Polypeptides or analogs thereof and uses thereof

Abstract

The present application discloses a kind of polypeptide or its derivative or their pharmaceutically acceptable salt, it has the structure shown in formula (I) (U-X1SHX4SX6X7X8X9X 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 DX 22 PX 24 TDVGAGSX 32 -NH2) and modification group, the structure shown in formula (I) contains two amino acids Z1 or contains two amino acids Z2, two amino acids Z1 or two amino acids Z2 respectively X i And X i+7 , i is any integer between 6-13, modification group is connected with two amino acids Z1 or two amino acids Z2 by covalent bond form.This polypeptide or its derivative has certain activation activity to amylin receptor and calcitonin receptor simultaneously, and has longer in-vivo half-life by the modification of side chain modification group, to support low-dose, low-frequency administration, can be used for preventing or treating metabolic related diseases.

Description

Technical Field

[0001] This invention belongs to the field of biopharmaceutical technology, specifically relating to a polypeptide or its analogue and its application, and more specifically to a polypeptide or its derivative or pharmaceutically acceptable salts, pharmaceutical compositions and their uses that simultaneously activate amylin receptor (AMYR) and calcitonin receptor (CTR). Background Technology

[0002] The continued rise in the prevalence of obesity and type 2 diabetes mellitus (T2DM) has resulted in a lack of effective treatments. Furthermore, obese patients and those with T2DM often face a high risk of cardiovascular disease and other metabolic complications, complicating the safe use of medications. While bariatric surgery is currently effective in alleviating obesity and T2DM, it carries risks and involves irreversible bodily changes. Therefore, there is a need to develop novel and more effective pharmacological treatments for glycemic control and weight management.

[0003] Amylin is a pancreatic hormone composed of 37 amino acids, secreted by pancreatic β-cells along with insulin. It plays a role in satiety, gastric emptying, and glucagon secretion. Amylin controls satiety signals by directly acting on amylin receptors (AMYRs) in the brain. Three amylin receptor subtypes are currently known: Amy1R, Amy2R, and Amy3R. Studies have shown that Amy1R is associated with lipid metabolism, while Amy3R is responsible for blood glucose regulation. Amylin is also a potent inhibitor of gastric emptying and can further inhibit glucagon secretion through central mechanisms.

[0004] Calcitonin is a 32-amino acid peptide secreted by thyroid C cells. Calcitonin acts on the calcitonin receptor (CTR), lowering blood calcium levels by inhibiting osteoclasts and promoting renal calcium excretion.

[0005] Studies have shown that synergistic activation of amylin receptors and calcitonin receptors has a positive metabolic-promoting effect, effectively reducing blood glucose levels and weight. The development of long-acting dual agonists of amylin and calcitonin receptors holds promise for providing an effective drug for treating metabolic diseases, including obesity, type 2 diabetes, non-alcoholic fatty liver disease (NASH), and dyslipidemia. Currently, no long-acting dual agonists of amylin and calcitonin receptors (DACRAs) have been approved. Therefore, there is an urgent need to develop a peptide or analogue that simultaneously activates both amylin receptor (AMYR) and calcitonin receptor (CTR). Summary of the Invention

[0006] The present invention aims to at least partially address one of the technical problems existing in the prior art. To this end, the present invention provides a polypeptide or analogue thereof that simultaneously exhibits activating activity against both the amylin receptor (AMYR) and the calcitonin receptor (CTR).

[0007] This invention is based on the following discoveries of the inventors:

[0008] Several long-acting amylin analogs are in clinical trials and have shown certain advantages in treating metabolic diseases. Pramlintide is an amylin analog approved for use in combination with insulin therapy to improve insulin sensitivity and help lower blood sugar in diabetic patients. However, this drug has a half-life of less than one hour in vivo, requiring patients to inject it multiple times daily with meals, which is inconvenient. Furthermore, amylin analogs, due to their structural characteristics, are prone to fibrotic aggregation in vivo and in vitro, resulting in demanding storage conditions and potential immunogenic risks.

[0009] Salmon calcitonin (sCT) is used to treat hypercalcemia, osteoporosis, and osteitis deformans. Recent studies have shown that calcitonin also has other metabolic benefits, such as affecting insulin sensitivity, reducing gastric emptying rate, and promoting satiety. However, its short half-life in vivo necessitates multiple daily subcutaneous injections, leading to poor patient compliance. Furthermore, sCT has poor stability, is prone to aggregation and causing immunogenic reactions, raising safety concerns.

[0010] The development of peptide drugs is often hampered by short half-lives and low bioavailability, leading patients to receive higher doses more frequently, which may result in decreased adherence, increased costs, and increased risk of side effects. Therefore, it is necessary to develop therapeutic agents with extended half-lives.

[0011] Based on this, the method of the present invention introduces residues containing -SH or -NH2 side chains (including but not limited to cysteine ​​(C or c) or lysine (K or k)) by two-point mutation of the polypeptide chain, and performs two-site fixation modification of the side chain through macrocyclization reaction, thereby locking the preferred binding conformation of the target polypeptide. Compared with traditional polypeptide drugs, this strategy can reduce the tendency of aggregation and fibrosis, which is beneficial to further improve the stability of the target polypeptide and receptor binding activity.

[0012] Therefore, in one aspect of the invention, a polypeptide or its derivative, or a pharmaceutically acceptable salt thereof, is provided. According to embodiments of the invention, the polypeptide or its derivative, or a pharmaceutically acceptable salt thereof, has the structure shown in formula (I):

[0013] U-X1SHX4SX6X7X8X9X10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 DX 22 PX 24 TDVGAGSX 32 -NH2(I);

[0014] Each U is independently hydrogen, acetyl, or benzoyl;

[0015] X1 is A or K; X4 is L or αMeL; X6 is Z1, Z2, or T; X7 is Z1, Z2, or A; X8 is Z1, Z2, or V; X9 is Z1, Z2, or L; X 10 Z1, Z2, or G; X 11 Z1, Z2, homoR, Orn, or R; X 12 Z1, Z2, or L; X 13 Z1, Z2, or S; X 14 Z1, Z2, or A; X 15 Z1, Z2, or E; X 16 Z1, Z2, or L; X 17 Z1, Z2, or H; X 18 Z1, Z2, K, αMeK, or Orn; X 19 Z1, Z2, or L; X 20 Z1, Z2, or Aib; X 22 For Y or αMeF; X 24 For R, N-Me-R, homoR, norR, Q, or r; X 32 It can be P, Hyp, cis-P(4-NH2), trans-P(4-NH2), or p;

[0016] The structure shown in formula (I) contains two amino acids Z1 or two amino acids Z2, and the positions of the two amino acids Z1 or the two amino acids Z2 are X, respectively. i and X i+7 , where i is any integer between 6 and 13;

[0017] Each amino acid Z1 is independently selected from C, c, αMeC, HoC, Hoc, Pen, or N-Me-C;

[0018] Each amino acid Z2 is independently selected from K, k, Dap, Dab, Orn, HomoK, N-Me-K, N-Me-k, αMeK, and αMek.

[0019] The polypeptides or their derivatives, or pharmaceutically acceptable salts thereof (hereinafter referred to as polypeptides or analogues) according to embodiments of the present invention are dual-agonist polypeptide analogues that can simultaneously exhibit balanced or unbalanced activation activity against both the amylin receptor (AMYR) and the calcitonin receptor (CTR). They can be used for the prevention or treatment of metabolic disorders such as type 2 diabetes mellitus (T2DM), obesity, hyperlipidemia, NAFLD, and NASH; they also have potential applications in the treatment of other diseases, such as symptoms or diseases related to Alzheimer's disease (AD), alcohol or drug addiction, etc.

[0020] In a second aspect, the present invention provides a pharmaceutical composition. According to embodiments of the invention, the pharmaceutical composition comprises the polypeptide or its derivatives or pharmaceutically acceptable salts described in the first aspect. As is previously known, polypeptides or their derivatives or pharmaceutically acceptable salts (hereinafter referred to as polypeptides or analogs) are dual-agonist polypeptide analogs, which can simultaneously possess a certain degree of balanced or unbalanced activation activity against both the amylin receptor (AMYR) and the calcitonin receptor (CTR). Therefore, the use of a drug containing polypeptides or their derivatives or pharmaceutically acceptable salts can effectively prevent or treat diseases related to the amylin receptor and / or calcitonin receptor.

[0021] In a third aspect, the present invention provides a combination drug or kit. According to embodiments of the invention, the combination drug or kit comprises: a polypeptide or derivative thereof described in the first aspect, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described in the second aspect as a first active ingredient; and a second active ingredient; wherein the second active ingredient comprises a drug for the prevention and / or treatment of a disease. The combination drug or kit according to embodiments of the invention can further improve the therapeutic effect on diseases related to amylin receptors and / or calcitonin receptors.

[0022] In a fourth aspect of the invention, the invention provides for the use of the polypeptides or derivatives thereof described in the first aspect, or pharmaceutically acceptable salts thereof, the pharmaceutical compositions described in the second aspect, or the combination drugs or cassettes described in the third aspect, in the preparation of a medicament for the treatment or prevention of diseases related to amylin receptors and / or calcitonin receptors.

[0023] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Attached Figure Description

[0024] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which:

[0025] Figure 1 This is a schematic diagram of the Staple linker synthesis process in Embodiment 1 of the present invention;

[0026] Figure 2 This is a graph showing the binding activity results of different polypeptide derivatives with AMY3R in Test Example 1 of this invention;

[0027] Figure 3 This is a graph showing the binding activity results of different polypeptide derivatives with CTR in Test Example 1 of the present invention;

[0028] Figure 4 shows the p-pharmaceutical curves of different polypeptide derivatives in rats in Test Example 2 of the present invention.

[0029] Figure 5 This is a p-pharmaceutical curve of different polypeptide derivatives in cynomolgus monkeys in Test Example 2 of the present invention;

[0030] Figure 6 The results of changes in body weight and food intake after a single administration of the polypeptide derivative in SD rats in Test Example 3 of this invention are shown.

[0031] Figure 7 The results of changes in body weight and food intake after a single administration of the polypeptide derivative in the DIO mouse model in Test Example 4 of this invention are shown. The positive control drug is canglitazone.

[0032] Figure 8 The results of a single administration of the polypeptide derivative combined with semaglutide in the DIO mouse model in Test Example 5 of this invention showed the changes in body weight and food intake. The positive control drugs were the same dose of canglitide or the same dose of semaglutide.

[0033] Figure 9 The results of continuous administration of the polypeptide derivative in the ZDF rat model in Test Example 6 of this invention showed changes in body weight, blood glucose, and food intake.

[0034] Figure 10 The results of the polypeptide aggregation and precipitation stability test in Test Example 9 of this invention are shown. The positive control is salmon calcitonin. Detailed Implementation

[0035] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0036] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. Furthermore, in the description of this invention, unless otherwise stated, "a plurality of" means two or more.

[0037] Detailed description of the invention

[0038] Definitions and General Terms

[0039] In this document, the terms “comprising” or “including” are open-ended expressions, meaning that they include the contents specified in this invention, but do not exclude other aspects.

[0040] In this document, the term "pharmaceutical acceptable" means that a substance or composition must be chemically and / or toxicologically compatible with other components comprising a polypeptide or its derivatives and / or with the mammals to which it is treated. Preferably, "pharmaceutical acceptable" as used herein means approved by a federal regulatory agency or national government, or listed in the United States Pharmacopeia or other generally recognized pharmacopoeia for use in animals, particularly in humans.

[0041] In this document, the term "pharmaceutically acceptable salt" refers to the organic and inorganic salts of the polypeptides or their derivatives of the present invention. Pharmaceutically acceptable salts are well known in the field, as described in the literature: SMBerge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66: 1-19.

[0042] In this document, amino acids are represented by the conventional single-letter and three-letter codes for natural amino acids, as well as the generally accepted three-letter codes for other α-amino acids; for example, α-aminoisobutyric acid can be represented by either Aib or B. Unless otherwise specified, all uppercase amino acid residues in this invention are in the L-configuration, and lowercase amino acid residues are in the D-configuration.

[0043] In this article, the term "Aib" is structured as follows:

[0044] In this paper, the structural formula for the term "αMeF" is:

[0045] In this article, the structural formula for the term "Orn" is:

[0046] In this paper, the structural formula for the term "αMeL" is:

[0047] In this paper, the structural formula for the term "αMeK" is:

[0048] In this article, the structural formula for the term "N-Me-R" is:

[0049] In this article, the structural formula for the term "Hyp" is:

[0050] In this article, the structural formula for the term "homoR" is:

[0051] In this paper, the structural formula of the term "cis-P(4-NH2)" is:

[0052] In this paper, the structural formula of the term "trans-P(4-NH2)" is: In this paper, the structural formula for the term "αMeC" is: In this article, the structural formula for the term "HoC" is: In this article, the structural formula for the term "Hoc" is: In this article, the structural formula for the term "Pen" is: In this article, the structural formula for the term "N-Me-C" is: In this article, the structural formula for the term "Dab" is: In this article, the structural formula for the term "Dap" is: In this article, the structural formula for the term "homoK" is: In this paper, the structural formula for the term "αMek" is:

[0053] In this article, the structural formula for the term "N-Me-K" is:

[0054] In this document, the terms “optionally,” “optionally,” or “optionally” generally refer to an event or condition that may, but may not, occur, and the description includes both cases in which the event or condition occurs and cases in which the event or condition does not occur.

