Novel polypeptide formulations and therapeutic uses thereof

Abstract

This invention relates to the field of polypeptide chemistry, and more specifically to a polypeptide compound pharmaceutical preparation and its use in medicine.

Description

Technical Field

[0001] This invention relates to a polypeptide compound formulation and its use in medicine. Background of the Invention

[0003] Metabolic syndrome is a combination of medical conditions that increase the risk of developing type 2 diabetes, atherosclerotic vascular disease, heart disease, and stroke. Medical parameters defining metabolic syndrome include diabetes, impaired glucose tolerance, elevated fasting blood glucose, insulin resistance, central obesity, hypertension, elevated total cholesterol and triglycerides, elevated LDL cholesterol, and decreased HDL cholesterol. Diabetes includes type 1 diabetes, type 2 diabetes, and gestational diabetes. According to the World Health Organization (WHO), the prevalence of diabetes in developed countries is 5%–10%, and by 2030, the number of people with diabetes worldwide will double compared to 2000. More than 50% of people worldwide are undiagnosed with diabetes, and the number of people with prediabetes is even greater than the number of people with diabetes. For example, China has 114 million people with diabetes, and another 500 million people with impaired glucose tolerance and impaired glucose regulation are about to become diabetic. More than half of patients are unaware that they have the disease. The greatest harm of diabetes lies mainly in its serious complications and high mortality rate. Data shows that diabetes is the leading cause of lower limb amputation and new-onset blindness in adults.

[0004] Obesity is a medical condition; the accumulation of excess body fat can adversely affect health and life expectancy, and due to its increasing prevalence in adults and children, it has become one of the leading causes of preventable death in modern times. It increases the likelihood of various other diseases, including heart disease, type 2 diabetes, obstructive sleep apnea, certain types of cancer, and osteoarthritis. It is typically caused by a combination of factors, including excessive food intake, reduced energy expenditure, and genetic predisposition. At the same body mass index (BMI), Asians have higher visceral fat levels than Caucasians, resulting in greater insulin resistance. Asians with normal weight and type 2 diabetes have significantly lower insulin sensitivity than non-diabetic individuals. Excess fat leads to insulin resistance and damage to beta cells, impairing glucose regulation. Multiple obesity-related metabolic abnormalities significantly increase the risk of cardiovascular disease. Clinical statistics show that over 70% of type 2 diabetes cases are caused by being overweight. Therefore, reducing weight and body fat in diabetic patients is a crucial way to effectively control and even reverse the progression of diabetes. However, existing small-molecule weight-loss drugs have significant side effects. Glucagon-like peptide-1 (GLP-1) receptor agonists control blood sugar by promoting insulin secretion, improving insulin sensitivity, and reducing glucagon release. Therefore, GLP-1 drugs are suitable for treating metabolic diseases, especially diabetes. GLP-1 receptor agonists such as exenatide and liraglutide have shown weight-reducing effects in animal studies and clinical trials, with relatively few side effects. Liraglutide has been approved in the United States for the treatment of diabetes and obesity, making it the only drug approved for these two conditions.

[0005] However, existing GLP-1 inhibitors all have significant gastrointestinal side effects. These side effects affect patient adherence, reducing the dosage and user base. Based on the physiological mechanism of the GLP-1 receptor, GLP-1 drugs have a slow effect on weight loss, requiring higher doses than those used for diabetes treatment in clinical practice, leading to more pronounced gastrointestinal side effects. Most patients experience an average weight loss of less than 5%, and weight rebound is significant after discontinuation of the medication.

[0006] Therefore, there is a clinical need for drugs that can lower blood sugar, reduce blood lipids and body fat, and reduce weight.

[0007] Patients with diabetes have a higher risk of cardiovascular disease; therefore, they need to strictly control their blood lipid levels. Clinical studies have shown that long-term use of statins may increase the risk of diabetes. Statins and fibrates have significant side effects and both have individuals who are intolerant. Currently, there are no ideal treatments for non-alcoholic fatty liver disease. The polypeptide compounds of this invention can not only lower blood sugar but also significantly reduce triglycerides and total cholesterol, especially low-density cholesterol (LDL), potentially addressing both high blood sugar and high blood lipids in the "three highs" simultaneously, and are expected to have significant benefits for the cardiovascular health of users. The polypeptides of this invention are more suitable for diabetic patients or those with prediabetes than statins. The polypeptides of this invention can provide new treatment options for these diseases. The polypeptides of this invention are applicable to various diseases caused by abnormal lipid metabolism, including hyperlipidemia and non-alcoholic fatty liver disease. The polypeptides of this invention can also be used for hypertension, arteriosclerosis, coronary heart disease, peripheral artery disease, stroke, or any combination of these diseases.

[0008] Most diabetic patients are middle-aged or elderly and need to take multiple medications. This inevitably involves the compatibility and matching of drugs. Besides drug cross-reactions and the potential for increased toxic side effects from multiple medications, the different durations of action and frequencies of administration of different drugs also add to the inconvenience for patients. Therefore, the polypeptides of this invention not only enhance efficacy and reduce toxic side effects but also provide convenience for patients, thereby improving treatment outcomes.

[0009] The peptides of this invention can also be used to treat diabetes, similar to GLP-1 inhibitors. Because these peptides reduce weight and body fat and improve insulin resistance, they not only have excellent hypoglycemic effects but should also be particularly effective for a high proportion of diabetic patients who are overweight. Many overweight or obese individuals, although not medically diagnosed with diabetes, exhibit prediabetic symptoms such as impaired glucose tolerance and excessively high postprandial blood glucose; the peptides of this invention are also suitable for people with prediabetes. Summary of the Invention

[0010] In one aspect, the present invention relates to a pharmaceutical formulation comprising a peptide compound of formula (VII) or a pharmaceutically acceptable salt or solvate thereof.

[0011] Y-aib-EGTF-X11-SD-X1-S-X12-X2-L-X3-X4-EA-X5-X6-X13-F-X7-X8-WL-X9-AG-X10 (VII)

[0012] Wherein, X1 represents an amino acid selected from L or Y; X2 represents an amino acid selected from Q, A, aib, or Y; X3 represents an amino acid selected from D or E; X4 represents an amino acid selected from E or K; X5 represents an amino acid selected from V or A; X6 represents an amino acid selected from K, R, or Q; X7 represents an amino acid selected from I or V; X8 represents an amino acid selected from E, Q, N, or A; X9 represents an amino acid selected from I or L; X10 represents an amino acid that is absent or not present; X11 represents an amino acid selected from T or I; X12 represents an amino acid selected from I, S, or K; and X13 represents an amino acid selected from L, E, or D.

[0013] Optionally, one or two amino acids selected from S or containing amino or thiol groups in the side chain are added to the C-terminus of X10, and the carboxyl group of the C-terminal amino acid is optionally amidated to form a C-terminal amide, wherein the amino acid has the formula

[0014] (II) or (III),

[0015] The wavy line represents the connection point to the adjacent group, n1 is an integer from 1 to 7, and when II or III is a C-terminal amino acid, its carboxyl group is COOH or CONH2. Preferably, the amino acid containing a side chain amino group is lysine, and the amino acid containing a side chain thiol group is cysteine.

[0016] Optionally, the amino acid containing a side-chain amino group added to the C-terminus of X10 is modified with a long-acting group on its side chain amino group, preferably, the long-acting group has the structure of formula (IV):

[0017] O1-O2-O3-O4-O5-O6-O7-O8-(IV),

[0018] Where O1 represents the structure of formula (V) or (VI):

[0019] (V) or (VI)

[0020] Where n2 is an integer between 6 and 24, preferably between 10 and 24, and even more preferably between 16 and 22;

[0021] Wherein the wavy line represents the linking point of an amino group connected to an adjacent group, O2-O3-O4-O5-O6-O7-O8- represents a linker, wherein each of O2 to O8 is independently represented by any one of the following amino acid residues or long chain structures: α-Glu, γ-Glu, α-Asp, β-Asp, α-hGlu, δ-hGlu, Gly, Ala, β-Ala, GABA or PEG2, or one or more of O2 to O8 are absent, provided that at least two of O2 to O8 are present, preferably, O2 to O8 contain at least one negatively charged portion.

[0022] In one aspect, the present invention relates to a pharmaceutical preparation comprising a peptide compound of formula (I), or a salt or solvate thereof, and a pharmaceutically acceptable carrier or excipient.

[0023] Y-aib-EGTFTSD-X1-SI-X2-L-X3-X4-EA-X5-X6-LF-X7-X8-WL-X9-AG-X10 (I)

[0024] Wherein, X1 represents an amino acid selected from L or Y, X2 represents an amino acid selected from Q, A, aib or Y, X3 represents an amino acid selected from D or E, X4 represents an amino acid selected from E or K, X5 represents an amino acid selected from V or A, X6 represents an amino acid selected from K or R, X7 represents an amino acid selected from I or V, X8 represents an amino acid selected from E, A, Q or N, X9 represents an amino acid selected from I or L, and X10 represents an amino acid that is not present or GPSSGAPPP, GPPSGAPPP, GPSSGKPPP, GPSSGEPPP, GPSSaibAPPP, GPSSGAPP, GPSSGAP, GPSSGA, GPSSG, GPSS, GPS, GP, G.

[0025] Optionally, one or two amino acids selected from S or containing amino or thiol groups in the side chain are added to the C-terminus of X10, and the carboxyl group of the C-terminal amino acid is optionally amidated to form a C-terminal amide, wherein the amino acid has the formula

[0026] (II) or (III),

[0027] The wavy line represents the connection point to the adjacent group, and n1 is an integer from 1 to 7; when II or III is a C-terminal amino acid, its carboxyl part is COOH or CONH2;

[0028] Optionally, the amino acid containing a side-chain amino group added to the C-terminus of X10 is modified with a long-acting group on its side chain amino group, preferably, the long-acting group has the structure of formula (IV):

[0029] O1-O2-O3-O4-O5-O6-O7-O8-(IV),

[0030] Where O1 represents the structure of formula (V) or (VI):

[0031] (V) or (VI)

[0032] Where n2 is an integer between 6 and 24, preferably between 10 and 24, and even more preferably between 16 and 22;

[0033] Wherein the wavy line represents the linking point of an amino group connected to an adjacent group, O2-O3-O4-O5-O6-O7-O8- represents a linker, wherein each of O2 to O8 is independently represented by any one of the following amino acid residues or long chain structures: α-Glu, γ-Glu, α-Asp, β-Asp, α-hGlu, δ-hGlu, Gly, Ala, β-Ala, GABA or PEG2, or one or more of O2 to O8 are absent, provided that at least two of O2 to O8 are present, preferably, O2 to O8 contain at least one negatively charged portion.

[0034] According to any of the foregoing aspects of the pharmaceutical formulation, wherein O2-O3-O4-O5-O6-O7-O8- represents the linker selected from the group consisting of γGlu-PEG2-γGlu-, γGlu-PEG2-2×γGlu-, γGlu-PEG2-, γGlu-2×PEG2-, γGlu-3×PEG2-, γGlu-PEG2-γGlu-PEG2-, γGlu-2×PEG2-γGlu-, γGlu-2×PEG2-2×γGlu-, 2×γGlu-, 2×γGlu-PEG2-, 2×γGlu-PEG2-γGlu-, 2×γGlu-PEG2-γGlu-, 2×γGlu-2×PEG2-, 2×γGlu-2×PEG2-γGlu-, 2×γGlu-2×PEG2-2×γGlu-.

[0035] According to any of the foregoing aspects of the pharmaceutical formulation, in some embodiments, O2-O3-O4-O5-O6-O7-O8- represents the linker γGlu-PEG2-, γGlu-2×PEG2-, γGlu-3×PEG2- selected from the group consisting of, and O1 represents the structure of formula (V) or (VI).

[0036] According to any of the foregoing pharmaceutical formulations, in some embodiments, O2-O3-O4-O5-O6-O7-O8- represents the linker γGlu-PEG2-, γGlu-2×PEG2-, γGlu-3×PEG2- selected from the group consisting of, and O1 represents the structure of formula (V).

[0037] According to any of the foregoing pharmaceutical formulations, in some embodiments, O2-O3-O4-O5-O6-O7-O8- represents the linker γGlu-2×PEG2-, γGlu-3×PEG2- selected from the group consisting of, and O1 represents the structure of formula (V).

[0038] According to any of the foregoing pharmaceutical formulations, in some embodiments, O2-O3-O4-O5-O6-O7-O8- represents the linker γGlu-2×PEG2-, and O1 represents the structure of formula (V).

[0039] According to any of the foregoing pharmaceutical formulations, in some embodiments, O2-O3-O4-O5-O6-O7-O8- represents the linker γGlu-2×PEG2-, O1 represents the structure of formula (V), and n2 is an integer from 16 to 22;

[0040] Optionally, in any of the foregoing pharmaceutical formulations, the lysine side chain amino-conjugated long-acting group has a structure of formula (VIII) or formula (IX):

[0041] (VIII);

[0042] (IX);

[0043] The wavy line represents the connection point to the adjacent amino acid residue.

[0044] Optionally, the C-terminal amino acid containing a side-chain thiol group added to X10 has its side-chain thiol group modified by a long-acting group of formula (IV). Optionally, a reactive group capable of reacting with the thiol group to form a covalent bond can be added between the side-chain thiol group and the long-acting group of the C-terminal amino acid as needed.

[0045] In some embodiments, the linking relationship between the side chain thiol of the amino acid containing the side chain thiol and the long-acting group is: side chain thiol of the amino acid containing the side chain thiol - thiol reactive group - optional linking group L - long-acting group.

[0046] In some embodiments, the side chain thiol of the amino acid containing the side chain thiol is attached to one end of the linking group L after reacting with a Michael reaction acceptor (e.g., maleimide or vinyl sulfone) or a thiol reactive group (e.g., iodoacetic acid or bromoacetic acid). Preferably, the other end of the linking group L is further covalently attached to a long-acting group of formula IV.

[0047] In some embodiments, the linking group L is -(CH2). n3 - and -(CH2CH2O) n4 -A long chain formed by arranging and combining elements according to structural needs, linked together by covalent bonds; or -(CH2) n3 -、-(CH2CH2O) n4 - Optionally, one or both ends of the chain contain amino or carboxyl groups, forming a long chain linked together by amide bonds. For example, the linking group L is selected from -NH-(CH2). n5 -(CH2CH2O) n6 -(CH2) n7 -,-NH-(CH2) n5 -(CH2CH2O) n6 -(CH2) n7 -NH-, -NH-(CH2) n5 -(CH2CH2O) n6 -(CH2) n7 -CO-,-NH-(CH2) n5 -(CH2CH2O) n6 -(CH2) n7 -NHCO-(CH2) n8 -,-NH-(CH2) n5 -(CH2CH2O) n6 -(CH2) n7 -NHCO-(CH2) n8 -NH- or any combination thereof, where n3, n4, n5, n6, n7, and n8 are each integers from 0 to 10, for example, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.

