin resistance and glucose intolerance, prothrombotic state (e.g. high fibrinogen or plasminogen activator inhibitor-1 in the blood) and proinflammatory state (e.g. elevated C-reactive protein in the blood).

[0539] Individuals with metabolic syndrome are at increased risk for coronary heart disease and other diseases related to manifestations of arteriosclerosis, such as stroke and peripheral vascular disease. The main underlying risk factor for this syndrome appears to be abdominal obesity.

[0540] Pharmaceutical compositions

[0541] Pharmaceutical compositions comprising an amylin analogue are described herein. As with all aspects of the application, it is to be understood that reference to an amylin analogue encompasses reference to pharmaceutically acceptable salts and solvates.

[0542] The amylin analogs of the present application can be formulated into pharmaceutical compositions suitable for administration with or without storage, and typically comprise a therapeutically effective amount of at least one peptide of the present application, together with a pharmaceutically acceptable carrier, excipient, or vehicle.

[0543] The term "pharmaceutically acceptable carrier" includes any of the standard pharmaceutical carriers. Pharmaceutically acceptable carriers for therapeutic use are well known in the pharmaceutical art, and are described, for example, in "Remington's Pharmaceutical Sciences", 17th Ed., Alfonso R. Gennaro (ed.), Mark Publishing Company, Easton, PA, USA, 1985. For example, sterile saline and phosphate-buffered saline at mild or physiological pH can be used. Suitable pH buffers can be, for example, phosphate, citrate, acetate, tris(hydroxymethyl)-aminomethane (TRIS), N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPS), ammonium bicarbonate, diethanolamine, histidine, arginine, lysine or acetate (e.g. as sodium acetate), or mixtures thereof. The term also encompasses any carrier substance listed in the US Pharmacopeia for use in animals, including humans.

[0544] The pharmaceutical compositions of the present application can be in unit dosage form. In such form, the composition is divided into unit doses containing appropriate quantities of one or more of the active components. The unit dosage form can be present as a package of discrete quantities such as packaged tablets, capsules or a bulk container of powder or granules. The unit dosage form can also be, for example, a capsule, cachet or tablet itself, or it can be an appropriate number of any of these packaged forms. The unit dosage form can also be provided in a single dose injectable form, for example in the form of a pen device containing a liquid (typically aqueous) composition. The composition can be formulated for any suitable route and mode of administration. Pharmaceutically acceptable carriers or diluents include those used in formulations suitable for, for example, oral, intravitreal, rectal, vaginal, nasal, topical, enteral or parenteral (including subcutaneous (sc), intramuscular (im), intravenous (iv), intradermal and transdermal) administration or administration by inhalation. The formulation can conveniently be presented in unit dosage form and can be prepared by any methods well known in the art of pharmacy.

[0545] In some cases, subcutaneous or transdermal modes of administration can be suitable for the peptides of the present application.

[0546] Other embodiments relate to devices, dosage forms, and packages for delivering the pharmaceutical formulations of the application. Thus, at least one peptide in a stable or preserved formulation or solution described herein can be administered to a patient according to the application by a variety of delivery methods, including by sc or im injection, or by transdermal, pulmonary, or transmucosal administration, or by implant, or by use of an osmotic pump, cartridge, micro-pump, or other means recognized by those skilled in the art.

[0547] Other embodiments relate to oral formulations and oral administration. Formulations for oral administration can rely on the co-administration of a vehicle (e.g., resorcinol and / or a non-ionic surfactant such as polyoxyethylene oleyl ether and n-hexadecyl polyethylene ether) to artificially increase the permeability of the intestinal wall, and / or the co-administration of an enzyme inhibitor (e.g., pancreatic trypsin inhibitor, diisopropylfluorophosphate (DFF), or trasylol) to inhibit enzymatic degradation. Active ingredient compounds in solid dosage forms for oral administration can be mixed with at least one additive such as sucrose, lactose, cellulose, mannitol, trehalose, raffinose, maltitol, dextran, starch, agar, alginate, chitin, chitosan, pectin, tragacanth gum, gum arabic, gelatin, collagen, casein, albumin, synthetic or semi-synthetic polymers, or glycerides. These dosage forms can also contain other types of additives such as inactive diluents, lubricants (e.g., magnesium stearate), parabens, preservatives (e.g., sorbic acid, ascorbic acid, or alpha-tocopherol), antioxidants (e.g., cysteine), disintegrants, binding agents, thickening agents, buffers, pH adjusting agents, sweeteners, flavoring agents, or perfuming agents.

[0548] Apparatus and kits

[0549] In some embodiments, the application relates to a device comprising an amylin analogue or a pharmaceutical composition of the application for use in delivering the analogue to a subject. Through such a device, the amylin analogue can be administered to the subject by a variety of delivery methods, including: intravenous, subcutaneous, intramuscular, or intraperitoneal injection; oral administration; transdermal administration; pulmonary or transmucosal administration; administration by implant, osmotic pump, cartridge, or micro-pump; or by other means recognized by those skilled in the art.

[0550] In some embodiments, the application relates to a kit comprising an amylin analogue of the application or a pharmaceutical composition thereof and one or more second medicaments. In certain embodiments, the kit further comprises packaging and / or instructions for use.

[0551] Aspects of the invention

[0552] The invention is described by the following numbered aspects.

[0553] Aspect 1. A dosing regimen for a subject, wherein the dosing regimen comprises:

[0554] (a) administering to the subject an amylin analogue that does not affect gastric emptying rate; and

[0555] (b) administering to the subject a companion agent;

[0556] wherein:

[0557] the amylin analogue and the companion agent are administered to the subject within about 120 minutes of each other; and

[0558] the amylin analogue comprises or consists of a peptide of the formula:

[0559]

[0560] wherein:

[0561] R 1 is hydrogen, C 1-4 acyl, benzoyl or C 1-4 alkyl, or a half-life extending moiety M, wherein M is optionally linked to Z via a linker moiety L;

[0562] R 2 is OH or NHR 3 , wherein R 3 is hydrogen or C 1-3 alkyl; and

[0563] Z is an amino acid sequence of Formula I:

[0564]

[0565] wherein:

[0566] X1is selected from Arg, Lys and Glu;

[0567] X3is selected from Gly, Gin and Pro;

[0568] X4is selected from Thr and Glu;

[0569] X5is selected from Ala and Leu;

[0570] X6is selected from Thr and Ser;

[0571] X10is selected from Glu and Gin;

[0572] X14 is selected from the group consisting of Aad, His, Asp, Asn and Arg;

[0573] X17 is selected from the group consisting of Gin, His and Thr;

[0574] X19-X20 is selected from the group consisting of Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly and Ala-Asn, or is absent;

[0575] X27 is selected from the group consisting of Leu and Pro;

[0576] X32 is selected from the group consisting of Val and Thr;

[0577] X35 is selected from the group consisting of Asn and Ser;

[0578] X37 is selected from the group consisting of Hyp and Pro; and

[0579] X2 and X7 are amino acid residues whose side chains together form a lactam bridge;

[0580] Gly(Me) is N-methylglycine [also known as sarcosine (Sar)]

[0581] Ile(Me) is N-methylisoleucine

[0582] Aad is 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2- amino hexanedioic acid], also known as homoglutamic acid.

