Human amylin analogs, derivatives thereof and uses thereof

Abstract

This application relates to polypeptide analogs, or derivatives thereof, and methods for their preparation and use, and particularly to polypeptide analogs or derivatives thereof having agonistic activity against the islet amyloid polypeptide receptor. The peptide analogs or derivatives thereof provided in this application have good agonistic activity against the islet amyloid polypeptide receptor, and at the same time have characteristics such as good stability and high activity, and can be used for the treatment and prevention of diseases related to the islet amyloid polypeptide receptor, such as abnormal body weight, particularly obesity and overweight.

Description

[Technical Field]

[0001] This application relates to the field of biomedical or biopharmaceutical technology, and more particularly to human amylin analogs, derivatives, and their use in the treatment and / or prevention of diseases related to amylin or metabolism. [Background technology]

[0002] Amylin, also known as islet amyloid polypeptide (IAPP), is a member of the calcitonin gene-related peptide (CGRP) family. It exerts physiological functions by activating its corresponding receptor, exerting diverse functions such as regulating blood glucose levels, food intake, and metabolism. However, amylin is prone to fibrillation and amyloid deposit formation under neutral or alkaline conditions. At the same time, amylin's short residence time in the blood and half-life of only approximately 13 minutes make it difficult to use, limiting its use in disease treatment. Therefore, the development of amylin analogs with better stability, longer half-lives, and more pronounced therapeutic effects is of great importance for the treatment of metabolic diseases such as diabetes and obesity.

[0003] Pramlintide (also known as Symlin) is an amylin analogue whose stability in solution is improved by substituting Pro for the 25th, 28th, and 29th amino acid residues. It is currently approved for the treatment of diabetes. However, pramlintide's half-life is only approximately 48 minutes, requiring frequent injections to maintain its efficacy. Its tendency to aggregate and precipitate under neutral pH conditions limits its use in combination with other therapeutic agents. WO2012162542A1 discloses derivatives of amylin polypeptide analogues that are substituted with lysine and conjugated with polyethylene glycol to extend the in vivo action of amylin or its analogues. However, the polyethylene glycolated derivatives have significantly reduced affinity for amylin receptors. Li-Mei Yan et al. (Proc Natl Acad Sci, 2006, 103(7):2046-2051) described an amylin derivative, IAPP-GI, in which the solubility of amylin was improved by methylation of 24-Gly and 26-Ile, but the activity of the amylin derivative was reduced compared to the wild-type amylin. Cagrilintide, a long-acting amylin derivative developed by Novo Nordisk, is currently in Phase II clinical trials.

[0004] Although the prior art amylin analogues / derivatives have been improved in terms of solubility, stability, half-life, activity, etc. compared with amylin, technical problems such as insufficient efficacy in the body and poor drug stability still exist, and therefore amylin derivatives require further improvement. Summary of the Invention

[0005] The present application provides novel amylin analogs and derivatives of the analogs, methods of preparation, pharmaceutical compositions comprising the polypeptides or derivatives thereof, products, and their use in the prevention and / or treatment of disease.

[0006] In one aspect, the present application provides a novel amylin analog or a pharmaceutically acceptable salt thereof. The analog is X1CNTA TCATQ RLAX 14 F LX 17 HX 19 X 20 X 21 X 22 X 23 X 24 X 25 ILX 28 X 29 T NVGSX 35 TX 37 The amino acid sequence comprises an amino acid sequence represented by formula (I), wherein the amino acid sequence comprises one or more amino acid mutations at positions 1, 14, 17, 19, 20, 21, 22, 23, 24, 25, 28, 29, 35, and 37. Preferably, X1 is selected from K, D, Dab, Dap, E, H, Har, Orn, R, or a deletion; 14 is selected from N, Aad, Dab, Dap, E or D, and X 17 is selected from V, Orn, or R, and X 19 is selected from S or K, and X 20 is selected from S or K, and X 21 is selected from N or K, and X 22 is selected from N or K, and X 23 is selected from F, L, or K, and X 24 is selected from G, R, H, P, D, N, S, K, αK, Dap, Dab, Orn, Cit, Sar, Trx or AOC, and X 25 is selected from A, K, or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 35 is selected from N or G, and X 37 is selected from Y, 3-Am-Pro, Aze, cis-Hyp, P, Pip, αMePro, or trans-Hyp.

[0007] In some preferred embodiments, the amylin analog is X1CNTA TCATQ RLAX 14 F LX 17HSS NNX 23 X 24 X 25 ILX 28 X 29 T NVGSN TX 37 The amino acid sequence comprises an amino acid sequence represented by formula (II), which comprises one or more amino acid mutations at positions 1, 14, 17, 23, 24, 25, 28, 29, and 37.

[0008] Preferably, X1 is selected from K, D, Dab, Dap, E, H, Har, Orn, R or deletion; 14 is selected from N, Aad, Dab, Dap, E or D, and X 17 is selected from V, Orn or R, X 23 is selected from F, L or K, and X 24 is selected from G, R, H, P, D, N, S, K, αK, Dap, Dab, Orn, Cit, Sar, Trx or AOC, and X 25 is selected from A, K or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is selected from Y, 3-Am-Pro, Aze, cis-Hyp, P, Pip, αMePro, or trans-Hyp.

[0009] Preferably, X1 is selected from K or R, and X 14 is selected from N, Aad, Dab, Dap, E or D, and X 17 is selected from V, Orn or R, X 23 is selected from F, L or K, and X 24 is selected from G, R, H, P, D, N, S, K, αK, Dap, Dab, Orn, Cit, Sar, Trx or AOC, and X 25 is selected from A, K or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is selected from Y, 3-Am-Pro, Aze, cis-Hyp, P, Pip, αMePro, or trans-Hyp.

[0010] Preferably, X1 is selected from K, D, Dab, Dap, E, H, Har, Orn, R or deletion; 14 is selected from N, Aad, Dab, Dap, E or D, and X 17 is selected from V, Orn or R, X 23 is selected from F, L or K, and X 24 is selected from G, R or K, and X 25 is selected from A, K or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is selected from Y, 3-Am-Pro, Aze, cis-Hyp, P, Pip, αMePro, or trans-Hyp.

[0011] Preferably, X1 is selected from K or R, and X 14 is selected from N, Aad, Dab, Dap, E or D, and X 17 is selected from V, Orn or R, X 23 is selected from F, L or K, and X 24 is selected from G, R or K, and X 25 is selected from A, K or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is selected from Y, 3-Am-Pro, Aze, cis-Hyp, P, Pip, αMePro, or trans-Hyp.

[0012] Preferably, X1 is selected from K, D, Dab, Dap, E, H, Har, Orn, R or deletion; 14 is selected from N, Aad, Dab, Dap, E or D, and X 17 is selected from V, Orn or R, X 23 is selected from F, L or K, and X 24 is selected from G, R, H, P, D, N, S, K, αK, Dap, Dab, Orn, Cit, Sar, Trx or AOC, and X 25 is selected from A, K or P, and X28 is selected from S or P, and X 29 is selected from S or P, and X 37 is P or trans-Hyp.

