EA042757B1 - ANALOGUES OF AMILIN - Google Patents

Description

The present application claims priority on the basis of European Patent Application No.

16188024.0, filed September 9, 2016, the contents of which are incorporated herein by reference in their entirety.

Technical field

The present invention relates to amylin analogs, which are amylin receptor agonists, and their medical use in the treatment and/or prevention of a number of diseases, conditions or disorders, including the treatment and/or prevention of excessive food intake, obesity and overweight, metabolic diseases, and others. conditions and disorders described in the present description. In particular, the present invention relates to stable amylin analogues that have a long duration of action and are well suited for use in the form of a liquid formulation.

State of the art

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

Native human amylin is a 37 amino acid peptide having the formula

H-KC()NTATC()ATQRLANFLVHSSNNFGAILSSTNVGSNTY-NH ₂ , where H- at the N-terminus denotes a hydrogen atom, which corresponds to the presence of a free amino group at the N-terminal amino acid residue [i.e., the lysine (K) residue at position number 1 of the sequence represented higher]; where -NH2 at the C-terminus indicates that the C-terminal carboxyl group is in the amide form; and where parentheses () at two cysteine residues (C, Cys) at positions 2 and 7 of the sequence indicate the presence of an intramolecular disulfide bridge between the two Cys residues in question.

Amylin may be suitable for use in the treatment of metabolic disorders such as diabetes and/or obesity. Amylin is believed to regulate gastric emptying and suppress glucagon secretion and food intake, thus regulating the rate of glucose release into the bloodstream. Amylin appears to complement the action of insulin. Compared with healthy adults, patients with type 1 diabetes do not have circulating amylin, and patients with type 2 diabetes show a decrease in amylin concentrations in the postprandial period. In human studies, it has been shown that the amylin analog known as pramlintide described in WO 93/10146 and having the sequence Lys-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala- Asn-Phe-Leu-Val-His-Ser-SerAsn-Asn-Phe-Gly-Pro-lle-Leu-Pro-Pro-Thr-Asn-Val-Gly-Ser-Asn-Thr-Tyr, which also contains a disulfide bridge between Cys residues in positions 2 and 7, provides weight loss or limits weight gain. An alternative amylin analog containing N-methylated residues and having a reduced propensity to fibril, designated IAPP-GI, has been described by Yan et al. (PNAS, 103(7), 2046-2051, 2006; Angew. Chem. Int. Ed. 2013, 52, 10378-10383; WO 2006/042745). However, IAPP-GI appeared to be less effective than native amylin.

WO 91/07978 describes analogues of hypocalcemic peptides, including amylin, in which the internal disulfide bridges are replaced with alternative ring structures. The effect of these alternative structures on the activity of amylin analogs has not been disclosed. WO 99/34764 presents data showing that ^2,7 cyclo-[2Asp,7Lys]-h-amylin has significantly lower activity than some other amylin analogs and human amylin itself.

Other analogs of amylin or pramlintide are described in WO 2013/156594, WO 2012/168430, WO 2012/168431 and WO 2012/168432, as well as WO 2015/040182.

Obesity is believed to be a major causal factor in the development of type 2 diabetes, which is a growing major public health problem worldwide. Diseases or disorders that can develop as a result of untreated diabetes include cardiovascular and peripheral arterial disease, small and large vessel disease, stroke, and some forms of cancer, in particular cancers of the hematopoietic system.

There is a need in the art for additional amylin analogs. For example, amylin analogs that exhibit reduced fibrillation and/or high chemical stability at or near pH 7 may provide a formulation at or near physiological pH. Amylin analogs having plasma half-lives of suitable length may also allow for longer dosing intervals than is currently possible (eg, weekly or even less frequent administration) and hence improve patient compliance. High levels of amylin receptor agonist activity may also be desirable.

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Brief description of the invention

The present invention relates to compounds that are analogues of human amylin.

In a first aspect, the present invention provides an amylin analogue which is a compound having the formula

R1-ZR ² , where

R ¹ is hydrogen, C ₁ _ ₄ acyl, benzoyl or C ₁ _ ₄ alkyl, or a half-life increasing moiety M, where M is optionally linked to Z via a linker moiety L;

R ² is OH or NHR ³ where R ³ is hydrogen or C ₁ _ ₃ alkyl; And

Z is the amino acid sequence of formula I:

X1-X2-X3-X4-X5-X6-X7-Ala-Thr-X10-Arg-Leu-Ala-X14-Phe-Leu-X17-Arg-X19-X20-PheGly(Me)-Ala-lle(Me )-X27-Ser-Ser-Thr-Glu-X32-Gly-Ser-X35-Thr-X37 (I), where

X1 is selected from the group consisting of Arg, Lys and Glu;

X3 is selected from the group consisting of Gly, Gln and Pro;

X4 is selected from the group consisting of Thr and Glu;

X5 is selected from the group consisting of Ala and Leu;

X6 is selected from the group consisting of Thr and Ser;

X10 is selected from the group consisting of Glu and Gln;

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

X17 is selected from the group consisting of Gln, His and Thr;

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

X27 selected from a group consisting of Leu and Pro;

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

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

X37 is selected from the group consisting of Hyp and Pro and

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

or a pharmaceutically acceptable salt or solvate thereof.

In some embodiments, X1 is selected from Arg and Lys.

In some embodiments, X3 is Gly, X4 is Thr, X5 is Ala, and/or X6 is Thr, e.g., X3 is Gly, X4 is Thr, X5 is Ala, and X6 is Thr.

In some embodiments, X14 is selected from His, Asp, and Aad. In some embodiments, X17 is Gln.

In some embodiments, X19-X20 is selected from Ser-Ser and Thr-Thr, or absent, for example, is Ser-Ser.

In some embodiments, X32 is Val, X35 is Asn, and/or X37 is Hyp.

Therefore, Z may be the amino acid sequence of formula II:

X1-X2-Gly-Thr-Ala-Thr-X7-Ala-Thr-X10-Arg-Leu-Ala-X14-Phe-Leu-Gln-Arg-X19-X20-PheGly(Me)-Ala-lle(Me )-X27-Ser-Ser-Thr-Glu-Val-Gly-Ser-Asn-Thr-Hyp (II), where

X1 is selected from the group consisting of Arg and Lys;

X10 is selected from the group consisting of Glu and Gln;

X14 is selected from the group consisting of Aad, Asp and His;

X19-X20 is selected from Ser-Ser and Thr-Thr or absent;

X27 is selected from the group consisting of Leu and Pro and

X2 and X7 are amino acid residues whose side chains together form a lactam bridge.

In some embodiments, X14 is Aad, X19-X20 is SerSer, and X27 is Leu.

In the present specification, the amino acid positions of the amylin analogs are numbered according to the corresponding position in native human amylin having the sequence shown above. The sequences of formulas I and II (and other formulas herein) contain a deletion of two amino acids corresponding to the two residues Asn21 and Asn22 of human amylin.

Thus, for ease of comparison with the amylin sequence, the C-terminal (downstream) Phe residue immediately after position X20 is designated as position 23 because it corresponds to human amylin Phe23. Therefore, the numbering of any single residue in Formulas I and II above and in other formulas throughout this specification indicates the corresponding human amylin residue, provided it is optimally aligned with it, and does not necessarily indicate the linear position of said residue. in a specific sequence.

It is obvious that any of the corresponding formulas presented in the present description, can be represented so as to include residues X21-X22 in the appropriate positions, where X21 and X22 are absent.

It is known that native amylin almost instantly forms fibrils in aqueous solution. Consequently, many attempts have been made to improve the stability of amylin analogues in liquid formulations. The fibrillation propensity can be reduced by introducing N-methylated residues (mentioned above) and/or by changing certain amino acids at various positions. However, despite these possibilities, there remains a need to further optimize the stability of amylin analogues in aqueous solution. Amylin analogues with further improved chemical stability in aqueous solution will facilitate the development of an appropriate pharmaceutical product, possibly even in the form of a ready-to-use formulation, for example, having a pH in the range of neutral or close to it (pH 7-7.4).

Native amylin and the vast majority of amylin analogs (such as pramlintide) contain a disulfide bridge between cysteine residues at positions 2 and 7. The internal cyclization that this bridge provides appears to be necessary for full efficacy and activity. Although compounds containing an internal disulfide bond are often less chemically stable than may be desirable, and the presence of said bond may promote dimerization and oligomerization, for example by disulfide exchange reactions, it has not been reported that the disulfide bond in amylin or amylin analogues is a factor related to low chemical stability in aqueous formulations.

Several attempts have been reported to replace the disulfide bridge in amylin. As described above, As described above, in an attempt to improve in vivo stability and efficacy by reducing chemical and enzymatic proteolysis, WO 91/07978 proposes to replace the internal disulfide bridges of hypocalcemic peptides (including, for example, calcitonin and amylin) with alternative cyclic structures. However, the effect of these alternative structures on the activity of amylin analogs has not been disclosed. WO 99/34764 presents data showing that the replacement of a disulfide bridge with an intramolecular lactam bridge in the sequence of native human amylin results in an amylin analog ( ^2,7 cyclo-[2Asp,7Lys7Lys]-h-amylin) with a significantly lower potency than wild-type amylin and many other amylin analogues, which may further explain why alternative ring structures have not been further pursued.

However, it has now been found that lactam-based ring structures are well compatible with amylin analogs having deletions at positions 21 and 22, since the replacement of the disulfide bridge by the lactam bridge leads to a significant increase in stability in aqueous solution (see Table 2), in while other beneficial properties of these amylin analogues, such as low fibrillation, high activity and good solubility, are retained. Although such lactam bridges are known to drastically reduce the activity of peptides (see WO 91/07978, p. 45, lines 36-52; WO 99/34764, p. 84, table A), in addition to this, currently It has now been found that the amylin analogs of the present invention can retain high activity/show no decrease in activity at the hCT-R, hAMYR1, hAMYR2 and/or hAMYR3 receptors. Additionally or alternatively, they may have excellent chemical and fibrillation stability, particularly, but not exclusively, in the neutral pH range.

Thus, the amylin analog of the present invention contains a lactam bridge formed between the side chains of the residues at positions X2 and X7. For simplicity, positions 2 and 7 will be discussed with reference to residues conditionally present prior to lactam formation.

One of the residues at positions X2 and X7 is a residue with a side chain containing a carboxylic acid group, and the other is a residue with a side chain containing an amine group, where lactam (cyclic amide) is formed between the carboxylic acid and amine groups. The amine may be a primary or secondary amine, but is generally a primary amine. Suitable amino acids whose side chains can participate in the formation of a lactam bridge include Asp, Glu and Aad (having side chains containing carboxylic acid groups) and Dap, Dab, Orn, Lys and hLys (having side chains containing amine groups). Any of the amino acids selected from Asp, Glu and Aad can essentially form a lactam bridge with any of the amino acids selected from the group consisting of Dap, Dab, Orn, Lys and hLys.

Thus, one of the residues at position X2 and X7 may be selected from Asp, Glu and Aad and the other may be selected from Dap, Dab, Orn, Lys and hLys.

In some embodiments, the carboxylic acid moiety of the lactam bridge is derived from the amino acid at position X2, while the amine moiety of the lactam bridge is derived from the amino acid at position X7. Thus, X2 can be selected from Asp, Glu and

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Aad, and X7 may be selected from Dap, Dab, Orn, Lys and hLys.

It may be advantageous for the side chain of the residue at position X2 to be the same length or shorter than the side chain of the residue at position X7. Such combinations of residues may provide advantages, including higher efficiency, than other combinations.

In this context, side chain length is defined by the number of atoms in the linear chain from the first side chain atom (which is linked to the peptide backbone atom, i.e. the alpha carbon of the corresponding residue for most amino acids) to the atom that participates in the formation of the amide bond of the lactam bridge ( i.e., the carbon atom of the carboxylic acid functional group or the nitrogen atom of the amino group), inclusive.

Thus, conventional side chains containing acid and amine groups are assumed to be of the following length:

Asp: 2 atoms

Glu: 3 atoms

Aad: 4 atoms

Dap: 2 atoms

Dab: 3 atoms

Ogp: 4 atoms

Lys: 5 atoms hLys: 6 atoms

In some embodiments, the side chain of the residue at position X2 is shorter than the side chain of the residue at position X7.

Desirably, the length of the lactam bridge provided by the two side chains after the formation of the amide bond (not including any atoms of the peptide backbone) is 4, 5, 6, 7 or 8 atoms, for example 5, 6, 7 or 8 atoms, or 5 , 6 or 7 atoms.

Thus, suitable pairs of residues at the X2 and X7 positions in which the side chain at the X2 position is shorter than the side chain at the X7 position include:

X2 is Asp and X7 is Lys

X2 is Asp and X7 is Orn

X2 is Asp and X7 is Dab

X2 is Asp and X7 is hLys

X2 is Dap and X7 is Aad.

Examples of suitable pairs of residues having the same side chain length include:

X2 is Glu and X7 is Dab

X2 is Dab and X7 is Glu

Other couples that might still be considered include:

X2 is Asp and X7 is Dap

X2 is Aad and X7 is Dap

X2 is Dap and X7 is Asp

X2 is Dab and X7 is Asp

X2 is Orn and X7 is Asp

Pairs of particular interest are:

X2 is Asp and X7 is Lys

X2 is Asp and X7 is Orn.

In some embodiments of the formulas described above:

X1 may be Arg;

X10 may be Glu.

X14 can be selected from Asp and Aad.

