Calcitonin analogue, and derivative thereof and use thereof

Abstract

A calcitonin analogue, and a derivative or a pharmaceutically acceptable salt thereof. A pharmaceutical composition, containing the analogue or the derivative or pharmaceutically acceptable salt thereof, and further comprising a pharmaceutically acceptable excipient. The use of the analogue or the derivative or pharmaceutically acceptable salt thereof and the composition in the field of disease prevention / treatment.

Description

A calcitonin analogue and its derivatives and uses Technical Field

[0001] The present application relates to the field of polypeptide technology, and in particular to a calcitonin analogue and its derivatives and uses. Background Art

[0002] Amylin is a 37-amino acid polypeptide hormone co-secreted with insulin in pancreatic beta cells and is deficient in diabetics. It exerts its effects in several different organ systems, primarily through amylin receptors 1-3 (AMYR1-3). Amylin inhibits glucagon secretion, delays gastric emptying, signals satiety, and suppresses appetite. Clinical studies have shown that amylin receptor agonists are useful for treating overweight, obesity, type 1 diabetes, and / or type 2 diabetes. The amylin analog, pramlintide (trade name Symlin), can lower blood glucose levels and is used to treat diabetic patients taking insulin. However, pramlintide has a half-life of less than an hour and is administered at mealtimes, requiring patients to take multiple doses daily. Pramlintide is unstable and readily fibrillates at neutral pH, rendering it ineffective due to precipitation. Therefore, pramlintide is supplied in an acidic solution.

[0003] Calcitonin is a hormone produced in the thyroid gland that regulates the levels of calcium and phosphate in the blood. Calcitonin receptors are present in many tissues throughout the body and are involved in the regulation of bone metabolism. Salmon calcitonin can be used to treat diseases such as hypercalcemia and osteoporosis, and is currently sold under the trade name Miacalcic. Calcitonin is also unstable at neutral pH, so it is also provided in acidic solutions. The half-life of salmon calcitonin is less than two hours, so patients need to take it once or more daily. The calcitonin drugs currently on the market are salmon calcitonin and elcatonin (eel calcitonin). Due to the relatively small molecular weight of the peptide, it needs to be injected daily. In addition, it is only stable under acidic conditions and has poor stability under neutral conditions, which poses a huge obstacle to the drugability of the molecule and the development of formulations.

[0004] Natural amylin and calcitonin receptor agonists have short half-lives, making extending their half-lives a goal in drug development. Furthermore, amylin and calcitonin receptor agonists are unstable and prone to fibrosis at neutral pH, making improving their stability at physiological pH another goal in drug development.

[0005] Summary of the Invention

[0006] Based on this, the present application improves the amylin receptor agonist polypeptide in order to obtain analogs or derivatives thereof or pharmaceutically acceptable salts with higher in vivo efficacy and greater stability.

[0007] Specifically, this application adopts the following technical solutions:

[0008] A calcitonin derivative or a pharmaceutically acceptable salt thereof, comprising a polypeptide consisting of the following amino acid sequence:

[0009] X0 X1SX3LS TX7VLG RLSX 14 E LHRLX 20 X 21 X 22 PRT X 26 X 27 GX 29 X 30 SX 32 -NH2

[0010] in:

[0011] X0 is D or E or does not exist;

[0012] X1 is A, L, V, I, S, E, R or K;

[0013] X3 is Q, E or D;

[0014] X7 is A, L, V, I, S, R, E or D;

[0015] X 14 is Q, D, E, Aib, S or T;

[0016] X 20 is Q or E;

[0017] X 21 is D or E;

[0018] X 22 Y, L or F;

[0019] X 26 D or Q;

[0020] X 27 V or S;

[0021] X 29 is S or A;

[0022] X 30 is G or E;

[0023] X 32 It is P or trans-Hyp.

[0024] In a preferred embodiment, the calcitonin derivative or a pharmaceutically acceptable salt thereof comprises a polypeptide consisting of the following amino acid sequence:

[0025] X0 X1SX3LS TX7VLG RLSX 14 E LHRLX 20 X 21 X 22 PRT X 26 X 27 GX 29 X 30 SX 32 -NH2

[0026] in:

[0027] X0 is D or E or does not exist;

[0028] The combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, ER, RE, RD, KE, KD or EE;

[0029] X3 is Q, E or D;

[0030] X 14 is Q, D, E, Aib, S or T;

[0031] X 20 is Q or E;

[0032] X 21 is D or E;

[0033] X 22 Y, L or F;

[0034] X 26 D or Q;

[0035] X 27 V or S;

[0036] X 29 -X 30 The combination is selected from SG, AG or AE, SE;

[0037] X 32 It is P or trans-Hyp.

[0038] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is AA, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0039] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is LL, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0040] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is VV, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0041] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is II, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0042] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SS, X3 is Q, E or D; 14is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0043] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is AS, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0044] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SA, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0045] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is ER, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0046] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0047] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RD, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0048] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is KE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0049] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is KD, X3 is Q, E or D; the X 14 is Q, D, E, Aib, S or T; the X20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0050] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is EE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0051] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is AA, X3 is Q; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0052] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is LL, X3 is Q; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32It is P or trans-Hyp.

[0053] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is VV, X3 is Q; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0054] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is II, X3 is Q; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0055] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SS, X3 is Q; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0056] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is AS, X3 is Q; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0057] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SA, X3 is Q; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0058] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is ER, X3 is Q; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0059] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RE, X3 is Q; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0060] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RD, X3 is Q; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0061] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is KE, X3 is Q; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0062] In a preferred embodiment, the X0 does not exist, the combination of the X1 and X7 is KD, the X3 is Q; the X 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0063] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is EE, X3 is Q; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0064] In a preferred embodiment, X0 is absent, the combination of X1 and X7 is AA, or LL, or VV, or II, or SS, or AS, or SA, or ER, or RE, or EE, and X3 is D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0065] In a preferred embodiment, X0 is absent, the combination of X1 and X7 is AA, or LL, or VV, or II, or SS, or AS, or SA, or ER, or RE, or EE, and X3 is E; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0066] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is AA, X3 is Q, E or D; 14 is Q, D or E; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32It is P or trans-Hyp.