[0013] Preferably, X1 is selected from K or R, and X 14 is selected from N, Aad, Dab, Dap, E or D, and X 17 is selected from V, Orn or R, X 23 is selected from F, L or K, and X 24 is selected from G, R or K, and X 25 is selected from A, K or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is P or trans-Hyp.

[0014] Particularly preferably, X1 is selected from K or R, and X 14 is selected from N or D, and X 17 is selected from V or R, X 23 is selected from F or L, and X 24 is selected from G, R or K, and X 25 is selected from A or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is selected from Y, P or trans-Hyp.

[0015] More preferably, X1 is K and X 14 is selected from N or D, and X 17 is selected from V or R, and X 23 is selected from F or L, and X 24 is selected from G, R or K, and X 25 is selected from A or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is selected from Y, P or trans-Hyp.

[0016] More preferably, X is R and X 14 is selected from N or D, and X 17 is selected from V or R, and X 23 is selected from F or L, and X 24 is selected from G, R or K, and X 25 is selected from A or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is selected from Y, P or trans-Hyp.

[0017] More preferably, X1 is selected from K or R, and X 14 is D and X 17 is R and X 23 is L and X 25 is P and X 24 is selected from G, R or K, and X 28 is P and X 29 is P and X 37 is P or trans-Hyp.

[0018] More preferably, X1 is K and X 14 is D and X 17 is R and X 23 is L and X 24 is selected from G or K, and X 25 is P and X 28 is P and X 29 is P and X 37 is P or trans-Hyp.

[0019] Most preferably, X is K and X 14 is D and X 17 is R and X 23 is L and X 24 is K and X 25 is P and X 28 is P and X 29 is P and X 37 is P or trans-Hyp.

[0020] In one aspect, the present application provides novel amylin analog derivatives or pharmaceutically acceptable salts thereof, wherein the amylin analog derivatives have one or more amino acid modifications, and the analog derivatives include the polypeptide sequences of the amylin analogs described above. In some embodiments, the polypeptide analogs or derivatives thereof described herein comprise the amino acid sequences of SEQ ID NOs: 2-104 shown in Table 1.

[0021] [Table 1(1)] [Table 1(2)] [Table 1(3)]

[0022] In some embodiments, the derivative sequences of the polypeptide analogs described herein contain 1 to 10 amino acid modifications, preferably 1 to 8 amino acid modifications, even more preferably 1 to 4 amino acid modifications, more preferably 2 amino acid modifications, and most preferably 1 amino acid modification, compared to the wild-type polypeptide sequence.

[0023] In some embodiments, the amino acid modification of the derivative sequence of the polypeptide analog, compared to the wild-type polypeptide sequence, is the introduction of a functional group that can be used for coupling to the side chain of a natural or unnatural amino acid (including, but not limited to, glutamic acid, aspartic acid, lysine, glutamine, or asparagine). Preferably, the amino acid modification is on the side chain of a lysine, preferably the lysine is located at position 1 of the polypeptide derivative, and the modified side chain is selected from the alpha-amino side chain or the epsilon-amino side chain of a lysine, preferably the epsilon-amino side chain.

[0024] In some embodiments, the functional group that can be used for coupling includes an extension portion and a linker that connects the extension portion and the amino acid to be modified, where the linker may be composed of one or more linkers, and the order of connecting the linkers is arbitrary, and should be understood as within the scope of the present application as long as it can connect the extension portion and the amino acid to be modified.

[0025] In a preferred embodiment, the single linker may have the following structure: [ka] [ka] Here, m is 0, 1, 2, or 3; n is 1, 2, or 3; s is any integer from 0 to 6; and p is any integer from 1 to 8.

[0026] Preferably, the linker is: [ka] and Here, m is 1 or 2, n is 1 or 2, and p is any integer from 1 to 5.

[0027] More preferably, the linker is: [ka] where p is any integer from 1 to 5; Preferably, p is 1 and the linker is γGlu.

[0028] In a preferred embodiment, the extension portion comprises: [ka] where x is an integer between 4 and 38.

[0029] Preferably, the extension portion is [ka] where x is an integer between 4 and 38.

[0030] Preferably, the amino acid sequence of the polypeptide analog comprises the amino acid sequence of formula (I): X1CNTATCATQRLAX 14 FLX 17 HX 19 X 20 X 21 X 22 X 23 X 24 X 25 ILX 28 X 29 TNVGSX 35 TX 37 Formula (I), where X1 is selected from K or R, and X 14 is selected from N, Aad, Dab, Dap, E or D, and X 17 is selected from V, Orn or R, X 23 is selected from F, L or K, and X 24 is selected from G, R, H, P, D, N, S, K, αK, Dap, Dab, Orn, Cit, Sar, Trx or AOC, and X 25 is selected from A, K or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is selected from Y, 3-Am-Pro, Aze, cis-Hyp, P, Pip, αMePro or trans-Hyp. [ka] and Here, m is 1 or 2, n is 1 or 2, and p is an integer of 1 to 5. [ka] where x is an integer between 4 and 38.

[0031] Preferably, the amino acid sequence of the polypeptide analog comprises the amino acid sequence of formula (I): X1CNTATCATQRLAX 14 FLX 17 HX 19 X 20 X 21 X 22 X 23 X 24 X 25 ILX 28 X 29 TNVGSX 35 TX 37 Formula (I), wherein X1 is selected from K, D, Dab, Dap, E, H, Har, Orn, R or deletion; 14 is selected from N, Aad, Dab, Dap, E or D, and X 17 is selected from V, Orn or R, X 23 is selected from F, L or K, and X 24 is selected from G, R or K, and X 25 is selected from A, K or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is selected from Y, 3-Am-Pro, Aze, cis-Hyp, P, Pip, αMePro or trans-Hyp. [ka] and Here, m is 1 or 2, n is 1 or 2, and p is an integer of 1 to 5. [ka] where x is an integer between 4 and 38.

[0032] Preferably, the amino acid sequence of the polypeptide analog comprises the amino acid sequence of formula (I): X1CNTATCATQRLAX 14 FLX 17 HX19 X 20 X 21 X 22 X 23 X 24 X 25 ILX 28 X 29 TNVGSX 35 TX 37 Formula (I), where X1 is selected from K or R, and X 14 is selected from N, Aad, Dab, Dap, E or D, and X 17 is selected from V, Orn or R, X 23 is selected from F, L or K, and X 24 is selected from G, R or K, and X 25 is selected from A, K or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is selected from Y, 3-Am-Pro, Aze, cis-Hyp, P, Pip, αMePro or trans-Hyp. Particularly preferably, X is selected from K or R and X 14 is selected from N or D, and X 17 is selected from V or R, and X 23 is selected from F or L, and X 24 is selected from G or K, and X 25 is selected from A or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is selected from Y or trans-Hyp. [ka] and Here, m is 1 or 2, n is 1 or 2, and p is an integer of 1 to 5. [ka] where x is an integer between 4 and 38.