[0048] In some implementations, L is -NH-CH2-(CH2CH2O)3-(CH2)3-NH-.

[0049] In some embodiments, non-limiting illustrative examples of the Michael reaction acceptor or thiol reactive group being linked to the linking group L include

[0050]

[0051] The structures of the Michael reaction acceptor or thiol reactive group reacting with the side chain thiol group of an amino acid containing a side chain thiol group, as exemplified above, are as follows:

[0052]

[0053] The wavy line indicates the connection point with a long-acting group of formula (IV), for example, connected to O8. * indicates the connection point between the side chain thiol group of an amino acid containing a side chain thiol group and the other part of the amino acid.

[0054] Optionally, any amino acid in the polypeptide fragment represented by X10 may be replaced by an amino acid with an amino or thiol group on its side chain, said amino acid having the structure of formula (II) or formula (III). Optionally, the amino acid containing a side chain amino group is modified with a long-acting group on its side chain amino group, preferably having the structure of formula (IV); optionally, the amino acid containing a side chain thiol group is modified with a long-acting group on its side chain thiol group, preferably having the structure of formula (IV); optionally, a reactive group capable of reacting with the thiol group to form a covalent bond may be added between the side chain thiol group and the long-acting group as needed.

[0055] Optionally, the polypeptide fragments GPPSGAPPP, GPSSGKPPP, GPSSGEPPP, and GPSSaibAPPP represented by X10 can be reduced by 1, 2, 3, 4, 5, 6, 7, or 8 amino acids from the C-terminus to the N-terminus.

[0056] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is Y. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is I. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is V.

[0057] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is Q. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is Y. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is A. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is Aib.

[0058] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X3 is D. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X3 is E.

[0059] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X4 is E. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X4 is K.

[0060] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X5 is V. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X5 is A.

[0061] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X6 is K. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X6 is R.

[0062] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X7 is I. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X7 is V.

[0063] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X8 is E. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X8 is N. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X8 is Q. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X8 is A.

[0064] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X9 is I. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X9 is Aib. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X9 is V.

[0065] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X10 is absent. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X10 is GPSSGAPPPSK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X10 is GPSSGAPPPSC. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X10 is GPSSGAPPPC. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X10 is GPSSGAPPP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X10 is GPSSGAPP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X10 is GPSSGAP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X10 is GPSS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X10 is GPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X10 is GP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X10 is G.

[0066] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X10 is GPSSGAPPPC. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X10 is GPSSGAPPP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X10 is GPSSGAPP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X10 is GPSSGAP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X10 is GPSSGA. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X10 is GPSSG. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X10 is GPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X10 is GP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X10 is G.

[0067] According to any of the foregoing pharmaceutical formulations, in some embodiments, X2 is Q and X10 is GPSSGAPPPS.

[0068] According to any of the foregoing pharmaceutical formulations, in some embodiments, X2 is Q and X10 is GPSSGAPPPK.

[0069] According to any of the foregoing pharmaceutical formulations, in some embodiments, X2 is Q and X10 is GPSSGAPPPC.

[0070] According to any of the foregoing pharmaceutical formulations, in some embodiments, X2 is A and X10 is GPSSGAPPPS.

[0071] According to any of the foregoing pharmaceutical formulations, in some embodiments, X2 is A and X10 is GPSSGAPPPK.

[0072] According to any of the foregoing pharmaceutical formulations, in some embodiments, X2 is A and X10 is GPSSGAPPPC.

[0073] According to any of the foregoing pharmaceutical formulations, in some embodiments, X2 is aib and X10 is GPSSGAPPPS.

[0074] According to any of the foregoing pharmaceutical formulations, in some embodiments, X2 is aib and X10 is GPSSGAPPPK.

[0075] According to any of the foregoing pharmaceutical formulations, in some embodiments, X2 is aib and X10 is GPSSGAPPPC.

[0076] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is Q and X10 is GPSSGAPPP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is Q and X10 is GPSSGAPP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is Q and X10 is GPSSGAP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is Q and X10 is GPSSGA. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is Q and X10 is GPSSG. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is Q and X10 is GPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is Q and X10 is GP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is Q and X10 is G.

[0077] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is A and X10 is GPSSGAPPP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is A and X10 is GPSSGAPP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is A and X10 is GPSSGAP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is A and X10 is GPSSGA. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is A and X10 is GPSSG. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is A and X10 is GPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is A and X10 is GP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is A and X10 is G.

[0078] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is aib and X10 is GPSSGAPPP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is aib and X10 is GPSSGAPP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is aib and X10 is GPSSGAP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is aib and X10 is GPSSGA. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is aib and X10 is GPSSG. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is aib and X10 is GPSS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is aib and X10 is GPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is aib and X10 is GP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is aib and X10 is G.

[0079] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X2 is Q. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X2 is A. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X2 is aib.

[0080] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, and X10 is GPSSGAPPPC. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, and X10 is GPSSGAPPP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, and X10 is GPSSGAPP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, and X10 is GPSSGAP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, and X10 is GPSSGA. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, and X10 is GPSSG. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, and X10 is GPSS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, and X10 is GPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, and X10 is GP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, and X10 is G.

[0081] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, and X10 is GPSSGAPPPC. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, and X10 is GPSSGGAPPP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, and X10 is GPSSGAP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, and X10 is GPSSGA. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, and X10 is GPSSG. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, and X10 is GPSS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, and X10 is GPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, and X10 is GP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, and X10 is G.

[0082] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, and X10 is GPSSGAPPPC. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, and X10 is GPSSGAPPP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, and X10 is GPSSGAPP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, and X10 is GPSSGAP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, and X10 is GPSSGA. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, and X10 is GPSSG. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, and X10 is GPSS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, and X10 is GPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, and X10 is GP. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, and X10 is G.

[0083] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X5 is V, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X5 is V, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X5 is V, and X10 is GPSSGAPPPC.

[0084] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X5 is V, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X5 is V, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X5 is V, and X10 is GPSSGAPPPC.

[0085] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X5 is V, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X5 is V, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X5 is V, and X10 is GPSSGAPPPC.

[0086] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X5 is V, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X5 is V, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X5 is V, and X10 is GPSSGAPPPC.

[0087] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X5 is V, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X5 is V, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X5 is V, and X10 is GPSSGAPPPC.

[0088] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X5 is V, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X5 is V, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X5 is V, and X10 is GPSSGAPPPC.

[0089] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, and X10 is GPSSGAPPPC.

[0090] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, and X10 is GPSSGAPPPC.

[0091] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, and X10 is GPSSGAPPPC.

[0092] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, X7 is I, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, X7 is I, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, X7 is I, and X10 is GPSSGAPPPC.

[0093] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X7 is I, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X7 is I, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X7 is I, and X10 is GPSSGAPPPC.

[0094] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, X7 is I, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, X7 is I, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, X7 is I, and X10 is GPSSGAPPPC.

[0095] According to any of the foregoing pharmaceutical formulations, in some embodiments, X2 is A and X5 is V.

[0096] According to any of the foregoing pharmaceutical formulations, in some embodiments, X2 is aib and X5 is V.

[0097] According to any of the foregoing pharmaceutical formulations, in some embodiments, X1 is L, X2 is A, and X5 is V.

[0098] According to any of the foregoing pharmaceutical formulations, in some embodiments, X1 is L, X2 is aib, and X5 is V.

[0099] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, and X5 is V. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, and X5 is V.

[0100] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, and X5 is V. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, and X5 is V.

[0101] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, and X7 is I. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, and X7 is I.

[0102] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is Q and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is Q, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is Q, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is Q, X9 is L, and X10 is GPSSGAPPPC.

[0103] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is A and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is A, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is A, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is A, X9 is L, and X10 is GPSSGAPPPC.

[0104] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is aib and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is aib, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is aib, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X2 is aib, X9 is L, and X10 is GPSSGAPPPC.

[0105] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X9 is L, and X10 is GPSSGAPPPC.

[0106] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X9 is L, and X10 is GPSSGAPPPC.

[0107] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X9 is L, and X10 is GPSSGAPPPC.

[0108] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X5 is V, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X5 is V, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X5 is V, X9 is L, and X10 is GPSSGAPPPC.

[0109] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X5 is V, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X5 is V, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X5 is V, X9 is L, and X10 is GPSSGAPPPC.

[0110] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X5 is V, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X5 is V, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X5 is V, X9 is L, and X10 is GPSSGAPPPC.

[0111] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X5 is V, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X5 is V, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X5 is V, X9 is L, and X10 is GPSSGAPPPC.

[0112] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X5 is V, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X5 is V, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X5 is V, X9 is L, and X10 is GPSSGAPPPC.

[0113] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X5 is V, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X5 is V, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X5 is V, X9 is L, and X10 is GPSSGAPPPC.

[0114] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, X9 is L, and X10 is GPSSGAPPPC.

[0115] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X9 is L, and X10 is GPSSGAPPPC.

[0116] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, X9 is L, and X10 is GPSSGAPPPC.

[0117] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, X7 is I, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, X7 is I, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, X7 is I, X9 is L, and X10 is GPSSGAPPPC.

[0118] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X7 is I, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X7 is I, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X7 is I, X9 is L, and X10 is GPSSGAPPPC.

[0119] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, X7 is I, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, X7 is I, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, X7 is I, X9 is L, and X10 is GPSSGAPPPC.

[0120] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, X7 is V, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, X7 is V, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, X7 is V, X9 is L, and X10 is GPSSGAPPPC.

[0121] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X7 is V, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X7 is V, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X7 is V, X9 is L, and X10 is GPSSGAPPPC.

[0122] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, X7 is I, X9 is L, and X10 is GPSSGAPPPS. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, X7 is V, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, X7 is V, X9 is L, and X10 is GPSSGAPPPC.

[0123] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X3 is E and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X3 is E, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X3 is E, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X3 is E, X4 is K, X5 is V, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X3 is E, X4 is K, X5 is V, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X3 is E, X4 is K, X5 is V, X7 is I, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X3 is E, X4 is K, X5 is V, X7 is I, X8 is E, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X3 is E, X4 is K, X5 is V, X6 is R, X7 is I, X8 is E, X9 is L, and X10 is GPSSGAPPPK. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X3 is E, X4 is K, X5 is V, X6 is R, X7 is I, X8 is E, X9 is L, and X10 is GPSSGAPPPK, wherein the C-terminal lysine side chain amino-conjugated long-acting group of X10. Preferably, the C-terminal lysine side chain amino-conjugated long-acting group of X10 has a structure of formula (VIII) or formula (IX). In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A or aib, X3 is E, X4 is K, X5 is V, X6 is R, X7 is I, X8 is E, X9 is L, and X10 is GPSSGAPPPK. Preferably, the C-terminal lysine side chain amino-conjugated long-acting group of X10. More preferably, the C-terminal lysine side chain amino-conjugated long-acting group of X10 has the structure of formula (VIII) or formula (IX). In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X6 is R, X7 is I, X8 is E, X9 is L, and X10 is GPSSGAPPPK. Preferably, the C-terminal lysine side chain amino-conjugated long-acting group of X10.

[0124] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is Y and X2 is Q. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is Y and X2 is A. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is Y and X2 is Aib. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is Y and X2 is Y.

[0125] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X4 is K and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is Y, X4 is K, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is Y, X2 is Y, X4 is K, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is Y, X2 is Y, X3 is E, X4 is K, X5 is V, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is Y, X2 is Y, X3 is E, X4 is K, X5 is V, X7 is V, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is Y, X2 is Y, X3 is E, X4 is K, X5 is V, X7 is V, X8 is N, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is Y, X2 is Y, X3 is E, X4 is K, X5 is V, X7 is V, X8 is N, and X9 is I. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is Y, X2 is Y, X3 is E, X4 is K, X5 is V, X7 is I, X8 is E, and X9 is L.

[0126] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X9 is I. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X5 is V, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X5 is V, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Q, X3 is E, X4 is K, X5 is V, X7 is I, X8 is E, and X9 is L.

[0127] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X2 is A. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X5 is V, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X5 is V, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X7 is I, X8 is E, and X9 is L.

[0128] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X2 is aib. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X5 is V, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X5 is V, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, and X9 is L. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, X7 is I, X8 is E, and X9 is L.

[0129] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L and X2 is Y. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Y, and X5 is V. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Y, X3 is E, and X5 is V. In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, X1 is L, X2 is Y, X3 is E, X4 is K, and X5 is V.

[0130] According to any of the foregoing pharmaceutical formulations, wherein the peptide compound is selected from:

[0131] Compound 1. Y(aib)EGTFTSDYSIYLDEEAVRLFVNWLIAGGPSSGAPPPS-NH2

[0132] Compound 2. Y(aib)EGTFTSDYSIYLDEEAVKLFVNWLIAGGPSSGAPPPS-NH2

[0133] Compound 3. Y(aib)EGTFTSDYSIYLDEEAVKLFVQWLIAGGPSSGAPPPS-NH2

[0134] Compound 4.

[0135]

[0136] Compound 5. Y(aib)EGTFTSDYSIYLDEEAVRLFVNWLIAGGPSSGAPPPSC-NH2

[0137] Compound 6.

[0138]

[0139] Compound 7. Y(aib)EGTFTSDYSIYLDEEAVKLFVNWLIAG-NH2

[0140] Compound 8. Y(aib)EGTFTSDYSIYLEKEAVRLFVNWLIAGGPSSGAPPPS-NH2

[0141] Compound 9. Y(aib)EGTFTSDYSIYLEKEAVKLFVNWLIAGGPSSGAPPPS-NH2

[0142] Compound 10. Y(aib)EGTFTSDYSIYLEKEAAKLFVNWLIAGGPSSGAPPPS-NH2

[0143] Compound 11.

[0144]

[0145] Compound 12. Y(aib)EGTFTSDYSIYLEKEAVKLFVNWLIAGGPSSGAPPPK-NH2

[0146] Compound 13.

[0147]

[0148] Compound 14.

[0149]

[0150] Compound 15. Y(aib)EGTFTSDYSIYLEKEAVRLFVNWLLAG-NH2

[0151] Compound 16. Y(aib)EGTFTSDYSIYLEKEAVRLFIEWLIAGGPSSGAPPPS-NH2

[0152] Compound 17.

[0153]

[0154] Compound 18. Y(aib)EGTFTSDLSIQLEKEAARLFIEWLLAGGPSSGAPPPS-NH2

[0155] Compound 19. Y(aib)EGTFTSDLSIQLEKEAVRLFIEWLLAGGPSSGAPPPS-NH2

[0156] Compound 20.