[0067] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is LL, X3 is Q, E or D; 14 is Q, D or E; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0068] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is VV, X3 is Q, E or D; 14 is Q, D or E; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q, X 27 is V or S, the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0069] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is II, X3 is Q, E or D; 14 is Q, D or E; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0070] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SS, X3 is Q, E or D; 14 is Q, D or E; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0071] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is AS, X3 is Q, E or D; 14 is Q, D or E; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0072] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SA, X3 is Q, E or D; 14 is Q, D or E; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0073] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is ER, X3 is Q, E or D; 14 is Q, D or E; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0074] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RE, X3 is Q, E or D;14 is Q, D or E; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0075] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RD, X3 is Q, E or D; 14 is Q, D or E; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0076] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is KE, X3 is Q, E or D; 14 is Q, D or E; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0077] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is KD, X3 is Q, E or D; the X 14 is Q, D or E; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X32 It is P or trans-Hyp.

[0078] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is EE, X3 is Q, E or D; 14 is Q, D or E; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0079] In a preferred embodiment, X0 is absent, the combination of X1 and X7 is AA, or LL, or VV, or II, or SS, or AS, or SA, or ER, or RE, or RD, or KE, or KD or EE, X3 is Q, E or D; 14 is Q; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0080] In a preferred embodiment, X0 is absent, the combination of X1 and X7 is AA, or LL, or VV, or II, or SS, or AS, or SA, or ER, or RE, or RD, or KE, or KD or EE, X3 is Q, E or D; 14 is D; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0081] In a preferred embodiment, X0 is absent, the combination of X1 and X7 is AA, or LL, or VV, or II, or SS, or AS, or SA, or ER, or RE, or RD, or KE, or KD or EE, X3 is Q, E or D; 14 is E; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0082] In a preferred embodiment, X0 is absent, the combination of X1 and X7 is AA, or LL, or VV, or II, or SS, or AS, or SA, or ER, or RE, or RD, or KE, or KD or EE, X3 is Q, E or D; 14 is Aib; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0083] In a preferred embodiment, X0 is absent, the combination of X1 and X7 is AA, or LL, or VV, or II, or SS, or AS, or SA, or ER, or RE, or RD, or KE, or KD or EE, X3 is Q, E or D; 14 is S; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0084] In a preferred embodiment, X0 is absent, the combination of X1 and X7 is AA, or LL, or VV, or II, or SS, or AS, or SA, or ER, or RE, or RD, or KE, or KD or EE, X3 is Q, E or D; 14 is T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0085] In a preferred embodiment, X0 is absent, the combination of X1 and X7 is AA, or LL, or VV, or II, or SS, or AS, or SA, or ER, or RE, or RD, or KE, or KD or EE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is E; the X 21 is D or E; the X 22 is Y, L or F; the X 29 -X 30 The combination is selected from SG, AG or AE, SE.

[0086] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is AA, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0087] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is LL, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q; the X 21 is D or E; the X22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0088] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is VV, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0089] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is II, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0090] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SS, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0091] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is AS, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0092] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SA, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0093] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is ER, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0094] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0095] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RD, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0096] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is KE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0097] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is KD, X3 is Q, E or D; the X 14 is Q, D, E, Aib, S or T; the X 20 is Q; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0098] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is EE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0099] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is AA, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0100] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is LL, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0101] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is VV, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is E; the X 22 Y, L or F; X 26 D or Q; X27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0102] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is II, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0103] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SS, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0104] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is AS, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0105] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SA, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0106] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is ER, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0107] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0108] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RD, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is E; the X 22 Y, L or F; X 26 D or Q; X27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0109] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is KE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0110] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is KD, X3 is Q, E or D; the X 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0111] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is EE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0112] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is AA, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0113] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is LL, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0114] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is VV, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0115] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is II, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D; the X 22 Y, L or F; X 26 D or Q; X27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0116] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SS, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0117] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is AS, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0118] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SA, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0119] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is ER, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0120] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0121] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RD, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0122] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is KE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D; the X 22 Y, L or F; X 26 D or Q; X27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0123] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is KD, X3 is Q, E or D; the X 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0124] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is EE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0125] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is AA, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 is Y; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0126] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is LL, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 is Y; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0127] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is VV, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 is Y; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0128] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is II, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 is Y; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0129] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SS, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 is Y; X 26 D or Q; X 27is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0130] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is AS, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 is Y; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0131] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SA, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 is Y; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0132] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is ER, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 is Y; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0133] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RE, X3 is Q, E or D; 14is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 is Y; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0134] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RD, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 is Y; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0135] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is KE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 is Y; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0136] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is KD, X3 is Q, E or D; the X 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 is Y; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X32 It is P or trans-Hyp.

[0137] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is EE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 is Y; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0138] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is AA, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0139] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is LL, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0140] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is VV, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X22 L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0141] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is II, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0142] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SS, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0143] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is AS, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0144] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SA, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0145] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is ER, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0146] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0147] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RD, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 L; X 26 D or Q; X 27is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0148] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is KE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0149] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is KD, X3 is Q, E or D; the X 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0150] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is EE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0151] In a preferred embodiment, X0 is absent; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, ER, RE or EE; X3 is Q or D; X 14 is Q, D, E, Aib, S or T; X 20 Q or E; X 21 D or E; X 22 F; X 26 D or Q; X 27 V or S; X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0152] In a preferred embodiment, X0 is absent; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, ER, RE or EE; X3 is Q or D; X 14 is Q, D, E, Aib, S or T; X 20 Q or E; X 21 D or E; X 22 Y, L or F; X 26 -X 27 Combination: DV; X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0153] In a preferred embodiment, X0 is absent; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, ER, RE or EE; X3 is Q or D; X 14 is Q, D, E, Aib, S or T; X 20 Q or E; X 21 D or E; X 22 Y, L or F; X 26 -X 27 The combination is QS; X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0154] In a preferred embodiment, X0 is absent; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, ER, RE or EE; X3 is Q or D; X 14is Q, D, E, Aib, S or T; X 20 Q or E; X 21 D or E; X 22 Y, L or F; X 26 -X 27 The combination is QV; X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0155] In a preferred embodiment, X0 is absent; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, ER, RE or EE; X3 is Q or D; X 14 is Q, D, E, Aib, S or T; X 20 Q or E; X 21 D or E; X 22 Y, L or F; X 26 -X 27 Combination: DS; X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0156] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is AA, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 The combination is selected from SG; X 32 It is P or trans-Hyp.