[0055] In this document, the terms "optionally substituted," "optionally substituted," and "substituted or unsubstituted" are used interchangeably. Generally, the term "optionally," whether preceding or following the term "substituted," indicates that one or more hydrogen atoms in the given structure are substituted by a specific substituent. Unless otherwise indicated, an optional substituent may be substituted at each substituted position of the group. When more than one position in the given structural formula can be substituted by one or more substituents selected from a specific group, the substituents may be substituted at the same or different positions. The substituents mentioned may be, but are not limited to, F, Cl, Br, CN, OH, NH2, NO2, etc.

[0056] In this document, the term "one or more" (e.g., in the definition of substituents in compounds (modifying groups) of the general formula of the present invention) means "one, two, three, four or five, especially one, two, three or four, more especially one, two or three, and even more especially one or two".

[0057] Additionally, it should be noted that, unless otherwise explicitly stated, the descriptive terms “each…independently is”, “…each independently is”, and “…independently is” used in this invention are interchangeable and should be interpreted broadly. They can mean that the specific options expressed by the same symbols in different groups do not affect each other, or that the specific options expressed by the same symbols in the same group do not affect each other.

[0058] In this article, the term "halogen" refers to a fluorine, chlorine, bromine, or iodine atom.

[0059] In this paper, the minimum and maximum carbon atom content in hydrocarbon groups are indicated by prefixes, for example, prefix C. a～b This refers to a carbon atom, which is "a" to "b". For example, "C 1～n "C" refers to a saturated / unsaturated carbon chain, either straight or branched, containing 1, 2, 3, 4, 5, ..., or n carbon atoms; further understanding, "C" 1～n "Should be interpreted as any subranges included, such as C" 1～20 In, containing C 1～20 C 1～18 C 10～8 C 12～18 C 14～18 C 1～10 C 1～6 C 1～3 C 1～2 C 2～10 C 2～9 C 2～8 C 2～7 C 2～6 C2～5 C 2～4 C 2～3 C 3～10 C 3～9 C 3～8 C 3～7 C 3～6 C 3～5 C 3～4 C 4～10 C 4～9 C 4～8 C 4～7 C 4～6 C 4～5 .

[0060] In this article, the general structural formula of the term "alkyl" is: Alkylenes can be straight-chain alkyl or branched-chain alkyl. For example, the term "C"... 1～20 "Alkyl" refers to an alkyl group having 1 to 20 carbon atoms; the term "C" 1～6 "Alkyl" refers to an alkyl group having 1 to 6 carbon atoms.

[0061] In this article, the general structural formula of the term "alkylene" is: Alkylenes can be straight-chain or branched. For example, the term "C"... 1～20 "alkylene" refers to an alkylene group having 1 to 20 carbon atoms; the term "C" 1～6 "Alkylene" refers to an alkylene group having 1 to 6 carbon atoms.

[0062] In this article, the term "C" 1～6 "Alkoxy" refers to a C-type compound containing the formula "-O-alkyl". 1～6 Alkyl, wherein the term "alkyl" is as defined above. Examples include: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, sec-butoxy, pentoxy, isopentoxy, and n-hexyloxy, or isomers of the above groups. In particular, the "C" group... 1～6 An alkoxy group can contain 1, 2, 3, 4, or 5 carbon atoms ("C"). 1～5 Alkoxy group), preferably, may contain 1, 2, 3 or 4 carbon atoms ("C"). 1～4 (alkoxy group).

[0063] In this article, the term "oxyalkylene" refers to the group formed by removing one hydrogen atom from an "oxyalkylene" group.

[0064] In this document, the term "aryl" refers to a carbocyclic system containing monocyclic, bicyclic, and tricyclic rings, wherein at least one ring system is aromatic, and each ring system comprises a ring of 3-10 atoms. The aryl group is typically, but not necessarily, linked to the parent molecule via an aromatic ring. The term "aryl" may be used interchangeably with the terms "aromatic ring" or "aromatic ring". Aryl groups may include phenyl, indenyl, naphthyl, and anthraceneyl groups. The aryl group may optionally be substituted by one or more substituents described in this disclosure.

[0065] In this article, the term "aryl" refers to the group formed by removing one more hydrogen atom from an "aryl" group.

[0066] In this paper, the term "heteroaryl" refers to a carbocyclic system containing monocyclic, bicyclic, and tricyclic rings, wherein at least one ring system is aromatic. Heteroatoms include nitrogen, oxygen, sulfur, etc. It typically represents a monovalent saturated or partially unsaturated monocyclic or bicyclic ring system with multiple ring atoms, containing one, two, or three cyclic heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon.

[0067] In this paper, the term "hybrid aryl" refers to the group formed by removing one more hydrogen atom from a "heteroaryl" group.

[0068] In the description of the functional groups of this invention It is used to describe the position of the substituent group.

[0069] In this paper, the structural formula of the term "-NH-CO-" or "-NH-C(=O)-" is:

[0070] In this document, "pharmaceutical composition" can refer to a drug for the treatment of a disease or for use in in vitro cell culture experiments. When used for the treatment of a disease, the term "pharmaceutical composition" generally refers to a unit dose form, which can be of any type of formulation and can be prepared by any method well known in the pharmaceutical industry. All methods involve the step of combining the active ingredient with excipients constituting one or more adjunct components. Typically, compositions are prepared by uniformly and adequately combining an active polypeptide or its derivative or revitalizer with a liquid excipient, a finely ground solid excipient, or both.

[0071] In the chemical structure of the ligands or compounds described in this disclosure, the bonds... This indicates that the configuration is not specified. If chiral isomers exist in the chemical structure, the bond... It can be Or simultaneously include Two configurations. Although all the above structural formulas are shown in some isomer forms for simplicity, this disclosure can include all isomers, such as tautomers, rotational isomers, geometric isomers, diastereomers, racemates, and enantiomers.

[0072] In this document, the term "pharmaceuticalally acceptable excipient" may include any solvent, including but not limited to pharmaceutically acceptable carriers, stabilizers, dispersants, cosolvents, plasticizers, solid excipients, diluents, or other liquid excipients, etc., suitable for a particular target dosage form. The use of any conventional excipients is also within the scope of this invention, except for any range of incompatibilities with the polypeptides or derivatives thereof, pharmaceutical compositions, or drugs containing them, such as any adverse biological effects or harmful interactions with any other component of a pharmaceutically acceptable composition.

[0073] In addition to any conventional excipients, the use of polypeptides or their derivatives, pharmaceutical compositions or pharmaceuticals containing them that are incompatible with the present invention, such as any adverse biological effects or harmful interactions with any other component of a pharmaceutically acceptable composition, is also within the scope of this invention.

[0074] The pharmaceutical compositions disclosed herein include formulations suitable for parenteral administration. The formulations can be conveniently available in unit dosage forms and can be prepared by any method known in the pharmaceutical field. The amount of active ingredient in a single-dose form, which can be prepared in combination with excipients, is generally the amount of the polypeptide or a derivative thereof that produces the therapeutic effect.

[0075] In this paper, the term "agonist" refers to a substance (ligand) that activates the type of receptor.

[0076] In this document, the term "treatment" refers to the administration of a drug to achieve a desired pharmacological and / or physiological effect. This effect may be preventative in terms of complete or partial prevention of a disease or its symptoms, and / or therapeutic in terms of partial or complete cure of a disease and / or adverse effects caused by the disease. As used herein, "treatment" encompasses diseases in mammals, particularly humans, including: (a) prevention of disease or the onset of a condition in individuals susceptible to disease but not yet diagnosed with the disease; (b) inhibition of disease, such as blocking disease progression; or (c) relief of disease, such as reducing disease-related symptoms. As used herein, "treatment" encompasses any administration of a drug containing a peptide or its derivative to an individual to treat, cure, relieve, improve, reduce, or inhibit the individual's disease, including but not limited to administration of a drug containing a peptide or its derivative described herein to an individual in need.

[0077] In this article, the term “non-alcoholic fatty liver disease (NAFLD)” generally refers to a clinicopathological syndrome characterized by excessive fat deposition in hepatocytes excluding alcohol and other clearly defined liver-damaging factors. It is an acquired metabolic stress-induced liver injury closely associated with insulin resistance and genetic susceptibility, including but not limited to simple fatty liver (SFL), non-alcoholic steatohepatitis (NASH), and its associated cirrhosis.

[0078] Detailed description of the polypeptides or analogues of the present invention and their applications

[0079] This invention proposes a polypeptide or its derivative, or their pharmaceutically acceptable salts, pharmaceutical compositions, and their uses, which will be described in detail below.

[0080] polypeptides or their derivatives or their pharmaceutically acceptable salts

[0081] In one aspect of the invention, a polypeptide or a derivative thereof, or a pharmaceutically acceptable salt thereof, is provided. According to embodiments of the invention, the polypeptide or a derivative thereof, or a pharmaceutically acceptable salt thereof, has the structure shown in formula (I):

[0082] U-X1SHX4SX6X7X8X9X 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 X 18 X 19 X 20 DX 22 PX 24 TDVGAGSX 32 -NH2(I);

[0083] Each U is independently hydrogen, acetyl, or benzoyl;

[0084] X1 is A or K; X4 is L or αMeL; X6 is Z1, Z2, or T; X7 is Z1, Z2, or A; X8 is Z1, Z2, or V; X9 is Z1, Z2, or L; X 10 Z1, Z2, or G; X 11 Z1, Z2, homoR, Orn, or R; X 12 Z1, Z2, or L; X 13 Z1, Z2, or S; X 14 Z1, Z2, or A; X 15 Z1, Z2, or E; X 16 Z1, Z2, or L; X 17 Z1, Z2, or H; X18 Z1, Z2, K, αMeK, or Orn; X 19 Z1, Z2, or L; X 20 Z1, Z2, or Aib; X 22 For Y or αMeF; X 24 For R, N-Me-R, homoR, norR, Q, or r; X 32 It can be P, Hyp, cis-P(4-NH2), trans-P(4-NH2), or p;

[0085] The structure shown in formula (I) contains two amino acids Z1 or two amino acids Z2, and the positions of the two amino acids Z1 or the two amino acids Z2 are X, respectively. i and X i+7 , where i is any integer between 6 and 13;

[0086] Each amino acid Z1 is independently selected from C, c, αMeC, HoC, Hoc, Pen, or N-Me-C;

[0087] Each amino acid Z2 is independently selected from K, k, Dap, Dab, Orn, HomoK, N-Me-K, N-Me-k, αMeK, and αMek.

[0088] The polypeptides or their derivatives, or pharmaceutically acceptable salts thereof (hereinafter referred to as polypeptides or analogues) according to embodiments of the present invention are dual-agonist polypeptide analogues that can simultaneously exhibit balanced or unbalanced activation activity against both the amylin receptor (AMYR) and the calcitonin receptor (CTR). They can be used for the prevention or treatment of metabolic disorders such as type 2 diabetes mellitus (T2DM), obesity, hyperlipidemia, NAFLD, and NASH; they also have potential applications in the treatment of other diseases, such as symptoms or diseases related to Alzheimer's disease (AD), alcohol or drug addiction, etc.

[0089] Furthermore, the peptides or their analogues described above can be modified by altering two amino acids Z1 or two amino acids Z2 to achieve a longer in vivo half-life, thus supporting low-dose, low-frequency administration.

[0090] It should be noted that, unless otherwise specified, U in this article refers to a hydrogen atom or a substituent group in the -NH2 group of amino acid X1. When U is hydrogen, the -NH2 group of X1 is -NH2 itself; when U is acetyl (i.e., Ac), the -NH2 group of X1 is -NHAc; when U is benzoyl (i.e., Bzl), the -NH2 group of X1 is -NH(Bzl).

[0091] It should be noted that, unless otherwise specified in this article, X 32 -NH2 refers to X 32The -OH in -COOH is replaced by -NH2, i.e., -CONH2.

[0092] According to an embodiment of the present invention, the positions of the two amino acids Z1 or the two amino acids Z2 are one of the following groups: X6 and X 13 X7 and X 14 X8 and X 15 X9 and X 16 X 10 and X 17 X 11 and X 18 X 12 and X 19 X 13 and X 20 .

[0093] According to an embodiment of the present invention, the positions of the two amino acids Z1 or the two amino acids Z2 are X8 and X, respectively. 15 .

[0094] According to embodiments of the present invention, the two amino acids Z1 or the two amino acids Z2 are each independently selected from K, C or C.

[0095] According to an embodiment of the present invention, X1 is A or K.

[0096] According to an embodiment of the present invention, X1 is A.

[0097] According to an embodiment of the present invention, X4 is L.

[0098] According to an embodiment of the present invention, X6 is K, c, C or T.

[0099] According to an embodiment of the present invention, X6 is C or T.

[0100] According to an embodiment of the present invention, X6 is T.

[0101] According to an embodiment of the present invention, X7 is K, c, C or A.

[0102] According to an embodiment of the present invention, X7 is C or A.

[0103] According to an embodiment of the present invention, X7 is A.