[0157]

[0158] Compound 21.

[0159]

[0160] Compound 22.

[0161]

[0162] Compound 23.

[0163]

[0164] Compound 24. Y(aib)EGTFTSDLSIQLEKEAVKLFIEWLLAGGPSSGAPPPS-NH2

[0165] Compound 25. Y(aib)EGTFTSDLSIQLEKEAVRLFIEWLLAG-NH2

[0166] Compound 26.

[0167]

[0168] Compound 27.

[0169]

[0170] Compound 28.

[0171]

[0172] compound 29

[0173]

[0174] compound 30

[0175]

[0176] Compound 31. Y(aib)EGTFTSDLSIALEKEAVRLFIEWLLAGGPSSGAPPPS-NH2

[0177] Compound 32. Y(aib)EGTFTSDLSIALEKEAVRLFIEWLLAGGPSSGAPPPSC-NH2

[0178] Compound 33. Y(aib)EGTFTSDLSIALEKEAVRLFIEWLLAGGPSSGAPPPC-NH2

[0179] Compound 34. Y(aib)EGTFTSDLSIALEKEAVRLFIEWLLAGGPSSGAPPPSK-NH2

[0180] Compound 35. Y(aib)EGTFTSDLSIALEKEAVRLFIEWLLAGGPSSGAPPPK-NH2

[0181] Compound 36. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIEWLLAGGPSSGAPPPS-NH2

[0182] Compound 37. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIEWLLAGGPSSGAPPPSC-NH2

[0183] Compound 38. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIEWLLAGGPSSGAPPPC-NH2

[0184] Compound 39. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIEWLLAGGPSSGAPPPSK-NH2

[0185] Compound 40. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIEWLLAGGPSSGAPPPK-NH2

[0186] Compound 41. Y(aib)EGTFTSDLSIALEKEAVRLFIEWLLAG-NH2

[0187] Compound 42. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIEWLLAG-NH2

[0188] Compound 43. Y(aib)EGTFTSDLSIALEKEAVKLFIEWLLAGGPSSGAPPPS-NH2

[0189] Compound 44. Y(aib)EGTFTSDLSIALEKEAVKLFIEWLLAGGPSSGAPPPK-NH2

[0190] Compound 45. Y(aib)EGTFTSDLSIALEKEAVKLFIEWLLAGGPSSGAPPPSK-NH2

[0191] Compound 46. Y(aib)EGTFTSDLSIALEKEAVKLFIEWLLAGGPSSGAPPPSC-NH2

[0192] Compound 47. Y(aib)EGTFTSDLSIALEKEAVKLFIEWLLAGGPSSGAPPPC-NH2

[0193] Compound 48. Y(aib)EGTFTSDLSI(aib)LEKEAVKLFIEWLLAGGPSSGAPPPS-NH2

[0194] Compound 49. Y(aib)EGTFTSDLSI(aib)LEKEAVKLFIEWLLAGGPSSGAPPPK-NH2

[0195] Compound 50. Y(aib)EGTFTSDLSI(aib)LEKEAVKLFIEWLLAGGPSSGAPPPSK-NH2

[0196] Compound 51. Y(aib)EGTFTSDLSI(aib)LEKEAVKLFIEWLLAGGPSSGAPPPC-NH2

[0197] Compound 52. Y(aib)EGTFTSDLSI(aib)LEKEAVKLFIEWLLAGGPSSGAPPPSC-NH2

[0198] Compound 53. Y(aib)EGTFTSDLSIQLEKEAVRLFVNWLLAGGPSSGAPPPS-NH2

[0199] Compound 54. Y(aib)EGFTSDLSIALEKEAVRLFVNWLLAGGPSSGAPPPK-NH2

[0200] Compound 55. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFVNWLLAGGPSSGAPPPC-NH2

[0201] Compound 56. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFVNWLLAGGPSSGAPPPK-NH2

[0202] Compound 57. Y(aib)EGTFTSDYSIYLDEEAVRLFIEWLIAGGPSSGAPPPK-NH2

[0203] Compound 58. Y(aib)EGTFTSDLSIQLEKEAVRLFVNWLLAGGPSSGAPPPS-NH2

[0204] Compound 59. Y(aib)EGTFTSDLSIALEKEAVRLFVNWLLAGGPSSGAPPPS-NH2

[0205] Compound 60. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFVNWLLAGGPSSGAPPPS-NH2

[0206] Compound 61. Y(aib)EGTFTSDYSIYLDEEAVRLFIEWLIAGGPSSGAPPPS-NH2

[0207] Compound 62 Y(aib)EGTFTSDLSIQLEKEAVRLFIEWLLAGGPSSGAPPPSC-NH2

[0208] Compound 63 Y(aib)EGTFTSDLSIQLEKEAVRLFIEWLLAGGPSSGAPPPC-NH2

[0209] Compound 64 Y(aib)EGTFTSDLSIQLEKEAVRLFIEWLLAGGPSSGAPPPSK-NH2

[0210] Compound 65 Y(aib)EGTFTSDLSIQLEKEAVRLFIEWLLAGGPSSGAPPPK-NH2

[0211] Compound 66 Y(aib)EGTFTSDLSIQLEKEAVKLFIEWLLAGGPSSGAPPPSC-NH2

[0212] Compound 67 Y(aib)EGTFTSDLSIQLEKEAVKLFIEWLLAGGPSSGAPPPC-NH2

[0213] Compound 68 Y(aib)EGTFTSDLSIQLEKEAVKLFIEWLLAGGPSSGAPPPSK-NH2

[0214] Compound 69 Y(aib)EGTFTSDLSIQLEKEAVKLFIEWLLAGGPSSGAPPPK-NH2

[0215] Compound 70. Y(aib)EGTFTSDYSIYLDEEAVRLFVNWLIAGGPSSGAPPPSK-NH2

[0216] Compound 71. Y(aib)EGTFTSDYSIYLDEEAVKLFVNWLIAGGPSSGAPPPSK-NH2

[0217] Compound 72. Y(aib)EGTFTSDYSIYLDEEAVRLFVNWLIAGGPSSGAPPPK-NH2

[0218] Compound 73. Y(aib)EGTFTSDYSIYLDEEAVKLFVNWLIAGGPSSGAPPPK-NH2

[0219] Compound 74. Y(aib)EGTFTSDYSIYLDEEAVKLFVNWLIAGGPSSGAPPPSC-NH2

[0220] Compound 75. Y(aib)EGTFTSDYSIYLDEEAVRLFVNWLIAGGPSSGAPPPC-NH2

[0221] Compound 76. Y(aib)EGTFTSDYSIYLDEEAVKLFVNWLIAGGPSSGAPPPC-NH2

[0222] Compound 77. Y(aib)EGTFTSDYSIYLEKEAVRLFVNWLIAGGPSSGAPPPSK-NH2

[0223] Compound 78. Y(aib)EGTFTSDYSIYLEKEAVKLFVNWLIAGGPSSGAPPPSK-NH2

[0224] Compound 79. Y(aib)EGTFTSDYSIYLEKEAVRLFVNWLIAGGPSSGAPPPK-NH2

[0225] Compound 80. Y(aib)EGTFTSDYSIYLEKEAVRLFVNWLIAGGPSSGAPPPSC-NH2

[0226] Compound 81. Y(aib)EGTFTSDYSIYLEKEAVKLFVNWLIAGGPSSGAPPPSC-NH2

[0227] Compound 82. Y(aib)EGTFTSDYSIYLEKEAVRLFVNWLIAGGPSSGAPPPC-NH2

[0228] Compound 83. Y(aib)EGTFTSDYSIYLEKEAVKLFVNWLIAGGPSSGAPPPC-NH2

[0229] Compound 84. Y(aib)EGTFTSDYSIYLEKEAVRLFIEWLIAGGPSSGAPPPSK-NH2

[0230] Compound 85. Y(aib)EGTFTSDYSIYLEKEAVRLFIEWLIAGGPSSGAPPPK-NH2

[0231] Compound 86. Y(aib)EGTFTSDYSIYLEKEAVRLFIEWLIAGGPSSGAPPPSC-NH2

[0232] Compound 87. Y(aib)EGTFTSDYSIYLEKEAVRLFIEWLIAGGPSSGAPPPC-NH2

[0233] Compound 88. Y(aib)EGTFTSDYSEYLEKEAVRLFIEWLIAG-NH2

[0234] Compound 89. Y(aib)EGTFTSDLSIQLEKEAVRLFVNWLLAG-NH2

[0235] Compound 90. Y(aib)EGTFTSDLSIALEKEAVRLFVNWLLAG-NH2

[0236] Compound 91. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFVNWLLAG-NH2

[0237] Compound 92. Y(aib)EGTFTSDLSIQLEKEAVRLFIEWLLAGGPSSGA-NH2

[0238] Compound 93. Y(aib)EGTFTSDYSIYLDEEAVRLFIEWLIAGGPSSGAPPPS-NH2

[0239] Compound 94. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIEWLLAGGPSSGAP-NH2

[0240] Compound 95. Y(aib)EGTFTSDYSIYLEKEAVRLFIEWLIAGGPSSGAPP-NH2

[0241] Compound 96. Y(aib)EGTFTSDYSIYLDEEAVRLFVNWLIAGGPSSGAPP-NH2

[0242] Compound 97. Y(aib)EGTFTSDYSIYLEKEAVKLFVNWLIAGGPSSGAP-NH2

[0243] Compound 98. Y(aib)EGTFTSDYSIYLEKEAVRLFIAWLLAGPGPSSGAPPPS-NH2

[0244] Compound 99. Y(aib)EGTFTSDYSIYLDEEAVRLFVNWLIAGG-NH2

[0245] Compound 100. Y(aib)EGTFTSDYSIYLEKEAVRLFIEWLIAGGPSSGAP-NH2

[0246] Compound 101. Y(aib)EGTFTSDLSKALEKEAVRLFIEWLLAGGPSSGAPPPS-NH2

[0247] Compound 102. Y(aib)EGTFTSDYSIYLEKEAVKLFIEWLIAGGPSSGAPPPS-NH2

[0248] Compound 103. Y(aib)EGTFTSDYSIYLDEEAVRLFVNWLIAGGPSSGA-NH2

[0249] Compound 104. Y(aib)EGTFTSDLSIQLEKEAVKEFIAWLIAGGPSSGAPPPS-NH2

[0250] Compound 105. Y(aib)EGTFTSDYSIYLDEEAVRLFIAWLIAGGPSSGAPPPS-NH2

[0251] Compound 106. Y(aib)EGTFTSDYSIYLEKEAVKLFVNWLIAGG-NH2

[0252] Compound 107. Y(aib)EGTFTSDLSIALEKEAVRLFVNWLIAGGPSSGAPPPS-NH2

[0253] Compound 108. Y(aib)EGTFTSDYSIYLDEEAVRLFVNWLIAGGPSSGAP-NH2

[0254] Compound 109. Y(aib)EGTFTSDYSIYLEKEAVKLFVNWLIAGGPSSG-NH2

[0255] Compound 110. Y(aib)EGTFTSDYSIYLEKEAVRLFIEWLIAGGPSSGA-NH2

[0256] Compound 111. Y(aib)EGTFTSDLSIALEKEAVRLFIEWLLAGGPSSGA-NH2

[0257] Compound 112. Y(aib)EGTFTSDYSIYLEKEAVKLFVNWLIAGGP-NH2

[0258] Compound 113. Y(aib)EGTFTSDYSIYLDEEAVRLFVNWLLAGGPSSGAPPP-NH2

[0259] Compound 114. Y(aib)EGTFTSDLSIQLEKEAVRLFVNWLIAGGPSSGAPPPS-NH2

[0260] Compound 115. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIEWLLAGGPSSGA-NH2

[0261] Compound 116. Y(aib)EGTFTSDYSIYLEKEAVRLFIEWLIAGGPSS-NH2

[0262] Compound 117. Y(aib)EGTFTSDLSIQLEKEAVRLFIAWLIAGGPSSGAPPPS-NH2

[0263] Compound 118. Y(aib)EGTFTSDYSIYLEKEAVKLFVNWLLAGGPSSGAPPP-NH2

[0264] Compound 119. Y(aib)EGTFTSDLSIALEKEAVRLFIEWLLAGGPS-NH2

[0265] Compound 120. Y(aib)EGTFTSDYSIYLDEEAVRLFVNWLIAGGPSS-NH2

[0266] Compound 121. Y(aib)EGTFTSDLSIQLEKEAVRLFIEWLIAGGPSSGAPPP-NH2

[0267] Compound 122. Y(aib)EGTFTSDYSIYLDEEAVRLFVNWLIAGGP-NH2

[0268] Compound 123. Y(aib)EGTFTSDYSIYLEKEAVRLFIEWLLAGGPSSGAPPP-NH2

[0269] Compound 124. Y(aib)EGTFTSDLSIQLEKEAVRLFIEWLLAGG-NH2

[0270] Compound 125. Y(aib)EGTFTSDLSIQLEKEAVKLFIEWLIAGGPSSGAPPP-NH2

[0271] Compound 126. Y(aib)EGTFTSDYSIYLEKEAVKLFIAWLIAGGPSSGAPPPS-NH2

[0272] Compound 127. Y(aib)EGTFTSDLSIALEKEAVRLFIAWLIAGGPSSGAPPPS-NH2

[0273] Compound 128. Y(aib)EGTFTSDLSIQLEKEAVKLFIEWLLAGGPSSGAPP-NH2

[0274] Compound 129. Y(aib)EGTFTSDYSIYLEKEAVKEFIAWLLAGGPSSGAPPP-NH2

[0275] Compound 130. Y(aib)EGTFTSDLSIQLEKEAVRLFIEWLLAGGP-NH2

[0276] Compound 131. Y(aib)EGTFTSDLSIQLEKEAVKLFIEWLLAGGPSSGAP-NH2

[0277] Compound 132. Y(aib)EGTFTSDYSIYLEKEAVKLFVNWLIAGGPSSGA-NH2

[0278] Compound 133. Y(aib)EGTFISDLSIALEKEAVRLFIEWLLAGGPSSGAPPPS-NH2

[0279] Compound 134. Y(aib)EGTFTSDYSIYLEKEAVRLFIEWLIAGGP-NH2

[0280] Compound 135. Y(aib)EGTFTSDYSIYLEKEAVKLFVNWLIAGGPSS-NH2

[0281] Compound 136. Y(aib)EGTFTSDLSI(aib)LEKEAVKEFIAWLLAGGPSSGAPPPS-NH2

[0282] Compound 137. Y(aib)EGTFTSDLSIQLEKEAVKLFIEWLLAGGPSSGA-NH2

[0283] Compound 138. Y(aib)EGTFTSDLSIALEKEAVRLFIEWLLAGGPSSG-NH2

[0284] Compound 139. Y(aib)EGTFTSDYSIYLDEEAVRLFVNWLIAGGPS-NH2

[0285] Compound 140. Y(aib)EGTFTSDLSIQLEKEAVRLFIEWLLAGGPSS-NH2

[0286] Compound 141. Y(aib)EGTFTSDYSIYLEKEAVKLFVNWLIAGGPSSGAPP-NH2

[0287] Compound 142. Y(aib)EGTFTSDLSIALEKEAVRLFIEWLLAGGPSSGAP-NH2

[0288] Compound 143. Y(aib)EGTFTSDLSIQLEKEAVKLFIEWLLAGGPSSG-NH2

[0289] Compound 144. Y(aib)EGTFTSDYSIALEKEAVRLFIEWLLAGGPSSGAPPPS-NH2

[0290] Compound 145. Y(aib)EGTFTSDLSIQLEKEAVKLFIEWLLAGGPSS-NH2

[0291] Compound 146. Y(aib)EGTFTSDLSIQLEKEAVRLFIEWLLAGGPSSG-NH2

[0292] Compound 147. Y(aib)EGTFISDLSI(aib)LEKEAVRLFIEWLLAGGPSSGAPPPS-NH2

[0293] Compound 148. Y(aib)EGTFTSDYSIYLEKEAVRLFIEWLIAGGPSSG-NH2

[0294] Compound 149. Y(aib)EGTFTSDYSIYLEKEAVKEFIAWLIAGGPSSGAPPPS-NH2

[0295] Compound 150. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIEWLLAGGPSSGAPP-NH2

[0296] Compound 151. Y(aib)EGTFTSDLSIYLDEEAVRLFVNWLLAGGPSSGAPPPS-NH2

[0297] Compound 152. Y(aib)EGTFTSDLSIALEKEAVRLFIEWLLAGGP-NH2

[0298] Compound 153. Y(aib)EGTFTSDLSIQLEKEAVKLFIEWLLAGGPS-NH2

[0299] Compound 154. Y(aib)EGTFTSDLSIQLEKEAVRLFIEWLLAGGPSSGAPP-NH2

[0300] Compound 155. Y(aib)EGTFTSDYSIYLEKEAVRLFIEWLIAGG-NH2

[0301] Compound 156. Y(aib)EGTFTSDLSIQLEKEAVKLFIEWLLAGGP-NH2

[0302] Compound 157. Y(aib)EGFTSDLSIALEKEAVQDFVNWLIAGGPSSGAPPPS-NH2

[0303] Compound 158. Y(aib)EGTFTSDLSIQLEKEAVRLFIEWLLAGGPS-NH2

[0304] Compound 159. Y(aib)EGTFTSDLSIYLDEEAVRLFVNWLLAGGPSSGAP-NH2

[0305] Compound 160. Y(aib)EGTFTSDLSIALEKEAVRLFIEWLLAGGPSSGAPPP-NH2

[0306] Compound 161. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIAWLLAGGPSSGAPPPS-NH2

[0307] Compound 162. Y(aib)EGTFTSDYSIQLDEEAVRLFVNWLLAGGPSSGAPPPS-NH2

[0308] Compound 163. Y(aib)EGTFTSDLSIALEKEAVKEFIAWLIAGGPSSGAPPPS-NH2

[0309] Compound 164. Y(aib)EGTFTSDYSIYLEKEAVRLFIEWLIAGGPS-NH2

[0310] Compound 165. Y(aib)EGTFTSDYSIYLDEEAVRLFVNWLIAGGPSSG-NH2

[0311] Compound 166. Y(aib)EGTFTSDLSI(aib)LEKEAVQDFVNWLLAGGPSSGAPPPS-NH2

[0312] Compound 167. Y(aib)EGTFTSDLSIYLEKEAVKLFVNWLLAGGPSSGAP-NH2

[0313] Compound 168. Y(aib)EGTFTSDLSK(aib)LEKEAVRLFIEWLLAGGPSSGAPPP-NH2

[0314] Compound 169. Y(aib)EGTFTSDYSIYLDEEAVKEFIAWLIAGGPSSGAPPPS-NH2

[0315] Compound 170. Y(aib)EGTFTSDLSIYLEKEAVRLFIEWLLAGGPSSGAPP-NH2

[0316] Compound 171. Y(aib)EGTFTSDYSIALDEEAVRLFVNWLLAGGPSSGAPPPS-NH2

[0317] Compound 172. Y(aib)EGTFTSDLSIALEKEAVKLFVNWLLAGGPSSGAPPPS-NH2

[0318] Compound 173. Y(aib)EGTFTSDLSIYLEKEAVRLFIEWLLAGGPSSGAP-NH2

[0319] Compound 174. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIEWLLAGG-NH2

[0320] Compound 175. Y(aib)EGTFTSDLSIYLEKEAVKLFVNWLLAGGPSSGA-NH2

[0321] Compound 176. Y(aib)EGTFTSDLSIYLDEEAVRLFVNWLLAGGPS-NH2

[0322] Compound 177. Y(aib)EGTFTSDLSIYLDEEAVRLFVNWLLAGGPSSGAPPP-NH2

[0323] Compound 178. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIEWLLAGGPS-NH2

[0324] Compound 179. Y(aib)EGTFTSDLSIALEKEAVRLFIEWLLAGGPSSGAPP-NH2

[0325] Compound 180. Y(aib)EGTFTSDLSIYLEKEAVKLFVNWLLAGGPSSG-NH2

[0326] Compound 181. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIEWLLAGGPSSG-NH2

[0327] Compound 182. Y(aib)EGTFTSDYSIQLEKEAVRLFIEWLLAGGPSSGAPPPS-NH2

[0328] Compound 183. Y(aib)EGTFTSDLSIYLEKEAVKLFVNWLLAGGPSSGAPPP-NH2

[0329] Compound 184. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIEWLLAGGPSS-NH2

[0330] Compound 185. Y(aib)EGTFTSDLSIYLDEEAVRLFVNWLLAGGPSSGA-NH2

[0331] Compound 186. Y(aib)EGTFTSDYSI(aib)LEKEAVRLFIEWLLAGGPSSGAPPPS-NH2

[0332] Compound 187. Y(aib)EGTFTSDLSIALEKEAVRLFIEWLLAGG-NH2

[0333] Compound 188. Y(aib)EGTFTSDLSIQLEKEAVRLFIEWLLAGGPSSGAP-NH2

[0334] Compound 189. Y(aib)EGTFTSDYSIYLEKEAVKLFVNWLIAGGPS-NH2

[0335] Compound 190. Y(aib)EGTFTSDLSIYLEKEAVRLFIEWLLAGGPSSGAPPPS-NH2

[0336] Compound 191. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFVNWLLAGGPSSGAPPPS-NH2

[0337] Compound 192. Y(aib)EGTFTSDLSIYLDEEAVRLFVNWLLAGGPSSG-NH2

[0338] Compound 193. Y(aib)EGTFTSDLSIYLEKEAVKLFVNWLLAGGPS-NH2

[0339] Compound 194. Y(aib)EGTFTSDLSIYLDEEAVRLFVNWLLAGGPSS-NH2

[0340] Compound 195. Y(aib)EGTFTSDLSK(aib)LEKEAVRLFIEWLLAGGPSSGAPPPS-NH2

[0341] Compound 196. Y(aib)EGTFTSDLSIYLEKEAVRLFIEWLLAGGPSSG-NH2

[0342] Compound 197. Y(aib)EGTFTSDLSIYLEKEAVKLFVNWLLAGGPSSGAPP-NH2

[0343] Compound 198. Y(aib)EGTFTSDLSIYLEKEAVRLFIEWLLAGGPS-NH2

[0344] Compound 199. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIEWLLAGGP-NH2

[0345] Compound 200. Y(aib)EGTFTSDLSIYLEKEAVRLFIEWLLAGGP-NH2

[0346] Compound 201. Y(aib)EGTFTSDLSIYLDEEAVRLFVNWLLAGG-NH2

[0347] Compound 202. Y(aib)EGTFTSDLSIYLEKEAVKLFVNWLLAGGPSSGAPPPS-NH2

[0348] Compound 203. Y(aib)EGTFTSDLSIYLEKEAVRLFIEWLLAGGPSS-NH2

[0349] Compound 204. Y(aib)EGTFTSDLSIYLDEEAVRLFVNWLLAGGP-NH2

[0350] Compound 205. Y(aib)EGTFTSDLSIYLEKEAVKLFVNWLLAGGPSS-NH2

[0351] Compound 206. Y(aib)EGTFTSDLSIQLEKEAVKLFIEWLLAGG-NH2

[0352] Compound 207. Y(aib)EGTFTSDYSIYLEKEAVRLFIEWLLAGGPSSGAPPP-NH2

[0353] Compound 208. Y(aib)EGTFTSDLSIYLEKEAVKLFVNWLLAGGP-NH2

[0354] Compound 209. Y(aib)EGTFTSDLSIYLEKEAVRLFIEWLLAGGPSSGAPPP-NH2

[0355] Compound 210. Y(aib)EGTFTSDLSI(aib)LEKEAVRLFIEWLLAGGPSSGAPPP-NH2

[0356] Compound 211. Y(aib)EGTFTSDLSIYLEKEAVRLFIEWLLAGGPSSGA-NH2

[0357] Compound 212. Y(aib)EGTFTSDLSIYLEKEAVKLFVNWLLAGG-NH2

[0358] Compound 213. Y(aib)EGTFTSDYSI(aib)LDEEAVRLFVNWLLAGGPSSGAPPPS-NH2

[0359] Compound 214. Y(aib)EGTFTSDLSIALEKEAVRLFIEWLLAGGPSS-NH2

[0360] Compound 215. Y(aib)EGTFTSDLSIYLDEEAVRLFVNWLLAGGPSSGAPP-NH2

[0361] Compound 216. Y(aib)EGTFTSDLSI(aib)LEKEAVKLFVNWLLAGGPSSGAPPPS-NH2

[0362] Compound 217. Y(aib)EGTFTSDLSALEKEAVRLFIEWLLAGPGPSSGAPPPK-NH2

[0363] Compound 218. Y(aib)EGTFTSDLSIYLEKEAVRLFIEWLLAGG-NH2

[0364] Compound 219. Y(aib)EGTFTSDLSIQLEKEAVKLFVNWLLAGGPSSGAPPPS-NH2

[0365] Compound 220.

[0366]

[0367] Compound 221

[0368] .

[0369] In any of the foregoing pharmaceutical formulations, the peptide compound contains only natural amino acids.

[0370] Pharmaceutical formulations can be made into, for example, tablets, coated tablets, sugar lozenges, hard and soft gelatin capsules, solutions, emulsions, or suspensions for oral administration. However, they can also be formulated as suppositories for rectal administration or as injections for non-gastrointestinal administration. Commonly used non-gastrointestinal routes of administration include subcutaneous injection, intravenous injection, transdermal administration, and inhalation.

[0371] According to any of the foregoing aspects of the pharmaceutical formulation, the carrier or excipient is selected from one or more of buffer solutions, preservatives, isotonic agents, solubilizers, stabilizers, surfactants, and chelating agents.

[0372] According to any of the foregoing pharmaceutical formulations, the buffer solution is selected from one or more of phosphate buffer, hydrogen phosphate buffer, dihydrogen phosphate buffer, acetate buffer, carbonate buffer, bicarbonate buffer, borate buffer, maleate buffer, fumarate buffer, lactate buffer, citrate buffer, sodium barbital buffer, and tris(hydroxymethyl)aminomethane (Tris) buffer.

[0373] According to any of the foregoing pharmaceutical formulations, in some embodiments, the buffer solution is one or more of disodium hydrogen phosphate, dipotassium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, and sodium acetate solution.

[0374] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, the buffer solution is a disodium hydrogen phosphate solution.

[0375] According to any of the foregoing pharmaceutical formulations, the preservative is selected from one or more of phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, phenyl nitrite mercury, phenoxyethanol, formaldehyde, chlorobutanol, magnesium chloride (such as magnesium chloride hexahydrate), alkyl p-hydroxybenzoates (methyl ester, ethyl ester, propyl ester, butyl ester, etc.), benzalkonium chloride, benzyl chloride, sodium methyl esterpyranone, and thimerosal.

[0376] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, the preservative is phenol.

[0377] According to any of the foregoing aspects of the pharmaceutical formulation, the isotonic agent is a commonly used osmotic pressure regulator in the art, such as a carbohydrate, polyol, or salt. The isotonic agent is selected from one or more of glycerol, propylene glycol, monosaccharides, disaccharides, polysaccharides, aldehydes, and sodium chloride; preferably, the monosaccharide is selected from fructose, maltose, galactose, glucose, D-mannose, sorbitol, etc.; preferably, the disaccharide is selected from lactose, sucrose, trehalose, cellobiose, etc.; preferably, the polysaccharide is selected from raffinose, melitriose, maltodextrin, dextran, starch, etc.; preferably, the aldehyde is selected from mannitol, xylitol, maltitol, lactitol, xylitol, sorbitol (glucitol), etc. The osmotic pressure regulator can also be selected from synthetic substances, such as polyethylene glycol or polypropylene glycol, for example, PEG200, PEG400, PEG1000, PEG4000, etc.

[0378] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, the isotonic agent is glycerin.

[0379] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, the isotonic agent is propylene glycol.

[0380] In some embodiments of the pharmaceutical formulation according to any of the foregoing aspects, the excipient is a sugar excipient; the sugar excipient is selected from one or more of monosaccharides, disaccharides, polysaccharides, and aldehydes and alcohols.

[0381] According to any of the foregoing pharmaceutical formulations, in some embodiments, the surfactant is selected from polysorbates (e.g., Tween 20® (polyoxyethylene (20) sorbitan monolaurate), Tween 40® (polyoxyethylene (20) sorbitan monopalmitate), Tween 80® (polyoxyethylene (20) sorbitan monooleate)), poloxamer 184 or 188, prolonic acid F68 (polyoxyethylene-polyoxypropylene block copolymer), and PEG (polyethylene glycol) or surfactants such as polysorbate 20 or 80, or poloxamer 184 or 188, prolonic acid, and other block copolymers, one or more. Typically, the concentration of Tween 20® or Tween 80® is between 0.001% and 0.05%. Other preferred ranges for Tween 20® or Tween 80® are between 0.005% and 0.05%, between 0.0075% and 0.05%, and between 0.01% and 0.05%.

[0382] In some embodiments of the pharmaceutical formulation of any of the foregoing aspects, the concentration of the peptide compound is at least 0.1 mg / ml, preferably 0.5-20 mg / ml, 0.5-10 mg / ml, 1-8 mg / ml, 1-6 mg / ml, 1-5 mg / ml, 1.5-4 mg / ml; more preferably 1.5 mg / ml.