[0157] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is LL, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30The combination is selected from SG; X 32 It is P or trans-Hyp.

[0158] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is VV, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 The combination is selected from SG; X 32 It is P or trans-Hyp.

[0159] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is II, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 The combination is selected from SG; X 32 It is P or trans-Hyp.

[0160] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SS, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 The combination is selected from SG; X 32 It is P or trans-Hyp.

[0161] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is AS, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 The combination is selected from SG; X 32 It is P or trans-Hyp.

[0162] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is SA, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 The combination is selected from SG; X 32 It is P or trans-Hyp.

[0163] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is ER, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 The combination is selected from SG; X 32 It is P or trans-Hyp.

[0164] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 The combination is selected from SG; X 32 It is P or trans-Hyp.

[0165] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is RD, X3 is Q, E or D; 14is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 The combination is selected from SG; X 32 It is P or trans-Hyp.

[0166] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is DE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 The combination is selected from SG; X 32 It is P or trans-Hyp.

[0167] In a preferred embodiment, the X0 does not exist, the combination of X1 and X7 is KD, X3 is Q, E or D; the X 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 The combination is selected from SG; X 32 It is P or trans-Hyp.

[0168] In a preferred embodiment, X0 does not exist, the combination of X1 and X7 is EE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 The combination is selected from SG; X 32 It is P or trans-Hyp.

[0169] In a preferred embodiment, X0 is absent; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, ER, RE or EE; X3 is Q or D; X 14 is Q, D, E, Aib, S or T; X 20 Q or E; X 21 D or E; X 22 Y, L or F; X 26 D or Q; X 27 V or S; X 29 -X 30 The combination is selected from AG; X 32 It is P or trans-Hyp.

[0170] In a preferred embodiment, X0 is absent; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, ER, RE or EE; X3 is Q or D; X 14 is Q, D, E, Aib, S or T; X 20 Q or E; X 21 D or E; X 22 Y, L or F; X 26 D or Q; X 27 V or S; X 29 -X 30 The combination is selected from AE; X 32 It is P or trans-Hyp.

[0171] In a preferred embodiment, X0 is absent; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, ER, RE or EE; X3 is Q or D; X 14 is Q, D, E, Aib, S or T; X 20 Q or E; X 21 D or E; X 22 Y, L or F; X 26 D or Q; X 27 V or S; X 29 -X 30 The combination is selected from SE; X 32 It is P or trans-Hyp.

[0172] In a preferred embodiment, X0 is absent; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, ER, RE or EE; X3 is Q or D; X 14is Q, D, E, Aib, S or T; X 20 Q or E; X 21 D or E; X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 For P.

[0173] In a preferred embodiment, X0 is absent; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, ER, RE or EE; X3 is Q or D; X 14 is Q, D, E, Aib, S or T; X 20 Q or E; X 21 D or E; X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 For trans-Hyp.

[0174] In a preferred embodiment, X0 is D; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, ER, RE or EE; X3 is Q or D; X 14 is Q, D, E, Aib, S or T; X 20 Q or E; X 21 D or E; X 22 Y, L or F; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0175] In a preferred embodiment, X0 is E; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, ER, RE or EE; X3 is Q or D; X 14 is Q, D, E, Aib, S or T; X 20 Q or E; X 21 D or E; X 22 Y, L or F; X 26 D or Q; X27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0176] In a preferred embodiment, X0 is D or E or absent; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, RE, RD, KE or KD; X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y or L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0177] In a preferred embodiment, X0 is D or E or absent; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, RE, RD, KE or KD; X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 -X 21 The combination of EE; the X 22 Y or L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0178] In a preferred embodiment, X0 is D or E or absent, the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, RE, RD, KE or KD, X3 is Q, E or D; 14 -X 22 The combination of DL; the X 20 is Q or E; the X 21 D or E; X 26 D or Q; X 27 is V or S; the X 29 -X30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0179] In a preferred embodiment, X0 is D or E or absent, the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, RE, RD, KE or KD, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y or L; X 26 -X 27 is QS; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0180] In a preferred embodiment, X0 is D or E or absent, the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, RE, RD, KE or KD, X3-X 14 -X 20 is EEE; the X 21 is D or E; the X 22 Y or L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0181] In a preferred embodiment, X0 is D or E or absent; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, RE; X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y or L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0182] In a preferred embodiment, X0 is D or E or absent; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, RE; X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 -X 21 is EE; the X 22 Y or L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0183] In a preferred embodiment, X0 is D or E or absent; the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, RE; X3 is Q, E or D; 14 -X 22 is DL; the X 20 is Q or E; the X 21 D or E; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0184] In a preferred embodiment, X0 is D or E or absent, the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, RE, X3 is Q, E or D; 14 is Q, D, E, Aib, S or T; the X 20 is Q or E; the X 21 is D or E; the X 22 Y or L; X 26 -X 27 is QS; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0185] In a preferred embodiment, X0 is D or E or absent, and the combination of X1 and X7 is selected from AA, LL, VV, II, SS, AS, SA, RE, X3-X 14 -X 20 is EEE; the X 21 is D or E; the X 22 Y or L; X 26 D or Q; X 27 is V or S; the X 29 -X 30 A combination of SG, AG or AE, SE; X 32 It is P or trans-Hyp.