[0033] Preferably, the amino acid sequence of the polypeptide analog comprises the amino acid sequence of formula (I): X1CNTATCATQRLAX 14 FLX 17 HX 19 X 20 X 21 X 22 X 23 X 24 X 25 ILX 28 X 29 TNVGSX 35 TX 37 Formula (I), where X1 is K and X 14 is selected from N or D, and X 17 is selected from V or R, and X 23 is selected from F or L, and X 24 is selected from G or K, and X 25 is selected from A or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is selected from Y or trans-Hyp. [ka] and Here, m is 1 or 2, n is 1 or 2, and p is an integer of 1 to 5. [ka] where x is an integer between 4 and 38.

[0034] Preferably, the amino acid sequence of the polypeptide analog comprises the amino acid sequence of formula (I): X1CNTATCATQRLAX 14 FLX 17 HX 19 X 20 X 21 X 22 X 23 X 24 X 25 ILX28 X 29 TNVGSX 35 TX 37 Formula (I), where X1 is R and X 14 is selected from N or D, and X 17 is selected from V or R, and X 23 is selected from F or L, and X 24 is selected from G or K, and X 25 is selected from A or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is selected from Y or trans-Hyp. [ka] and Here, m is 1 or 2, n is 1 or 2, and p is an integer of 1 to 5. [ka] where x is an integer between 4 and 38.

[0035] Preferably, the amino acid sequence of the polypeptide analog comprises the amino acid sequence of formula (I): X1CNTATCATQRLAX 14 FLX 17 HX 19 X 20 X 21 X 22 X 23 X 24 X 25 ILX 28 X 29 TNVGSX 35 TX 37 Formula (I), where X1 is selected from K or R, and X 14 is D and X 17 is R and X 23 is L and X 25 is P and X 24 is selected from G or K, and X28 is P and X 29 is P and X 37 is trans-Hyp. The linker is: [ka] and Here, m is 1 or 2, n is 1 or 2, and p is an integer of 1 to 5. [ka] where x is an integer between 4 and 38.

[0036] Preferably, the amino acid sequence of the polypeptide analog comprises the amino acid sequence of formula (I): X1CNTATCATQRLAX 14 FLX 17 HX 19 X 20 X 21 X 22 X 23 X 24 X 25 ILX 28 X 29 TNVGSX 35 TX 37 Formula (I), where X1 is K and X 14 is D and X 17 is R and X 23 is L and X 24 is selected from G or K, and X 25 is P and X 28 is P and X 29 is P and X 37 is trans-Hyp. The linker is: [ka] and Here, m is 1 or 2, n is 1 or 2, and p is an integer of 1 to 5. [ka] where x is an integer between 4 and 38.

[0037] Preferably, the amino acid sequence of the polypeptide analog comprises the amino acid sequence of formula (I): X1CNTATCATQRLAX 14 FLX 17 HX 19 X 20 X 21 X 22 X 23 X 24 X 25 ILX 28 X 29 TNVGSX 35 TX 37 Formula (I), where X1 is K and X 14 is D and X 17 is R and X 23 is L and X 24 is K and X 25 is P and X 28 is P and X 29 is P and X 37 is trans-Hyp. The linker is: [ka] and Here, m is 1 or 2, n is 1 or 2, and p is an integer of 1 to 5. [ka] where x is an integer between 4 and 38.

[0038] Preferably, the amino acid sequence of the polypeptide analog comprises the amino acid sequence of formula (I): X1CNTATCATQRLAX 14 FLX 17 HX 19 X 20 X 21 X 22 X 23 X 24 X25 ILX 28 X 29 TNVGSX 35 TX 37 Formula (I), X1 is selected from K, D, Dab, Dap, E, H, Har, Orn, R or deletion; 14 is selected from N, Aad, Dab, Dap, E or D, and X 17 is selected from V, Orn or R, X 23 is selected from F, L or K, and X 24 is selected from G, R, H, P, D, N, S, K, αK, Dap, Dab, Orn, Cit, Sar, Trx or AOC, and X 25 is selected from A, K or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is P or trans-Hyp. The linker is: [ka] and Here, m is 1 or 2, n is 1 or 2, and p is an integer of 1 to 5. [ka] where x is an integer between 4 and 38.

[0039] Preferably, the amino acid sequence of the polypeptide analog comprises the amino acid sequence of formula (I): X1CNTATCATQRLAX 14 FLX 17 HX 19 X 20 X 21 X 22 X 23 X 24 X 25 ILX 28 X 29 TNVGSX 35 TX 37 Formula (I), X1 is selected from K or R, and X 14is selected from N, Aad, Dab, Dap, E or D, and X 17 is selected from V, Orn or R, X 23 is selected from F, L or K, and X 24 is selected from G, R or K, and X 25 is selected from A, K or P, and X 28 is selected from S or P, and X 29 is selected from S or P, and X 37 is P or trans-Hyp. The linker is: [ka] and Here, m is 1 or 2, n is 1 or 2, and p is an integer of 1 to 5. [ka] where x is an integer between 4 and 38.

[0040] In some embodiments, a derivative of a polypeptide analog of the present application is selected from the following compounds shown in Table 2, where PD065 is a Cagrilintide molecule.

[0041] [Table 2(1)] [Table 2(2)] [Table 2(3)] [Table 2(4)] [Table 2(5)] [Table 2(6)] [Table 2(7)] [Table 2(8)] [Table 2(9)]

[0042] Compounds PD001 to PD198 represent polypeptide derivatives obtained by modifying an amino acid sequence with a fatty acid-containing side chain. The term "fatty acid-containing side chain" refers to a functional group that can be used for coupling and is introduced into the side chain of an amino acid to modify the amino acid. Examples include derivatives obtained by modifying a corresponding polypeptide sequence with a corresponding fatty acid-containing side chain in Table 2. For example, PD001 represents a derivative obtained by modifying the 21st amino acid, K, of the sequence RCNTATCATQRLAEFLRHSSKNLGPILPPTNVGSNTP-NH2 with a functional group that can be used for coupling, i.e., the side chain (2*AEEA+γGlu+C20 diacid). Typically, the C16-C22 lipid diacid moiety is located outside the fatty acid-containing side chain used for modification of the amino acid sequence, distal to the connection between the side chain and the amino acid sequence.

[0043] In some embodiments, derivatives of the polypeptide analogs of the present application are preferably selected from the following derivatives: PD069, PD070, PD071, PD072, PD140, PD158, PD160, PD161, PD144, PD145, PD146, PD147, PD187.

[0044] In some embodiments, the polypeptide analog derivative of the present application is the compound PD069, the structure of which is as follows: [ka]

[0045] In some embodiments, the polypeptide analog derivative of the present application is the compound PD070, the structure of which is as follows: [ka]

[0046] In some embodiments, the polypeptide analog derivative of the present application is the compound PD071, the structure of which is as follows: [ka]

[0047] In some embodiments, the polypeptide analog derivative of the present application is the compound PD072, the structure of which is as follows: [ka]

[0048] One aspect of the present application also relates to a method for preparing said novel amylin analogs or derivatives thereof.

