TW201315477A - Polypeptides - Google Patents

Abstract

The invention relates to polypeptides comprising an amino acid sequence which is an analogue of pramlintide, pharmaceutical compositions comprising these polypeptides, and these polypeptides for use as medicaments.

Description

Polypeptide

The present invention relates to a multi-peptide comprising the amino acid sequence of SEQ ID No: 2 (pramlintide) analog; a pharmaceutical composition comprising the multi-peptide; and such a pharmaceutical product Peptide.

The number of people suffering from diabetes and obesity is growing and growing. Diabetes is a metabolic disorder in which the ability to utilize glucose is partially or completely lost.

Many treatments target hyperglycemia, while other treatments focus on weight loss. The most effective anti-diabetic agents used to lower blood sugar are insulin and its analogs. It has long been known that weight is increased when using traditional insulin to treat diabetes. Insulin must be injected subcutaneously several times a day.

Type 2 diabetes is usually treated with diet and exercise at an early stage. As the disease progresses, different oral anti-diabetic agents are added. Injectable agents, such as GLP-1 analogs, can also be used at this stage. These agents are generally most effective for the patient, allowing beta-cells that release insulin and amylase to function.

Human amyloid (SEQ ID No: 1) is a 37 amino acid long polypeptide having physicochemical properties that make it difficult to use as a drug. In particular, amyloid tends to undergo fibril formation (i.e., fibrils) in vitro and/or in vitro, and becomes ineffective due to precipitation. In addition, amyloid is difficult to formularate due to its chemical instability and precipitation at physiological pH. Therefore, amylose is formulated in an acidic solution.

Human amyloid binds to two different receptor complexes. These two complexes contain the calcitonin receptor plus receptor activity modifying protein RAMP1 or RAMP3. From the close relationship between the calcitonin receptor and the amyloid receptor, some cross-reactivity of the amyloid receptor agonist to the calcitonin receptor can be expected. For example, pramlintide has some affinity for the calcitonin receptor, but has a 14-fold affinity for the amyloid receptor.

Calcitonin receptors are found in many tissues throughout the body and are involved in the regulation of bone metabolism. Salmon calcitonin is currently marketed under the trademark Miacalcic®. This product is used for hypercalcemia, osteoporosis (including postmenopausal osteoporosis and glucocorticoid-related osteoporosis), malformation osteitis (Pagets disease), and daily injection Or nasally once. Calcitonin binds to specific receptors in the periosteum, kidney, and central nervous system (CNS). The plasma half-life of salmon calcitonin is about 45 minutes.

The multi-peptide which is active against the calcitonin receptor can be used for the treatment of hypercalcemia, osteoporosis, Paget's disease, obesity or obesity-related diseases and prevention of obesity-related diseases. The disadvantage of treatment with the currently used calcitonin preparation is that since the plasma half-life of salmon calcitonin is short, the drug must be administered several times a day and must be administered immediately after the meal.

A multi-peptide that has dual activity on both the calcitonin receptor and the amyloid receptor can be advantageous.

Pramlintide (SEQ ID No: 2) as to Amylin Pharmaceuticals Symlin ^® marketed drugs for the treatment of diabetes drug as an adjunct to insulin. Pramlintide is an amyloid receptor agonist. Its activity on calcitonin receptors is as small as about one-fourth.

The chemical structure of pramlintide is presented below and in Figure 1.

The pramlintide is chemically unstable at neutral pH and it is therefore provided in an acidic solution. The amino acids at positions 25, 28 and 29 in pramlintide are substituted with proline by comparison with human amyloid. This modification reduces the tendency of the protein to undergo fibril formation. Pramlintide has a very short plasma half-life and therefore must be injected two to three times a day.

WO2010046357 and WO2009034119 disclose polypeptides (herein referred to as amyloid derivatives) comprising an amyloid analog having an albumin binding residue. Although these multi-peptides with albumin binding moieties exhibit improved pharmacokinetic (PK) or pharmacodynamic (PD) properties compared to pramlintide, they may still exhibit poorer physics under certain conditions. stability.

It has been surprisingly found that a multi-peptide comprising an amino acid of SEQ ID No: 2 (pramlintide) analogue may demonstrate an increase in solubility and/or physical stability, the amino acid comprising glutamic acid at position 14. Amino acid residue.

In at least some embodiments, the multi-peptide of the invention has increased solubility.

In at least some embodiments, the multi-peptide of the invention has increased physical stability.

In at least some embodiments, the multi-peptide of the invention has increased solubility and physical stability.

In at least some embodiments, the multi-peptides of the invention exhibit advantageous pharmacokinetic characteristics and/or advantageous pharmacological characteristics. An example of a favorable pharmacokinetic profile is a long-lasting feature.

In a broad aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein the analog comprises a glutamic acid residue at position 14, wherein the analog The amino acid sequence number corresponds to the amino acid numbering sequence of SEQ ID No: 2, and the restriction is that the multi-peptide is not N-α-(19-carboxynonanoyl)-Glu-Glu-Arg- [Glu1, Glu14, His17, Glu31]-planin. Human amylin receptor potency assay (such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than ₅₀ of the EC 1800 pM. Human calcitonin receptor potency assay (such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than 1800 pM of EC _50. In solubility assays (such as disclosed herein), the multi-peptide has a solubility of about 100 μM or greater than 100 μM, as appropriate at pH 4. In solubility assays (such as disclosed herein), the multi-peptide has a solubility of about 100 μM or greater than 100 μM, as appropriate at pH 7. In a fibril formation assay (also known as a fibrillation assay) such as disclosed herein, the multi-peptide has a physical stability of about 25 hours or more than 25 hours, as appropriate. The multipeptide is optionally attached to at least one of its at least one amino acid residue. The invention also relates to pharmaceutical formulations comprising the same. The invention also relates to its medical use. The invention also relates to the delivery (such as administration) to a patient in need of treatment thereof.

In another broad aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein the analog comprises a glutamic acid residue at position 14 and a position 17 A histidine residue, wherein the amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2, and the restriction is that the polypeptide is not N-α-(19-carboxyl Nine fluorenyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]-pramlintide. Human amylin receptor potency assay (such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than ₅₀ of the EC 1800 pM. Human calcitonin receptor potency assay (such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than 1800 pM of EC _50. In solubility assays (such as disclosed herein), the multi-peptide has a solubility of about 100 μM or greater than 100 μM, as appropriate at pH 4. In solubility assays (such as disclosed herein), the multi-peptide has a solubility of about 100 μM or greater than 100 μM, as appropriate at pH 7. In a fibril formation assay (also known as a fibrillation assay) such as disclosed herein, the multi-peptide has a physical stability of about 25 hours or more than 25 hours, as appropriate. The multipeptide is optionally attached to at least one of its at least one amino acid residue. The invention also relates to pharmaceutical formulations comprising the same. The invention also relates to its medical use. The invention also relates to the delivery (such as administration) to a patient in need of treatment thereof.

In another broad aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein the analog comprises a glutamic acid residue at position 14 and a position 17 A arginine residue wherein the amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2. Human amylin receptor potency assay (such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than ₅₀ of the EC 1800 pM. Human calcitonin receptor potency assay (such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than 1800 pM of EC _50. In solubility assays (such as disclosed herein), the multi-peptide has a solubility of about 100 μM or greater than 100 μM, as appropriate at pH 4. In solubility assays (such as disclosed herein), the multi-peptide has a solubility of about 100 μM or greater than 100 μM, as appropriate at pH 7. In a fibril formation assay (also known as a fibrillation assay) such as disclosed herein, the multi-peptide has a physical stability of about 25 hours or more than 25 hours, as appropriate. The multipeptide is optionally attached to at least one of its at least one amino acid residue. The invention also relates to pharmaceutical formulations comprising the same. The invention also relates to its medical use. The invention also relates to the delivery (such as administration) to a patient in need of treatment thereof.

In another broad aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein the analog comprises a glutamic acid residue at position 14 and a position 37 A proline residue in which the amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2. Human amylin receptor potency assay (such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than ₅₀ of the EC 1800 pM. Human calcitonin receptor potency assay (such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than 1800 pM of EC _50. In solubility assays (such as disclosed herein), the multi-peptide has a solubility of about 100 μM or greater than 100 μM, as appropriate at pH 4. In solubility assays (such as disclosed herein), the multi-peptide has a solubility of about 100 μM or greater than 100 μM, as appropriate at pH 7. In a fibril formation assay (also known as a fibrillation assay) such as disclosed herein, the multi-peptide has a physical stability of about 25 hours or more than 25 hours, as appropriate. The multipeptide is optionally attached to at least one of its at least one amino acid residue. The invention also relates to pharmaceutical formulations comprising the same. The invention also relates to its medical use. The invention also relates to the delivery (such as administration) to a patient in need of treatment thereof.

In another broad aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein the analog comprises a glutamic acid residue at position 14 and a position 35 A histidine residue wherein the amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2. Human amylin receptor potency assay (such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than ₅₀ of the EC 1800 pM. Human calcitonin receptor potency assay (such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than 1800 pM of EC _50. In solubility assays (such as disclosed herein), the multi-peptide has a solubility of about 100 μM or greater than 100 μM, as appropriate at pH 4. In solubility assays (such as disclosed herein), the multi-peptide has a solubility of about 100 μM or greater than 100 μM, as appropriate at pH 7. In a fibril formation assay (also known as a fibrillation assay) such as disclosed herein, the multi-peptide has a physical stability of about 25 hours or more than 25 hours, as appropriate. The multipeptide is optionally attached to at least one of its at least one amino acid residue. The invention also relates to pharmaceutical formulations comprising the same. The invention also relates to its medical use. The invention also relates to the delivery (such as administration) to a patient in need of treatment thereof.

In another broad aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein the analog comprises a glutamic acid residue at position 14 and a position 17 A histidine or arginine residue and a proline residue at position 37, wherein the analog amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2. Human amylin receptor potency assay (such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than ₅₀ of the EC 1800 pM. Human calcitonin receptor potency assay (such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than 1800 pM of EC _50. In solubility assays (such as disclosed herein), the multi-peptide has a solubility of about 100 μM or greater than 100 μM, as appropriate at pH 4. In solubility assays (such as disclosed herein), the multi-peptide has a solubility of about 100 μM or greater than 100 μM, as appropriate at pH 7. In a fibril formation assay (also known as a fibrillation assay) such as disclosed herein, the multi-peptide has a physical stability of about 25 hours or more than 25 hours, as appropriate. The multipeptide is optionally attached to at least one of its at least one amino acid residue. The invention also relates to pharmaceutical formulations comprising the same. The invention also relates to its medical use. The invention also relates to the delivery (such as administration) to a patient in need of treatment thereof.

In another broad aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein the analog comprises a glutamic acid residue at position 14 and a position 17 A histidine or arginine residue and a histidine residue at position 35, wherein the analog amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2. Human amylin receptor potency assay (such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than ₅₀ of the EC 1800 pM. Human calcitonin receptor potency assay (such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than 1800 pM of EC _50. In solubility assays (such as disclosed herein), the multi-peptide has a solubility of about 100 μM or greater than 100 μM, as appropriate at pH 4. In solubility assays (such as disclosed herein), the multi-peptide has a solubility of about 100 μM or greater than 100 μM, as appropriate at pH 7. In a fibril formation assay (also known as a fibrillation assay) such as disclosed herein, the multi-peptide has a physical stability of about 25 hours or more than 25 hours, as appropriate. The multipeptide is optionally attached to at least one of its at least one amino acid residue. The invention also relates to pharmaceutical formulations comprising the same. The invention also relates to its medical use. The invention also relates to the delivery (such as administration) to a patient in need of treatment thereof.

In another broad aspect, the invention relates to a multi-peptide comprising an amino acid sequence which is an analog of SEQ ID No: 2 of formula (I): Xaa ₁ -Cys-Xaa ₃ -Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Glu-Phe-Leu-Xaa ₁₇ -Xaa ₁₈ -Ser-Ser-Xaa ₂₁ -Xaa ₂₂ -Phe-Gly-Pro-Xaa ₂₆ -Leu-Pro-Pro-Thr-Xaa ₃₁ -Val-Gly-Ser-Xaa ₃₅ -Thr-Xaa ₃₇ ; Formula (I) (SEQ ID No: 3)

Wherein Xaa _{1 is} deleted or independently selected from the group consisting of Ala, Cys, Glu, Gly, His, Arg, Ser and Lys; Xaa _{3 is} independently selected from Gly, His, Arg, Ser and Asn; Xaa _{17 is} independently selected from His, Arg , Lys and Val; Xaa _{18 is} independently selected from Arg, Lys and His; Xaa _{21 is} independently selected from the group consisting of Ala, Lys, Gln, Ser and Asn; Xaa _{22 is} independently selected from Glu, Gln, Ser, Thr and Asn; Xaa _{26 is} independently selected from the group consisting of Pro, Arg and Ile; Xaa _{31 is} independently selected from the group consisting of Ser, Glu, Asp and Asn; Xaa _{35 is} independently selected from the group consisting of His, Arg, Lys, Asp and Glu; Xaa _{37 is} independently selected from Pro and Tyr And wherein the C-terminus can be derivatized as appropriate; and the restriction condition is that the multi-peptide is not N-α-(19-carboxynonanoyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31] - pramlintide; human amylin receptor potency assay (such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than 1800 pM of EC _50; calcitonin receptor assay potency in humans ( such as disclosed herein), the multi-peptide optionally having about 1800 pM, or less than 1800 pM of EC _50.

In a solubility assay (such as disclosed herein), the multi-peptide has a solubility of about 100 μM or greater than 100 μM, optionally at pH 4; in a solubility assay (such as disclosed herein), the multi-peptide is The pH 7 has a solubility of about 100 μM or greater than 100 μM as appropriate; in a fibril formation assay (also known as a fibrillation assay) such as disclosed herein, the multi-peptide has about 25 hours or more depending on the situation. 25 hours of physical stability; the multipeptide is optionally attached to at least one of its at least one amino acid residue.

The invention also relates to pharmaceutical formulations comprising the same. The invention also relates to Medical use. The invention also relates to the delivery (such as administration) to a patient in need of treatment thereof.

In another aspect, the invention further comprises a pharmaceutical composition comprising the above multi-peptide.

In another aspect, the invention further comprises a method for preparing a pharmaceutical composition comprising the above multi-peptide.

In another aspect, the invention further comprises the above-described multi-peptides useful as pharmaceuticals.

The multipeptides of the invention are advantageous for their improved solubility and/or physical stability.

Described herein are assays for determining the potency of the amyloid receptor and calcitonin receptor and determining the solubility and physical stability of the multipeptide. For example, see tests (II), (IV) and (III), respectively.

definition

The term " human amylin " as used herein relates to a multi-peptide human amyloid having the sequence set forth in SEQ ID No. 1. The term includes, but is not limited to, a human polypeptide hormone having 37 amino acids known as amyloid, which is substantially co-secreted by pancreatic β-cells together with insulin. Human amyloid has the following primary amino acid sequence: Lys-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Asn-Phe-Leu-Val-His-Ser-Ser -Asn-Asn-Phe-Gly-Ala-Ile-Leu-Ser-Ser-Thr-Asn-Val-Gly-Ser-Asn-Thr-Tyr ( SEQ ID NO: 1 )

Human amyloid has a disulfide bridge between the two Cys residues and a C-terminal guanamine group. This structure is shown below and in Figure 4.

Herein, SEQ ID No: 1 and human amyloid are used interchangeably.

The term " pramlintide " as used herein relates to a synthetic multi-peptide having the sequence set forth in SEQ ID No. 2. Pramlintide has the following primary amino acid sequence: Lys-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Asn-Phe-Leu-Val-His-Ser-Ser -Asn-Asn-Phe-Gly-Pro-Ile-Leu-Pro-Pro-Thr-Asn-Val-Gly-Ser-Asn-Thr-Tyr (SEQ ID NO: 2)

Pramlintide has a disulfide bridge between the two Cys residues and a C-terminal guanamine group. This structure is shown below and in Figure 1.

Herein, SEQ ID No: 2 and pramlintide are used interchangeably.

The term " calcitonin " means salmon calcitonin or human calcitonin.

The term " salmon calcitonin " or " sCT " means the natural protein sequence of salmon calcitonin as disclosed in Niall et al. (1969), Biochemistry, Vol. 64 , Figure 2. Salmon calcitonin is a multi-peptide consisting of 32 amino acids. It has a disulfide bridge between the first amino acid and the seventh amino acid at the amino terminal of the poly-peptide, which is necessary for its biological activity and has a carboxyl terminal amino acid Amidoxime.

The term " human calcitonin " means the native protein sequence of human calcitonin as disclosed in Niall et al. (1969), Biochemistry, Vol. 64 , Figure 2. Human calcitonin is a multi-peptide consisting of 32 amino acids. It has a disulfide bridge between the first amino acid and the seventh amino acid at the amino terminal of the poly-peptide, which is necessary for its biological activity and has a carboxyl terminal amino acid Amidoxime.

The term " analogue of amylin or amylin analogue " as used herein refers to a variant of SEQ ID No: 1.

The term " analogue of pramlintide or pramlintide analogue " as used herein refers to a variant of SEQ ID No: 2.

For example, such variants include, but are not limited to, one or more substitutions and/or ones of any one of the amino acid residues of any natural or unnatural amino acid, synthetic amino acid or peptidomimetic Any of a plurality of deletions and/or one or more additions, and/or any of natural or unnatural amino acids, synthetic amino acids or peptidomimetics, may be attached at any available position.

The variant may have the same number of amino acid residues as pramlintide (i.e., 37). Alternatively, the variant may comprise fewer amino acid residues than pramlintide. Alternatively, the variant may comprise more amino acid residues than pramlintide. In some embodiments, the variant may have the same number of amino acid residues as pramlintide (i.e., 37). In some embodiments, the variant includes any substitution of an amino acid residue of any natural or unnatural amino acid, synthetic amino acid or peptidomimetic, and/or a natural or unnatural amino acid, a synthetic amine Any of the base acid or peptidomimetic has a substituent attached to any available position.

The multi-peptide may comprise one or more amino acid substitutions. So for some In a specific example, the number of amino acid substitutions in the amyloid analog may be at least one. The number of amino group substitutions is preferably from 1 to 15, more preferably from 1 to 12, still more preferably from 1 to 10, still more preferably from 1 to 5, still more preferably from 1 to 3.

The number of amino acid insertions, additions, deletions or substitutions may be at least 1, but up to 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid insertions, additions, deletions or substitutions may be present . Any natural or unnatural amino acid, synthetic amino acid, peptidomimetic or other compound may be substituted or added. The addition or deletion of an amino acid residue can occur at the N-terminus of the peptide and/or at the C-terminus of the peptide.

As used herein, the term " natural amino acid " is an amino acid selected from the group consisting of: commonly used three letter codes & one-letter codes in brackets: glycine (Gly & G), proline (Pro & P), alanine (Ala & A), valine (Val & V), leucine (Leu & L), isoleucine (Ile & I), methyl sulfide Amine acid (Met & M), cysteine (Cys & C), phenylalanine (Phe & F), tyrosine (Tyr & Y), tryptophan (Trp & W), histidine (His & H), lysine (Lys & K), arginine (Arg & R), glutamic acid (Gln & Q), aspartic acid (Asn & N), glutamic acid (Glu & E) Aspartic acid (Asp & D), serine (Ser & S) and threonine (Thr & T). If the font does not match the usual code due to a typing error, the common code is applied. The amino acid present in the multipeptide of the present invention is preferably an amino acid which can be encoded by a nucleic acid.

If the analog contains more than 37 amino acid residues or less than 37 amino acid residues, those skilled in the art can still determine the corresponding sequence with the pramlintide sequence (SEQ ID No. 2). The position number of the amino acid residue. The appropriate comparison program is "needle", which is Needleman-Wunsch ratio Correct. The algorithm for this alignment program is described in Needleman, S. B. and Wunsch, CD., (1970), Journal of Molecular Biology, 48: 443-453.

In the numbered sequence of SEQ ID No: 2, and according to the practice in the art, the N-terminal amino acid residue (Lys) is designated as No. 1 and the subsequent amino acid residues are numbered sequentially, terminating at the C-terminus. Designated as No. 37 tyrosine. Thus, any reference herein to the amino acid residue position numbering generally provides for its position in the 37 amino acid sequences; the 37 amino acid sequences are analogs of pramlintide. For example, reference to an analog modified at position 14 can refer to an analog wherein the 14th amino residue of the 37 amino acids of the analog has been modified.

In other words, the amino acid sequence numbering of the analogs provides the position of each analog with respect to the 37 amino acid sequences, wherein the numbering is in the ascending order of the N-terminus to the C-terminus.

Analogs can be described by reference to the numbering (i.e., its position) of the modified amino acid residue in pramlintide or human amyloid and the nature of the modification. The following are non-limiting examples of the appropriate analog nomenclature.

For example: [Glu14]-pramlintide represents an analog of SEQ ID No: 2 (pramlintide) wherein the change in pramlintide is such that Asn at position 14 is substituted with Glu.

[Glu14, Arg17, His35]-pramlintide represents an analog of SEQ ID No: 2 (pramlintide) in which Asn at position 14 has been replaced by Glu, Val at position 17 has been substituted by Arg, and position 35 Asn has been replaced by His.

In another example, des1 (or Des ¹ ) with respect to the pramlintide analog refers to an analog in which the N-terminal amino acid is dehydrated from the amino acid. A pramlintide analog in which the N-terminal amino acid has been deleted may also be referred to as des1 pramlintide.

[Pro25, Pro28, Pro29] - Human amyloid represents an analog of SEQ ID No: 1 (human amyloid) in which human amyloid is modified such that Ala at position 25 and Cys at positions 28 and 29 have been replaced by Pro . This multipeptide is pramlintide. Therefore, it is understood that pramlintide is a human amyloid analog. Therefore, "pramlintide analog" is used interchangeably with "amyloid analog".

As is apparent from the above examples, the amino acid residue can be identified by its full name, its one-letter code, and/or its three-letter code. These three methods are completely equivalent.

As used herein, the expression "conforms to", "corresponds to", "a position equivalent to" or "corresponding position" may be used by reference to SEQ. ID No: 2 to characterize the modification site in the pramlintide analog. Equivalent or corresponding positions are readily deduced, for example, by simple writing and visual inspection; and/or standard protein or multi-peptide alignment programs, such as "needle", which is a Needleman-Wunsch alignment, can be used. The algorithm is described in Needleman, SB and Wunsch, CD., (1970), Journal of Molecular Biology, 48: 443-453, and the alignment program is described by Myers and W. Miller in "Optimal Alignments in Linear Space" CABIOS (computer Applications in the biosciences) (1988) 4:11-17. For alignment, a pre-designed sub-matrix BLOSUM62 and a preset unit matrix can be used, and the gap penalty for the first residue can be set to -10 and the gap penalty for other residues can be set to -0.5.

The multi-peptide may comprise one or more substituents on one or more amino acid residues. These multi-peptides may also be referred to as pramlintide derivatives or amyloid derivatives.

The term " substituent " as used herein means any suitable moiety of a bond (particularly covalently bonded) to an amino acid residue, particularly an amino acid residue, at any available position. Suitable portions are typically chemical moieties.

For some specific examples, the substituents comprise a linker.

For some embodiments, the multipeptide has a substituent on an amino acid residue that is an amino acid residue at the N-terminal residue or the amino acid residue is an lysine.

For some specific examples, the multi-peptide has a substituent bonded via an a-amino group of an N-terminal amino acid residue on an N-terminal amino acid residue.

For some specific examples, the N-terminal amino acid residue is an amine acid and the polypeptide has a substituent bonded to the ε-amine group via an amino acid amine residue on the N-terminal amino acid residue.

For some specific examples, the multi-peptide is extended by the addition of an lysine residue at the N-terminus, and the multi-peptide has an epsilon-amino group via an amino acid residue of the amino acid group at the N-terminal amino acid residue. A combination of substituents.

For some specific examples, the multi-peptide is extended by the addition of an amino acid residue at the N-terminus, and the multi-peptide has an α-amino group via an N-terminal amino acid residue on the N-terminal amino acid residue. A combination of substituents.

The term " hydrocarbyl " as used herein refers to a group containing at least carbon and hydrogen and optionally one or more other suitable substituents. Examples of such substituents may include a hydroxyl group, an alkyl group, a halogen group, an alkoxy group, a haloalkyl group, a haloalkoxy group, an amine group, an aminoalkyl group or a cyclic group. In addition to the possibility that the substituent is a cyclic group, a combination of substituents can form a cyclic group. If the hydrocarbyl group contains more than one carbon atom, then the carbon atoms are not necessarily linked to each other. For example, at least two carbon atoms can be attached via a suitable atom or group. Therefore, the hydrocarbon group may contain a hetero atom. Suitable heteroatoms will be apparent to those skilled in the art and include, for example, sulfur, nitrogen, oxygen, phosphorus, and antimony. In one embodiment, the hydrocarbyl group is selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, heteroaryl or cycloalkyl, each of which may be substituted as appropriate. Examples of such substituents may include hydroxy, alkyl, halo, alkoxy, haloalkyl, haloalkoxy, amine, aminoalkyl or cycloalkyl.

The term " alkyl " as used herein includes saturated straight-chain and branched-chain alkyl groups which may be substituted (mono- or poly-substituted) or unsubstituted. The alkyl group is preferably a C _1-20 alkyl group, more preferably a C _1-15 alkyl group, more preferably a C _1-10 alkyl group, more preferably a C _1-8 alkyl group, more preferably a C _1-6 alkyl group. Particularly preferred alkyl groups include, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl and Zheng Xinji. Suitable substituents include, for example, hydroxy, alkyl, halo, alkoxy, haloalkyl, haloalkoxy, amine, aminoalkyl or cycloalkyl.

The term " cycloalkyl " as used herein refers to a cyclic alkyl group which may be substituted (monosubstituted or polysubstituted) or unsubstituted. Suitable substituents include, for example, hydroxy, alkyl, halo, alkoxy, haloalkyl, haloalkoxy, amine, aminoalkyl or cycloalkyl.

The term " alkenyl " as used herein, refers to a carbon chain containing one or more carbon-carbon double bonds which may be branched or unbranched and substituted (monosubstituted or polysubstituted) or unsubstituted. . The alkenyl group is preferably a C _2-20 alkenyl group, more preferably a C _2-15 alkenyl group, more preferably a C _2-10 alkenyl group, more preferably a C _2-8 alkenyl group or a more preferably a C _2-6 alkenyl group. Suitable substituents include, for example, hydroxy, alkyl, halo, alkoxy, haloalkyl, haloalkoxy, amine, aminoalkyl or cycloalkyl.

The term " alkynyl " as used herein, refers to a carbon chain containing one or more carbon-carbon bonds, which may be branched or unbranched, substituted (monosubstituted or polysubstituted) or unsubstituted. . The alkynyl group is preferably a C _2-20 alkynyl group, more preferably a C _2-15 alkynyl group, more preferably a C _2-10 alkynyl group, more preferably a C _2-8 alkynyl group or a more preferably a C _2-6 alkynyl group. Suitable substituents include, for example, hydroxy, alkyl, halo, alkoxy, haloalkyl, haloalkoxy, amine, aminoalkyl or cycloalkyl.