X19-X20 may be Ser-Ser

X27 could be Leu

The amylin analogue may have the formula:

R1-ZR ² , where

R ¹ is hydrogen, C ₁ . ₄ -αcyl, benzoyl or C ₁ . _{a 4} -alkyl or half-life increasing moiety M, where M is optionally linked to Z via a linker moiety L;

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R ² is OH or NHR ³ where R ³ is hydrogen or C _1-3 alkyl; And

Z is an amino acid sequence selected from the group consisting of:

RDOGTAT-DapO-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RDOGTAT-DabO-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RD0GTAT-Orn0-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RD0GTAT-Orn0-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-PSSTEVGSNT-Hyp

RD0GTAT-Orn0-ATERLAHFLQRF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RD0GTAT-Orn0-ATERLAHFLHRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNTP

RD0GTAT-Orn0-ATQRLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RD0GTAT-Orn0-ATQRLAHFLQRSSF-Gly(Me)-A-lle(Me)-PSSTEVGSNT-Hyp

RD0GTAT-Orn0-ATERLARFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

EDOGTATKOATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RDOGEATKOATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RDOGTLTKOATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RDOGTASKOATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RDOGTATKOATQRLAHFLQRSSF-Gly(Me)-A-lle(Me)-PSSTEVGSNT-Hyp

RDOGTATKOATQRLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RDOGTATKOATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-PSSTEVGSNT-Hyp

RDOGTATKOATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RDOGTAT-hLysO-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RD0GTAT-Orn0-ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RD0GTAT-Orn0-ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)-PSSTEVGSNT-Hyp

RD0GTAT-Orn0-ATERLA-Aad-FLTRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSST-Hyp

RD0GTAT-Orn0-ATERLA-Aad-FLQRTTF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RD0GTAT-Orn0-ATERLA-Aad-FLQRTTF-Gly(Me)-A-lle(Me)-PSSTEVGSNT-Hyp

RD0GTAT-Orn0-ATERLA-Aad-FLQRATF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RD0GTAT-Orn0-ATERLA-Aad-FLQRAAF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RD0GTAT-Orn0-ATERLA-Aad-FLQRGTF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

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RD0QTAT-Orn0-ATERLA-Aad-FLQRGTF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RDOPTATKOATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

EDOGTATKOATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)-PSSTEVGSNT-Hyp

RDOGTATKOATERLA-Aad-FLQRAAF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RDOGTATKOATERLA-Aad-FLQRGGF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RDOGTATKOATERLA-Aad-FLQRANF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RDOGTATKOATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)-PSSTEVGSNT-Hyp

RDOGTATKOATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RDOGTATKOATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)-LSSTETGSNT-Hyp

EDOGTATKOATERLA-Aad-FLQRSSFGIy(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RDOGTATKOATERLA-Aad-FLQRTTF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

KDOGTATKOATQRLA-Aad-FLQRSSF-Gly(Me)-Alle(Me)-LSSTEVGSNTHyp

RDOGTATKOATQRLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RDOGTATKOATQRLADFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

RDOGTATKOATQRLADFLQRSSF-Gly(Me)-A-lle(Me)-LSSTETGSNT-Hyp

KDOGTATKOATQRLANFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

KDOGTATKOATQRLANFLQRSSF-Gly(Me)-A-lle(Me)-LSSTETGSNT-Hyp

R-DapO-GTATDOATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

R-DabO-GTATDOATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

R-Orn0-GTATD0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

R-DapO-GTAT-AadO-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

R-DabO-GTATEOATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

R-AadO-GTAT-DapO-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp

REOGTAT-DabO-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp or be a pharmaceutically acceptable salt or solvate of such a compound.

The lactam bridge at positions 2 and 7 is indicated by parentheses () following the residues at the indicated positions.

In some embodiments, R ¹ is M or ML- and/or R ² is NH2.

Specific compounds according to the present invention include:

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[19CD]-isoGlu-RD0GTAT-Dap0-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 1) [19CD]-isoGlu-RD0GTAT-Dab0-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 2) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 3) [19CD]-isoGlu-isoGlu-RD0GTAT-Orn0-ATERLAHFLQRSSF-Gly(Me)-Alle(Me)-LSSTEVGSNT-Hyp-NH ₂ (Compound 4) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 5) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLAHFLQRF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 6) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLAHFLHRSSF-Gly(Me)-A4le(Me)LSSTEVGSNTP-NH ₂ (Compound 7) [19CD]-isoGlu-RD0GTAT-Orn0-ATQRLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 8) [19CD]-isoGlu-RD0GTAT-Orn0-ATQRLAHFLQRSSF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 9) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLARFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 10) [19CD]-isoGlu-ED0GTATK0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 11) [19CD]-isoGlu-RD0GEATK0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 12) [19CD]-isoGlu-RD0GTLTK0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 13) [19CD]-isoGlu-RD0GTASK0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 14) [19CD]-isoGlu-RD0GTATK0ATQRLAHFLQRSSF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 15) [19CD]-isoGlu-RD0GTATKQATQRLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 16) [19CD]-isoGlu-RD0GTATK0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 17)

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[19CD]-isoGlu-RD0GTATK0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 18) [19CD]-isoGlu-RD0GTAT-hLys0-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 19) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 20) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 21) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLA-Aad-FLTRSSF-Gly(Me)-A-lle(Me)LSSTEVGSST-Hyp-NH ₂ (Compound 22) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLA-Aad-FLQRTTF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 23) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLA-Aad-FLQRTTF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 24) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLA-Aad-FLQRATF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 25) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLA-Aad-FLQRAAF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 26) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLA-Aad-FLQRGTF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH2 (Compound 27) [19CD]-isoGlu-RD0QTAT-Orn0-ATERLA-Aad-FLQRGTF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 28) [19CD]-isoGlu-RD0PTATK0ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 29) [19CD]-isoGlu-ED0GTATK0ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 30) [19CD]-isoGlu-RD0GTATK0ATERLA-Aad-FLQRAAF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 31) [19CD]-isoGlu-RD0GTATK0ATERLA-Aad-FLQRGGF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 32) [19CD]-isoGlu-RD0GTATK0ATERLA-Aad-FLQRANF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 33) [19CD]-isoGlu-RD0GTATK0ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 34) [19CD]-isoGlu-RD0GTATK0ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 35) [19CD]-isoGlu-RD0GTATK0ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)LSSTETGSNT-Hyp-NH ₂ (Compound 36) [19CD]-isoGlu-ED0GTATK0ATERLA-Aad-FLQRSSFGIy(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 37)

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[19CD]-isoGlu-RD0GTATK0ATERLA-Aad-FLQRTTF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 38) [19CD]-isoGlu-KD0GTATK0ATQRLA-Aad-FLQRSSF-Gly(Me)-Alle(Me)LSSTEVGSNTHyp-NH ₂ (Compound 39) [19CD]-isoGlu-RD0GTATK0ATQRLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 40) [19CD]-isoGlu-RD0GTATK0ATQRLADFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 41) [19CD]-isoGlu-RD0GTATK0ATQRLADFLQRSSF-Gly(Me)-A-lle(Me)LSSTETGSNT-Hyp-NH ₂ (Compound 42) [19CD]-isoGlu-KD0GTATK0ATQRLANFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 43) [19CD]-isoGlu-KD0GTATK0ATQRLANFLQRSSF-Gly(Me)-A-lle(Me)LSSTETGSNT-Hyp-NH ₂ (Compound 44) [19CD]-isoGlu-R-Dap0-GTATD0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 45) [19CD]-isoGlu-R-Dab0-GTATD0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 46) [19CD]-isoGlu-R-Orn0-GTATD0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 47) [19CD]-isoGlu-R-DapO-GTAT-AadO-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 48) [19CD]-isoGlu-R-Dab0-GTATE0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 49) [19CD]-isoGlu-R-AadO-GTAT-DapO-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 50) [19CD]-isoGlu-RE0GTAT-Dab0-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 51)

and their pharmaceutically acceptable salts and solvates.

The present invention also provides a composition containing an amylin analog as described above. Said composition may be a pharmaceutical composition and may contain a pharmaceutically acceptable carrier, excipient or carrier medium.

The present invention also provides a method for synthesizing the amylin analog described above. The method may include the steps of synthesizing the peptide using a solid phase or liquid phase technique and optionally isolating and/or purifying the final product. The method may further include the step of forming an amide bond between the side chains at positions 2 and 7.

The present invention also provides an amylin analog of the present invention for use in a method of therapeutic treatment.

Amylin analogs are useful for, inter alia, reducing food intake, promoting weight loss, and slowing or limiting weight gain. As a result, they may be used to treat a variety of conditions, diseases, or disorders in a subject, including, but not limited to, obesity and various obesity-related conditions, diseases, or disorders such as diabetes (e.g., type 2 diabetes), hypertension, dyslipidemia. , sleep apnea and cardiovascular disease. The subject may be obese accompanied by at least one overweight associated comorbid condition such as diabetes (eg, type 2 diabetes), hypertension, dyslipidaemia, sleep apnea, and cardiovascular disease. It should be understood that amylin analogs may thus be administered to subjects prone to conditions characterized by poor appetite control or other overnutrition, such as binge eating and Prader-Willi syndrome. It will be appreciated that these analogs may be used to treat combinations of the described conditions.

Thus, the present invention provides an amylin analogue of the present invention for use in a method of treating, slowing or limiting weight gain, promoting weight loss and/or reducing overweight. Treatment can be carried out, for example, by controlling appetite, nutrition, food intake, caloric intake and/or energy expenditure.

The present invention also provides an amylin analog of the present invention for use in a method of treating obesity and related diseases, disorders and health conditions, including but not limited to morbid obesity, preoperative obesity, obesity-related inflammation, obese gallbladder disease and obesity-induced sleep apnea and respiratory disorders, cartilage degeneration, osteoarthritis, and reproductive health complications associated with obesity or overweight, such as infertility. The subject may be obese accompanied by at least one overweight associated comorbid condition such as diabetes (eg, type 2 diabetes), hypertension, dyslipidaemia, sleep apnea, and cardiovascular disease.

The present invention also provides an amylin analog of the present invention for use in a method for the prevention or treatment of Alzheimer's disease, diabetes, type 1 diabetes, type 2 diabetes, pre-diabetes, insulin resistance syndrome, glucose intolerance (IGT), pathological conditions associated with elevated blood glucose, metabolic disease, including metabolic syndrome, hyperglycemia, hypertension, atherogenic dyslipidemia, hepatic steatosis (fatty liver; including non-alcoholic fatty liver disease (NAFLD), which in turn includes non-alcoholic steatohepatitis (NASH)), renal failure , arteriosclerosis (eg, atherosclerosis), large vessel disease, small vessel disease, diabetes-induced heart disease (including diabetic cardiomyopathy and heart failure as a complication of diabetes), coronary artery disease, peripheral arterial disease, or stroke, and combinations thereof.

The present invention also provides an amylin analog of the present invention for use in a method for reducing circulating LDL and/or increasing HDL/LDL ratio.

The effect of amylin analogs on these conditions may be wholly or partly mediated by the effect on body weight, or may be independent of it.

The present invention further provides the use of an amylin analog of the present invention for the manufacture of a medicament for treating, slowing or limiting weight gain, promoting weight loss and/or reducing overweight.

The present invention also provides the use of an amylin analog of the present invention for the preparation of a medicament for the treatment of obesity and related diseases, disorders and health disorders, including, but not limited to, morbid obesity, preoperative obesity, obesity-related inflammation, obesity-related gallbladder disease and obesity-induced sleep apnea and respiratory disorders, cartilage degeneration, osteoarthritis, and reproductive health complications associated with obesity or overweight, such as infertility. The subject may be obese accompanied by at least one overweight associated comorbid condition such as diabetes (eg, type 2 diabetes), hypertension, dyslipidaemia, sleep apnea, and cardiovascular disease.

The present invention also provides the use of an amylin analog of the present invention for the preparation of a medicament for the prevention or treatment of Alzheimer's disease, diabetes, type 1 diabetes, type 2 diabetes, prediabetes, insulin resistance syndrome, glucose intolerance (IGT), pathological conditions, associated with elevated blood glucose, metabolic disease, including metabolic syndrome, hyperglycemia, hypertension, atherogenic dyslipidemia, hepatic steatosis (fatty liver; including non-alcoholic fatty liver disease (NAFLD), which in turn includes non-alcoholic steatohepatitis (NASH)), renal failure, arteriosclerosis (eg, atherosclerosis), large vessel disease, small vessel disease, diabetes-induced heart disease (including diabetic cardiomyopathy and heart failure as a complication of diabetes), coronary artery disease, peripheral arterial disease or stroke, and combinations thereof.

The present invention also provides the use of an amylin analogue of the present invention for the preparation of a medicament for reducing circulating LDL and/or increasing the HDL/LDL ratio.

The present invention also provides a method for treating, slowing or limiting weight gain, promoting weight loss and/or reducing overweight in a subject, comprising administering to the subject a therapeutically effective amount of an amylin analog of the present invention.

The present invention also provides a method for the treatment of obesity and related diseases, disorders and health disorders, including, but not limited to, morbid obesity, preoperative obesity, obesity-related inflammation, obesity-related gallbladder disease, and obesity sleep apnea. sleep and respiratory disorders, cartilage degeneration, osteoarthritis, and reproductive health complications associated with obesity or overweight, such as infertility, in a subject, comprising administering to the subject a therapeutically effective amount of an amylin analog of the present invention. The subject may be obese accompanied by at least one overweight associated comorbid condition such as diabetes (eg, type 2 diabetes), hypertension, dyslipidaemia, sleep apnea, and cardiovascular disease.

The present invention also provides a method for preventing or treating Alzheimer's disease, diabetes, type 1 diabetes, type 2 diabetes, prediabetes, insulin resistance syndrome, glucose intolerance (IGT), pathological conditions associated with elevated blood glucose, metabolic disease including metabolic syndrome, hyperglycemia, hypertension, atherogenic dyslipidemia, hepatic steatosis (fatty liver; including non-alcoholic fatty liver disease (NAFLD), which in turn includes non-alcoholic steatohepatitis (NASH)), renal failure, arteriosclerosis (eg, atherosclerosis), large vessel disease, small vessel disease, diabetes-induced heart disease (including diabetic cardiomyopathy and heart failure as a complication of diabetes), coronary artery disease, peripheral arterial disease, or stroke, and combinations thereof in a subject, comprising administering to the subject a therapeutically effective amount of an amylin analog of the present invention.