[0104] According to an embodiment of the present invention, X8 is C, V, c, HoC, Pen, αMeC, N-Me-C, K, Orn, k, or Dab.

[0105] According to an embodiment of the present invention, X8 is K, c, C or V.

[0106] According to an embodiment of the present invention, X8 is C, V or K.

[0107] According to an embodiment of the present invention, X8 is C or V.

[0108] According to an embodiment of the present invention, X8 is V.

[0109] According to an embodiment of the present invention, X9 is K, c, C or L.

[0110] According to an embodiment of the present invention, X9 is C or L.

[0111] According to an embodiment of the present invention, X9 is L.

[0112] According to an embodiment of the present invention, X 10 It can be K, c, C, or G.

[0113] According to an embodiment of the present invention, X 10 It is either C or G.

[0114] According to an embodiment of the present invention, X 10 It is G.

[0115] According to an embodiment of the present invention, X 11 For K, c, C, homoR, Orn, or R.

[0116] According to an embodiment of the present invention, X 11 It can be K, C, homoR, Orn, or R.

[0117] According to an embodiment of the present invention, X 11 For K, C, homoR, or R.

[0118] According to an embodiment of the present invention, X 11 For C, homoR, or R.

[0119] According to an embodiment of the present invention, X 11 For homoR or R.

[0120] According to an embodiment of the present invention, X 12 It can be K, c, C, or L.

[0121] According to an embodiment of the present invention, X 12 It can be C or L.

[0122] According to an embodiment of the present invention, X 12 Let L be the value.

[0123] According to an embodiment of the present invention, X 13 It can be K, c, C, or S.

[0124] According to an embodiment of the present invention, X 13 It can be C or S.

[0125] According to an embodiment of the present invention, X 13 Let it be S.

[0126] According to an embodiment of the present invention, X 14 It can be K, c, C, or A.

[0127] According to an embodiment of the present invention, X 14 It can be either C or A.

[0128] According to an embodiment of the present invention, X 14 The answer is A.

[0129] According to an embodiment of the present invention, X 15 For C, E, c, HoC, Pen, αMeC, αMeK, N-Me-C, K, Orn, k, or Dab.

[0130] According to an embodiment of the present invention, X 15 It can be K, c, C, or E.

[0131] According to an embodiment of the present invention, X 15 It can be K, C, or E.

[0132] According to an embodiment of the present invention, X 15 It can be C or E.

[0133] According to an embodiment of the present invention, X 15 The answer is C.

[0134] According to an embodiment of the present invention, X 16 It can be K, c, C, or L.

[0135] According to an embodiment of the present invention, X 16 It can be C or L.

[0136] According to an embodiment of the present invention, X 16 Let L be the value.

[0137] According to an embodiment of the present invention, X 17 It can be K, c, C, or H.

[0138] According to an embodiment of the present invention, X 17 It is C or H.

[0139] According to an embodiment of the present invention, X 17 For H.

[0140] According to an embodiment of the present invention, X 18 For c, K, C, k, αMeK, or Orn.

[0141] According to an embodiment of the present invention, X 18 For K, C, k, αMeK, or Orn.

[0142] According to an embodiment of the present invention, X 18 It can be C, K, or αMeK.

[0143] According to an embodiment of the present invention, X 18 It is K or αMeK.

[0144] According to an embodiment of the present invention, X 19 It can be K, c, C, or L.

[0145] According to an embodiment of the present invention, X 19 It can be C or L.

[0146] According to an embodiment of the present invention, X 21 Let L be the value.

[0147] According to an embodiment of the present invention, X 20 It can be K, c, C, or Aib.

[0148] According to an embodiment of the present invention, X 20 It is either C or Aib.

[0149] According to an embodiment of the present invention, X 20 It is Aib.

[0150] According to an embodiment of the present invention, X 22 The value is Y.

[0151] According to an embodiment of the present invention, X 23 Let P be the value.

[0152] According to an embodiment of the present invention, X 24 For R, N-Me-R, homoR, norR, r, or Q.

[0153] According to an embodiment of the present invention, X 24 It can be R, N-Me-R, homoR, norR, or Q.

[0154] According to an embodiment of the present invention, X 24 For R, N-Me-R, or Q.

[0155] According to an embodiment of the present invention, X 29 The answer is A.

[0156] According to an embodiment of the present invention, X 32 For P or Hyp.

[0157] According to an embodiment of the present invention, the structure shown in formula (I) has the following structure: U-X1SHX4STX7X8LGX 11 LX 13 X 14 X 15 LHX18 LX 20 DX 22 X 23 X 24 TDVGAGSX 32 -NH2(Ia).

[0158] In some alternative embodiments of the present invention, in the structure shown in formula (Ia), X1 is A or K.

[0159] In some alternative embodiments of the present invention, in the structure shown in formula (Ia), X4 is L or αMeL.

[0160] In some alternative embodiments of the present invention, in the structure shown in formula (Ia), X7 is C or A.

[0161] In some alternative embodiments of the present invention, in the structure shown in formula (Ia), X8 is C or V.

[0162] In some alternative embodiments of the present invention, in the structure shown in formula (Ia), X 11 For homoR, C, or R.

[0163] In some alternative embodiments of the present invention, in the structure shown in formula (Ia), X 13 It can be C or S.

[0164] In some alternative embodiments of the present invention, in the structure shown in formula (Ia), X 14 It can be either C or A.

[0165] In some alternative embodiments of the present invention, in the structure shown in formula (Ia), X 15 It can be C or E.

[0166] In some alternative embodiments of the present invention, in the structure shown in formula (Ia), X 18 For C, K, k, αMeK, or Orn.

[0167] In some alternative embodiments of the present invention, in the structure shown in formula (Ia), X 20 It is either C or Aib.

[0168] In some alternative embodiments of the present invention, in the structure shown in formula (Ia), X 22 It is either Y or αMeF.

[0169] In some alternative embodiments of the present invention, in the structure shown in formula (Ia), X 23 It can be P or K.

[0170] In some alternative embodiments of the present invention, in the structure shown in formula (Ia), X24 It can be R, N-Me-R, homoR, norR, or Q.

[0171] In some alternative embodiments of the present invention, in the structure shown in formula (Ia), X 32 For P or Hyp.

[0172] According to an embodiment of the present invention, the structure shown in formula (I) has the following structure: U-X1SHX4STAX8LGX 11 LSAX 15 LHX 18 L-Aib-DX 22 PX 24 TDVGAGSX 32 -NH2(Ib).

[0173] In some alternative embodiments of the present invention, in the structure shown in formula (Ib), X1 is A or K.

[0174] In some alternative embodiments of the present invention, in the structure shown in formula (Ib), X4 is L or αMeL.

[0175] In some alternative embodiments of the present invention, in the structure shown in formula (Ib), X8 is C, V, c, HoC, Pen, αMeC, N-Me-C, K, Orn, k, or Dab.

[0176] In some alternative embodiments of the present invention, in the structure shown in formula (Ib), X 11 For homoR, C, K, Orn, or R.

[0177] In some alternative embodiments of the present invention, in the structure shown in formula (Ib), X 15 For C, E, c, HoC, Pen, αMeC, αMeK, N-Me-C, K, Orn, k, or Dab.

[0178] In some alternative embodiments of the present invention, in the structure shown in formula (Ib), X 18 It can be C, K, αMeK, or Orn.

[0179] In some alternative embodiments of the present invention, in the structure shown in formula (Ib), X 22 It is either Y or αMeF.

[0180] In some alternative embodiments of the present invention, in the structure shown in formula (Ib), X 24 For R, N-Me-R, homoR, norR, r, or Q.

[0181] In some alternative embodiments of the present invention, in the structure shown in formula (Ib), X 32 For P or Hyp.

[0182] According to an embodiment of the present invention, the structure shown in formula (I) has the following structure: U-ASHX4STX7X8LGX11X 12 X 13 X 14 X 15 LX 17 X 18 X 19 X 20 DX 22 PX 24 TDVGAGSX 32 -NH2(II).

[0183] In some alternative embodiments of the present invention, in the structure shown in formula (II), X4 is L or αMeL.

[0184] In some alternative embodiments of the present invention, in the structure shown in formula (II), X7 is C or A.

[0185] In some alternative embodiments of the present invention, in the structure shown in formula (II), X8 is C, K or V.

[0186] In some alternative embodiments of the present invention, in the structure shown in formula (II), X 11 For homoR, C, K or R.

[0187] In some alternative embodiments of the present invention, in the structure shown in formula (II), X 12 It can be L or C.

[0188] In some alternative embodiments of the present invention, in the structure shown in formula (II), X 13 It can be C or S.

[0189] In some alternative embodiments of the present invention, in the structure shown in formula (II), X 14 It can be either C or A.

[0190] In some alternative embodiments of the present invention, in the structure shown in formula (II), X 15 It can be K, C, or E.

[0191] In some alternative embodiments of the present invention, in the structure shown in formula (II), X 17 It is C or H.

[0192] In some alternative embodiments of the present invention, in the structure shown in formula (II), X 18 It can be C, K, αMeK, or Orn.

[0193] In some alternative embodiments of the present invention, in the structure shown in formula (II), X 19 It can be C or L.

[0194] In some alternative embodiments of the present invention, in the structure shown in formula (II), X 20 It is either C or Aib.

[0195] In some alternative embodiments of the present invention, in the structure shown in formula (II), X 22 It is either Y or αMeF.

[0196] In some alternative embodiments of the present invention, in the structure shown in formula (II), X 24 For R, N-Me-R, or Q.

[0197] In some alternative embodiments of the present invention, in the structure shown in formula (II), X 32 For P or Hyp.

[0198] According to an embodiment of the present invention, the structure shown in formula (I) has the following structure: U-ASHX4STAX8LGX 11 LX 13 AX 15 LHX 18 LX 20 DX 22 PX 24 TDVGAGSX 32 -NH2(IIa).

[0199] In some alternative embodiments of the present invention, in the structure shown in formula (IIa), X4 is L or αMeL.

[0200] In some alternative embodiments of the present invention, in the structure shown in formula (IIa), X8 is C, K or V.

[0201] In some alternative embodiments of the invention, in the structure shown in formula (IIa), X 11 For homoR, C, K or R.

[0202] In some alternative embodiments of the invention, in the structure shown in formula (IIa), X 13 It can be C or S.

[0203] In some alternative embodiments of the invention, in the structure shown in formula (IIa), X 15 It can be K, C, or E.

[0204] In some alternative embodiments of the invention, in the structure shown in formula (IIa), X 18It can be C, K, αMeK, or Orn.

[0205] In some alternative embodiments of the invention, in the structure shown in formula (IIa), X 20 It is either C or Aib.

[0206] In some alternative embodiments of the invention, in the structure shown in formula (IIa), X 22 It is either Y or αMeF.

[0207] In some alternative embodiments of the invention, in the structure shown in formula (IIa), X 24 For R, N-Me-R, or Q.

[0208] In some alternative embodiments of the invention, in the structure shown in formula (IIa), X 32 For P or Hyp.

[0209] According to an embodiment of the present invention, the structure shown in formula (I) has the following structure: U-ASHLSTAX8LGX 11 LSAX 15 LHX 18 L-Aib-DYPX 24 TDVGAGSX 32 -NH2(IIb).

[0210] In some alternative embodiments of the present invention, in the structure shown in formula (IIb), X8 is C, K or V.

[0211] In some alternative embodiments of the invention, in the structure shown in formula (IIb), X 11 For homoR, C, or R.

[0212] In some alternative embodiments of the invention, in the structure shown in formula (IIb), X 15 It can be K, C, or E.

[0213] In some alternative embodiments of the invention, in the structure shown in formula (IIb), X 18 It can be C, K, or αMeK.

[0214] In some alternative embodiments of the invention, in the structure shown in formula (IIb), X 24 For R, N-Me-R, or Q.

[0215] In some alternative embodiments of the invention, in the structure shown in formula (IIb), X 32 For P or Hyp.

[0216] According to an embodiment of the present invention, the structure shown in formula (I) has the following structure: U-ASHLSTACLGX 11 LSACLHX 18 L-Aib-DYPX 24 TDVGAGSX 32 -NH2(III).

[0217] In some alternative embodiments of the present invention, in the structure shown in formula (III), X 11 For homoR or R.

[0218] In some alternative embodiments of the present invention, in the structure shown in formula (III), X 18 It is K or αMeK.

[0219] In some alternative embodiments of the present invention, in the structure shown in formula (III), X 24 For R, N-Me-R, or Q.

[0220] In some alternative embodiments of the present invention, in the structure shown in formula (III), X 32 For P or Hyp.