[0049] In some embodiments, the novel amylin analog or its derivative can be prepared by conventional methods known to those skilled in the art. Typically, polypeptide molecules can be prepared by chemical synthesis, biological fermentation, or a combination of both. For example, during the polypeptide synthesis process, a polypeptide gene sequence is synthesized, and the corresponding polypeptide gene with a tag is inserted into a plasmid using conventional genetic engineering methods to construct an expression plasmid. The expression plasmid is then introduced into competent cells to construct expression cells. The expression cells containing the target polypeptide gene after screening are fermented under appropriate culture conditions, the fermentation product is recovered, and the target polypeptide can finally be obtained by purification. Furthermore, a polypeptide analog derivative can be obtained by chemically modifying the amino acid side chain of the target polypeptide and purifying it.

[0050] In one aspect, the present application also relates to medicaments or pharmaceutical compositions comprising the amylin analogs or derivatives thereof, and pharmaceutically acceptable salts of the amylin analogs or derivatives thereof.

[0051] In some embodiments, the pharmaceutical compositions described herein further comprise one or more oral delivery agents. The oral delivery agent is preferably N-[8-(2-hydroxybenzoyl)amino]octanoic acid (NAC) salt, more preferably PNAC, i.e., potassium N-[8-(2-hydroxybenzoyl)amino]octanoate. The NAC salt or PNAC salt of the oral delivery agent described herein should be understood to be any crystalline form that can satisfy the oral delivery form of the composition of the present application, as described in CN112661663B.

[0052] The structure of the PNAC salt is represented by formula (II): [ka]

[0053] The structure of the NAC salt is represented by formula (III): [ka]

[0054] In some embodiments, the pharmaceutical compositions described herein further comprise one or more other pharmacologically active substances.

[0055] In some embodiments, the compositions described herein further comprise a pharmaceutically acceptable auxiliary substance. Preferably, the pharmaceutically acceptable auxiliary substance includes, but is not limited to, one or more of the following: a pharmaceutically acceptable carrier, a stabilizer, a dispersant, a cosolvent, a plasticizer, etc. Typically, these are non-toxic, inert, and pharmaceutically acceptable inert substances.

[0056] In one aspect, the present application provides use of an amylin analog or derivative thereof, or pharmaceutical composition described herein in the preparation of a medicament for preventing and / or treating a disease. Preferably, the use includes use in the preparation of a medicament for preventing and / or treating an amylin receptor-related disease. More preferably, the use includes use in the preparation of a medicament for preventing and / or treating a lipid metabolism disorder, a blood glucose metabolism disorder, a cardiovascular disease, a cerebrovascular disease, a psychiatric disease, or a nervous system disease, including but not limited to, body weight disorder, hyperlipidemia, hypertension, arteriosclerosis, fatty liver, dyslipidemia, cirrhosis, coronary heart disease, angina pectoris, cardiac infarction, inflammatory bowel disease, dyspepsia and gastrointestinal ulcer, hyperglycemia, diabetes, impaired glucose tolerance, syndrome X, cognitive impairment, stroke, etc. Preferably, the disease is a body weight disorder or a blood glucose metabolism disorder, more preferably, the disease is a body weight disorder, including but not limited to, obesity or overweight.

[0057] In one aspect, the present application provides a method for preventing and / or treating a disease. The method comprises administering to a subject in need thereof an amylin analog or derivative thereof, drug, or pharmaceutical composition described herein. The disease is an amylin receptor-related disease, preferably a lipid metabolism disorder, a blood glucose metabolism disorder, a cardiovascular disease, a cerebrovascular disease, a psychiatric disease, or a nervous system disease, including, but not limited to, weight disorder, hyperlipidemia, hypertension, arteriosclerosis, fatty liver, dyslipidemia, cirrhosis, coronary heart disease, angina pectoris, cardiac infarction, inflammatory bowel disease, dyspepsia and gastrointestinal ulcer, hyperglycemia, diabetes, impaired glucose tolerance, syndrome X, cognitive impairment, stroke, etc. More preferably, the disease is a weight disorder or a blood glucose metabolism disorder, and more preferably, the disease is a weight disorder, including, but not limited to, obesity or overweight.

[0058] The amylin analogs or derivatives thereof, drugs, or pharmaceutical compositions described herein can also be administered in combination with one or more other pharmacologically active substances, including, but not limited to, weight control agents, anti-obesity agents, lipid metabolism regulators, blood glucose regulators, antihypertensive agents, cardiovascular regulators, brain disease regulators, psychiatric disease regulators, or nervous system regulators, and preferably, the pharmacologically active substance is selected from GLP-1 receptor agonists (including, but not limited to, GLP-1, GLP-1 analogs, and GLP-1 derivatives), GIP receptor agonists (including, but not limited to, GIP, GIP analogs, and GIP derivatives), and insulin receptor agonists (including, but not limited to, insulin, insulin analogs, and insulin derivatives). [Effects of the Invention]

[0059] The polypeptide analogs or derivatives thereof provided in the present application have good agonistic activity against islet amyloid polypeptide receptors, and at the same time have characteristics such as good stability and high activity, and can be used for the treatment and prevention of diseases associated with islet amyloid polypeptide receptors, such as body weight disorders, particularly obesity and overweight. [Brief explanation of the drawings]

[0060] The accompanying drawings are intended to be used for a better understanding of the present application and are not to be construed as an undue limitation of the present application. [Figure 1] 1 shows the results of stability experiments at 40° C. on days 0, 7, and 15 for compounds PD065, PD069, PD070, PD071, and PD072, respectively. [Figure 2A] 1 shows the time-dependent change trends of the weight loss effects of different derivatives in SD rats. [Figure 2B] 1 shows the time-dependent change trends of the effects of different derivatives on food intake in SD rats. [Figure 2C] The weight change rate at 48 hours in SD rats treated with different derivatives in Group A is shown. [Figure 2D] The percentage change in body weight at 48 hours in SD rats treated with different derivatives in Group B is shown. [Figure 3A] 1 shows the time course change trends of the weight loss effects of compounds PD065 and PD069 at different concentrations in SD rats. [Figure 3B] 1 shows the time course change trends of the weight loss effects of compounds PD065 and PD070 at different concentrations in SD rats. [Figure 3C] 1 shows the time course change trends of the weight loss effects of compounds PD065 and PD071 at different concentrations in SD rats. [Figure 3D] 1 shows the time course change trends of the weight loss effects of compounds PD065 and PD072 at different concentrations in SD rats. [Figure 4A] 1 shows the time course trends of the effects of compounds PD065 and PD071 at different concentrations on food intake in SD rats. [Figure 4B] 1 shows the time course of changes in the effects of each derivative treatment group at different concentrations on food intake in SD rats. [Figure 5A] 1 shows the time-dependent change in the weight loss effect of compounds PD065 and PD187 in SD rats. [Figure 5B] 1 shows the time course of changes in the weight loss effect of the corresponding compounds in SD rats. [Figure 5C] 1 shows the time course of the effects of the corresponding compounds on food intake in SD rats. [Figure 6] 1 shows the pharmacokinetics of each polypeptide after oral administration of oral delivery compositions containing different polypeptide derivatives in beagle dogs. DETAILED DESCRIPTION OF THE INVENTION

[0061]

[0033] The following description of exemplary embodiments of the present application includes various details of the embodiments of the present application for ease of understanding, but these should be considered as mere examples. Therefore, it should be understood that those skilled in the art can make various changes and modifications to the embodiments described herein without departing from the scope and spirit of the present application. Also, for clarity and conciseness, the following description will omit descriptions of well-known functions and configurations.