[0583] Aspect 2. The dosing regimen of aspect 1, wherein the dosing regimen is a non- therapeutic dosing regimen.

[0584] Aspect 3. The dosing regimen of aspect 1 or aspect 2, wherein the subject:

[0585] (i) is overweight;

[0586] (ii) has obesity;

[0587] (iii) has morbid obesity;

[0588] (iv) has diabetes; and / or

[0589] (v) has a disease associated with obesity or diabetes.

[0590] Aspect 4. A method of inhibiting weight gain and / or reducing body weight in a subject, wherein the method comprises:

[0591] (a) administering to the subject an amylin analogue that does not affect gastric emptying rate; and

[0592] (b) administering to the subject a companion agent;

[0593] wherein:

[0594] the amylin analogue and the companion agent are administered to the subject within about 120 minutes of each other; and

[0595] the amylin analogue comprises or consists of a peptide of the formula:

[0596]

[0597] wherein:

[0598] R 1 is hydrogen, C 1-4 acyl, benzoyl or C 1-4 alkyl, or a half-life extending moiety M, wherein M is optionally linked to Z via a linker moiety L;

[0599] R 2 is OH or NHR 3 , wherein R 3 is hydrogen or C 1-3 alkyl; and

[0600] Z is an amino acid sequence of the formula I:

[0601]

[0602] wherein:

[0603] X1is selected from Arg, Lys and Glu;

[0604] X3is selected from Gly, Gin and Pro;

[0605] X4is selected from Thr and Glu;

[0606] X5is selected from Ala and Leu;

[0607] X6is selected from Thr and Ser;

[0608] X10is selected from Glu and Gin;

[0609] X14is selected from Aad, His, Asp, Asn and Arg;

[0610] X17is selected from Gin, His and Thr;

[0611] X19-X20is selected from Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly and Ala-Asn, or is absent;

[0612] X27 is selected from Leu and Pro;

[0613] X32 is selected from Val and Thr;

[0614] X35 is selected from Asn and Ser;

[0615] X37 is selected from Hyp and Pro; and

[0616] X2 and X7 are amino acid residues whose side chains together form a lactam bridge;

[0617] Gly(Me) is N-methylglycine [also known as sarcosine (Sar)]

[0618] Ile(Me) is N-methylisoleucine

[0619] Aad is 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2- amino hexanedioic acid], also known as homoglutamic acid.

[0620] Aspect 5. The method of aspect 4, wherein the method is a non-therapeutic method.

[0621] Aspect 6. The method of aspect 4 or aspect 5, wherein the subject:

[0622] (i) is overweight;

[0623] (ii) has obesity;

[0624] (iii) has morbid obesity;

[0625] (iv) has diabetes; and / or

[0626] (v) has a disease associated with obesity or diabetes.

[0627] Aspect 7. A method of treating or preventing a disease in a subject, wherein the method comprises:

[0628] (a) administering to the subject an amylin analogue that does not affect gastric emptying rate; and

[0629] (b) administering to the subject a companion agent;

[0630] wherein:

[0631] the amylin analogue and the companion agent are administered to the subject within about 120 minutes of each other; and

[0632] the amylin analogue comprises or consists of a peptide of the following formula:

[0633]

[0634] wherein:

[0635] R 1 is hydrogen, C 1-4 acyl, benzoyl or C 1-4 alkyl, or a half-life extending moiety M, wherein M is optionally linked to Z via a linker moiety L;

[0636] R 2 is OH or NHR 3 , wherein R 3 is hydrogen or C 1-3 alkyl; and

[0637] Z is an amino acid sequence of Formula I:

[0638]

[0639] wherein:

[0640] X1is selected from Arg, Lys and Glu;

[0641] X3is selected from Gly, Gin and Pro;

[0642] X4is selected from Thr and Glu;

[0643] X5is selected from Ala and Leu;

[0644] X6is selected from Thr and Ser;

[0645] X10is selected from Glu and Gin;

[0646] X14is selected from Aad, His, Asp, Asn and Arg;

[0647] X17is selected from Gin, His and Thr;

[0648] X19-X20is selected from Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly and Ala-Asn, or is absent;

[0649] X27is selected from Leu and Pro;

[0650] X32is selected from Val and Thr;

[0651] X35is selected from Asn and Ser;

[0652] X37is selected from Hyp and Pro; and

[0653] X2and X7are amino acid residues whose side chains together form a lactam bridge;

[0654] Gly(Me) is N-methylglycine [also known as sarcosine (Sar)]

[0655] Ile(Me) is N-methylisoleucine

[0656] Aad is 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2- amino hexanedioic acid], also known as homoglutamic acid.

[0657] Aspect 8. An amylin analogue which does not affect gastric emptying rate, for use in a method of treating or preventing a disease in a subject, wherein the method comprises:

[0658] (a) administering an amylin analogue to the subject; and

[0659] (b) administering a concomitant agent to the subject;

[0660] wherein:

[0661] the amylin analogue and the concomitant agent are administered to the subject within about 120 minutes of each other; and

[0662] the amylin analogue comprises or consists of a peptide of the formula:

[0663]

[0664] wherein:

[0665] R 1 is hydrogen, C 1-4 acyl, benzoyl or C 1-4 alkyl, or a half-life extending moiety M, wherein M is optionally linked to Z via a linker moiety L;

[0666] R 2 is OH or NHR 3 , wherein R 3 is hydrogen or C 1-3 alkyl; and

[0667] Z is an amino acid sequence of the formula I:

[0668]

[0669] wherein:

[0670] X1is selected from Arg, Lys and Glu;

[0671] X3is selected from Gly, Gin and Pro;

[0672] X4is selected from Thr and Glu;

[0673] X5 is selected from Ala and Leu;

[0674] X6 is selected from Thr and Ser;

[0675] X10 is selected from Glu and Gin;

[0676] X14 is selected from Aad, His, Asp, Asn and Arg;

[0677] X17 is selected from Gin, His and Thr;

[0678] X19-X20 is selected from Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly and Ala-Asn, or is absent;

[0679] X27 is selected from Leu and Pro;

[0680] X32 is selected from Val and Thr;

[0681] X35 is selected from Asn and Ser;

[0682] X37 is selected from Hyp and Pro; and

[0683] X2 and X7 are amino acid residues whose side chains together form a lactam bridge;

[0684] Gly(Me) is N-methylglycine [also known as sarcosine (Sar)]

[0685] Ile(Me) is N-methylisoleucine

[0686] Aad is 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2- amino hexanedioic acid], also known as homoglutamic acid.