The present invention also provides a method for reducing circulating LDL and/or increasing an HDL/LDL ratio in a subject, comprising administering to the subject a therapeutically effective amount of an amylin analog of the present invention.

The present invention also provides the use of the amylin analog described above in a method for aesthetic (ie non-therapeutic) weight loss. It should be understood that references to the therapeutic use of amylin analogs and methods involving the administration of amylin analogs may equally be considered to encompass the use and administration of such compositions.

Other aspects and embodiments of the present invention will become apparent from the disclosure below.

Detailed description of the invention

Unless otherwise indicated in the present description, scientific and technical terms in the context of the present description should have the meanings commonly understood by those of ordinary skill in the art. In general, the nomenclature used in the present specification relating to the techniques of chemistry, molecular biology, cell and cancer biology, immunology, microbiology, pharmacology, and protein and nucleic acid chemistry described in this specification is well known and widely used in the art. .

All publications, patents and published patent applications mentioned in this application are expressly incorporated herein by reference. In the event of conflict, the present description, including its specific definitions, shall prevail.

Within the scope of this specification, the word include/comprise, or variations thereof such as includes/comprises or including/comprising, shall mean the inclusion of a specified integer or component, or a specified group of integers or components, but not the exclusion of any other integer or component, or groups of integers or components.

The definite and indefinite forms of the singular include the plural forms, unless the context clearly dictates otherwise.

The term including is used to mean including, but not limited to. The terms including and including but not limited to are used interchangeably.

The terms patient, subject, and individual may be used interchangeably and may refer to both human and non-human animals. Subjects are typically mammals, including humans, non-human primates (including great apes, marmosets, and prehensile-tailed monkeys), livestock (e.g., cattle, pigs), companion animals (e.g., dogs, cats) and rodents (eg mice and rats).

In the context of the present description, the term pharmaceutically acceptable salt is intended to mean a salt that is not harmful to the patient or subject to whom said salt is administered. It may be a salt suitably selected from, for example, acid addition salts and basic salts. Examples of acid addition salts include chlorides, citrates and acetates. Examples of basic salts include salts in which the cation is selected from alkali metal cations such as sodium or potassium ions, alkaline earth metal cations such as calcium or magnesium ions, and substituted ammonium ions such as N(R ¹ )(R ² )(R ³ )(R ⁴ ) ⁺ where R ¹ , R ² , R ³ and R ⁴ independently will generally be hydrogen optionally substituted with C ₁ . ₆ -alkyl or optionally substituted C ₂ _ ₆ -alkenyl. Examples of suitable C ₁ . _{The 6} -alkyl groups include methyl, ethyl, 1-propyl and 2-propyl groups. Examples of possible suitable C2-6alkenyl groups include ethenyl, 1-propenyl and 2-propenyl. Other examples of pharmaceutically acceptable salts are described in Remington's Pharmaceutical Sciences, 17th edition, Alfonso R. Gennaro (Ed.), Mark Publishing Company, Easton, PA, USA, 1985 (and its later editions), Encyclopaedia of Pharmaceutical Technology, 3rd edition , James Swarbrick (Ed.), Informa Healthcare USA (Inc.), NY, USA, 2007 and J. Pharm. sci. 66:2 (1977).

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The term solvate in the context of the present invention refers to a complex of defined stoichiometry formed between a solute (in this case a peptide or a pharmaceutically acceptable salt thereof according to the present invention) and a solvent. The solvent in this context may be, for example, water, ethanol or other pharmaceutically acceptable generally low molecular weight - organic substances, such as, but not limited to, acetic acid or lactic acid. When the solvent in question is water, such a solvate is usually referred to as a hydrate.

The term agonist, as used in the context of the present invention, refers to a substance that activates the type of receptor in question, typically by binding to it (ie acting as a ligand).

Each embodiment of the present invention described herein may be used alone or in combination with one or more other embodiments of the present invention.

Within the scope of this specification, if naturally occurring amino acids are not designated by their full name (e.g., alanine, arginine, etc.), they are designated by their conventional three-letter or single-letter abbreviations (e.g., Ala or A for alanine, Arg or R for arginine). etc.). In the case of some less common or non-naturally occurring amino acids (i.e. amino acids other than the 20 encoded by the standard mammalian genetic code), unless indicated by their full name (e.g. sarcosine, ornithine, etc.), their residues are used commonly used three- or four-digit codes, including Orn (ornithine, i.e. 2,5-diaminopentanoic acid), Aib (α-aminoisobutyric acid), Dab (2,4-diaminobutanoic acid), Dap (2,3-diaminopropanoic acid) , Har (homoarginine), γ-Glu (γ-glutamic acid), Gaba (γ-aminobutanoic acid), β-Ala (i.e. 3-aminopropanoic acid), 8Ado (8-amino-3,6dioxaoctanoic acid).

Unless otherwise indicated, the L-isomer forms of the amino acids in question are mentioned.

Additional abbreviations include the following:

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

Pe(Me): N-methylisoleucine

Aad: 2-aminoadipic acid, e.g. (25)-2-aminoadipic acid [also (25)-2-aminohexanedioic acid], also known as homoglutamic acid

Nur: 4-hydroxyproline, e.g. (25,4^-4-hydroxyproline

[also referred to as (4^-4-hydroxy-b-proline]

Dap: 2,3-diaminopropanoic acid, e.g. (25)-2,3-diaminopropanoic acid

Dab: 2,4-diaminobutanoic acid, e.g. (25)-2,4-diaminobutanoic acid hLys: 2-amino-7-aminoheptanoic acid, also known as homolysin, e.g. (25)-2-amino-7-aminoheptanoic acid

The term "therapeutically effective amount" as used herein in the context of the above described methods of treatment or other therapeutic interventions according to the present invention refers to an amount that is sufficient to treat, ameliorate, alleviate or partially stop the clinical manifestations of a particular disease, disorder or condition that is the object of treatment. or other therapeutic intervention under consideration, such as defined by established clinical outcomes or other biomarkers (established or experimental). The therapeutically significant amount can be empirically determined by one of skill in the art depending on the indication to be treated or prevented and depending on the subject to whom said therapeutically significant amount is administered. For example, one skilled in the art can determine one or more of the clinically relevant biological activity measures described herein, such as plasma fat, blood glucose, or insulin release. A person skilled in the art can determine a clinically significant amount using in vitro or in vivo assays. Other exemplary measures include weight gain, weight loss, and blood pressure changes.

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An amount sufficient to achieve any or all of these effects is defined as a therapeutically effective amount. The amount administered and the route of administration can be adjusted to achieve optimum efficacy. The amount effective for this purpose will depend, among other things, on the severity of the disease, disorder or condition being the object of a particular treatment or other therapeutic intervention, on the body weight and general condition of the subject in question, diet, possible concomitant drug therapy and other factors, well known to medical professionals. Determining the appropriate dose size and administration schedule most suitable for administering to a human a peptide or a pharmaceutically acceptable salt or solvate thereof according to the present invention may be based on the results obtained with the present invention and may be confirmed in suitably planned clinical studies. The effective dose and protocol of treatment can be determined by conventional means, starting with a low dose in laboratory animals and then increasing the dose when the effect is observed, as well as systematically changing the dosage regimen. In determining the optimal dose for a given subject, the attending physician may take into account many factors. Such factors are well known to the skilled person.

The term treatment and its grammatical variations (eg, to be treated, to treat, to treat) as used in the present context refer to an approach to obtain useful or desired clinical results. For the purposes of the present invention, useful or desirable clinical results include, but are not limited to, alleviation of symptoms, reduction in the degree of disease, stabilization (i.e., no worsening) of the course of the disease, delay or slowing of the progression of the disease, improvement or mitigation of the course of the disease, and remission (partial or complete ), both detectable and non-detectable. Treatment can also mean an increase in survival relative to the expected survival time in the absence of treatment. Thus, a subject (eg, human) in need of treatment may be a subject already suffering from the disease or disorder in question. The term treatment includes the suppression or reduction of the increase in the severity of the pathological condition or symptoms (for example, weight gain or hyperglycemia) compared with no treatment and does not necessarily mean the complete cessation of the corresponding disease, disorder or condition.

The term prevention and its grammatical variants (e.g., to be prevented, to prevent, to prevent) as used in the present context refer to an approach to prevent the development of, or prevent the development of, a condition, disease, or disorder, or alteration of the pathology associated with the condition, disease, or disorder. Accordingly, prevention may refer to prophylactic or precautionary measures. For the purposes of the present invention, useful or desirable clinical results include, but are not limited to, preventing or slowing the development of symptoms, progression, or disease progression, whether detectable or not. Thus, the subject (eg, human) in need of prevention may be a subject not yet suffering from the disease or disorder in question. Thus, the term prevention includes the suppression or delay in the onset of a disease compared to no treatment, and does not necessarily mean the long-term prevention of the corresponding disease, disorder or condition.

Synthesis of amylin analogues

The present invention further provides a method for synthesizing an amylin analog of the present invention. Amylin analogs (which may also be referred to as compounds or peptides) can be appropriately prepared using standard synthetic methods. Thus, peptides can be synthesized, for example, using methods comprising synthesizing the peptide using standard solid phase or liquid phase techniques, either stepwise synthesis or fragment assembly, and optionally isolating and purifying the final peptide product. In this context, reference may be made to WO 98/11125 or, inter alia, Fields, G.B. et al., Principles and Practice of Solid-Phase Peptide Synthesis; in: Synthetic Peptides, Gregory A. Grant (ed.), Oxford University Press (2nd edition, 2002) and the synthetic examples provided herein. The method typically further comprises the 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, the cyclization can be carried out in situ on the solid phase (eg, resin), ie before removal of the peptide from the solid phase.

C _1-4 -acyl groups

C ₁ .4-acyl groups that may be present as an R ¹ group in the context of the compounds of the present invention include formyl (i.e., methanoyl), acetyl (i.e., ethanoyl), propanoyl, 1-butanoyl, and 2-methylpropanoyl groups.

C _1-4 -alkyl groups

_C1 . ₄ -alkyl groups which may be present as an R ¹ _{group in the context of the compounds of the present invention include, but are not limited to, C 1-3 alkyl} groups such as methyl, ethyl, 1-propyl, or 2- propyl.

Ci-3-alkyl groups

_C1 . ₃ -alkyl groups that may be present as an R ³ group in the context of the compounds of the present invention include methyl, ethyl, 1-propyl and 2-propyl.

Fragments of M that increase the half-life

As described herein, the N-terminal fragment of R ¹ in the compound of the present invention may be a half-life increasing fragment M (sometimes referred to in the literature as, among other things, a duration-prolonging fragment or an albumin-binding fragment), optionally linked (covalently attached) to a peptide fragment Z via a linker fragment L. Among suitable half-life increasing fragments are certain types of lipophilic substituents. Without wishing to be bound by any particular theory, it is believed that such lipophilic substituents (and other classes of half-life increasing moieties) bind to circulating albumin, thereby protecting the compound of the present invention from renal filtration as well as enzymatic degradation, and thus, probably by increasing the half-life of the compound in vivo. The lipophilic substituent may also modulate the compound's efficacy as an agonist at the amylin (calcitonin) receptor.

The lipophilic substituent may be attached to the N-terminal amino acid residue or to the L linker via an ester, sulfonic acid ester, thioester, amide, amine, or sulfonamide. Accordingly, it should be understood that the lipophilic substituent preferably contains an acyl group, a sulfonyl group, a N atom, an O atom, or an S atom, which are part of an ester, sulfonic acid ester, thioester, amide, amine, or sulfonamide. Preferably the acyl group on the lipophilic substituent is part of an amide or ester with an amino acid residue or a linker.

The lipophilic substituent may contain a hydrocarbon chain containing from 10 to 24 C atoms, for example, from 14 to 22 C atoms, for example, from 16 to 20 C atoms. It preferably contains at least 14 C atoms and preferably contains 20 C atoms or less . For example, a hydrocarbon chain may contain 14, 15, 16, 17, 18, 19, or 20 carbon atoms. Said hydrocarbon chain may be linear or branched and may be saturated or unsaturated. In addition, it may contain a functional group at the end of the hydrocarbon chain, such as a carboxylic acid group, which may or may not be protected during synthesis. From the above discussion it will also be clear that the hydrocarbon chain preferably contains as a substituent a moiety that participates in attachment to the N-terminal amino acid residue of the peptide fragment Z or to the linker L, for example, an acyl group, a sulfonyl group, an N atom, an O atom, or an atom S.

Most preferably, the hydrocarbon chain contains an acyl group as a substituent and, accordingly, said hydrocarbon chain may be part of an alkanoyl group, for example, a dodecanoyl, 2-butyloctanoyl, tetradecanoyl, hexadecanoyl, heptadecanoyl, octadecanoyl, nonadecanoyl, or eicosanoyl group. Examples of functionalized hydrocarbon chains are 15-carboxypentadecanoyl, 17-carboxyheptadecanoyl, and 19-carboxypentadecanoyl.

As indicated above, the lipophilic substituent M may be linked to the N-terminal amino acid residue Z via a linker L. In embodiments, the linker fragment L itself may contain one, two, three or more linked sub-fragments L ¹ , L ² , L ³ , .. etc. In cases where the linker L contains only one such fragment, it is attached to the lipophilic substituent and to the N-terminal amino acid residue of Z. Therefore, the specified linker can be independently attached to the lipophilic substituent and to the N-terminal amino acid residue of Z via an ester, sulfonyl ester, thioether, amide, amine or sulfonamide bond. Accordingly, it may contain two moieties independently selected from acyl, sulfonyl, N atom, O atom, and S atom. The linker may consist of a straight or branched C1-10 hydrocarbon chain, or more preferably a linear _C1-5 hydrocarbon chain. In addition, the linker may contain 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.