[0186] In a preferred embodiment, the derivative or its pharmaceutically acceptable salt contains a polypeptide consisting of the following amino acid sequence: ASQLS TAVLG RLSX 14 E LHRLX 20 DX 22 PRT DVGX 29 G SP-NH2, in which

[0187] X 14 is Q, D or E;

[0188] X 20 Q or E, preferably Q;

[0189] X 22 is Y or L;

[0190] X 29 It is S or A.

[0191] In a preferred embodiment, the derivative or its pharmaceutically acceptable salt contains a polypeptide consisting of the following amino acid sequence: ASQLS TAVLG RLSX 14 E LHRLQ DX 22 PRT DVGSG SP-NH2, where

[0192] X 14 Q, D or E, preferably Q or D;

[0193] X 22 It is Y or L.

[0194] In a preferred embodiment, the amino acid sequence of the calcitonin derivative or a pharmaceutically acceptable salt thereof is selected from the sequences in Table 2 below: any one of SEQ ID NO.1 to SEQ ID NO.38.

[0195] In a preferred embodiment, the amino acid sequence of the calcitonin derivative or pharmaceutically acceptable salt is selected from any one of the following sequences:

[0196] ASQLS TAVLG RLSDE LHRLQ DYPRT DVGSG SP-NH2(SEQ ID NO.7),

[0197] ASQLS TAVLG RLSQE LHRLQ DLPRT DVGSG SP-NH2(SEQ ID NO.17),

[0198] ASQLS TAVLG RLSEE LHRLQ DYPRT DVGSG SP-NH2(SEQ ID NO.8),

[0199] ASQLS TAVLG RLSQE LHRLQ DYPRT DVGAG SP-NH2(SEQ ID NO.18),

[0200] ASQLS TAVLG RLSQE LHRLQ DYPRT DVGAE SP-NH2 (SEQ ID NO.19) or

[0201] ASQLS TAVLG RLSQE LHRLE DYPRT DVGSG SP-NH2 (SEQ ID NO. 12).

[0202] In a preferred embodiment, the calcitonin derivative or a pharmaceutically acceptable salt thereof comprises a side chain of a modified amino acid; preferably, the side chain is a fatty acid-containing side chain.

[0203] In a preferred embodiment, the calcitonin derivative or a pharmaceutically acceptable salt thereof comprises a fatty acid-containing side chain connected to the N-terminus of its amino acid sequence.

[0204] In a preferred embodiment, the fatty acid-containing side chain is in the form of Z1+Z2+Z3, wherein Z1 is a C16-C22 fatty diacid;

[0205] Z2 is selected from any one of γGlu, αGlu, βAsp, αAsp, Inp, Trx or Z2 does not exist;

[0206] Z3 is n AEEAs, where n≥0.

[0207] The Z1, Z2 and Z3 are sequentially connected via amide bonds.

[0208] In a preferred embodiment, n is selected from 0, 1, 2, 3, 4, 5 or 6.

[0209] In a preferred embodiment, n is 6.

[0210] In a preferred embodiment, n is 4.

[0211] In a preferred embodiment, n is 2.

[0212] In a preferred embodiment, n is 0.

[0213] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C16-C22 fatty diacid) + γGlu.

[0214] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C16-C22 fatty diacid) + αGlu.

[0215] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C16-C22 fatty diacid) + βAsp.

[0216] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C16-C22 fatty diacid) + αAsp.

[0217] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C16-C22 fatty diacid)+Inp.

[0218] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C16-C22 fatty diacid)+Trx.

[0219] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C16-C22 fatty diacid).

[0220] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C16-C22 fatty diacid)+γGlu+2AEEA.

[0221] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C16-C22 fatty diacid)+γGlu+4AEEA.

[0222] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C16-C22 fatty diacid)+γGlu+6AEEA.

[0223] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C16-C22 fatty diacid)+αGlu+2AEEA.

[0224] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C16-C22 fatty diacid)+βAsp+2AEEA.

[0225] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C16-C22 fatty diacid)+αAsp+2AEEA.

[0226] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C16-C22 fatty diacid)+Inp+2AEEA.

[0227] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C16-C22 fatty diacid)+Trx+2AEEA.

[0228] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C20 fatty diacid)+γGlu.

[0229] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C20 fatty diacid)+αGlu.

[0230] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C20 fatty diacid)+βAsp.

[0231] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C20 fatty diacid)+αAsp.

[0232] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C20 fatty diacid)+Inp.

[0233] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C20 fatty diacid)+Trx.

[0234] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C20 fatty diacid).

[0235] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C20 fatty diacid)+γGlu+2AEEA.

[0236] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C20 fatty diacid)+γGlu+4AEEA.

[0237] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C20 fatty diacid)+γGlu+6AEEA.

[0238] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C20 fatty diacid)+αGlu+2AEEA.

[0239] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C20 fatty diacid)+βAsp+2AEEA.

[0240] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C20 fatty diacid)+αAsp+2AEEA.

[0241] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C20 fatty diacid)+Inp+2AEEA.

[0242] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C20 fatty diacid)+Trx+2AEEA.

[0243] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C22 fatty diacid)+γGlu+2AEEA.

[0244] In a preferred embodiment, the fatty acid-containing side chain is in the form of (C22 fatty diacid)+γGlu+4AEEA.

[0245] The present application also provides a polypeptide analog, which is a polypeptide containing the amino acid sequence of any one of the above derivatives.

[0246] The present application also provides a pharmaceutical composition comprising one or more of the analogs, derivatives, or pharmaceutically acceptable salts thereof described above. Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient, such as a non-toxic filler, stabilizer, diluent, carrier, solvent, or other formulation excipient.

[0247] The present application also provides the use of the above-mentioned analogs, derivatives or pharmaceutically acceptable salts thereof in the preparation of a medicament for preventing and / or treating overweight, obesity and / or type I or type II diabetes and / or osteoporosis and / or neuropathic pain.

[0248] The present application also provides the use of the analogs, derivatives, or pharmaceutically acceptable salts thereof described above for reducing food intake and / or weight loss. For example, the use of the analogs, derivatives, or pharmaceutically acceptable salts thereof described above for preparing health products, foods, or medicines for reducing food intake and / or weight loss.