[0687] Aspect 9. Use of an amylin analogue which does not affect gastric emptying rate for the manufacture of a medicament for use in a method of treating or preventing a disease in a subject, wherein the method comprises:

[0688] (a) administering an amylin analogue to the subject; and

[0689] (b) administering a concomitant agent to the subject;

[0690] wherein:

[0691] the amylin analogue and the concomitant agent are administered to the subject within about 120 minutes of each other; and

[0692] The amylin analogue comprises or consists of a peptide of the formula:

[0693]

[0694] wherein:

[0695] R 1 is hydrogen, C 1-4 1-4C-alkyl, acyl, benzoyl or C 1-4 1-4C-alkyl, or a half-life extending moiety M, wherein M is optionally linked to Z via a linker moiety L;

[0696] R 2 is OH or NHR 3 , wherein R 3 is hydrogen or C 1-3 1-4C-alkyl; and

[0697] Z is an amino acid sequence of the formula I:

[0698]

[0699] wherein:

[0700] X1is selected from Arg, Lys and Glu;

[0701] X3is selected from Gly, Gin and Pro;

[0702] X4is selected from Thr and Glu;

[0703] X5is selected from Ala and Leu;

[0704] X6is selected from Thr and Ser;

[0705] X10is selected from Glu and Gin;

[0706] X14is selected from Aad, His, Asp, Asn and Arg;

[0707] X17is selected from Gin, His and Thr;

[0708] X19-X20is selected from Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly and Ala-Asn, or is absent;

[0709] X27is selected from Leu and Pro;

[0710] X32is selected from Val and Thr;

[0711] X35is selected from Asn and Ser;

[0712] X37 is selected from Hyp and Pro; and

[0713] X2 and X7 are amino acid residues whose side chains together form a lactam bridge;

[0714] Gly(Me) is N-methylglycine [also known as sarcosine (Sar)]

[0715] Ile(Me) is N-methylisoleucine

[0716] Aad is 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2-amino hexanedioic acid], also known as homoglutamic acid.

[0717] Aspect 10. The method of aspect 7, the amylin analogue for use of aspect 8, or the use of aspect 9, wherein the disease is overweight, obesity, morbid obesity, diabetes, or a disease associated with obesity or diabetes.

[0718] Aspect 11. The dosage regimen of aspect 3, the method of aspect 6, or the method, the amylin analogue for use, or the use of aspect 10, wherein the diabetes is type 1 diabetes or type 2 diabetes.

[0719] Aspect 12. The dosage regimen of aspect 3, the method of aspect 6, or the method, the amylin analogue for use, or the use of aspect 10, wherein the disease associated with obesity or diabetes is selected from the group consisting of obesity-associated inflammation, obesity-associated gallbladder disease, obesity-induced sleep apnea, obesity-associated respiratory problems, cartilage degeneration, osteoarthritis, infertility, Alzheimer’s disease, prediabetes, insulin resistance syndrome, impaired glucose tolerance (IGT), a disease state associated with elevated blood glucose levels, metabolic disease, metabolic syndrome, hyperglycemia, hypertension, atherogenic dyslipidemia, hepatic steatosis, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), renal failure, arteriosclerosis, macrovascular disease, microvascular disease, diabetic heart disease, diabetic cardiomyopathy, heart failure as a complication of diabetes, coronary heart disease, peripheral arterial disease, and stroke.

[0720] Aspect 13. The dosage regimen, the method, the amylin analogue for use, or the use of any one of aspects 1 to 12, wherein the amylin analogue comprises or consists of a peptide of the following formula:

[0721]

[0722] and wherein the parentheses ( ) indicate an intramolecular lactam bridge formed between the side chains of the residues at sequence positions X2 and X7.

[0723] Aspect 14. The dosing regimen, the method, the amylin analogue for use, or the use of any one of aspects 1 to 13, wherein the amylin analogue and the concomitant agent are administered to the subject within 90 minutes of each other, within 60 minutes of each other, within 45 minutes of each other, within 30 minutes of each other, within 15 minutes of each other, and / or substantially simultaneously with each other.

[0724] Aspect 15. The dosing regimen, the method, the amylin analogue for use, or the use of any one of aspects 1 to 14, wherein the amylin analogue is administered to the subject at a dose of about 0.04 mg to about 10 mg, for example about 0.04 mg to about 7 mg, about 0.04 mg to about 6 mg, about 0.04 mg to about 4.4 mg, about 0.04 mg to about 2.4 mg, about 0.6 mg to about 6 mg, about 0.6 mg to about 4.4 mg, about 0.6 mg to about 2.4 mg, about 0.7 mg to about 6 mg, about 0.7 mg to about 4.4 mg, about 0.7 mg to about 2.4 mg.

[0725] Aspect 16. The dosing regimen, the method, the amylin analogue for use, or the use of any one of aspects 1 to 15, wherein the amylin analogue is administered to the subject at a dose of about 10 mg, about 9.5 mg, about 9 mg, about 8.5 mg, about 8 mg, about 7.5 mg, about 7 mg, about 6.5 mg, about 6 mg, about 4.4 mg, about 3.4 mg, about 2.4 mg, about 1.4 mg, about 0.7 mg, about 0.35 mg, about 0.16 mg, about 0.08 mg, and / or about 0.04 mg.

[0726] Aspect 17. The dosing regimen, the method, the amylin analogue for use, or the use of any one of aspects 1 to 16, wherein the amylin analogue is administered to the subject at a fixed interval.

[0727] Aspect 18. The dosing regimen, the method, the amylin analogue for use, or the use of any one of aspects 1 to 17, wherein the amylin analogue is administered to the subject once a day, once every two days, once a week, once every two weeks, once every three weeks, or once every four weeks.

[0728] Aspect 19. The dosing regimen, the method, the amylin analog for use, or the use of any one of aspects 1 to 18, wherein the dose of the amylin analog is the same at each administration.

[0729] Aspect 20. The dosing regimen, the method, the amylin analog for use, or the use of any one of aspects 1 to 18, wherein the dose of the amylin analog is different at each administration.

[0730] Aspect 21. The dosing regimen, the method, the amylin analog for use, or the use of any one of aspects 1 to 20, wherein the amylin analog is administered intravenously.

[0731] Aspect 22. The dosing regimen, the method, the amylin analog for use, or the use of any one of aspects 1 to 20, wherein the amylin analog is administered subcutaneously.

[0732] Aspect 23. The dosing regimen, the method, the amylin analog for use, or the use of any one of aspects 1 to 22, wherein the activity of the concomitant agent is affected by the rate of gastric emptying.

[0733] Aspect 24. The dosing regimen, the method, the amylin analog for use, or the use of any one of aspects 1 to 23, wherein the administration of the concomitant agent is affected by or depends on the rate of gastric emptying.

[0734] Aspect 25. The dosing regimen, the method, the amylin analog for use, or the use of any one of aspects 1 to 24, wherein the dose of the administered concomitant agent is affected by or depends on the rate of gastric emptying.

[0735] Aspect 26. The dosing regimen, the method, the amylin analog for use, or the use of any one of aspects 1 to 25, wherein the concomitant agent is an oral drug.