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

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References to γ-Glu, ε-Lys and β-Asp indicate amino acid residues that are involved in the formation of bonds through carboxyl or amine functional groups of their side chains. Thus, γ-Glu and β-Asp participate in the formation of bonds through the alpha-amino groups and carboxyl groups of their side chains, while ε-Lys participates through the carboxyl group and the amine group of its side chain. In the context of the present invention, the names γ-Glu and iso-GIu are used interchangeably.

In some embodiments, the linker contains one, two, or three residues independently selected from Glu, γ-Glu, ε-Lys, β-Ala, 4-aminobutanoyl, 8-aminooctanoyl, or 8Ado, or consists of one, two, or three of these leftovers.

In particular, linkers consisting of u3oG1u and u3O-G1u-u3O-G1u may be preferred.

An example of a lipophilic substituent containing a lipophilic fragment M and a linker L is represented by the following formula:

Here, the main chain nitrogen atom of the Arg residue (present at position X1 of the peptide sequence Z of the amylin analog) is covalently attached to the side chain carboxyl group of the Glu moiety via an amide bond. The 19-carboxynodecanoyl group is covalently linked to the alpha-amino group of the Glu linker via an amide bond. Thus, the Glu linker is in the iso-Glu (or γ-Glu) configuration. This combination of a lipophilic moiety and a linker attached to an Arg residue may be denoted by the abbreviation [19CD]-isoGlu-R, for example, when presented in specific compound formulas.

The person skilled in the art is well aware of suitable procedures for the preparation of compounds used in the context of the present invention. For examples of suitable chemical processes see, for example, WO 98/08871, WO 00/55184, WO 00/55119, Madsen et al. (J. Med. Chem. 2007, 50, 6126-32) and Knudsen et al. 2000 (J. Med Chem. 43, 1664-1669).

The hydrocarbon chain in the lipophilic substituent may be further substituted. For example, it may additionally contain up to three substituents selected from NH ₂ , OH and COOH. If the hydrocarbon chain is further substituted, it preferably contains only one substituent. Alternatively or additionally, the hydrocarbon chain may contain a cycloalkane or heterocycloalkane moiety, for example as shown below:

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

In alternative embodiments of the present invention, the N-terminal amino acid Z in the compound of the present invention may be linked (covalently attached) to a biotinyl substituent, optionally via a linker fragment L. Without wishing to be bound by any particular theory, it is believed that such biotinyl substituents bind to albumin in the circulation, thereby protecting the compound of the present invention from enzymatic degradation and thus likely increasing the in vivo half-life of the compound. The linker, if present, can provide a distance between the Z peptide fragment and the biotinyl substituent.

The biotinyl substituent may be attached to an N-terminal amino acid residue or to a linker via a maleimide ester bond, a sulfonyl ester bond, a thioether bond, an amide bond, an amine bond, or a sulfonamide bond. Accordingly, it should be understood that the biotinyl substituent preferably contains a maleimido group, an acyl group, a sulfonyl group, a N atom, an O atom, or an S atom, which participate in the formation of the ester, sulfonyl ester, thioether, amide, amine or sulfonamide bond in question.

Examples of biotinyl substituents may include

Biotin, also known as vitamin H, or coenzyme R, is a water-soluble vitamin.

- 15 042757 group B (vitamin B7). It has been shown to increase the absorption efficiency of some drugs when administered orally.

Connection Efficiency

The compounds of the present invention are amylin receptor agonists, that is, they are capable of binding to one or more receptors or receptor complexes considered to be physiological receptors for human amylin and causing signal transduction involving them. These receptors include the human calcitonin receptor hCT-R as well as complexes containing the human calcitonin receptor hCT-R and at least one of the human receptor activity-modifying proteins referred to as hRAMP1, hRAMP2 and hRAMP3. The complexes between hCT-R and hRAMP1, hRAMP2 and hRAMP3 are designated hAMYR1, hAMYR2 and hAMYR3 (i.e. human amylin receptors 1, 2 and 3), respectively.

Without wishing to be bound by any particular theory, a compound may be considered an amylin receptor agonist if it has agonist activity at one or more of hAMYR1, hAMYR2 and hAMYR3, e.g. hAMYR1 and/or hAMYR3, e.g. hAMYR3.

Typically, an amylin receptor agonist will also have hCT-R agonist activity when expressed in the absence of hRAMP1, hRAMP2 and hRAMP3. Typically, an agonist will have an hCT-R activity (when expressed in the absence of hRAMP1, hRAMP2, and hRAMP3) that is less than 10-fold greater than its activity against any of hAMYR1, hAMYR2, and hAMYR3 (i.e., its activity against all these receptors) in a comparable analysis. hCT-R agonist activity may be less than 5-fold greater than hAMYR1, hAMYR2, and hAMYR3 agonist activity, substantially equal (e.g., +/- 10%) hAMYR1, hAMYR2, and hAMYR3 agonist activity, or be lower agonistic activity against hAMYR1, hAMYR2 and hAMYR3. In this regard, it may be quite simple to compare the activity against hCT-R and hAMYR3.

The ability to induce cAMP production (ie, induce adenylate cyclase activity) by binding to the appropriate receptor or receptor complex is generally considered to be indicative of agonist activity. Other intracellular signaling pathways or events can also be used as measures of amylin receptor agonist activity. These may include calcium release, p-arrestin involvement, receptor internalization, kinase activation or inactivation, lipase activation, inositol phosphate release, diacylglycerol release, or nuclear transcription factor translocation.

An appropriate matching assay format uses cells that express hCT-R and differ only in the expression of hRAMP1, 2, and 3. For example, a baseline cell that does not express any of hCT-R, hRAMP1, hRAMP2, and hRAMP3 can be engineered to produce cells that express (i) hCT-R and (ii) one of hAMYR1, hAMYR2 and hAMYR3 (i.e. hCTR plus one of hRAMP1, hRAMP2 and hRAMP3), eg hAMYR3. The base cells will generally be mammalian cells and may be primate cells. They may be non-human primate cells. Preferably, the host cell does not express any of CT-R, RAMP1, RAMP2, or RAMP3 (whether human or native to the host cell, if said host cell is not a human cell). The basal cells may be fibroblasts. Suitable fibroblast basal cells that are not human fibroblasts include African green monkey COS7 cells that do not express native CT-R or RAMP.

Comparative activity can be determined by any suitable means, such as by determining the values of EC ₅₀ as described below. It will be obvious that the recorded biological parameters should be the same for both types of receptors.

The compounds of the present invention may exhibit a number of properties that provide advantages over human amylin and its existing analogues such as pramlintide, IAPP-GI and the analogues described in WO 2012/168430, WO 2012/168431 and WO 2012/168432. Compared to human amylin or any of these analogs, the compounds of the present invention may, for example, exhibit improved efficacy (eg, improved in vitro activity or efficacy at one or more of the hCT-R, hAMYR1, hAMYR2, or hAMYR3 receptors). Additionally or alternatively, the compounds of the present invention may exhibit improved solubility in aqueous media, in particular at pH values in the range of 4 to 7.5 or within a pH range within this range. In addition, the compounds of the present invention may additionally or alternatively exhibit a reduced tendency to fibril in pharmaceutically relevant aqueous media, in particular at pH values in the range of 4 to 7 or within a pH range within this range. In addition, the compounds of the present invention may additionally or alternatively exhibit improved chemical stability (i.e., reduced tendency to chemical degradation) in aqueous media, in particular at pH values in the range of 4 to 9 or in the pH range

- 16 042757 within this range.

The compounds of the present invention may thus be well suited for preparation in acidic environments (eg pH 4) and in neutral or near neutral environments (eg pH 7 or 7.4). Unlike, for example, pramlintide, which generally exhibits insufficient chemical stability and rapid fibril formation in pharmaceutically relevant aqueous media at neutral pH values, the compounds of the present invention may thus be well suited for co-formulation with, for example, insulin, various insulin analogs and/or other therapeutic (eg, anti-diabetic or anti-obesity) agents that require a neutral or near-neutral pH of the formulation.

In general, it is preferable to use a biological assay that allows the determination of intracellular signaling caused by the binding of a compound to the appropriate receptor, as discussed above. Activation of the calcitonin/amylin receptor by the compounds of the present invention (which behave as receptor agonists) induces the formation of cAMP and the activation of intracellular signaling pathways and events. Thus, cAMP production or any other suitable parameter in suitable cells expressing the receptor can be used to detect agonist activity at said receptor.

Suitable assay formats are known to those skilled in the art and examples are provided below. For example, the assays may use the human calcitonin receptor (hCT-R, eg hCT-R isoform 2) or the hAMYR3 receptor (see examples below). Where precursor sequences are mentioned, it should be understood that the assays may use a mature protein lacking a signal sequence.

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

In some embodiments of the compounds of the present invention, the EC ₅₀ for hCT-R is less than 1.5 nM (eg, 0.001 to 1.5 nM).

In some embodiments of the compounds of the present invention, the EC ₅₀ for hCT-R is less than 0.9 nM (eg, 0.001 to 0.9 nM).

In some embodiments of the compounds of the present invention, the hCT-R EC50 is less than 0.5 nM (eg, 0.001 to 0.5 nM).

In some embodiments of the compounds of the present invention, the hCT-R EC50 is less than 0.3 nM (eg, 0.001 to 0.3 nM).

In some embodiments of the compounds of the present invention, the EC ₅₀ for hCT-R is less than 0.2 nM (eg, 0.001 to 0.2 nM).

The hCT-R EC ₅₀ may be indicative of the effect of a compound on food intake, weight gain and/or weight loss. Compounds with lower EC ₅₀ values for hCT-R may have a more pronounced effect on these parameters.

In some embodiments of the compounds of the present invention, the hAMYR3 EC50 is less than 1.0 nM (eg, 0.001 to 1.0 nM).

In some embodiments of the compounds of the present invention, the hAMYR3 EC50 is less than 0.5 nM (eg, 0.001 to 0.5 nM).

In some embodiments of the compounds of the present invention, the hAMYR3 EC50 is less than 0.4 nM (eg, 0.001 to 0.4 nM).

In some embodiments of the compounds of the present invention, the EC ₅₀ for hAMYR3 is less than 0.3 nM (eg, 0.001 to 0.3 nM).

In some embodiments of the compounds of the present invention, the EC ₅₀ for hAMYR3 is less than 0.2 nM (eg, 0.001 to 0.2 nM).

The EC ₅₀ for hCT-R (when expressed in the absence of hRAMP1, hRAMP2 and hRAMP3) may be less than the EC ₅₀ for any or all of hAMYR1, hAMYR2 and hAMYR3, for example hAMYR3.

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

The _EC50 for hCT-R (when expressed in the absence of hRAMP1, hRAMP2 and hRAMP3) may be less than 5-fold lower than the EC50 for any or all of hAMYR1, hAMYR2 and hAMYR3, e.g. for hAMYR3.

EC ₅₀ in relation to hCT-R (when expressed in the absence of hRAMP1, hRAMP2 and hRAMP3) can be

- 17 042757 is substantially equal to (eg +/- 50%) EC ₅₀ for any or all of hAMYR1, hAMYR2 and hAMYR3, eg hAMYR3.

The EC ₅₀ for hCT-R (when expressed in the absence of hRAMP1, hRAMP2 and hRAMP3) may be higher than the EC ₅₀ for any or all of hAMYR1, hAMYR2 and hAMYR3, eg hAMYR3.

Such assays can be carried out under the conditions described in Example 2 below.

Additionally or alternatively, the compounds of the present invention may exhibit excellent resistance to fibrillation. For example, they may show no detectable fibril formation after 96 hours at pH 4.0 and/or pH 7.0, such as at 40°C, such as under the conditions described in Example 4.

Additionally or alternatively, the compounds of the present invention may exhibit excellent chemical stability, ie resistance to degradation in solution. For example, they may be at least 70% pure, at least 75% pure, at least 80% pure, at least 85% pure, at least 90% pure, or at least 95% pure after incubation at pH. 4, pH 6 and/or pH 7 at 40°C for 72 hours or 14 days, for example under the conditions described in example 5.

Therapeutic use

The compounds of the present invention are suitable for use in, inter alia, reducing food intake, promoting weight loss and slowing or limiting weight gain. Thus, they may provide a promising treatment option for, inter alia, obesity and metabolic diseases caused by, characterized by, or associated with overweight.

Thus, these compounds can be used in a method of treating, slowing or limiting weight gain, promoting weight loss, reducing food intake and/or reducing excess body weight. Treatment can be carried out, for example, by controlling appetite, nutrition, food intake, caloric intake and/or energy expenditure.

These compounds can be used in a method of treating obesity as well as related diseases, disorders and health disorders, including, but not limited to, morbid obesity, obesity before surgery, obesity-related inflammation, obesity-related gallbladder disease and obesity-induced sleep apnea and respiratory disorders, cartilage degeneration, osteoarthritis, and reproductive health complications associated with obesity or overweight, such as infertility.

These compounds can also be used in a method for preventing or treating Alzheimer's disease, diabetes, type 1 diabetes, type 2 diabetes, prediabetes, insulin resistance syndrome, glucose intolerance (IGT), pathological conditions associated with elevated blood glucose, metabolic disease, including metabolic syndrome, hyperglycemia, hypertension, atherogenic dyslipidemia, hepatic steatosis (fatty liver; including non-alcoholic fatty liver disease (NAFLD), which in turn includes non-alcoholic steatohepatitis (NASH)), renal failure, arteriosclerosis (eg, atherosclerosis ), large vessel disease, small vessel disease, diabetes-induced heart disease (including diabetic cardiomyopathy and heart failure as a complication of diabetes), coronary heart disease, peripheral arterial disease, or stroke.

These compounds may also be useful for lowering circulating LDL and/or increasing the HDL/LDL ratio.

The action of the compounds described above may be wholly or partly mediated by the effect on body weight, or may be independent of it.