[0249] The present application also provides a method for preventing and / or treating overweight, obesity and / or type I or type II diabetes and / or osteoporosis and / or neuropathic pain, comprising administering to a subject an effective amount of the analogs, derivatives or pharmaceutically acceptable salts thereof, or pharmaceutical compositions described above in the present application.

[0250] The present application also provides a method for reducing food intake, comprising administering to a subject an effective amount of the analogs, derivatives or pharmaceutically acceptable salts thereof, or pharmaceutical compositions described above in the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0251] The accompanying drawings are provided to facilitate a better understanding of the present application and do not constitute an undue limitation on the present application.

[0252] Figure 1 is a reverse phase chromatogram of D3 (Mimylin) molecule after thermal acceleration for 26 days;

[0253] FIG2 is a reverse phase chromatogram of a calcitonin analog (M5) after heat acceleration for 26 days;

[0254] FIG3 is a reverse phase chromatogram of M8 single molecule thermally accelerated under neutral conditions for 26 days;

[0255] FIG4 is a reverse phase chromatogram of M8 and M51 after heat acceleration for one month in a neutral buffer without excipients;

[0256] FIG5 is a reverse phase chromatogram of M8 and M51 after heat acceleration for one month in a neutral buffer with excipients;

[0257] FIG6 is a trend diagram showing the effects of various calcitonin analogs or derivatives on changes in rat body weight in the first group of experiments;

[0258] FIG7 is a trend diagram showing the effects of various calcitonin analogs or derivatives on the cumulative food intake of rats in the first group of experiments;

[0259] FIG8 is a trend diagram showing the effects of various calcitonin analogs or derivatives on changes in rat body weight in the second group of experiments;

[0260] FIG9 is a trend diagram showing the effects of various calcitonin analogs or derivatives on the cumulative food intake of rats in the second group of experiments. DETAILED DESCRIPTION

[0261] The "analog" mentioned herein refers to a compound in which at least one amino acid residue of a specific polypeptide molecule has been replaced by another amino acid residue, and / or at least one amino acid residue has been deleted, and / or at least one amino acid residue has been added to the N-terminus or C-terminus, and / or at least one amino acid residue has been inserted between any two amino acids.

[0262] Unless the context clearly indicates otherwise, the terms "polypeptide," "analog," and combinations thereof, used herein, have the same meaning and are interchangeable. In certain circumstances, the terms "calcitonin" and "calcitonin analog" have the same meaning and are interchangeable, for example, in "derivative of a calcitonin analog" and "derivative of calcitonin."

[0263] As used herein, "derivatives" refer to products obtained by modifying functional groups (e.g., amino acid residues) of biological macromolecules (e.g., polypeptides and proteins) with certain groups or structures (e.g., certain small molecule structures). Such modifications include, but are not limited to, acylation, amidation, esterification, and thioesterification.

[0264] abbreviation

[0265] The corresponding names or structural forms of some abbreviations used in the examples of this application are as follows:

[0266] 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-Pentamet hyldihydrobenzofuran-5-sulfonyl chloride; Pip:

[0267] Piperidine, piperidine; TBTU: O-(Benzotriazol-l-yl)-N,N,N,N,-tetramethyluroniumTetrafluoroborate, O-Benzotriazol-N,N,N',N'-tetramethyluroniumtetrafluoroborate; tBu: tertiary butyl; TFA: Trifluoroacetic acid; TIS: Triisopropylsilane; Trt: Triphenylmethyl; αGlu: α-glutamic acid; γGlu: γ-glutamic acid; αAsp: α-aspartic acid; βAsp: β-aspartic acid;

[0268] The following description of exemplary embodiments of the present application includes various details of the embodiments of the present application to facilitate understanding, and should be considered as merely exemplary. Therefore, those skilled in the art will recognize that various changes and modifications may be made to the embodiments described herein without departing from the scope and spirit of the present application. Similarly, for the sake of clarity and conciseness, descriptions of well-known functions and structures are omitted in the following description.

[0269] Example

[0270] Example 1 Preparation of polypeptide derivatives

[0271] Using solid-phase organic synthesis, Fmoc-protected amino acids, and SPPS solid-phase synthesis technology, peptide synthesis, cleavage, oxidation, and purification were completed to obtain the target product. Using compound Cagrilintide (No. D1) as an example, the synthesis process is as follows.

[0272] 1.1 Solid-phase synthesis

[0273] Using Fmoc-Linker MBHA Resin S = 0.32 mmol / g and Fmoc / tBu technology, amino acids were sequentially condensed and linked from the C-terminus to the N-terminus (from right to left) according to the above peptide sequence using the method in Table 1.

[0274] Table 1 Synthesis procedures

[0275] The following amino acids were coupled sequentially:

[0276] 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-Pr o-OH,A-11Fmoc-Leu-OH,A-12Fmoc-Ile-OH,A-13Fmoc-Pro-OH,A-14Fmoc-Gly-OH,A-15Fmoc-Phe-OH,A-16F moc-Asn(Trt)-OH,A-17Fmoc-Asn(Trt)-OH,A-18Fmoc-Ser(tBu)-OH,A-19Fmoc-Ser(tBu)-OH,A-20Fmoc-His (Trt)-OH,A-21Fmoc-Arg(Pbf)-OH,A-22Fmoc-Leu-OH,A-23Fmoc-Phe-OH,A-24Fmoc-Glu(OtBu)-OH,A-25Fm oc-Ala-OH,A-26Fmoc-Leu-OH,A-27Fmoc-Arg(Pbf)-OH,A-28Fmoc-Gln(Trt)-OH,A-29Fmoc-Thr(tBu)-OH,A- 30Fmoc-Ala-OH,A-31Fmoc-Cys(Trt)-OH,A-32Fmoc-Thr(tBu)-OH,A-33Fmoc-Ala-OH,A-34Fmoc-Thr(tBu)- OH,A-35Fmoc-Asn(Trt)-OH,A-36Fmoc-Cys(Trt)-OH,A-37Fmoc-Lys(Boc)-OH,A-38Fmoc-Glu-otbu,A-39C20 diacid;

[0277] Finally, the polypeptide derivative resin is formed.