[0221] According to an embodiment of the present invention, the structure shown in formula (I) has at least one of the following structures:

[0222] U-ASHLSCAVLGRLCAELHKL-Aib-DYPRTDVGAGSP-NH2;

[0223] U-ASHLSTCVLGRLSCELHKL-Aib-DYPRTDVGAGSP-NH2;

[0224] U-ASHLSTACLGRLSACLHKL-Aib-DYPRTDVGAGSP-NH2;

[0225] U-ASHLSTAVCGRLSAECHKL-Aib-DYPRTDVGAGSP-NH2;

[0226] U-ASHLSTAVLCRLSAELCKL-Aib-DYPRTDVGAGSP-NH2;

[0227] U-ASHLSTAVLGCLSAELHCL-Aib-DYPRTDVGAGSP-NH2;

[0228] U-ASHLSTAVLGRCSAELHKC-Aib-DYPRTDVGAGSP-NH2;

[0229] U-ASHLSTAVLGRLCAELHKLCDYPRTDVGAGSP-NH2;

[0230] U-ASHLSTACLGRLSACLH-Orn-L-Aib-D-(αMeF)-PRTDVGAGS-Hyp-NH2;

[0231] U-ASH-(αMeL)-STACLGRLSACLHKL-Aib-D-(αMeF)-P-(N-Me-R)-TDVGAGSP-NH2;

[0232] U-ASHLSTACLGRLSACLH-(αMeK)-L-Aib-DYP-(N-Me-R)-TDVGAGS-Hyp-NH2;

[0233] U-ASHLSTACLG-homoR-LSACLH-(αMeK)-L-Aib-DYP-(N-Me-R)-TDVGAGSP-NH2;

[0234] U-ASHLSTACLGRLSACLH-(αMeK)-L-Aib-DYP-homoR-TDVGAGS-Hyp-NH2;

[0235] U-ASHLSTACLGRLSACLH-(αMeK)-L-Aib-DYP-norR-TDVGAGS-Hyp-NH2;

[0236] U-ASHLSTACLGRLSACLH-(αMeK)-L-Aib-DYPQTDVGAGS-Hyp-NH2;

[0237] U-ASHLSTACLGRLSACLH-(αMeK)-L-Aib-DYP-r-TDVGAGS-Hyp-NH2;

[0238] U-ASHLSTAVLGCLSAELHCL-Aib-DYPQTDVGAGS-Hyp-NH2;

[0239] U-KSHLSTACLGRLSACLH-(αMeK)-L-Aib-DYP-(N-Me-R)-TDVGAGS-Hyp-NH2;

[0240] U-KSHLSTAVLGCLSAELHCL-Aib-DYPQTDVGAGS-Hyp-NH2;

[0241] U-KSHLSTA-c-LGRLSA-c-LH-(αMeK)-L-Aib-DYPQTDVGAGS-Hyp-NH2;

[0242] U-ASHLSTACLGRLSACLH-(αMeK)-L-Aib-DYPQTDVGAGS-(cis-P(4-NH2))-NH2;

[0243] U-ASHLSTACLGRLSACLH-(αMeK)-L-Aib-DYPQTDVGAGS-(trans-P(4-NH2))-NH2;

[0244] U-ASHLSTACLGRLSACLH-(αMeK)-L-Aib-DYPQTDVGAGS-p-NH2;

[0245] U-ASHLSTACLG-homoR-LSACLH-(αMeK)-L-Aib-DYPQTDVGAGS-Hyp-NH2;

[0246] U-ASHLSTACLG-Orn-LSA CLH-(αMeK)-L-Aib-DYPQTDVGAGS-Hyp-NH2;

[0247] U-ASHLSTA-HoC-LGRLSA-HoC-LH-(αMeK)-L-Aib-DYPQTDVGAGS-Hyp-NH2;

[0248] U-ASHLSTA-Pen-LGRLSA-Pen-LH-(αMeK)-L-Aib-DYPQTDVGAGS-Hyp-NH2;

[0249] U-ASHLSTA-(αMeC)-LGRLSA-(αMeC)-LH-(αMeK)-L-Aib-DYPQTDVGAGS-Hyp-NH2;

[0250] U-ASHLSTACLGRLSACLH-(αMeK)-L-Aib-DYPQTDVGAGS-Hyp-NH2;

[0251] U-ASHLSTA-(N-Me-C)-LGRLSA-(N-Me-C)-LH-(αMeK)-L-Aib-DYPQTDVGAGS-Hyp-NH2;

[0252] U-ASHLSTAKLGRLSAKLH-(αMeK)-L-Aib-DYPQTDVGAGS-Hyp-NH2;

[0253] U-ASHLSTAVLGKLSAELHKL-Aib-DYPQTDVGAGS-Hyp-NH2;

[0254] U-ASHLSTA-Orn-LGRLSA-Orn-LH-(αMeK)-L-Aib-DYPQTDVGAGS-Hyp-NH2;

[0255] U-ASHLSTA-Dab-LGRLSA-Dab-LH-(αMeK)-L-Aib-DYPQTDVGAGS-Hyp-NH2;

[0256] Each U group is independently selected from H, acetyl, or benzoyl groups.

[0257] It should be noted that in the polypeptide structure shown in the above formula (I), the "-" between amino acids represents an amide bond; for example, "-" in "L-Aib-D" represents an amide bond.

[0258] According to an embodiment of the present invention, in the structure shown in formula (I), U is hydrogen.

[0259] According to an embodiment of the present invention, in the structure shown in formula (I), U is an acetyl group.

[0260] According to an embodiment of the present invention, in the structure shown in formula (I), U is benzoyl.

[0261] According to an embodiment of the present invention, the structure shown in formula (I) has at least one of the following table structures:

[0262]

[0263] It should be noted that in the tables of this article, when a polypeptide code corresponds to multiple polypeptides in Table A or Table B, it means that the polypeptide in the table has the same main peptide chain as multiple polypeptides in Table A or Table B, the only difference being that it does not have a modifying group side chain. For example, "1-21 (AMY-021, AMY-038)" means that polypeptide 1-21 in the above table has the same main peptide chain as AMY-021 and AMY-038 in Table A, but polypeptide 1-21 does not have a modifying group side chain; the same applies to "1-15 (AMY-015, AMY-016, AMY-017, AMY-018)".

[0264] According to embodiments of the present invention, the polypeptide or its derivatives or pharmaceutically acceptable salts further include modifying groups.

[0265] According to an embodiment of the present invention, the modifying group is linked to two amino acids Z1 or two amino acids Z2 in the polypeptide or its derivative represented by formula (I).

[0266] According to an embodiment of the present invention, the modifying group is connected to the -SH of the Z1 side chain of the amino acid via a sulfur-carbon bond.

[0267] According to an embodiment of the present invention, the modifying group is connected to the -NH2 of the Z2 side chain of the amino acid via an amide bond.

[0268] In an optional embodiment of the present invention, the modifying group is linked to the -SH of the C side chain of the amino acid via a sulfur-carbon bond.

[0269] In an optional embodiment of the present invention, the modifying group is connected to the ε-amino group of the K side chain of the amino acid via an amide bond.

[0270] According to an embodiment of the present invention, the modifying group has the structure shown in formula (IV):

[0271]

[0272] Where R1 is C, N, or -C 3～10 Heteroalkyl, -C 3～10 aryl or -C 3～10 heteroaryl;

[0273] R2 and R3 are each independently selected by one or more R... 1a Replacement -C 1～6 Alkylene-, selected by one or more R 1a Substituted -NH-C(O)-C 1～6 Alkylene - or optionally with one or more R 1a Replacement -C 1～6 Alkylene-NH-C(O)-C 1～6 alkylene-, wherein each R 1a Independently halogen, -OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl, -C 1～6 Halogenated alkyl or -C 1～6 Alkoxy;

[0274] R4 is empty, or can be arbitrarily assigned one or more R4 values. 2a Replacement -C 1～6 Alkylene-NH-C(O)-C 1～6 alkylene-, wherein each R 2a Independently halogen, -OH, -SH, -NH2, -NO2, -CN, phenyl, -C1～6 Alkyl, -C 1～6 Halogenated alkyl or -C 1～6 Alkoxy;

[0275] R5 represents H and -C. 1～6 alkoxy or -C 1～6 Alkyl, wherein the -C 1～6 Alkyl and -C 1～6 Each alkoxy group may be independently substituted by one or more halogens, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, or -CN;

[0276] R6 is arbitrarily assigned to one or more Rs 3a Replacement -C 1～6 Alkylene-, or optionally with one or more R 3a Replacement -(C 1～3 alkylene-O) m1 -C 1～6 alkylene-, wherein each R 3a Independently halogen, -OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl, -C 1～6 Halogenated alkyl or -C 1～6 Alkoxy;

[0277] R7 is arbitrarily assigned to one or more Rs 4a Replacement -C 1～6 alkylene-, wherein each R 4a Independently halogen, -OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl, -C 1～6 Halogenated alkyl or -C 1～6 Alkoxy;

[0278] R8 is arbitrarily assigned to one or more R... 5a Replacement -C 10～20 Alkyl, optionally with one or more R 5a Replacement -C 10～20 Alkylene-R9, optionally with one or more R 5a Replacement -C 5～10 Alkylene-OC 3～10 Aneryl-R9, or optionally one or more R 5a Replacement -C 5～10 Alkylene-OC 3～10 Hybrid aryl-R9, wherein each R 5a Independently halogen, -OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl, -C 1～6Halogenated alkyl or -C 1～6 Alkoxy;

[0279] R9 represents -COOH and -C. 3～7 heteroaryl, -S(O)2OH, or -PO(OH)2;

[0280] m1 is any integer from 1 to 6;

[0281] n1 is any integer from 0 to 6;

[0282] n2 is any integer from 1 to 10.

[0283] It should be noted that in this invention, the "-" in the modified group represents the chemical bond connecting chemical groups, such as the covalent bond between atoms (or atoms in a group) and atoms (or atoms in a group).

[0284] According to an embodiment of the present invention, R1 is C, N, or -C. 5～7 Heteroalkyl, -C 5～7 aryl or -C 5～7 Hybrid aryl.

[0285] According to embodiments of the present invention, R1 is N, phenylene, or pyridylene.

[0286] According to an embodiment of the present invention, R1 is

[0287] According to an embodiment of the present invention, R2 and R3 are each independently -C 1～6 alkylene - or -C 1～6 Alkylene-NH-C(O)-C 1～6 Alkylene-.

[0288] According to an embodiment of the present invention, R2 and R3 are each independently -C 1～3 alkylene - or -C 1～3 Alkylene-NH-C(O)-C 1～3 Alkylene-.

[0289] According to an embodiment of the present invention, each n1 is independently 0, 1, 2, 3, 4 or 5.

[0290] According to an embodiment of the present invention, n1 is 0.

[0291] According to an embodiment of the present invention, R4 is empty or -C 1～6 Alkylene-NH-C(O)-C 1～6 Alkylene-.

[0292] According to an embodiment of the present invention, R4 is empty or -C 1～3 Alkylene-NH-C(O)-C1～3 Alkylene-.

[0293] According to an embodiment of the present invention, R5 is H or -C. 1～3 alkoxy or -C 1～3 alkyl.

[0294] According to an embodiment of the present invention, R6 is -C 1～6 Alkylene-, -(C 1～3 alkylene-O) m1 -C 1～6 Alkylene-.

[0295] According to an embodiment of the present invention, R6 is -C 1～3 Alkylene-, -(C 1～3 alkylene-O) m1 -C 1～3 Alkylene-.

[0296] According to an embodiment of the present invention, m1 is 2, 3, 4 or 5.

[0297] According to an embodiment of the present invention, R7 is -C 1～6 Alkylene-.

[0298] According to an embodiment of the present invention, R7 is -C 2～4 Alkylene-.

[0299] According to an embodiment of the present invention, R8 is -C 10～20 Alkyl, -C 10～20 Alkylene-R9, -C 5～10 Alkylene-OC 3～10 aryl-R9 or -C 5～10 Alkylene-OC 3～10 Hybrid aryl-R9.

[0300] According to an embodiment of the present invention, R8 is -C 10～18 Alkyl, -C 14～18 Alkylene -R9, or -C 7～9 Alkylene-OC 5～7 aryl-COOH.

[0301] According to an embodiment of the present invention, R9 is -COOH or -C 5～6 heteroaryl, -S(O)2OH, or -PO(OH)2.

[0302] According to embodiments of the present invention, R9 is -COOH, -S(O)2OH, -PO(OH)2, or...

[0303] According to an embodiment of the present invention, n2 is 1, 2 or 3.

[0304] According to an embodiment of the present invention, n2 is 3, 4, 5 or 6.

[0305] According to an embodiment of the present invention, the modifying group represented by formula (IV) has the structure represented by formula (IVa):

[0306]

[0307] Among them, each R 10 and R 11 Each independently is -C 0～3 alkylene-;

[0308] R 12 -C 1～6 alkylene-;

[0309] R8 is -C 10～21 Alkyl, or -C 10～21 Alkylene-COOH, preferably -C 14～19 Alkyl or -C 10～18 alkylene-COOH;

[0310] q1 is 1, 2, 3, or 4;

[0311] q2 can be 1, 2, or 3.

[0312] According to an embodiment of the present invention, the modifying group represented by formula (IVa) has one of the following structures:

[0313]

[0314]

[0315]

[0316]

[0317] According to an embodiment of the present invention, the modifying group represented by formula (IV) has the structure represented by formula (IVb):

[0318]

[0319] Among them, each R 10 'and R 11 'Each is independent of -C 0～3 alkylene-;

[0320] R 12 'for-C 1～6 alkylene-;

[0321] R 13 -C 10～20Alkylene, preferably -C 14～18 alkylene-;

[0322] q1' is 1, 2, 3, or 4;

[0323] q2' can be 1, 2 or 3.