Metabolic syndrome is characterized by a group of metabolic risk factors in one person. These include abdominal obesity (excess fatty tissue around the internal organs of the abdomen), atherogenic dyslipidemia (blood fat disorders including high triglycerides, low HDL cholesterol, and/or high LDL cholesterol that contribute to plaque formation in the walls of arteries), high blood pressure (hypertension), insulin resistance and impaired glucose tolerance, a prothrombotic state (eg, high levels of fibrinogen or plasminogen activator-1 inhibitor in the blood), and a pro-inflammatory state (eg, increased levels of C-reactive protein in the blood).

Individuals with metabolic syndrome are at increased risk of developing coronary heart disease and other diseases associated with other manifestations of arteriosclerosis (eg, stroke and peripheral vascular disease). The dominant underlying risk factor for this syndrome appears to be abdominal obesity.

Pharmaceutical compositions

Also within the scope of the present invention are compositions such as pharmaceutical compositions containing amylin analogs. As with aspects of the present invention, it is to be understood that reference to an amylin analogue encompasses reference to pharmaceutically acceptable salts and solvates.

The amylin analogs of the present invention may be formulated into pharmaceutical compositions which are suitable for administration with or without storage and which generally contain a therapeutically effective amount of at least one peptide of the present invention together with a pharmaceutically acceptable carrier, excipient. or carrier medium.

The term pharmaceutically acceptable carrier includes any of the standard pharmaceutical carriers. Pharmaceutically acceptable carriers for therapeutic use are well known in the art of pharmacy and are described, for example, in Remington's Pharmaceutical Sciences, 17th edition, Alfonso R. Gennaro (Ed.), Mark Publishing Company, Easton, PA, USA, 1985. For example, there may be Sterile saline and phosphate buffered saline with a slightly acidic or physiological pH are used. Suitable pH buffering agents can be, for example, phosphate, citrate, acetate, tris(hydroxymethyl)aminomethane (TRIS), K-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPS), ammonium bicarbonate, diethanolamine, histidine, arginine, lysine or acetate (eg sodium acetate), or mixtures thereof. The term further encompasses any carrier agents listed in the USP for use in animals, including humans.

The pharmaceutical composition according to the present invention may be in unit dosage form. In this form, the composition is subdivided into unit doses containing appropriate amounts of the active component or components. The unit dosage form may be presented as a packaged preparation, the package containing individual quantities of the drug, for example, packaged tablets, capsules or powders in vials or ampoules. The unit dosage form may also be, for example, a capsule, cachet or tablet, or may be a suitable amount of any of these packaged forms. The unit dosage form may also be presented in an injectable form for single administration, for example, in the form of a syringe pen containing a liquid phase (generally aqueous) composition. The compositions may 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 (in / in), intradermal and transdermal) administration or administration by inhalation. For convenience, the formulations may be presented in unit dosage form and may be prepared using any of the methods well known in the art of pharmaceutical formulations.

Subcutaneous or transdermal routes of administration may, in some instances, be suitable for the peptides of the present invention.

Other implementation options relate to devices, dosage forms and packages used to deliver pharmaceutical compositions according to the present invention. Thus, at least one peptide in a stable or preservative formulation or solution described herein can be administered to a patient according to the present invention by various delivery methods, including by subcutaneous or intramuscular injection, or by transdermal, pulmonary or transmucosal administration, or via an implant, or using an osmotic pump, cartridge, micropump, or other means known to those skilled in the art.

Other implementation options relate to formulations for oral administration and oral administration. Oral formulations may use co-administration of adjuvants (e.g., resorcinols and/or nonionic surfactants such as polyoxyethylene oleyl ether and n-hexadecyl polyethylene ether) to artificially increase intestinal permeability and/or co-administration of enzyme inhibitors (e.g. pancreatic trypsin, diisopropylfluorophosphate (DFF) or trasylol) to inhibit enzymatic degradation. A compound that is an active ingredient in a solid oral dosage form may be mixed with at least one additive such as sucrose, lactose, cellulose, mannitol, trehalose, raffinose, maltitol, dextran, starches, agar, alginates, chitins, chitosans, pectins, gum tragacanth, gum arabic, gelatin, collagen, casein, albumin, synthetic or semi-synthetic polymer or glyceride. These dosage forms may also contain other types of additives, such as an inactive diluent, a lubricant (such as magnesium stearate), a paraben, a preservative (such as sorbic acid, ascorbic acid, or alpha-tocopherol), an antioxidant (such as cysteine), a disintegrant, a binder substance, thickener, buffering agent, pH adjusting agent, sweetening agent, flavoring agent or flavoring agent.

Doses

A typical dose of an amylin analogue used in the context of the present invention may range from about 0.0001 to about 100 mg/kg body weight per day, as from about 0.0005

- 19 042757 to about 50 mg/kg of body weight per day, as from about 0.001 to about 10 mg/kg of body weight per day, for example, from about 0.01 to about 1 mg/kg of body weight per day, administered in one or more doses, as from one to three doses. The exact dose used will depend, among other things, on: the nature and severity of the disease or disorder being treated, the sex, age, body weight and general condition of the subject being treated, the possible other concomitant disease or disorder being or should be being treated , as well as other factors that will be known to the practitioner - a specialist in this field of technology.

The amylin analog of the present invention may be administered continuously (e.g., by intravenous administration or other continuous drug delivery method) or may be administered to a subject at predetermined intervals, typically at regular intervals, depending on the desired dose and pharmaceutical composition selected by the practitioner. for a specific subject. When regularly administered, the intervals between administrations include, for example, administration once a day, twice a day, once every two, three, four, five or six days, once or twice a week, once or twice a month, etc. similar. Such regular peptide dosing regimens under certain circumstances, such as, for example, during long-term dosing, may involve advantageous interruptions for a period of time such that the treated subject reduces or stops taking the drug, often referred to as drug rest. Rest from drugs is suitable, for example, to maintain or restore sensitivity to the drug, in particular during long-term treatment, or to reduce unwanted side effects during long-term treatment of the subject with the drug. The duration of the drug rest depends on the duration of the regular dosing regimen and the purpose of the drug rest (eg, to restore drug sensitivity and/or reduce unwanted side effects with continuous long-term administration). In some embodiments, drug rest may be a reduction in drug dose (eg, to a level below a therapeutically effective amount over a period of time). In other embodiments, drug administration is stopped for a period of time before administration is started again using the same or a different regimen (eg, using a lower or higher dose and/or frequency of administration). Thus, drug rest according to the present invention may be selected from a wide range of times and administration schedules. An exemplary drug rest is two or more days, one or more weeks, or one or more months, up to and including about 24 months of drug rest. For example, a regular daily peptide regimen of the present invention may be interrupted with a drug rest of one week, or two weeks, or four weeks, after which the previous regular regimen (eg, daily or weekly regimen) is resumed. Various other drug rest regimens are contemplated for administration of the peptides of the present invention.

Thus, the peptide can be delivered via an administration schedule that includes two or more administration phases separated by appropriate drug rest phases.

During each administration phase, the peptide is administered to the treated subject in a therapeutically effective amount according to a predetermined administration schedule. Said administration schedule may include continuous administration of the drug to the treated subject during the administration phase. Alternatively, said administration schedule may include administering to a treated subject multiple doses of the peptide, where said doses are separated by intervals between administrations.

The administration schedule may include at least two doses per administration phase, at least five doses per administration phase, at least 10 doses per administration phase, at least 20 doses per administration phase, at least 30 doses per administration phase, or more .

Said intervals between administrations may be intervals to ensure regular administration and may be as defined above, including administration once a day, twice a day, once every two, three, four, five or six days, one or twice a week, once or twice a month, or regularly and even less often, depending on the specific dosage form, bioavailability and pharmacokinetic profile of the peptide.

The administration phase may have a duration of at least two days, at least one week, at least 2 weeks, at least 4 weeks, at least one month, at least 2 months, at least 3 months, at least 6 months or more.

If the administration schedule includes multiple doses, the duration of the possible subsequent drug rest phase exceeds the interval between administrations used in this administration regimen. If the intervals between administrations are unequal, the duration of the drug rest phase may exceed the average interval between doses during the administration phase. Alternatively, the duration of drug rest may exceed the longest interval between

- 20 042757 strictly administered doses during the administration phase.

The duration of the possible drug rest phase may be at least twice as long as the duration of the corresponding interval between injections (or its average value), at least 3 times, at least 4 times, at least 5 times, at least 10 times or at least 20 times greater than the duration of the corresponding interval between injections or its average value.

Within these limitations, the drug rest phase may be at least two days, at least one week, at least 2 weeks, at least 4 weeks, at least one month, at least 2 months, at least 3 months, at least 6 months or more depending on the administration schedule during the preceding administration phase.

The regimen of administration, involving the use of drug rest, includes at least 2 phases of administration. Successive administration phases are separated by corresponding drug rest phases. Thus, an administration schedule may include at least 3, at least 4, at least 5, at least 10, at least 15, at least 20, at least 25, or at least 30 or more administration phases. , each of which is separated by appropriate rest phases from drugs.

In successive phases of administration, the same administration schedule may be used, although this may not always be desirable or necessary. However, if other drugs or active agents are administered in combination with the peptide of the present invention, then generally the same combination of drugs or active agents is administered in successive administration phases. In some embodiments, the subject receiving treatment is a human.

Combination Therapy

The amylin analog of the present invention may be administered as part of a combination therapy with another active agent for the treatment of the disease or disorder in question, for example, an antidiabetic agent, an antiobesity agent, an agent for treating metabolic syndrome, an antidyslipidemic agent, an antihypertensive agent, a proton pump inhibitor, or anti-inflammatory agent. In such cases, the two active agents may be administered together or separately, for example as components of the same pharmaceutical composition or formulation, or as separate formulations.

Thus, the peptide of the present invention may have some advantages when administered in combination with a known type of antidiabetic agent, including, but not limited to, metformin, a sulfonylurea, glinide, a DPP-IV inhibitor, glitazone, a GLP-1 receptor agonist (including GLP- 1 or GLP-1 analog, exendin-4 or exendin-4 analog, any other GLP-1 receptor agonist, including liraglutide (Saxenda™, Victoza™), dulaglutide or albiglutide, or dual glucagon-GLP receptor agonist -1, such as described in WO 2008/101017, WO 2008/152403, WO 2010/070252, WO 2010/070253, WO 2010/070255, WO 2010/070251, WO 2011/006497, WO 2011/160630 160633, WO 2013/092703, WO 2014/041195, WO 2015/055801, WO 2015/055802), an SGLT2 inhibitor (i.e. an inhibitor of sodium-glucose transport, e.g. gliflozin such as empagliflozin, canagliflozin, dapagliflozin or ipragliflozin), an agonist GPR40 (FFAR1/FFA1 agonist, e.g. faziglyph) or insulin or insulin analogue. Examples of suitable insulin analogs include, but are not limited to, Lantus (Lantus™), Novorapid (Novorapid™), Humalog (Humalog™), Novomix (Novomix™), Actrafan HM (Actraphane™ HM), Levemir (Levemir™), Degludec (Degludec™) and Apidra (Apidra™). Other suitable antidiabetic agents in this context include GLP-1 receptor agonists such as exenatide (Byetta™ and Bidureon™)-exendin-4) and Byetta LAR (Byetta LAR™), lixisenatide (Lyxumia™) ) and liraglutide (Victoza™).

In addition, the peptide of the present invention can be used in combination with a known type of anti-obesity agent, including, but not limited to, peptide YY or its analogue, neuropeptide Y (NPY) or its analogue, cannabinoid receptor 1 antagonist, lipase inhibitor, proislet human peptide (HIP), melanocortin 4 receptor agonist, GLP-1 receptor agonist (including GLP-1 or GLP-1 analog, exendin-4 or exendin-4 analog, any other GLP-1 receptor agonist, including liraglutide (Saxenda ™), Victoza™), dulaglutide or albiglutide, or a dual glucagon-GLP-1 receptor agonist, such as those described in WO 2008/101017, WO 2008/152403, WO 2010/070252, WO 2010/070253, WO 2010/ 070255, WO 2010/070251, WO 2011/006497, WO 2011/160630, WO 2011/160633, WO 2013/092703, WO 2014/041195, WO 2015/055801), WO 2015/055801), Orlistat™ (Orlistat™), Orlistat™ (Orlistat™), Orlistat™ (Orlistat™) (Sibutramine™), phentermine, melanin-concentrating hormone 1 receptor antagonist, CCK, amylin, pramlintide, and leptin, and their analogues.

The peptide of the present invention can also be used in combination with a known type of antihypertensive agent, including, but not limited to, an inhibitor

- 21 042757 angiotensin-converting enzyme, angiotensin II receptor blocker, diuretic, beta-blocker or calcium channel blocker.

The peptide of the present invention can also be used in combination with a known type of anti-dyslipidemic agent, including, but not limited to, a statin, fibrate, niacin, a PSCK9 inhibitor or a cholesterol absorption inhibitor.

The peptide of the present invention can also be used in combination with a proton pump inhibitor (i.e., a pharmaceutical agent having pharmacological activity as an H ⁺ /K ⁺ -ATPase inhibitor) of a known type, including, but not limited to, an agent of the benzimidazole derivative type or of the imidazopyridine derivatives such as Omeprazole (Omeprazole™), Lansoprazole (Lansoprazole™) Dexlansoprazole (Dexlansoprazole™), Esomeprazole (Esomeprazole™), Pantoprazole (Pantoprazole™), Rabeprazole (Rabeprazole™), Zolpidem (Zolpidem™), Alpidem (Alpidem™) ), Saripidem (Saripidem™) or Necopidem (Necopidem™).