[0736] Aspect 27. The dosing regimen, the method, the amylin analog for use, or the use of any one of aspects 1 to 26, wherein the concomitant agent is a non-oral drug.

[0737] Aspect 28. The dosing regimen, the method, the amylin analog for use, or the use of any one of aspects 1 to 29, wherein the concomitant agent is selected from the group consisting of: analgesics, antibiotics, anticoagulants, contraceptives, antihypotensive drugs, statins, antifungal drugs, hypoglycemic agents, contraceptives, oral drugs with narrow therapeutic index, and nutritional supplements.

[0738] Aspect 29. The dosing regimen, the method, the amylin analogue for use, or the use of any one of aspects 1 to 28, wherein the concomitant agent is selected from the group consisting of: insulin, acetaminophen, digoxin, warfarin, metformin, hydrocortisone, griseofulvin, glibenclamide, glicazide, and glipizide.

[0739] Aspect 30. The dosing regimen, the method, the amylin analogue for use, or the use of any one of aspects 1 to 29, wherein the concomitant agent is insulin.

[0740] Aspect 31. The dosing regimen, the method, the amylin analogue for use, or the use of any one of aspects 1 to 30, wherein the subject is a human subject.

[0741] Aspect 32. A dosing regimen for a subject, wherein the dosing regimen comprises:

[0742] (a) administering to the subject an amylin analogue that does not affect gastric emptying rate; and

[0743] (b) administering to the subject a concomitant agent.

[0744] Aspect 33. A method of treating or preventing a disease in a subject, wherein the method comprises:

[0745] (a) administering to the subject an amylin analogue that does not affect gastric emptying rate; and

[0746] (b) administering to the subject a concomitant agent.

[0747] Aspect 34. An amylin analogue that does not affect gastric emptying rate for use in a method of treating or preventing a disease in a subject, wherein the method comprises:

[0748] (a) administering to the subject the amylin analogue; and

[0749] (b) administering to the subject a concomitant agent.

[0750] Aspect 35. Use of an amylin analogue that does not affect gastric emptying rate for the manufacture of a medicament for use in a method of treating or preventing a disease in a subject, wherein the method comprises:

[0751] (a) administering to the subject the amylin analogue; and

[0752] (b) administering to the subject a concomitant agent.

[0753] Aspect 36. A method of inhibiting weight gain and / or reducing body weight in a subject, wherein the method comprises:

[0754] (a) administering to the subject an amylin analogue that does not affect gastric emptying rate; and

[0755] (b) administering to the subject a concomitant agent.

[0756] Aspect 37. The aspects of any one of aspects 32 to 36, comprising the features of any one or more of aspects 1 to 31.

[0757] Examples

[0758] The following examples are provided to illustrate some preferred aspects of the application and are not intended to limit the scope of the application.

[0759] Materials and methods

[0760] Mixed test meal (MTM)

[0761] In the single dose > 0.7 mg dose cohort, the same Mixed Test Meal (MTM) was performed on Day -1 and Day 5 (Example 1). In the multiple dose > 0.6 mg dose cohort, the same Mixed Test Meal (MTM) was performed on Day -1 and Day 5, and Day 40 (Example 2).

[0762] A population predetermined MTM (see below) was provided for breakfast, which was standardized according to average subject characteristics (two BMI categories were used). This meal consisted of fixed nutrient content.

[0763] The start of the intake of the liquid component of the meal together with the paracetamol tablets was defined as time point 0 (t=0).

[0764] The subjects were required to be in a fasted state prior to the test. A fixed composition meal was used as the MTM and was consumed by the subjects in a predetermined order within 15 minutes of time point 0 (zero).

[0765] During the intake of the mixed meal, the subjects were allowed to sit.

[0766] The subjects were advised to avoid eating until after the meal test period (240 minutes).

[0767] In addition to the water provided with the test meal, no water was allowed to be consumed during the first 2 hours after the start of the meal intake, after which the subjects were allowed to drink water ad libitum.

[0768] Subjects were required to lie in semi-recumbent position for at least 4 hours after MTM / acetaminophen administration and in supine or semi-recumbent position thereafter until the last blood sample was taken according to the blood sampling schedule (see Table 1 below) to determine post-prandial glucose (PPG), insulin, glucagon and acetaminophen concentrations.

[0769] Acetaminophen concentration was used as a marker for post-prandial gastric emptying.

[0770] After the end of the meal test procedure, subjects were offered a meal. In case of early termination of each meal test, the reason for termination was recorded in the case report form.

[0771] Table 1 : Blood sampling schedule

[0772]

[0773] 1 Samples for acetaminophen, plasma glucose, insulin and glucagon were taken within the following time windows:

[0774] t=0 (pre-ingestion) -30 minutes, and time windows: t=15 to t=120 minutes ± 2 minutes; t=180 and 240 minutes ± 5 minutes.

[0775] 2 The nominal time point t=0 minutes was defined as the start of ingestion of the liquid component of the MTM with acetaminophen (2x500 mg). The test meal should be consumed within 15 minutes.

[0776] Table 2: Composition of the standardized test meal for subjects with BMI: > 21.0 to < 25.0 kg / m2

[0777] 30% of total energy expenditure

[0778]

[0779] The ingredients of the mixed meal are exemplary and can vary according to the subject's taste, aversion or allergy, but the relationship of the nutrients is the same.

[0780] Table 3: Composition of the standardized test meal for subjects with BMI: > 25.0 to 29.9 kg / m2

[0781] 30% of total energy expenditure

[0782]

[0783] The ingredients of the mixed meal are exemplary and can vary according to the subject's taste, aversion or allergy, but the relationship of the nutrients is the same.

[0784] Paracetamol absorption test

[0785] Two 500 mg acetaminophen tablets (total 1000 mg) were administered with the scheduled solid MTM. Subjects were instructed to take the tablets with the liquid component of the meal at the start of the meal test. Frequent blood sampling for acetaminophen concentration was performed as described above.

[0786] Example 1 : Phase 1a clinical trial to assess single dose escalation

[0787] A first-in-human, randomized, single ascending dose (SAD) trial to assess the safety, tolerability, pharmacokinetics, and pharmacodynamics of the amylin analogue of SEQ ID NO: 3 administered to healthy subjects was performed.

[0788] Subjects were healthy male subjects with normal weight and overweight (BMI: 21.0 to 29.9 kg / m 2 ).

[0789] Trial design

[0790] The trial was a single center, randomized, double-blind, placebo-controlled, single ascending dose trial in male subjects with normal weight and overweight who were otherwise healthy, randomized within each cohort to the amylin analogue of SEQ ID NO: 3 or placebo (randomization ratio 3: 1; n = 6 and n = 2). Fifty-six subjects were assigned to the following seven ascending dose levels: 0.04, 0.08, 0.16, 0.35, 0.7, 1.4, 2.4 mg, administered as a subcutaneous (s.c.) dose, as shown in Figure 1 Eight subjects were assigned to an i.v. cohort at one dose level of 0.35 mg.