[0278] The peptide derivative resin is washed, transferred out, dried to constant weight, and then cracked.

[0279] 1.2 Cracking

[0280] Preparation of cleavage reagent: Calculate the cleavage reagent dosage based on a ratio of TFA:H₂O:EDT:TIS (95:1:2:2) per 1g of peptide resin. Add the required cleavage reagents, H₂O, TFA, EDT, and TIS, to the cleavage reaction flask in this order. Keep the cleavage reagent temperature between 0 and 10°C. Add the cleavage reagents to the peptide resin with stirring. Once the system temperature stabilizes, continue stirring at 25-30°C for 2.5 hours. Filter the lysate and precipitate with 5 times the liquid volume of glacial ether. Filter the precipitate and wash it three times with 3 times the liquid volume of glacial ether. Dry under reduced pressure at room temperature to obtain a crude solid.

[0281] 1.3 Oxidation

[0282] Grind the crude product into powder, prepare purified water, slowly add the ground crude product while stirring, and at the same time add acetonitrile aqueous solution dropwise. After the crude product is added and completely dissolved, add iodine methanol solution and stir for half an hour.

[0283] 1.4 Purification and freeze-drying

[0284] The oxidized liquid was filtered through a 0.45 μm microporous membrane. The crude product was purified using a C-18 preparative column using an appropriate gradient at room temperature. The target product was collected, analyzed, tested, and classified. Purity was required to be ≥90%. Unqualified target products were collected and further separated and purified using an appropriate gradient. The qualified liquid peak was collected. The qualified liquid sample was freeze-dried under reduced pressure to obtain a powdered, purified polypeptide lyophilized powder.

[0285] The polypeptides and derivatives prepared in this application are shown in Table 2 and can be prepared using methods similar to those of this Example. D2 represents the original salmon calcitonin sequence, D1 and D3 represent modified calcitonin sequences from the prior art, and the others represent calcitonin analogs or derivatives provided in this application. The fatty diacid portion of the polypeptide derivative is located outside the fatty acid-containing side chain used for modification relative to the amino acid sequence, distal to the end where the side chain is connected to the amino acid sequence.

[0286] Table 2 Amino acid sequence and modification

[0287] Example 2 Cellular activity detection of derivatives

[0288] The purpose of this study was to test the activity or efficacy of calcitonin analogs or derivatives at human amylin receptor in vitro using luciferase assay.

[0289] 2.1 Construction of the Amylin Receptor / CRE-luc Cell Line

[0290] Using standard methods, CHO-K1 / Ga15 / AMY3 cells (calcitonin receptor and receptor-activating modified peptide RAMP have been constructed and purchased from GenScript) were transfected with a plasmid containing multiple copies of a cAMP response element (CRE)-driven luciferase expression cassette and cultured in F12 medium containing Zeocin 200 ug / mL, puromycin (2 ug / mL), hygromycin (100 ug / mL) and G418 (400 ug / mL) to obtain a stably transfected amylin receptor / CRE-luc cell line.

[0291] 2.2 Amylin-luciferase assay

[0292] The lyophilized powder obtained according to the method of Example 1 was dissolved in 20 mM phosphate buffer at pH 7.0 and diluted with growth medium (F12 medium containing 10% FBS) to obtain a derivative sample with an initial concentration of 10 nM. The derivative sample was then gradiently diluted with growth medium to obtain seven concentrations of samples with a concentration difference of 5 times from 10 nM to 2 nM-0.4 nM. 50 μL of the sample assay solution of the corresponding concentration was added to each well of a white 96-well plate.

[0293] CHO cells stably transfected with amylin / CRE-luc were resuspended in growth medium at a defined density. 50 μL of the cell suspension was added to a white 96-well plate containing the sample assay solution at a density of approximately 20,000 cells / well. After incubation at 37°C and 5% CO₂ for 24 hours, 100 μL of Luciferase substrate was added to each well and incubated for 3 minutes. Luminescence was measured on a SpectraMax L (Molecular Devices) using SoftMax Pro 7.0.3 GxP software. A standard curve was constructed using the fluorescence values, and the EC50 was calculated. The results are shown in Table 3 below.

[0294] Table 3 Amylin receptor cell activity results

[0295] It can be seen from the data in the table that most of the calcitonin analogs or derivatives provided in the present application have relatively high amylin receptor cell activity, and some derivatives are even one order of magnitude higher than the calcitonin analogs or derivatives in the prior art.

[0296] Example 3 Thermal Accelerated Stability Test

[0297] 3.1 Materials and Equipment

[0298] Stability test chamber (BINDER GmbH), 1 / 100,000 balance (Mettler-Toledo), pH meter (Mettler-Toledo), biological safety cabinet (ESCO), T2G-Ⅱ small electric capping machine (Changsha Zhongya Pharmaceutical Equipment Co., Ltd.), high-speed refrigerated centrifuge (Eppendorf), sterilization cabinet (Xinhua Medical), Agilent 1260 high-performance liquid chromatograph, Sepax Bio-C18 4.6*250mm 3μm Reversed-phase chromatography columns.

[0299] Reagents and consumables: ultrapure water (18.2 MΩ self-made ultrapure water), acetonitrile (HPLC grade), trifluoroacetic acid (HPLC grade), sodium dihydrogen phosphate (pharmaceutical grade / Hunan Jiudian Hongyang), hydrochloric acid (pharmaceutical grade / Hunan Erkang), sodium hydroxide (analytical grade / National Pharmaceutical Shanghai Testing), vials, rubber stoppers, disposable syringes, sterile filters.