In addition, with regard to anti-inflammatory treatment, the peptide of the present invention may be advantageous when administered in combination with an anti-inflammatory agent of a known type, including, but not limited to, steroids and corticosteroids such as beclomethasone, methylprednisolone, betamethasone, prednisone, dexamethasone, and hydrocortisone;

non-steroidal anti-inflammatory drugs (NSAIDs), such as propionic acid derivatives (eg, alminoprofen, benoxaprofen, bucloxic acid, carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, ibuprofen, indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin, pyrprofen, pranoprofen, suprofen, thiaprofenic acid and thioxaprofen); acetic acid derivatives (eg, indomethacin, acetamethacin, alkofenac, clidanac, diclofenac, fenclofenac, fenclosic acid, fentiazac, furofenac, ibufenac, isoxepac, oxpinac, sulindac, thiopinac, tolmetin, zidomethacin, and zomepirac); fenamic acid derivatives (eg, flufenamic acid, meclofenamic acid, mefenamic acid, niflumic acid, and tolfenamic acid); biphenylcarboxylic acid derivatives (eg diflunisal and flufenisal); oxicams (eg isoxicam, piroxicam, sudoxicam and tenoxicam); salicylates (eg acetylsalicylic acid and sulfasalazine) and pyrazolones (eg apazone, bezpiperilone, feprazone, mofebutazone, oxyphenbutazone and phenylbutazone);

COX II inhibitors such as rofecoxib and celecoxib;

interferon beta preparations (for example, interferon beta-1a or interferon beta-1b);

and certain other compounds such as 5-aminosalicylic acid and its prodrugs and pharmaceutically acceptable salts.

Metformin has also been shown to have anti-inflammatory properties (see, for example, Haffner et al., Diabetes 54: 1566-1572 (2005)) and thus may also be suitable for use in combination with the compounds (peptides) according to the present invention. invention.

Devices and sets

In some embodiments, the present invention relates to a device containing an amylin analogue or a pharmaceutical composition according to the present invention for delivering said analogue to a subject. With such devices, the amylin analogs can be administered to a patient via a variety of delivery methods, including: intravenous, subcutaneous, intramuscular, or intraperitoneal injection; oral administration; transdermal administration; pulmonary or transmucosal administration; introduction using an implant, osmotic pump, cartridge or micropump; or by other means known to the person skilled in the art.

In some embodiments, the present invention relates to a kit containing an amylin analog of the present invention or a pharmaceutical composition of the present invention. In some embodiments, said kit further comprises packaging and/or instructions for use.

Said device or kit may be suitable for use in the combination therapy described above. Thus, the device or kit may further comprise an additional active agent, such as an antidiabetic agent, an antiobesity agent, an agent for the treatment of metabolic syndrome, an antidyslipidemic agent, an antihypertensive agent, a proton pump inhibitor, or an anti-inflammatory agent as described above, or a pharmaceutical composition containing such active agent.

Examples

The following examples demonstrate some specific embodiments of the present invention. The following examples were performed using standard techniques that are well known and are customary to those skilled in the art, unless otherwise detailed. It should be understood that these examples are for illustrative purposes only and are not definitive terms or scope of the present invention. Therefore, they should in no way be construed as limiting the scope of the present invention.

Abbreviations used in the examples include

Ast: ' acetaminomethyl alloc: allyloxycarbonyl Vos: mpem-butoxycarbonyl BSA: bovine serum albumin cAMP: cyclic adenosine monophosphate COMU™: (1-cyano-2-ethoxy-2-oxoethylideneaminooxy)-dimethylaminomorpholinocarbenium hexafluorophosphate DXM: dichloromethane DIPEA: diisopropylethylamine DMEM: Wednesday Needle modified by Dulbecco DMF: Ν,Ν-dimethylformamide DODT: 3,6-dioxa-1,8-octand ithiol ESI-MC: mass spectrometry with electrospray ionization EUN: ethanol _Et2O : diethyl ether FCS: fetal calf serum fmoc: 9-fluorenylmethoxycarbonyl

- 23 042757

HATU: 2-(7-aza-1 H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate HEPES: H-2-hydroxyethylpiperazine-H'-2-ethanesulfonic acid HPLC: high performance liquid chromatography IBMK: 3-isobutyl-1-methylxanthine MeCN: acetonitrile MS: mass spectrometry NEP: N-ethylpyrrolidone NMP N-methylpyrrolidone OAN: allyl ester FSB: phosphate buffered saline p-ERK: phosphorylated extracellular regulated kinase RP-HPLC: reverse phase high performance liquid chromatography TFA: trifluoroacetic acid TIS: triisopropylsilane Trt: trityl (i.e. triphenylmethyl) v/v: volume/volume wt./vol.: mass/volume

The following examples are provided to illustrate some embodiments of the present invention and are not intended to limit the scope of the present invention.

Determination of physiological parameters

Unless otherwise indicated, whole blood glucose was determined in tail vein blood samples using an enzyme-based Biosen electrode (EKF Diagnostic, Germany). Blood samples were analyzed for the presence of glycated hemoglobin (HbA1c) using a Cobas c111 analyzer (Roche Diagnostics, Mannheim, Germany).

Example 1 Synthesis of Compounds

The following compounds have been synthesized:

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[19CD]-isoGlu-RD0GTAT-Dap0-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 1) [19CD]-isoGlu-RD0GTAT-Dab0-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 2) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 3) [19CD]-isoGlu-isoGlu-RD0GTAT-Orn0-ATERLAHFLQRSSF-Gly(Me)-Alle(Me)-LSSTEVGSNT-Hyp-NH ₂ (Compound 4) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 5) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLAHFLQRF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 6) [19CD]-isoGlu-RD0GTAT-Orn()-ATERLAHFLHRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNTP-NH ₂ (Compound 7) [19CD]-isoGlu-RD0GTAT-Orn0-ATQRLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 8) [19CD]-isoGlu-RD0GTAT-Orn0-ATQRLAHFLQRSSF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 9) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLARFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 10) [19CD]-isoGlu-ED0GTATK0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 11) [19CD]-isoGlu-RD0GEATK0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 12) [19CD]-isoGlu-RD0GTLTK0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 13) [19CD]-isoGlu-RD0GTASK0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 14) [19CD]-isoGlu-RD0GTATK0ATQRLAHFLQRSSF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 15) [19CD]-isoGlu-RD0GTATK()ATQRLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 16)

- 25 042757

[19CD]-isoGlu-RD0GTATK0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 17) [19CD]-isoGlu-RD0GTATK0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 18) [19CD]-isoGlu-RD0GTAT-hLys0-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 19) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 20) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 21) [19CD]-isoGlu-RD()GTAT-Orn()-ATERLA-Aad-FLTRSSF-Gly(Me)-A-lle(Me)LSSTEVGSST-Hyp-NH ₂ (Compound 22) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLA-Aad-FLQRTTF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 23) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLA-Aad-FLQRTTF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 24) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLA-Aad-FLQRATF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 25) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLA-Aad-FLQRAAF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 26) [19CD]-isoGlu-RD0GTAT-Orn0-ATERLA-Aad-FLQRGTF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH2 (Compound 27) [19CD]-isoGlu-RD0QTAT-Orn0-ATERLA-Aad-FLQRGTF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 28) [19CD]-isoGlu-RD0PTATK0ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 29) [19CD]-isoGlu-ED0GTATK0ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 30) [19CD]-isoGlu-RD0GTATK0ATERLA-Aad-FLQRAAF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 31) [19CD]-isoGlu-RD0GTATK0ATERLA-Aad-FLQRGGF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 32) [19CD]-isoGlu-RD0GTATK0ATERLA-Aad-FLQRANF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 33) [19CD]-isoGlu-RD0GTATK0ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)PSSTEVGSNT-Hyp-NH ₂ (Compound 34) [19CD]-isoGlu-RD0GTATK0ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 35) [19CD]-isoGlu-RD0GTATK0ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)LSSTETGSNT-Hyp-NH ₂ (Compound 36)

- 26 042757

[19CD]-isoGlu-ED0GTATK0ATERLA-Aad-FLQRSSFGIy(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 37) [19CD]-isoGlu-RD0GTATK0ATERLA-Aad-FLQRTTF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 38) [19CD]-isoGlu-KD0GTATK0ATQRLA-Aad-FLQRSSF-Gly(Me)-Alle(Me)LSSTEVGSNTHyp-NH ₂ (Compound 39) [19CD]-isoGlu-RD0GTATK0ATQRLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 40) [19CD]-isoGlu-RD0GTATK0ATQRLADFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 41) [19CD]-isoGlu-RD0GTATK0ATQRLADFLQRSSF-Gly(Me)-A-lle(Me)LSSTETGSNT-Hyp-NH ₂ (Compound 42) [19CD]-isoGlu-KD0GTATK0ATQRLANFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 43) [19CD]-isoGlu-KD0GTATK0ATQRLANFLQRSSF-Gly(Me)-A-lle(Me)LSSTETGSNT-Hyp-NH ₂ (Compound 44) [19CD]-isoGlu-R-Dap0-GTATD0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 45) [19CD]-isoGlu-R-Dab0-GTATD0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 46) [19CD]-isoGlu-R-Orn0-GTATD0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 47) [19CD]-isoGlu-R-DapO-GTAT-AadO-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 48) [19CD]-isoGlu-R-Dab0-GTATE0ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 49) [19CD]-isoGlu-R-AadO-GTAT-DapO-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 50) [19CD]-isoGlu-RE0GTAT-Dab0-ATERLAHFLQRSSF-Gly(Me)-A-lle(Me)LSSTEVGSNT-Hyp-NH ₂ (Compound 51)

For comparison purposes, three compounds containing disulfide bridges (instead of lactam bridges) and two non-cyclized compounds were synthesized:

[19CD]-isoGlu-RDGTAT-Om-ATERLAHFLQRSSF-Sar-A-lle(Me)-PSSTEVGSNT- Cf. 1

Hyp- _NH2

[19CD]-isoGlu-RC0GTATC0ATERLAHFLQRSSF-Sar-A-lle(Me)-LSSTEVGSNT- Cf. 2

Hyp- _NH2

[19CD]dsoGlu-RDGTAT-Orn-ATERLA-Aad-FLQRSSF-Sar-A4le(Me)- Cf. 3

LSSTEVGSNT-Hyp-NH ₂ [19CD]-isoGlu-RC0GTATC0ATERLA-Aad-FLQRSSF-Sar-A-lle(Me)- Comp. 4

LSSTEVGSNT-Hyp-NH ₂

[19CD]-isoGlu-RC0NTATC0ATQRLADFLQRSSF-Sar-A4le(Me)-LSSTEVGSNT- Cf. 5

Hyp- _NH2

An additional reference compound, designated NN96, is {N-a-[(S)-4carboxy-4-(19-carboxynodecanoylamino)butyryl]-[Arg1,Glu14,His17,Pro37]-pramlintide; disclosed as Example 96 in WO 2012/168430 A2 and has the amino acid sequence RC()NTATC()ATQRLAEFLHHSSNNFGPILPPTNVGSNTP.

Parentheses () indicate intramolecular lactam bridges (or disulfide bridges, if applicable) formed between the side chains of residues at positions 2 and 7 of the respective amino acid sequences.

Unless otherwise noted, the reagents and solvents used in the following examples were commercially available in standard laboratory chemical or analytical grades and were used without further purification.

General Methods for Solid Phase Peptide Synthesis

A Liberty CEM peptide synthesizer or a Liberty Blue CEM peptide synthesizer was used using standard Fmoc reagents. TentaGel™ S Ram resin (1 g; 0.25 mmol/g) was allowed to swell in DMF (10 mL), transferred between tube and reaction vessel with DCM and DMF, and then used. Pseudoprolines [that is, dipeptides used to minimize aggregation during peptide synthesis, such as Fmoc-Phe-Thr(ψ-Me,Me-Pro)-OH, and Fmoc-Asp-Ser(ψ-Me,MePro)-OH, and Fmoc-Ser-Ser(ψ-Me,Me-Pro)-OH] were used where appropriate, and non-naturally occurring amino acids and other suitable building blocks were used without any change to the general procedure.

In the synthesis of compounds, the following optical isomers of specific amino acids were used

- 27 042757 (including non-naturally occurring amino acids):

Hyp: (2S,4R)-4-hydroxyproline [also referred to as (4R)-4-hydroxy-L-proline].

Aad: (25)-2-aminoadipic acid

Dab: (25)-2,4-diaminobutanoic acid

Dap: (25)-2,3-diaminopropanoic acid hLys: (25)-2-amino-7-aminoheptanoic acid, also known as homolysin

Gly(Me): N-methylglycine, also known as sarcosine

Pe(Me): N-methylisoleucine

Coupling reaction:

Liberty CEM Peptide Synthesizer: Fmocamino acid in DMF/DCM (2:1; 0.2 M; 5 mL) was added to the resin in a CEM Discover Microwave Unit along with COMU/DMF (0.5 M; 2 mL) and DIPEA/NMP ( 2.0 M; 1 ml). The coupling reaction mixture was heated to 75° C. for 5 minutes while nitrogen was bubbled through the mixture. The resin was then washed with DMF (4x10 ml). Alternatively, the coupling reaction was carried out without heating, in which case the reaction time was increased to 60 minutes.

Liberty Blue CEM Peptide Synthesizer: Fmoc-amino acid in DMF (0.2 M; 5 mL) was added to the resin in a CEM Discover microwave unit along with DIC/DMF (0.5 M; 2 mL) and Oxyma/DMF (2.0 M; 1 ml). The coupling reaction mixture was heated to 90° C. for 2 minutes while nitrogen was bubbled through the mixture. The resin was then washed with DMF (4x10 ml). Alternatively, the coupling reaction was carried out without heating, in which case the reaction time was increased to 60 minutes.

Regardless of the type of CEM synthesizer, in the case of hindered coupling reactions (eg, addition of a residue immediately after an N-methylated amino acid residue or other sterically hindered amino acid residue known to one skilled in the art), the coupling reaction was repeated one or more times.

Removal of protection:

For initial deprotection, piperidine/DMF (1:4, i.e. 1 part piperidine to 4 parts DMF by volume; 10 ml) was added to the resin and the resulting mixture was heated with microwave radiation (40° C.; 30 sec). The reaction vessel was emptied, a second portion of piperidine/DMF (1:4; 10 ml) was added and reheated (75°C; 3 min). The resin was then washed with DMF (6x10 ml).