[0062] Abbreviation The corresponding names or structural forms of some abbreviations used in the examples of this application are given in Table 3 below.

[0063] [Table 3]

[0064] AA: Amino Acid; Boc: t-Butyloxy carbonyl; DCM: dichloromethane; DMF: N,N-Dimethyl formamide; DIEA: N,N-Diisopropylethylamine; EDT: 1,2-Ethanedithiol; Fmoc: 9-fluorenylmethyloxycarbonyl; OtBu: tert-butyl ester; Pbf: 2,2,4,6,7-Pentamethyldihydrobenzofuran-5-sulfonyl chloride; Pip: Piperidine; TBTU: O-(Benzotriazol-l-yl)-N,N,N,N,-tetramethyluronium Tetrafluoroborate, O-(benzotriazol-1-yl)-N,N,N,,N,-tetramethyluronium tetrafluoroborate; tBu: tertiary butyl; TFA: Trifluoroacetic acid; TIS: Triisopropylsilane; Trt: Triphenylmethyl.

[0065] definition "Amylin" or "Amylin polypeptide" or "amylin" or "islet amyloid polypeptide" as used herein means the amino acid sequence Lys-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Asn-Phe-Leu-Val-His-Ser-Ser-Asn-Asn-Phe-Gly-Ala-Ile-Leu-Ser-Ser-Thr-Asn-Val-Gly-Ser-Asn-Thr-Tyr or a variant / analogue, fragment or fusion protein of said sequence which substantially retains an agonistic effect on the amylin receptor.

[0066] As used herein, "analog" refers to a compound formed by substituting at least one amino acid residue with another amino acid residue, and / or deleting at least one amino acid residue, and / or adding at least one amino acid residue to the N-terminus or C-terminus, and / or inserting at least one amino acid residue between any two amino acids in a particular polypeptide molecule.

[0067] In this application, unless the context clearly dictates otherwise, specific polypeptide names such as "amylin" and the terms "polypeptide," "analog," and "variant" may be substituted for each other.

[0068] As used herein, the term "derivative" refers to a product obtained by modifying a functional group (such as an amino acid residue) of a biological macromolecule (such as a polypeptide or protein) with a specific compound or molecule, including, but not limited to, acylation, amidation, esterification, and thioesterification.

[0069] The pharmaceutical compositions of the present application may be administered by any suitable route, such as orally, intranasally, intradermally, subcutaneously, intramuscularly or intravenously.

[0070] The term "lipid metabolism disorder"-related diseases described in this application is a general term for diseases caused by abnormalities in lipid metabolism, including not only general weight disorders (e.g., obesity) but also, for example, hyperlipidemia, hypertension, atherosclerosis, fatty liver, cirrhosis, coronary heart disease, angina pectoris, cardiac infarction, inflammatory bowel disease, dyspepsia, and gastrointestinal ulcers.

[0071] As used herein, "weight abnormality" refers to a subject's weight assessment index being outside the normal average range for that population, as assessed by a single assessment system. In one embodiment, the weight abnormality is obesity. In one embodiment, "weight loss" refers to a reduction in overweight or obesity.

[0072] Example The present application can be better understood with reference to the following examples. However, it should be understood that the following examples are for illustrative purposes only and should not be construed as limiting the scope of the present application in any way. Unless otherwise specified, the present application may be carried out in accordance with methods described in laboratory manuals such as "Molecular Cloning Laboratory Guide" and "Cell Laboratory Guide," which are well known to those skilled in the art, as well as CFDA experimental guidelines. All reagents and raw materials used herein are commercially available and can be purchased through public channels.

[0073] Example 1 Preparation of amylin derivatives Using solid-phase organic synthesis, the Fmoc-protected amino acid strategy and SPPS solid-phase synthesis technique were used to complete polypeptide synthesis, cleavage, oxidation, and purification to obtain the desired product.

[0074] Take the compound PD065 (Cagrilintide) as an example, its structural formula is as follows: [ka]

[0075] The synthesis process was as follows: Solid-phase synthesis: Using Fmoc-Linker MBHA resin with S = 0.32 mmol / g, Fmoc / tBu technology was employed. Amino acids were condensed and linked in order from the C-terminus to the N-terminus (from right to left) according to the sequence shown in Table 4.

[0076] [Table 4]

[0077] The amino acid coupling order is as follows: A-01Fmoc-Pro-OH,A-02Fmoc-Thr(tBu)-OH,A-03Fmoc-Asn(Trt)-OH,A-04Fmoc-Ser(tBu)-OH,A-05Fmoc-Gl y-OH,A-06Fmoc-Val-OH,A-07Fmoc-Asn(Trt)-OH,A-08Fmoc-Thr(tBu)-OH,A-09Fmoc-Pro-OH,A-10Fmoc-Pro -OH,A-11Fmoc-Leu-OH,A-12Fmoc-Ile-OH,A-13Fmoc-Pro-OH,A-14Fmoc-Gly-OH,A-15Fmoc-Phe-OH,A-16Fmo c-Asn(Trt)-OH,A-17Fmoc-Asn(Trt)-OH,A-18Fmoc-Ser(tBu)-OH,A-19Fmoc-Ser(tBu)-OH,A-20Fmoc-His(T rt)-OH,A-21Fmoc-Arg(Pbf)-OH,A-22Fmoc-Leu-OH,A-23Fmoc-Phe-OH,A-24Fmoc-Glu(OtBu)-OH,A-25Fmoc -Ala-OH,A-26Fmoc-Leu-OH,A-27Fmoc-Arg(Pbf)-OH,A-28Fmoc-Gln(Trt)-OH,A-29Fmoc-Thr(tBu)-OH,A-30 Fmoc-Ala-OH, A-31 Fmoc-Cys(Trt)-OH, A-32 Fmoc-Thr(tBu)-OH, A-33 Fmoc-Ala-OH, A-34 Fmoc-Thr(tBu)-OH, A-35 Fmoc-Asn(Trt)-OH, A-36 Fmoc-Cys(Trt)-OH, A-37 Fmoc-Lys(Boc)-OH, A-38 Fmoc-Glu-otbu, A-39 C20 diacid. Finally, the polypeptide derivative resin was formed. The polypeptide derivative resin was washed, transferred, and dried to a constant weight and awaited cleavage.