[0324] According to an embodiment of the present invention, the modifying group represented by formula (IVb) has one of the following structures:

[0325]

[0326] According to an embodiment of the present invention, the modifying group represented by formula (IV) has the structure represented by formula (IVc):

[0327]

[0328] Among them, each R 14 Each independently is -C 1～3 Alkylene- or -NH-C(O)-C 1～3 alkylene-;

[0329] R 15 -C 0～3 Alkylene- or -C(O)-NH-C 1～3 alkylene-;

[0330] R 12 "for -C 1～6 alkylene-;

[0331] R 13 'for-C 10～20 Alkylene, preferably -C 14～18 alkylene-;

[0332] "q1" can be 1, 2, 3, or 4;

[0333] "q2" can be 1, 2, or 3;

[0334] Y1 is either C or N.

[0335] According to an embodiment of the present invention, the modifying group represented by formula (IVc) has one of the following structures:

[0336]

[0337]

[0338] According to an embodiment of the present invention, the polypeptide or its derivative represented by formula (I) contains two of the amino acids Z1, and the modifying group has the structure represented by formula (IV).

[0339] According to an embodiment of the present invention, the modifying group has the structure shown in formula (V):

[0340]

[0341] Among them, R 20 For C, N, -C 3～10 aryl or -C 3～10 heteroaryl;

[0342] R 21 and R 22 Each is independently empty, and can be arbitrarily controlled by one or more R. 1b Replacement -C 1～6 Alkylene-, or optionally with one or more R 1b Replacement -C 1～6 alkylene oxide-, wherein each R 1b Independently halogen, -OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl, -C 1～6 Halogenated alkyl or -C 1～6 Alkoxy;

[0343] R 23 Empty, optional, can be one or more R 2b Replacement -C 1～6 Alkylene-, optionally with one or more R 2b Substituted -C(O)-C 1～6 Alkylene-, optionally with one or more R 2b Substituted -NH-C(O)-C 1～6 Alkylene-, or optionally with one or more R 2b Replacement -C 1～6 Alkylene-C(O)-NH-C 1～6 alkylene-, wherein each R 2b Independently halogen, -OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl, -C 1～6 Halogenated alkyl or -C 1～6 Alkoxy;

[0344] R 24 For optional use by one or more R 3b Replacement -C 1～6 Alkylene-, or optionally with one or more R 3b Replacement -(C 1～3 alkylene-O) m2 -C 1～6 alkylene-, wherein each R 3bIndependently halogen, -OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl, -C 1～6 Halogenated alkyl or -C 1～6 Alkoxy;

[0345] R 25 H, -C 1～6 alkoxy or -C 1～6 Alkyl, wherein the -C 1～6 Alkyl and -C 1～6 Each alkoxy group may be independently substituted by one or more halogens, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, or -CN;

[0346] R 26 For optional use by one or more R 4b Replacement -C 1～6 alkylene-, wherein each R 4b Independently halogen, -OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl, -C 1～6 Halogenated alkyl or -C 1～6 Alkoxy;

[0347] R 27 For optional use by one or more R 5b Replacement -C 10～20 Alkyl, optionally with one or more R 5b Replacement -C 10～20 Alkylene-R 28 Optional by one or more R 5b Replacement -C 5～10 Alkylene-OC 3～10 Aspartic-R 28 Or, arbitrarily selected by one or more R 5b Replacement -C 5～10 Alkylene-OC 3～10 Hybrid aryl-R 28 , among which, each R 5b Independently halogen, -OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl, -C 1～6 Halogenated alkyl or -C 1～6 Alkoxy;

[0348] R 28 -COOH, -C 3～7 heteroaryl, -S(O)2OH, or -PO(OH)2;

[0349] m2 is any integer from 1 to 6;

[0350] n3 is any integer from 1 to 10;

[0351] n3 is 0, 1, or 2;

[0352] Y2 is empty or NH.

[0353] According to an embodiment of the present invention, R 20 For C, N, -C 5～7 aryl or -C 5～7 Hybrid aryl.

[0354] According to an embodiment of the present invention, R 20 It is C, N, phenylene, or pyridylene.

[0355] According to an embodiment of the present invention, R 20 for Preferred

[0356] According to an embodiment of the present invention, R 21 and R 22 Each is independently empty, -C 1～6 alkylene - or -C 1～6 alkylene oxide-.

[0357] According to an embodiment of the present invention, R 21 and R 22 Each can be empty or -C independently. 1～3 Alkylene-.

[0358] According to an embodiment of the present invention, R 23 Empty, -C 1～6 alkylene-, -C 1～6 Alkylene-C(O)-NH-C 1～6 Alkylene- or -C(O)-C 1～6 Alkylene-.

[0359] According to an embodiment of the present invention, R 23 Empty, -C 1～3 Alkylene- or -C(O)-C 2～4 Alkylene-.

[0360] According to an embodiment of the present invention, R 24 -C 1～6 Alkylene - or -(C 1～3 alkylene-O) m2 -C 1～6 Alkylene-.

[0361] According to an embodiment of the present invention, R 24 -C 1～3Alkylene - or -(C 1～3 alkylene-O) m2 -C 1～3 Alkylene-.

[0362] According to an embodiment of the present invention, m2 is 2, 3, 4 or 5.

[0363] According to an embodiment of the present invention, R 25 H, -C 1～6 alkoxy or -C 1～6 alkyl.

[0364] According to an embodiment of the present invention, R 25 H or -C 1～3 alkyl.

[0365] According to an embodiment of the present invention, R 26 -C 1～6 Alkylene-.

[0366] According to an embodiment of the present invention, R 26 -C 1～3 Alkylene-.

[0367] According to an embodiment of the present invention, R 27 -C 10～20 Alkyl, -C 10～20 Alkylene-R 28 -C 5～10 Alkylene-OC 3～10 Aspartic-R 28 、or -C 5～10 Alkylene-OC 3～10 Hybrid aryl-R 28 .

[0368] According to an embodiment of the present invention, R 27 -C 10～18 Alkyl, -C 14～18 Alkylene-R 28 、or -C 7～9 Alkylene-OC 5～7 aryl-COOH.

[0369] According to an embodiment of the present invention, R 28 -COOH, -C 5～6 heteroaryl, -S(O)2OH, or -PO(OH)2.

[0370] According to an embodiment of the present invention, R 28 -COOH, -S(O)2OH, -PO(OH)2, or

[0371] According to an embodiment of the present invention, p is 1, 2 or 3.

[0372] According to an embodiment of the present invention, p is 3, 4, 5 or 6.

[0373] According to an embodiment of the present invention, the modifying group represented by formula (V) has the structure represented by formula (Va):

[0374]

[0375] Among them, each R 29 Each independently is -C 1～3 alkylene-;

[0376] R 30 -C(O)-C 1～6 alkylene - or -C 1～3 Alkylene-C(O)-NH-C 1～3 alkylene-;

[0377] R 31 -C 1～6 alkylene-;

[0378] q3 can be 1, 2, 3, or 4;

[0379] q4 can be 1, 2, or 3.

[0380] According to an embodiment of the present invention, in formula (Va), R 27 -C 10～20 alkylene -COOH or -C 10～20 Alkyl group, preferably -C 14～18 alkylene -COOH or -C 10～19 alkyl.

[0381] According to an embodiment of the present invention, the modifying group represented by formula (Va) has the following structure:

[0382]

[0383]

[0384] According to an embodiment of the present invention, the modifying group represented by formula (V) has the structure represented by formula (Vb):

[0385]

[0386] Among them, each R 29 'Each is independent of -C 1～3 alkylene-;

[0387] R 30 'for-C 1～6 alkylene-;

[0388] R 31 'for-C 1～6 alkylene-;

[0389] q3' can be 1, 2, 3 or 4, preferably 2;

[0390] q4' can be 1, 2 or 3, preferably 2.

[0391] According to an embodiment of the present invention, in formula (Vb), R 27 -C 10～20 alkylene -COOH or -C 10～20 Alkyl group, preferably -C 14～18 Alkylene-COOH. According to embodiments of the present invention, the modifying group represented by formula (Vb) has the following structure:

[0392]

[0393] According to an embodiment of the present invention, the modifying group represented by formula (V) has the structure represented by formula (Vc):

[0394]

[0395] Among them, R 30 "for -C 1～6 alkylene-;

[0396] R 31 "for -C 1～6 alkylene-;

[0397] q3 can be 1, 2, 3, or 4;

[0398] q4 can be 1, 2, or 3.

[0399] According to an embodiment of the present invention, in formula (Vc), R 27 -C 10～20 alkylene -COOH or -C 10～20 Alkyl group, preferably -C 14～18 alkylene -COOH or -C 10～19 alkyl.

[0400] According to an embodiment of the present invention, the modifying group represented by formula (Vc) has the following structure:

[0401]

[0402] According to an embodiment of the present invention, the polypeptide or its derivative represented by formula (I) contains two of the amino acids Z2, and the modifying group has the structure represented by formula (V).

[0403] According to embodiments of the present invention, the polypeptide derivative has any of the structures shown in the following table:

[0404]

[0405]

[0406] According to embodiments of the present invention, the polypeptide derivative has the structure shown in either Table A or Table B:

[0407] Table A

[0408]

[0409] It should be noted that, based on Table A, replacing the two amino acids linked to the modifying groups in peptides AMY-001~AMY-020 and AMY-023~AMY-036 with K, and replacing the modifying groups with the modifying groups shown in formula (V) (e.g., B1), yields peptide sequences with performance similar to peptides AMY-001~AMY-020 and AMY-023~AMY-036 in Table A. Therefore, this invention does not show all peptides; all are within the scope of protection of this application. This invention only exemplarily shows some peptides; see Table B for details. Specific activity data can be found in the Examples and Test Examples section of this invention.

[0410] Table B

[0411]

[0412] It should be noted that the "linking sites of the modifying groups" in Tables A and B specifically refer to X in the structure shown in formula (I) of this invention. i The position of i in the equation.

[0413] Pharmaceutical compositions, combination drugs or kits

[0414] In a second aspect, the present invention provides a pharmaceutical composition. According to embodiments of the invention, the pharmaceutical composition comprises the polypeptide or its derivatives or pharmaceutically acceptable salts described in the first aspect. As is previously known, polypeptides or their derivatives or pharmaceutically acceptable salts (hereinafter referred to as polypeptides or analogs) are dual-agonist polypeptide analogs, which can simultaneously possess a certain degree of balanced or unbalanced activation activity against both the amylin receptor (AMYR) and the calcitonin receptor (CTR). Therefore, the use of a drug containing polypeptides or their derivatives or pharmaceutically acceptable salts can effectively prevent or treat diseases related to the amylin receptor and / or calcitonin receptor.

[0415] According to embodiments of the present invention, the pharmaceutical composition further comprises pharmaceutically acceptable excipients or carriers.

[0416] In a third aspect, the present invention provides a combination drug or kit. According to embodiments of the invention, the combination drug or kit comprises: a polypeptide or derivative thereof described in the first aspect, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described in the second aspect as a first active ingredient; and a second active ingredient; wherein the second active ingredient comprises a drug for the prevention and / or treatment of a disease. The combination drug or kit according to embodiments of the invention can further improve the therapeutic effect on diseases related to amylin receptors and / or calcitonin receptors.

[0417] According to embodiments of the present invention, the disease includes prevention and / or treatment of diseases related to amylin receptor and / or calcitonin receptor.

[0418] According to embodiments of the present invention, the amylin receptor and / or calcitonin receptor-related diseases include at least one of lipid metabolism disorders, glucose metabolism disorders, cardiovascular diseases, brain diseases, mental diseases, or nervous system diseases.

[0419] According to embodiments of the present invention, the amylin receptor and / or calcitonin receptor-related diseases include at least one of metabolic disorder-related diseases, bone-related diseases, cardiovascular diseases unrelated to metabolic diseases, symptoms or diseases related to alcohol or drug addiction, and neurodegenerative diseases.

[0420] According to embodiments of the present invention, the amylin receptor and / or calcitonin receptor-related diseases include at least one of diabetes, hypertension, arteriosclerosis, cirrhosis, coronary heart disease, angina pectoris, myocardial infarction, inflammatory bowel disease, dyspepsia, gastrointestinal ulcers, hyperglycemia, impaired glucose tolerance, syndrome X, cognitive impairment, stroke, dyslipidemia-related diseases (hyperlipidemia, dyslipidemia), metabolic syndrome, non-alcoholic steatohepatitis, non-alcoholic fatty liver disease, abnormal weight, obesity, fatty liver disease, diabetic nephropathy syndrome, diabetes-associated renal fibrosis, liver fibrosis, Alzheimer's disease, and Parkinson's disease.

[0421] According to embodiments of the present invention, the amylin receptor and / or calcitonin receptor-related diseases include at least one of cardiovascular disease, diabetes, and / or obesity.

[0422] According to embodiments of the present invention, the drug is selected from at least one of cardiovascular, diabetes and / or obesity drugs.

[0423] According to embodiments of the present invention, the drug is selected from at least one of the following: adrenergic receptor blockers, HMG-CoA reductase inhibitors, angiotensin receptor antagonists, angiotensin-converting enzyme inhibitors, calcium channel blockers, endothelin antagonists, renin inhibitors, diuretics, aldosterone receptor blockers, endothelin receptor blockers, aldosterone synthase inhibitors, CETP inhibitors, relaxin, PCSK9 inhibitors, BNP and NEP inhibitors, GLP-1 analogs, insulin, sulfonylureas, biguanides, meglitazones, glucosidase inhibitors, DPP IV inhibitors, and SGLT2 inhibitors.