Oxidative cyclization

An intramolecular ring formation reaction (disulfide bridge formation) between the Cys residues at positions 2 and 7 (originally attached in the form of protected Acm cysteines) was performed on the peptide still attached to the resin using 163 mg of thallium(III) trifluoroacetate [TI(TFA) ₃ ] in 5 ml NMP at the stage of simultaneous deprotection of Acm and formation of disulfide (alternative method: addition of 10 eq. of iodine to a 50 mM solution of resin-bound peptide in acetic acid and stirring for 18-24 h).

Lactam cyclization

The following Asp-Lys coupling reaction procedure is typical for the preparation of all lactams, wherein the carboxyl-functional amino acid side chain is protected by OAll and the amino group-containing amino acid side chain is protected by Alloc. After assembly of the complete peptide sequence, deprotection of Asp(OAll) at position 2 and Lys(Alloc) at position 7 was performed using 29 mg tetrakis(triphenylphosphine)palladium(0) and 125 μl phenylsilane in 20 ml DCM. A lactam bridge formation reaction between Asp(2) and Lys(7) was then carried out using 414 mg HCTU and 346 μl DIPEA in 20 ml DMF. Both steps were carried out with the peptide still attached to the resin.

Split:

The resin was washed with EtOH (3x10 ml) and Et ₂ O (3x10 ml) and dried at room temperature (RT) to constant weight. The crude peptide was cleaved from the resin by treatment with TFA/TIS/H ₂ O (90:5:5; 40 ml; 2 h; rt) or, alternatively, TFA/DODT (95:5; 40 ml; 2 h; k.t.). Most of the TFA was removed under reduced pressure, the crude peptide was precipitated and washed three times with Et2O and dried at room temperature to constant weight.

Purification and characterization:

The crude peptide was purified by reverse phase preparative HPLC using a gradient of solvent A (0.1% aqueous TFA) and solvent B (0.1% TFA, 90% MeCN in water) using a PerSeptive Biosystems VISION Workstation or a Gilson system system (pumps: Pump 305, 331 Pump, 332 Pump, 402 Syringe Pump; Valvemate® II column switch; UV/Vis-155 UV detector and GX 281 fraction collector; equipped with suitable column and fraction collector) or Waters Autopurification HPLC/MS systems System (pumping system 2525, Waters 2996

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DAD, Waters 2767 sampler, MS ZQ with single quad. Speakers: XSelect CSH 130 Prep

C18 5 mm ODB 30x150 mm or Kinetex 5 mm C8 100A 150x21.2 mm). Fractions were analyzed by analytical HPLC and MS, and the respective fractions were pooled and lyophilized. The final product was characterized by HPLC and MS.

One skilled in the art will appreciate that standard peptide synthesis techniques can be used to prepare the compounds of the present invention.

Example 2 Analysis using hCT-R and hAMYR3

To evaluate the activity of the test peptides in vitro, cell lines expressing the recombinant human calcitonin receptor (hCT-R) or the recombinant human amylin receptor 3 (hAMYR3) from the human astrocytoma cell line 1321N1 were purchased from DiscoveRx Corporation (catalog number 95-0161C6 and 95 -0166C6). hAMYR3 is a heterooligomer of calcitonin receptor (isoform 2; gene ID 799) and RAMP3 (gene ID 10268) that is formed when both genes are expressed in the same cell. The hCT-R cell line expresses only the recombinant human calcitonin receptor gene (isoform 2; gene ID 799). Upon activation of hCT-R or hAMYR3 by test peptides, cAMP production is induced and is determined using the AlphaScreen® cAMP Assay from Perkin-Elmer (catalog number 6760635R).

Briefly, 1321N1 cells expressing hCT-R or hAMYR3 were seeded in 384-well microtiter plates (Falcon Optilux White, catalog number 10448642) at 5000 cells in 50 µl of growth medium per well (AssayComplete 1321N1 Cell Culture Kit, DiscoveRx Corp .) and incubated for 24 h at 37°C, 5% CO2. On the day of analysis, the growth medium was removed and the cells were stimulated by adding 10 μl of stimulation buffer (10 nM Hepes, pH 7.4, 140 nM NaCl, 3.6 nM KCl, 0.5 nM NaH ₂ PO ₄ , 0.5 nM MgSO ₄ , 1.5 nM CaCl ₂ , 5 nM NaHCO ₃ , 0.5 nM IBMC, 0.1% BSA) containing increasing concentrations of the studied peptides, and incubation for 45 min at room temperature. Stimulation was terminated by adding 5.6 µl/well of a detection mixture with donor beads (5 mM Hepes pH7.4, 0.5% TWEEN 20, 0.1% BSA, 0.05 mg/ml donor beads, 62.5 nM biotinylated cAMP) and 4.5 µl/well acceptor bead solution (5 mM Hepes, pH7.4, 0.5% TWEEN 20, 0.1% BSA, 0.05 mg/ml acceptor beads). After thorough mixing, the plates were incubated in the dark for 1 hour at room temperature and the cAMP content in the resulting cell lysates was assessed according to the AlphaScreen® cAMP Assay manufacturer's instructions. EC50 values were evaluated by computer curve fitting of the results for at least 7 different compound concentrations.

The results of the determination of activity in vitro (expressed as values of EC ₅₀ ) are presented below in table. 1.

Example 3 Solubility Determination

The peptide of interest is weighed into a suitable vial and the appropriate buffer (acetate buffer, pH 4.0, phosphate buffer, pH 6, histidine buffer, pH 6 and 7; all at 40 nM) is added to a total volume of 0.5 ml.

The vials are shaken at room temperature for 2 hours and filtered through a filter with a pore diameter of 0.45 μm. The resulting solutions are analyzed by RP-HPLC on a C18 column with gradient elution using formic acid/acetonitrile/water as the mobile phase. The main peak area is determined by UV spectroscopy at 230 nm at each sampling time. The solute concentration is calculated using the external standard method.

Example 4 Assessment of Physical Stability

Aggregation in the form of fibril formation has been detected using the amyloid-specific dye thioflavin T (ThT), which is often used to demonstrate the presence of fibrils in solution (see, for example, Groenning, M., J. Chem. Biol. 3(1 ) (2010), pp. 1-18; Groenning et al., J. Struct. Biol. 158 (2007) pp. 358-369; and Levine, H., III, Protein Sci. 2 (1993) pp. 404 -410). The test peptides (2 mg/ml) were dissolved in demineralized water adjusted to pH 2.5 with HCl at ambient temperature (typically 25°C). Solutions containing (i) 1 mg/ml test peptide, 40 µM ThT and 50 mM phosphate (Ph) buffer (pH 7.0), (ii) 1 mg/ml test peptide, 40 µM ThT and 50 mM histidine (His ) buffer (pH 7.0) and (iii) 1 mg/ml of the test peptide, 40 μM ThT and 50 mM acetate (Ac) buffer (pH 4.0), were placed in 96-well black fluorescent plates (with transparent bottom) in triplicate. Data collection was carried out at fixed intervals of 10 min, each of which was preceded by automatic stirring (shaking) for 300 s, for 96 h at 40°C. Physical stability is determined by measuring the intensity of fluorescence as a function of time. A significant increase in intensity is evaluated as the detection of fibrillation (FF). The data are presented below in table. 1.

Example 5 Evaluation of Chemical Stability

Samples of each test peptide were dissolved in acetate buffer pH 4 and phosphate buffer pH 6 and 7 (all buffers are 40 mM). The final peptide concentration was 1

- 29042757 mg/ml. The samples were placed in glass vials and incubated at 40°C. Samples were analyzed by RP-HPLC on a C8 column with gradient elution using a trifluoroacetic acid/acetonitrile/water system as the mobile phase. The area percentage (area-%) of the main peak was determined using spectroscopy in the UV region at a wavelength of 220 nm at each time point of sampling.

The % degradation was calculated by subtracting the percentage of main peak area at the beginning (t=0) from the percentage of main peak area at each sampling time.

The results of the evaluation of chemical stability after 3 days of incubation are presented as % degradation in table. 1 (below).

The results of the evaluation of chemical stability after 14 days of incubation are presented as % degradation in table. 2 (below).

In an alternative assay, the peptides were dissolved in MilliQ water and adjusted to pH 4.0, 7.5, or 9.0. The samples were placed in glass vials and incubated at 40°C. Samples were analyzed by RP-HPLC on a C8 column with gradient elution using a trifluoroacetic acid/acetonitrile/water system as the mobile phase. The area percentage (area-%) of the main peak was determined using spectroscopy in the UV region at a wavelength of 220 nm at each time point of sampling.

The % degradation was calculated by subtracting the percentage of main peak area at the beginning (t=0) from the percentage of main peak area at each sampling time.

The results of the evaluation of chemical stability after 8 days of incubation are presented as % degradation in table. 3 (below).

Table 1. EC50, chemical stability and fibrillation data

connection no. hCT-R hAMYR3 Fibrillation* (in buffer) % degradation after 72 h at 40°C EC50 [nM] EC50 [nM] pH 4 acetate pH 7 histidine pH 7 Phosphate pH 4 acetate pH 6 phosphate pH 7 Phosphate 1 13.6 9.3 2 0.105 0.032 3 0.081 0.035 NOF NOF OF < 1 3.2 1.2 4 0.160 0.092 NOF NOF NOF < 1 < 1 < 1 5 0.114 0.032 6 0.327 0.011 7 0.289 0.032 8 0.129 0.059 NOF NOF NOF < 1 < 1 < 1

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9 0.027 0.013 10 0.345 0.068 OF NOF NOF < 1 < 1 < 1 eleven 0.068 0.024 NOF NOF NOF < 1 < 1 < 1 12 0.893 0.999 13 1.411 0.456 14 0.386 0.108 15 0.056 0.017 NOF OF OF < 1 < 1 < 1 16 0.043 0.035 17 0.058 0.025 NOF NOF NOF 1.2 < 1 < 1 18 0.085 0.031 NOF NOF NOF < 1 < 1 < 1 19 0.028 0.006 20 0.009 0.003 OF NOF NOF 1.0 1.3 2.0 21 0.037 0.010 NOF NOF NOF < 1 < 1 < 1 22 0.474 0.051 23 1.931 0.706 24 0.038 0.013 25 0.844 0.336 NOF NOF NOF - - < 1 26 0.769 0.120 27 0.521 0.325 NOF NOF NOF < 1 < 1 1.7 28 1.229 0.268 29 1.070 0.312 thirty 0.232 0.122 OF NOF NOF 31 0.227 0.066 32 0.006 0.005 OF NOF NOF 1.3 <1 2.2 33 0.019 0.009 NOF NOF NOF < 1 1 < 1 34 0.206 0.063 NOF NOF NOF < 1 < 1 2.8 35 0.066 0.028 NOF NOF NOF < 1 < 1 1.4 36 0.026 0.007 OF NOF NOF 2.5 2.1 1.0 37 0.116 0.031 OF NOF NOF - < 1 3.0 38 0.114 0.060 OF NOF NOF < 1 < 1 < 1 39 0.094 0.024 OF NOF NOF < 1 < 1 < 1

- 31 042757

40 0.028 0.008 OF NOF NOF 1.1 < 1 < 1 41 0.095 0.024 NOF NOF NOF < 1 < 1 < 1 42 0.006 0.003 43 0.021 0.007 OF OF OF 44 0.020 0.009 OF NOF NOF 45 299 279 46 7.5 5.5 47 10.8 7.7 48 0.107 0.020 49 1.038 0.442 50 3.3 1.5 51 0.861 0.306 Wed 1 20.3 17.1 Wed 2 0.019 0.003 Wed 3 18.2 13.2 Wed 4 0.075 0.031 NOF NOF NOF Wed 5 0.063 0.028 OF NOF NOF

NOF = no fibrillation detected;

OF = fibrillation detected

Table 2. Chemical stability

connection no. % degradation after 14 days at 40°C pH 4 acetate pH 6 phosphate pH 7 Phosphate 32 <1 <1 <1 41 1.1 <1 <1 10 1.9 1.8 9.1 14 2.2 <1 2.6 18 <1 1.0 1.9 37 But <1 <1 36 But 1.6 <1 4 1.6 <1 6.4 38 But But <1 20 But <1 3.7 16 But <1 But 3 But 1.0 But 35 2 <1 <1 Wed 2 <1 4.6 But Wed 4 3.2 12.6 20.3 Wed 5 3.0 5.1 12.7 NN96 3.0 10.1 33.2

n/a - not defined

- 32 042757

Table 3. Chemical stability

connection no. % degradation after 8 days at 40°C pH 4 pH 7.5 pH 9 51 3.1 4.6 4.6 49 1.7 2.1 3.0 48 1.8 2.3 2.5 19 3.9 6.2 4.2

Example 6 Determination of Pharmacokinetic (PK) Profiles in Rats

Male Sprague-Dawley rats were given a single subcutaneous (SC) bolus injection with each of the test peptides listed below.

Compound doses of 30 nmol/kg were administered. Prior to administration and 24 and 96 hours after administration, blood samples were obtained from the tail vein. Immediately after the last blood sampling, the rats were euthanized by shock and cervical dislocation.

The injection medium used for each test peptide was mannitol-containing histidine buffer (pH 7.0). After ethanol precipitation, plasma samples were analyzed by liquid chromatography/mass spectrometry (LC-MS/MS). Mean plasma concentrations were used to calculate pharmacokinetic parameters.

The plasma half-life in the final phase (ti/ ₂ ) was determined by the formula Ιη(2)/λζ, where λζ is the magnitude of the slope of the log-linear regression of the logarithm of concentration versus time during the final phase.

results

The plasma half-lives in the final phase (C/ ₂ ) for all studied peptides were determined in the range of 21.3 to 36.1 hours.