[0300] 3.2 Experimental methods

[0301] The lyophilized powder of the calcitonin analog or derivative prepared according to the method of Example 1 was mixed with sodium dihydrogen phosphate (1.42 mg / ml) at a mass-to-volume ratio of 1 mg / ml and dissolved in ultrapure water. The pH was adjusted to approximately 7.4 using hydrochloric acid / sodium hydroxide. The solution was then filtered through a 0.22 μm sterile filter in a clean bench and transferred to a sterile vial. The vial was capped and placed in a 40°C stability chamber. Testing was performed on days 7 and 26. During the experiment, changes in the properties of the peptide were observed and recorded. The sample was then centrifuged at 10,000 rpm, 4°C, for 3 minutes. The supernatant was transferred to a liquid chromatography vial and assayed for peptide concentration and purity using the following liquid chromatography method. Peptide concentration = peak area / sample load / extinction coefficient / 60 × flow rate. Peptide purity = target peak area / total peak area × 100%.

[0302] Reversed-phase chromatography conditions: flow rate: 1.0 ml / min; autosampler temperature: 15°C; column temperature: 25°C; detection wavelengths: 280 nm and 214 nm; mobile phase A: 100% H2O + 0.05% TFA; mobile phase B: 100% ACN; elution gradient: see Table 4 below:

[0303] Table 4 Liquid chromatography elution gradient list

[0304] 3.3 Experimental Results

[0305] The results are shown in Tables 5, 6, and 7 below. It is generally considered (as per the 2020 Pharmacopoeia) that a difference of 5% or more from the initial value is considered a significant change. Within a reasonable error range, if the decrease in the measured content is less than 1% from the initial value, it is indicated as "no change."

[0306] Table 5 Changes in properties of thermally accelerated stability

[0307] As can be seen from Table 5, the calcitonin analogs or derivatives provided by the present application do not show significant changes in thermally accelerated stability under neutral conditions. On the 7th day of the accelerated experiment, the solution remained clear, and the properties of the polypeptide remained essentially unchanged. Furthermore, during the experiment, it was found that the properties of molecules such as M8, M9, and M13 remained unchanged on the 26th day of the accelerated experiment, and the solution remained clear, indicating very good stability.

[0308] Table 6 Concentration changes of thermally accelerated stability

[0309] As can be seen from Table 6, the concentration changes of the calcitonin analogs or derivatives provided by the present application under neutral conditions are not obvious. On the 7th day of the accelerated experiment, the concentration changes were all below 4%. Moreover, during the experiment, it was found that the concentration changes of some calcitonin analogs or derivatives, such as M5, were close to 1%, while some, such as M8 and M13, reached below 1% (marked as no change). Moreover, on the 26th day of the accelerated experiment, the concentration change was also below 2% compared to the initial day 0, indicating very good concentration stability.

[0310] Table 7 Purity Changes in Thermally Accelerated Stability (Reversed Phase Purity - 214nm)

[0311] As can be seen from Table 7, the purity changes of the calcitonin analogs or derivatives provided by the present application under neutral conditions of thermal acceleration stability are not obvious. On the 7th day of the accelerated experiment, the reversed-phase purity changes of most polypeptide solutions were less than 1%, and even no concentration reduction was observed. Moreover, during the experiment, it was found that the purity changes of molecules such as M5, M8, M9, and M13 were even lower on the 26th day of the accelerated experiment, and there was almost no significant change, indicating very good purity stability. Compared with the molecule M3 in the prior art, which decreased in purity by 18% after 26 days of thermal acceleration (see Figure 1), the purity of the calcitonin analog (M5) of the present application decreased by only 0.68% after 26 days of thermal acceleration (see Figure 2). The purity changes of the calcitonin derivative M8 provided by the present application in the thermal acceleration experiment are compared with D1 (Novo Nordisk, Cagrlintide) and are listed in Table 8. The reversed-phase spectra of the molecule M8 before and after thermal acceleration are shown in Figure 3. The data in Table 8 and Figure 3 can also more clearly show the difference between the M8 skeleton and D1, and its stability under neutral conditions is very good.

[0312] Table 8 Comparison of liquid phase analysis results after 26 days of thermal acceleration

[0313] It can be seen from the data of this application that the molecules of the calcitonin analogs and derivatives provided by this application have very high thermal accelerated stability compared with the prior art.

[0314] Example 4 Compatibility Thermal Acceleration Stability Test

[0315] Calcitonin analog M8 (1 mg / ml), GLP-1 receptor agonist M51 (1 mg / ml) and sodium dihydrogen phosphate (1.42 mg / ml) were mixed and dissolved in ultrapure water. 5.5 mg / ml phenol and 14 mg / ml 1,2-propylene glycol were added to the excipient group. The pH was adjusted to about 7.4 with hydrochloric acid / sodium hydroxide. An accelerated stability test was then performed according to the method of Example 3. The experimental results are shown in Figure 4 (without excipients) and Figure 5 (with excipients). It can be seen from the figure that M8 is very compatible with M51. Under the same formulation conditions as M51, the reverse phase spectrum changes very little after one month of thermal acceleration. It has the potential to be developed as a co-formulation. Taking the molecule M8 of the present application as an example, from the current results, the calcitonin analog or derivative of the present application is very stable, both in terms of thermal stability as a single drug and in the same formulation conditions as M51.

[0316] Example 5 Animal Weight Change and Food Consumption Experiment

[0317] SD (Sprague Dawley) rats that select body weight to be 200-250g are used for this experimental study. Before the experiment started, rats arrived at least 10-14 days to adapt to the experimental environment. During this period, rats were raised for two weeks under the light / dark phase (meaning that light is closed during daytime and opened at night) of reversal after arrival, and were raised separately in the first week of arrival to ensure that data are accurate and high in test sensitivity. In the whole adaptation period and experimental period, rats were free to obtain food and water. Each derivative test group of 5-8 rats was given the derivative or vehicle of 10nmol / kg dosage of each group of rats subcutaneously. The administration time of each group was recorded. After administration, rats were returned to their living cage, and then they can obtain food and water therein. By online recording or manual recording, food consumption and rat weight changes were recorded every 24 hours.

[0318] The results of the first set of experiments are shown in Figures 6 and 7, and the results of the second set of experiments are shown in Figures 8 and 9. As can be seen from the figures, the calcitonin analogs and derivatives of the present application have very significant effects in reducing the body weight and food intake of rats.