[0791] A sentinel dosing approach (sequential dosing) was applied within each cohort. The overall observation period comprised 50 days, starting with an in-house stay (Day -1 to Day 8), discharge on Day 8, followed by 6 outpatient visits, and an End of Trial (EOT) visit on Day 50.

[0792] Blinded evaluation of each cohort was performed by the Trial Safety Group (TSG) to determine whether the trial should progress to the next dose level based on the specified stopping rules applied to the trial.

[0793] Formulation of the amylin analogue of SEQ ID NO: 3 (4 mg / mL) and placebo is shown in Table 4 below.

[0794] Table 4: Formulation parameters

[0795]

[0796] Preparation of the drug product and matching placebo was performed by Rechon Life Science AB, Limhamn, Sweden on behalf of Zealand Pharma A / S. Secondary packaging and labelling of the drug product and matching placebo was performed by Klifo A / S, Glostrup, Denmark.

[0797] The subjects received a single dose consisting of 0.04 mg, 0.08 mg, 0.16 mg, 0.35 mg, 0.7 mg, 1.4 mg or 2.4 mg of the amylin analogue or placebo administered subcutaneously or 0.35 mg administered intravenously, as shown in the table below.

[0798] Table 5: Dose and cohort parameters

[0799]

[0800] All demographics were balanced in the healthy subject cohort.

[0801] The baseline characteristics of the subjects are shown below.

[0802] Table 6: Subject parameters

[0803]

[0804] The subjects were tested according to the mixed meal test and the paracetamol absorption test described in the methods.

[0805] Results

[0806] The subjects receiving the amylin analogue exhibited a dose-dependent and sustained reduction in body weight ( Figure 3 ).

[0807] The results from the mixed meal test show that the subjects administered the amylin analogue herein exhibited a dose-dependent inhibition of glucagon secretion ( Figure 4 ). Amylin is known to affect glucagon secretion, inhibiting its secretion in response to amino acid stimulation and reducing endogenous glucose production during the postprandial phase.

[0808] Unexpectedly, no dose relationship was observed between the administered amylin analogue and paracetamol absorption ( Figure 5 and Table 7).

[0809] The amylin analog did not actually change both the maximum concentration (Cmax) of acetaminophen and the time it took to reach that maximum concentration (Tmax) in the acetaminophen absorption test compared to placebo.

[0810] Table 7: Pharmacokinetic parameters of paracetamol in the paracetamol absorption test

[0811]

[0812] Thus, these data indicate that the amylin analog did not change the rate of gastric emptying at all doses relative to placebo.

[0813] The amylin analog was well tolerated, with no serious or severe adverse events (AEs) and no withdrawals.

[0814] The most frequent AEs were decreased appetite, nausea, and vomiting; most AEs were mild and transient.

[0815] In addition, no anti-drug antibodies were detected.

[0816] Table 8: Adverse events (AEs)

[0817]

[0818] All events meeting the definition of an AE were collected and reported from the first trial-related activity after the subject signed the informed consent and until the end of the post-treatment follow-up period. At each site contact (visit or phone, excluding safety visits where the subject did not see the investigator or site staff, e.g., to visit the laboratory), the subject was asked about AEs. All AEs observed by the investigator or reported by the subject were recorded and evaluated by the investigator.

[0819] The investigator recorded the diagnosis, if possible. If a diagnosis could not be made, the investigator recorded each sign and symptom as a separate AE.

[0820] The maximum intensity (severity) of all AEs was assessed and recorded by the investigator.

[0821] In the case of known AE outcomes, severity was graded according to the following:

[0822] Mild: Type of adverse event that is usually transient and can require only minimal treatment or therapeutic intervention. The event generally does not interfere with normal activities of daily life.

[0823] Moderate: Type of adverse event that usually lessens with additional specific therapeutic intervention. The event interferes with normal activities of daily life, causes discomfort, but does not pose a significant or permanent hazard to the study participant.

[0824] Severe: Type of adverse event that interrupts normal activities of daily life, or significantly affects clinical status or can require intensive therapeutic intervention. A "severe" reaction does not necessarily mean that the AE is "serious", and an SAE can not be "severe" in nature.

[0825] The observation of a sustained dose-dependent reduction in body weight indicates that the amylin analog is suitable for the treatment of obesity or for aiding weight loss.

[0826] Furthermore, the unexpected lack of effect on gastric emptying can facilitate dosing regimens and concomitant dosing that are not recommended or are less effective for other amylin analogs.

[0827] Summary of Example 1

[0828] The amylin analog of the present invention is a long-acting amylin analog originally designed for once-weekly dosing that shows potential for reducing body weight and improving glycemia in animal models of obesity and diabetes.

[0829] The purpose of the present example is to provide a first-in-human trial to evaluate the safety, tolerability, pharmacokinetics (PK) and pharmacodynamics (PD) of the amylin analog of SEQ ID NO: 3, with the primary endpoint being adverse events (AEs) and the secondary endpoints being PL parameters of the amylin analog and PD parameters related to the mixed test meal (MTM).

[0830] The amylin analog of SEQ ID NO: 3 can be referred to as ZP8396.

[0831] The above trial is a randomized, double-blind, placebo-controlled trial of a single subcutaneous injection of the amylin analog of SEQ ID NO: 3 in healthy, lean and overweight male subjects to assess its safety, PK and PD. A total of 56 subjects (mean age 38.1 years; mean BMI 25.6 kg / m 2 ) were randomized to amylin analog or placebo (6:2) within seven dose cohorts of 0.04 to 2.4 mg.

[0832] After 1 week of observation, mean body weight decreased by -0.6%, 2.6%, 3.6%, and 4.2% relative to baseline after single doses of placebo, 0.7, 1.4, and 2.4 mg, respectively. The amylin analog was well tolerated, with no serious or severe AEs, and no withdrawals. The number and severity of gastrointestinal AEs increased with dose; the mean half-life of the amylin analog was about 10 days Figure 2 ), and a dose-dependent decrease in glucagon release was observed Figure 4 ). Furthermore, no anti-drug antibodies were detected.

[0833] The data show that the amylin analog was well tolerated at single doses up to 2.4 mg. The most common related AEs were decreased appetite, nausea, and vomiting, however most events were mild and transient (Table 8). The half-life of about 10 days is suitable for once-weekly dosing. Furthermore, single doses of the amylin analog resulted in a dose-dependent and sustained decrease in body weight Figure 2 ), which supports the potential as a treatment for obesity.

[0834] Importantly, the data in this Example indicate that the amylin analog (ZP8396) at all tested doses did not change the rate of gastric emptying relative to placebo.

[0835] Example 2: Phase 1 clinical trial (Part 1) to evaluate multiple dose escalation

[0836] A first-in-human, randomized, multiple-dose escalation (MAD) trial was conducted to evaluate the safety, tolerability, pharmacokinetics, and pharmacodynamics of the amylin analog of SEQ ID NO: 3 administered to healthy subjects.