[0383] According to any of the foregoing aspects of the pharmaceutical formulation, in some embodiments, the concentration of disodium hydrogen phosphate is at least 0.1 mg / ml, preferably 0.1-10 mg / ml, 0.3-10 mg / ml, 0.5-10 mg / ml, 0.5-5 mg / ml, 0.5-3 mg / ml, or 0.7-2 mg / ml. The disodium hydrogen phosphate used in the formulation of the present invention can be anhydrous disodium hydrogen phosphate or disodium hydrogen phosphate hydrate, such as disodium hydrogen phosphate dihydrate, disodium hydrogen phosphate heptahydrate, disodium hydrogen phosphate dodecahydrate, etc. Disodium hydrogen phosphate dodecahydrate is preferred as a pharmaceutical excipient.

[0384] According to any of the foregoing pharmaceutical formulations, in some embodiments, the concentration of glycerol is at least 12 mg / ml, preferably 15-25 mg / ml, 17-22 mg / ml, or 17-20 mg / ml.

[0385] According to any of the foregoing pharmaceutical formulations, in some embodiments, the concentration of propylene glycol is at least 8 mg / ml, preferably 10-20 mg / ml, 10-18 mg / ml, 10-16 mg / ml, 12-16 mg / ml, or 13-15 mg / ml.

[0386] According to any of the foregoing aspects of the pharmaceutical formulation, in some embodiments, the pH of the pharmaceutical formulation is between 3 and 9, preferably 4-8.8, 5-8.6, 6-8.5, 6.5-8.5, 7.0-8.5, 7.2-8.5, 7.2-8.0, 7.4-8.0, 7.5-8.0, 7.4-7.6, or 7.5-7.6. According to any of the foregoing aspects of the pharmaceutical formulation, in some embodiments, the preservative is phenol, preferably at a concentration not exceeding 5.5 mg / ml, more preferably not exceeding 5.0 mg / ml.

[0387] According to any of the foregoing aspects of the pharmaceutical formulation, in some embodiments, the peptide compound in the pharmaceutical formulation is compound 27, 29, 220 or 221.

[0388] In one aspect, this invention relates to methods for treating or preventing the following diseases or conditions: impaired glucose tolerance (IGT), hyperglycemia, type 1 diabetes, type 2 diabetes, obesity, metabolic syndrome, and neurodegenerative diseases, particularly for delaying or preventing disease progression in type 2 diabetes, delaying the progression from impaired glucose tolerance to type 2 diabetes; delaying the progression from type 2 diabetes to insulin-dependent diabetes; treating metabolic syndrome by regulating appetite, inducing satiety, reducing food intake, and increasing energy expenditure; treating obesity or preventing overweight; preventing weight rebound after successful weight loss; treating diseases or conditions associated with overweight or obesity; treating bulimia; treating binge eating; treating dyslipidemia, atherosclerosis, hypertension, coronary heart disease, beta-blocker poisoning; and non-alcoholic fatty liver disease (NAFLD). (liverdisease) (which can be classified as simple fatty liver (SFL), non-alcoholic steatohepatitis (NASH) and its associated cirrhosis); used to inhibit gastrointestinal motility, for use in conjunction with gastrointestinal surveys using techniques such as X-ray, CT and NMR scanning; the method comprising administering to a patient an effective amount of any of the foregoing peptide compounds or pharmaceutically acceptable salts or solvates thereof or pharmaceutical compositions thereof.

[0389] In one aspect, the present invention relates to the use of pharmaceutical preparations of any of the foregoing aspects in the preparation of medicaments for lowering blood sugar or treating diabetes.

[0390] In one aspect, the present invention relates to the use of pharmaceutical preparations of any of the foregoing aspects in the preparation of medicaments for weight loss.

[0391] In one aspect, the present invention relates to the use of pharmaceutical preparations of any of the foregoing aspects in the preparation of medicaments for lowering blood lipids, preferably lowering blood lipid components selected from the following: cholesterol, triglycerides, free fatty acids, and low-density lipoprotein cholesterol.

[0392] In one aspect, the present invention relates to a method for preparing a pharmaceutical preparation according to any of the foregoing aspects, wherein the preparation method comprises the following steps:

[0393] (1) Dissolve the buffer, isotonic agent and optional preservative and surfactant in an appropriate amount of sterile water for injection to obtain a solution;

[0394] (2) Add the peptide compound of formula (VII) or its pharmaceutically acceptable salt or solvate, and adjust to the desired pH range;

[0395] (3) Add an appropriate amount of sterile water for injection to bring the formulation solution to the target volume;

[0396] (4) Use a 0.22-micron filter membrane to sterilize the preparation solution.

[0397] Preferably, the buffer is disodium hydrogen phosphate or disodium hydrogen phosphate hydrate;

[0398] Preferably, the preservative is phenol or m-cresol; more preferably, the preservative is phenol.

[0399] Preferably, the isotonic agent is glycerol or propylene glycol; more preferably, the isotonic agent is glycerol.

[0400] The inventors have obtained a novel class of peptide compounds by performing a series of structural modifications on GLP-1 receptor agonist peptide derivatives, including selecting specific amino acids, introducing new amino acids at the C-terminus of the peptide, or substituting amino acid residues at the C-terminus of the peptide, and linking unique long-acting groups to the peptide via the thiol side chain of cysteine ​​residues or the amino side chain of lysine residues at the C-terminus of the peptide. Unexpected technical effects have been achieved. Attached Figure Description

[0401] Figure 1 Tests on the reduction of blood glucose in db / db mice by compounds 1, 2, 8, 16, 18, 19, 24 and 36 of the present invention.

[0402] Figure 2 Tests on the reduction of blood glucose in db / db mice by compounds 31, 35, 38, 44 and 49 of the present invention.

[0403] Figure 3 Intraperitoneal glucose tolerance test (IPGTT) of compounds 6, 13, 14 and 26 of the present invention.

[0404] Figure 4 Tests on the weight reduction effects of compounds 4, 11, 17, 20 and 23 of this invention.

[0405] Figure 5 The fasting blood glucose changes of compounds 4, 11, 17, 20 and 23 of this invention.

[0406] Figure 6 Compounds 4, 11, 17, 20 and 23 of this invention effectively reduce TC concentration.

[0407] Figure 7 Compounds 4, 11, 17, 20 and 23 of this invention effectively reduce TG concentration.

[0408] Figure 8 The fasting blood glucose test of compounds 21, 22, 27, 28, 29 and 30 of the present invention.

[0409] Figure 9 Tests on the effects of compounds 42, 115 and 178 of the present invention on reducing blood glucose levels in db / db mice.

[0410] Figure 10 Compounds 27 and 29 of this invention effectively reduce body weight.

[0411] Figure 11 Compounds 27 and 29 of this invention effectively reduce food intake.

[0412] Figure 12 Compounds 27 and 29 of this invention effectively reduce low-density lipoprotein cholesterol (LDL-C).

[0413] Figure 13 Compounds 35 and 40 of this invention effectively reduce adipose tissue mass. Detailed Implementation

[0414] Unless otherwise stated, the following definitions apply throughout this invention. Undefined terms may be understood according to industry conventions.

[0415] "Amino acid" refers to a molecule that contains both amino and carboxyl functional groups. In α-amino acids, the amino and carboxyl groups are attached to the same carbon atom (α carbon). The α carbon may also have 1-2 additional organic substituents. Amino acids include L and D isomers and racemic mixtures. Unless otherwise specified, the amino acid residues in the polypeptide sequence of this invention are all L isomers, i.e., L-amino acids. D-amino acids are indicated by adding a lowercase "d" before the amino acid name or abbreviation, such as dK.

[0416] The amino acid sequences of this invention contain conventional single-letter or three-letter codes for naturally occurring amino acids, as well as universally accepted three-letter codes for other amino acids, such as Tic (1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid), Aib (α-aminoisobutyric acid), or GABA (γ-aminobutyric acid). Commonly used abbreviation codes for molecular structures include:

[0417] hGlu is high in glutamic acid;

[0418] α-hGlu is an L isomer of -HNCH(CO-)CH2CH2CH2COOH;

[0419] δ-hGlu is an L isomer of -HNCH(COOH)CH2CH2CH2CO-;

[0420] α-Glu is an L-isomer of -HNCH(CO-)CH2CH2COOH;

[0421] γ-Glu or gGlu is an L isomer of -HNCH(COOH)CH2CH2CO-;

[0422] α-Asp is an L isomer of -HNCH(CO-)CH2COOH;

[0423] β-Asp is an L isomer of -HNCH(COOH)CH2CO-;

[0424] β-Ala is -HN-CH2-CH2-COOH;

[0425] PEG2 is 2-(2-(2-aminoethoxy)ethoxy)acetic acid (CAS No. 134978-97-5).

[0426] In this invention, the amino acid composition of the polypeptide can be altered without significantly affecting its biological activity. For example, a polypeptide sequence may contain one or more conserved amino acid substitutions. A conserved amino acid substitution is the replacement of one amino acid residue with another amino acid residue having a similar side chain. The literature classifies amino acid residues based on the nature of their side chains. Amino acid residues containing basic side chains include lysine, arginine, and histidine; amino acid residues containing acidic side chains and amide side chains include aspartic acid, glutamic acid, asparagine, and glutamine; amino acid residues with small aliphatic, nonpolar, or weakly polar side chains include glycine, alanine, threonine, serine, and proline; amino acid residues with large aliphatic, nonpolar side chains include leucine, isoleucine, and valine; aromatic amino acid residues include phenylalanine, tryptophan, and tyrosine; and amino acid residues with sulfur-containing side chains include cysteine ​​and methionine.

[0427] In some embodiments, the derivative comprises a substituent containing a lipophilic moiety and optionally a moiety with 1-3 negative charges, wherein one of the negatively charged moieties is distal to the lipophilic moiety. In some embodiments, the substituent is attached to the side chain of an amino acid at the C-terminus of the sequence. If the C-terminus is a lysine, it is attached to the ε-amino group of the lysine residue.

[0428] As used herein, the term "treatment" includes suppressing, slowing, stopping, or reversing existing symptoms or the progression or severity of a patient's condition. Therefore, treatment encompasses prevention, treatment, and / or cure. Prevention refers to preventing underlying disease and / or preventing the worsening of symptoms or the development of disease.

[0429] As used in this article, “therapeutic effect” refers to the effect resulting from treatment of an individual, which alters, usually improves or enhances the symptoms of a disease or condition, or cures a disease or condition.

[0430] As used herein, "therapeutic effective amount" or "therapeutic effective dose" refers to the amount of a substance, compound, material, or composition containing a compound that, when applied to a subject, is at least sufficient to produce a therapeutic effect. Therefore, it is the amount necessary to prevent, cure, improve, block, or partially block the symptoms of a disease or condition.

[0431] As used herein, "preventive effective dose" or "preventive effective amount" refers to the amount of a substance, compound, material, or composition containing a compound that, when applied to a subject, would have the intended preventive effect, such as preventing or delaying the onset or recurrence of a disease or symptom, or reducing the likelihood of the onset or recurrence of a disease or symptom. A fully preventive effective dose does not necessarily occur through the administration of a single dose and can occur only after a series of doses have been administered. Therefore, a preventive effective dose can be administered in one or more applications.

[0432] As used in this article, the term "patient" refers to mammals, such as humans.

[0433] Holst (Holst, JJPhysiol.Rev.2007, 87, 1409) and Meier (Meier, JJNat.Rev.Endocrinol.2012, 8, 728) described GLP-1 receptor agonists such as GLP-1, liraglutide, and lizard exotropic peptide-4.

[0434] Some compounds of the present invention are generally effective over a wide dose range. For example, a once-weekly dose can range from about 0.05 to about 30 mg per person per week. Some compounds of the present invention can be administered daily. Additionally, some compounds of the present invention can be administered once weekly.

[0435] It should be understood that the therapeutic agent according to the described embodiment will be administered together with pharmaceutically acceptable suitable carriers, excipients, and other agents incorporated into the formulation to provide improved transfer, delivery, tolerability, etc. A large number of suitable formulations can be found in all pharmacopoeias known to medicinal chemists: Remington's Pharmaceutical Sciences (15th edition, Mack Publishing Company, Easton, Pa. (1975)), particularly Chapter 87 of Blaug and Seymour. These formulations include, for example, powders, pastes, ointments, gels, waxes, oils, lipids, lipid-containing (cationic or anionic) carriers (e.g., Lipofectin™), DNA conjugates, anhydrous slurries, oil-in-water and water-in-oil emulsions, emulsion polyethylene glycol (polyethylene glycol of various molecular weights), semi-solid gels, and semi-solid mixtures containing polyethylene glycol. Any of the foregoing mixtures may be used for the treatment or therapy according to the invention, provided that the active ingredient in the formulation is not inactivated by the formulation and the formulation is physiologically compatible and tolerable to the route of administration.

[0436] As used herein, the term "pharmaceutically acceptable carrier" is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents, and absorption delayers compatible with drug administration. Suitable carriers are described in the latest edition of Remington's Pharmaceutical Sciences, the standard bibliography in the art, which is incorporated herein by reference. Preferred examples of such carriers or diluents include, but are not limited to, water, saline, Ringer's solution, glucose solution, and 5% human serum albumin. Liposomes and non-aqueous carriers, such as immobilized oils, may also be used. The use of such media and reagents for pharmaceutically active substances is well known in the art.

[0437] Preparations intended for clinical in vivo administration must be sterile. This can be easily achieved through filtration using sterile membrane filters.

[0438] The compounds of this invention can react with any of a variety of inorganic or organic acids to form pharmaceutically acceptable acid addition salts. Pharmaceutically acceptable salts and common methods for preparing them are well known in the art. See, for example, Handbook of Pharmaceutical Salts: Properties, Selection and Use, Second Revised Edition (Wiley-VCH, 2011); SMBerge et al., “Pharmaceutical Salts”, Journal of Pharmaceutical Sciences, Vol. 66, No. 1, January 1977. Commonly used pharmaceutically acceptable salts include trifluoroacetates, acetates, citrates, hydrochlorides, phosphates, etc.

[0439] The compounds of the present invention can react with one or more inorganic or organic bases to form pharmaceutically acceptable salts. Commonly used pharmaceutically acceptable salts include sodium salts, potassium salts, ammonium salts, etc.