The term " aryl " as used herein refers to a _C6-10 aromatic group which may be substituted (monosubstituted or polysubstituted) or unsubstituted. Typical examples include phenyl, naphthyl and the like. Suitable substituents include, for example, hydroxy, alkyl, halo, alkoxy, haloalkyl, haloalkoxy, amine, aminoalkyl or cycloalkyl.

The term " heteroaryl " as used herein refers to an aryl group as defined above containing one or more heteroatoms. Suitable heteroatoms will be apparent to those skilled in the art and include, for example, sulfur, nitrogen, oxygen, phosphorus, and antimony. Suitable substituents include, for example, hydroxy, alkyl, halo, alkoxy, haloalkyl, haloalkoxy, amine, aminoalkyl or cycloalkyl.

The term " linker " as used herein includes suitable substituents that allow a moiety, such as a chemical moiety, to be joined to a multi-peptide (such as a multi-peptide backbone). Therefore, the linker and the chemical moiety together become a substituent. The portion joined to the linker can be any suitable portion. Examples include albumin binding moieties.

In one embodiment, the albumin binding moiety has from 6 to 40 carbon atoms, from 8 to 26 carbon atoms, or from 14 to 22 carbon atoms, such as 16, 17, 18, 19, 20 carbon atoms.

In another embodiment, the albumin binding moiety is a thiol group selected from the group consisting of CH ₃ (CH ₂ ) _r CO-, wherein r is an integer from 4 to 38, preferably an integer from 4 to 24; more preferably Selected from the group consisting of CH ₃ (CH ₂ ) ₆ CO-, CH ₃ (CH ₂ ) ₈ CO-, CH ₃ (CH ₂ ) ₁₀ CO-, CH ₃ (CH ₂ ) ₁₂ CO-, CH ₃ (CH ₂ ) ₁₄ A group of CO-, CH ₃ (CH ₂ ) ₁₆ CO-, CH ₃ (CH ₂ ) ₁₈ CO-, CH ₃ (CH ₂ ) ₂₀ CO-, and CH ₃ (CH ₂ ) ₂₂ CO-.

In a specific example, the albumin binding moiety comprises a group that can be negatively charged at pH 7.4.

In one particular example, the albumin binding moiety containing a carboxylic acid group, such as HOOC (CH _{2) s} CO-, wherein s is an integer of 12 to 22. s is preferably 16 or 18.

In a specific example, the portion joined to the linker is an albumin binding moiety.

For example, a linker can comprise one or two amino acids that bind to a moiety (such as an albumin binding moiety) at one end and to any available position on the polypeptide backbone at the other end.

In some embodiments, the linker provides a bridge or linkage between the amine group on the polypeptide backbone and a thiol group on a moiety, such as an albumin binding moiety. The linker can be attached to or near the N-terminal amino acid residue. The linker preferably binds to the amino acid of position 1 of the pramlintide analog.

Another example of a linker is a combination of at least one amino acid and an amine.

In one embodiment, the amine is preferably a group OEG, wherein the chemical formula of OEG is shown below:

For some specific examples, the linker is preferably selected from the group consisting of γGlu, γGlu-γGlu, γGlu-γGlu-γGlu, γGlu-γGlu-γGlu-γGlu, Glu, Glu-Glu, Glu-γGlu, Glu-Arg, Glu-Glu-Arg, His, His-His, His-γGlu, His-His-γGlu, Gly, Gly-γGlu, Ser, Ser-γGlu, D-Arg-D-Arg, Arg, Arg-Arg, Arg- Arg-γGlu, Ser-Ser, -Gly-Ser-Ser, Ser-Ser, -Gly-Ser-Ser-γGlu, Ser-Ser-Gly-Ser-Ser-Gly and Ser-Ser-Gly-Ser-Ser- Gly-γGlu, γGlu-OEG, γGlu-2xOEG and OEG, the linker is preferably selected from the group consisting of γGlu, γGlu-γGlu, γGlu-OEG, γGlu-2xOEG and OEG, and the linker is more preferably γGlu-γGlu.

The linker may contribute to and/or enhance the binding effect of a portion (eg, an albumin binding moiety), for example, a linker comprising γGlu may enhance the albumin binding effect of the multipeptide.

By the use of the terms "γ Glu " or " gGlu " or γ Glu or γ -L-Glu means an amino acid having the following structure (also shown in Figure 2):

Α-nitrogen and γ-carboxyl form a guanamine bond that binds to two adjacent residues

The use of the term "γ Glu- γ Glu " means a part having the following structure:

The use of the term "γ Glu-OEG " means the part with the following structure:

The use of the term "γ Glu-OEG-OEG " means the part with the following structure:

The term " epilson amino group or ε- amino group " as used herein with respect to lysine refers to an amine group specified at the 6 position using the IUPAC standard numbering. The term " alpha amino group or alpha- amino group " refers to an amine group specified at the 2-position using the IUPAC standard numbering. See the structure below (also shown in Figure 3).

The term " albumin binding moiety " as used herein refers to a chemical group capable of binding to albumin (ie, having albumin binding affinity). In a specific example, the albumin binding moiety is a sulfhydryl group.

In some embodiments, the albumin binding moiety is preferably a thiol group selected from the group consisting of: (a) CH ₃ (CH ₂ ) _r CO-, wherein r is an integer from 4 to 24; (b) HOOC (CH ₂ ) _s CO-, where s is an integer from 14 to 20, such as 16 or 18.

" Albumin binding affinity " can be determined by several methods known in the art. In one method, the compound to be measured is radiolabeled with, for example, ¹²⁵ I or ³ H and incubated with immobilized albumin (Kurtzhals et al, Biochem. J., 312, 725-731 (1995)). Calculate the binding of the compound to the standard. In another method, the related compound is radiolabeled and it competes with a series of diluted compounds to be measured for binding to albumin immobilized, for example, on SPA beads. EC ₅₀ value is a measure of competition compound affinities. In a third method, the receptor affinity or potency of a compound is measured under different concentrations of albumin, and the relative affinity or potency of the compound associated with albumin concentration reflects its affinity for albumin.

The multi-peptide of the present invention shows good performance. The term " potency " is used to describe the effect of a given compound in an assay in which the sigmoidal relationship between logarithmic concentration and compound effect has been determined. In addition, the reaction can vary from 0 to 100%. EC (effective concentration) ₅₀ can be used to describe the concentration of a given compound in a assay, such as a 50% reaction in a functional assay.

The multi-peptide of the present invention shows good activity. The term " activity " refers to the ability to reduce appetite and/or increase satiety. Activity can be measured, for example, by the ability to reduce appetite as described in assay (I) herein.

The multipeptide of the present invention shows good physical stability. The term " physical stability " of a multi-peptide or a formulation thereof of the present invention means that the multi-peptide is not exposed to thermo-mechanical stress and/or to destabilizing interfaces and surfaces (such as hydrophobic surfaces and interfaces). Interactions form a tendency to biologically inactivate and/or insoluble aggregates. The physical stability of the aqueous multi-peptide formulation can be assessed by visual inspection, ThT fibrillation assay (sometimes referred to as ThT fibrillation assay), and/or turbidity measurement as described elsewhere herein. Visual inspection of the formulation was performed in sharply focused light on a dark background. The turbidity of the formulation is characterized by a visual score of turbidity, for example, on a scale of 0 to 3 (a formulation that does not exhibit turbidity corresponds to a visual score of 0, and a formulation that exhibits visible turbidity in daylight corresponds to a visual score of 3). When the formulation exhibits visible turbidity in daylight, it falls under the physical instability of protein aggregation. Alternatively, the turbidity of the formulation can be assessed by simple turbidity measurements well known to those skilled in the art.

The multipeptide of the present invention shows good chemical stability. The term " chemical stability " of a multi-peptide or a formulation thereof of the present invention means that there is no chemical covalent change in the structure of the multi-peptide, thereby avoiding the formation of a potency compared to the parent (natural) multi-peptide structure. Chemical degradation products that may reduce and/or increase immunogenicity. The type and nature of the visible parent peptides are different from the environment to which the peptides are exposed, resulting in different chemical degradation products. Excluding chemical degradation is unlikely to be completely avoided, and an increase in the amount of chemical degradation products is often found during storage and use of the multi-peptide formulations well known to those skilled in the art. Most of the multi-peptides are susceptible to deamination of amines, which are processes in which the side chain guanamine groups are hydrolyzed to form free carboxylic acids in the guanylamine sulfhydryl residues or aspartic acid guanidino residues. Other degradation pathways involve the formation of high molecular weight conversion products in which two or more multi-peptide molecules are covalently bound to each other via a transamination and/or disulfide interaction, resulting in the formation of covalently bound dimers, Polymers and polymer degradation products ( Bound of Protein Pharmaceuticals, Ahern. TJ and Manning MC, Plenum Press, New York 1992 ). Oxidation (e.g., oxidation of methionine residues) can be mentioned as another variation of chemical degradation. The chemical stability of the multi-peptide formulation can be assessed by measuring the amount of chemical degradation products at different time points after exposure to different environmental conditions (degradation product formation can often be accelerated by, for example, increasing the temperature). The amount of each individual degradation product is often determined by separating the degradation products by molecular size and/or charge using different chromatographic techniques (e.g., SEC-HPLC and/or RP-HPLC).

The term " stabilized formulation " refers to a formulation having enhanced physical stability, enhanced chemical stability, or enhanced physical and chemical stability compared to a multi-peptide aqueous solution.

General aspect

In one aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: the analog comprises a glutamic acid residue at position 14; wherein the analog amine The base acid sequence number corresponds to the amino acid number sequence of SEQ ID No: 2; The restriction condition is that the multi-peptide is not N-α-(19-carboxy nineteen -Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; The multi-peptide is optionally linked to at least one of its amino acid residues A substituent.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2; and the restriction is that the multi-peptide is not N-α-(19-carboxyundecyl)-Glu-Glu -Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; and (b) the analog has a solubility of about 100 μM or greater at pH 4, the multi-peptide is optionally at least one amine group thereof At least one substituent is attached to the acid residue.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2; and the restriction is that the multi-peptide is not N-α-(19-carboxyundecyl)-Glu-Glu -Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; and (b) the analog has a solubility of about 100 μM or greater at pH 7 The multipeptide is optionally attached to at least one of its at least one amino acid residue.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2; and the restriction is that the multi-peptide is not N-α-(19-carboxyundecyl)-Glu-Glu -Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; and (b) the multipeptide has a solubility of about 100 μM or greater at pH 4; and (c) the polypeptide is at pH 7 has a solubility of about 100 μM or greater than 100 μM. The multipeptide is optionally attached to at least one of its at least one amino acid residue.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2; and the restriction is that the multi-peptide is not N-α-(19-carboxyundecyl)-Glu-Glu -Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; and (b) the multipeptide has a solubility of about 100 μM or greater at pH 4; (c) the multi-peptide has a solubility of about 100 μM or greater than 100 μM at pH 7; and (d) the multi-peptide has a physical stability of about 25 hours or more than 25 hours in the fibril formation assay; The peptide optionally has at least one substituent attached to at least one of its amino acid residues.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The analog comprises a histidine residue at position 17; wherein the amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2; and the restriction is that the multi-peptide is not N-α-(19-carboxynonanoyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; and the multi-peptide is optionally in at least one amino acid residue thereof There is at least one substituent attached thereto.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The analog comprises a histidine residue at position 17; wherein the amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2; and the restriction is that the multi-peptide is not N-α-(19-carboxynonanoyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; (c) The analog has a solubility of about 100 μM or greater than 100 μM at pH 4. The multipeptide is optionally attached to at least one of its at least one amino acid residue.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The analog comprises a histidine residue at position 17; wherein the amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2; and the restriction is that the multi-peptide is not N-α-(19-carboxyundecyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; and (c) the multi-peptide has about 100 μM at pH 7. Or a solubility greater than 100 μM. The multipeptide is optionally attached to at least one of its at least one amino acid residue.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The analog comprises a histidine residue at position 17; wherein the amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2; and the restriction is that the multi-peptide is not N-α-(19-carboxynonanoyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; (c) the multi-peptide has a solubility of about 100 μM or more at pH 4; and (d) the multi-peptide has a solubility of about 100 μM or more at pH 7 of the multi-peptide as appropriate At least one substituent is attached to at least one of its amino acid residues.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The analog comprises a histidine residue at position 17; wherein the amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2; and the restriction is that the multi-peptide is not N-α-(19-carboxyundecyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; and (c) the multi-peptide has about 100 μM at pH 4. Or a solubility greater than 100 μM; and (d) the multipeptide has a solubility of about 100 μM or greater than 100 μM at pH 7; and (e) the multipeptide has about 25 hours or more than 25 in the fibril formation assay Physical stability of the hour; the multipeptide is optionally attached to at least one of its at least one amino acid residue.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; (b) the analog comprises a arginine residue at position 17; wherein the analog amino acid sequence number of the analog corresponds to SEQ ID No: An amino acid numbering sequence of 2; and the multi-peptide is optionally attached to at least one of its at least one amino acid residue.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The analog comprises a proline residue at position 37; wherein the amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2; and the multi-peptide is optionally at least one of At least one substituent is attached to the amino acid residue.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The analog comprises a histidine acid residue at position 35; wherein the amino acid sequence numbering of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2; and the multi-peptide is optionally at least one of At least one substituent is attached to the amino acid residue.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; (b) the analog comprises a histidine or arginine residue at position 17; (c) the analog comprises a hydrazine at position 37 An amino acid residue; wherein the amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2; and the multi-peptide is optionally linked to at least one of its at least one amino acid residue Substituent.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The analog comprises a histidine or arginine residue at position 17; (c) the analog comprises a histidine residue at position 35; wherein the analog amino acid sequence number of the analog corresponds to the SEQ ID No: 2 amino acid numbering sequence; and the multi-peptide is optionally attached to at least one of its at least one amino acid residue.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog of formula (I): Xaa ₁ -Cys-Xaa ₃ -Thr-Ala-Thr-Cys -Ala-Thr-Gln-Arg-Leu-Ala-Glu-Phe-Leu-Xaa ₁₇ -Xaa ₁₈ -Ser-Ser-Xaa ₂₁ -Xaa ₂₂ -Phe-Gly-Pro-Xaa ₂₆ -Leu-Pro-Pro- Thr-Xaa ₃₁ -Val-Gly-Ser-Xaa ₃₅ -Thr-Xaa ₃₇ ; Formula (I) (SEQ ID No: 3) wherein Xaa _{1 is} deleted or independently selected from Ala, Cys, Glu, Gly, His, Arg , Ser and Lys; Xaa _{3 is} independently selected from the group consisting of Gly, His, Arg, Ser and Asn; Xaa _{17 is} independently selected from the group consisting of His, Arg, Lys and Val; Xaa _{18 is} independently selected from Arg, Lys and His; Xaa _{21 is} independent Is selected from the group consisting of Ala, Lys, Gln, Ser and Asn; Xaa _{22 is} independently selected from Glu, Gln, Ser, Thr and Asn; Xaa _{26 is} independently selected from Pro, Arg and Ile; Xaa _{31 is} independently selected from Ser, Glu , Asp and Asn; Xaa _{35 are} independently selected from the group consisting of His, Arg, Lys, Asp and Glu; Xaa _{37 is} independently selected from Pro and Tyr; and wherein the C-terminus can be derivatized as appropriate; and the restriction condition is that the multi-peptide is not Is N-α-(19-carboxynodecyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]-planin; the multi-peptide is in its case Even a substituent group having at least one amino acid residue.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog of formula (I): Xaa ₁ -Cys-Xaa ₃ -Thr-Ala-Thr-Cys -Ala-Thr-Gln-Arg-Leu-Ala-Glu-Phe-Leu-Xaa ₁₇ -Xaa ₁₈ -Ser-Ser-Xaa ₂₁ -Xaa ₂₂ -Phe-Gly-Pro-Xaa ₂₆ -Leu-Pro-Pro- Thr-Xaa ₃₁ -Val-Gly-Ser-Xaa ₃₅ -Thr-Xaa ₃₇ ; Formula (I) (SEQ ID No: 3) wherein (a) Xaa _{1 is} deleted or independently selected from Ala, Cys, Glu, Gly, His, Arg, Ser and Lys; Xaa _{3 are} independently selected from Gly, His, Arg, Ser and Asn; Xaa _{17 is} independently selected from the group consisting of His, Arg, Lys and Val; Xaa _{18 is} independently selected from Arg, Lys and His; Xaa _{21 is} independently selected from the group consisting of Ala, Lys, Gln, Ser and Asn; Xaa _{22 is} independently selected from Glu, Gln, Ser, Thr and Asn; Xaa _{26 is} independently selected from Pro, Arg and Ile; Xaa _{31 is} independently selected from Ser, Glu, Asp and Asn; Xaa _{35 are} independently selected from the group consisting of His, Arg, Lys, Asp and Glu; Xaa _{37 is} independently selected from Pro and Tyr; and wherein the C-terminus can be derivatized as appropriate; and the limitation is that The peptide is not N-α-(19-carboxynonanoyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; and (b) the analog is at pH 4 Having a solubility of about 100 μM or greater than 100 μM; the polypeptide optionally has at least one substituent attached to at least one of its amino acid residues.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog of formula (I): Xaa ₁ -Cys-Xaa ₃ -Thr-Ala-Thr-Cys -Ala-Thr-Gln-Arg-Leu-Ala-Glu-Phe-Leu-Xaa ₁₇ -Xaa ₁₈ -Ser-Ser-Xaa ₂₁ -Xaa ₂₂ -Phe-Gly-Pro-Xaa ₂₆ -Leu-Pro-Pro- Thr-Xaa ₃₁ -Val-Gly-Ser-Xaa ₃₅ -Thr-Xaa ₃₇ ; Formula (I) (SEQ ID No: 3) wherein (a) Xaa _{1 is} deleted or independently selected from Ala, Cys, Glu, Gly, His, Arg, Ser, and Lys; Xaa _{3 is} independently selected from the group consisting of Gly, His, Arg, Ser, and Asn; Xaa _{17 is} independently selected from the group consisting of His, Arg, Lys, and Val; and Xaa _{18 is} independently selected from the group consisting of Arg, Lys, and His; Xaa _{21 is} independently selected from the group consisting of Ala, Lys, Gln, Ser and Asn; Xaa _{22 is} independently selected from Glu, Gln, Ser, Thr and Asn; Xaa _{26 is} independently selected from Pro, Arg and Ile; Xaa _{31 is} independently selected from Ser, Glu, Asp and Asn; Xaa _{35 are} independently selected from the group consisting of His, Arg, Lys, Asp and Glu; Xaa _{37 is} independently selected from Pro and Tyr; and wherein the C-terminus can be derivatized as appropriate; and the limitation is that The peptide is not N-α-(19-carboxynonanoyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; and (b) the analog is at pH 7. Greater than about 100 μM or 100 μM of solubility; the multi-peptide in which optionally at least one amino acid residue having at least one substituent attached.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog of formula (I): Xaa ₁ -Cys-Xaa ₃ -Thr-Ala-Thr-Cys -Ala-Thr-Gln-Arg-Leu-Ala-Glu-Phe-Leu-Xaa ₁₇ -Xaa ₁₈ -Ser-Ser-Xaa ₂₁ -Xaa ₂₂ -Phe-Gly-Pro-Xaa ₂₆ -Leu-Pro-Pro- Thr-Xaa ₃₁ -Val-Gly-Ser-Xaa ₃₅ -Thr-Xaa ₃₇ ; Formula (I) (SEQ ID No: 3) wherein (a) Xaa _{1 is} deleted or independently selected from Ala, Cys, Glu, Gly, His, Arg, Ser, and Lys; Xaa _{3 is} independently selected from the group consisting of Gly, His, Arg, Ser, and Asn; Xaa _{17 is} independently selected from the group consisting of His, Arg, Lys, and Val; and Xaa _{18 is} independently selected from the group consisting of Arg, Lys, and His; Xaa _{21 is} independently selected from the group consisting of Ala, Lys, Gln, Ser and Asn; Xaa _{22 is} independently selected from Glu, Gln, Ser, Thr and Asn; Xaa _{26 is} independently selected from Pro, Arg and Ile; Xaa _{31 is} independently selected from Ser, Glu, Asp and Asn; Xaa _{35 are} independently selected from the group consisting of His, Arg, Lys, Asp and Glu; Xaa _{37 is} independently selected from Pro and Tyr; and wherein the C-terminus can be derivatized as appropriate; and the limitation is that The peptide is not N-α-(19-carboxynonanoyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; and (b) the analog is at pH 4 Having a solubility of about 100 μM or greater; (c) the analog has a solubility of about 100 μM or greater at pH 7; the polypeptide is optionally attached to at least one of its amino acid residues At least one substituent.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog of formula (I): Xaa ₁ -Cys-Xaa ₃ -Thr-Ala-Thr-Cys -Ala-Thr-Gln-Arg-Leu-Ala-Glu-Phe-Leu-Xaa ₁₇ -Xaa ₁₈ -Ser-Ser-Xaa ₂₁ -Xaa ₂₂ -Phe-Gly-Pro-Xaa ₂₆ -Leu-Pro-Pro- Thr-Xaa ₃₁ -Val-Gly-Ser-Xaa ₃₅ -Thr-Xaa ₃₇ ; Formula (I) (SEQ ID No: 3) wherein (a) Xaa _{1 is} deleted or independently selected from Ala, Cys, Glu, Gly, His, Arg, Ser, and Lys; Xaa _{3 is} independently selected from the group consisting of Gly, His, Arg, Ser, and Asn; Xaa _{17 is} independently selected from the group consisting of His, Arg, Lys, and Val; and Xaa _{18 is} independently selected from the group consisting of Arg, Lys, and His; Xaa _{21 is} independently selected from the group consisting of Ala, Lys, Gln, Ser and Asn; Xaa _{22 is} independently selected from Glu, Gln, Ser, Thr and Asn; Xaa _{26 is} independently selected from Pro, Arg and Ile; Xaa _{31 is} independently selected from Ser, Glu, Asp and Asn; Xaa _{35 are} independently selected from the group consisting of His, Arg, Lys, Asp and Glu; Xaa _{37 is} independently selected from Pro and Tyr; and wherein the C-terminus can be derivatized as appropriate; and the limitation is that The peptide is not N-α-(19-carboxynonanoyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; and (b) the analog is at pH 4 Having a solubility of about 100 μM or greater; (c) the analog has a solubility of about 100 μM or greater than 100 μM at pH 7; (d) the multipeptide has about 25 hours or more in the fibril formation assay 25 hours of physical stability; the multipeptide is optionally attached to at least one of its at least one amino acid residue.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The analog comprises a histidine or arginine residue at position 17; (c) the analog comprises a histidine residue at position 35; wherein the analog amino acid sequence number of the analog corresponds to the SEQ ID No: 2 amino acid numbering sequence; and (d) the multi-peptide has a solubility of about 100 μM or more at pH 4; and the multi-peptide is optionally on at least one amino acid residue thereof There are at least one substituent attached.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The analog comprises a histidine or arginine residue at position 17; (c) the analog comprises a histidine residue at position 35; wherein the analog amino acid sequence number of the analog corresponds to the SEQ ID No: 2 amino acid numbering sequence; and (d) the multi-peptide has about 100 μM or more than 100 μM at pH 7. Solubility; and the multipeptide is optionally attached to at least one of its at least one amino acid residue.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The analog comprises a histidine or arginine residue at position 17; (c) the analog comprises a histidine residue at position 35; wherein the analog amino acid sequence number of the analog corresponds to the SEQ ID No: 2 amino acid numbering sequence; and (d) the multi-peptide has a solubility of about 100 μM or greater at pH 4; and (e) the multi-peptide has about 100 μM at pH 7 or A solubility greater than 100 μM; and the multipeptide is optionally attached to at least one of its at least one amino acid residue.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The analog comprises a histidine or arginine residue at position 17; (c) the analog comprises a histidine residue at position 35; wherein the analog amino acid sequence number of the analog corresponds to the SEQ ID No: 2 amino acid numbering sequence; and (d) the polypeptide peptide has about 25 hours or more in the fibril formation assay Physical stability over 25 hours; the multi-peptide is optionally attached to at least one of its at least one amino acid residue.

In another aspect, the invention relates to a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog, wherein: (a) the analog comprises a glutamic acid residue at position 14; The analog comprises a histidine or arginine residue at position 17; (c) the analog comprises a histidine residue at position 35; wherein the analog amino acid sequence number of the analog corresponds to the SEQ ID No: 2 amino acid numbering sequence; and (d) the multi-peptide has a solubility of about 100 μM or greater at pH 4; and (e) the multi-peptide has about 100 μM at pH 7 or a solubility greater than 100 μM; and (f) the multi-peptide has a physical stability of about 25 hours or more than 25 hours in the fibril formation assay; the multi-peptide is optionally attached to at least one of its amino acid residues At least one substituent.

Some advantages

The multi-peptide of the present invention can exhibit improved physical stability.

The multi-peptide of the present invention can exhibit improved solubility.

The multi-peptide of the present invention can exhibit improved physical stability and solubility.

Some preferred aspects

Assays are provided herein for measuring human amyloid receptor binding and potency, solubility, and physical stability (for example, see assays (II), (IV), and (III), respectively).

Xaa _{17 is} preferably His or Arg; more preferably Arg.

Xaa _{37 is} preferably Pro.

Xaa _{35 is} preferably His; Xaa _{17 is} preferably His and Xaa _{37 is} preferably Pro.

Xaa _{17 is} preferably Arg and Xaa _{37 is} preferably Pro.

Xaa _{17 is} preferably His and Xaa _{35 is} preferably His.

Xaa _{17 is} preferably Arg and Xaa _{35 is} preferably His.

In a preferred embodiment of the invention, a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog of formula (I), wherein: Xaa _{1 is} deleted or independently selected from the group consisting of His, Arg and Lys Xaa _{3 is} independently selected from the group consisting of Gly, His, and Asn; Xaa _{17 is} independently selected from the group consisting of His, Arg, and Val; Xaa _{18 is} independently selected from Arg and His; Xaa _{21 is} independently selected from Ser and Asn; and Xaa ₂₂ is Asn; Xaa ₂₆ is Ile; Xaa _{31 is} independently selected from Glu and Asn; Xaa _{35 is} independently selected from the group consisting of His, Arg, Lys, Asp, and Glu; and Xaa _{37 is} independently selected from Pro and Tyr.

In a preferred embodiment of the invention, a polypeptide comprising an amino acid sequence of the SEQ ID No: 2 analog of formula (I) wherein: Xaa _{1 is} deleted or independently selected from Gly, His, Arg , Ser and Lys; Xaa _{3 is} independently selected from the group consisting of His and Asn; Xaa _{17 is} independently selected from the group consisting of His, Arg and Val; Xaa _{18 is} independently selected from Arg and His; Xaa _{21 is} independently selected from Gln, Ser and Asn; Xaa ₂₂ is Asn; Xaa _{26 is} independently selected from Pro and Ile; Xaa ₃₁ is Asn; Xaa ₃₅ is His; and Xaa ₃₇ is Tyr.