Table 4. In vivo half-life

connection no. tl/2 [h] 3 21.3 4 31.9 18 30.1 20 31.1 35 36.1 38 31.8

Example 7 Effects on food intake and body weight in normal Sprague-Dawley rats in the short term

Sprague-Dawley (SD) rats were obtained from Janvier Labs, France. Animals arrived at least 14 days before the start of the study to adapt to the conditions of the experiment. From arrival and throughout the study, rats were kept in groups of 2 to 4 (n=2-4) in a room with controlled light, temperature and humidity. Animals had ad libitum access to food (KLIBA 3430, Provimi Kliba AG, Switzerland) and water (domestic quality tap water) during the entire study. The group included 6-8 rats. A vehicle treated group and a positive control group were included in each test set. The test peptide was administered to rats subcutaneously (s.c.) once in the morning 1 hour before the lights were turned off, using a weight-adjusted dose (30 nmol/kg). The injection volume was 2 ml/kg. Food intake was recorded in real time using an automatic food recording system (HM02, MBRose, Denmark) for 4 days or manually at 0 h before administration and then 24, 48, 72 and 96 h after administration. Body weight was determined daily.

Statistical analysis was performed using GraphPad™ Prism version 7. Comparison of the determined parameters was performed using two-way ANOVA followed by Dunnett's multiple comparison tests. Differences were considered statistically significant at p < 0.05.

results

At 48 hours post-dose, each of the test compounds (with the exception of compounds 30 and 37) produced a clear, statistically significant reduction in food intake (adjusted for vehicle, in %). This decrease in food intake was reflected in a decrease in body weight (adjusted for the vehicle, in %). Subsequently, normal eating behavior was restored.

Claims (11)

Table 5. Effect on food intake and body weight Compound No. % reduction in food intake after 48 hours Dose: 30 nmol/kg Weight loss in % after 48 hours Dose: 30 nmol/kg 41 75 eleven 35 72 12 18 58 10 39 76 9.2 40 83 7.3 37 24 4.3 thirty < 20 0 36 44 (at a dose of 10 nmol/kg) 5.2 (at a dose of 10 nmol/kg) 4 34 4.6 38 43 8.2 20 49 7.8 15 71 7.7 16 66 7.4 3 34 5.6 Example 8 Post-meal basal glucose changes in diabetic ZDF rats in the short term Male Zucker Diabetic Fatty rats (ZDF-Lepr ^fa /Crl) were obtained from Charles River, USA. Animals adapt to the conditions of the experiment for at least 14 days prior to the start of the study. From the moment of arrival and throughout the study, rats are kept in groups of 2 (n=2) in a room with controlled light, temperature and humidity. Animals have ad libitum access to food (KLIBA 2437, Provimi Kliba AG, Switzerland) and water (domestic quality tap water) during the entire study. Rats are randomly assigned based on blood glucose, HbA1c, and body weight to a specific study group and a vehicle group. The total number of animals per group is 10 (n=10). The test peptide is administered subcutaneously (SC) to rats once in the morning using a weight-adjusted dose (10 nmol/kg). The volume of administration is 2 ml/kg. The glucose content in blood samples taken from the tail vein at 24, 72, 96 and 168 hours after administration, and the blood glucose content was determined using a glucometer (GlucoSmart Swing™; MSP Bodmann GmbH, Germany). Statistical analysis was performed using GraphPad™ Prism version 7. Comparison of the determined parameters was carried out using two-way ANOVA followed by Sidak's criteria for multiple comparisons. Differences are considered statistically significant at p < 0.05. CLAIM 1. Amylin analogue having the formula: [19CD]-isoGlu-RD()GTAT-Dab()-ATERLAHFLQRSSF-Gly(Me)-A- (Compound 2) lle(Me)-LSSTEVGSNT-Hyp-NH2 [19CD]-isoGlu-RD()GTAT-Orn()-ATERLAHFLQRSSF-Gly(Me)-A- (Compound 3) lle(Me)-LSSTEVGSNT-Hyp-NH2 [19CD]-isoGlu-isoGlu-RD()GTAT-Orn()-ATERLAHFLQRSSF- (Compound 4) Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp-NH ₂ [19CD]-isoGlu-RD()GTAT-Orn()-ATERLAHFLQRSSF-Gly(Me)-A- (Compound 5) lle(Me)-PSSTEVGSNT-Hyp-NH2 [19CD]-isoGlu-RD()GTAT-Orn()-ATERLAHFLQRF-Gly(Me)-A- (Compound 6) lle(Me)-LSSTEVGSNT-Hyp-NH2 [19CD]-isoGlu-RD()GTAT-Orn()-ATERLAHFLHRSSF-Gly(Me)-A- (Compound 7) lle(Me)-LSSTEVGSNTP-NH2 [19CD]-isoGlu-RD()GTAT-Orn()-ATQRLAHFLQRSSF-Gly(Me)-A- (Compound 8) lle(Me)-LSSTEVGSNT-Hyp-NH2 [19CD]-isoGlu-RD()GTAT-Orn()-ATQRLAHFLQRSSF-Gly(Me)-A- (Compound 9) lle(Me)-PSSTEVGSNT-Hyp-NH2 [19CD]-isoGlu-RD()GTAT-Orn()-ATERLARFLQRSSF-Gly(Me)-A- (Compound 10) lle(Me)-LSSTEVGSNT-Hyp-NH2 - 34 042757 [19CD]-isoGlu-ED()GTATK()ATERLAHFLQRSSF-Gly(Me)-A- (Compound 11) lie(Me)-LSSTEVGSNT-Hyp-NH ₂ [ 19CD]-isoGiu-RD()GEATK()ATERLAHFLQRSSF-Giy(Me)-A- (Compound 12) lle(Me)-LSSTEVGSNT-Hyp-NH ₂ [ 19CD]-isoGlu-RD()GTASK()ATERLAHFLQRSSF-Gly(Me)-A- (Compound 14) lle(Me)-LSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-RD()GTATK()ATQRLAHFLQRSSF-Gly(Me)-A- (Compound 15) lle(Me)-PSSTEVGSNT-Hyp-NH ₂ [ 19CD]-isoGlu-RD()GTATK()ATQRLAHFLQRSSF-Gly(Me)-A- (Compound 16) lle(Me)-LSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-RD()GTATK()ATERLAHFLQRSSF-Gly(Me)-A- (Compound 17) lle(Me)-PSSTEVGSNT-Hyp-NH2 [19CD]-isoGlu-RD()GTATK()ATERLAHFLQRSSF-Gly(Me)-A- (Compound 18) lle(Me)-LSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-RD()GTAT-hLys()-ATERLAHFLQRSSF-Gly(Me)-A- (Compound 19) lle(Me)-LSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-RD()GTAT-Om()-ATERLA-Aad-FLQRSSF-Gly(Me)- (Compound 20) A-lle(Me)-LSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-RD()GTAT-Om()-ATERLA-Aad-FLQRSSF-Gly(Me)- (Compound 21) A-lle(Me)-PSSTEVGSNT-Hyp-NH2 [19CD]-isoGlu-RD()GTAT-Om()-ATERLA-Aad-FLTRSSF-Gly(Me)- (Compound 22) A-lle(Me)-LSSTEVGSST-Hyp-NH2 [ 19CD]-isoGlu-RD()GTAT-Om()-ATERLA-Aad-FLQRTTF-Gly(Me)- (Compound 24) A-lle(Me)-PSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-RD()GTAT-Om()-ATERLA-Aad-FLQRATF-Gly(Me)- (Compound 25) A-lle(Me)-LSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-RD()GTAT-Om()-ATERLA-Aad-FLQRAAF-Gly(Me)- (Compound 26) A-lle(Me)-LSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-RD()GTAT-Om()-ATERLA-Aad-FLQRGTF-Gly(Me)- (Compound 27) A-lle(Me)-LSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-ED()GTATK()ATERLA-Aad-FLQRSSF-Gly(Me)-A- (Compound 30) lle(Me)-PSSTEVGSNT-Hyp-NH2 [19CD]-isoGlu-RD()GTATK()ATERLA-Aad-FLQRAAF-Gly(Me)-A- (Compound 31) lle(Me)-LSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-RD()GTATK()ATERLA-Aad-FLQRGGF-Gly(Me)-A- (Compound 32) lle(Me)-LSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-RD()GTATK()ATERLA-Aad-FLQRANF-Gly(Me)-A- (Compound 33) lle(Me)-LSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-RD()GTATK()ATERLA-Aad-FLQRS SF-Gly(Me)-A- (Compound 34) lle(Me)-PSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-RD()GTATK()ATERLA-Aad-FLQRS SF-Gly(Me)-A- (Compound 35) lle(Me)-LSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-RD()GTATK()ATERLA-Aad-FLQRS SF-Gly(Me)-A- (Compound 3 6) lle(Me)-LSSTETGSNT-Hyp-NH2 [ 19CD]-isoGlu-ED()GTATK()ATERLA-Aad-FLQRSSFGly(Me)-A- (Compound 3 7) lle(Me)-LSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-RD()GTATK()ATERLA-Aad-FLQRTTF-Gly(Me)-A- (Compound 3 8) lle(Me)-LSSTEVGSNT-Hyp-NH2 [19CD]-isoGlu-KD()GTATK()ATQRLA-Aad-FLQRSSF-Gly(Me)- (Compound 39) Alle(Me)-LSSTEVGSNTHyp-NH2 [19CD]-isoGlu-RD()GTATK()ATQRLA-Aad-FLQRSSF-Gly(Me)-A- (Compound 40) lle(Me)-LSSTEVGSNT-Hyp-NH2 [19CD]-isoGlu-RD()GTATK()ATQRLADFLQRSSF-Gly(Me)-A- (Compound 41) lle(Me)-LSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-RD()GTATK()ATQRLADFLQRSSF-Gly(Me)-A- (Compound 42) lle(Me)-LSSTETGSNT-Hyp-NH2 [19CD]-isoGlu-KD()GTATK()ATQRLANFLQRSSF-Gly(Me)-A- (Compound 43) lle(Me)-LSSTEVGSNT-Hyp-NH2 [ 19CD]-isoGlu-KD()GTATK()ATQRLANFLQRSSF-Gly(Me)-A- (Compound 44) lle(Me)-LSSTETGSNT-Hyp-NH2 [19CD]-isoGlu-R-Dap()-GTAT-Aad()-ATERLAHFLQRSSF-Gly(Me)- (Compound 48) A-lle(Me)-LSSTEVGSNT-Hyp-NH ₂ or - 35 042757 [19CD]-isoGlu-RE()GTAT-Dab()-ATERLAHFLQRSSF-Gly(Me)-A- (Compound 51) lle(Me)-LSSTEVGSNT-Hyp-NH ₂ or a pharmaceutically acceptable salt or solvate thereof; where parentheses () represent an intramolecular lactam bridge formed between the side chains of residues; [19CD] is a 19-carboxynodecanoyl group; isoGlu is an isoglutamic acid residue; Dab is 2,4-diaminobutanoic acid; Gly(Me) is N-methylglycine; Pe(Me) is N-methylisoleucine; Nur is 4-hydroxyproline; From is ornithine; hLys is 2-amino-7-aminoheptanoic acid (homolysin); Aad is 2-aminoadipic acid and Dap is 2,3-diaminopropanoic acid. 2. Amylin analog according to claim 1 having the formula [19CD]-isoGlu-RD()GTATK()ATERLA-Aad-FLQRSSF-Gly(Me)-A-lle(Me)-LSSTEVGSNT-Hyp-NH2 (Compound 35) or a pharmaceutically acceptable salt or solvate thereof. 3. A pharmaceutical composition comprising an amylin analog according to claim 1 or 2 in combination with a pharmaceutically acceptable carrier, excipient or carrier. 4. A method for synthesizing an amylin analog according to claim 1 or 2, comprising synthesizing said analog using a solid phase or liquid phase peptide synthesis technique, optionally including isolation and/or purification of the end product, and optionally further comprising the step of forming an amide bond between the side chains at positions 2 and 7. 5. Use of an amylin analogue according to claim 1 or 2 in a method of therapeutic treatment. 6. Use according to claim 5 for the treatment of obesity, morbid obesity, pre-surgical obesity, obesity-related inflammation, obesity-related gallbladder disease, and obesity-induced sleep apnea and respiratory disorders, cartilage degeneration, osteoarthritis, or reproductive complications. health associated with obesity or overweight. 7. Use according to claim 5 for the prevention or treatment of Alzheimer's disease, diabetes, type 1 diabetes, type 2 diabetes, prediabetes, insulin resistance syndrome, impaired glucose tolerance (IGT), pathological conditions associated with elevated blood glucose, metabolic disease, including metabolic syndrome, hyperglycemia, hypertension, atherogenic dyslipidemia, hepatic steatosis (fatty liver; including non-alcoholic fatty liver disease (NAFLD), which in turn includes non-alcoholic steatohepatitis (NASH)), renal failure, arteriosclerosis (eg, atherosclerosis ), large vessel disease, small vessel disease, diabetes-induced heart disease (including diabetic cardiomyopathy and heart failure as a complication of diabetes), coronary heart disease, peripheral arterial disease or stroke, and combinations thereof. 8. Use according to claim 5 to reduce circulating LDL and/or increase HDL/LDL ratio. 9. The use of an amylin analog according to claim 1 or 2 in a method of treating, slowing or limiting weight gain, promoting weight loss and/or reducing excess body weight. 10. Use according to any one of claims 6 to 9, wherein said amylin analog is administered as part of a combination therapy together with an antidiabetic agent, an antiobesity agent, a metabolic syndrome agent, an antidyslipidemic agent, an antihypertensive agent, a proton pump inhibitor, or an anti-inflammatory agent. 11. Use according to claim 10, wherein said anti-obesity agent is a peptide YY, a neuropeptide Y (NPY), a cannabinoid 1 receptor antagonist, a lipase inhibitor, a human proislet peptide (HIP), a melanocortin 4 receptor agonist, a GLP-1 receptor agonist, melanin concentrating hormone 1 receptor antagonist, CCK, amylin, pramlintide, or leptin. Eurasian Patent Organization, EAPO Russia, 109012, Moscow, Maly Cherkassky per., 2