[0424] According to embodiments of the present invention, the GLP-1 analog drug includes at least one of Semaglutide, Exenatide, Liraglutide, Dulaglutide, Tirzepatide, and Retatrutdie.

[0425] According to an embodiment of the present invention, the glibenclamide-type hypoglycemic drug includes at least one of dapagliflozin and empagliflozin.

[0426] According to an embodiment of the present invention, the biguanide class includes metformin.

[0427] Uses and methods

[0428] In a fourth aspect of the invention, the invention provides for the use of the polypeptides or derivatives thereof or pharmaceutically acceptable salts thereof described in the first aspect, the pharmaceutical compositions described in the second aspect, or the combination pharmaceuticals or cassettes described in the second aspect in the preparation of a medicament for the treatment or prevention of diseases related to amylin receptors and / or calcitonin receptors.

[0429] According to embodiments of the present invention, the diseases related to the amylin receptor and / or calcitonin receptor include lipid metabolism disorders, glucose metabolism disorders, cardiovascular diseases, brain diseases, mental diseases, or nervous system diseases.

[0430] According to embodiments of the present invention, the amylin receptor and / or calcitonin receptor-related diseases include at least one of metabolic disorder-related diseases, bone-related diseases, cardiovascular diseases unrelated to metabolic diseases, symptoms or diseases related to alcohol or drug addiction, and neurodegenerative diseases.

[0431] According to embodiments of the present invention, the amylin receptor and / or calcitonin receptor-related diseases include at least one of diabetes, hypertension, arteriosclerosis, cirrhosis, coronary heart disease, angina pectoris, myocardial infarction, inflammatory bowel disease, dyspepsia, gastrointestinal ulcers, hyperglycemia, impaired glucose tolerance, syndrome X, cognitive impairment, stroke, dyslipidemia-related diseases (hyperlipidemia, dyslipidemia), metabolic syndrome, non-alcoholic steatohepatitis, non-alcoholic fatty liver disease, abnormal weight, obesity, fatty liver disease, diabetic nephropathy syndrome, diabetes-associated renal fibrosis, liver fibrosis, Alzheimer's disease, and Parkinson's disease.

[0432] In a fifth aspect, the present invention provides a method for preventing and / or treating diseases related to the amylin receptor and / or calcitonin receptor. According to embodiments of the invention, the method comprises administering to a subject a pharmaceutically acceptable amount of the polypeptide or its derivatives or pharmaceutically acceptable salts thereof described in the first aspect, the pharmaceutical composition described in the second aspect, or the combination drug or kit described in the second aspect. As is known prior art, the polypeptide or its derivatives or pharmaceutically acceptable salts thereof (hereinafter referred to as polypeptides or analogues) are dual-agonist polypeptide analogues, which may simultaneously possess a certain balanced or unbalanced activation activity against both the amylin receptor (AMYR) and the calcitonin receptor (CTR). Therefore, the method of the present invention can effectively prevent and / or treat diseases related to the amylin receptor and / or calcitonin receptor.

[0433] According to embodiments of the present invention, the amylin receptor and / or calcitonin receptor-related diseases include at least one of metabolic disorder-related diseases, bone-related diseases, cardiovascular diseases unrelated to metabolic diseases, symptoms or diseases related to alcohol or drug addiction, and neurodegenerative diseases.

[0434] According to embodiments of the present invention, the metabolic disorder-related diseases include at least one of type 2 diabetes, dyslipidemia-related diseases, metabolic syndrome, non-alcoholic steatohepatitis, non-alcoholic fatty liver disease, obesity, fatty liver disease, and so on.

[0435] According to embodiments of the present invention, the neurodegenerative disease includes at least one of Alzheimer's disease and Parkinson's disease.

[0436] The effective amount of the polypeptides or their derivatives, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions described in this invention may vary depending on the mode of administration and the severity of the disease to be treated. A preferred effective amount can be determined by those skilled in the art based on various factors (e.g., through clinical trials). These factors include, but are not limited to: pharmacokinetic parameters of the active ingredient, such as bioavailability, metabolism, and half-life; the severity of the disease to be treated, the patient's weight, the patient's immune status, and the route of administration. For example, due to the urgency of the treatment condition, several separate doses may be administered daily, or the dose may be reduced proportionally.

[0437] The polypeptides or derivatives thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions of the present invention, may be incorporated into medicaments suitable for parenteral administration (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). These medicaments may be prepared in various forms, such as liquids, semi-solids, and solid dosage forms, including but not limited to liquid solutions (e.g., injection solutions and infusion solutions) or lyophilized powders. Typical medicaments are in the form of injection solutions or infusion solutions. The aforementioned polypeptides or derivatives thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions may be administered by intravenous infusion or injection, or by intramuscular or subcutaneous injection.

[0438] The present invention will be explained below with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained commercially.

[0439] Example 1: Preparation method of polypeptide or its derivative

[0440] 1. The linear peptides in peptides 1 to 20 (abbreviated as AMY-001 to AMY-020) and peptides 23 to 36 (abbreviated as AMY-023 to AMY-031) in Table A were synthesized using the classic Fmoc-tBu solid-phase synthesis method. Prepared using an X-peptide synthesizer, under the following reaction conditions:

[0441] (1) Resin swelling: Add Rink Amide MBHA resin to DCM (dichloromethane), react with N2 gas at room temperature for 1 hour, filter, and wash the resin with DMF 2-3 times.

[0442] (2) Removal of Fmoc protecting group: Add a DMF (N,N-dimethylformamide) solution containing 20% ​​(v / v) piperidine to the above resin, react with N2 gas at room temperature for 10 minutes, filter, repeat the above operation until the protection is completely removed, and wash the resin with DMF 2-3 times.

[0443] (3) Amino acid coupling: The reactants were fed according to a reaction ratio of resin: amino acid: DIC: Oxyma Pure = 1:5:5:5 (equivalent ratio). The reactants, DIC (N,N'-diisopropylcarbodiimide), and Oxyma Pure (condensation reagent) were pre-dissolved in DMF. The reaction was carried out under N2 atmosphere at room temperature for 10 minutes, and then transferred to a resin reaction vessel. The reaction was carried out under N2 atmosphere at room temperature for 1-3 hours. After the reaction was completed, the mixture was filtered, and the resin was washed 2-3 times with DMF. After the synthesis was completed, the resin was washed 2-3 times with DCM and dried under vacuum to obtain the peptide resin.

[0444] The above method is applicable to, but is not limited to, the following amino acids (D or L type) or synthetic reagents: Fmoc-AEEA-OH, Fmoc-Aib-OH, Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Asp(tBu)-OH, Fmoc-Cys(Trt)-OH, Fmoc-Gly-OH, Fmoc-Glu(tBu)-OH, Fmoc-Gln(Trt)-OH, Fmoc-His(Trt)-OH, Fmoc-Ile-OH, Fmoc-Leu-OH, Fmoc-Lys(Boc)-OH, Fmoc-Lys(Mtt)-OH, Fmoc-Lys(Alloc)-OH, Fmoc-Orn(Boc)-OH, Fmoc-Pro-OH, Fmoc-P he-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Val-OH, Fmoc-α-methyl-Leu-OH, Fmoc-α-methyl-Lys(Boc)-OH, Fmoc-α-methyl -Phe-OH, Fmoc-N-Me-Arg(Pbf)-OH, Fmoc-homoArg(Boc)-OH, Fmoc-cys(Trt)-OH, Fmoc-arg(Pbf)-OH, Fmoc-Pro(4-S-NH-Boc)-OH, Fmoc-Pro(4-R-NH-Boc)-OH, Fmoc-pro-OH, etc.

[0445] 2. Linear peptide cleavage and drying: Prepare the peptide cleavage buffer according to the volume ratio of TFA (trifluoroacetic acid): TIPS (triisopropylsilylethynyl): H2O: EDT (1,2-ethylenedithiol) = 95:2:2:1. Add the cleavage buffer (10 mL / g resin) to the dried peptide resin, shake thoroughly for 3 hours, filter the resin residue, add 10 times the volume of cold MTBE (methyl tert-butyl ether) to the filtrate, cool the resulting suspension at -20℃ for 1 hour, then centrifuge at 3500 rpm, wash the precipitate 3-5 times with cold MTBE, and vacuum dry to obtain the crude peptide.

[0446] 3. Purification of linear peptides: A solution was prepared according to the volume ratio of mobile phase A (0.1% (v / v) TFA (trifluoroacetic acid) - water) : mobile phase B (0.1% (v / v) TFA - acetonitrile) = 1.5 : 1. The crude peptide was dissolved in the solution to prepare a stock solution. The solution was filtered through a 0.45 μm filter membrane. A C18 reversed-phase preparative column (20*250 mm, 5 μm particle size) was used for gradient elution with mobile phases A and B at a flow rate of 10 mL / min. The target peak was collected and lyophilized to obtain the target peptide compound (i.e., the linear peptide).

[0447] 4. The Staple linker (or modifying group A) was prepared using the classic Fmoc-tBu solid-phase synthesis method. This step takes Staple-A1 (or modifying group A1) as an example, and the reaction conditions are shown below (see details of the synthesis process). Figure 1 ):

[0448] (1) Resin swelling: Add Fmoc-L-Lys(ivDde)-2CTC resin to DCM, react with N2 gas at room temperature for 1 hour, filter, and wash the resin with DMF 2-3 times.

[0449] (2) Removal of Fmoc protecting group: Add DMF solution containing 20% ​​piperidine to the above resin, react with N2 gas at room temperature for 10 minutes, filter, repeat the above operation until complete deprotection, and wash the resin with DMF 2-3 times.

[0450] (3) Fatty acid chain coupling: The resin, fatty acid, HATU (2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate), and DIPEA (N,N-diisopropylethylamine) were fed in a reaction ratio of 1:5:5:10 (volume ratio). The fatty acid monotert-butyl ester, HATU and DIPEA were pre-dissolved in DMF and reacted with N2 at room temperature for 10 minutes. Then the mixture was transferred to a resin reaction vessel and reacted with N2 at room temperature for 1-3 hours. After the reaction was completed, the mixture was filtered and the resin was washed with DMF 2-3 times.

[0451] (4) Removal of ivDde protecting group: Add DMF solution containing 5% hydrazine to the resin, react with N2 gas at room temperature for 10 minutes, repeat this operation 2-3 times until completely removed, and wash the resin with DMF 2-3 times.

[0452] (5) Coupling of amino acids, AEEA, and bromoacetic acid: Reactants were added according to a reaction equivalence ratio of resin:Fmoc-AA-OH:DIC:Oxyma Pure = 1:5:5:5. The reactants, DIC, and Oxyma Pure were pre-dissolved in DMF and reacted under N2 atmosphere at room temperature for 10 minutes. The mixture was then transferred to a resin reaction vessel and reacted under N2 atmosphere at room temperature for 1-3 hours. After the reaction, the mixture was filtered, and the resin was washed 2-3 times with DMF. After synthesis, the resin was washed 2-3 times with DCM and dried under vacuum to obtain the linker resin.

[0453] The structure of the Staple linker can be the above-mentioned modifying groups A1 to A22, and can be selected from the following structures for example:

[0454]

[0455]

[0456] 5. Coupling of linear peptides to Staple linkers:

[0457] Dissolve 1.0 equivalent of the linear peptide (i.e., the target peptide compound obtained in step 3, taking peptide DAC09 (abbreviated as AMY-008) in Table A as an example) and 1.2 equivalent of the Staple linker peptide (i.e., the Staple linker obtained in step 4) in a reaction solution of PBS (phosphate buffered saline):acetonitrile = 1:1.5 (final concentration 1.2 mM). Adjust the pH of the reaction solution to 8.0 with 10% (v / v) NaOH aqueous solution. Shake the solution thoroughly at room temperature for 3-8 hours until the linear peptide is completely consumed as detected by LC-MS. Neutralize the reaction solution to pH 8.0 with 5% (v / v) TFA aqueous solution. 6.5. Filter using a 0.45 μm filter membrane, and perform gradient elution using a C18 reverse-phase column (20*250 mm, 5 μm particle size) at a flow rate of 10 mL / min with mobile phases A and B. Collect the target peak, freeze-dry, and obtain the target conjugated compound (i.e., peptide derivatives, AMY-001~AMY-020, AMY-023~AMY-031). See the following formula for the specific synthesis process:

[0458]

[0459] The purity of each polypeptide derivative was determined by mass spectrometry. HPLC analysis showed that the purity of all derivatives was greater than 90%. Mass spectrometry analysis confirmed that the molecular weights of the polypeptides were basically consistent with the theoretical molecular weights (within the allowable error range). This embodiment exemplarily shows the molecular weights of some polypeptide molecules, as detailed in Table 1.