In a preferred embodiment of the invention, a polypeptide comprising an amino acid sequence of the SEQ ID No: 2 analog of formula (I) wherein Xaa _{1 is} deleted or independently selected from Ala, Cys, Glu , Gly, His, Arg, Ser, and Lys; Xaa ₃ is Asn; Xaa _{17 is} independently selected from His, Arg, and Val; Xaa _{18 is} independently selected from Arg, Lys, and His; Xaa _{21 is} independently selected from Gln and Asn; Xaa _{22 is} independently selected from Thr and Asn; Xaa ₂₆ is Ile; Xaa ₃₁ is Asn; Xaa _{35 is} independently selected from Gly and Asn; and Xaa ₃₇ is Pro.

In a preferred embodiment of the invention, a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog of formula (I) wherein: Xaa _{1 is} deleted; Xaa _{3 is} independently selected from Gly and Asn Xaa _{17 is} independently selected from the group consisting of His, Arg and Val; Xaa _{18 is} independently selected from Arg and His; Xaa _{21 is} independently selected from Gln and Asn; Xaa _{22 is} independently selected from Gln and Asn; Xaa ₂₆ is Ile; Xaa ₃₁ Independently selected from Glu and Asn; Xaa _{35 is} independently selected from His and Ser; Xaa _{37 is} independently selected from Pro and Tyr.

In a preferred embodiment of the invention, a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog of formula (I) wherein: Xaa ₁ is Lys; Xaa ₃ is Asn; Xaa _{17 is} independent Is selected from the group consisting of His, Lys, Arg and Val; Xaa _{18 is} independently selected from Arg and His; Xaa _{21 is} independently selected from Ala, Lys, Gln and Ser; Xaa _{22 is} independently selected from Glu, Gln, Ser, Thr and Asn Xaa _{26 is} independently selected from the group consisting of Pro and Ile; Xaa _{31 is} independently selected from the group consisting of Ser, Glu, Asp, and Asn; Xaa _{35 is} independently selected from the group consisting of His, Glu, and Asn; and Xaa _{37 is} independently selected from Pro and Tyr.

Xaa _{37 is} independently selected from Pro and Tyr.

In a preferred embodiment of the invention, a multi-peptide comprising an amino acid sequence of the SEQ ID No: 2 analog of formula (I) wherein: Xaa ₁ is Lys; Xaa ₃ is Asn; Xaa ₁₇ is His or Arg; Xaa ₁₈ is His; Xaa ₂₁ is Asn; Xaa ₂₂ is Asn; Xaa ₂₆ is Ile; Xaa ₃₁ is Asn; Xaa ₃₅ is His or Asn; and Xaa ₃₇ is Pro.

In a specific example, the C-terminus can be derivatized.

In one embodiment, the C-terminus of the multi-peptide can be blocked with an acid or a guanamine. In one aspect, the C-terminus of the multi-peptide is decylamine.

In one embodiment, the C-terminus is derived from a guanamine of formula (II): (II) C(O)NR ¹ R ²

Wherein R ¹ and R ^{2 are} independently selected from H and an alkyl group. R ¹ and R ^{2 are} preferably both H.

In one embodiment, the multi-peptide of the present invention may have a substituent attached to any available position on one or more amino acid residues. Examples of substituents include a chemical moiety that is directly bonded to one or more amino acid residues, or an indirect moiety that binds to one or more amino acid residues by means of a linker. The available connection points are known to those skilled in the art. Examples of available connection points include multiple N-terminal of peptide; C-terminal of polypeptide; ε-amine group of amino acid residue; hydroxyl group of residue of serine, tyrosine or threonine; aspartic acid or glutamic acid residue Amine amino group; a carboxyl group of aspartic acid or a glutamic acid residue; a thiol group of a cysteine residue. The substituent is preferably attached to the N-terminus of the polypeptide or to the epsilon-amine group of the amine acid residue.

In another embodiment, the substituent is attached to the N-terminal amine group of the multi-peptide, wherein the N-terminal amino acid residue corresponds to position 1 of the SEQ ID No: 2 analog.

In another embodiment, the substituent is attached to the epsilon-amine group of the amine acid residue in position 1 of the analog of SEQ ID No: 2.

In one embodiment, the substituent is selected from the group consisting of a hydrocarbyl substituent, a hydroxyl group, and a halogen atom. Examples of suitable halogen atoms include F, Cl, Br, and I. The substituent is preferably a hydrocarbyl substituent.

In another embodiment, the hydrocarbyl substituent is an alkyl group or a group of formula (III): (III) L _n -Y

Where L is a linker; n=0 or 1

Y is a chemical moiety such as an albumin binding moiety.

In one embodiment, the linker comprises from 1 to 10 amino acids. The linker may further comprise an amine.

Examples of suitable amines include: -C(O)-(CH ₂ ) _l -O-[CH ₂ CH ₂ -O] _m -(CH ₂ ) _p [NHC(O)-(CH ₂ ) _l O- [( CH ₂ ) _n -O] _m -(CH ₂ ) _p ] _q -NH-

Wherein l, m, n and p are independently 1-7, and q is 0-5.

For example, the linker can comprise an amine selected from the group consisting of: -C(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -NH-; and -C(O)-CH _2- O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -[NHC(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -] ₁ -NH-; -C(O)-(CH ₂ ) ₂ -O-[CH ₂ CH ₂ -O] ₇ -(CH ₂ ) ₂ -NH-.

In another embodiment, the linker is a combination of an amino acid residue and the above amine, for example: γGlu-C(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -[ NHC(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -] ₁ -NH-; or Arg-Arg-γGlu-C(O)-CH ₂ -O-CH ₂ - CH ₂ -O-CH ₂ -CH ₂ -[NHC(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -] ₁ -NH-.

In some embodiments, n=1 and L are selected from the group consisting of γGlu, γGlu-γGlu, γGlu-γGlu-γGlu, γGlu-γGlu-γGlu-γGlu, Glu, Glu-Glu, Glu-γGlu, Glu- Arg, Glu-Glu-Arg, His, His-His, His-γGlu, His-His-γGlu, Gly, Gly-γGlu, Ser, Ser-γGlu, D-Arg-D-Arg, Arg, Arg-Arg, Arg-Arg-γGlu, Ser-Ser, -Gly-Ser-Ser, Ser-Ser, -Gly-Ser-Ser-γGlu, Ser-Ser-Gly-Ser-Ser-Gly and Ser-Ser-Gly-Ser- Ser-Gly-γGlu, γGlu-OEG, γGlu-OEG-OEG and OEG.

In some embodiments, n=1 and L are selected from the group consisting of γGlu, γGlu-γGlu, γGlu-OEG, γGlu-OEG-OEG, and OEG, and the linker is more preferably γGlu-γGlu.

In another embodiment, n = 0; thus there is no linker between the amino acid residue of the polypeptide backbone and the chemical moiety Y, i.e., Y is attached to the available position on the polypeptide backbone.

In a specific example, Y is an albumin binding moiety.

In a specific example, the albumin binding moiety is a sulfhydryl group.

The albumin binding moiety is preferably HOOC(CH ₂ ) _s CO-, wherein s is an integer from 12 to 22. s is more preferably an integer from 14 to 20, such as 14, 15, 16, 17, 18, 19 or 20. s is preferably 16 to 18. s is better at 18.

In a specific embodiment, the multi-peptide of the present invention comprises a γGlu linker linked to the N-terminus of the amyloid analog and HOOC(CH ₂ ) ₁₈ CO or HOOC(CH ₂ ) ₁₆ CO- as an albumin-binding residue, And wherein the sequence of the amyloid analog comprises a Glu at position 14, a His or Arg at position 17, a His at position 35 or a Pro at position 37, as compared to SEQ ID NO: 2.

In another embodiment, the substituent and/or the group of formula (III) is selected from the group consisting of the groups provided in Table 1.

The multipeptide of the present invention preferably comprises a substituent.

Preferably, the multi-peptide of the present invention has a substituent attached to the N-terminal amine group of the multi-peptide, wherein the N-terminal amino acid residue corresponds to position 1 of the analog of SEQ ID No: 2.

The substituent is preferably:

Substituent:

Preferably, it is attached to the N-terminal amino acid group of the multi-peptide.

For specific examples comprising albumin binding moieties, the multi-peptides of the invention may exhibit extended pharmacokinetic characteristics and good pharmacodynamic properties. Therefore, the multi-peptide of the present invention does not have to be injected as frequently as the known amylin product.

Furthermore, the multipeptide of the present invention reduces food intake. The food intake is better than the known amyloid product.

In a specific example, the albumin binding moiety binds non-covalently to albumin. The albumin binding moiety preferably has an albumin binding affinity for human serum albumin of less than about 10 μM or less than about 1 μM.

In one embodiment, the multi-peptide of the invention is selected from the following compounds provided in Table 2 (below). Table 2 provides a list of compounds having a solubility greater than 100 μM at pH 4.

As demonstrated in the Examples section herein, the multi-peptide provided above has a solubility greater than or equal to 100 μM (micro-mole concentration) at pH 4.

In one embodiment, the multipeptide of the invention has a solubility of greater than or equal to 125 μM (micromolar concentration) at pH 4.

In one embodiment, the multipeptide of the invention has a solubility of greater than or equal to 150 μM (micromolar concentration) at pH 4.

In one embodiment, the multipeptide of the invention has a solubility of greater than or equal to 175 μM (micromolar concentration) at pH 4.

In one embodiment, the multipeptide of the invention has a solubility of greater than or equal to 200 μM (micromolar concentration) at pH 4.

In one embodiment, the solubility is measured in a solubility assay as provided herein.

In another embodiment, the multi-peptide of the invention is selected from the following compounds provided in Table 3 (below).

Table 3 provides a list of compounds having a solubility greater than 100 μM at pH 7.

As demonstrated in the Examples section herein, the multi-peptide provided above has a solubility greater than or equal to 100 μM (micro-mole concentration) at pH 7.

In a specific example, the multi-peptide of the invention has a greater than pH 7 Or equal to the solubility of 125 μM (micromolar concentration).

In one embodiment, the multipeptide of the invention has a solubility of greater than or equal to 150 μM (micromolar concentration) at pH 7.

In one embodiment, the multipeptide of the invention has a solubility of greater than or equal to 175 μM (micromolar concentration) at pH 7.

In one embodiment, the multipeptide of the invention has a solubility of greater than or equal to 200 μM (micromolar concentration) at pH 7.

In one embodiment, the solubility is measured in a solubility assay as provided herein.

Table 4 provides a list of compounds having a solubility greater than 100 μM at both pH 4 and pH 7.

As demonstrated in the Examples section herein, the multi-peptide provided above has a solubility of greater than or equal to 100 μM (micromolar concentration) at both pH 4 and pH 7.

In one embodiment, the multipeptide of the present invention has a solubility of greater than or equal to 125 μM (micromolar concentration) at both pH 4 and pH 7.

In one embodiment, the multipeptide of the present invention has a solubility of greater than or equal to 150 μM (micromolar concentration) at both pH 4 and pH 7.

In one embodiment, the multipeptide of the present invention has a solubility of greater than or equal to 175 μM (micromolar concentration) at both pH 4 and pH 7.

In one embodiment, the multipeptide of the present invention has a solubility of greater than or equal to 200 μM (micromolar concentration) at both pH 4 and pH 7.

In one embodiment, the solubility is measured in a solubility assay as provided herein.

Table 5 provides a list of compounds having physical stability greater than or equal to 25 hours in the fibril formation assay.

As demonstrated in the Examples section herein, the multi-peptide provided above has a physical stability of greater than or equal to 25 hours in the fibril formation assay.

In one embodiment, the multipeptide of the invention has a physical stability of greater than or equal to 30 hours.

In one embodiment, the multipeptide of the invention has a physical stability of greater than or equal to 35 hours.

In one embodiment, the multipeptide of the invention has a physical stability of greater than or equal to 40 hours.

In one embodiment, the multipeptide of the invention has a physical stability of greater than or equal to 45 hours.

In one embodiment, the physical stability is measured in a fibril formation assay as provided herein.

Table 6 provides with 1800 pM (picomolar concentrations), or less than 1800 pM of human receptor starch compounds value list ₅₀ EC.

As demonstrated in the Examples section herein embodiments, provide much above peptide having 1800 pM (picomolar concentration) value of ₅₀ or less than human amylin of EC 1800 pM.

In a specific example, the present invention has many peptide 1500 pM (picomolar concentration) value of ₅₀ or less than human amylin of EC 1500 pM.

In a specific example, the present invention has many peptide 1200 pM (picomolar concentration) value of ₅₀ or less than human amylin of EC 1200 pM.

In one embodiment, the present invention is much peptide having 1000 pM (picomolar concentration) value of ₅₀ or less than human amylin of EC 1000 pM.

In a specific example, the present invention has many peptide 800 pM (picomolar concentration) value of ₅₀ or less than human amylin of EC 800 pM.

In a specific example, the present invention has many peptide 600 pM (picomolar concentration) value of ₅₀ or less than human amylin of EC 600 pM.

In a specific example, the present invention has many peptide 400 pM (picomolar) or a value less than ₅₀ human amylin of EC 400 pM.

In one embodiment, the present invention is much peptide having 200 pM (picomolar concentrations), or less than human amylin of ₅₀ EC 200 pM value.

In a specific example, the present invention has many peptide 100 pM (picomolar concentration) value of ₅₀ or less than human amylin of EC 100 pM.

In a specific example, the present invention has many peptide 75 pM (picomolar concentration) value of ₅₀ or less than human amylin of EC 75 pM.

In a specific example, the present invention has many peptide 60 pM (picomolar concentration) value of ₅₀ or less than human amylin of EC 60 pM.

In a specific example, in the assay as herein provided in the measuring human amylin EC _50.

Table 7 provides with 1800 pM (picomolar concentration) list of compounds, or less than 1800 pM of human amylin receptor the EC ₅₀ value.

As demonstrated in the Examples section herein embodiments, provide much above peptide having 1800 pM (picomolar concentrations), or less than human calcitonin EC ₅₀ value of 1800 pM.

In a specific example, the present invention has many peptide 1500 pM (picomolar concentration) value of ₅₀ or less than human calcitonin of EC 1500 pM.

In a specific example, the present invention has many peptide 1200 pM (picomolar concentration) value of ₅₀ or less than human calcitonin of EC 1200 pM.

In one embodiment, the present invention is much peptide having 1000 pM (picomolar concentrations), or less than human calcitonin EC ₅₀ value of 1000 pM.

In a specific example, the present invention has many peptide 800 pM (picomolar concentrations), or less than human calcitonin EC ₅₀ value of 800 pM.

In a specific example, the present invention has many peptide 600 pM (picomolar concentration) value of ₅₀ or less than human calcitonin the EC 600 pM.

In a specific example, the present invention has many peptide 400 pM (picomolar concentrations), or less than human calcitonin EC ₅₀ value of 400 pM.

In one embodiment, the present invention is much peptide having 200 pM (picomolar concentrations), or less than human calcitonin EC ₅₀ value of 200 pM.

In a specific example, the present invention has many peptide 100 pM (picomolar concentrations), or a value less than ₅₀ human calcitonin The EC 100 pM.

In a specific example, the present invention has many peptide 80 pM (picomolar concentrations), or less than ₅₀ human calcitonin values of EC 80 pM.

In a specific example, in the assay as herein provided in the measuring human calcitonin EC _50.

Table 8 provides with 1800 pM (picomolar), or less than 1800 pM of human receptor starch ₅₀ values EC, 150 μM or more and a solubility of 25 hours in assay formed fibrils at pH 150 μM 7 25 or more hours A list of compounds with physical stability.

In a specific example, the present invention has many peptide 1800 pM (picomolar concentrations), or a value less than ₅₀ human amylin of EC 1800 pM and 150 μM is greater than the solubility at pH 7, and is formed in the fibril assay Physical stability greater than or equal to 25 hours.

In a specific example, in the assay as herein provided in the measuring human amylin EC _50, the solubility and physical stability of 7 in pH.

Table 9 provides with 1800 pM (picomolar concentrations), or less than 1800 pM of human amylin receptor ₅₀ values EC, solubility above pH 7 or above 200 μM and 200 μM of fibril formation assay at 45 hours or 45 hours A list of compounds with physical stability.

In a specific example, the present invention has many peptide 1800 pM (picomolar concentrations), or a value less than ₅₀ human amylin of EC 1800 pM and 200 μM is greater than the solubility at pH 7, and is formed in the fibril assay Physical stability greater than or equal to 45 hours.

In a specific example, in the assay as herein provided in the measuring human amylin EC _50, the solubility and physical stability of 7 in pH.

In one aspect of the invention, the amyloid derivative is selected from the group consisting of N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[His1 , Glu14, His17, Arg18]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[His1, Glu8, Glu14, His17, Arg18 ]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[des1, Gly3, Glu14, His17, Arg18, Glu31, Gly35]- Pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxyundecylamino)butanyl]-[des1, Gly3, Glu14, His17, Arg18, Gln21, Gln22, Glu31, Gly35]-Pullin Peptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu1, Glu14, His17, Arg18]-planin, N-α-[( S)-4-carboxy-4-(19-carboxyundosylamino)butanyl]-[His1, Glu14, His17, Arg18, Ala25, Pro26, Ser28, Ser29]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxyundosylamino)butanyl]-[His1, Glu14, His17, Arg18, Glu25, Ser28, Arg29]-pramlintide, N-α-[(S) 4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1, Glu14, His17, Arg18, Glu22, Ala25, Arg26, Ser28, Ser29]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1, Glu14, His17, Arg18, Ser21, Ser22, Asp31, Asp35]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxyundosylamino)butanyl]-[Glu1, Glu14, His17, Arg18, Pro37]-pramlintide, N-α-[(S)-4-carboxyl -4-(19-carboxynonylamino)butanyl]-His-His-[His1,Glu14,Hi17]-pramlintide, N-α-[(S)-4-carboxy-4-(19- Carboxylamidylamino)butanyl]-His-His-[His1, Glu14, Arg17 ]-pramlintide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxy) Amidino) butylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Arg1, Glu14, His17, Arg18, Lys21]-pramlintide , N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1,Glu14,Arg18]-planin, N-α-[(S)- 4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1, Gly3, Glu14, His17, Arg18, Ser21, Ser22, Glu31, Glu35]-pramlintide, N-α-[(S )-4-carboxy-4-(19-carboxynonylamido)butanyl]-His-[His1,Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4- (19-carboxynonylamido)butanyl]-[His1,Glu14,Hi17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)醯醯基]-His-His-[His1, Glu14, His17, Ala25, Pro26, Ser28, Ser29]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonadecanamide Butyl)-His-His-[His1,Glu14]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-His-His -[His1,Glu14,His17,Arg35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-His-His-[His1, Arg3, Glu14, His17]-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-) 4-(19-carboxyheptadecylamido)butanylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[His1,Glu14, His17, Arg18, Ala19, Thr20, Gln21, Glu22, Leu23]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu1, Gly3, Glu14, Arg18, Ser21, Ser22, Ala25, Pro26, Ser28, Ser29, Glu31 , Arg35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-[Arg1, Glu14, His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-[Glu14,His17]-pramlintide, N-α-[(S)-4 -carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxy-19醯Amino)butanyl]-[des1,Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu1, Glu14 ,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu1, Glu14, His17, Arg18]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-His-His-[His1, Glu14, His17, His35]-pramlintide, N-α- [(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-His-His-[His1,His3,Glu14,His17,His35]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-His-His-[His1,His3,Glu14 ,His17,His29,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonanosylamino)butanyl]-His-His-[His1, Glu14, His17 ,His29,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-His-His-[His1,His3,Glu14,His17]-pramlintide, N-α- [(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-His-His-[His1,Glu14,His17,His29]-pramlintide, N-α-[2-( 2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadecylamido)butaninyl]ethoxy}ethoxy)acetamidine Amino]ethoxy}ethoxy)ethinyl]-D-Arg-D-Arg-[Glu14,His17,Arg18]-pramlintide, N-α-[2-(2-{2-[ 2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadecanosamino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy }Ethoxy)ethinyl]-D-Arg-D-Arg-[Glu14,His17,Arg18,Ala21,Ser35]-pramlintide, N-α-[(S)-4-carboxy-4-( 19-carboxy-nonyl-nonylamino)butanyl]-Ser-Ser-Gly-Ser-Ser-[Gly1, Glu14, His17]-pramlintide, N-α-[(S)-4-carboxy-4-( 19-carboxynonanosylamino)butanyl]-Glu-[His1,Glu14,Hi17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)醯Germanyl]-Glu-Glu-[Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-Glu-[Glu14 ,His17]-pramlintide, N-α- [2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadelamido))butanylamino]ethoxy}ethoxy Ethylamino]ethoxy}ethoxy)ethinyl]-[des1,Glu14,Hi17]-pramlintide, N-α-[(S)-4-carboxy-4-(19- Carboxylamidylamino)butanyl]-Ser-Ser-Gly-Ser-Ser-Gly-[His1,Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-His-His-[His1,Glu14,His17]-pramlintide, N-α-[( S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-[Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(17- Carboxythylideneamino)butanyl]-His-[His1,Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptylamino)butanyl ]-[His1,Glu14,His17]-pramlintide, N-α-{(S)-4-carboxy-4-[(S)-4-carboxy-4-(17-carboxyheptadecaneamino) Butylamine]butyrate}-[des1,Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14 ,Arg17]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-[Glu14,Arg17]-pramlintide, N-α- [(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14,His17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4 -(19-carboxynonylamido)butanyl]-[Glu14,Arg17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino) ) 醯 醯]]-[Glu10, Glu14, His17]- pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxyundecylamino) butyl -[Glu14,His17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-[Glul4,His17, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14,Arg17,His35]-pramlintide, N-α-[(S)-4 -carboxy-4-(19-carboxyundecylamino)butanyl]-Glu-[Glu14, Arg17, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(17- Carboxythylideneamino)butanyl]-[Glu14,His17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptylamino)butanyl]- Glu-[Glu14,His17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-[Glu14, Arg17, His35]- Pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptadelamido)butanyl]-Glu-[Glu14, Arg17, His35]-pramlintide, N-α- [(S)-4-carboxy-4-(19-carboxyundosylamino)butanyl]-His-His-[His1, Glu14, His17, Ser21, Glu35]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14,His17,Ala25,Pro26,Ser28,Ser29,His35]-pramlintide, N-α-[(S) 4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14, Arg17, Ala25, Pro26, Ser28, Ser29, His35]-pramlintide, N-α-[(S)-4 -carboxy-4-(19-carboxyundecylamino)butanyl]-His-[Glu14,His17,His35]- LAN Lin peptide, N-α - [(S) -4- Carboxy-4- (19-carboxynonadecanoylamino acyl amino) butan-acyl] -His-His- [Glu14, His17, His35] - pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-His-[Glu14, Arg17, His35]-pramlintide, N-α-[(S) 4-carboxy-4-(19-carboxynonylamino)butanyl]-His-His-[Glu14, Arg17, His35]-pramlintide, N-α-[(S)-4-carboxy-4 -(17-carboxyheptadecaneamino)butanyl]-His-[Glu14,His17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyl-17醯Amino)butanyl]-His-[Glu14, Arg17, His35]-pramlintide, N-ε-25-[2-(2-{2-[2-(2-{2-[(S)-4) -carboxy-4-(17-carboxyheptadecylamido)butanylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Ser21 , Lys25]-pramlintide, N-ε-25-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxy] Amino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Lys25]-planin, N-ε- 21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanium))butanylamino]ethoxy} Ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Lys21,Glu35]-pramlintide, N-α-[(S)-4-carboxy-4 -(19-carboxynonylamino)butanyl]-[des 1, Glu14, His17, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[His1, Glu14, His17, His35] - pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1, Glu14, His17, Arg18, His35]-pramlintide, N -α-[(S)-4-carboxy-4-(19-carboxyundecylamino)butane -[Glu14,His17,Arg18,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14,His17, Arg18]-pramlintide, N-ε-17-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecane) Butylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,Lys17]-planin, N-α-[(S )-4-carboxy-4-(19-carboxynonylamido)butanyl]-Gly-[Glu14,His17,His35]-planin, N-α-[2-(2-{2-[2 -(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanosamino)butaninyl]ethoxy}ethoxy)ethinyl]ethoxy} Ethoxy)ethinyl]-[Glu1, Glu14,His17,Pro37]-pramlintide, N-ε-17-[2-(2-{2-[2-(2-{2-[(S) )-4-carboxy-4-(17-carboxyheptadecylamido)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[ Glu14, Lys17, Ser21, Glu35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[His1, Glu14, His17, Pro37] - pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Ser-[Glu14,His17,His35]-pramlintide, N-α -[(S)-4-carboxy-4-(19- N-decylamino)butanyl]-[Glu14,Pro37]-pramlintide, N-ε-1-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]- [Glu14, His17, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Gly1, Glu14, Arg17, Pro37]- Pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Gly-[His1,Glu14,Arg17,Pro37]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1,Glu14,Arg17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4 -(19-carboxynonylamino)butanyl]-[Gly1, Glu14, His17, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxyl-19醯Amino)butyric acid]-Gly-[His1,Glu14,His17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]- [Glu14, His35]-pramlintide, N-ε-1-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-[Glu14,His17,His35]-Planin Peptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Arg1, Glu14, His17, His35]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Arg1,Glu14,His17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4 -(19-carboxynonylamino)butanyl]-[Arg1,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl ]-[Arg1,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butyl Mercapto]-Gly-[Arg1,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Gly-[Arg1, Glu14 ,His17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxyundecylamino)butanyl]-Gly-[Arg1, Glu14, His17, Pro37]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxyundecylamino)butanyl]-Gly-[Arg1,Glu14,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4 -(19-carboxynonylamido)butanyl]-Gly-[Arg1,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino) )]][[des1,Glu14,His17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Gly1,Glu14 ,His17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Ser1, Glu14, His17, Pro37]-Planin Peptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Ser1, Glu14, His17, His35]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[des1,Glu14,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-( 19-carboxynonylideneamino)butanyl]-[His1,Glu14,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl ]-[His1, Glu14, Ser21, Ser28, Ser29, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19- N-nonylamino)butanyl]-[His1, Glu14, Arg17, Ser21, Ser28, Ser29, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxyl Amidoxime -[Glu14,His17,Arg35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14,His17,Lys35] - pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14,His17,His36]-pramlintide, N-α-[ (S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14,His17,His34]-pramlintide, N-α-[(S)-4-carboxy-4- (19-carboxynonylamido)butanyl]-[Glu14,His17,His32]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)醯醯基]-[Glu14,His17,Gln21,His35)-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-) (19-carboxyheptadecylamido)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Ser21,His35] -Pullinin, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptylamino))) Amino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[His1, Glu14, His17, Ser21, His35]-pramlintide, N-α-[ 2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadelamido))butanylamino]ethoxy}ethoxy Ethylamine ethoxylate }Ethoxy)ethinyl]-D-Arg-D-Arg-[Orn1,Glu14,His17,Arg18]-pramlintide, N-α-(19-carboxyundecyl)-Glu-Glu- Arg-[Glu1,Pro37]-pramlintide N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-Arg-[Glu1,Pro37]-Plan Linpeptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Glu-Arg-[Glu1, Glu14, His17, Pro37]-pramlintide, N-α- [(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Glu-Arg-[Glu1, Glu14, Arg17, Pro37]-pramlintide, N-α-(19-carboxyl 19-mercapto)-Glu-Glu-Arg-[Glu1, Glu14, His17, Pro37]-pramlintide, N-ε-25-[2-(2-{2-[2-(2-{2- [(S)-4-carboxy-4-(17-carboxyheptadecylamido)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethenyl ]-[Glu14,Lys25]-pramlintide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-) Carboxyrytylamino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Lys21, Ser29, His35]-Planin Peptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, His17, Asp21, His35]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Glu-[Glu14,His17,Gln21,His35]-pramlintide, N-α-[(S)-4-carboxyl -4-(19-carboxynonylamido)butanyl]-[Glu14,His17,Gln21,Pro37]-pramlintide, N-ε-21-[2-(2-{2-[2-(2) -{2-[(S)-4-carboxy-4-(19- Carboxythylidene amino) butylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Lys21, Ser28, Ser29, His35]- Pramlintide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptylamino)) Amidino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Lys21,His35]-pramlintide, N-ε-25-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptylamino)butanylamino]] Ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Ser21,Lys25,Ser28,Ser29.His35]-pramlintide, N-ε- 25-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptytylamino)butanylamino]ethoxy} Ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[His17,His21,Lys25,Ser28,Ser29.His35]-pramlintide, N-α-[(S)-4 -carboxy-4-(19-carboxyundecylamino)butanyl]-[Glu14,His17,Gln35,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19- Carboxylamidinyl)butanyl]-[Glu14,His17,Gln21,Gln35,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino) )]][Gly1, Glu14, His17, Gln21, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]- [Glu1, Glu14, Gln21, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, Gln21, Gln35 ,Pro37]-pramlintide, N-ε-25-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxy] Amino)butynylamino] Ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[His17, Lys25, Ser28, Ser29, His35]-pramlintide, N-ε-21-[2 -(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptylamino)butaninyl]ethoxy}ethoxy) Ethylamino]ethoxy}ethoxy)ethinyl]-[His17,Lys21,Ser28,His35]-pramlintide, N-ε-25-[2-(2-{2-[2- (2-{2-[(S)-4-carboxy-4-(19-carboxy seventeen) Amidino) butylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Lys25,Ser28,Ser29,His35]-Plan Linpeptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1,Glu14,Arg18,Pro37]-pramlintide, N-ε-21 -[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptytylamino)butaninyl]ethoxy}B Oxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Lys21,Ser28,His35]-pramlintide, N-α-[(S)-4-carboxy- 4-(19-carboxynonylamido)butanyl]-[Lys11,Glu14,His17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxy-19 Amidino) butyl sulfhydryl]-[Glu14,His17,Thr21,Pro37]-pramlintide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-) 4-carboxy-4-(19-carboxyheptadecylamido)butanylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Lys21, Ser28, Arg29, His35]-pramlintide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19) -carboxyheptadecaneamino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Lys21,Ser28,Ser29, His3 5]-pramlintide, N-ε-1-[(S)-4-carboxy-4-(19-carboxynonanosylamino)butanyl]-Arg-[Glu14, Arg17, His35, Pro37]-Plan Lin peptide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadecanoyl))) Amino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Arg18, Lys21, Ser28, Arg29, Arg35]-pramlintide, N-ε -1-[(S)-4-carboxy-4-(19-carboxyundecylamino)butane Base]-[Glu14,His17,Gln21,Ala25,Pro26,Ser28,Ser29,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino) Butyl]-[Arg14, Arg17, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Gly-[Arg1, Glu14, Arg17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Arg17,Pro37]-pramlintide, N-α -[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Glu-Glu-[Arg1,Glu14,Arg17,Pro37]-pramlintide, N-α-[(S )-4-carboxy-4-(19-carboxyundecylamino)butanyl]-Glu-[Arg1,Glu14,Arg17,Pro37]-pramlintide, N-α-[(S)-4-carboxy- 4-(19-carboxynonylamido)butanyl]-Glu-Glu-[Arg1, Glu14, Arg17, Gln21, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4- (19-carboxynonylamido)butanyl]-Glu-Glu-[Arg1, Glu14, Arg17, Gln21, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4- (19-carboxynonylamido)butanyl]-Glu-Glu-[Arg1, Glu14, Arg17, Gln21, Gln31, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy- 4-(19-carboxynonylamido)butanyl]-Glu-Glu-[Arg1, Glu14, Arg17, Gl N21, Gln22, Gln31, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylideneamino)butane ]]-Glu-Arg-[Glu1, Glu14, Arg17, Gln21, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl] -Glu-Arg-[Glu1, Glu14, Arg17, Gln21, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl] -Glu-Arg-[Glu1, Glu14, Arg17, Gln21, Gln31, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino) Butyl]-Glu-Arg-[Glu1, Glu14, Arg17, Gln21, Gln22, Gln31, Gln35, Pro37]-Pullin, N-α-[(S)-4-carboxy-4-(19-carboxy-19 Amidino)butyric acid]-Glu-Glu-[Glu1, Glu14, Arg17, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)丁醯基]-[Glu14, Arg18, Gln21, Ala25, Pro26, Ser28, Ser29, Pro37]-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S) )-4-carboxy-4-(19-carboxyheptadecylamido)butanylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[ Glu14, His17, Glu21, Pro37]-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyl) Hexamethylamino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Ly S21,Pro37]-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadecane) Butylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Ala21,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14, Arg18, Ser21, Ser22, Ser28, Ser29, Asp31, Asp35, Pro37]-Pullin Peptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, Ser21, Ser22, Pro23, Ala25, Pro26, Ser28, Ser29, Asp31, Asp35 ,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14, Arg17, Asp21, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14, Arg17, Asp35, Pro37]-pramlintide, N-α-[(S) 4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14, Arg17, Asp31, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-( 19-carboxynonylamidoalkyl)butanyl]-[Glu14, Arg17, Asp22, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)醯]][[Asp3,Glu14,Arg17,Pro37]-pramlintide, N-ε-1-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14 ,Arg17,Pro37]-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxy] Amino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl] - [Glu14, Ala21, His35, Pro37] - pramlintide. For some specific examples, the multi-peptide of the present invention has prolonged pharmacokinetic characteristics compared to pramlintide as measured by assay (IX) as described in the assay section.