[0078] Cleavage of peptide resin: Preparation of cleavage reagent: Calculate the amount of cleavage reagent based on a ratio of 10 ml ± 2 ml of cleavage reagent to 1 g of peptide resin, with a TFA:HO:EDT:TIS ratio of 95:1:2:2. The required cleavage reagent, HO, TFA, EDT, and TIS, were added sequentially to the cleavage reaction bottle, with the temperature of the cleavage reagent controlled at 0-10°C. The cleavage reagent was added to the peptide resin with stirring. After the temperature of the reaction system stabilized, the temperature was controlled at 25-30°C and the reaction was stirred for 2.5 hours. The cleavage reagent was filtered off and precipitated with 5 times the volume of ice-cold diethyl ether. The precipitate was filtered, washed three times with 3 times the volume of ice-cold diethyl ether, and then dried under vacuum at room temperature to obtain a solid crude product.

[0079] Oxidation of polypeptide: The crude product was pulverized, purified water was prepared, and the pulverized crude product was slowly added under stirring. At the same time, aqueous acetonitrile solution was added dropwise, and the crude product was added and completely dissolved. After that, methyl iodoethanol solution was added and stirred for 30 minutes.

[0080] Purification and freeze-drying: The above oxidized liquid was filtered through a 0.45 μm microporous filter membrane, and the crude product was purified using a column prepared with a C-18 column filler. Separation and purification were performed using an appropriate gradient at room temperature, and the target product was collected, analyzed, tested, and classified. Purity of 90% or higher was required. The unqualified target product was collected and re-separated and purified using an appropriate gradient, and the qualified liquid peak was collected. The qualified liquid sample was freeze-dried under reduced pressure to obtain a freeze-dried powder of the purified polypeptide.

[0081] The compounds listed in Table 2 were prepared using similar methods and steps as described for compound PD065 (Cagrilintide).

[0082] Example 2 In vitro efficacy testing of amylin derivatives The purpose of this experiment was to detect the activity or potency of amylin derivatives on human amylin receptors in vitro using a luciferase assay.

[0083] 2.1 Construction of amylin receptor / CRE-luc cell line CHO-K1 / Ga15 / AMY3 cells (in which the calcitonin receptor and receptor-activating modified peptide RAMP had been constructed and purchased from GenScript) were transfected with a plasmid containing a multicopy cAMP response element (CRE)-driven luciferase expression frame by standard methods and cultured in F12 medium containing 200 μg / mL Zeocin, 2 μg / mL Puromycin, 100 μg / mL Hygromycin, and 400 μg / mL G418 to obtain a stably transfected amylin receptor / CRE-luc cell line.

[0084] 2.2 Amylin luciferase assay For activity measurements, 50 μL of growth medium (F12 medium containing 10% FBS) was added to each well of a white 96-well plate, and stably transfected amylin / CRE-luc CHO cells were seeded into the white plate at a density of approximately 20,000 cells / well.

[0085] The freeze-dried powder obtained in Example 1 was dissolved in 20 mM phosphate buffer, pH 7.0, and diluted with F12 medium containing 10% FBS to obtain a derivative sample with an initial concentration of 100 nM. -5 The derivative samples were gradient diluted with F12 medium containing 10% FBS to obtain seven samples with 10-fold concentration differences in the nM range. 50 μL of the corresponding sample solution was added to each well and incubated at 37°C and 5% CO2 for 24 hours. 100 μL of luciferase was then added to each well and incubated for 3 minutes. Finally, luminescence was measured using a SpectraMax L (Molecular Devices) with SoftMax Pro 7.0.3 GxP software. A standard curve was constructed based on the fluorescence values, and the EC50 was calculated. The results are shown in Table 5.

[0086] [Table 5]

[0087] The results in Table 5 demonstrate that all of the polypeptide derivatives of the present invention have good activity against human amylin receptors.

[0088] Example 3 Freeze-thaw stability of derivatives Lyophilized powder of the polypeptide analog derivative prepared in Example 1 at a mass-to-volume ratio of 0.2 mg / ml was mixed with buffer and the polypeptide was dissolved in 20 mM (pH 7.0) phosphate buffer (PB). After storing overnight at -20°C, the sample was removed and allowed to return to room temperature before being thawed again. During the experiment, changes in the properties of the polypeptide after dissolution and after re-freezing and thawing were observed and recorded, and the polypeptide concentrations before and after freezing and thawing were detected using the following liquid chromatography method:

[0089] Materials and equipment: high-performance liquid chromatograph (Agilent 1200), ultrapure water (18.2 MΩ homemade ultrapure water), acetonitrile (HPLC grade), trifluoroacetic acid (HPLC grade); column for reversed-phase chromatography: Sepax Bio-C4 4.6*100 mm 5 μm 300 Å.

[0090] Sample preparation: A fixed amount of polypeptide was weighed and dissolved in 20 mM PB buffer (pH 7.4). If solubility was poor, a small amount of 0.1 M NaOH was added to adjust the pH to 8.0 until complete dissolution. The concentration was controlled between 0.5 and 2 mg / ml. The sample to be measured was collected and centrifuged at 10,000 rpm at 4°C for 3 minutes, and the supernatant was transferred to a liquid injection bottle.

[0091] Chromatographic conditions: flow rate 1.0 ml / min; autosampler temperature 15°C; column temperature 15°C; detection wavelength 280 nm; mobile phase A: 100% H2O + 0.05% TFA; mobile phase B: 100% CAN. The elution gradient is shown in the table below.

[0092] [Table 6]

[0093] The freeze-thaw yield was calculated by the following formula: freeze-thaw yield = polypeptide concentration after freeze-thaw / polypeptide concentration before freeze-thaw. The relevant results are shown in Table 7.

[0094] [Table 7(1)] [Table 7(2)]

[0095] As shown in Table 7, most of the polypeptide derivatives of the present application have good freeze-thaw stability and yield.

[0096] Example 4: Study on the stability of derivatives at 40°C Compounds PD065, PD069, PD070, PD071, and PD072 were mixed with sodium dihydrogen phosphate (1.42 mg / ml) at 1.34 mg / ml, dissolved in ultrapure water, and the pH was adjusted to approximately 7.4 with hydrochloric acid / sodium hydroxide. The solution was then filtered through a 0.22 μm sterile filter in an ultraclean bench and placed into a sterile vial. The vial was capped and placed in a stability test chamber at 40°C.

[0097] Before the test, the sample was centrifuged at 10,000 rpm at 4°C for 3 minutes, and the supernatant was transferred to a liquid-phase injection bottle. The liquid-phase method described in Example 3 was used for detection. The results are shown in Table 8 and Figure 1.

[0098] [Table 8]

[0099] As shown in the results in Table 8, the thermal stability of the polypeptide derivatives of the present invention was almost the same as that of PD065, and unexpectedly, compound PD070 showed an extremely low rate of decrease in protein concentration.

[0100] Example 5: Study on the weight loss effect of derivatives Sprague Dawley rats weighing 200–250 g were selected for this experimental study. The rats arrived at least 10–14 days before the start of the experiment to allow them to adapt to the experimental environment. To ensure accurate data and high test sensitivity, the rats were housed under reversed light-dark conditions (i.e., lights off during the day and on at night) for two weeks after arrival and housed individually for the first week after arrival. Throughout the adaptation and experimental periods, rats had free access to food and water. The derivative test was divided into experimental groups A: A1, A2, A3, A4, A5, A6, and group B: B1, B2, B3, B4, B5. Each derivative treatment group consisted of five to eight rats. The rats in each group were subcutaneously administered the derivative or vehicle at a dose of 30 nmol / kg. The administration time for each group was recorded. After administration, the rats were returned to their home cages, where they were provided with food and water. Food consumption and changes in rat weight were recorded every 24 hours using online or manual recordings.