[0319] As can be further seen from Figures 6 and 7, the calcitonin analogs of the present application all have high weight loss effects and appetite suppression effects, and the effects are better than D1, especially M18, M9, M13, etc. The effects are particularly obvious.

[0320] As can be further seen from FIG8 and FIG9 , the calcitonin analogs and derivatives of the present application, such as M8, M19, and M20, are particularly effective in reducing the body weight or food intake of rats.

[0321] Although the embodiments of the present application are described above, the present application is not limited to the above-mentioned specific embodiments and application fields. The above-mentioned specific embodiments are merely illustrative and instructive, and not restrictive. A person of ordinary skill in the art, under the guidance of this specification and without departing from the scope of protection of the claims of this application, can also make many forms, all of which fall within the scope of protection of this application.

Claims

1. A calcitonin derivative or a pharmaceutically acceptable salt thereof, comprising a polypeptide consisting of the following amino acid sequence: X 0 X 1 SX 3 LS TX 7 VLG RLSX 14 E LHRLX 20 X 21 X 22 PRT X 26 X 27 GX 29 X 30 S X 32 -NH 2 in ： X 0 D or E or not present; X 1 is A, L, V, I, S, E, R or K; X 3 is Q, E or D; X 7 is A, L, V, I, S, R, E or D; X 14 is Q, D, E, Aib, S or T; X 20 is Q or E; X 21 is D or E; X 22 is Y, L or F; X 26 D or Q; X 27 V or S; X 29 is S or A; X 30 is G or E; X 32 It is P or trans-Hyp.

2. The derivative according to claim 1 or a pharmaceutically acceptable salt thereof, in, X 1 and X 7 A combination of AA, LL, VV, II, SS, AS, SA, ER, RE, RD, KE, KD or EE; X 29 -X 30 The combination is selected from SE, SG, AG or AE.

3. The derivative according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, in, X 3 For Q.

4. The derivative according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, in, X 14 It is Q, D or E.

5. The derivative according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, in, X 20 For Q.

6. The derivative according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, in, X 21 is D.

7. The derivative according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, in, X 22 It is Y or L.

8. The derivative according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof, in, X 26 and X 27 The combination is selected from DV, QS, QV, DS.

9. The derivative according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof, in, X 29 -X 30 The combination is SG.

10. The derivative according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof, in, X 32 For P.

11. The derivative according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, in, The derivative or its pharmaceutically acceptable salt contains a polypeptide consisting of the following amino acid sequence: ASQLS TAVLG RLSX 14 ELHRLX 20 DX 22 PRT DVGX 29 G SP-NH 2 ,in, X 14 is Q, D or E; X 20 Q or E, preferably Q; X 22 is Y or L; X 29 It is S or A.

12. The derivative according to claim 11 or a pharmaceutically acceptable salt thereof, in, The derivative or its pharmaceutically acceptable salt contains a polypeptide consisting of the following amino acid sequence: ASQLS TAVLG RLSX 14 E LHRLQ DX 22 PRT DVGSG SP-NH 2 ,in, X 14 Q, D or E, preferably Q or D; X 22 It is Y or L.

13. The derivative or pharmaceutically acceptable salt thereof according to claim 1, wherein the amino acid sequence is selected from any one of SEQ ID NO. 1 to SEQ ID NO.

38.

14. The derivative according to any one of claims 1 to 8 or 10 to 13 or a pharmaceutically acceptable salt thereof, in, The polypeptide sequence is any one of the following sequences: ASQLS TAVLG RLSDE LHRLQ DYPRT DVGSG SP-NH 2 、 ASQLS TAVLG RLSQE LHRLQ DLPRT DVGSG SP-NH 2 、 ASQLS TAVLG RLSEE LHRLQ DYPRT DVGSG SP-NH 2 、 ASQLS TAVLG RLSQE LHRLQ DYPRT DVGAG SP-NH 2 、 ASQLS TAVLG RLSQE LHRLQ DYPRT DVGAE SP-NH 2 or ASQLS TAVLG RLSQE LHRLE DYPRT DVGSG SP-NH 2 。 15. The derivative according to any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof, in, The derivative comprises a fatty acid-containing side chain attached to the N-terminus of the polypeptide.

16. The derivative or pharmaceutically acceptable salt thereof according to claim 15, wherein the side chain containing fatty acid is in the form of Z1+Z2+Z3, in, Z1 is a C16-C22 fatty diacid; Z2 is selected from any one of γGlu, αGlu, βAsp, αAsp, Inp, Trx or Z2 is absent; Z3 is n AEEAs, where n ≥ 0; The Z1, Z2 and Z3 are sequentially connected via amide bonds.

17. The derivative or pharmaceutically acceptable salt thereof according to claim 16, wherein Z1 is a C20 fatty diacid.

18. The derivative or pharmaceutically acceptable salt thereof according to claim 16 or 17, wherein Z2 is γGlu. 19 . The derivative or pharmaceutically acceptable salt thereof according to claim 16 , wherein n is selected from 0, 1, 2, 3, 4, 5 or 6.

20. A polypeptide comprising the amino acid sequence of the derivative according to any one of claims 1 to 14.

21. A pharmaceutical composition comprising the derivative according to any one of claims 1 to 19 or the polypeptide according to claim 20 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

22. Use of the derivative according to any one of claims 1 to 19 or the polypeptide according to claim 20 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating overweight, obesity and / or type I or type II diabetes and / or osteoporosis and / or neuropathic pain.

23. Use of the derivative according to any one of claims 1 to 19 or the polypeptide according to claim 20 or a pharmaceutically acceptable salt thereof in reducing food intake.

24. A method for preventing and / or treating overweight, obesity and / or type I or type II diabetes and / or osteoporosis and / or neuropathic pain, comprising administering to a subject an effective amount of the derivative according to any one of claims 1 to 19, or the polypeptide according to claim 20 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 21.

25. A method for reducing food intake, comprising administering to a subject an effective amount of the derivative according to any one of claims 1 to 19, or the polypeptide according to claim 20 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 21.