[0440] The pharmaceutical composition of the embodiments described herein is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, such as intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (i.e., local), transmucosal, and rectal administration. Solutions or suspensions for parenteral, intradermal, or subcutaneous administration may include the following components: sterile diluents for injection, such as water, saline solutions, fixative oils, polyethylene glycols, glycerol, propylene glycol, or other synthetic solvents; antibacterial agents, such as benzyl alcohol, methylparaben, phenol, or m-cresol; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid (EDTA); buffers, such as acetates, citrates, or phosphates; and osmotic pressure adjusting agents, such as sodium chloride or dextran. pH may be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. Parenteral formulations may be packaged in ampoules, vials, disposable syringes, glass or plastic multi-dose vials, or injection pens. There are two main types of injection pens: one is a disposable pre-filled pen containing medication, which does not require replacement of the medication cartridge and can be discarded after use; the other is a more commonly used durable injection pen, which consists of an injection pen and a medication cartridge, and can be used again by replacing the cartridge after use.

[0441] Suitable pharmaceutical compositions for injection include sterile aqueous solutions (in this case, water-soluble) or dispersions, as well as sterile powders for immediate preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, antibacterial water, Cremophor EL™ (BASF, Parsippany, NJ), or phosphate-buffered saline (PBS). In all cases, the composition must be sterile and its flowability should be sufficient for easy injection. It must be stable under manufacturing and storage conditions and must be resistant to contamination by microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof. Suitable flowability can be maintained, for example, by using a coating such as lecithin to maintain the desired particle size in the case of dispersions, and by using surfactants. Prevention of microbial action can be achieved by various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, m-cresol, ascorbic acid, thimerosal, etc. In many cases, it is preferable to include isotonic agents in the composition, such as sugars, polyols (e.g., mannitol, sorbitol), and sodium chloride. Prolonged absorption of the injectable composition can be achieved by including absorption-retarding agents such as aluminum monostearate and gelatin in the composition. If desired, a sterile injectable solution can be prepared by incorporating the compound of the invention in the desired amount into a suitable solvent having one or a combination of the ingredients listed above (as needed), followed by filtration sterilization. Generally, dispersions are prepared by incorporating the compound of the invention into a sterile carrier containing a dispersion medium and the other desired ingredients listed above. For sterile powders used to prepare sterile injectable solutions, the preparation method is to obtain a powder containing the active ingredient and any other desired ingredients derived from a sterile filtered solution of these ingredients.

[0442] For inhalation administration, the compound is delivered in the form of an aerosol spray from a pressurized container or dispenser or nebulizer containing a suitable propellant such as carbon dioxide.

[0443] Systemic administration can also be achieved via mucosal or transdermal routes. For mucosal or transdermal administration, a penetrant suitable for the permeability barrier is used in the formulation. Such penetrants are generally known in the art and include detergents, bile salts, and fusidic acid derivatives, such as those used for mucosal administration. Mucosal administration can be achieved by using nasal sprays or suppositories. For transdermal administration, one or more of the compounds of the present invention can be formulated into ointments, ointments, gels, or creams as commonly known in the art.

[0444] The compound can also be prepared in the form of suppositories (e.g., having a conventional suppository base, such as cocoa butter or other glycerides) or retention enemas for rectal delivery.

[0445] In one embodiment, the compounds of the present invention can be prepared using a carrier that prevents their rapid elimination from the body, such as a sustained-release / controlled-release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene-vinyl acetate, polyanhydride, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. Methods for preparing such formulations will be apparent to those skilled in the art.

[0446] For example, these active ingredients can be encapsulated in microcapsules, such as those prepared by coagulation technology or interfacial polymerization, for example, hydroxymethyl cellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or macroemulsions.

[0447] Sustained-release formulations can be prepared. Examples of suitable sustained-release formulations include semi-permeable matrices of solid hydrophobic polymers containing the compounds of the present invention, which are in the form of molded articles such as membranes or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (e.g., poly(2-hydroxyethyl-methylpropionate), or poly(vinyl alcohol)), polylactides (US Patent No. 3,773,919), copolymers of L-glutamic acid and γ-ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic-glycolic acid copolymers such as LUPRON DEPOT™ (injectable microspheres composed of lactic-glycolic acid copolymer and leuprolide acetate), and poly-D-(-)-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic-glycolic acid can release molecules for more than 100 days, some hydrogels release proteins for shorter periods. Polylactic acid (PLA) and polylactic-glycolic acid copolymer (PLGA) have been hot topics in recent years. In addition, there are albumin microspheres, chitosan microspheres, gelatin microspheres, etc.

[0448] Liposome suspensions can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example as described in U.S. Patent No. 4,522,811.

[0449] Particularly advantageous is the formulation of parenteral compositions in unit dosage form for ease of administration and dosage consistency. As used herein, unit dosage form refers to physically separable units suitable as unit doses for use in the subject to be treated; each unit contains a predetermined amount of one or more of the compounds of the invention calculated to bind with the desired drug carrier to produce the desired therapeutic effect. The specifications of the unit dosage form of the embodiments are indicated by and directly depend on: the unique characteristics of the compounds of the invention and the specific therapeutic effect to be achieved, and the inherent limitations in the field of formulation of such compounds of the invention for the individual to be treated.

[0450] The pharmaceutical composition may be placed in a container, package, or dispenser together with the instructions for use.

[0451] This invention provides a method for treating type 2 diabetes in patients, comprising administering an effective amount of the compound of the invention or a pharmaceutically acceptable salt thereof to a patient requiring such treatment. This invention also provides a method for treating type 2 diabetes in patients, comprising administering an effective amount of the compound of the invention or a pharmaceutically acceptable salt thereof to a patient requiring such treatment, wherein the administration is subcutaneous. This invention further provides a method for treating type 2 diabetes in patients, comprising administering an effective amount of the compound of the invention or a pharmaceutically acceptable salt thereof to a patient requiring such treatment, and simultaneously, separately, or sequentially administering effective amounts of one or more other active ingredients. In one embodiment, the other one or more active ingredients are currently available oral glucose-lowering drugs derived from a class of drugs considered as a standard of care prior to administration (as determined by industry guidelines such as those of the American Diabetes Association).

[0452] This invention also provides methods for treating or preventing the following diseases or conditions: impaired glucose tolerance (IGT), hyperglycemia, type 1 diabetes, type 2 diabetes, obesity, metabolic syndrome, and neurodegenerative diseases, particularly for delaying or preventing disease progression in type 2 diabetes, delaying the progression from impaired glucose tolerance to type 2 diabetes; delaying the progression from type 2 diabetes to insulin-dependent diabetes; treating metabolic syndrome by regulating appetite, inducing satiety, reducing food intake, increasing energy expenditure, treating obesity or preventing overweight; preventing weight rebound after successful weight loss; treating diseases or conditions associated with overweight or obesity; treating bulimia; treating binge eating; treating dyslipidemia, atherosclerosis, hypertension, coronary heart disease, and beta-blocker poisoning; non-alcoholic fatty liver disease (NAFLD) (which can be divided into simple fatty liver (SFL), non-alcoholic steatohepatitis (NASH), and related cirrhosis); and for inhibiting gastrointestinal motility, for use in conjunction with gastrointestinal surveys using techniques such as X-ray, CT, and NMR scanning. The method includes administering an effective amount of the compound of the present invention or a pharmaceutically acceptable salt thereof or a solvate thereof to a patient in need of such treatment, and simultaneously, separately, or sequentially administering effective amounts of one or more other active ingredients.

[0453] Further preferred medicinal uses include the treatment or prevention of degenerative diseases, particularly neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, ataxia (e.g., spinocerebellar ataxia), Kennedy's disease, myotonic dystrophy, Lewy body dementia, multiple systemic atrophy, amyotrophic lateral sclerosis (ALS), primary ALS, spinal muscular atrophy, prion-related diseases (e.g., Creutzfeldt-Jacob disease), multiple sclerosis, capillary aplasia, Batten disease, corticobasal degeneration, subacute combined degeneration of the spinal cord, tabes dorsalis, and Tay-Sachs disease. Diseases, toxic encephalopathy, infantile Refsum disease, neuroacanthosis, Niemann-Pick disease, Lyme disease, Machado-Joseph disease, Sandhoff disease, Shy-Drager syndrome, wobbly hedgehog syndrome, primary disease (proteopathy), cerebral β-amyloid angiopathy, retinal ganglion cell degeneration in glaucoma, synucleinopathies, tauopathies, frontotemporal lobe degeneration (FTLD), dementia, Cadasil syndrome, hereditary cerebral hemorrhage with amyloidosis, Alexander disease, seipinopathies, familial amyloid neuropathy, senile systemic amyloidosis. Amyloidosis, serpinopathies, AL (light chain) amyloidosis (primary systemic amyloidosis), AH (heavy chain) amyloidosis, AA (secondary) amyloidosis, and intra-aortic amyloidosis (aortic medial)Amyloidosis, ApoAI amyloidosis, ApoAII amyloidosis, ApoAIV amyloidosis, Finnish familial amyloidosis (FAF), lysozyme amyloidosis, fibrinogen amyloidosis, dialysis amyloidosis, inclusion body myositis / myopathy, cataract, retinitis pigmentosa with rhodopsin mutation, medullary thyroid carcinoma, atrial amyloidosis, pituitary prolactinoma, hereditary lattice corneal dystrophy, cutaneous lichenoid amyloidosis, Mallory bodies, corneal lactoferrin amyloidosis, pulmonary alveolar proteinosis, pindborg tumor amyloidosis, cystic fibrosis, sickle cell disease, or critical illness myopathy (CIM). Further medical applications include treating bone-related conditions such as osteoporosis or osteoarthritis, where increased bone formation and reduced bone resorption may be beneficial.

[0454] Abbreviations

[0455] Protective base:

[0456] Aloc or AOC, allyloxycarbonyl: allyloxycarbonyl; Bom, benzyloxymethyl: benzyloxymethyl; 2-Br-Z, 2-bromobenzyloxycarbonyl: 2-bromobenzyloxycarbonyl; tBu, t-butyl: tert-butyl; Bz, benzoyl: benzoyl; Bzl, benzyl: benzyl; Boc, tert-butoxycarbonyl: tert-butoxycarbonyl; CHO, formyl: formyl; cHx, cyclohexyl: cyclohexyl; Cbz or Z, benzyloxycarbonyl: benzyloxycarbonyl; 2-Cl-Z, 2-chlorobenzyloxycarbonyl: 2-chlorobenzyloxycarbonyl; Fm, 9-fluorenylmethyl: 9-fluorenylmethyl; Fmoc, 9-fluorenylmethoxycarbonyl: 9-fluorenylmethoxycarbonyl; Mtt, 4-methyltrityl: 4-methyltriphenylmethyl; Pmc, (2,2,5,7,8-pentamethylchroman-6-sulphonyl: 2,2,5,7,8-pentamethyl-6-hydroxychromanyl; Tos, 4-toluenesulphonyl: p-toluenesulfonyl; Trt, triphenylmethyl: triphenylmethyl; Xan,xanthyl: xanthyl, xanthyl.)

[0457] Reagents and solvents:

[0458] ACN, acetonitrile: acetonitrile; BOP, benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate: benzotriazol-1-tris(trimethylamino)-hexafluorophosphate (Carter's condensing agent); DCC, N,N'-Dicyclohexylcarbodiimide: dicyclohexylcarbodiimide; DCM: dichloromethane; DEPBT, 3-(Diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one: 3-(diethoxy-o-acyloxy)-1,2,3-benzotriazine-4-one; DIC, N,N'-Diisopropylcarbodiimide: N,N'-diisopropylcarbodiimide; DIPEA (or DIEA), diisopropylethylamine: diisopropylethylamine; DMF: N,N-dimethylformamide; DMSO: dimethyl sulfoxide; EDC or EDCI, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide: 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride; EtOAc: ethyl acetate; HATU, 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate: 1-[di(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate; HBTU, O-(1H-benzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate: benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate; HOAT, 1-Hydroxy-7-azabenzotriazole: 1-hydroxy-7-azabenzotriazole; HOBT, 1-hydroxybenzotriazole; Cl-HOBT; NMM, N-Methylmorpholine; NMP N-methylpyrrolidinone: N-methylpyrrolidone; Su,succinimide: succinimide; TEA, triethylamine: triethylamine; TFA, trifluoroacetic acid; TIS, triisopropylsilane: triisopropylsilane.

[0459] Polypeptide chemical synthesis methods

[0460] Solid-phase chemical synthesis of peptides is a well-developed methodology, as can be found in references such as R.C. Sheppard, Solid Phase Peptide Synthesis. A Practical Approach, Oxford-IRL Press, New York, 1989.

[0461] Linear peptides are synthesized using either the Boc solid-phase peptide synthesis method or the Fmoc solid-phase peptide synthesis method. When using Fmoc chemistry to synthesize peptides with a C-terminal carboxyl group, Wang resin is typically chosen; for peptides with a C-terminal amide group, Rink amide resin is typically chosen (in this article, Rink amide resin includes Rink Amide-AM resin, Rink Amide-MBHA resin, and similar resins). When using Boc chemistry to synthesize peptides with a C-terminal carboxyl group, Pam resin is typically chosen; for peptides with a C-terminal amide group, MBHA resin is typically chosen. Commonly used condensing agents and activators are DIC and HOBT; other optional peptide bond condensing agents include BOP, HBTU, and DEPBT. A 5-fold excess of amino acids is used. The condensation time is 1 hour. Peptides can be synthesized manually or using a peptide solid-phase synthesizer.

[0462] The Fmoc protecting group was removed using 20% ​​piperidine / DMF. The Boc protecting group was removed using TFA. The peptide bond condensation reaction was monitored using ninhydrin (2,2-Dihydroxyindane-1,3-dione) reagent.

[0463] Solid-phase synthesis can use resins pre-loaded with C-terminal amino acids or resins without amino acid loading.

[0464] The method for loading the first amino acid onto Rink Amide resin can be found in industry practice. A common method is briefly described below: Weigh an appropriate amount of resin and remove the Fmoc protecting group in a solid-phase synthesis tube using 20% ​​piperidine / DMF (15 mL / g resin, 30 min x 2). Wash the resin with DMF. Weigh an amount of Fmoc amino acid equivalent to 5 times the resin amino group, HATU, HOAT, and NMM equivalent to 10 times the resin amino group. Add DMF, mix well, and transfer to a solid-phase synthesis tube. React overnight, then wash the resin with DMF. Add a 1:1 acetic anhydride / pyridine (v / v) mixture to the solid-phase synthesis tube, drain after 30 minutes, and wash the resin with DMF. The first amino acid is now loaded.