[0460] Table 1

[0461]

[0462] Example 2: Preparation method of polypeptide or its derivative

[0463] 1. The linear peptides in peptides 21-22 (abbreviated as AMY-021-AMY-022) and 37-40 (abbreviated as AMY-037-AMY-041) in Table A were synthesized using the classic Fmoc-tBu solid-phase synthesis method. Prepared using an X-peptide synthesizer, under the following reaction conditions:

[0464] (1) Resin swelling: Add Rink Amide MBHA resin to DCM, react with N2 gas at room temperature for 1 hour, filter, and wash the resin with DMF 2-3 times.

[0465] (2) Removal of Fmoc protecting group: Add a DMF solution containing 20% ​​(volume percentage, v / v) piperidine to the above resin, react with N2 gas at room temperature for 10 minutes, filter, repeat the above operation until complete deprotection, and wash the resin with DMF 2-3 times.

[0466] (3) Amino acid coupling: The resin, Fmoc-AA-OH, DIC, and Oxyma Pure were fed in a reaction ratio of 1:5:5:5. Fmoc-AA-OH, DIC, and Oxyma Pure were pre-dissolved in DMF and reacted under N2 atmosphere at room temperature for 10 minutes. The mixture was then transferred to a resin reaction vessel and reacted under N2 atmosphere at room temperature for 1-3 hours. After the reaction, the mixture was filtered, and the resin was washed 2-3 times with DMF. Amino acids with N-terminal sequences such as Boc-L-His(Trt)-OH and Boc-L-Tyr(tBu)-OH were synthesized using the same method. After synthesis, the resin was washed 2-3 times with DCM and dried under vacuum to obtain a peptide resin containing linear peptides.

[0467] The above method is applicable to, but is not limited to, the following amino acids (D or L type) or synthetic reagents: Fmoc-AEEA-OH, Fmoc-Aib-OH, Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Asp(tBu)-OH, Fmoc-Cys(Trt)-OH, Fmoc-Gly-OH, Fmoc-Glu(tBu)-OH, Fmoc-Gln(Trt)-OH, Fmoc-His(Trt)-OH, Fmoc-Ile-OH, Fmoc-Leu-OH, Fmoc-Lys(Boc)-OH, Fmoc-Lys(Mtt)-OH, Fmoc-Lys(Alloc)-OH, Fmoc-Orn(Boc)-OH, Fmoc-Pro-OH, Fmoc- Phe-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Val-OH, Fmoc-α-methyl-Leu-OH, Fmoc-α-methyl-Lys(Boc)-OH, Fmoc-α-methy l-Phe-OH, Fmoc-N-Me-Arg(Pbf)-OH, Fmoc-homoArg(Boc)-OH, Fmoc-cys(Trt)-OH, Fmoc-arg(Pbf)-OH, Fmoc-Pro(4-S-NH-Boc)-OH, Fmoc-Pro(4-R-NH-Boc)-OH, Fmoc-pro-OH, etc.

[0468] 2. The connection between the Staple linker (or modifying group B) and the peptide resin containing the linear peptide (AMY-022) obtained in step 1 is as follows:

[0469]

[0470] 2.1 Removal of Mtt protecting group from peptide resin: Prepare a deprotection solution according to the volume ratio of TFA:TIPS:DCM = 1:2:97. Add Mtt removal solution (10 mL / g resin) to the resin, place it on a shaker and shake it thoroughly for 15 minutes. Filter the resin and repeat the process 3-5 times until the addition of the removal solution no longer shows a bright yellow color indicating complete removal. Wash the resin with DMF 2-3 times.

[0471] 2.2 Removal of Alloc and Allyl protecting groups from peptide resin: Add DCM (10 mL / g resin), PhSiH3 (10 equivalents), and Pd(PPh3)4 (0.1 equivalents) to the resin, react under N2 atmosphere at room temperature for 1 hour, filter the resin, repeat the process 1-2 times, and wash the resin 2-3 times with DMF.

[0472] 2.3 Cyclone closure reaction on peptide resin: DMF (10 mL / g resin), PyBOP (5 equivalents), HOBt (5 equivalents), and DIPEA (10 equivalents) were added to the resin after removing the Alloc and Allyl protecting groups. The reaction was carried out under N2 atmosphere at room temperature for 4 hours. The resin was then filtered. This process was repeated 1-2 times. The resin was washed 2-3 times with DMF.

[0473] 2.4 Introduction of solid-phase side chains: The process was carried out according to the standard solid-phase synthesis method, wherein the side chains have the following structure:

[0474]

[0475] Furthermore, the structure of the Staple linker can be the aforementioned modifying groups B1 to B23.

[0476] 3. Cutting and Drying: Prepare the peptide cutting solution according to the volume ratio of TFA:TIPS:H2O:EDT = 95:2:2:1. Add the cutting solution (10mL / g resin) to the dried resin, shake thoroughly on a shaker for 3 hours, filter the resin residue, add 10 times the volume of cold MTBE to the filtrate, cool the resulting suspension at -20℃ for 1 hour, then centrifuge at 3500rpm, wash the precipitate 3-5 times with cold MTBE, and vacuum dry to obtain the crude peptide.

[0477] 4. Peptide Purification: A solution was prepared using mobile phase A (0.1% TFA-water) : mobile phase B (0.1% TFA-acetonitrile) at a volume ratio of 1.5 : 1. The crude peptide was dissolved in the solution to prepare a stock solution. The solution was filtered through a 0.45 μm filter membrane. A C18 reversed-phase preparative column (20*250 mm, 5 μm particle size) was used for gradient elution with mobile phases A and B at a flow rate of 10 mL / min. The target peak was collected, and the target peptide derivatives, namely AMY-021 to AMY-022, were obtained by lyophilization. The purity of each peptide derivative was determined by HPLC and mass spectrometry. The purity of all peptides was greater than 90%. The molecular weights of the peptides determined by mass spectrometry were basically consistent with the theoretical molecular weights (within the allowable error range). The molecular weights of some peptide molecules are shown in Table 2 in this example.

[0478] Table 2: Molecular weight of compounds

[0479]

[0480] Test Example 1: In vitro activity assay

[0481] 1. The polypeptide derivatives prepared in Examples 1 and 2 of this invention can bind to the target AMYR and CTR receptors on the cell membrane, activating the cAMP response element (CRE) and initiating the expression of downstream luciferase. The expression level is positively correlated with the biological activity of the tested analog. After activation, luciferase substrate is added for chemiluminescence detection, and the luminescence intensity is measured to characterize the biological activity of the tested compound. Therefore, this invention detects the polypeptide derivatives prepared in Examples 1 and 2, and the specific steps are as follows:

[0482] A stable HEK293 / pGM-CREB-L-Luc / AMY3R pooled transfection cell line was constructed. The polypeptide derivatives (AMY-001 to AMY-041) prepared in Examples 1 and 2 were diluted 5-fold from a maximum of 1000 nM to prepare a dilution series containing 8-11 concentration gradients. Cagrilintide was selected as a positive control, and a dilution series was prepared for testing using the same method. The corresponding test cells were digested (1 min), centrifuged (1000 rpm, 5 min), the culture medium was discarded, and the cells were resuspended in freestyle medium, centrifuged, collected, counted, and diluted to a cell density of 5*102. 5 Add 40 μL / 20,000 cells / well to the test wells of a 384-well plate. Use Echo to add the corresponding analyte dilution series. Incubate the cell plate at 37°C and 5% CO2 for 6 hours, then add ONE-GLO (20 μL / well) for detection. Incubate in the dark for 3 minutes, then perform measurements using a chemiluminescent microplate reader. Read the plate within 30 minutes and record the results. Use GraphPad Prism software to plot activation curves and calculate the EC50 of the compounds. 50 Values. The results show that the polypeptide derivatives of the present invention have binding activity with AMYR and CTR.

[0483] 2. The polypeptide derivatives prepared in Examples 1 and 2 of this invention can bind to the target AMYR and CTR receptors on the cell membrane, activate the receptors, and release cAMP. Therefore, the activity of the analyte can be determined by using a cAMP detection kit and the HTRF (homogeneous time-resolved fluorescence) method. The specific steps are as follows:

[0484] Stable transfection cell lines were constructed using the HEK293 / pGM-CREB-L-Luc / AMY3R pool and the HEK293 / pGM-CREB-L-Luc / CTR pool. The polypeptide derivatives (AMY-001 to AMY-041) prepared in Examples 1 and 2 were diluted 5-fold starting from a maximum of 1000 nM to prepare a dilution series containing 8-11 concentration gradients. Cagrilintide and sCT were selected as positive controls, and dilution series were prepared for testing using the same method. The test cell lines were digested with cAMP-specific trypsin, resuspended in 3 mL of serum-free DMEM medium, and then counted. Add 5 μL IBMX (prepared with serum-free DMEM medium, 0.5 mM) and test cells (5 μL / 7500 cells / well) sequentially to 384-well plates. Incubate at 37°C and 5% CO2 for 30 minutes, then add cAMP-d2 (5 μL, 1x) and Anti-cAMP-Cryptate (5 μL, 1x). Incubate at room temperature for 1 hour. Measure the HTRF value using a chemiluminescent microplate reader and record the results. Plot activation curves using GraphPad Prism software and calculate the IC50 of the peptide derivative. 50 The results indicate that the polypeptide derivatives of the present invention exhibit binding activity with AMY3R and CTR. This embodiment exemplarily demonstrates the IC50 values ​​of some polypeptides. 50 Values, as detailed in Table 3. Figure 2 and Figure 3 As shown.

[0485] Table 3: IC50 of each target peptide derivative 50 value

[0486]

[0487] Test Example 2: Pharmacokinetic Evaluation

[0488] 1. The pharmacokinetic behavior of the polypeptide derivatives (i.e., AMY-001 to AMY-041) prepared in Examples 1 and 2 was tested in SD rats.

[0489] Male SD rats were administered each polypeptide derivative (AMY-001 to AMY-041) via single injection, either subcutaneously (SC, 3 mg / kg, PBS solvent, concentration 1.5 mg / mL, administration volume 2 mL) or intravenously (IV, 1 mg / kg, PBS solvent, concentration 0.5 mg / mL, administration volume 2 mL). Whole blood samples were collected from the jugular vein or other suitable veins at selected time points after administration. 0.2 mL of blood was placed in labeled EDTA-K2 anticoagulant tubes. After gently inverting the tubes to thoroughly mix the anticoagulant (EDTA-K2) with the blood, the tubes were immediately placed on wet ice and centrifuged as quickly as possible to separate the plasma. Centrifugation conditions were set at 4°C, 6800 g, and 6 minutes. Plasma samples were stored at a temperature not exceeding -20°C for analysis.

[0490] Under ice-water bath and yellow light conditions, (1) except for the blank sample, 300 μL of precipitant containing internal standard was added to a 96-well plate containing 20 μL of standard curve sample, quality control sample, or unknown sample; 300 μL of acetonitrile was added to the blank sample; (2) vortexed; (3) centrifuged; (4) 150 μL of supernatant was transferred to a new 96-well plate, and 150 μL of ultrapure water was added and mixed. (5) The above samples were quantitatively analyzed by LC-MS. Gradient elution was performed using XB-C18 (2.1*100mm, 3μm) chromatography. The mobile phase used was: mobile phase A = 0.1% (v / v) formic acid-water; mobile phase B = acetonitrile. Pharmacokinetic parameters were calculated using PhoenixWinNonlin software. The results showed that the polypeptide derivatives of the present invention were well absorbed in rats, with high blood drug concentrations and long half-lives. The pharmacokinetic parameters of some polypeptide derivatives are shown in Table 4 and Figure 4.

[0491] Table 4: Rat PK data for some polypeptide derivatives

[0492]

[0493] 2. The pharmacokinetic behavior of the polypeptide derivatives (i.e., AMY-001 to AMY-041) prepared in Examples 1 and 2 was tested in cynomolgus monkeys.

[0494] Adult male cynomolgus monkeys were administered each polypeptide derivative (AMY-001 to AMY-041) via single injection, either subcutaneously (SC, 0.2 mg / kg, PBS solvent, concentration 0.2 mg / mL, administration volume 1 mL) or intravenously (IV, 1 mg / kg, PBS solvent, concentration 0.2 mg / mL, administration volume 1 mL). Whole blood samples were collected from the jugular vein or other suitable veins at selected time points after administration. 0.2 mL of blood was placed in labeled EDTA-K2 anticoagulant tubes. After gently inverting the tubes to thoroughly mix the anticoagulant (EDTA-K2) with the blood, the tubes were immediately placed on wet ice and centrifuged as quickly as possible to separate the plasma. Centrifugation conditions were set at 4°C, 6800 g, and 6 minutes. Plasma samples were stored at a temperature not exceeding -20°C for analysis.

[0495] Under ice-water bath yellow light conditions, (1) except for the blank sample, add 300 μL of precipitant containing internal standard to a 96-well plate containing 20 μL of standard curve sample, quality control sample or unknown sample; add 300 μL of acetonitrile to the blank sample; (2) vortex mix; (3) centrifuge; (4) take 150 μL of supernatant to a new 96-well plate, and then add 150 μL of ultrapure water and mix. (5) Quantitatively analyze the above samples using LC-MS analysis, using XB-C18 (2.1*100mm, 3μm) chromatography with gradient elution. The mobile phase used is: mobile phase A = 0.1% formic acid-water; mobile phase B = acetonitrile. Pharmacokinetic parameters were calculated using Phoenix WinNonlin software. The results showed that the polypeptide derivatives of the present invention were well absorbed in rats, with high blood drug concentrations and long half-lives. The pharmacokinetic parameters of some polypeptide derivatives are shown in Table 5 and Figure 5 As shown.