For some embodiments, the polypeptide peptides of the invention have a plasma T1⁄2 of at least 30 hours.

For some specific examples, the multi-peptide of the invention has a plasma T1⁄2 of at least 40 hours.

For some embodiments, the multipeptide of the invention has a plasma T1⁄2 of at least 50 hours.

For some specific examples, the multi-peptide of the invention has a plasma T1⁄2 of at least 60 hours.

For some specific examples, the multi-peptide of the invention has a plasma T1⁄2 of at least 70 hours.

For some specific examples, the multi-peptide of the invention has a plasma T1⁄2 of at least 75 hours.

For some embodiments, the multi-peptide of the invention has a plasma T1⁄2 of at least 80 hours.

For some embodiments, the multipeptide of the invention has a plasma T1⁄2 of at least 85 hours.

For some embodiments, the multipeptide of the invention has a plasma T1⁄2 of at least 90 hours.

For some embodiments, the multi-peptide of the invention has a plasma T1⁄2 of at least 95 hours.

For some embodiments, the multi-peptide of the invention has a plasma T1⁄2 of at least 100 hours.

method

Preparation of multi-peptides such as amyloid or its analogs is the technology Well known in the middle. The multi-peptides of the invention can thus be prepared by conventional multi-peptide synthesis, for example using t-Boc or Fmoc chemistry or other established techniques for solid phase multi-peptide synthesis, see for example Greene and Wuts, "Protective Groups in Organic Synthesis", John Wiley & Sons, 1999. The multi-peptide can also be produced by a method comprising the steps of: cultivating a host containing the DNA sequence encoding the multi-peptide and capable of expressing the multi-peptide, in a suitable nutrient medium, under conditions permitting expression of the multi-peptide cell. For multi-peptides comprising unnatural amino acid residues, the recombinant cells should be modified to incorporate the non-natural amino acid into the multi-peptide, for example by using tRNA mutants.

Pharmaceutical composition

In one aspect, the invention is directed to a pharmaceutical composition comprising a multi-peptide of the invention and a pharmaceutically acceptable excipient. These compositions are suitable for parenteral administration.

In one embodiment, the multipeptide is present in the formulation at a concentration of from about 0.1 mg/ml to about 25 mg/ml. In another aspect, the multipeptide is present in the formulation at a concentration of from about 1 mg/ml to about 10 mg/ml.

In another embodiment, the formulation has a pH of from 2.0 to 10.0. In another embodiment, the formulation has a pH of from 2.0 to 7.0. In another embodiment, the formulation has a pH of from 2.5 to 4.5. In another embodiment, the formulation has a pH of from 3.5 to 4.5.

The pharmaceutical composition containing the multi-peptide of the present invention can be prepared by a conventional technique such as that described in Remington's Pharmaceutical Sciences , 1985 or Remington: The Science and Practice of Pharmacy , 19th edition, 1995.

The formulation may further comprise a buffer system, a preservative, an isotonic agent, a chelating agent, a stabilizer, and/or a surfactant. The use of such excipients in pharmaceutical compositions is well known to those skilled in the art. For convenience, reference is made to Remington: The Science and Practice of Pharmacy , 19th edition, 1995.

In one embodiment, the pharmaceutical formulation is an aqueous formulation, that is, a formulation comprising water. The formulation is typically a solution or suspension. In another embodiment of the invention, the pharmaceutical formulation is an aqueous solution.

The term " aqueous formulation " is defined as a formulation comprising at least 50% w/w water.

Similarly, the term " aqueous solution " is defined as a solution comprising at least 50% w/w water, and the term " aqueous suspension " is defined as a suspension comprising at least 50% w/w water.

In another embodiment, the pharmaceutical formulation is a freeze-dried formulation to which a physician or patient adds a solvent and/or diluent prior to use. In another embodiment, the pharmaceutical formulation is a ready-to-use dry formulation (eg, freeze-dried or spray-dried) that does not require any prior dissolution.

" Dried form " means a liquid pharmaceutical composition or formulation by freeze drying (ie, lyophilization; see, eg, Williams and Polli (1984) J. Parenteral Sci. Technol. 38: 48-59), spray Drying (see Masters (1991) in Spray-Drying Handbook (5th ed.; Longman Scientific and Technical, Essez, UK), pp. 491-676; Broadhead et al. (1992) Drug Devel. Ind. Pharm. 18:1169- 1206; and Mumenthaler et al. (1994) Pharm. Res. 11: 12-20) or air dried (Carpenter and Crowe (1988) Cryobiology 25: 459-470; and Roser (1991) Biopharm. 4: 47-53).

In another embodiment of the invention, the buffering agent is selected from the group consisting of acetates, carbonates, citrates, glycine glycine, histidine, glycine, lysine, spermine Acid, dihydrogen phosphate, hydrogen phosphate, phosphate and ginseng (hydroxymethyl)-aminomethane, dihydroxyethylglycine, trishydroxyglycine, malic acid, lactic acid, succinate, cis Butenedioic acid, fumaric acid, tartaric acid, aspartic acid or a mixture thereof. Each of these specific buffers constitutes an alternative embodiment of the invention.

In another embodiment of the invention, the formulation further comprises a pharmaceutically acceptable preservative. In another embodiment of the invention, the formulation further comprises an isotonic agent, such as propylene glycol, mannitol or glycerol. In another embodiment of the invention, the formulation further comprises a chelating agent.

In another embodiment of the invention, the formulation further comprises a stabilizer. The use of stabilizers in pharmaceutical compositions is well known to those skilled in the art. For convenience, reference is made to Remington: The Science and Practice of Pharmacy , 19th edition, 1995.

The formation of aggregates of the multi-peptide during storage of the liquid pharmaceutical composition can adversely affect the biological activity of the multi-peptide, resulting in a loss of therapeutic efficacy of the pharmaceutical composition. In addition, aggregate formation can cause other problems, such as blocking a conduit, membrane or pump when a pharmaceutical composition containing a multi-peptide is administered using an infusion system.

The composition of the present invention is a stable liquid pharmaceutical composition, the therapeutically active component of which may exhibit aggregate formation during storage of the liquid pharmaceutical formulation It is a multi-peptide.

" Aggregate formation " means a physical interaction between molecules of a multi-peptide which results in the formation of still soluble oligomers or visible large aggregates precipitated from solution.

By "during storage" is meant that the liquid pharmaceutical composition or formulation is not administered to the individual immediately after preparation. Rather, after preparation, it is stored in a liquid form, in a frozen state, or in a dry form that is later reconstituted into a liquid form or other form suitable for administration to an individual.

The pharmaceutical composition of the present invention may further comprise an amino acid base in an amount sufficient to reduce the formation of aggregates by the multipeptide in the storage of the composition.

" Amino acid base " means an amino acid or a combination of amino acids in which any of the specified amino acids are present in the form of their free base or a salt thereof. When a combination of amino acids is used, all of the amino acids may be present in the form of their free bases, all of the amino acids may be present in the form of their salts, or some of the amino acids may be present in the form of their free bases, while other amino acids are in the form of their salts. presence. In one embodiment, the amino acid used to prepare the compositions of the present invention is an amino acid such as arginine, lysine, aspartic acid, and glutamic acid that carries a charged side chain. Any stereospecificity of a particular amino acid (eg, methionine, histidine, imidazole, arginine, lysine, isoleucine, aspartic acid, tryptophan, threonine, and mixtures thereof) The construct (i.e., L, D or a mixture thereof) or a combination of such stereoisomers may be present in the pharmaceutical composition of the present invention as long as the specific amino acid is present in its free base form or in the form of its salt. In one embodiment, the L-stereoisomer is used. The composition of the present invention can also be formulated with such derivatives of amino acids. Suitable arginine derivatives include, for example, aminoguanidine, ornithine and N-monoethyl L-arginine, suitable methionine derivatives include ethionine and butyl thioacid, and suitable cysteamine Acid derivatives include S-methyl-L cysteine. Like other amino acids, the amino acid derivative is incorporated into the composition in its free base form or as a salt thereof. In another embodiment of the invention, the amino acid or its amino acid derivative is used in a concentration sufficient to prevent or delay protein aggregation.

In another embodiment of the invention, the formulation further comprises a surfactant. In another embodiment of the invention, the formulation further comprises a protease inhibitor. The use of protease inhibitors is particularly suitable for use in pharmaceutical compositions comprising proteolytic enzymes in order to inhibit autocatalysis.

Other ingredients are likely to be present in the multi-peptide pharmaceutical formulation of the invention. Such other ingredients may include wetting agents, emulsifiers, antioxidants, accumulators, tonicity modifiers, chelating agents, metal ions, oily vehicles, proteins (eg, human serum albumin, gelatin or protein), and zwitterions (eg, Amino acids such as betaines, taurine, arginine, glycine, lysine and histidine. Of course, such other ingredients should not adversely affect the overall stability of the pharmaceutical formulations of the present invention.

The pharmaceutical composition containing the multi-peptide of the present invention can be administered to a patient in need of such treatment at several sites, for example, at a local site such as the skin and mucosa; at the bypass site, for example, intra-arterial administration, intravenous administration, and heart Intradermal administration; and in the area involving absorption, such as intradermal administration, subcutaneous administration, intramuscular administration or intraperitoneal administration.

The pharmaceutical composition of the present invention can be administered to a patient in need of such treatment via a plurality of administration routes, for example, through the tongue, sublingual, buccal, and oral. Intracavitary, oral, intragastric, nasal, transpulmonary (for example via bronchioles and alveoli or a combination thereof), epidermis, dermis, percutaneous, vaginal, rectal, transocular (eg via conjunctiva), urethra and parenteral Dosing.

The composition of the present invention can be administered in several dosage forms, for example, as a solution, suspension, emulsion, microemulsion, multiple emulsion, foam, ointment, paste, plaster, ointment, lozenge, coated lozenge, rinse, Capsules (such as hard gelatin capsules and soft gelatin capsules), suppositories, rectal capsules, drops, gels, sprays, powders, aerosols, inhalants, eye drops, ophthalmic ointments, ophthalmic rinses, vaginal uterus Support, vaginal ring, vaginal ointment, injection solution, in situ transformation solution (eg in situ gelation, in situ hardening, in situ precipitation, in situ crystallization), infusion solutions and implants.

The compositions of the present invention may be further compounded or linked, for example, via covalent, hydrophobic, and electrostatic interactions to pharmaceutical carriers, drug delivery systems, and advanced drug delivery systems to further enhance the stability of their amyloid analog derivatives, Enhance bioavailability, enhance solubility, reduce adverse effects, achieve chronotherapy well known to those skilled in the art, and enhance patient compliance or any combination thereof.

The composition of the present invention is suitable for formulating solid, semi-solid, powder and solution for pulmonary administration of derivatives of amyloid analogs, for example, using a metered dose inhaler, a dry powder inhaler and an atomizer, all devices are Those skilled in the art are familiar with the device.

The compositions of the present invention are suitable for use in controlled release, sustained release, extended release, delayed release, and slow release drug delivery systems.

By means of a syringe (as appropriate with the aid of a pen-like syringe) Parenteral administration is carried out by intramuscular, intramuscular, intraperitoneal or intravenous injection. Alternatively, parenteral administration can be carried out by means of an infusion pump. Another option is that the composition can be a solution or suspension of the amyloid analog derivative administered as a nasal or pulmonary spray. As a further option, the pharmaceutical composition comprising the multi-peptide of the present invention may also be suitable for transdermal administration (for example, by needle-free injection or patch, optionally using an iontophoretic patch), or via mucosa (eg, buccal) Within).

The multi-peptide of the present invention can be administered via the pulmonary route in the form of a solution, a suspension or a dry powder in any of the types of devices known to be suitable for pulmonary delivery of drugs. Examples of such devices include, but are not limited to, three general types of aerosols for delivery of drugs via the lung, and may include jet nebulizers or ultrasonic nebulizers, metered dose inhalers or dry powder inhalers (see Yu) J, Chien YW. Pulmonary drug delivery: Physiologic and mechanistic aspects. Crit Rev Ther Drug Carr Sys 14(4) (1997) 395-453).

In one embodiment of the invention, a pharmaceutical formulation comprising a multi-peptide of the invention is used stably for more than 6 weeks and for a stable storage period of more than 3 years.

In another embodiment of the invention, a pharmaceutical formulation comprising a multi-peptide of the invention is used stably for more than 4 weeks and for a stable storage period of more than 3 years.

In another embodiment of the invention, a pharmaceutical formulation comprising a derivative of an amyloid analog is used for a period of more than 4 weeks and is stable for more than two years.

In another embodiment of the invention, the derivative comprising an amyloid analog Biomedical formulations have been in stable use for more than 2 weeks and have been stored for more than two years.

In one aspect, a method of preparing a pharmaceutical composition comprising a derivative of the invention comprises admixing a derivative of the invention with at least one pharmaceutically acceptable excipient.

Therapeutic indications

In one aspect, the multipeptide of the present invention can be used as a pharmaceutical.

In one aspect, the multi-peptide of the present invention can be used as a medicine for treating or preventing diseases such as hyperglycemia, type 2 diabetes, abnormal glucose tolerance, type 1 diabetes, obesity, hypertension, syndrome X. , dyslipidemia, cognitive disorders, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, dyspepsia and gastric ulcer.

In one aspect, the multi-peptide is used to delay or prevent progression of type 2 diabetes.

In one aspect, the multipeptide of the present invention can be used as a medicament for treating or preventing obesity.

In one aspect, the multi-peptide of the present invention is useful as a medicament for treating or preventing hypercalcemia, osteoporosis or deformed osteitis. In one aspect, the multi-peptide is useful as a weight loss drug. In one aspect, the multi-peptide can be used as a medicine for treating or preventing obesity, or as a medicine for preventing weight gain. In one aspect, the multi-peptide can be used as a pharmaceutical product that changes the food intake (such as reducing food intake).

In one aspect, the medicament can be used to reduce food intake, reduce beta-cell apoptosis, increase beta-cell function and beta-cell population, and/or restore beta- The glucose sensitivity of the cells.

In one aspect, the multi-peptide of the invention can be used to prepare a pharmaceutical product.

In one aspect, the multi-peptide can be used to prepare a medicament for treating or preventing: hyperglycemia, type 2 diabetes, abnormal glucose tolerance, type 1 diabetes, obesity, hypertension, syndrome X , dyslipidemia, cognitive disorders, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, dyspepsia and gastric ulcer.

In one aspect, the multi-peptide can be used to prepare a medicament for delaying or preventing progression of type 2 diabetes.

In one aspect, the multi-peptide can be used to prepare glucose susceptibility for reducing food intake, reducing β-cell apoptosis, increasing β-cell function and β-cell population, and/or restoring β-cells. Pharmaceutical products.

The treatment using the multi-peptide of the present invention may also be combined with a second or more pharmacologically active substance selected, for example, from an anti-diabetic agent, an anti-obesity agent, an appetite regulating agent, an antihypertensive agent, and a therapeutic / or an agent that prevents complications caused by diabetes or is associated with diabetes, and an agent that treats and/or prevents complications and conditions caused by obesity or associated with obesity. Examples of such pharmacologically active substances are: insulin, insulin derivatives, insulin analogs, GLP-1, GLP-1 derivatives, GLP-1 analogs, acid modulating derivatives, sulfonyl urea, biguanide, Meg Meglitinide, glucosidase inhibitor, glycosidic antagonist, DPP-IV (dipeptidyl peptidase-IV) inhibitor, inhibitor of liver enzyme involved in stimulating glucose regeneration and/or hepatic glycosylation a glucose uptake regulator, a compound that modulates lipid metabolism (such as an antihyperlipidemic agent such as an HMG CoA inhibitor (statin)), reduced feeding Amount of compound, RXR agonist and agent acting on β-cell ATP-dependent potassium channel; Cholestyramine, colestipol, clofibrate, gemfibrozil Gemfibrozil), lovastatin, pravastatin, simvastatin, probucol, dextrothyroxine, neteglinide, repaglinide ; β-blockers, such as aprenolol, atenolol, timolol, pindolol, propranolol, and beauty Metoprolol; ACE (angiotensin converting enzyme) inhibitors, such as benazepril, captopril, enalapril, fosinopril , lisinopril, alatriopril, quinapril and ramipril; calcium channel blockers such as nifedipine, felodipine (felodipine), nicardipine, isradipine, nimodipine, diltiazem And verapamil; and alpha-blockers such as doxazosin, urapidil, prazosin and terazosin; CART ( Cocaine amphetamine-regulated transcript) agonist, NPY (neuropeptide Y) antagonist, MC4 (melanocortin 4) agonist, appetite hormone antagonist, TNF (tumor necrosis factor) agonist, CRF (corticotropin release) Factor) agonist, CRF BP (corticotropin releasing factor binding protein) antagonist, urotensin agonist, β3 agonist, MSH (melanocyte stimulating hormone) agonist, MCH (melanocyte Antagonistic hormone Agent, CCK (cholinergic) agonist, serotonin reuptake inhibitor, serotonin and norepinephrine reuptake inhibitor, mixed serotonin and norepinephrine-exciting compound, 5HT (serotonin) An agonist, bombesin agonist, galanin antagonist, growth hormone, growth hormone releasing compound, TRH (thyrotropin releasing hormone) agonist, UCP 2 or 3 (uncoupling protein 2 or 3) Regulators, leptin agonists, DA agonists (bromocriptin, doprexin), lipase/amylase inhibitors, RXR (retinoid X receptor) modulators , TR β agonist; histamine H3 antagonist, gastrin and gastrin analog.

It will be understood that any suitable combination of the multi-peptide of the present invention with one or more of the above compounds, and optionally one or more other pharmacologically active substances, is considered to be within the scope of the invention.

The invention is further summarized in the following paragraphs:

A multi-peptide comprising: (i) a multi-peptide comprising an amino acid sequence of the analog of SEQ ID No: 2, wherein: the analog comprises a glutamic acid residue at position 14; The amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2; and the restriction is that the multi-peptide is not N-α-(19-carboxyundecyl)-Glu-Glu -Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; and the multi-peptide is optionally attached to at least one of its at least one amino acid residue.

(ii) multiple wins comprising the amino acid sequence of the SEQ ID No: 2 analog a peptide, wherein: (a) the analog comprises a glutamic acid residue at position 14; wherein the amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2; and the restriction is The multi-peptide is not N-α-(19-carboxynonanoyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; and (b) the multi-peptide is at pH 4 having a solubility of about 100 μM or greater than 100 μM, the multipeptide is optionally attached to at least one of its at least one amino acid residue.

(iii) a multi-peptide comprising the amino acid sequence of the analog of SEQ ID No: 2, wherein: (a) the analog comprises a glutamic acid residue at position 14; wherein the amino acid sequence number of the analog Corresponding to the amino acid numbering sequence of SEQ ID No: 2; and the restriction is that the multi-peptide is not N-α-(19-carboxynonanoyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; and (b) the multi-peptide has a solubility of about 100 μM or greater at pH 7, and the multi-peptide is optionally attached to at least one of its amino acid residues A substituent.

(iv) a multi-peptide comprising the amino acid sequence of the analog of SEQ ID No: 2, wherein: (a) the analog comprises a glutamic acid residue at position 14; Wherein the amino acid sequence number of the analog corresponds to the amino acid numbering sequence of SEQ ID No: 2; and the restriction is that the multi-peptide is not N-α-(19-carboxynodecyl)-Glu -Glu-Arg-[Glu1, Glu14, His17, Glu31]- pramlintide; and (b) the multi-peptide has a solubility of about 100 μM or more at pH 4; and (c) the multi-peptide Having a solubility of about 100 μM or greater than 100 μM at pH 7 The multi-peptide is optionally attached to at least one of its at least one amino acid residue.

(v) a multi-peptide comprising the amino acid sequence of the analog of SEQ ID No: 2, wherein: (a) the analog comprises a glutamic acid residue at position 14; wherein the amino acid sequence number of the analog Corresponding to the amino acid numbering sequence of SEQ ID No: 2; and the restriction is that the multi-peptide is not N-α-(19-carboxynonanoyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]-pramlintide; and (b) the multi-peptide has a solubility of about 100 μM or greater at pH 4; and (c) the multi-peptide has about 100 μM or greater at pH 7. a solubility of 100 μM; and (d) the multi-peptide has a physical stability of about 25 hours or more than 25 hours in the fibril formation assay; the multi-peptide is optionally attached to at least one of its amino acid residues A substituent.

2. The multipeptide of paragraph 1, wherein the analog comprises a residue at position 17, the residue being independently selected from any one of the group consisting of histidine, arginine, lysine, and proline; Preferred are histamine and arginine; more preferably arginine.

3. The multi-peptide of any of paragraphs 1 and 2, wherein the analog comprises a residue at position 37, the residue being independently selected from any of valine and tyrosine; preferably It is a proline.

4. The multi-peptide of any of the preceding paragraphs, wherein the analog comprises a residue at position 35, the residue being independently selected from the group consisting of histidine, arginine, lysine, aspartic acid, glutamic acid Any one of aspartic acid and glutamic acid; preferably histidine, aspartic acid and glutamic acid; more preferably histidine and aspartic acid; more preferably Aspartic acid.