[0465] The commonly used amino acids and protecting groups in Fmoc solid-phase peptide synthesis are as follows:

[0466] Fmoc-Cys(Trt)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-His(Trt)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Asn(Trt)-O H, Fmoc-Gln(Trt)-OH, Fmoc-Arg(Pmc)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Tyr(tBu)-OH

[0467] The synthesis utilizes appropriately protected structural units, such as the aforementioned standard amino acids, Fmoc-8-amino-3,6-dioxanoic acid (CAS No. 166108-71-0), and Fmoc-Glu-OtBu (CAS No. 84793-07-7). The introduction of the fatty acid moiety can be achieved using structural units, such as, but not limited to, eicosanoic acid monotert-butyl ester. After each coupling step, the unreacted peptide intermediate can be capped with acetic anhydride (10 equivalents) and excess chlorhexidine (20 equivalents).

[0468] After solid-phase Fmoc chemical synthesis of peptides, the commonly used cleavage reagent is TFA. Dry resin is placed in a shake flask, and an appropriate amount of cleavage buffer containing 90:4:2:2:2 (v / v) trifluoroacetic acid:triisopropylsilane:1,2-ethylenedithiol:water:benzyl sulfide (10-25 mL / g resin) is added. The flask is capped, and the mixture is subjected to intermittent rotary shaking at room temperature. After 2 hours, the resin is filtered, and washed 2-3 times with fresh TFA. The filtrates are combined, and 8-10 times their volume of ice-cold diethyl ether is added dropwise. Finally, the precipitated crude peptide is collected by centrifugation.

[0469] When using the Boc solid-phase polypeptide synthesis method, commonly used amino acids and protecting groups are as follows: Boc-Cys(4-MeBzl)-OH, Boc-Asp(OcHx)-OH, Boc-Glu(OcHx)-OH, Boc-His(Bom)-OH, Boc-Lys(2-Cl-Z)-OH, Boc-Asn(Xan)-OH, Boc-Arg(Tos)-OH, Boc-Ser(Bzl)-OH, Boc-Thr(Bzl)-OH, Boc-Trp(CHO)-OH, and Boc-Tyr(2-Br-Z)-OH.

[0470] If the side-chain amino group of lysine is used for the synthesis of lactams or acylation reactions, the side-chain amino group of lysine can be protected with an allyloxycarbonyl (aloc) or Fmoc. If the side-chain carboxyl group of aspartic acid or glutamic acid is used for the synthesis of lactams or acylation reactions, the carboxyl group should be converted to an allyl ester or 9-fluorenylmethyl for protection, such as Boc-Glu(OAllyl)-OH or Boc-Glu(Ofm)-OH.

[0471] After solid-phase Boc chemical synthesis of peptides, HF cleavage is typically performed on PAM and MBHA resins. 5 mL of HF is added for every 0.1 mmol of resin, along with reagents such as p-cresol, p-mercaptophenol, or anisole. The mixture is stirred in an ice bath for 1 hour. After vacuum drying of the HF, the peptides are precipitated with ice-cold diethyl ether. The precipitate is collected by centrifugation, purified by HPLC, and then freeze-dried to obtain the final product.

[0472] purification

[0473] The crude peptide was dissolved in a suitable mixture of water and acetonitrile (e.g., water / acetonitrile (3:1)) and purified by reversed-phase preparative HPLC (e.g., AKTA purifier, Shimadzu LC-20AR, etc.). Different packing materials and column sizes were selected based on the amount of crude peptide loaded and its polarity, such as C8 or C18 semi-preparative or preparative columns. Buffer A was 0.1% TFA aqueous solution, and buffer B was 0.1% TFA in acetonitrile. Elution was performed using a gradient uplift of buffer B, and the fractions were examined by analytical HPLC. A ZORBAX 300 SB-C18 (4.6 x 250 mm, 5 µM) column was used, with buffer A being 0.1% TFA aqueous solution and buffer B being 0.1% TFA in acetonitrile. The flow rate was 1 ml / min, and detection was performed at 210 nm. The fractions containing the purified target peptide were mixed and freeze-dried to obtain the peptide product, which was then aliquoted into glass vials for storage.

[0474] Preparation method

[0475] The compounds of this invention are linear peptides. Each amino acid can be sequentially coupled from the C-terminus to the N-terminus of the polypeptide sequence to obtain the polypeptide backbone. The process is as follows: First, an amino acid with one amino group protected by a blocking group is covalently linked to a solid support. The amino protecting group of the first amino acid is then removed, thus attaching the first amino acid to the solid support. Next, the carboxyl group of the second amino acid with its amino group blocked is activated and reacts with the amino group of the first amino acid already attached to the solid support to form a peptide bond, thereby generating a dipeptide with a protecting group on the solid support. This peptide bond formation reaction is repeated to elongate the peptide chain from the C-terminus to the N-terminus until the desired peptide chain is formed. Finally, the protecting group is removed, and the covalent bond between the peptide chain and the solid support is hydrolyzed to obtain the synthesized peptide.

[0476] Some compounds of this invention are synthesized by conjugating long-acting groups or modifying groups to the side chains of amino acids containing amino or thiol groups. Compound 27 is used as an example to illustrate the synthetic route and method.

[0477] The synthesis of compound 27 includes the following steps:

[0478] Step A: Couple Lys(PG) and resin to obtain Lys(PG)-resin, where PG is the protecting group of the amino group of the lysine side chain;

[0479] Step B: Lys(PG)-resin is first stepwise coupled with an amino acid or amino acid derivative to obtain a first peptide resin with the sequence shown in the main peptide chain of compound 27.

[0480] Step C: Remove the side chain protecting group PG of Lys from the first peptide resin, as shown in the main peptide chain of compound 27, and then proceed with a second stepwise coupling of 2-(2-(2-aminoethoxy)ethoxy)acetic acid, 2-(2-(2-aminoethoxy)ethoxy)acetic acid, γ-Glu and octadecanoic acid to obtain the second peptide resin;

[0481] Step D: The second peptide resin was cleaved and purified to obtain compound 27.

[0482] Preferably, the resin in step A is Rink Amide resin, or similar resins such as Rink Amide-AM resin and Rink Amide-MBHA resin. Preferably, the activator used for coupling in step A is selected from DIC and HOBt, or HATU and HOAT, or BOP, PyBOP, PyAOP, HBTU, TBTU, DEPBT, etc. Preferably, the side-chain protecting group of Lys is aloc, Dde, or ivDde.

[0483] In step B, during the first stepwise coupling, based on the sequence of compound 27, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Ala-OH, Fmoc-Gly-OH, Fmoc-Ser(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Gly-OH, Fmoc-Gly-OH, Fmoc-Ala-OH, Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Trp(Boc)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Ile–OH, Fmoc-Phe-OH, Fmoc-Leu-OH, and Fmoc-Arg(Pbf)-OH are sequentially coupled. Fmoc-Val-OH, Fmoc-Ala-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Leu-OH, Fmoc-Ala-OH, Fmoc-Ile–OH, Fmoc-Ser(tBu)-OH, Fmoc-Leu-O H, Fmoc-Asp(OtBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Phe-OH, Fmoc-Thr(tBu)-OH, Fmoc-Gly-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Aib-OH and Boc-Tyr(tBu)-OH.

[0484] The coupling agents used in the first stepwise coupling in step B and the second stepwise coupling in step C include condensing agents and reaction solvents. The condensing agent can be a mixture of DIC and HOBt, a mixture of PyBOP, HOBt and DIEA, or a mixture of HATU, HOAt and DIEA, or a mixture of DEPBT and DIEA, or a mixture of HBTU and DIEA. The reaction solvent is one or a mixture of two or more of DMF, DCM, NMP or DMSO.

[0485] When PG is alooc, the reagent used for removal in step C is 1-20 equivalents of morpholine (or 1-20 equivalents of benzylsilane instead of morpholine) and 0.05-3 equivalents of Pd(PPh3)4. Preferably, the alooc protecting group is removed using 5-10 equivalents of morpholine (or 5-10 equivalents of benzylsilane instead of morpholine) and 0.1-0.3 equivalents of Pd(PPh3)4. The protecting group removal reaction can be carried out twice, each time for 10-30 minutes, with CH2Cl2 preferably used as the solvent.

[0486] Another method for removing the Aloc protecting group is to stir for 2 hours at room temperature under argon atmosphere with a catalytic amount of tetra(triphenylphosphine)palladium(0) and a 37:2:1 ratio of DCM, glacial acetic acid, and NMM (15 mL / g resin). After the reaction, each gram of resin is washed with 0.5% DIPEA / DMF (10 mL), 0.5% sodium diethyldithiocarbamate / DMF (3 x 10 mL), and 1:1 DCM:DMF (5 x 10 mL).

[0487] When PG is Dde or ivDde, the reagent used for deprotection in step C is a hydrazine hydrate / DMF mixture. Prepare a 2% (w / v) hydrazine hydrate DMF solution (25 mL / g resin), add it to the resin, dry it after 5 minutes, and wash the resin with DMF. Repeat the process of deprotection with 2% hydrazine hydrate / DMF and washing with DMF three times.

[0488] In step C, during the second stepwise coupling, Fmoc-8-amino-3,6-dioxanoic acid (CAS No. 166108-71-0), Fmoc-Glu-OtBu (CAS No. 84793-07-7), and HOOC-(CH2)16-COOtBu monotert-butyl octadecanoate are sequentially coupled according to the sequence of compound 27.

[0489] The reagents used for cleavage in step D include one, two, or more compounds selected from TFA and PhSMe, PhOMe, EDT, H2O, TIS, and PhOH. For example, the reagent used for cleavage is a mixture of TFA, anisole, dimethyl sulfide, and EDT, with a volume ratio of 90:5:3:2. Preferably, the reagent used for cleavage is a mixture of TFA, H2O, and TIS, with a volume ratio of 95:2.5:2.5.

[0490] This invention also includes novel intermediates and methods for synthesizing the compounds of the invention or pharmaceutically acceptable salts thereof. The intermediates and compounds of the invention can be prepared by a variety of methods known in the art. In particular, the following examples illustrate methods using chemical synthesis. The specific synthetic steps of each described route can be combined in different ways to prepare the compounds of the invention or salts thereof. Reagents and raw materials are readily available to those skilled in the art.

[0491] Example

[0492] The invention is further illustrated with reference to the embodiments. These embodiments are not intended to limit the scope of the claimed invention. Those skilled in the art can draw upon the content of this invention, appropriately modify process parameters, and make modifications or appropriate changes and combinations to the methods and applications described in this invention to implement and apply the technology of this invention. All similar substitutions and modifications will be apparent to those skilled in the art, and they are all considered to be included in this invention.

[0493] Amino acid raw materials and condensation reagents were purchased from Jier Biochemical (Shanghai) Co., Ltd. Rink Amide resin was purchased from Shangyu Puer Company and Tianjin Nankai Hecheng Technology Co., Ltd.

[0494] Example 1: Synthesis of polypeptide 2

[0495] Pre-loaded low-to-medium loading Fmoc-Ser(tBu)-Rink Amide resin (e.g., 0.4 mmol / g) was selected. The Fmoc protecting groups were removed with 30% piperidine / DMF (2 x 10 min), and the resin was washed three times with DMF. The Fmoc solid-phase peptide synthesis method was used. Amino acids (10 equivalents relative to the resin amino acid loading) were dissolved in DMF to prepare a 0.3 M solution. HOBT and DIC (10 equivalents relative to the resin amino acid loading) were added to the solution, and the mixture was shaken to mix thoroughly before adding it to the resin. Coupling was performed at room temperature for 60 min. The solution in the solid-phase reaction tube was drained, and the resin was washed three times with DMF. This completes the coupling of one synthetic unit. Each synthetic unit of the peptide from the C-terminus to the N-terminus was coupled using the same method, with amino acid residues protected using the commonly used Fmoc solid-phase synthesis protecting groups listed in the "Solid-Phase Chemical Synthesis Methods for Peptides" section, or other protecting groups suitable for this sequence.

[0496] After peptide synthesis, the resin was washed with DCM, and a cleavage solution containing 90:4:2:2:2 (v / v) trifluoroacetic acid:triisopropylsilane:1,2-ethylenedithiol:water:benzyl sulfide (10-25 mL / g resin) was added. The mixture was shaken at room temperature for 2 hours and precipitated with ice-cold diethyl ether. The crude peptide was dissolved in 0.1% TFA and 30% acetonitrile aqueous solution and eluted using a preparative RP-HPLC on a C8, 5µM reversed-phase column. Buffer A was 0.1% TFA aqueous solution, and buffer B was 0.1% TFA in acetonitrile. Buffer B was used for elution via a gradient, and the correct fractions were combined and freeze-dried at low temperature to obtain a white solid. The structure of the peptide was determined by mass spectrometry and amino acid sequencing. Compounds with a serine residue at the C-terminus in this invention can be synthesized according to the method in Example 1.

[0497] Example 2 Synthesis of Peptide 7

[0498] Pre-loaded Fmoc-Gly-Rink Amide resin was selected. The remaining synthesis methods and steps were similar to those in Example 1. Compounds with glycine at the C-terminus in this invention can be synthesized according to the method in Example 2.

[0499] Example 3 Synthesis of Compound 23

[0500] Pre-loaded low-load Fmoc-Lys(Aloc)-Rink Amide resin (e.g., 0.29 mmol / g) was selected. Each synthetic unit of the peptide from the C-terminus to the N-terminus was coupled using the method described in Example 1, with the amino acid residues other than the C-terminal lysine using the protecting groups commonly used in Fmoc solid-phase synthesis listed in the "Solid-Phase Chemical Synthesis of Peptides" section. After peptide synthesis, the allyloxycarbonyl group of the C-terminal lysine side chain was removed by stirring in an argon atmosphere at room temperature for 2 hours with tetrakis(triphenylphosphine)palladium(0) and a 37:2:1 ratio of DCM, glacial acetic acid, and NMM (15 mL / g resin). The tetrakis(triphenylphosphine)palladium(0) could be used in amounts up to 1 equivalent. After the reaction, each gram of resin was washed with 0.5% DIPEA / DMF (10 mL), 0.5% sodium diethyldithiocarbamate / DMF (3 x 10 mL), and a 1:1 DCM:DMF mixture (5 x 10 mL). The Aloc protecting group can also be removed using 5-10 equivalents of morpholine (or 5-10 equivalents of benzenesilane) and 0.1-0.3 equivalents of Pd(PPh3)4. The protecting group removal reaction can be carried out twice, 30 minutes each time, using CH2Cl2 as the solvent. After removing the Aloc protecting group from the side chain, the lysine side chain substituent is synthesized using the structural units Fmoc-8-amino-3,6-dioxanoic acid (CAS No. 166108-71-0), Fmoc-Glu-OtBu (CAS No. 84793-07-7), and HOOC-(CH2) octadecanoic acid monotert-butyl ester. 16 -COOtBu. The amount of each side chain substituent in each structural unit is 10 equivalents of the resin amino acid loading, and the coupling time for each step is 4 hours. The remaining synthesis methods and steps are similar to those in Example 1.