[0837] Subjects were healthy male subjects with normal weight and overweight (BMI: 21.0 to 29.9 kg / m2).

[0838] Trial design

[0839] The trial was a single-center, within-group randomized and double-blind, placebo-controlled, sequential multiple-dose escalation trial in normal weight and overweight, otherwise healthy subjects randomized to the amylin analog of SEQ ID NO: 3 or placebo (randomization ratio 7:3) within each cohort. Subjects were assigned to two escalating dose levels: 6 weekly administrations of the amylin analog or placebo at 0.6 or 1.2 mg, administered with subcutaneous (s.c.) dosing, as shown in Figure 6 .

[0840] The sentinel dosing approach (sequential dosing) was applied within cohorts. The entire observation period comprised 92 days, starting with the first in-patient stay (Day -1 to Day 2), planned discharge on Day 2, followed by one outpatient visit (Day 5); second in-patient stay (Day 8 to Day 12), planned discharge on Day 12; third in-patient stay (Day 15 to Day 19), planned discharge on Day 19; fourth in-patient stay (Day 22 to Day 24), planned discharge on Day 24, followed by one outpatient visit (Day 26); fifth in-patient stay (Day 29 to Day 31), planned discharge on Day 31; sixth in-patient stay (Day 36 to Day 38), planned discharge on Day 38, followed by five outpatient visits (Day 40, Day 43, Day 50, Day 64, and Day 75), and the End of Trial (EOT) visit on Day 92.

[0841] Blinded evaluation of each cohort was performed by the Trial Safety Group (TSG) to determine if the trial should progress to the next dose level based on the specified stopping rules applied to the trial.

[0842] Formulation of amylin analogue of SEQ ID NO: 3 (4 mg / mL) and placebo is shown in Table 9 below.

[0843] Table 9: Formulation parameters

[0844]

[0845] Preparation of the drug product and matching placebo was performed by Rechon Life Science AB, Limhamn, Sweden on behalf of Zealand Pharma A / S. Klifo A / S, Glostrup, Denmark was responsible for secondary packaging and labelling of the drug product and matching placebo.

[0846] As shown in the table below, subjects received a single dose consisting of subcutaneous administration of 0.6 mg or 1.2 mg amylin analogue or placebo once weekly.

[0847] Table 10: Dose and cohort parameters

[0848]

[0849] All demographics were balanced in the healthy subject cohort. Baseline characteristics of the subjects are shown in Table 11 below.

[0850] Table 11 : Subject parameters

[0851]

[0852] 20 subjects with an average body weight of 82 kg and a BMI of 25.4 kg / m 2 participated in the trial. Two subjects did not receive the 6thdose. All 20 randomized subjects completed the trial.

[0853] Subjects were tested according to the mixed meal test and paracetamol absorption test described in the "Materials and Methods" section herein.

[0854] Results

[0855] According to the trial design as detailed above, data related to consecutive dose cohorts (i.e. cohorts receiving an increasing dose relative to the previous cohort) could be obtained sequentially.

[0856] Initial data indicated a favorable outcome consistent with the SAD trial.

[0857] Subjects receiving the amylin analog showed a body weight reduction of 0.4%, 5.3% and 5.1% relative to baseline after 6 administrations of placebo, 0.6 mg and 1.2 mg once weekly, respectively. Figure 7 Individual and mean (thick line) body weight loss during the trial is shown.

[0858] Table 12: Mean body weight one week after the sixth dose of placebo or the amylin analogue of SEQ ID NO: 3 Change

[0859]

[0860] Results from the mixed meal test further indicated that subjects administered with the amylin analog herein showed no significant difference in paracetamol concentration after a single administration (day 5) or six administrations (day 40) compared to pre-dose or placebo (Table x and Figure 8 ).

[0861] Table 13: Pharmacokinetic parameters of paracetamol in the paracetamol absorption test

[0862]

[0863] Results from the mixed test meal indicated that subjects administered with the amylin analog herein showed a significant difference in glucagon concentration relative to baseline at day 5 compared to placebo; the high variability in glucagon concentration within subjects at day 40 resulted in no difference shown compared to placebo ( Figure 9 ). Amylin is known to affect glucagon secretion, inhibiting its secretion by amino acid stimulation and reducing endogenous glucose production during the postprandial phase.

[0864] The amylin analog was well tolerated, with no serious or severe adverse events (AEs) and no withdrawals.

[0865] Table 14: Adverse events (AEs)

[0866]

[0867] Summary of Example 2

[0868] Amylin analogs of SEQ ID NO: 3 designed for once-weekly dosing showed potential to reduce body weight and improve glycemia in obese and diabetic animal models. Dose-dependent and sustained reductions in body weight were shown in healthy subjects following a single dose of amylin analog, supporting the potential of amylin analogs as a treatment for obesity.

[0869] A first part of a randomized, double-blind, placebo-controlled trial designed to assess the safety, pharmacokinetics, and pharmacodynamics of amylin analog of SEQ ID NO: 3 administered once-weekly for 6 subcutaneous injections in healthy lean and overweight subjects has been completed. A total of 20 subjects (with a mean body weight of 82 kg and a mean BMI of 25.4 kg / m2) were randomized to amylin analog of SEQ ID NO: 3 or placebo (7:3) within two dose cohorts. 2 ) were randomized to amylin analog of SEQ ID NO: 3 or placebo (7:3) within two dose cohorts.

[0870] Mean body weight decreased by 0.4%, 5.3%, and 5.1% relative to baseline following once-weekly for 6 doses of placebo, 0.6 mg, and 1.2 mg, respectively.

[0871] The amylin analog of SEQ ID NO: 3 was well tolerated, with no serious or severe adverse events (AEs) and no withdrawals. The most frequent related AEs were decreased appetite, early satiety, food aversion, and nausea, all of which were mild and transient. Only three subjects treated with amylin analog experienced nausea, one of whom also reported vomiting. No injection site reactions were reported, and no anti-drug antibodies were detected.

[0872] Treatment with six doses of amylin analog of SEQ ID NO: 3 was safe and well tolerated and resulted in a significant reduction in body weight. The most common related AEs were related to the GI system, all of which were mild and most of which occurred within 2 days of the first dose. These data confirm the observation of body weight loss following a single dose of amylin analog and are similar to the body weight loss observed following 6 weeks of treatment with other weight loss therapies. Cohort studies exploring amylin analog doses higher than 1.2 mg with longer treatment durations and dose escalation titrations are ongoing to further assess the clinical potential of amylin analog of SEQ ID NO: 3.

[0873] Importantly, the data in this example indicate that the amylin analogue (ZP8396) at all tested doses does not change the rate of gastric emptying relative to placebo.

[0874] Example 3: Phase 1 clinical trial (Part 2) to assess multiple dose escalation

[0875] A randomized, multiple dose escalation trial to assess the safety, tolerability, pharmacokinetics and pharmacodynamics of the amylin analogue of SEQ ID NO: 3 administered to healthy subjects.