5. The multi-peptide of any of the preceding paragraphs, wherein the analog comprises a residue at position 1, which residue is deleted or independently selected from the group consisting of alanine, cysteine, glutamic acid, glycine, histamine Any of acid, arginine, serine and lysine; preferably glycine, histidine, arginine and lysine; more preferably lysine.

6. The multi-peptide of any of the preceding paragraphs, wherein the analog comprises a residue at position 3, the residue being independently selected from the group consisting of glycine, histidine, arginine, serine and aspartame Any of the acids; preferably aspartic acid.

7. The multi-peptide of any of the preceding paragraphs, wherein the analog comprises a residue at position 18, the residue being independently selected from any one of arginine, lysine, and histidine; preferably Histidine.

8. The multi-peptide of any of the preceding paragraphs, wherein the analog comprises a position a residue at 21 independently selected from any one of alanine, lysine, glutamic acid, serine, threonine, and aspartic acid; preferably glutamine Acid, aspartic acid; more preferably aspartic acid.

9. The multi-peptide of any of the preceding paragraphs, wherein the analog comprises a residue at position 22, the residue being independently selected from the group consisting of glutamic acid, glutamic acid, serine, threonine and aspartame Any of the amine acids; preferably seric acid and aspartic acid; more preferably aspartic acid.

10. The multi-peptide of any of the preceding paragraphs, wherein the analog comprises a residue at position 26, the residue being independently selected from any of lysine, arginine and isoleucine; preferably For aspartic acid.

11. The multi-peptide of any of the preceding paragraphs, wherein the analog comprises a residue at position 31, the residue being independently selected from the group consisting of serine, glutamic acid, aspartic acid and aspartic acid Any one; preferably aspartic acid and aspartic acid; more preferably aspartic acid.

12. The multi-peptide of any of the preceding claims, wherein the analog comprises an amino acid residue at position 14 for glutamic acid, a residue at position 17 for arginine, and a position at 37 It is the residue of proline.

13. The peptide according to paragraphs 1 to 11, wherein the analog comprises an amino acid residue of glutamic acid at position 14, a residue of histidine at position 17, and a proline at position 37. The residue.

14. The peptide according to paragraphs 1 to 11, wherein the analog comprises an amino acid residue at position 14 as glutamic acid, a residue at position 17 of arginine, and a histidine at position 35. The residue.

15. The peptide according to paragraphs 1 to 11, wherein the analog comprises a position 14 is the amino acid residue of glutamic acid, the residue of histidine at position 17 and the residue of histidine at position 35.

16. The multi-peptide of any of the preceding claims, wherein the remaining residues are the same as in SEQ ID No: 2.

17. The multi-peptide of any of the preceding claims, wherein the analog comprises an amine group at positions 2, 4 to 16, 19, 20, 23 to 25, 27 to 30, 32 to 34 and 36. The acid residue is the same as the residue at positions 2, 4 to 16, 19, 20, 23 to 25, 27 to 30, 32 to 34 and 36 of SEQ ID No: 2.

18. The multipeptide of paragraph 1, which comprises an amino acid sequence having an analog of SEQ ID No: 2 of formula (I): Xaa ₁ -Cys-Xaa ₃ -Thr-Ala-Thr- Cys-Ala-Thr-Gln-Arg-Leu-Ala-Glu-Phe-Leu-Xaa ₁₇ -Xaa ₁₈ -Ser-Ser-Xaa ₂₁ -Xaa ₂₂ -Phe-Gly-Pro-Xaa ₂₆ -Leu-Pro-Pro -Thr-Xaa ₃₁ -Val-Gly-Ser-Xaa ₃₅ -Thr-Xaa ₃₇ ; Formula (I) (SEQ ID No: 3) wherein Xaa _{1 is} deleted or independently selected from Ala, Cys, Glu, Gly, His, Arg, Ser and Lys; Xaa _{3 is} independently selected from Gly, His, Arg, Ser and Asn; Xaa _{17 is} independently selected from the group consisting of His, Arg, Lys and Val; Xaa _{18 is} independently selected from Arg, Lys and His; Xaa ₂₁ Independently selected from the group consisting of Ala, Lys, Gln, Ser and Asn; Xaa _{22 is} independently selected from the group consisting of Glu, Gln, Ser, Thr and Asn; Xaa _{26 is} independently selected from the group consisting of Pro, Arg and Ile; Xaa _{31 is} independently selected from the group consisting of Ser, Glu, Asp and Asn; Xaa _{35 are} independently selected from the group consisting of His, Arg, Lys, Asp and Glu; Xaa _{37 is} independently selected from Pro and Tyr; and wherein the C-terminus can be derivatized as appropriate;

19. The multipeptide of paragraph 18, wherein Xaa ₁₇ is Arg or His.

20. The multi-peptide of any of paragraphs 18 and 19, wherein Xaa ₃₇ is Pro.

21. The multi-peptide of any of paragraphs 18 to 20, wherein Xaa ₃₅ is His.

22. The multi-peptide of any one of paragraphs 18 to 20, wherein: Xaa ₁ is Lys; Xaa ₃ is Asn; Xaa ₁₇ is His or Arg; Xaa ₁₈ is His; Xaa ₂₁ is Asn; Xaa ₂₂ is Asn; Xaa ₂₆ is Ile; Xaa ₃₁ is Asn; Xaa ₃₅ is His or Asn; and Xaa ₃₇ is Pro.

23. The peptide according to paragraph 18, wherein Xaa _{1 is} deleted or independently selected from the group consisting of His, Arg and Lys; Xaa _{3 is} independently selected from the group consisting of Gly, His and Asn; Xaa _{17 is} independently selected from the group consisting of His, Arg and Val; Xaa ₁₈ independently selected from Arg and His; Xaa _{21 is} independently selected from Ser and Asn; Xaa ₂₂ is Asn; Xaa ₂₆ is Ile; Xaa _{31 is} independently selected from Glu and Asn; Xaa _{35 is} independently selected from His, Arg, Lys , Asp and Glu; Xaa _{37 are} independently selected from Pro and Tyr.

24. The multipeptide of paragraph 18, wherein Xaa _{1 is} deleted or independently selected from the group consisting of Gly, His, Arg, Ser, and Lys; Xaa _{3 is} independently selected from the group consisting of His and Asn; and Xaa _{17 is} independently selected from the group consisting of His, Arg, and Val. Xaa _{18 is} independently selected from Arg and His; Xaa _{21 is} independently selected from Gln, Ser and Asn; Xaa ₂₂ is Asn; Xaa _{26 is} independently selected from Pro and Ile; Xaa ₃₁ is Asn; Xaa ₃₅ is His; Xaa ₃₇ For Tyr.

25. The peptide according to paragraph 18, wherein Xaa _{1 is} deleted or independently selected from the group consisting of Ala, Cys, Glu, Gly, His, Arg, Ser and Lys; Xaa ₃₁ is Asn; Xaa _{17 is} independently selected from the group consisting of His, Arg and Val; Xaa _{18 is} independently selected from Arg, Lys and His; Xaa _{21 is} independently selected from Gln and Asn; Xaa _{22 is} independently selected from Thr and Asn; Xaa ₂₆ is Ile; Xaa ₃₁ is Asn; Xaa _{35 is} independently selected from Gly and Asn; Xaa ₃₇ and Pro.

26. The multipeptide of paragraph 18, wherein Xaa _{1 is} deleted; Xaa _{3 is} independently selected from the group consisting of Gly and Asn; Xaa _{17 is} independently selected from the group consisting of His, Arg and Val; Xaa _{18 is} independently selected from the group consisting of Arg and His; Xaa _{21 is} independent Is selected from Gln and Asn; Xaa _{22 is} independently selected from Gln and Asn; Xaa ₂₆ is Ile; Xaa _{31 is} independently selected from Glu and Asn; Xaa _{35 is} independently selected from His and Ser; Xaa _{37 is} independently selected from Pro and Tyr.

27. The peptide according to paragraph 18, wherein Xaa ₁ is Lys; Xaa ₃ is Asn; Xaa _{17 is} independently selected from the group consisting of His, Lys, Arg and Val; Xaa _{18 is} independently selected from Arg and His; Xaa _{21 is} independently selected From Ala, Lys, Gln and Ser; Xaa _{22 is} independently selected from Glu, Gln, Ser, Thr and Asn; Xaa _{26 is} independently selected from Pro and Ile; Xaa _{31 is} independently selected from Ser, Glu, Asp and Asn; Xaa _{35 is} independently selected from the group consisting of His, Glu and Asn; Xaa _{37 is} independently selected from the group consisting of Pro and Tyr.

28. The multi-peptide of any of the preceding paragraphs which comprises a C-terminal decylamine.

29. The multipeptide of paragraph 28, wherein the C-terminal guanamine has the formula (II): (II) C(O)NR ¹ R ² wherein R ¹ and R ^{2 are} independently selected from H and alkyl. R ¹ and R ^{2 are} preferably both H.

30. A peptide as much of paragraph 29, wherein R ¹ and R ² are both H.

31. The multi-peptide of any of the preceding paragraphs, wherein at least one substituent is attached to one of the amino acid residues.

32. The multipeptide of paragraph 31, wherein the substituent is selected from the group consisting of a hydrocarbyl group, a hydroxyl group, and a halogen atom.

33. The multi-peptide of any of paragraphs 31 and 32, wherein the substituent has the formula (II): (I1) L _n -Y wherein L is a linker; n = 0 or 1 Y is an albumin binding moiety .

34. The multipeptide of paragraph 33, wherein the albumin binding moiety is a thiol group selected from the group consisting of: (a) CH ₃ (CH ₂ ) _r CO-, wherein r is an integer from 12 to 20; (b) HOOC (CH ₂ ) _s CO-, wherein s is an integer from 12 to 22, or s is an integer from 12 to 18, or s is from 16 to 18, or s is preferably 18.

The multi-peptide of any one of paragraphs 33 and 34, wherein the linker is selected from the group consisting of γGlu, γGlu-γGlu, γGlu-γGlu-γGlu, γGlu-γGlu-γGlu-γGlu, Glu, Glu-Glu, Glu-γGlu, Glu-Arg, Glu-Glu-Arg, His, His-His, His-γGlu, His-His-γGlu, Gly, Gly-γGlu, Ser, Ser-γGlu, D-Arg- D-Arg, Arg, Arg-Arg, Arg-Arg-γGlu, Ser-Ser, -Gly-Ser-Ser, Ser-Ser, -Gly-Ser-Ser-γGlu, Ser-Ser-Gly-Ser-Ser- Gly and Ser-Ser-Gly-Ser-Ser-Gly-γGlu, γGlu-OEG, γGlu-OEG-OEG and OEG, the linker is preferably selected from the group consisting of γGlu, γGlu-γGlu, γGlu-OEG, γGlu-OEG-OEG And OEG, the linker is more preferably γGlu.

36. The multi-peptide of any of the preceding paragraphs, wherein the substituent is selected from the group provided in Table 1 (provided above).

37. The multi-peptide of any of the preceding paragraphs, wherein the substituent is attached to the alpha-amino group of the N-terminal amino acid residue or to the Lys residue.

38. The multi-peptide of any of the preceding paragraphs wherein the substituent is attached only to the amino acid residue at position 1.

39. A multi-peptide according to any of the preceding paragraphs selected from the group consisting of any of the multi-peptides provided in Table 2 (provided above); or selected from any of the multi-peptides provided in Table 3 (provided above) a group consisting of; or selected from the group consisting of any of the peptides provided in Table 4 (provided above); or selected from the group consisting of any of the peptides provided in Table 5 (provided above); or selected from the list a group of any of the multi-peptides provided in 6 (provided previously); or selected from a group consisting of any of the multi-peptides provided in Table 7 (provided previously); or a group consisting of any of the multi-peptides provided in Table 8 (provided previously); or selected from Table 9 (provided above) Any of a plurality of peptide compositions provided; or selected from the group consisting of any of the peptides provided in Table 10 (provided above); or selected from any of the peptides provided in Table 11 (provided above) a group; or a group consisting of any of the multi-peptides provided in Table 12 (provided previously); or a group consisting of any of the multi-peptides provided in Table 13 (provided above); or selected from Table 14 Any of the multi-peptide compositions provided (provided above); or selected from the group consisting of any of the multi-peptides provided in Table 15 (provided previously).

40. A multi-peptide according to any of the preceding paragraphs which is selected from the group consisting of any of the multi-peptides provided in Table 8 (provided above).

41. A multi-peptide according to any of the preceding paragraphs which is selected from the group consisting of any of the multi-peptides provided in Table 9 (provided above).

42. The multi-peptide of any of the preceding paragraphs, wherein the multi-peptide is: N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, Arg17 , Pro37]-pramlintide [Example 53].

43. A pharmaceutical composition comprising a multi-peptide of any of the preceding paragraphs and a pharmaceutically acceptable excipient.

44. The pharmaceutical composition of paragraph 43, which is suitable for parenteral administration.

45. A method of preparing a pharmaceutical composition according to paragraph 43 or paragraph 44, which comprises mixing a multi-peptide of any of the preceding paragraphs with at least one pharmaceutically acceptable excipient.

46. A multi-peptide according to any of the preceding paragraphs, which is suitable for use as a pharmaceutical.

47. A multi-peptide according to any of the preceding paragraphs, which is useful for the treatment or prevention of hyperglycemia, type 2 diabetes, abnormal glucose tolerance, type 1 diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive disorders, Atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, dyspepsia and gastric ulcers.

48. A multi-peptide according to any of the preceding paragraphs, which is suitable for delaying or preventing progression of a disease of type 2 diabetes.

49. A multi-peptide according to any of the preceding paragraphs, which is suitable for use in the prevention or treatment of obesity.

50. A multi-peptide according to any of the preceding paragraphs, which is suitable for reducing food intake, reducing beta cell apoptosis, increasing beta-cell function and beta-cell population, and/or restoring glucose sensitivity of beta cells.

51. The multi-peptide of any of the preceding paragraphs, which is useful for treating or preventing hypercalcemia, osteoporosis or malformation osteitis.

52. A method for treating or preventing a disease by administering a multi-peptide of any of the preceding paragraphs to an animal: hyperglycemia, type 2 diabetes, abnormal glucose tolerance, type 1 diabetes, obesity, hypertension, Syndrome X, dyslipidemia, cognitive disorders, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, dyspepsia and gastric ulcer.

53. A method of delaying or preventing progression of a type 2 diabetes disease by administering to a animal a multi-peptide as in any of the preceding paragraphs.

54. A method of reducing food intake, reducing β-cell apoptosis, increasing β-cell function and β-cell population, and/or restoring glucose sensitivity of β-cells The method of administering a multi-peptide of any of the preceding paragraphs to an animal.

55. A method of treating or preventing hypercalcemia, osteoporosis or deformed osteitis by administering to a animal a multi-peptide as in any of the preceding paragraphs.

56. any preceding Paragraph many peptide, wherein the plurality of human amylin peptide receptor of about 1800 pM, or less than 1800 pM of EC _50.

57. any preceding Paragraph many peptide, wherein the peptide has a plurality of approximately 1800 pM or human calcitonin receptor of less than 1800 pM EC _50.

The invention is further summarized in the following other paragraphs:

An amyloid derivative which is an amyloid analog having up to ten amino acid residue modifications as compared to SEQ ID NO: 2 and which is an alpha-terminal amino acid residue of the amyloid analog a substituent attached to the amine or Lys residue, wherein the substituent comprises an albumin binding moiety, and wherein a. the amino acid residue at position 14 of the amyloid analog is Glu or b. The amyloid is similar The amino acid residue at position 35 is His, Arg, Lys, Asp or Glu or c. The amino acid residue at position 37 of the amyloid analog is Pro.

2. The derivative of paragraph 1, wherein the amino acid residue at position 14 is Glu.

3. A derivative according to paragraphs 1 to 2, wherein a. the amino acid residue at position 14 is Glu and the amino acid at position 35 is His, Arg, Lys, Asp or Glu or b. Position 14 The amino acid residue is Glu and the amino acid at position 37 is Pro.

4. The derivatives of paragraphs 1 to 3, wherein a. The amino acid residue at position 14 is Glu and the amino acid at position 35 is His.

5. The derivative of paragraphs 1 to 4, wherein the amino acid residue at position 17 is His.

6. The derivative of paragraphs 1 to 5, wherein the amyloid analog comprises 1, 2, 3, 4, 5 or 6 substitutions.

7. The derivative of paragraphs 1 to 6, wherein the amino acid at position 1 of the amyloid analog is substituted or deleted.

8. The derivative of paragraphs 1 to 7, wherein the amino acid at position 1 is selected from the group consisting of Lys, Glu, Arg, Ala, Ser, Cys, Gly and His.

9. The derivative of paragraphs 1 to 8, wherein the substituent comprises a linker.

10. The derivative of paragraphs 1 to 9, wherein the linker comprises from 1 to 10 amino acids attached to the a-amino or Lys residue of the N-terminal amino acid residue of the amyloid analog.

11. The derivative of paragraph 10, wherein the amino acid is selected from the group consisting of γGlu, γGlu-γGlu, γGlu-γGlu-γGlu, γGlu-γGlu-γGlu-γGlu, Glu, Glu-Glu, Glu -γGlu, Glu-Arg, Glu-Glu-Arg, His, His-His, His-γGlu, His-His-γGlu, Gly, Gly-γGlu, Ser, Ser-γGlu, D-Arg-D-Arg, Arg , Arg-Arg, Arg-Arg-γGlu, Ser-Ser, -Gly-Ser-Ser, Ser-Ser, -Gly-Ser-Ser-γGlu, Ser-Ser-Gly-Ser-Ser-Gly and Ser-Ser -Gly-Ser-Ser-Gly-γGlu.

12. The derivative of paragraphs 1 to 11, wherein the linker comprises -C(O)-(CH ₂ ) _l -O-[CH ₂ CH ₂ -O] _m -(CH ₂ ) _p -[NHC(O )-(CH ₂ ) _l -O-[(CH ₂ ) _n -O] _m -(CH ₂ ) _p ] _q -NH-, wherein l, m, n and p are independently from 1 to 7, and q is 0 to 5.

13. The derivative of paragraphs 1 to 12, wherein the linker is selected from the group consisting of: -C(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -NH- Or -C(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -[NHC(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -] ₁ NH- and -C(O)-(CH ₂ ) ₂ -O-[CH ₂ CH ₂ -O] ₇ -(CH ₂ ) ₂ -NH-.

14. The derivative of paragraphs 1 to 13, wherein the linker is selected from the group consisting of γGlu-C(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -[ NHC(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -] ₁ -NH- and Arg-Arg-γGlu-C(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -[NHC(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -] ₁ -NH-.

The derivative according to any one of paragraphs 1 to 14, wherein the albumin-binding residue is a thiol group selected from the group consisting of HOOC(CH ₂ ) _s CO-, wherein s is an integer from 14 to 20, for example 16 or 18.

16. The derivative of paragraphs 1 to 15, wherein the derivative comprises a γGlu linker linked to the N-terminus of the amyloid analog and HOOC(CH ₂ ) ₁₈ CO or HOOC (CH ₂ ) as the albumin-binding residue ₁₆ CO-, and wherein the sequence of the amyloid analog comprises: a. Glu at position 14 or His or Arg c at position 17. His at position 35 or Pro at position 37.

17. The derivative of paragraph 1, wherein the amyloid analog comprises the amino acid sequence of formula 1: Xaa ₁ -Cys-Xaa ₃ -Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu- Ala-Glu-Phe-Leu-Xaa ₁₇ -Xaa ₁₈ -Ser-Ser-Xaa ₂₁ -Xaa ₂₂ -Phe-Gly-Pro-Xaa ₂₆ -Leu-Pro-Pro-Thr-Xaa ₃₁ -Val-Gly-Ser- Xaa ₃₅ -Thr-Xaa ₃₇

Formula (I) (SEQ ID No: 3)

Wherein Xaa _{1 is} deleted or independently selected from the group consisting of Ala, Cys, Glu, Gly, His, Arg, Ser and Lys; Xaa _{3 is} independently selected from Gly, His, Arg, Ser and Asn; Xaa _{17 is} independently selected from His, Arg , Lys and Val; Xaa _{18 is} independently selected from Arg, Lys and His; Xaa _{21 is} independently selected from the group consisting of Ala, Lys, Gln, Ser and Asn; Xaa _{22 is} independently selected from Glu, Gln, Ser, Thr and Asn; Xaa _{26 is} independently selected from the group consisting of Pro, Arg and Ile; Xaa _{31 is} independently selected from the group consisting of Ser, Glu, Asp and Asn; Xaa _{35 is} independently selected from the group consisting of His, Arg, Lys, Asp and Glu; Xaa _{37 is} independently selected from Pro and Tyr And wherein the C-terminus can be derived as a guanamine.

18. The derivative according to paragraphs 1 and 17, wherein Xaa _{1 is} deleted or independently selected from the group consisting of His, Arg and Lys; Xaa _{3 is} independently selected from the group consisting of Gly, His and Asn; Xaa _{17 is} independently selected from the group consisting of His, Arg and Val; Xaa _{18 is} independently selected from Arg and His; Xaa _{21 is} independently selected from Ser and Asn; Xaa ₂₂ is Asn; Xaa ₂₆ is Ile; Xaa _{31 is} independently selected from Glu and Asn; Xaa _{35 is} independently selected from His, Arg, Lys, Asp, and Glu; Xaa _{37 are} independently selected from Pro and Tyr.

19. The derivative of any of paragraphs 1 and 17, wherein Xaa _{1 is} deleted or independently selected from the group consisting of Gly, His, Arg, Ser, and Lys; Xaa _{3 is} independently selected from the group consisting of His and Asn; and Xaa _{17 is} independently selected from the group consisting of His, Arg and Val; Xaa _{18 are} independently selected from Arg and His; Xaa _{21 is} independently selected from Gln, Ser and Asn; Xaa ₂₂ is Asn; Xaa _{26 is} independently selected from Pro and Ile; Xaa ₃₁ is Asn; Xaa ₃₅ It is His; Xaa ₃₇ is Tyr.

20. The derivative according to paragraphs 1 and 17, wherein Xaa _{1 is} deleted or independently selected from the group consisting of Ala, Cys, Glu, Gly, His, Arg, Ser and Lys; Xaa ₃ is Asn; Xaa _{17 is} independently selected from His, Arg And Val; Xaa _{18 is} independently selected from Arg, Lys and His; Xaa _{21 is} independently selected from Gln and Asn; Xaa _{22 is} independently selected from Thr and Asn; Xaa ₂₆ is Ile; Xaa ₃₁ is Asn; Xaa _{35 is} independently selected From Gly and Asn; Xaa ₃₇ is Pro.

The derivative according to any one of paragraphs 1 and 17, wherein Xaa _{1 is} deleted; Xaa _{3 is} independently selected from Gly and Asn; Xaa _{17 is} independently selected from the group consisting of His, Arg and Val; Xaa _{18 is} independently selected from Arg and His; Xaa _{21 is} independently selected from Gln and Asn; Xaa _{22 is} independently selected from Gln and Asn; Xaa ₂₆ is Ile; Xaa _{31 is} independently selected from Glu and Asn; Xaa _{35 is} independently selected from His and Ser; Xaa _{37 is} independent It is selected from Pro and Tyr.

22. The derivative according to paragraphs 1 and 17, wherein Xaa ₁ is Lys; Xaa ₃ is Asn; Xaa _{17 is} independently selected from the group consisting of His, Lys, Arg and Val; Xaa _{18 is} independently selected from Arg and His; Xaa ₂₁ independently Is selected from the group consisting of Ala, Lys, Gln and Ser; Xaa _{22 is} independently selected from the group consisting of Glu, Gln, Ser, Thr and Asn; Xaa _{26 is} independently selected from Pro and Ile; Xaa _{31 is} independently selected from the group consisting of Ser, Glu, Asp and Asn; Xaa _{35 is} independently selected from the group consisting of His, Glu and Asn; Xaa _{37 is} independently selected from Pro and Tyr.

23. The derivative of paragraphs 1 and 17 to 22, wherein the albumin binding residue is non-covalently bound to albumin.

The derivative of any of paragraphs 1 and 17 to 23, wherein the albumin binding residue has a binding affinity for human serum albumin of less than about 10 μM or less than about 1 μM.

25. The derivative of any of paragraphs 1 and 17 to 24, wherein the albumin binding residue comprises a group that is negatively chargeable at pH 7.4.

26. The derivative of any of paragraphs 1 and 17 to 25, wherein the albumin binding residue comprises a carboxylic acid group.

27. The derivative of any of paragraphs 1 and 17 to 26, wherein the albumin binding residue is a thiol group selected from the group consisting of HOOC(CH ₂ ) _s CO-, wherein s is an integer from 12 to 22. For example, 17, 18, 19, 20, 21 or 22.

28. A derivative according to paragraphs 1 and 27, wherein s is 16 or 18.

29. The derivative of paragraphs 1 and 17 to 28, wherein the substituent comprises a linker.

30. The derivative of paragraphs 1 and 17 to 29, wherein the linker comprises from 1 to 10 amino acids attached to the a-amino or Lys residue of the N-terminal amino acid residue of the amyloid analog .

31. The derivative according to paragraphs 1 and 17 to 30, wherein the amino acid is selected from the group consisting of γGlu, γGlu-γGlu, γGlu-γGlu-γGlu, γGlu-γGlu-γGlu-γGlu, Glu, Glu -Glu, Glu-γGlu, Glu-Arg, Glu-Glu-Arg, His, His-His, His-γGlu, His-His-γGlu, Gly, Gly-γGlu, Ser, Ser-γGlu, D-Arg-D -Arg, Arg, Arg-Arg, Arg-Arg-γGlu, Ser-Ser, -Gly-Ser-Ser, Ser-Ser, -Gly-Ser-Ser- γGlu, Ser-Ser-Gly-Ser-Ser-Gly and Ser-Ser-Gly-Ser-Ser-Gly-γGlu.

32. The derivative of paragraphs 1 and 17 to 31, wherein the linker comprises γGlu.

33. The derivative of paragraphs 1 and 17 to 32, wherein the linker comprises -C(O)-(CH ₂ ) _l -O-[CH ₂ CH ₂ -O] _m -(CH ₂ ) _p -[NHC (O)-(CH ₂ ) _l -O-[(CH ₂ ) _n -O] _m -(CH ₂ ) _p ] _q -NH-, wherein l, m, n and p are independently from 1 to 7, and q is 0 to 5.

34. The derivative of paragraphs 1 and 17 to 33, wherein the linker is selected from the group consisting of: -C(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ - NH- or -C(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -[NHC(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ - CH ₂ -] ₁ NH- and -C(O)-(CH ₂ ) ₂ -O-[CH ₂ CH ₂ -O] ₇ -(CH ₂ ) ₂ -NH-.

35. The derivative of paragraphs 1 and 17 to 34, wherein the linker is selected from the group consisting of γGlu-C(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -[NHC(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -] ₁ -NH- and Arg-Arg-γGlu-C(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -[NHC(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -] ₁ -NH-.

36. The derivative of paragraphs 1 and 17 to 28, wherein the linker is γGlu-γGlu-γGlu-γGlu-C(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ - [NHC(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -] ₁ -NH-.