[0496] Table 5: Comparison data of some polypeptide derivatives in cynomolgus monkeys

[0497]

[0498] Test Example 3: Changes in body weight and food intake in wild-type SD rats after a single drug administration

[0499] Adult male wild-type SD rats, weighing 200-250g, were used in the experiment. The SD rats were randomly divided into groups of four, with two rats per cage, and were acclimatized for one week, during which they were given a normal diet and free access to water. After the animal acclimatization, the basal blood glucose and body weight of each rat were measured before drug administration (Day 0). Subsequently, according to the experimental groups, the rats were administered the solvent control (Vehicle) and three doses of the polypeptide derivatives (i.e., AMY-001 to AMY-041) prepared in Examples 1 and 2 of this invention (3 nmol / kg, 10 nmol / kg, and 30 nmol / kg) via subcutaneous injection (sc, "sc" and "SC" are synonymous). After the drug administration, the rats were given a normal diet and water. On Days 1, 2, 3, 4, 5, 6, and 7 after drug administration, the body weight of each rat and the total daily food intake of each group were measured. The body weight (% of initial value) and the cumulative food intake change of each group were calculated, and curves were plotted. The results showed that the compound of this invention can reduce the body weight and food intake of rats. This test case demonstrates the results for AMY-015. (See [link to test case]) Figure 6 Test Example 4: Changes in body weight, blood glucose, and food intake in a DIO rat model after continuous drug administration.

[0500] Adult male Sprague Dawley rats, weighing 200-250g, were used in the experiment and fed a high-fat diet for 16 weeks, reaching a weight of approximately 600-700g. These rats were randomly divided into groups of five and housed individually for one week of acclimatization, during which they received a normal diet and free access to water. After the acclimatization period, the baseline weight of each rat was measured before administration (Day 0). Subsequently, the rats were administered subcutaneously (sc) the following drugs: a solvent control (Vehicle), Cagrilintide (1 nmol / kg, 3 nmol / kg, 10 nmol / kg, Q2D), and AMY-015 (1 nmol / kg, 3 nmol / kg, 10 nmol / kg, Q2D). After the administration of these drugs, the rats were given a normal high-fat diet and water. On days 1, 2, 3, 4, 5, 6, and 7 after drug administration, the body weight of each rat and the total daily food intake of each group were measured. The changes in body weight (% of initial value) and total daily food intake of each group were calculated, and curves were plotted. The results show that the compound of the present invention can reduce the body weight and food intake of rats. The results of AMY-015 are exemplarily shown in this test example; see [link to relevant documentation]. Figure 7 .

[0501] Test Example 5: Changes in body weight and adipose tissue in DIO rat models after continuous administration of the drug alone or in combination with semaglutide.

[0502] Adult male Sprague Dawley rats, weighing 200-250g, were used in the experiment and fed a high-fat diet for 20 weeks until they reached a weight of approximately 600-700g. These rats were then randomly divided into groups of seven, and kept in solitary cages for one week for acclimatization, during which time they received a normal diet and free access to water. After animal pre-acclimatization, basal blood glucose and body weight were measured in mice before administration (Day 0). Subsequently, mice were administered subcutaneously (sc) the following drugs: a solvent control (Vehicle), semaglutide (10 nmol / kg, QD), cagrintide (10 nmol / kg, Q2D), AMY-015 (10 nmol / kg, Q2D), and a combination of semaglutide (10 nmol / kg, QD) and AMY-015 (10 nmol / kg, Q2D). Administration continued for 21 days. Daily body weight and total daily food intake were measured after administration. Changes in body weight (% of initial value) and total daily food intake were calculated and plotted. At the end of administration, blood and adipose tissue were dissected, adipose tissue was weighed, and blood biochemical indicators were measured. The results show that the compounds of this invention have a superior weight-loss effect compared to Cagrilintide, and the combination with smegglutinin is more effective in reducing rat body weight and food intake, while also protecting muscle mass to some extent from loss. The results of AMY-015 are exemplarily shown in this test example; see [link to test results]. Figure 8 Test Example 6: Changes in body weight, blood glucose, and food intake in a ZDF rat model after continuous drug administration.

[0503] Adult male ZDF rats, weighing 350-400g, were used in the experiment and fed a diabetic model diet. These rats were randomly divided into groups of eight, and kept in individual cages for one week of acclimatization, during which they received a normal diet and free access to water. After the animal pre-acclimatization, basal blood glucose and body weight were measured before administration (Day 0). Subsequently, the rats were administered subcutaneously (sc) the following compounds: vehicle (solvent control), Cagrilintide (30 nmol / kg, Q2D), and AMY-015 (30 nmol / kg, Q2D), for 28 consecutive days. Blood glucose levels were measured every two days after administration, and the change in blood glucose (% of initial value) was calculated to plot a blood glucose-time curve. The results showed that the compound of the present invention could reduce blood glucose in rats, superior to Cagrilintide. The results of AMY-015 are exemplarily shown in this test example; see [link to relevant documentation]. Figure 9 .

[0504] Test Example 7: Thioflavin T (ThT) Fibrosis Experiment

[0505] Amylin analogues, due to their structural characteristics, are prone to fibrotic aggregation in vivo and in vitro, resulting in demanding storage conditions and potential immunogenic risks. This invention employs a helical fixation technique to stabilize the secondary structure of peptide chains, reduce the stacking of hydrophobic groups between chains, and effectively reduce the tendency for fibrosis. Prepare a PBS solution (1 mg / mL) of the test peptide and a PBS working solution (5 mM) of ThT. At room temperature, take 400 μL of the test peptide sample (the peptide derivatives prepared in Examples 1 and 2 (i.e., AMY-001 to AMY-041)) into a 1.5 mL EP tube, add 8 μL of ThT stock solution, vortex to mix, centrifuge at low speed to ensure no residue buildup, and transfer the liquid to a 96-well plate, 200 μL per well. Multiple sets can be set up in parallel. Seal the wells with 200 μL of PBS and attach a transparent membrane. Kinetic tests were performed using a Biotek microplate analyzer, with monitoring points spaced 15 minutes apart. The excitation wave was 440 nm, and the emission wave was 485 nm. ΔFold change was calculated as (detected value / average of the first ten monitoring points). The horizontal axis represents time (h), and the vertical axis represents ΔFold change. Graphpad nonlinear fitting was used to plot the results, revealing that the polypeptide derivatives of this invention significantly reduce fibrosis and aggregation, exhibiting higher stability. This embodiment exemplifies the results of some polypeptide derivatives; for details, please refer to [link to documentation]. Figure 10 .

[0506] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0507] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. A polypeptide derivative or a pharmaceutically acceptable salt thereof, characterized in that, The sequence of the polypeptide is at least one of the following: U-ASHLSTCVLGRLSCELHKL-Aib-DYPRTDVGAGSP-NH2; U-ASHLSTACLGRLSACLHKL-Aib-DYPRTDVGAGSP-NH2; U-ASHLSTAVLCRLSAELCKL-Aib-DYPRTDVGAGSP-NH2; U-ASHLSTAVLGCLSAELHCL-Aib-DYPRTDVGAGSP-NH2; U-ASHLSTAVLGRCSAELHKC-Aib-DYPRTDVGAGSP-NH2; U-ASHLSTAVLGRLCAELHKLCDYPRTDVGAGSP-NH2; U-ASHLSTACLGRLSACLH-Orn-L-Aib-D-(αMeF)-PRTDVGAGS-Hyp-NH2; U-ASH-(αMeL)-STACLRLSACLHKL-Aib-D-(αMeF)-P-(N-Me-R)-TDVGAGSP-NH2; U-ASHLSTACLGRLSACLH-(αMeK)-L-Aib-DYP-(N-Me-R)-TDVGAGS-Hyp-NH2; U-ASHLSTACLG-homoR-LSACLH-(αMeK)-L-Aib-DYP-(N-Me-R)-TDVGAGSP-NH2; U-ASHLSTACLGRLSACLH-(αMeK)-L-Aib-DYP-norR-TDVGAGS-Hyp-NH2; U-ASHLSTACLGRLSACLH-(αMeK)-L-Aib-DYPQTDVGAGS-Hyp-NH2; U-ASHLSTAKLGRLSAKLH-(αMeK)-L-Aib-DYPQTDVGAGS-Hyp-NH2; U-ASHLSTAVLGKLSAELHKL-Aib-DYPQTDVGAGS-Hyp-NH2; Each U group is independently selected from acetyl groups; The polypeptide derivative further includes a modifying group, which is linked to two amino acid sites in the sequence of the polypeptide, at positions X and X, respectively. i and X i+7 , where i is any integer between 6 and 13; The two amino acids linked to the modified group of the polypeptide are both C or K. When both amino acids are C, the modifying group is connected to the -SH of the side chain of the amino acid through a sulfur-carbon bond; When both amino acids are K, the modifying group is connected to the -NH2 of the side chain of the amino acid via an amide bond; The polypeptide derivative has any of the structures shown in Table A or Table B: Table A Table B The structure of the modifying group A1 is as follows: ； The structure of the modifying group A12 is as follows: ； The structure of the modifying group B1 is as follows: ； The structure of the modifying group B16 is as follows: 。 2. A pharmaceutical composition, characterized in that, This includes the polypeptide derivatives of claim 1 or their pharmaceutically acceptable salts.

3. The pharmaceutical composition according to claim 2, characterized in that, This further includes pharmaceutically acceptable excipients or carriers.

4. A combination drug or pillbox, characterized in that, include: The polypeptide derivative of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 2 or 3, as the first active ingredient; and the second active ingredient; The second active ingredient includes drugs for the prevention and / or treatment of diseases.

5. The combined drug or cassette according to claim 4, characterized in that, The diseases mentioned include diseases related to amylin receptor and / or calcitonin receptor.

6. The combined drug or cassette according to claim 5, characterized in that, The diseases related to the amylin receptor and / or calcitonin receptor include lipid metabolism disorders, glucose metabolism disorders, cardiovascular diseases, brain diseases, mental diseases, or nervous system diseases.

7. The combined drug or cassette according to claim 5, characterized in that, The amylin receptor and / or calcitonin receptor-related diseases include at least one of the following: metabolic disorder-related diseases, bone-related diseases, cardiovascular diseases not related to metabolic diseases, symptoms or diseases related to alcohol or drug addiction, and neurodegenerative diseases.

8. The combined drug or cassette according to claim 5, characterized in that, The amylin receptor and / or calcitonin receptor-related diseases include at least one of the following: diabetes mellitus, hypertension, arteriosclerosis, cirrhosis, coronary heart disease, angina pectoris, myocardial infarction, inflammatory bowel disease, dyspepsia, gastrointestinal ulcers, hyperglycemia, impaired glucose tolerance, syndrome X, cognitive impairment, stroke, dyslipidemia-related diseases, metabolic syndrome, abnormal weight, obesity, fatty liver disease, diabetic nephropathy, diabetes-associated renal fibrosis, liver fibrosis, Alzheimer's disease, and Parkinson's disease.

9. The combined drug or cassette according to claim 5, characterized in that, The diseases associated with the amylin receptor and / or calcitonin receptor include non-alcoholic steatohepatitis.

10. The combined drug or cassette according to claim 5, characterized in that, The diseases associated with the amylin receptor and / or calcitonin receptor include at least one of cardiovascular disease, diabetes, and / or obesity.

11. The combined drug or cassette according to claim 4, characterized in that, The drug is selected from at least one of the following: adrenergic receptor blockers, HMG-CoA reductase inhibitors, angiotensin receptor antagonists, angiotensin-converting enzyme inhibitors, calcium channel blockers, endothelin antagonists, renin inhibitors, diuretics, aldosterone receptor blockers, endothelin receptor blockers, aldosterone synthase inhibitors, CETP inhibitors, relaxin, PCSK9 inhibitors, BNP and NEP inhibitors, GLP-1 analogs, insulin, sulfonylureas, biguanides, megglitazones, glucosidase inhibitors, DPP IV inhibitors, and SGLT2 inhibitors.

12. The combined drug or cassette according to claim 4, characterized in that, The drug is selected from one of the following: GLP-1 analogs, selenoquine hypoglycemic agents, biguanides, and acarbose.

13. The combined drug or cassette according to claim 12, characterized in that, The GLP-1 analogues include at least one of semaglutide, exenatide, liraglutide, dulaglutide, telposide, and retaglutide.

14. The combined drug or flask according to claim 12, characterized in that, The levofloxacin class of hypoglycemic drugs includes at least one of dapagliflozin and empagliflozin.

15. The combined drug or cassette according to claim 12, characterized in that, The biguanides include metformin.

16. Use in the preparation of a medicament by the polypeptide derivative of claim 1 or a pharmaceutically acceptable salt thereof, the pharmaceutical composition of claim 2 or 3, or any combination medicament or medicament of claims 4-15, wherein the medicament is for the treatment or prevention of amylin receptor and / or calcitonin receptor-related diseases, wherein the amylin receptor and / or calcitonin receptor-related diseases are diabetes or obesity.

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