[0876] Subjects are healthy male or non-childbearing potential female subjects who are overweight and obese but otherwise healthy, and have a body mass index (BMI) of 27.0 to 39.9 kg / m2 2 .

[0877] Trial design

[0878] Part 2 of the MAD trial is a single center, within-group randomized and double-blind, placebo-controlled, sequential multiple dose escalation trial in overweight and obese but otherwise healthy subjects randomized within each cohort to the amylin analogue of SEQ ID NO: 3 or placebo (randomization ratio 3: 1). The use of a dose escalation titration will be explored to reach higher exposures of the amylin analogue of SEQ ID NO: 3.

[0879] For a given cohort, steady state is reached after 6 dosing maintenance doses of the amylin analogue of SEQ ID NO: 3 once weekly. Subjects will be assigned to three cohorts with the objective of three different maintenance doses, where 16 administrations of the amylin analogue of SEQ ID NO: 3 or placebo (as shown) are administered s.c. once weekly: Figure 10

[0880] 1. Cohort 1 : 0.6, 0.6, 1.2, 1.2 mg followed by a maintenance dose of 2.4 mg for 12 weeks.

[0881] 2. Cohort 2: 0.6, 0.6, 1.2, 1.2, 2.4, 2.4, 3.6, 3.6 mg followed by a maintenance dose of 4.8 mg for 8 weeks.

[0882] 3. Cohort 3: 0.6, 0.6, 1.2, 1.2, 2.4, 2.4, 3.6, 3.6, 4.8, 4.8 mg followed by a maintenance dose of 6.0 mg for 6 weeks.

[0883] ​Forty-eight subjects were randomized to three groups of 16 subjects, in each group the subjects were in a 3: 1 ratio, in which 12 subjects received the amylin analogue of SEQ ID NO: 3 and 4 subjects received placebo.

[0884] The total observation period was 25 weeks, starting with the first inpatient stay (Day -1 to Day 2), planned discharge on Day 2, followed by 3 outpatient visits once weekly dosing, and followed by 13 inpatient visits according to dose level, with overnight stay of 1 to 4 days. After 16 weekly doses (Day 1, 8, 15, 22, 29, 36, 43, 50, 57, 64, 71, 78, 85, 92, 99, and 106), Group 1 had six outpatient visits (Day 108, 110, 113 ± 1, 120 ± 1, 134 ± 1, 148 ± 1), and Groups 2 and 3 had five outpatient visits (Day 110, 113 ± 1, 120 ± 1, 134 ± 1, 148 ± 1), and the trial ended with an End of Trial (EOT) visit on Day 169 ± 2. Blinded evaluation of each group was performed by the Trial Safety Group (TSG), with data cut-off prior to the assessment of the 5th dose of maintenance dosing (Day 57 for Group 1, and Day 85 for Group 2, and Day 99 for Group 3) to determine if the current dose level was considered safe and if the dose escalation could continue in the next group based on the stopping rules.

[0885] Formulation of the amylin analogue of SEQ ID NO: 3 (4 mg / mL) and placebo is shown in Table 15 below.

[0886] Table 15: Formulation parameters

[0887]

[0888]

[0889] Preparation of the drug product and matching placebo above was performed by Rechon Life Science AB, Limhamn, Sweden on behalf of Zealand Pharma A / S. Secondary packaging and labelling of the drug product and matching placebo was performed by Klifo A / S, Glostrup, Denmark.

[0890] Subjects were tested on Day -1 and Day 110 according to the Mixed Meal Test and Acetaminophen Absorption Test described in the Materials and Methods section herein.

[0891] Results

[0892] According to the trial design as detailed above, data relating to successive dose cohorts (i.e. cohorts receiving an increasing dose relative to the previous cohort) can be obtained sequentially.

[0893] The initial data indicate a favourable outcome, which is generally consistent with the SAD trial in Example 1 herein and Part 1 of the MAD trial in Example 2 herein.

[0894] All publications mentioned in the above specification are herein incorporated by reference. Various modifications and variations of the described methods and systems of the application will be apparent to those skilled in the art without departing from the scope and spirit of the application. Although the application has been described in connection with specific preferred embodiments, it should be understood that the application as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes of carrying out the application that are obvious to those skilled in biochemistry, molecular biology or related fields are intended to be within the scope of the following claims.

Claims

1. An amylin analogue that does not affect gastric emptying rate for use in a method of treating or preventing a disease in a subject, wherein the method comprises: (a) administering the amylin analogue to the subject; and (b) administering a concomitant agent to the subject; wherein: the amylin analogue and the concomitant agent are administered to the subject within about 120 minutes of each other; and the amylin analogue comprises or consists of a peptide of the formula: or a pharmaceutically acceptable salt and / or derivative thereof; wherein: R 1 is hydrogen, C 1-4 acyl, benzoyl or C 1-4 alkyl, or a half-life extending moiety M, wherein M is optionally connected to Z via a linker moiety L; R 2 is OH or NHR 3 wherein R 3 is hydrogen or C 1-3 alkyl; and Z is an amino acid sequence of Formula I: wherein: X1 is selected from Arg, Lys and Glu; X3 is selected from Gly, Gin and Pro; X4 is selected from Thr and Glu; X5 is selected from Ala and Leu; X6 is selected from Thr and Ser; X10 is selected from Glu and Gin; X14 is selected from Aad, His, Asp, Asn and Arg; X17 is selected from Gin, His and Thr; X19-X20 is selected from Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly and Ala-Asn, or is absent; X27 is selected from Leu and Pro; X32 is selected from Val and Thr; X35 is selected from Asn and Ser; X37 is selected from Hyp and Pro; and X2 and X7 are amino acid residues whose side chains together form a lactam bridge; Gly(Me) is N-methylglycine [also known as sarcosine (Sar)] Ile(Me) is N-methylisoleucine Aad is 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2-amino hexanedioic acid], also known as homoglutamic acid.

2. A non-therapeutic dosing regimen for a subject, wherein the dosing regimen comprises: (a) administering an amylin analogue that does not affect gastric emptying rate to the subject; and (b) administering a concomitant agent to the subject; wherein: the amylin analogue and the concomitant agent are administered to the subject within about 120 minutes of each other; and the amylin analogue comprises or consists of a peptide of the formula: or a pharmaceutically acceptable salt and / or derivative thereof; wherein: R 1 is hydrogen, C 1-4 acyl, benzoyl or C 1-4 alkyl, or a half-life extending moiety M, wherein M is optionally connected to Z via a linker moiety L; R 2 is OH or NHR 3 wherein R 3 is hydrogen or C 1-3 alkyl; and Z is an amino acid sequence of Formula I: wherein: X1 is selected from Arg, Lys and Glu; X3 is selected from Gly, Gin and Pro; X4 is selected from Thr and Glu; X5 is selected from Ala and Leu; X6 is selected from Thr and Ser; X10 is selected from Glu and Gin; X14 is selected from Aad, His, Asp, Asn and Arg; X17 is selected from Gin, His and Thr; X19-X20 is selected from Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly and Ala-Asn, or is absent; X27 is selected from Leu and Pro; X32 is selected from Val and Thr; X35 is selected from Asn and Ser; X37 is selected from Hyp and Pro; and X2 and X7 are amino acid residues whose side chains together form a lactam bridge; Gly(Me) is N-methylglycine [also known as sarcosine (Sar)] Ile(Me) is N-methylisoleucine Aad is 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2-amino hexanedioic acid], also known as homoglutamic acid.