37. The derivative of paragraph 1, wherein the derivative is selected from the group consisting of N-α-[(S)-4-carboxy-4-(19-carboxyundecylamino)butane -[His1,Glu14,His17,Arg18]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[His1,Glu8, Glu14,His17,Arg18]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[des1, Gly3, Glu14, His17, Arg18, Glu31, Gly35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[des1, Gly3, Glu14, His17, Arg18, Gln21, Gln22, Glu31, Gly35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu1, Glu14, His17, Arg18]-Plan Linpeptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[His1, Glu14, His17, Arg18, Ala25, Pro26, Ser28, Ser29]-Plan Linpeptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[His1, Glu14, His17, Arg18, Glu25, Ser28, Arg29]-planin , N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1, Glu14, His17, Arg18, Glu22, Ala25, Arg26, Ser28, Ser29]-Plan Linpeptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[His1, Glu14, His17, Arg18, Ser21, Ser22, Asp31, Asp35]-Plan Lin peptide, N-α-[(S)-4-carboxy-4-(19 -carboxy-nonadesylamino)butanyl]-[Glu1, Glu14, His17, Arg18, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-His-His-[His1,Glu14,His17]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-His-His-[His1,Glu14,Arg17]-planin, N-ε-21-[2-(2- {2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptylamino)butaninyl]ethoxy}ethoxy)acetamido Ethoxy}ethoxy)ethinyl]-[Arg1, Glu14, His17, Arg18, Lys21]-planin, N-α-[(S)-4-carboxy-4-(19-carboxyl Nonadenyl)butanyl]-[His1,Glu14,Arg18]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[His1 , Gly3, Glu14, His17, Arg18, Ser21, Ser22, Glu31, Glu35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]- His-[His1, Glu14, His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[His1,Glu14,His17]- Pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-His-His-[His1, Glu14, His17, Ala25, Pro26, Ser28, Ser29 ]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-His-His-[His1, Glu14] - pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-His-His-[His1, Glu14, His17, Arg35]-planin , N-α-[(S)-4-carboxy-4-(19-carboxyundecylamino)butane ]]-His-His-[His1, Arg3, Glu14, His17]- pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-) Carboxy-4-(19-carboxyheptadecylamido)butanylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[His1,Glu14, His17, Arg18, Ala19, Thr20, Gln21, Glu22, Leu23]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu1, Gly3, Glu14, Arg18, Ser21, Ser22, Ala25, Pro26, Ser28, Ser29, Glu31, Arg35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonadecanamide Butyl]-Glu-[Arg1, Glu14, His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-[ Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14,His17]-pramlintide, N-α -[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[des1,Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy- 4-(19-carboxynonylamido)butanyl]-[Glu1, Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonadecanamide Butyl]-[Glu1, Glu14, His17, Arg18]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxy-ten Amidino)butanyl]-His-His-[His1,Glu14,His17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino) Dingshen]-His-His-[His1, His3, Glu14, His17, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxyundecylamino)butanyl]-His-His-[His1, His3, Glu14, His17, His29, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxyundecylamino)butanyl]-His-His-[His1, Glu14, His17, His29, His35]-pramlintide, N- --[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-His-His-[His1,His3,Glu14,His17]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-His-His-[His1,Glu14,His17,His29]-planin, N-α-[2-(2- {2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptylamino)butaninyl]ethoxy}ethoxy)acetamido Ethoxy}ethoxy)ethinyl]-D-Arg-D-Arg-[Glu14,His17,Arg18]-planin, N-α-[2-(2-{2-[2- (2-{2-[(S)-4-carboxy-4-(19-carboxyheptadecanosamino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}B Oxy)ethinyl]-D-Arg-D-Arg-[Glu14,His17,Arg18,Ala21,Ser35]-pramlintide, N-α-[(S)-4-carboxy-4-(19- Carboxylamidylamino)butanyl]-Ser-Ser-Gly-Ser-Ser-[Gly1,Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19- Carboxyylammonium)butyryl]- Glu-[His1, Glu14, His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-Glu-[Glu14,His17 ]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-Glu-[Glu14,His17]-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadecanosyl)butaninyl] ethoxylate Ethyloxyacetate]ethoxy}ethoxy)ethinyl]-[des1,Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4 -(19-carboxynonylamino)butanyl]-Ser-Ser-Gly-Ser-Ser-Gly-[His1,Glu14,His17]-pramlintide, N-α-[(S)-4-carboxyl -4-(17-carboxyheptadelamido)butanyl]-His-His-[His1,Glu14,Hi17]-pramlintide, N-α-[(S)-4-carboxy-4-(17- Carboxythylideneamino)butanyl]-[Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptylamino)butanyl]-His- [His1, Glu14, His17]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-[His1,Glu14,His17]-Planin Peptide, N-α-{(S)-4-carboxy-4-[(S)-4-carboxy-4-(17-carboxyheptylamino)butaninyl]butanyl}-[des1, Glu14 ,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14,Arg17]-pramlintide, N-α- [(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-[Glu14,Arg17]-pramlintide, N-α-[(S)-4-carboxy-4-( 19-carboxy nineteen Amino)butanyl]-[Glu14,His17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, Arg17 ,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxyundecylamino)butane Base]-[Glu10, Glu14, His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14,His17,His35] -Pullinin, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Glu-[Glu14,His17,His35]-pramlintide, N-α -[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, Arg17, His35]-pramlintide, N-α-[(S)-4-carboxy- 4-(19-carboxynonylamido)butanyl]-Glu-[Glu14,Arg17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxy seventeen醯Amino)butanyl]-[Glu14,His17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptylamino)butanyl]-Glu-[ Glu14, His17, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-[Glu14, Arg17, His35]-pramlintide , N-α-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-Glu-[Glu14, Arg17, His35]-planin, N-α-[(S )-4-carboxy-4-(19-carboxynonylamido)butanyl]-His-His-[His1, Glu14, His17, Ser21, Glu35]-pramlintide, N-α-[(S)- 4-carboxy-4-(19-carboxyundecylamino)butanyl]-[Glu14, His17, Ala25, Pro26, Ser28 ,Ser29,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, Arg17, Ala25, Pro26, Ser28, Ser29 ,His35]-Planlin Peptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-His-[Glu14,His17,His35]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-His-His-[Glu14,His17,His35]-pramlintide, N-α-[(S)-4-carboxyl 4-(19-carboxynonylamido)butanyl]-His-[Glu14, Arg17, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxy-ten Amidinosyl)butanyl]-His-His-[Glu14, Arg17, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptylamino)butanyl ]-His-[Glu14,His17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-His-[Glu14, Arg17 ,His35]-pramlintide, N-ε-25-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxy] Amino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Ser21, Lys25]-planin, N-ε- 25-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptytylamino)butaninyl]ethoxy} Ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Lys25]-pramlintide, N-ε-21-[2-(2-{2-[ 2-(2-{2-[(S)-4-carboxy-4-(1) 7-carboxyheptadecaneamino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Lys21,Glu35]- Pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[des1, Glu14, His17, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxyundosylamino)butanyl]-[His1, Glu14, His17, His35]-pramlintide, N-α-[(S) 4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1, Glu14, His17, Arg18, His35]-pramlintide, N-α-[(S)-4-carboxy-4 -(19-carboxynonylamino)butanyl]-[Glu14,His17,Arg18,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxyl-19醯Amino)butanyl]-[Glu14,His17,Arg18]-pramlintide, N-ε-17-[2-(2-{2-[2-(2-{2-[(S)-4-carboxyl) 4-(17-carboxyheptadelamido)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Lys17]- Pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Gly-[Glu14,His17,His35]-pramlintide, N-α- [2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptylamino)butaninyl]ethoxy}ethoxy Ethylamino]ethoxy}ethoxy)ethinyl]-[Glu1, Glu14,His17,Pro37]-pramlintide, N-ε-17-[2-(2-{2-[ 2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanosamino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy }Ethoxy)ethinyl]-[Glu14,L Ys17,Ser21,Glu35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[His1,Glu14,His17,Pro37]-Plan Linpeptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Ser-[Glu14,His17,His35]-pramlintide, N-α-[ (S)-4-carboxy-4-(19-carboxyundecylamino)butane -[Glu14,Pro37]-pramlintide, N-ε-1-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14,His17,His35] -Pullinin, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Gly1, Glu14, Arg17, Pro37]-pramlintide, N-α -[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Gly-[His1,Glu14,Arg17,Pro37]-pramlintide, N-α-[(S)- 4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1,Glu14,Arg17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19 -carboxy-nonadesylamino)butanyl]-[Gly1,Glu14,Hi17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino) Butyl]-Gly-[His1, Glu14, His17, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, His35]-pramlintide, N-ε-1-[(S)-4-carboxy-4-(17-carboxyheptadecanosamino)butanyl]-[Glu14,His17,His35]-pramlintide, N -α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Arg1, Glu14, His17, His35]-pramlintide, N-α-[(S)- 4-carboxy-4-(19-carboxynonylamido)butanyl]-[Arg1,Glu14,His17,Pro37]-Planin , N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Arg1,Pro37]-pramlintide, N-α-[(S)-4- Carboxy-4-(19-carboxyundosylamino)butane -[Arg1,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Gly-[Arg1,His35]-Plan Linpeptide, N-α-[(S)-4-carboxy-4-(19-carboxynonanosylamino)butanyl]-Gly-[Arg1, Glu14, His17, His35]-pramlintide, N-α -[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Gly-[Arg1,Glu14,His17,Pro37]-pramlintide, N-α-[(S)- 4-carboxy-4-(19-carboxyundecylamino)butanyl]-Gly-[Arg1,Glu14,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19 -carboxy-nonyl-nonylamino)butanyl]-Gly-[Arg1,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl] -[des1,Glu14,His17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Gly1, Glu14, His17, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Ser1, Glu14, His17, Pro37]-planin, N -α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Ser1, Glu14, His17, His35]-planin, N-α-[(S)- 4-carboxy-4-(19-carboxynonylamido)butanyl]-[des1,Glu14,Pro37]-pramlintide, N- -[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1,Glu14,Pro37]-pramlintide, N-α-[(S)-4-carboxy- 4-(19-carboxynonylideneamino)butane Base]-[His1, Glu14, Ser21, Ser28, Ser29, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[ His1, Glu14, Arg17, Ser21, Ser28, Ser29, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, His17, Arg35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14,His17,Lys35]-pramlintide, N -α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14,His17,His36]-pramlintide, N-α-[(S)-4- Carboxy-4-(19-carboxyundecylamino)butanyl]-[Glu14,His17,His34]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxy-19醯Amino)butanyl]-[Glu14,His17,His32]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, His17, Gln21, His35)-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxy) Amidino) butylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Ser21,His35]-pramlintide, N -α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptylamino)butanylamino]] Oxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[His1, Glu14, His17, Ser21, His35]-planin, N-α-[2-(2 -{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butaninyl]ethoxy}ethoxy)acetamide Ethyloxy}ethoxy) Mercapto]-D-Arg-D-Arg-[Orn1, Glu14, His17, Arg18]-pramlintide, N-α-(19-carboxyundecyl)-Glu-Glu-Arg-[Glu1, Pro37 ]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-Arg-[Glu1,Pro37]-pramlintide, N- --[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Glu-Arg-[Glu1, Glu14, His17, Pro37]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Glu-Arg-[Glu1, Glu14, Arg17, Pro37]-pramlintide, N-α-(19-carboxy-19 Mercapto)-Glu-Glu-Arg-[Glu1, Glu14, His17, Pro37]-pramlintide, N-ε-25-[2-(2-{2-[2-(2-{2-[( S)-4-carboxy-4-(17-carboxyheptadecylamido)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]- [Glu14, Lys25]-pramlintide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxy-ten) Hexamethylamino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Lys21, Ser29, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, His17, Asp21, His35]-pramlintide, N-α-[(S) -4-carboxy-4-(19- Carboxyylammonyl)butanyl]-Glu-[Glu14,His17,Gln21,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)醯丁基]-[Glu14,His17,Gln21,Pro37]-pramlintide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxyl) -4-(19-carboxy seventeen Amidino) butylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Lys21, Ser28, Ser29, His35]-planin , N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadecanium))butanamine Ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,l_ys21,His35]-pramlintide, N-ε-25-[2-(2 -{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadecylamido)butaninyl]ethoxy}ethoxy)acetamide Ethyloxy}ethoxy)ethinyl]-[Glu14,His17,Ser21,Lys25,Ser28,Ser29.His35]-pramlintide, N-ε-25-[2-(2-{2- [2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadecylamido)butaninyl]ethoxy}ethoxy)acetamido]ethoxy Ethyloxy)ethinyl]-[His17,His21,Lys25,Ser28,Ser29.His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxy-19 Amidino)butyric acid]-[Glu14,His17,Gln35,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[ Glu14, His17, Gln21, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Gly1, Glu14, His17, Gln 21, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu1, Glu14, Gln21, Gln35, Pro37] -Pullinin, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14, Gln21, Gln35, Pro37]-pramlintide, N-ε -25-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadecanosamino)butaninyl]ethoxy) }ethoxy)acetamido]ethoxy}ethoxy) Ethyl]-[His17, Lys25, Ser28, Ser29, His35]-pramlintide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-) 4-carboxy-4-(19-carboxyheptadecylamido)butanylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[His17, Lys21, Ser28, His35]-pramlintide, N-ε-25-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyl) Hexamethylamino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, His17, Lys25, Ser28, Ser29, His35] - pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1,Glu14,Arg18,Pro37]-pramlintide, N-ε -21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadecanosamino)butaninyl]ethoxy) }Ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Lys21,Ser28,His35]-pramlintide, N-α-[(S)-4- Carboxy-4-(19-carboxyundosylamino)butanyl]-[Lys11, Glu14, His17, Pro37]-planin, N-α-[(S)-4-carboxy-4-(19-carboxyl 19-nonylamino)butanyl]-[Glu14,His17,Thr21,Pro37]-pramlintide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S) )-4-carboxy-4-(19-carboxyheptadecaneamino) Amidino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Lys21, Ser28, Arg29, His35]-pramlintide, N-ε- 21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptylamino)butanylamino]ethoxy} Ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Lys21,Ser28,Ser29,His35]-pramlintide, N-ε-1-[(S) 4-carboxy-4-(19-carboxynonylamido)butanyl]-Arg-[Glu14, Arg17, His35, Pro37]-pramlintide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptylamino)butanylamino]] Ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Arg18, Lys21, Ser28, Arg29, Arg35]-pramlintide, N-ε-1- [(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14,His17,Gln21,Ala25,Pro26,Ser28,Ser29,Pro37]-pramlintide, N-α- [(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Arg14,Arg17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4 -(19-carboxynonylamido)butanyl]-Gly-[Arg1,Glu14,Arg17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxy-ten Amidinosyl)butanyl]-[Arg17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-Glu- [Arg1, Glu14, Arg17, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-[Arg1, Glu14, Arg17 ,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-Glu-[Arg1, Glu14, Arg17, Gln21, Pro37 ]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-Glu-[Arg1, Glu 14, Arg17, Gln21, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-Glu-[Arg1, Glu14, Arg17, Gln21, Gln31, Gln35, Pro3 7]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-Glu-[Arg1, Glu14, Arg17, Gln21, Gln22, Gln31, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-Arg-[Glu1, Glu14, Arg17, Gln21,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonanosylamino)butanyl]-Glu-Arg-[Glu1, Glu14, Arg17, Gln21, Gln35,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonanosylamino)butanyl]-Glu-Arg-[Glu1, Glu14, Arg17, Gln21, Gln31, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-Arg-[Glu1, Glu14, Arg17, Gln21, Gln22, Gln31, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-Glu-[Glu1, Glu14, Arg17, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, Arg18, Gln21, Ala25, Pro26, Ser28, Ser29, Pro37]-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxy) Amidino) butylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)acetamidine ]-[Glu14,His17,Glu21,Pro37]-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-() 19-carboxy heptadecylamine Butylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Lys21,Pro37]-pramlintide, N-α -[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptytylamino)butaninyl]ethoxy}B Oxy)ethylamino]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Ala21,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4- (19-carboxynonylamido)butanyl]-[Glu14, Arg18, Ser21, Ser22, Ser28, Ser29, Asp31, Asp35, Pro37]-pramlintide, N-α-[(S)-4-carboxy- 4-(19-carboxyundosylamino)butanyl]-[Glu14, Ser21, Ser22, Pro23, Ala25, Pro26, Ser28, Ser29, Asp31, Asp35, Pro37]-pramlintide, N-α-[(S )-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14, Arg17, Asp21, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4- (19-carboxynonylamido)butanyl]-[Glu14, Arg17, Asp35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonanthrylamine Butyl)-[Glu14, Arg17, Asp31, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, Arg17, Asp22, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxylate 19-nonylamino)butanyl]-[Asp3, Glu14, Arg17, Pro37]-pramlintide, N-ε-1-[(S)-4-carboxy-4-(19-carboxyundecylamino)醯醯基]-[Glu14, Arg17, Pro37]-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19) -carboxyheptadecaneamine Butylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Ala21, His35, Pro37]-pramlintide.

38. A pharmaceutical composition comprising a derivative according to any one of paragraphs 1 to 37 and a pharmaceutically acceptable excipient.

39. The pharmaceutical composition of paragraph 38 which is suitable for parenteral administration.

40. A derivative according to any of paragraphs 1 to 37 which is suitable for use as a pharmaceutical.

41. A derivative according to any one of paragraphs 1 to 37, which is useful as a medicament for the treatment or prevention of hyperglycemia, type 2 diabetes, abnormal glucose tolerance, type 1 diabetes, obesity, hypertension Syndrome X, dyslipidemia, cognitive disorders, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, dyspepsia and gastric ulcers.

42. A derivative according to any one of paragraphs 1 to 37 which is useful as a medicament for delaying or preventing progression of type 2 diabetes diseases.

43. A derivative according to any one of paragraphs 1 to 37 which is useful for reducing food intake, reducing beta cell apoptosis, increasing beta-cell function and beta-cell population, and/or restoring beta-cell glucose Sensitive pharmaceuticals.

44. Use of a derivative according to any of paragraphs 1 to 37 for the preparation of a medicament.

45. Use of a derivative according to any one of paragraphs 1 to 37 for the preparation of a medicament for the treatment or prevention of hyperglycemia, type 2 diabetes, abnormal glucose tolerance, type 1 Diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive disorders, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, Indigestion and stomach ulcers.

46. Use of a derivative according to any one of paragraphs 1 to 37 for the manufacture of a medicament for delaying or preventing progression of a type 2 diabetes disease.

47. Use of a derivative according to any one of paragraphs 1 to 37 for the preparation of a medicament for the treatment of obesity, reduction in food intake, reduction of β-cell apoptosis, increase of β-cell function and β-cell population And/or a pharmaceutical for restoring glucose sensitivity of β-cells.

A method of preparing a pharmaceutical composition according to any of paragraphs 38 to 39, which comprises mixing a derivative according to any one of paragraphs 1 to 37 with at least one pharmaceutically acceptable excipient.

49. An amyloid derivative as in the examples.

The invention is further summarized in the following other paragraphs:

An amyloid derivative which is an amyloid analog having up to ten amino acid residue modifications as compared to SEQ ID NO: 2 and which is an alpha-terminal amino acid residue of the amyloid analog a substituent attached to the amine or Lys residue, wherein the substituent comprises an albumin binding moiety, and wherein a. the amino acid residue at position 14 of the amyloid analog is Glu or b. The amyloid is similar The amino acid residue at position 35 is His, Arg, Lys, Asp or Glu or c. The amino acid residue at position 37 of the amyloid analog is Pro.

2. The derivative of paragraph 1, wherein the amino acid residue at position 14 is Glu.

3. A derivative according to paragraphs 1 to 2, wherein the amino acid residue at position 14 is Glu and the amine at position 35 The base acid is His, Arg, Lys, Asp or Glu or d. The amino acid residue at position 14 is Glu and the amino acid at position 37 is Pro.

4. The derivative of paragraphs 1 to 4, wherein the amino acid residue at position 17 is His.

5. The derivative of paragraph 1, wherein the amyloid analog comprises the amino acid sequence of formula 1: Xaa ₁ -Cys-Xaa ₃ -Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu- Ala-Glu-Phe-Leu-Xaa ₁₇ -Xaa ₁₈ -Ser-Ser-Xaa ₂₁ -Xaa ₂₂ -Phe-Gly-Pro-Xaa ₂₆ -Leu-Pro-Pro-Thr-Xaa ₃₁ -Val-Gly-Ser- Xaa ₃₅ -Thr-Xaa ₃₇ Formula (I) (SEQ ID No: 3) wherein Xaa _{1 is} deleted or independently selected from Ala, Cys, Glu, Gly, His, Arg, Ser and Lys; Xaa _{3 is} independently selected from Gly , His, Arg, Ser and Asn; Xaa _{17 is} independently selected from the group consisting of His, Arg, Lys and Val; Xaa _{18 is} independently selected from Arg, Lys and His; Xaa _{21 is} independently selected from the group consisting of Ala, Lys, Gln, Ser and Asn Xaa _{22 is} independently selected from the group consisting of Glu, Gln, Ser, Thr and Asn; Xaa _{26 is} independently selected from the group consisting of Pro, Arg and Ile; Xaa _{31 is} independently selected from the group consisting of Ser, Glu, Asp and Asn; Xaa _{35 is} independently selected from the group consisting of His , Arg, Lys, Asp, and Glu; Xaa _{37 is} independently selected from Pro and Tyr; and wherein the C-terminus may be derivatized as a guanamine.

6. The derivative of paragraphs 1 to 5, wherein the substituent comprises a linkage of an amino acid having from 1 to 10 alpha-amino or Lys residues attached to the N-terminal amino acid residue of the amyloid analog child.

7. The derivative of paragraphs 1 to 6, wherein the linker comprises γGlu.

8. The derivative of paragraphs 1 to 7, wherein the linker is selected from the group consisting of γGlu-C(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -[ NHC(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -] ₁ -NH- and Arg-Arg-γGlu-C(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -[NHC(O)-CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -] ₁ -NH-.

9. The derivative of paragraphs 1 to 8, wherein the derivative comprises a γGlu linker linked to the N-terminus of the amyloid analog and HOOC(CH ₂ ) ₁₈ CO or HOOC (CH ₂ ) as the albumin-binding residue ₁₆ CO-, and wherein the sequence of the amyloid analog comprises: a. Glu at position 14 or His or Arg c at position 17. His at position 35 or Pro at position 37.