[0101] The changes in body weight and food intake of SD rats in each group are shown in Figures 2A to 2D and Tables 9 to 10.

[0102] [Table 9]

[0103] [Table 10]

[0104] The experimental results show that compared with the weight-reducing effect of PD065, the polypeptide derivatives of the present application have a higher weight-reducing ability, and can even reach a weight-reducing effect more than twice that of PD065.

[0105] Example 6: Study on weight loss effect depending on the dosage of derivatives This experimental study used SD rats obtained in the same manner as in Example 5. The derivative test was divided into experimental groups C (C1-C13) and D (D1-D7), with each derivative test group consisting of 5-8 rats. According to the administration schedules in Tables 11 and 12, the derivative or vehicle was subcutaneously administered to rats in each group at doses of 10 nmol / kg, 20 nmol / kg, or 30 nmol / kg, respectively. The administration time for each group was recorded. After administration, the rats were returned to their home cages where they were provided with food and water. Food consumption and changes in rat weight were recorded online or manually every 24 hours.

[0106] The changes in body weight and cumulative food intake of SD rats in each group are shown in Figures 3A to 3D, 4A to 4B and Tables 11 to 12.

[0107] [Table 11]

[0108] [Table 12]

[0109] The experimental results show that the polypeptide derivative molecule of the present invention has a better weight loss effect than PD065 (Cagrilintide) under the same conditions.

[0110] Example 7 Studies on the fatty acid side chains of derivatives This experimental study used SD rats obtained using the same method as in Example 5. The derivatives were tested in experimental group E, which tested the weight-loss effects of compounds PD065 and PD187, and experimental group F, which tested compounds PD065, PD071, PD140, PD144, PD145, PD146, and PD147. Each derivative test group consisted of 5 to 8 rats. Rats in each group were subcutaneously administered the derivative or vehicle at a dose of 30 nmol / kg. The administration time for each group was recorded. After administration, the rats were returned to their home cages, where food and water were available. Changes in food consumption and rat weight were recorded every 24 hours either online or manually. The results for experimental group E are shown in Figure 5A, and the results for experimental group F are shown in Figures 5B and 5C.

[0111] The results of animal experiments showed that when the N-terminus was R (PD140), the activity was superior to that of PD065.

[0112] To further evaluate the role of fatty acid side chains on polypeptide activity, PD071 (SEQ ID NO: 3) was used as a polypeptide backbone. The results showed that when different fatty acid linkers were used, such as 2AEEA + C20 diacid (PD187), αAsp + C20 diacid (PD144), βAsp + C20 diacid (PD145), Trx + C20 diacid (PD146), and Inp + C20 diacid (PD147), the activity was superior to that of PD065.

[0113] Example 8: Preparation of polypeptide derivative tablets containing fatty acid side chains The polypeptide derivatives containing fatty acid side chains were prepared as oral tablets containing the same amount of oral delivery agent PNAC. The preparation method for PNAC was as follows.

[0114] N-[8-(2-hydroxybenzoyl)amino]octanoic acid (NAC) was prepared by following the method described in Example 1 of International Patent Application WO2008 / 028859. Isopropyl alcohol (22070.0 ml, 4.0 vol) was added to a 50 L reactor, stirring was started, and NAC (5518 g, 1.0 eq) was added. The reaction system was heated to 50°C, and the prepared 50% potassium hydroxide solution (1304.0 g, 1.0 eq) was added dropwise to the reaction system. After the addition, the reaction system became a clear, transparent yellow solution, which was then incubated at 50°C for 1 hour. The reaction solution was batch-concentrated at 40°C to obtain a pale orange-yellow crude product.

[0115] The combined crude product was added to isopropanol (19310.0 ml, 3.5 vol) and slurried for 1 hour. The reaction system was suction filtered, and the filter cake was washed with isopropyl alcohol (2760.0 ml, 0.5 vol). The filter cake was transferred to a vacuum drying oven, the pressure of the drying system was balanced with nitrogen, and dried at 60°C for 16 hours. It was then transferred back to the vacuum drying oven and dried at 100°C for 24 hours. After drying, a total of 4.52 kg of product was obtained, with a yield of 72.8%. The product was an off-white powdery solid, i.e., PNAC.

[0116] The contents of the main components of the polypeptide derivative tablets containing fatty acid side chains are shown in Table 13.

[0117] [Table 13]

[0118] The preparation method of the polypeptide derivative tablets containing fatty acid side chains was as follows: the polypeptide derivative and PNAC were sieved, uniformly mixed with auxiliary ingredients, and directly compressed into tablets.

[0119] Example 9: Study on the Pharmacokinetic (PK) Properties of Polypeptide Derivative Tablets Containing Fatty Acid Side Chains The oral tablets prepared in Example 4 were orally administered once daily for five consecutive days to male beagle dogs (9-12 kg) aged 10-15 months. The administered dose of each fatty acid side chain-containing polypeptide derivative (i.e., PD065, PD187, PD183) was 7 mg / dog (n=5). The day of the first oral administration was recorded as day 1, and the day of the last oral administration was recorded as day 5. On day 1, whole blood was collected from the animals before administration (-10 min) and 2, 4, and 8 hours after administration, and plasma was prepared (heparin sodium was used as an anticoagulant). On days 2-4, whole blood was collected before administration (-10 min) and 2 and 4 hours after administration, and plasma was prepared (heparin sodium was used as an anticoagulant). On the fifth day of administration, plasma samples were prepared (-10 min) before administration and 2, 4, 8, 24, 48, and 72 hours after administration (heparin sodium was used as an anticoagulant). The plasma samples were stored at -80°C and used for subsequent analysis. The drug content in plasma was analyzed using LC-MS / MS (Waters ACQUITY I Class Premier UPLC tandem with Sciex 6500+QQQ). Experimental data were plotted using GraphPad Prism 9.3.1. After administration of each of the three polypeptide derivative tablets, the plasma concentrations of the three polypeptide derivatives (i.e., PD065, PD187, and PD183) were measured at the above time points. The results are shown in Figure 6.

[0120] Charge-to-mass ratio (CtR) is the ratio of the charge of a charged particle to its mass. The CtR of the ion pair selected for PD065 was 1102.9 / 1074.5, the CtR of the ion pair selected for PD187 was 1153 / 1120.9, and the CtR of the ion pair selected for PD183 was 945.6 / 919.8.

[0121] The results shown in Figure 6 show that although both compounds PD187 and PD183 have excellent weight loss effects, their oral availability is not improved compared to the tablet group containing compound PD065 under the same delivery environment. Unexpectedly, compound PD183 molecule showed a special synergistic effect in the tablets containing the delivery agent PNAC, demonstrating a very high oral bioavailability.