3. A non-therapeutic method of inhibiting weight gain and / or reducing body weight in a subject, wherein the method comprises: (a) administering to the subject an amylin analogue that does not affect gastric emptying rate; and (b) administering to the subject a concomitant agent; wherein: the amylin analogue and the concomitant agent are administered to the subject within about 120 minutes of each other; and the amylin analogue comprises or consists of a peptide of the formula: wherein: R 1 is hydrogen, C 1-4 acyl, benzoyl or C 1-4 alkyl, or a half-life extending moiety M, wherein M is optionally linked to Z via a linker moiety L; R 2 is OH or NHR 3 wherein R 3 is hydrogen or C 1-3 alkyl; and Z is an amino acid sequence of the formula I: wherein: X1 is selected from Arg, Lys and Glu; X3 is selected from Gly, Gin and Pro; X4 is selected from Thr and Glu; X5 is selected from Ala and Leu; X6 is selected from Thr and Ser; X10 is selected from Glu and Gin; X14 is selected from Aad, His, Asp, Asn and Arg; X17 is selected from Gin, His and Thr; X19-X20 is selected from Ser-Ser, Thr-Thr, Ala-Thr, Ala-Ala, Gly-Thr, Gly-Gly and Ala-Asn, or is absent; X27 is selected from Leu and Pro; X32 is selected from Val and Thr; X35 is selected from Asn and Ser; X37 is selected from Hyp and Pro; and X2 and X7 are amino acid residues whose side chains together form a lactam bridge; Gly(Me) is N-methylglycine [also known as sarcosine (Sar)] Ile(Me) is N-methylisoleucine Aad is 2-amino adipic acid, for example (2S)-2-amino adipic acid [also known as (2S)-2-amino hexanedioic acid], also known as homoglutamic acid.

4. The dosing regimen of claim 2 or the method of claim 3, wherein the subject is overweight, obese or morbidly obese.

5. The amylin analogue for use of claim 1, wherein the disease is overweight, obesity, morbid obesity, diabetes, or a disease associated with obesity or diabetes.

6. The amylin analogue for use according to claim 5, wherein the disease associated with obesity or diabetes is selected from the group consisting of obesity-associated inflammation, obesity-associated gallbladder disease, obesity-induced sleep apnea, obesity-associated respiratory problems, cartilage degeneration, osteoarthritis, infertility, Alzheimer's disease, prediabetes, insulin resistance syndrome, impaired glucose tolerance (IGT), a disease state associated with elevated blood glucose levels, metabolic disease, metabolic syndrome, hyperglycemia, hypertension, atherogenic dyslipidemia, hepatic steatosis, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), renal failure, arteriosclerosis, macrovascular disease, microvascular disease, diabetic heart disease, diabetic cardiomyopathy, heart failure as a complication of diabetes, coronary heart disease, peripheral arterial disease, and stroke.

7. The amylin analogue for use according to any one of claims 1, 5 and 6, the dosage regimen according to claim 2 or claim 4, or the method according to claim 3 or claim 4, wherein the amylin analogue comprises or consists of a peptide of the formula: ###0001### or a pharmaceutically acceptable salt and / or derivative thereof. 。 8. The amylin analogue for use, the dosage regimen or the method according to claim 7, wherein the amylin analogue is petrelintide, which is a compound of the formula: ###0002### or a pharmaceutically acceptable salt thereof. wherein an intramolecular lactam bridge is formed between the side chains of the residues indicated by brackets "()"; and wherein [19CD]-isoGlu is a 19-carboxy nonadecanoyl group covalently attached to the alpha amino group of the isoGlu linker.

9. The amylin analogue for use, the dosage regimen or the method according to any one of claims 1 to 8, wherein the amylin analogue and the concomitant agent are administered to the subject within about 90 minutes of each other, within about 60 minutes of each other, within about 45 minutes of each other, within about 30 minutes of each other, within about 15 minutes of each other and / or substantially simultaneously with each other.

10. The amylin analogue for use, the dosage regimen or the method according to any one of claims 1 to 9, wherein the amylin analogue is administered to the subject in a dose of about 0.04 mg to about 10 mg, for example about 0.04 mg to 7 mg, about 0.04 mg to about 6 mg, about 0.04 mg to about 4.4 mg, about 0.04 mg to about 2.4 mg, about 0.6 mg to about 6 mg, about 0.6 mg to about 4.4 mg, about 0.6 mg to about 2.4 mg, about 0.7 mg to about 6 mg, about 0.7 mg to about 4.4 mg, about 0.7 mg to about 2.4 mg.

11. The amylin analogue for use, the dosing regimen or the method of any one of claims 1 to 10, wherein the amylin analogue is administered to the subject at a dose of about 10 mg, about 9.5 mg, about 9 mg, about 8.5 mg, about 8 mg, about 7.5 mg, about 7 mg, about 6.5 mg, about 6 mg, about 4.4 mg, about 3.4 mg, about 2.4 mg, about 1.4 mg, about 0.7 mg, about 0.35 mg, about 0.16 mg, about 0.08 mg and / or about 0.04 mg.

12. The amylin analogue for use, the dosing regimen or the method of any one of claims 1 to 11, wherein the amylin analogue is administered to the subject once a day, once every two days, once a week, once every two weeks, once every three weeks or once every four weeks.

13. The amylin analogue for use, the dosing regimen or the method of any one of claims 1 to 12, wherein: (i) the activity of the concomitant agent is influenced by the rate of gastric emptying; (ii) the timing of administration of the concomitant agent is influenced by or dependent on the rate of gastric emptying; and / or (iii) the dose of the concomitant agent administered is influenced by or dependent on the rate of gastric emptying.

14. The amylin analogue for use, the dosing regimen or the method of any one of claims 1 to 13, wherein the concomitant agent is an oral or a parenteral drug.

15. The amylin analogue for use, the dosing regimen or the method of any one of claims 1 to 14, wherein the concomitant agent is a drug selected from the group consisting of analgesics, antibiotics, anticoagulants, contraceptives, antihypotensive drugs, statins, antifungal drugs, hypoglycemic agents, oral drugs with narrow therapeutic index and nutritional supplements.

16. The amylin analogue for use, the dosing regimen or the method of any one of claims 1 to 15, wherein the concomitant agent is selected from the group consisting of insulin, acetaminophen, digoxin, warfarin, metformin, hydrocortisone, griseofulvin, glibenclamide, gliclazide and glipizide.

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