10. The derivative of paragraph 1, wherein the derivative is selected from the group consisting of N-α-[(S)-4-carboxy-4-(19-carboxyundecylamino)butanyl]-[ His1, Glu14, His17, Arg18]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[His1, Glu8, Glu14, His17, Arg18]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[des1, Gly3, Glu14, His17, Arg18, Glu31, Gly35] -planin, N-α-[(S)-4-carboxy-4-(19-carboxyundecylamino)butanyl]-[des1, Gly3, Glu14, His17, Arg18, Gln21, Gln22, Glu31, Gly35]-Pullin Peptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu1, Glu14, His17, Arg18]-planin, N-α-[( S)-4-carboxy-4-(19-carboxyundosylamino)butanyl]-[His1, Glu14, His17, Arg18, Ala25, Pro26, Ser28, Ser29]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxyundosylamino)butanyl]-[His1, Glu14, His17, Arg18, Glu25, Ser28, Arg29]-pramlintide, N-α-[(S) 4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1, Glu14, His17, Arg18, Glu22, Ala25, Arg26, Ser28, Ser29]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1, Glu14, His17, Arg18, Ser21, Ser22, Asp31, Asp35]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxyundosylamino)butanyl]-[Glu1, Glu14, His17, Arg18, Pro37]-pramlintide, N-α-[(S)-4-carboxyl -4-(19-carboxynonylamino)butanyl]-His-His-[His1,Glu14,Hi17]-pramlintide, N-α-[(S)-4-carboxy-4-(19- Carboxylamidylamino)butanyl]-His-His-[His1, Glu14, Arg17 ]-pramlintide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxy) Amidino) butylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Arg1, Glu14, His17, Arg18, Lys21]-pramlintide , N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1,Glu14,Arg18]-planin, N-α-[(S)- 4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1, Gly3, Glu14, His17, Arg18, Ser21, Ser22, Glu31, Glu35]-pramlintide, N-α-[(S )-4-carboxy-4-(19-carboxynonylamido)butanyl]-His-[His1,Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4- (19-carboxynonylamido)butanyl]-[His1,Glu14,Hi17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)醯醯基]-His-His-[His1, Glu14, His17, Ala25, Pro26, Ser28, Ser29]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonadecanamide Butyl)-His-His-[His1,Glu14]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-His-His -[His1,Glu14,His17,Arg35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-His-His-[His1, Arg3, Glu14, His17]-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-) 4-(19-carboxyheptadecylamido)butanylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[His1,Glu14, His17, Arg18, Ala19, Thr20, Gln21, Glu22, Leu23]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu1, Gly3, Glu14, Arg18, Ser21, Ser22, Ala25, Pro26, Ser28, Ser29, Glu31 , Arg35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-[Arg1, Glu14, His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-[Glu14,His17]-pramlintide, N-α-[(S)-4 -carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxy-19醯Amino)butanyl]-[des1,Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu1, Glu14 ,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu1, Glu14, His17, Arg18]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-His-His-[His1, Glu14, His17, His35]-pramlintide, N-α- [(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-His-His-[His1,His3,Glu14,His17,His35]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-His-His-[His1,His3,Glu14 ,His17,His29,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonanosylamino)butanyl]-His-His-[His1, Glu14, His17 ,His29,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-His-His-[His1,His3,Glu14,His17]-pramlintide, N-α- [(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-His-His-[His1,Glu14,His17,His29]-pramlintide, N-α-[2-( 2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadecylamido)butaninyl]ethoxy}ethoxy)acetamidine Amino]ethoxy}ethoxy)ethinyl]-D-Arg-D-Arg-[Glu14,His17,Arg18]-pramlintide, N-α-[2-(2-{2-[ 2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadecanosamino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy }Ethoxy)ethinyl]-D-Arg-D-Arg-[Glu14,His17,Arg18,Ala21,Ser35]-pramlintide, N-α-[(S)-4-carboxy-4-( 19-carboxy-nonyl-nonylamino)butanyl]-Ser-Ser-Gly-Ser-Ser-[Gly1, Glu14, His17]-pramlintide, N-α-[(S)-4-carboxy-4-( 19-carboxynonanosylamino)butanyl]-Glu-[His1,Glu14,Hi17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)醯Germanyl]-Glu-Glu-[Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-Glu-[Glu14 ,His17]-pramlintide, N-α- [2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadelamido))butanylamino]ethoxy}ethoxy Ethylamino]ethoxy}ethoxy)ethinyl]-[des1,Glu14,Hi17]-pramlintide, N-α-[(S)-4-carboxy-4-(19- Carboxylamidylamino)butanyl]-Ser-Ser-Gly-Ser-Ser-Gly-[His1,Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-His-His-[His1,Glu14,His17]-pramlintide, N-α-[( S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-[Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(17- Carboxythylideneamino)butanyl]-His-[His1,Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptylamino)butanyl ]-[His1,Glu14,His17]-pramlintide, N-α-{(S)-4-carboxy-4-[(S)-4-carboxy-4-(17-carboxyheptadecaneamino) Butylamine]butyrate}-[des1,Glu14,His17]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14 ,Arg17]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-[Glu14,Arg17]-pramlintide, N-α- [(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14,His17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4 -(19-carboxynonylamido)butanyl]-[Glu14,Arg17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino) ) 醯 醯]]-[Glu10, Glu14, His17]- pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxyundecylamino) butyl -[Glu14,His17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-[Glu14,His17, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14,Arg17,His35]-pramlintide, N-α-[(S)-4 -carboxy-4-(19-carboxyundecylamino)butanyl]-Glu-[Glu14, Arg17, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(17- Carboxythylideneamino)butanyl]-[Glu14,His17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptylamino)butanyl]- Glu-[Glu14,His17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-[Glu14, Arg17, His35]- Pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyheptadelamido)butanyl]-Glu-[Glu14, Arg17, His35]-pramlintide, N-α- [(S)-4-carboxy-4-(19-carboxyundosylamino)butanyl]-His-His-[His1, Glu14, His17, Ser21, Glu35]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14,His17,Ala25,Pro26,Ser28,Ser29,His35]-pramlintide, N-α-[(S) 4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14, Arg17, Ala25, Pro26, Ser28, Ser29, His35]-pramlintide, N-α-[(S)-4 -carboxy-4-(19-carboxyundecylamino)butanyl]-His-[Glu14,His17,His35]- LAN Lin peptide, N-α - [(S) -4- Carboxy-4- (19-carboxynonadecanoylamino acyl amino) butan-acyl] -His-His- [Glu14, His17, His35] - pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-His-[Glu14, Arg17, His35]-pramlintide, N-α-[(S) 4-carboxy-4-(19-carboxynonylamino)butanyl]-His-His-[Glu14, Arg17, His35]-pramlintide, N-α-[(S)-4-carboxy-4 -(17-carboxyheptadecaneamino)butanyl]-His-[Glu14,His17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(17-carboxyl-17醯Amino)butanyl]-His-[Glu14, Arg17, His35]-pramlintide, N-ε-25-[2-(2-{2-[2-(2-{2-[(S)-4) -carboxy-4-(17-carboxyheptadecylamido)butanylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Ser21 , Lys25]-pramlintide, N-ε-25-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxy] Amino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Lys25]-planin, N-ε- 21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanium))butanylamino]ethoxy} Ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Lys21,Glu35]-pramlintide, N-α-[(S)-4-carboxy-4 -(19-carboxynonylamino)butanyl]-[des 1, Glu14, His17, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[His1, Glu14, His17, His35] - pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1, Glu14, His17, Arg18, His35]-pramlintide, N -α-[(S)-4-carboxy-4-(19-carboxyundecylamino)butane -[Glu14,His17,Arg18,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14,His17, Arg18]-pramlintide, N-ε-17-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecane) Butylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,Lys17]-planin, N-α-[(S )-4-carboxy-4-(19-carboxynonylamido)butanyl]-Gly-[Glu14,His17,His35]-planin, N-α-[2-(2-{2-[2 -(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanosamino)butaninyl]ethoxy}ethoxy)ethinyl]ethoxy} Ethoxy)ethinyl]-[Glu1, Glu14,His17,Pro37]-pramlintide, N-ε-17-[2-(2-{2-[2-(2-{2-[(S) )-4-carboxy-4-(17-carboxyheptadecylamido)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[ Glu14, Lys17, Ser21, Glu35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[His1, Glu14, His17, Pro37] - pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Ser-[Glu14,His17,His35]-pramlintide, N-α -[(S)-4-carboxy-4-(19- N-decylamino)butanyl]-[Glu14,Pro37]-pramlintide, N-ε-1-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]- [Glu14, His17, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Gly1, Glu14, Arg17, Pro37]- Pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Gly-[His1,Glu14,Arg17,Pro37]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1,Glu14,Arg17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4 -(19-carboxynonylamino)butanyl]-[Gly1, Glu14, His17, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxyl-19醯Amino)butyric acid]-Gly-[His1,Glu14,His17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]- [Glu14, His35]-pramlintide, N-ε-1-[(S)-4-carboxy-4-(17-carboxyheptadecylamido)butanyl]-[Glu14,His17,His35]-Planin Peptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Arg1, Glu14, His17, His35]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Arg1,Glu14,His17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4 -(19-carboxynonylamino)butanyl]-[Arg1,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl ]-[Arg1,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butyl Mercapto]-Gly-[Arg1,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Gly-[Arg1, Glu14 ,His17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxyundecylamino)butanyl]-Gly-[Arg1, Glu14, His17, Pro37]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxyundecylamino)butanyl]-Gly-[Arg1,Glu14,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4 -(19-carboxynonylamido)butanyl]-Gly-[Arg1,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino) )]][[des1,Glu14,His17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Gly1,Glu14 ,His17,His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Ser1, Glu14, His17, Pro37]-Planin Peptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Ser1, Glu14, His17, His35]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[des1,Glu14,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-( 19-carboxynonylideneamino)butanyl]-[His1,Glu14,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl ]-[His1, Glu14, Ser21, Ser28, Ser29, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19- N-nonylamino)butanyl]-[His1, Glu14, Arg17, Ser21, Ser28, Ser29, His35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxyl Amidoxime -[Glu14,His17,Arg35]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14,His17,Lys35] - pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14,His17,His36]-pramlintide, N-α-[ (S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14,His17,His34]-pramlintide, N-α-[(S)-4-carboxy-4- (19-carboxynonylamido)butanyl]-[Glu14,His17,His32]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)醯醯基]-[Glu14,His17,Gln21,His35)-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-) (19-carboxyheptadecylamido)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Ser21,His35] -Pullinin, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptylamino))) Amino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[His1, Glu14, His17, Ser21, His35]-pramlintide, N-α-[ 2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadelamido))butanylamino]ethoxy}ethoxy Ethylamine ethoxylate }Ethoxy)ethinyl]-D-Arg-D-Arg-[Orn1,Glu14,His17,Arg18]-pramlintide, N-α-(19-carboxyundecyl)-Glu-Glu- Arg-[Glu1, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-Arg-[Glu1,Pro37]- Pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Glu-Arg-[Glu1, Glu14, His17, Pro37]-pramlintide, N-α- [(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Glu-Arg-[Glu1, Glu14, Arg17, Pro37]-pramlintide, N-α-(19-carboxyl 19-mercapto)-Glu-Glu-Arg-[Glu1, Glu14, His17, Pro37]-pramlintide, N-ε-25-[2-(2-{2-[2-(2-{2- [(S)-4-carboxy-4-(17-carboxyheptadecylamido)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethenyl ]-[Glu14,Lys25]-pramlintide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-) Carboxyrytylamino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Lys21, Ser29, His35]-Planin Peptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, His17, Asp21, His35]-pramlintide, N-α-[( S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Glu-[Glu14,His17,Gln21,His35]-pramlintide, N-α-[(S)-4-carboxyl -4-(19-carboxynonylamido)butanyl]-[Glu14,His17,Gln21,Pro37]-pramlintide, N-ε-21-[2-(2-{2-[2-(2) -{2-[(S)-4-carboxy-4-(19- Carboxythylidene amino) butylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Lys21, Ser28, Ser29, His35]- Pramlintide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptylamino)) Amidino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Lys21,His35]-pramlintide, N-ε-25-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptylamino)butanylamino]] Ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Ser21,Lys25,Ser28,Ser29.His35]-pramlintide, N-ε- 25-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptytylamino)butanylamino]ethoxy} Ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[His17,His21,Lys25,Ser28,Ser29.His35]-pramlintide, N-α-[(S)-4 -carboxy-4-(19-carboxyundecylamino)butanyl]-[Glu14,His17,Gln35,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19- Carboxylamidinyl)butanyl]-[Glu14,His17,Gln21,Gln35,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino) )]][Gly1, Glu14, His17, Gln21, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]- [Glu1, Glu14, Gln21, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, Gln21, Gln35 ,Pro37]-pramlintide, N-ε-25-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxy] Amino)butynylamino] Ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[His17, Lys25, Ser28, Ser29, His35]-pramlintide, N-ε-21-[2 -(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptylamino)butaninyl]ethoxy}ethoxy) Ethylamino]ethoxy}ethoxy)ethinyl]-[His17,Lys21,Ser28,His35]-pramlintide, N-ε-25-[2-(2-{2-[2- (2-{2-[(S)-4-carboxy-4-(19-carboxy seventeen) Amidino) butylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Lys25,Ser28,Ser29,His35]-Plan Linpeptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[His1,Glu14,Arg18,Pro37]-pramlintide, N-ε-21 -[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptytylamino)butaninyl]ethoxy}B Oxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Lys21,Ser28,His35]-pramlintide, N-α-[(S)-4-carboxy- 4-(19-carboxynonylamido)butanyl]-[Lys11,Glu14,His17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxy-19 Amidino) butyl sulfhydryl]-[Glu14,His17,Thr21,Pro37]-pramlintide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-) 4-carboxy-4-(19-carboxyheptadecylamido)butanylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Lys21, Ser28, Arg29, His35]-pramlintide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19) -carboxyheptadecaneamino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Lys21,Ser28,Ser29, His3 5]-pramlintide, N-ε-1-[(S)-4-carboxy-4-(19-carboxynonanosylamino)butanyl]-Arg-[Glu14, Arg17, His35, Pro37]-Plan Lin peptide, N-ε-21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadecanoyl))) Amino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Arg18, Lys21, Ser28, Arg29, Arg35]-pramlintide, N-ε 1-[(S)-4-carboxy-4-(19-carboxynonylamido)butane Base]-[Glu14,His17,Gln21,Ala25,Pro26,Ser28,Ser29,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino) Butyl]-[Arg14, Arg17, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Gly-[Arg1, Glu14, Arg17,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Arg17,Pro37]-pramlintide, N-α -[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Glu-Glu-[Arg1,Glu14,Arg17,Pro37]-pramlintide, N-α-[(S )-4-carboxy-4-(19-carboxyundecylamino)butanyl]-Glu-[Arg1,Glu14,Arg17,Pro37]-pramlintide, N-α-[(S)-4-carboxy- 4-(19-carboxynonylamido)butanyl]-Glu-Glu-[Arg1, Glu14, Arg17, Gln21, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4- (19-carboxynonylamido)butanyl]-Glu-Glu-[Arg1, Glu14, Arg17, Gln21, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4- (19-carboxynonylamido)butanyl]-Glu-Glu-[Arg1, Glu14, Arg17, Gln21, Gln31, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy- 4-(19-carboxynonylamido)butanyl]-Glu-Glu-[Arg1, Glu14, Arg17, Gl N21, Gln22, Gln31, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylideneamino)butane ]]-Glu-Arg-[Glu1, Glu14, Arg17, Gln21, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl] -Glu-Arg-[Glu1, Glu14, Arg17, Gln21, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl] -Glu-Arg-[Glu1, Glu14, Arg17, Gln21, Gln31, Gln35, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino) Butyl]-Glu-Arg-[Glu1, Glu14, Arg17, Gln21, Gln22, Gln31, Gln35, Pro37]-Pullin, N-α-[(S)-4-carboxy-4-(19-carboxy-19 Amidino)butyric acid]-Glu-Glu-[Glu1, Glu14, Arg17, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)丁醯基]-[Glu14, Arg18, Gln21, Ala25, Pro26, Ser28, Ser29, Pro37]-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S) )-4-carboxy-4-(19-carboxyheptadecylamido)butanylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[ Glu14, His17, Glu21, Pro37]-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyl) Hexamethylamino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Ly S21,Pro37]-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxyheptadecane) Butylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14,His17,Ala21,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14, Arg18, Ser21, Ser22, Ser28, Ser29, Asp31, Asp35, Pro37]-Pullin Peptide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, Ser21, Ser22, Pro23, Ala25, Pro26, Ser28, Ser29, Asp31, Asp35 ,Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14, Arg17, Asp21, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14, Arg17, Asp35, Pro37]-pramlintide, N-α-[(S) 4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14, Arg17, Asp31, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-( 19-carboxynonylamidoalkyl)butanyl]-[Glu14, Arg17, Asp22, Pro37]-pramlintide, N-α-[(S)-4-carboxy-4-(19-carboxynonylamino)醯]][[Asp3,Glu14,Arg17,Pro37]-pramlintide, N-ε-1-[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14 ,Arg17,Pro37]-pramlintide, N-α-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxy] Amino)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl] - [Glu14, Ala21, His35, Pro37] - pramlintide.

A pharmaceutical composition comprising a derivative according to any one of paragraphs 1 to 10 and a pharmaceutically acceptable excipient.

12. A derivative according to any one of paragraphs 1 to 10 which is suitable for use as a pharmaceutical.

13. A derivative according to any one of paragraphs 1 to 10 which is useful for treating obesity, reducing food intake, reducing β-cell apoptosis, increasing β-cell function and β-cell population and/or restoring β- A glucose-sensitive drug for cells.

14. Use of a derivative according to any of paragraphs 1 to 10 for the preparation of a medicament.

15. A method of preparing a pharmaceutical composition comprising admixing a derivative of any of paragraphs 1 to 10 with at least one pharmaceutically acceptable excipient.

The invention will now be described, by way of example only.

Example

The prepared multi-peptides are shown in Table 10.

The in vitro efficacy of the multi-peptide on amyloid receptors and calcitonin receptors from rats and humans (as described in assay (II)) is shown in Table 11.

Polypeptide solubility was tested as described in assay (IV) and the results are shown in Table 12.

The physical stability of the amyloid derivative was tested in the ThT assay (assay (III)) and the data is given in Table 13.

The effect of the amyloid derivative was tested in the food intake assay (assay (I)) and the results are shown in Table 14.

The half-life of the amyloid derivatives of the invention was tested in piglets as described in assay (IX) and the information is given in Table 15.

abbreviation

Some of the abbreviations used in the examples are as follows: Acm : acetaminomethyl

HATU : (O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl hexafluorophosphate )

HBTU : 2-(IH-benzotriazol-1-yl-)-1,1,3,3-tetramethyl hexafluorophosphate

Fmoc : 9H-茀-9-ylmethoxycarbonyl

Boc : third butoxycarbonyl

Mtt : 4-methyltrityl

DCM : dichloromethane

TIPS : triisopropyl decane

TFA : trifluoroacetic acid

NMP : 1-methyl-pyrrolidin-2-one

HOAt : 1-hydroxy-7-azabenzotriazole

DIC : diisopropylcarbodiimide

Trt : trityl

Verification

In the following examples, the following assays were involved: assay (I) - experimental protocol for testing the efficacy of appetite using a rat model of ad libitum feeding

Verification (II)a - Functional assay - human calcitonin and amyloid receptor assay

Verification (II)b - Functional assay - rat calcitonin and rat amyloid receptor assay

Verification (III) - ThT fibrillation assay for assessing the physical stability of protein formulations

Test (IV) - Determination of Solubility

Assay (V) - Determination of binding to human amyloid receptors

Test (VI) - Determination of binding to rat amyloid receptor

Assay (VII) - Determination of binding to human calcitonin receptor

Assay (VIII) - Determination of binding to rat calcitonin receptor

Test (IX) -pK-measure T1⁄2 of piglets

Assay (X) -pK-determination of T1⁄2 in rats

Assay (I) - An experimental protocol for testing the efficacy of appetite using a rat model that is fed ad libitum

Experiments were performed using Sprague Dawley (SD) rats from Taconic Europe, Denmark. The rats had a body weight of 200-250 g at the start of the experiment. The experiment was started after the rats arrived at least 10-14 days in order to adapt to the experimental environment. During this time, the animals were treated at least 2 times. Upon arrival, the rats were individually housed for one week to house the light/dark period (meaning to turn off the lights during the day and light up at night) for two weeks. Since the rats were normally active during the dark period and were fed a large portion of their daily food intake, the rats were given medication before the lights were turned off in the morning. This configuration produces the lowest data changes and the highest test sensitivity. Experiments were performed in rat cages and rats were free to ingest food and water during the adaptation period and during the experiment. Each dose of the derivative was tested in a group of 5-8 rats. A vehicle group of 6-8 rats was included in each group of tests.

The rats were administered once per body weight (ipally (ip), orally (po) or subcutaneously (sc) with a solution of 0.01-3 mg/kg). The administration time of each group was recorded. After administration, the rats were returned to their cages, in which the rats were then given food and water. Food consumption was recorded continuously, individually, by online registration or manually, for 7 hours, and then recorded after 24 hours and sometimes 48 hours. At the end of the experimental period, the animals were euthanized.

Individual data is recorded in the Microsoft excel form. After applying the Grubbs statistical evaluation test to the outliers, the outliers were excluded and the results were graphically presented using the GraphPad Prism program.

Verification (II) - Functional Verification

Test (II) a-human calcitonin and amyloid receptor assay

1. Overview of luciferase assay

Activation of the calcitonin receptor and the amyloid receptor (a total expression of calcitonin receptor and receptor activity regulatory protein (RAMP)) increases intracellular cAMP concentration. Thus, transcription is activated by a promoter containing multiple copies of a cAMP response element (CRE). Thus, amylase can be measured by using a CRE luciferase reporter gene introduced into BHK cells which also exhibit calcitonin receptor or amyloid receptor. active.

2. Construction of calcitonin (a) receptor and amyloid 3 (a) receptor / CRE-luc cell line.

The BHK570 cell line was stably transfected with the human calcitonin receptor and CRE-reactive luciferase reporter gene. Cell lines were further transfected with RAMP-3 using standard methods. This causes the calcitonin receptor to become an amyloid 3 (a) receptor. Methotrexate, neomycin, and hygromycin are selectable markers for luciferase, calcitonin receptor, and RAMP-3, respectively.

3. Luciferase assay

For activity assays, BHK calcitonin (a) receptor/CRE-luc cells or amyloid 3 (a) receptor/CRE-luc cells were seeded at a density of approximately 20.000 cells/well in white 96-well plates. in. Cells were stored in 100 μl of growth medium (DMEM with 10% FBS, 1% Pen/Strep, 1 mM sodium pyruvate, 250 nM methotrexate, 500 μg/ml neomycin, and 400 μg/ml hygromycin) in. After incubated overnight at 37 [deg.] C and CO ₂ 5%, from 50 [mu] l / well assay medium (DMEM (without phenol red), Glutamax ^TM, 10% FBS and 10 mM Hepes, pH 7.4) replacing the growth medium. In addition, add 50 μl/well of assay buffer containing the standard or sample. After incubation for 3 hours at 37 ° C and 5% CO ₂ , the assay medium with standards or samples was removed and replaced with 100 μL/well PBS. Furthermore, addition of 100 [mu] l / well LucLite ^TM. The plates were sealed and incubated for 30 minutes at room temperature. Finally, the luminescence was measured in a SPC (single photon counting) mode on a TopCounter (Packard).

Characterization of (II) b-rat calcitonin and rat amyloid receptor assay

cAMP verification overview

Activation of the calcitonin receptor and the amyloid receptor (a total expression of calcitonin receptor and receptor activity regulatory protein (RAMP)) increases intracellular cAMP concentration. To quantify cAMP levels in transiently transfected cells, the use of Adenylyl Perkin Elmer Cyclase Activation FlashPlate ^® assay. The rationale for the FlashPlate ^® assay is the competition between a radioactive cAMP produced by a cell and a non-radioactive cAMP for a fixed number of binding sites.

Rat calcitonin (a) receptor cells and rat amyloid 3 (a) receptor cells were constructed.

Transient transfer of rat calcitonin (a) receptor or amyloid 3 (a) receptor (rat calcitonin (a) receptor + rat RAMP3) using FuGENE ^® 6 (Roche) according to the manufacturer's recommendations Dye BHK tk'ts 13 cells.

cAMP check

24 hours after transient transfection, cells (rat calcitonin (a) receptor cells or rat amyloid 3 (a) recipient cells) (100.000 cells/well) were added to have FlashPlate stimulation buffer and IBMX Samples or standards in 96-well FlashPlates ^® and incubated for 30 minutes. The mixture is formed to detect the manufacturer's protocol and TopCounter on ^TM (Packard) scintillation measurement in after 3 hours of incubation.

Test (III) - ThT fibrillation assay for assessing the physical stability of protein formulations

The low physical stability of the multi-peptide may cause the formation of amyloid fibrils (the well-ordered fibrillar macromolecular structure in the sample is observed), eventually leading to gel formation. Physical stability has traditionally been checked by visual inspection of samples. Measure. However, these measurements are very subjective and depend on the observer.

Therefore, the use of small molecule indicator probes is more advantageous. Thioflavin T (ThT) is the probe and has a significant fluorescent label when bound to fibrils [Naiki et al. (1989) Anal. Biochem. 177 , 244-249; LeVine (1999) Methods. Enzymol. 09 , 274-284].

The time course of fibril formation can be described by a sigmoidal curve having the following expression [Nielsen et al. (2001) Biochemistry 40 , 6036-6046]:

Here, F is the ThT fluorescence at time t. The constant t ₀ is the time required to reach 50% of the maximum fluorescence. Two important parameters describing the formation of fibrils are the lag time calculated from t ₀ -2τ and the apparent rate constant K _app =1/τ.

It is believed that the formation of a partially folded multi-peptide intermediate is a general initiation mechanism for fibrillation. They are rarely nucleated to form a template on which other intermediates can be assembled and continue to form fibrils. The lag time corresponds to the time interval at which the critical core mass is formed, and the apparent rate constant is the rate at which the fibrils themselves are formed.

Sample Preparation

Fresh samples were prepared prior to each assay. Each sample composition is described in each specific example. The pH of the sample was adjusted to the desired value using an appropriate amount of concentrated NaOH and HClO ₄ or HCl. A sample of Thioflavin T was added to a stock solution to a final concentration of 1 μM.

The sample was placed in a 200 μl aliquot of a 96-well microtiter plate (Packard OptiPlate ^TM -96, white polystyrene) in. Typically, four or eight replicates of each sample are placed in a row of wells (corresponding to one test condition). The plate was sealed with a Scotch Pad (Qiagen).

Cultivation and fluorescence measurement

Incubation, shaking and fluorescence emission measurements at the specified temperatures were performed in a Fluoroskan Ascent FL fluorescence plate reader or a Varioskan plate reader (Thermo Labsystems). Adjust the temperature to 37 °C. In all the data presented, the orbital oscillation was adjusted to 960 rpm with an amplitude of 1 mm. Fluorescence measurements were performed using excitation light through a 444 nm filter and emission measurements through a 485 nm filter.

Each operation was started by incubating the plate for 10 minutes at the assay temperature. The plate was measured every 20 minutes for a predetermined time. Between each measurement, the plate was shaken and heated as described.

Data processing

Measurement points were saved in Microsoft Excel format for further processing and curves were drawn and fitted using GraphPad Prism. In the absence of fibrils, the background emission from ThT is negligible. The data points are typically the average of four or eight samples and are displayed using standard deviation error bars. Only the data obtained in the same experiment (i.e., the sample on the same plate) are presented in the same figure to ensure that a relative measure of fibrillation between experiments is obtained.

The data set can be fitted to equation (1). However, since the full S-curve is not always obtained in the measurement time in this case, the degree of fibrillation is expressed as ThT fluorescence displayed as a sample mean value at different time points and displayed in standard deviation.

Measuring initial concentration and final concentration

The multi-peptide concentration of each test formulation was measured before the application of the ThT fibrillation assay ("initial") and after the completion of the ThT fibrillation ("ThT assay"). The concentration was determined by reverse phase HPLC using a pramlintide standard as a reference. After the completion of the measurement, 150 μl was collected from each replicate and transferred to an Eppendorf tube. The replicates were centrifuged at 30,000 G for 40 minutes. The supernatant was filtered through a 0.22 μm filter before application to the HPLC system.

Test (IV) - Determination of Solubility

Polypeptide is dissolved in water at about 500 nmol/ml with a series of buffers (100 mM glycine glycine pH 3.0, 100 mM glycine glycine pH 4.0, 100 mM glycine glycine pH 5.0, 100 mM 1,3-bis[tris(hydroxymethyl)methylamino]propane pH 6.0, 100 mM 1,3-bis[tris(hydroxymethyl)methylamino]propane pH 6.5, 100 mM 1, 3-bis[tris(hydroxymethyl)methylamino]propane pH 7.0, 100 mM 1,3-bis[tris(hydroxymethyl)methylamino]propane pH 7.5, 100 mM 1,3-two [three (three Hydroxymethyl)methylamino]propane pH 8.0) 1:1 mixed. After 18 hours, the samples were centrifuged at room temperature and the multi-peptide concentration was determined by UPLC.

Assay (V) - Determination of binding to human amyloid receptors

Binding assays were performed using PerkinElmer's Scintillation Proximity Assay (SPA) beads (RPNQ0001) and cell membranes from amyloid 3(a)/CRE-luc cells (as described in assay (II)) were used. Cell membranes were prepared in the following manner; cells were washed with PBS and incubated with Versene for approximately 5 minutes prior to collection. The cells were washed with PBS and the cell suspension was centrifuged at 1000 rpm for 5 minutes. The cells were homogenized (ultrathurrax) in a buffer containing 20 mM Na-HEPES and 10 mM EDTA (pH 7.4) and centrifuged at 20.000 rpm for 15 minutes. The resulting pellet was resuspended, homogenized and centrifuged (20.000 rpm, 15 minutes) in a buffer (pH 7.4, buffer 2) containing 20 mM Na-HEPES and 0.1 mM EDTA. The resulting pellet was resuspended in Buffer 2 and the protein concentration was measured (BCA Protein Assay, Pierce). The homogenate remained cold throughout the procedure. The cell membrane was kept at -80 °C until use. The assay was performed in a total volume of 40 μl in a 384-well Optiplate (PerkinElmer). The cell membrane is mixed with SPA beads. The final concentration of the cell membrane was 35 ng/μL and the SPA beads were 0.05 mg/well. Test compounds were dissolved in DMSO and further diluted in assay buffer (50 mM Hepes, pH 7.4, 1 mM CaCl ₂ , 5 mM MgCl ₂ , 0.1% OA, and 0.02% Tween 20). The radioligand ¹²⁵ I-rat amylase (NEX 448 PerkinElmer) was dissolved in assay buffer and added to the Optiplate at a final concentration of 50 pM/well (about 20.000 cpm/10 μl). The final mixture was incubated for 120 minutes at 25 ° C with shaking at 400 rpm before centrifugation (1500 rpm, 10 minutes). Samples were analyzed on a TopCounter ^TM (Packard). The IC ₅₀ value was calculated as the receptor affinity strength using (Panel point binding competition analysis) GraphPad Prism5.

Test (VI) - Determination of binding to rat amyloid receptor

Except as described above using the cell membrane prepared by transiently transfecting rat calcitonin receptor rat RAMP 3 BHK tk'ts 13 cells with the molar ratio (1:2), as described above (assay (V)-determination) Characterization with binding to human amyloid receptors). BHK tk'ts 13 cells were transiently transfected with rat calcitonin receptor using FuGENE® 6 (Roche) according to the manufacturer's recommendations. Cells were grown in DMEM with 10% FBS and 1% Pen/Strep. About 48 hours after transfection At the time, the cells were collected and a cell membrane was prepared.