[0122] Although the embodiments of the present application have been described above, the present application is not limited to the above specific embodiments and application fields, and the above specific embodiments are merely examples and teachings, and are not limiting. Under the teachings of this specification, those skilled in the art can also create many forms without departing from the scope protected by the claims of the present application, and all of them are included in the protection scope of the present application.

Claims

1. In an amylin polypeptide analog, a derivative thereof, or a pharmaceutically acceptable salt thereof, comprising the amino acid sequence shown in formula (I), X １ CNTATCATQRLAX １４ FLX １７ HX １９ X ２０ X ２１ X ２２ X ２３ X ２４ X ２５ ILX ２８ X ２９ TNVGSX ３５ TX ３７ Formula (I), Here, X １ is selected from K, D, Dab, Dap, E, H, Har, Orn, R, or deletion. X １４ The value is selected from N, Aad, Dab, Dap, E, or D. X １７ It is selected from V, Orn, or R. X １９ It is selected from S or K, X ２０ It is selected from S or K, X ２１ It is selected from N or K, X ２２ It is selected from N or K, X ２３ It is selected from F, L, or K. X ２４ It is selected from G, R, H, P, D, N, S, K, αK, Dap, Dab, Orn, Cit, Sar, Trx or AOC, X ２５ It is selected from A, K, or P. X ２８ It is selected from S or P, X ２９ It is selected from S or P, X ３５ It is selected from N or G. X ３７ amyrin polypeptide analogs, derivatives thereof, or pharmaceutically acceptable salts thereof, selected from Y, 3-Am-Pro, Aze, cis-Hyp, P, Pip, αMePro, or trans-Hyp.

2. It contains the amino acid sequence shown in formula (II), X １ CNTATCATQRLAX １４ FLX １７ HSSNNX ２３ X ２４ X ２５ ILX ２８ X ２９ TNVGSNTX ３７ Formula (II), Here, X １ It is selected from K or R, X １４ It is selected from N or D, X １７ It is selected from V or R, X ２３ It is selected from F or L, X ２４ It is selected from G, R, or K. X ２５ It is selected from A or P, X ２８ It is selected from S or P, X ２９ It is selected from S or P, X ３７ The polypeptide analog or derivative thereof, or a pharmaceutically acceptable salt thereof, as described in claim 1, selected from Y, P, or trans-Hyp.

3. X １４ is D, X １７ R is X ２３ L is X ２５ P is X ２８ P is X ２９ P is X ３７ A polypeptide analog or derivative thereof according to claim 2, wherein is P or trans-Hyp, or a pharmaceutically acceptable salt thereof.

4. X ２４ is K or R, preferably X １ A polypeptide analog or derivative thereof according to claim 3, wherein is K.

5. A polypeptide analog or derivative thereof according to claim 1, comprising an amino acid sequence selected from SEQ ID NOs: 2 to 104, or a pharmaceutically acceptable salt thereof.

6. The polypeptide analog, derivative thereof, or pharmaceutically acceptable salt thereof according to claim 1, wherein the derivative thereof comprises one to ten amino acid modifications.

7. A polypeptide analog or derivative thereof according to claim 6, or a pharmaceutically acceptable salt thereof, wherein the modification is located at the first K Epsilon amino group.

8. The polypeptide analog, derivative thereof, or pharmaceutically acceptable salt thereof according to claim 6, wherein the amino acid modification involves introducing a functional group that can be used for coupling into the side chain of an amino acid.

9. The functional group that can be used for the coupling comprises an extension portion and a linker that connects the extension portion to the modified amino acid, the polypeptide analog or derivative thereof according to claim 8, or a pharmaceutically acceptable salt thereof.

10. The extended portion is 【Chemistry 1】 Selected from, where x is any integer from 4 to 38, Preferably, The extended portion is HOOC (CH ２ ) １４ CO-, HOOC(CH ２ ) １５ CO-, HOOC(CH ２ ) １６ CO-, HOOC(CH ２ ) １７ CO-, HOOC(CH ２ ) １８ CO-, HOOC(CH ２ ) １９ CO-, HOOC(CH ２ ) ２０ CO-, HOOC(CH ２ ) ２１ CO-, and HOOC (CH ２ ) ２２ A polypeptide analog or derivative thereof, selected from CO-, according to claim 9, or a pharmaceutically acceptable salt thereof.

11. The aforementioned linker is 【Chemistry 2】 【Transformation 3】 【Chemistry 4】 Selected from, where m is 0, 1, 2, or 3, n is 1, 2, or 3, s is any integer from 0 to 6, and p is any integer from 1 to 8. Preferably, the linker is: 【Transformation 5】 A polypeptide analog or derivative thereof according to claim 9, or a pharmaceutically acceptable salt thereof, selected from, where m is 1, n is 1 or 2, and p is any integer from 1 to 5.

12. The polypeptide analog according to claim 11, or a derivative thereof, or a pharmaceutically acceptable salt thereof, is selected from PD069, PD070, PD071, PD072, PD140, PD158, PD160, PD161, PD144, PD145, PD146, PD147, or PD187.

13. The aforementioned derivative or a pharmaceutically acceptable salt thereof is 【Transformation 6】 The polypeptide analog or derivative thereof according to claim 12, or a pharmaceutically acceptable salt thereof.

14. The derivative or a pharmaceutically acceptable salt thereof is 【Transformation 7】 The polypeptide analog or derivative thereof according to claim 12, or a pharmaceutically acceptable salt thereof.

15. The derivative or a pharmaceutically acceptable salt thereof is 【Transformation 8】 The polypeptide analog or derivative thereof according to claim 12, or a pharmaceutically acceptable salt thereof.

16. The derivative or a pharmaceutically acceptable salt thereof is 【Chemistry 9】 The polypeptide analog or derivative thereof according to claim 12, or a pharmaceutically acceptable salt thereof.

17. A method for preparing a polypeptide analog, a derivative thereof, or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 16.

18. Use of a polypeptide analog, derivative thereof, or pharmaceutically acceptable salt thereof according to any one of claims 1 to 16 in the manufacture of a drug for preventing or treating a disease.

19. The use according to claim 18, wherein the disease is an amyrin receptor-related disease.

20. The use according to claim 19, wherein the amyrin receptor-related disease is selected from lipid metabolism disorders, blood glucose metabolism disorders, cardiovascular diseases, and brain, mental or nervous system diseases.

21. A pharmaceutical composition comprising a polypeptide analog or derivative thereof according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof.

22. The pharmaceutical composition according to claim 21, further comprising one or more oral delivery agents, preferably the oral delivery agent being an N-[8-(2-hydroxybenzoyl)amino]octanoic acid (NAC) salt, and more preferably PNAC.

23. The pharmaceutical composition according to claim 21, further comprising one or more other substances having pharmacological activity.

24. The pharmaceutical composition according to claim 23, wherein the substance having pharmacological activity is selected from weight regulators, anti-obesity agents, lipid metabolism regulators, blood glucose regulators, antihypertensive agents, cardiovascular regulators, brain system regulators, psychotropic system regulators, or nervous system regulators.

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