Assay (VII) - Determination of binding to human calcitonin receptor

Binding assays were performed using PerkinElmer's Scintillation Proximity Assay (SPA) beads (RPNQ0001), and cell membranes prepared from the BHK tk'ts 13 cell line were stably transfected with human calcitonin receptor and CRE-reactive luciferase. dye. Cell membranes were prepared in the following manner; cells were washed with PBS and incubated with Weil for approximately 5 minutes prior to collection. The cells were washed with PBS and the cell suspension was centrifuged at 1000 rpm for 5 minutes. The cells were homogenized (ultrathurrax) in a buffer containing 20 mM Na-HEPES and 10 mM EDTA (pH 7.4) and centrifuged at 20.000 rpm for 15 minutes. The resulting pellet was resuspended, homogenized and centrifuged (20.000 rpm, 15 minutes) in a buffer (pH 7.4, buffer 2) containing 20 mM Na-HEPES and 0.1 mM EDTA. The resulting pellet was resuspended in Buffer 2 and the protein concentration was measured (BCA Protein Assay, Pierce). The homogenate remained cold throughout the procedure. The cell membrane was kept at -80 °C until use. The assay was performed in a total volume of 40 μl in a 384-well Optiplate (PerkinElmer). The cell membrane is mixed with SPA beads. The final concentration of cell membrane was 35 ng/μL and the final concentration of SPA beads was 0.05 mg/well. Test compounds were dissolved in DMSO and further diluted in assay buffer (50 mM Hepes, pH 7.4, 1 mM CaCl ₂ , 5 mM MgCl ₂ , 0.1% OA, and 0.02% Tween 20). The radioligand ¹²⁵ I-calcitonin (NEX422 PerkinElmer) was dissolved in assay buffer and added to the Optiplate at a final concentration of 75 pM/well (about 30.000 cpm/10 μl). The final mixture was incubated for 120 minutes at 25 ° C with shaking at 400 rpm before centrifugation (1500 rpm, 10 minutes). Samples were analyzed on a TopCounter ^TM (Packard). The IC ₅₀ value was calculated as the receptor affinity strength using (Panel point binding competition analysis) GraphPad Prism5.

Assay (VIII) - Determination of binding to rat calcitonin receptor

The assay was performed as described above (assay (VII) - assay binding to human calcitonin receptor), except that the cell membrane prepared from BHK tk'ts 13 cells transiently transfected with the calcitonin receptor was used. BHK tk'ts 13 cells were transiently transfected with rat calcitonin receptor using FuGENE® 6 (Roche) according to the manufacturer's recommendations. Cells were grown in DMEM with 10% FBS and 1% Pen/Strep. About 48 hours after transfection, cells were harvested and cell membranes were prepared.

Determination (IX)-pK-determination of T1⁄2 in piglets

T1⁄2 is the terminal half-life of the compound in plasma = In2 / λ _z . λ _z is the first-order rate constant associated with the terminal (log linear) portion of the plasma concentration-time curve and is estimated from linear regression of time and log concentration.

The T1⁄2 value of the amyloid analogs of the invention was determined using pharmacokinetic studies of male Gottingen piglets from Ellegaard Göttingen Minipigs ApS and followed the principles of experimental animal care.

Approximately 6-10 days of acclimation is allowed before the animal enters the study. At the beginning of the acclimation period, the piglets are approximately 5 to 12 months old and are in the range of 7-35 kg. Two central venous catheters were inserted into the piglets for blood collection.

The study was carried out in an animal room where the illumination was about 12 hours bright and 12 hours dark. Animals are individually housed.

Animals were free to obtain household quality drinking water during the study, but were typically fasted overnight before dosing until about 6-12 hours after dosing. animal Weighed on arrival and on the dosing day.

In the study of the present invention, the test substance was administered subcutaneously at a dose of about 2 nmol/kg. Animals received a single subcutaneous injection. Subcutaneous injection was performed on the right side of the neck about 5-7 cm from the ear and 7-9 cm from the center of the neck. There is a retaining ring on the needle during injection, allowing the needle to enter approximately 0.5 cm. The animals to which each test substance is administered are typically three, but in some cases two or four.

The complete plasma concentration-time characteristics of each animal were obtained (using 12-16 sampling points). In the examples, blood samples were collected according to the following schedule: after subcutaneous administration: before administration (0), after injection 0.5, 1, 2, 4, 6, 8, 12, 24, 48, 72, 96, 120 168 and 240 hours.

In some cases, other blood samples up to 288 hours after injection were also obtained.

0.5 to 2 ml of blood was drawn from each animal at each sampling time. Blood samples were taken via a central venous catheter.

Blood samples were collected into EDTA tubes (ie, Sarstedt Micro tubes 1.3 mL K3E). Blood samples were placed on ice for up to 20 minutes prior to centrifugation. Plasma was separated using centrifugation (i.e., at 4 ° C, 10 minutes, 1500 G) and immediately transferred to a Micronic tube. Approximately 200 μl of plasma was transferred to each Micronic tube. Plasma was stored at -20 ° C until assay. The amyloid content of the plasma samples was assayed using an ELISA assay.

Plasma concentration-time characteristics were analyzed using non-compartmental pharmacokinetic analysis (NCA) using WinNonlin Professional 5.0 (Pharsight, Inc., Mountain View, CA, USA). NCA was performed using individual plasma concentration-time characteristics of each animal. T1⁄2 is the terminal half-life = In2/λ _z and is determined by λ _z ; λ _z is the first-order rate constant associated with the terminal (log-linear) portion of the curve and is estimated by linear regression of time and log concentration.

ELISA plasma assay for the determination of amylase

The human amyloid ELISA is a sandwich immunoassay based on monoclonal antibodies for determining amylose content in human plasma. The capture antibody recognizes 1-20 fragments of human amyloid, amyloid (deaminyl amyloid), amyloid, but does not recognize reduced amyloid. The detection antibody binds to reduced or unreduced human amylase, but does not bind to amyloid and is complexed with streptavidin-alkaline phosphatase. The substrate 4-methylcoumarin phosphate was applied to the intact sandwich and the fluorescent signal monitored at 355 nm/460 nm was proportional to the amount of amyltin present in the sample.

MS method for quantifying amylose

40 μl of plasma was diluted with 120 μl of 66.67% EtOH + 1% HCOOH and mixed. Centrifuge at 13,000 rpm for 20 minutes at 4 °C. Supernatants were analyzed by LC-MS method on a Sciex API 3000 and quantified using standards prepared in plasma.

Assay (X)-pK-determination of T1⁄2 in rats

T1⁄2 is the terminal half-life of the compound in plasma = In2 / λ _z . λ _z is the first-order rate constant associated with the terminal (log linear) portion of the plasma concentration-time curve and is estimated from a linear regression of time and log concentration.

The T1⁄2 value of the amyloid analogs of the invention was determined by pharmacokinetic studies from Spodogley male rats of Taconic Europe and followed the principles of experimental animal care.

A 7-day acclimation period is allowed before the animals enter the study. At the beginning of the acclimation period, the rats were in the range of 300-400 g body weight. A permanent catheter for blood collection is inserted into the rat carotid artery.

The study was carried out in an animal room where the illumination was about 12 hours bright and 12 hours dark. Animals are individually housed and free to access food and water due to the catheter. Animals were weighed on the dosing day.

In the study of the present invention, the test substance was administered subcutaneously at a dose of about 20 nmol/kg. Animals received a single subcutaneous injection of the neck of a syringe using a 25G needle. The animals to which the various test substances are administered are typically three, but in some cases two or four.

Complete plasma concentration-time characteristics of each animal were obtained (8-10 sampling points were used). In the examples, blood samples were collected according to the following schedule: after subcutaneous administration: before administration (0), 0.5, 1, 1.5, 2, 4, 6, 12, 24, 48, and 72 hours after injection.

0.08 to 0.10 ml of blood was drawn from each animal at each sampling time. A blood sample is taken via a catheter.

Blood samples were collected into EDTA tubes. Blood samples were placed on ice for up to 20 minutes prior to centrifugation. Plasma was separated using centrifugation (i.e., at 4 ° C, 10 min, 1500 G) and immediately transferred to a Micronic tube or PCR plate.

Transfer approximately 40 μl of plasma and store at -20 °C until assay. The amyloid content of the plasma samples was assayed using an ELISA assay.

Plasma concentration-time characteristics were analyzed using non-compartmental pharmacokinetic analysis (NCA) using WinNonlin Professional 5.0 (Pharsight, Inc., Mountain View, CA, USA). NCA was performed using individual plasma concentration-time characteristics of each animal. T1⁄2 is the terminal half-life = In2/λ _z and is determined by λ _z ; λ _z is the first-order rate constant associated with the terminal (log-linear) portion of the curve and is estimated by linear regression of time and log concentration.

preparation

The multi-peptide sequence was prepared according to the multi-peptide synthesis mentioned below and the compounds provided in the table (for example, Table 10) were prepared according to the synthesis mentioned below.

A multi-peptide synthesis method was carried out by Fmoc chemistry on a microwave-based Liberty multi-peptide synthesizer (CEM Corporation, North Carolina). The resin is a Tentagel S RAM with a loading of about 0.25 mmol/g or a PAL-ChemMatrix with a loading of about 0.43 mmol/g. The coupling chemistry was DIC/HOAt in NMP using a 0.3 M amino acid solution in NMP and a 6-8 molar excess. Coupling conditions are 5 minutes at up to 70 °C. Deprotection is the use of NMP containing 5% piperidine up to 70 °C. The protected amino acid used was a standard Fmoc-amino acid (supplied by, for example, Anaspec or Novabiochem) dissolved in 0.3 M HOAt in NMP at 0.3 M.

Another multi-peptide synthesis method was performed by Fmoc chemistry on a Prelude multi-peptide synthesizer (Protein Technologies, Arizona). The resin is a Tentagel S RAM with a loading of about 0.25 mmol/g or a PAL-ChemMatrix with a loading of about 0.43 mmol/g. The coupling chemistry was DIC/HOAt in NMP using a 0.3 M amino acid solution in NMP and a 6-8 molar excess. Coupling conditions are single or double coupling at room temperature for 1 or 2 hours. Deprotection using NMP containing 20% piperidine. The protected amino acid used was a standard Fmoc-amino group dissolved in 0.3 M of NMP containing 0.3 M HOAt. Acid (supplied by, for example, Anaspec or Novabiochem).

Another method for multi-peptide synthesis is to use the FastMoc UV protocol supplied by the manufacturer on a 0.25 mmol or 1.0 mmol scale Applied Biosystems 433 multi-peptide synthesizer, which is mediated by HBTU or HATU in NMP. Coupling, and UV monitoring of the Fmoc protecting group for deprotection. The starting resin for the synthesis of the multi-peptide guanamine is Rink-Amide resin. The protected amino acid derivative used is a standard Fmoc-amino acid (supplied by, for example, Anaspec or Novabiochem) suitable for use in a pre-weighed cartridge of the ABI 433A synthesizer.

When it is desired to chemically modify the amine acid side chain, the amine acid is incorporated as Lys (Mtt), and the N-terminal amino acid is incorporated as a Boc-amino acid, or if the N-terminal amino acid is used as Fmoc- Upon incorporation of the amino acid, the Fmoc group was removed and the N-terminus was protected by treatment with NMP containing 6 equivalents of Boc-carbonate and 6 equivalents of DIPEA for 30 minutes. The resin was washed with NMP and DCM and the Mtt group was removed by suspending the resin in pure hexafluoroisopropanol for 20 minutes followed by washing with DCM and NMP. The lysine is chemically modified as follows: by performing one or more automated steps on Liberty or ABI 433 or performing one or more manual coupling steps at room temperature by synthesizing the same with the peptide. Add one or more of the following building blocks. After synthesis, the resin was washed with DCM and dried, and the peptide was cleaved from the resin by TFA/TIPS/water (92.5/5/2.5 or 95/2.5/2.5) for 2 hours, followed by precipitation with 4 volumes of diethyl ether. It was further washed with diethyl ether and dried. If the polypeptide contains cysteine protected with an Acm group, the polypeptide is redissolved in water at 2-5 mg/ml, the pH is adjusted to below 4, and by using 4 equivalents of iodine ( 2% The w/v was treated in methanol for 15 minutes to form a disulfide bridge. Alternatively, a disulfide bridge is formed on the resin by using Trt as a protecting group for cysteine and treating with NMP containing 10 equivalents of iodine for 1 hour. In this case, the crude polypeptide was directly purified after cleavage and diethyl ether precipitation.

Purification: Crude multipeptide was purified by semi-preparative HPLC on a 20 mm x 250 mm column packed with 5 μ or 7 μC-18 cerium oxide. The multi-peptide solution was pumped onto the HPLC column and the precipitated peptide was dissolved in 5 ml of 50% aqueous acetic acid and diluted to 20 ml with H ₂ O and injected onto the column, followed by 50 at 40 ° C. It was eluted with a gradient of 40-60% CH ₃ CN/0.1% TFA 10 mL/min during the minute. The fractions containing the multi-peptide were collected. The purified multi-peptide was lyophilized after diluting the eluate with water.

For analysis of HPLC fractions and final products, RP-HPLC analysis was performed using UV detection at 214 nm and, for example, Vydac 218TP54 4.6 mm x 250 mm 5 μ C-18 cerium oxide column (The Separations Group, Hesperia, USA) and at 42 Dissolve at, for example, 1 mL/min at °C. One of the four different dissolution conditions is most often used: A1: The column is equilibrated with a buffer consisting of 0.1 M(NH ₄ ) ₂ SO ₄ , which is adjusted to pH 2.5 with concentrated H ₂ SO ₄ and at 50 min. During this time, it was dissolved by a gradient of 0% to 60% CH ₃ CN in the same buffer.

B1: Equilibrate the column with 0.1% TFA/H ₂ O and elute from 0% CH ₃ CN/0.1% TFA/H ₂ O to 60% CH ₃ CN/0.1% TFA/H ₂ O over a period of 50 minutes. .

B6: Equilibrate the column with 0.1% TFA/H ₂ O and elute from 0% CH ₃ CN/0.1% TFA/H ₂ O to 90% CH ₃ CN/0.1% TFA/H ₂ O in 50 minutes. .

Alternatively, RP-HPLC analysis was performed using UV detection at 214 nm and Symmetry 300, 3.6 mm x 150 mm, 3.5 μC-18 erbium dioxide column (Waters) dissolved at 1 mL/min at 42 °C.

B4: Equilibrate the column with 0.05% TFA/H ₂ O and elute from 5% CH ₃ CN/0.05% TFA/H ₂ O to 95% CH ₃ CN/0.05% TFA/H ₂ O during 15 minutes. .

The identity of the multi-peptide was confirmed by MALDI-MS on a Bruker Microflex.

The prepared multi-peptides are shown in Table 10 (provided previously): Observations

All references (including publications, patent applications, and patents) cited herein are hereby incorporated by reference in their entirety in their entirety in the extent of the extent of The disclosure is hereby incorporated by reference in its entirety herein in its entirety herein in its entirety.

All headings and subheadings are used herein for convenience only and are not to be construed as limiting the invention in any way.

The use of any and all embodiments or exemplary language (e.g., "such as") is intended to be illustrative only and not to limit the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating that any non-claimed elements are necessary to practice the invention.

The citation and incorporation of the patent documents herein is for convenience only and does not reflect any point of view of the validity, patentability and/or enforceability of such patent documents.

The present invention includes all modifications and equivalents of the subject matter described in the appended claims, as permitted by applicable law.

SEQ ID NO: 1

Lys-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Asn-Phe-Leu-Val-His-Ser-Ser-Asn-Asn-Phe-Gly-Ala- Ile-Leu-Ser-Ser-Thr-Asn-Val-Gly-Ser-Asn-Thr-Tyr

SEQ ID NO: 2

Lys-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Asn-Phe-Leu-Val-His-Sar-Ser-Asn-Asn-Phe-Gly-Pro- Ile-Leu-Pro-Pro-Thr-Asn-Val-Gly-Ser-Asn-Thr-Tyr

SEQ ID NO: 3

Xaa ₁ -Cys-Xaa ₃ -Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Glu-Phe-Leu-Xaa ₁₇ -Xaa ₁₈ -Ser-Ser-Xaa ₂₁ -Xaa ₂₂ - Phe-Gly-Pro-Xaa ₂₆ -Leu-Pro-Pro-Thr-Xaa ₃₁ -Val-Gly-Ser-Xaa ₃₅ -Thr-Xaa ₃₇

<110> Novo Nordisk A/S

<120> Amyloid derivatives

<130> 8383

<140> XXXXXX

<141> 2012-04-06

<160> 3

<170> PatentIn version 3.5

<210> 1

<211> 37

<212> PRT

<213> Human

<400> 1

<210> 2

<211> 37

<212> PRT

<213> Labor

<220>

<223> peptide

<400> 2

<210> 3

<211> 37

<212> PRT

<213> Labor

<220>

<223> peptide

<220>

<221> Miscellaneous Features

<222> (1)..(1)

<223> Xaa1 is deleted or independently selected from Ala, Cys, Glu, Gly, His, Arg, Sar, and Lys;

<220>

<221> Miscellaneous Features

<222> (3)..(3)

<223> Xaa3 is independently selected from the group consisting of Gly, His, Arg, Ser, and Asn;

<220>

<221> Miscellaneous Features

<222> (17)..(17)

<223> Xaa17 is independently selected from the group consisting of His, Arg, Lys and Val;

<220>

<221> Miscellaneous Features

<222> (18)..(18)

<223> Xaa18 is independently selected from Arg, Lys and His;

<220>

<221> Miscellaneous Features

<222> (21)..(21)

<223> Xaa21 is independently selected from the group consisting of Ala, Lys, Gln, Ser and Asn;

<220>

<221> Miscellaneous Features

<222> (22)..(22)

<223> Xaa22 is independently selected from the group consisting of Glu, Gln, Ser, Thr and Asn;

<220>

<221> Miscellaneous Features

<222> (26)..(26)

<223> Xaa26 is independently selected from Pro, Arg and Ile;

<220>

<221> Miscellaneous Features

<222> (31)..(31)

<223> Xaa31 is independently selected from Ser, Glu, Asp and Asn;

<220>

<221> Miscellaneous Features

<222> (35)..(35)

<223> Xaa35 is independently selected from the group consisting of His, Arg, Lys, Asp and Glu;

<220>

<221> Miscellaneous Features

<222> (37)..(37)

<223> Xaa37 is independently selected from Pro and Tyr;

<400> 3

Figure 1 presents a structure.

Figure 2 presents a structure.

Figure 3 presents a structure.

Figure 4 presents a structure.

Claims (15)

A multi-peptide comprising the amino acid sequence of the analog of SEQ ID No: 2, wherein: the analog comprises a glutamic acid residue at position 14; wherein the amino acid sequence number of the analog corresponds to SEQ ID No Amino acid numbering sequence of 2; and the restriction condition is that the multi-peptide is not N-α-(19-carboxynonanoyl)-Glu-Glu-Arg-[Glu1, Glu14, His17, Glu31]- Pramlintide; and the multi-peptide is optionally attached to at least one of its at least one amino acid residue. The multi-peptide of claim 1, wherein the analog comprises a residue at position 17, which is independently selected from any of histidine, arginine, lysine, and proline. Preferred; histidine and arginine; more preferably arginine. The multi-peptide of any one of claims 1 and 2, wherein the analog comprises a residue at position 37, the residue being independently selected from any one of valine and tyrosine; Preferred is valine acid. The multi-peptide of any one of claims 1 and 2, wherein the analog comprises a residue at position 35, the residue being independently selected from the group consisting of histidine, arginine, lysine, day Any of aspartic acid, glutamic acid, aspartic acid and glutamic acid; preferably histidine, aspartic acid and glutamic acid; more preferably histidine and aspartic acid Proline acid; more preferably aspartic acid. For example, the multi-peptide of any one of claims 1 and 2, wherein The analog comprises an amino acid residue at position 14 which is a glutamic acid residue, a residue at position 17 which is a arginine acid and a residue at position 37 which is a proline. The multi-peptide of claim 1, which comprises the amino acid sequence of the SEQ ID No: 2 analog of formula (I): Xaa ₁ -Cys-Xaa ₃ -Thr-Ala-Thr-Cys-Ala -Thr-Gln-Arg-Leu-Ala-Glu-Phe-Leu-Xaa ₁₇ -Xaa ₁₈ -Ser-Ser-Xaa ₂₁ -Xaa ₂₂ -Phe-Gly-Pro-Xaa ₂₆ -Leu-Pro-Pro-Thr- Xaa ₃₁ -Val-Gly-Ser-Xaa ₃₅ -Thr-Xaa ₃₇ ; Formula (I) (SEQ ID No: 3) wherein Xaa _{1 is} deleted or independently selected from Ala, Cys, Glu, Gly, His, Arg, Ser And Lys; Xaa _{3 is} independently selected from the group consisting of Gly, His, Arg, Ser and Asn; Xaa _{17 is} independently selected from the group consisting of His, Arg, Lys and Val; Xaa _{18 is} independently selected from Arg, Lys and His; Xaa _{21 is} independently selected From Ala, Lys, Gln, Ser and Asn; Xaa _{22 is} independently selected from Glu, Gln, Ser, Thr and Asn; Xaa _{26 is} independently selected from Pro, Arg and Ile; Xaa _{31 is} independently selected from Ser, Glu, Asp And Asn; Xaa _{35 is} independently selected from the group consisting of His, Arg, Lys, Asp, and Glu; Xaa _{37 is} independently selected from Pro and Tyr; and wherein the C-terminus can be derivatized as appropriate. The multi-peptide of any one of claims 1 and 2, wherein at least one substituent is attached to at least one amino acid residue of the multi-peptide. The multi-peptide of any one of claims 1 and 2, wherein the substituent is selected from any one of the following substituents: The multi-peptide of any one of claims 1 and 2, wherein the substituent is attached to the α-amino group of the N-terminal amino acid residue or to the Lys residue. The multi-peptide of any one of claims 1 and 2, which is selected from the group consisting of any of the following multi-peptides: N- α -[(S)-4-carboxy-4-( 19-carboxynonylnonylamino)butanyl]-Gly-[His1,Glu14,Arg17,Pro37]-pramlintide N- α -[(S)-4-carboxy-4-(19-carboxynonanthine ))]][His1,Glu14,Arg17,Pro37]-pramlintide N- α -[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Gly1,Glu14 ,Arg17,Pro37]-pramlintide N- α -[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Arg1,Glu14,Hi17,Pro37]-pramlintide N- α -[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Gly-[Arg1,Glu14,His17,Pro37]-pramlintide N- α -[(S )-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14, Arg17, Pro37]- pramlintide N- α -[(S)-4-carboxy-4-(19- Carboxyylamidosyl)butanyl]-[Glu14,His17,Gln21,Pro37]-pramlintide N- α -[(S)-4-carboxy-4-(17-carboxyheptylamino)butanyl] -[Glu14,His17,His35]-pramlintide N- α -[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14,His17,Pro37]-Planin Peptide N- α -[(S)-4-carboxylate 4-(19-carboxynonylamido)butanyl]-[Lys11,Glu14,His17,Pro37]-pramlintide N- α -[(S)-4-carboxy-4-(19-carboxy-ten Nonadenyl)butanyl]-Glu-Arg-[Glu1, Glu14, Arg17, Pro37]- pramlintide N- α -[(S)-4-carboxy-4-(19-carboxyundecylamino) Butyl]-[Glu14, Arg18, Ser21, Ser22, Ser28, Ser29, Asp31, Asp35, Pro37]- pramlintide N- α -[(S)-4-carboxy-4-(19-carboxynonylamino) ) 醯 醯]]-[Glu14, His17, Gln21, Gln35, Pro37]- pramlintide N- α -[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14, His17,Thr21,Pro37]-pramlintide N- α -[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[Glu14,His17,Gln35,Pro37]-Planin Peptide N- α -[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14, Arg17, His35]-planin Peptide N- α -[(S)-4 -carboxy-4-(19-carboxyundecylamino)butanyl]-Glu-[Glu14, Arg17, His35]- pramlintide N- α -[(S)-4-carboxy-4-(19-carboxyl 19-nonylamino)butanyl]-[Glu14,His17,Lys35]-pramlintide N- α- [2-(2-{2-[2-(2-{2-[(S)-4-carboxyl) -4-(19-carboxyheptadecylamido)butaninyl]ethoxy}ethoxy)acetamidine Amino]ethoxy}ethoxy)ethinyl]-D-Arg-D-Arg-[Glu14,His17,Arg18,Ala21,Ser35]-pramlintide N- α -[(S)-4- Carboxy-4-(19-carboxynonylamido)butanyl]-[Glu14,His17,Gln21,His35)-pramlintide N- ε- 21-[2-(2-{2-[2-(2) -{2-[(S)-4-carboxy-4-(17-carboxyheptadelamido)butanylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy Ethyl)-[Glu14,His17,Lys21,Glu35]-pramlintide N- α -[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-[His1, Glu14, His17, Arg18, Ser21, Ser22, Asp31, Asp35]- pramlintide N- α -[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-Arg- [Glu1, Glu14, Arg17, Gln21, Pro37]-Pullin peptide N- ε- 21-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-) (19-carboxyheptadelamido)butaninyl]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Arg18, Lys21, Ser28, Arg29,Arg35]-pramlintide N- α -[(S)-4-carboxy-4-(19-carboxynonylamido)butanyl]-Glu-Arg-[Glu1, Glu14, Arg17, Gln21, Gln35 ,Pro37]-pramlintide N- α -[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-Arg-[Glu1,Glu 14, Arg17, Gln21, Gln31, Gln35, Pro37]- pramlintide N- α -[(S)-4-carboxy-4-(19-carboxynonylamino)butanyl]-Glu-Arg-[Glu1 , Glu14, Arg17, Gln21, Gln22, Gln31, Gln35, Pro37]- pramlintide N- ε -17-[2-(2-{2-[2-(2-{2-[(S)-4-) Carboxy-4-(17-carboxyheptadecylamido)butanylamino]ethoxy}ethoxy)acetamido]ethoxy}ethoxy)ethinyl]-[Glu14, Lys17, Ser21, Glu35]-pramlintide. The multi-peptide of any one of claims 1 and 2, wherein the multi-peptide is: N-α-[(S)-4-carboxy-4-(19-carboxyundecylamino) Butyl]-[Glu14, Arg17, Pro37]-pramlintide [Example 53]. One of the first to eleventh patent applications for use as a pharmaceutical product One of the many peptides. A multi-peptide, as described in any one of claims 1 to 11, for the treatment or prevention of hyperglycemia, type 2 diabetes, abnormal glucose tolerance, type 1 diabetes, obesity, hypertension , Syndrome X, dyslipidemia, cognitive disorders, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, dyspepsia and gastric ulcers, and/or for reducing food intake, reducing beta- Cell apoptosis, increase of β-cell function and β-cell population, and/or restore glucose sensitivity of β-cells, hypercalcemia, osteoporosis and deformed osteitis. A pharmaceutical composition comprising the multipeptide of any one of claims 1 to 11 and a pharmaceutically acceptable excipient. A method of preparing a pharmaceutical composition according to claim 14, which comprises mixing the multipeptide of any one of claims 1 to 11 with at least one pharmaceutically acceptable excipient.