[0501] Another method involves using pre-loaded, low-load Fmoc-Lys(ivDde)-Rink Amide resin. The synthesis of the main peptide chain is the same as the method using Fmoc-Lys(Aloc)-Rink Amide resin described in the previous section. After the main peptide chain synthesis, the C-terminal lysine side chain ivDde protecting group is removed using 2% hydrazine hydrate / DMF. A 2% (w / v) hydrazine hydrate DMF solution (25 mL / g resin) is prepared, added to the resin, dried under vacuum after 5 minutes, and the resin is washed with DMF. The process of removing the protecting group with 2% hydrazine hydrate / DMF and washing with DMF is repeated three times. The synthesis method of this lysine side chain substituent is the same as the side chain linking method of Fmoc-Lys(Aloc)-Rink Amide resin described in the previous section.

[0502] Compounds in this invention with a C-terminus of lysine and a side chain amino group connected to a long-acting group, such as compounds 4, 6, 11, 13, 14, 17, 21-23, 27-30, 220 and 221, can also be synthesized using the method of Example 3, or by using the synthesis method of compound 27 in the "Preparation Method" section.

[0503] Example 4 Synthesis of polypeptide 33

[0504] Pre-loaded Fmoc-Cys(Trt)-Rink Amide resin was selected. The remaining synthesis methods and steps were similar to those in Example 1. Compounds with a C-terminus of cysteine ​​in this invention can be synthesized according to the method in Example 4.

[0505] Example 5 Synthesis of Polypeptide 95

[0506] Pre-loaded Fmoc-Pro-Rink Amide resin was selected. The remaining synthesis methods and steps were similar to those in Example 1. Compounds with a proline terminal in this invention can be synthesized according to the method in Example 5.

[0507] Example 6 Synthesis of polypeptide 110

[0508] Pre-loaded Fmoc-Ala-Rink Amide resin was selected. The remaining synthesis methods and steps were similar to those in Example 1. Compounds with an alanine terminal in this invention can be synthesized according to the method in Example 6.

[0509] Table 1 lists the calculated and measured molecular weights of some of the compounds in this invention.

[0510] Table 1

[0511]

[0512] Mass spectrometry molecular weight and amino acid sequencing results confirmed the correct peptide structure.

[0513] The peptides were dissolved in physiological saline (pH 7.4) to prepare a stock solution. The peptide concentration in the stock solution was quantified using conventional methods such as the Bradford method and ultraviolet spectrophotometry. Before animal efficacy studies, a stock solution containing the required dose of peptides was prepared and diluted with buffered physiological saline (PBS, pH 7.4) to prepare an injection solution. The appropriate injection volume per animal was 5 ml / kg body weight. This can be used to calculate the required volume of injection solution.

[0514] All data from animal experiments will be entered into an Excel document and expressed as mean ± SEM. Differences among multiple groups will be compared using GraphPad Prism 6 software using Dunnett's one-way analysis of variance (ANOVA). Differences between two groups will be compared using an unpaired T-test. A p-value less than 0.05 is considered statistically significant.

[0515] Example 7

[0516] The in vivo efficacy of the peptides of this invention can be determined in any suitable animal model known in the art, as well as in clinical trials. For example, the db / db mouse is a suitable animal model for diabetes.

[0517] db / db mice were housed in strictly controlled environments, with the temperature maintained at 20–24°C and humidity at 40–70%. Temperature and humidity were monitored in real-time using a thermometer and hygrometer, and recorded twice daily (once in the morning and once in the afternoon). Lighting in the enclosures was controlled by an electronic timer system, with lights on for 12 hours and off for 12 hours daily (starting at 7:00 AM and ending at 7:00 PM). During the experiment, animals were housed individually, with toys provided to each mouse in its cage. Animals had free access to water. Male db / db mice (6 weeks old) were acclimatized to the experimental environment for one week. Baseline blood glucose and body weight were recorded for the first three days of the experiment (days -3 to -1). Based on these three-day blood glucose and body weight data, mice were randomly assigned to groups of six. Mice were subcutaneously injected at 10:00 AM with physiological saline (5 ml / kg, control group) or peptide compounds 1, 2, 8, 16, 18, 19, 24, and 36 (30 nmol / kg). Blood samples were collected at 0 hours before administration and at 1, 2, 4, 6, 8, and 10 hours after administration. Blood glucose levels were measured using a Johnson & Johnson OneTouch blood glucose meter and matching test strips. Blood glucose curves were plotted with time on the x-axis and blood glucose values ​​at different time points on the y-axis. The area under the curve (AUC) was calculated to compare the time and efficacy of the peptide compounds in lowering blood glucose.

[0518] The results are as follows Figure 1 As shown, compounds 1, 2, 8, 16, 18, 19, 24, and 36 significantly reduced blood glucose levels in type 2 diabetic mice, showing a statistically significant difference compared to the control group. These compounds have the potential to serve as therapeutic agents for diabetes.

[0519] Example 8

[0520] The db / db mice were housed under the same conditions as in Example 7. Male db / db mice (8 weeks old) were acclimatized to the experimental environment for one week. Baseline blood glucose and body weight were recorded for the first three days of the experiment (days -3 to -1). Based on the three-day blood glucose and body weight, the mice were randomly assigned to groups of 6 mice each. Mice were subcutaneously injected at 10:00 AM with physiological saline (5 ml / kg, control group) or peptide compounds 31, 35, 38, 44, and 49 (30 nmol / kg). Blood samples were collected at 0 hours before administration and at 1, 2, 4, 6, 8, and 10 hours after administration. Blood glucose levels were measured using a Johnson & Johnson OneTouch blood glucose meter and matching test strips. A time-effect curve for the hypoglycemic effect was constructed with time on the x-axis and blood glucose values ​​at different time points on the y-axis, and the area under the curve (AUC) was calculated.

[0521] The results are as follows Figure 2 As shown, compounds 31, 35, 38, 44, and 49 significantly reduced blood glucose levels in type 2 diabetic mice, with statistically significant differences compared to the control group. These compounds have the potential to serve as therapeutic agents for diabetes.

[0522] Example 9

[0523] The db / db mice were housed under the same conditions as in Example 7. Baseline blood glucose and body weight of male db / db mice (6 weeks old) were recorded for three days prior to the experiment (days -3 to -1). Mice were randomly assigned to groups of 6 mice each based on their three-day blood glucose and body weight. At 18:00 the day before the experiment (day -1), the animals were subcutaneously injected with physiological saline (5 ml / kg, control group) or compounds 6, 13, 14, and 26 (3 nmol / kg). On the day of the experiment, the animals were fasted for 6 hours (8:00-14:00) and underwent an intraperitoneal glucose tolerance test (IPGTT). At 14:00, the animals received a single intraperitoneal injection of glucose (1.5 g / kg) aqueous solution, and the time of glucose administration was recorded as 0:00. Blood glucose levels were measured 0 minutes before glucose administration and at 15, 30, 60, 120, and 180 minutes after glucose administration. A blood glucose curve was plotted with time on the x-axis and blood glucose values ​​at different time points on the y-axis.

[0524] The results are as follows Figure 3 As shown, compounds 6, 13, 14 and 26 can significantly reduce blood glucose in type 2 diabetic mice and exhibit prolonged in vivo duration of action, showing potential as long-acting diabetes treatments.

[0525] Example 10

[0526] The in vivo efficacy of the peptides of this invention can be determined in any suitable animal model known in the art, as well as in clinical trials. Diet-induced obese (DIO) mice are animal models of obesity, insulin resistance, and hyperlipidemia.

[0527] Five-week-old male C57BL / 6 mice were housed in an animal enclosure with strictly controlled environmental conditions. The temperature was maintained at 20–24°C, and the humidity at 30–70%. Temperature and humidity were monitored in real-time using a thermometer and hygrometer, and recorded twice daily (once in the morning and once in the afternoon). Lighting in the enclosure was controlled by an electronic timer system, with lights on for 12 hours and off for 12 hours daily (starting at 6:00 AM and ending at 6:00 PM). During the experiment, animals were housed individually, and each mouse was provided with toys. Animals were fed a high-fat diet from six weeks of age (the weight ratio of nutrients was 26.2% protein, 26.3% carbohydrates, and 36.9%, providing 20%, 20%, and 60% of the total calories, respectively). Animals had free access to water. Two-five-week-old male DIO mice with an average weight of approximately 49 grams were selected for the experiment. Animals underwent one week of acclimatization by handling and subcutaneous injection. Animals were weighed and fed for three consecutive days before the experiment. Blood glucose levels were measured one day before the experiment. Animals were then grouped according to their blood glucose and weight, with six animals in each group.

[0528] During the experiment, animals in the control and liraglutide groups received subcutaneous injections of either saline or liraglutide (100 nmol / kg) once daily. The other five groups received subcutaneous injections of peptides 4, 11, 17, 20, and 23 (25 nmol / kg) on ​​days 1, 3, 5, 7, 9, 11, and 13, respectively. Animal weight and food intake were measured daily throughout the 14-day experiment. The percentage change in body weight was calculated as [(final body weight - initial body weight) / initial body weight] * 100%.

[0529] Fasting blood glucose levels in the animals were measured using a blood glucose meter one day before the experiment and on day 15. For fasting blood glucose testing, the animals were fasted from 9:00 AM to 3:00 PM, and the test was conducted at 3:00 PM.

[0530] On day 16, the animals were fasted for 6 hours (8:00-14:00), euthanized, and dissected. Blood was collected from the heart, centrifuged, and aliquoted into plasma for biochemical analysis, primarily triglycerides (TG) and total cholesterol (TC). Biochemical analysis was performed using a Hitachi 7180 fully automated biochemical analyzer. Additionally, liver samples were collected for hepatic triglyceride and cholesterol testing.

[0531] The results of the animals' weight change over 14 days are as follows: Figure 4 As shown in the figure. Compared with the control group, the weight of the treatment group decreased significantly, and the weight of the group on day 14 was statistically significantly different from that of the control group. Moreover, the weight-reducing ability of peptides 4, 11, 17, 20 and 23 was significantly better than that of the weight-loss drug liraglutide.

[0532] Changes in fasting blood glucose (comparison of fasting blood glucose before and after the test) are as follows: Figure 5 As shown, fasting blood glucose levels were significantly lower in the treatment groups compared to the control group, and the peptide groups 4, 11, 17, 20, and 23 were superior to the liraglutide group. The decrease in fasting blood glucose indicates improved glucose regulation in the treatment groups.

[0533] like Figure 6 and 7 As shown, compared with the control group, the cholesterol and triglyceride levels of the animals in the drug-treated groups decreased significantly, and the decrease in peptides 4, 11, 17, 20 and 23 was significantly greater than that in the liraglutide group.

[0534] Example 11

[0535] Male db / db mice (Jiangsu Jicui Pharmaceutical Biotechnology, 6 weeks old) were housed in a strictly controlled environment after arrival at the facility. The temperature was maintained at 20–24°C and the humidity at 40–70%. Temperature and humidity were monitored in real-time using a thermometer and hygrometer, and recorded twice daily (once in the morning and once in the afternoon). Lighting in the animal housing was controlled by an electronic timed lighting system, with lights on for 12 hours and off for 12 hours daily (7:00 AM to 7:00 PM). During the experiment, animals were housed individually, and each mouse was provided with toys. Animals had free access to water. Animals were allowed two weeks to acclimatize. For the first three days of the experiment (days -3 to -1), animals were fasted for 6 hours at 9:00 AM, and fasting blood glucose was measured at 3:00 PM. Based on three days of fasting blood glucose and body weight, mice were randomly assigned to groups of six. The experimental objective was to measure the ability of the compound to lower fasting blood glucose in mice. During the experiment, the animals were fasted from 9:00 AM to 9:00 PM. On the first day of the experiment, mice were fasted for 6 hours prior to administration (9:00 AM - 3:00 PM), and then subcutaneously injected with physiological saline (5 ml / kg) or compounds 21, 22, 27, 28, 29, and 30 (5 nmol / kg). Blood samples were collected at 0 hours before administration and at 1, 3, 6, 24, 30, and 48 hours after administration. Blood glucose levels were measured using a Johnson & Johnson OneTouch blood glucose meter and matching test strips. A time-effect curve of hypoglycemic effect was established with time as the x-axis and blood glucose values ​​at different time points as the y-axis.

[0536] The results are as follows Figure 8As shown, compounds 21, 22, 27, 28, 29, and 30 significantly reduced blood glucose levels in db / db mice, showing a statistically significant difference compared to the control group. These compounds have the potential to serve as long-acting diabetes treatments. Furthermore, three pairs of compounds, 21 and 22, 27 and 28, and 29 and 30, demonstrated that the removal of a serine residue at the adjacent position significantly enhanced the efficacy of the compounds when a long-acting group was attached to the C-terminal lysine side chain, producing unexpected effects.

[0537] Example 12

[0538] The db / db mice were housed under the same conditions as in Example 7. Male db / db mice (Jiangsu Jicui Pharmaceutical Biotechnology, 7 weeks old) were acclimatized to the experimental environment for one week. Baseline blood glucose and body weight were recorded for the first three days of the experiment (-3 days to -1 day). Based on the three-day blood glucose and body weight, the mice were randomly divided into groups of 6 mice each. Mice were subcutaneously injected with physiological saline (5 ml / kg, control group) or peptide compounds 42, 115, and 178 (35 nmol / kg) at 10:00 AM. Blood glucose was collected at 0 hours before administration and at 1.5, 4, 7, and 10 hours after administration, and measured using a Johnson & Johnson OneTouch blood glucose meter and matching test strips. Blood glucose curves were plotted with time on the x-axis and blood glucose values ​​at different time points on the y-axis. The area under the curve (AUC) was calculated to compare the time and effect of the peptide compounds in lowering blood glucose.

[0539] The results are as follows Figure 9 As shown, compounds 42, 115, and 178 significantly reduced blood glucose levels in type 2 diabetic mice. These compounds have the potential to be used as therapeutic agents for diabetes.

[0540] Example 13

[0541] The DIO obese mouse experiment was conducted by Shanghai WuXi AppTec Co., Ltd. Male C57BL / 6 mice were fed a high-fat diet (catalog number: D12492i, Research Diets, Inc.) for 25 weeks from 5 weeks of age. At 30 weeks of age, animals underwent a one-week acclimatization period, including handling and subcutaneous injection. Body weight and food intake were measured for three consecutive days prior to the experiment. Fasting blood glucose was measured one day before the experiment, and animals with excessively high blood glucose were removed. Animals were then grouped into groups of five based on random blood glucose and body weight (body weight as the primary reference indicator). During the experiment, animals were subcutaneously injected daily with saline, compound 27, and compound 29 (3 nmol / kg) for 10 days. Body weight and food intake were measured daily during the experiment. After the experiment, animals were fasted for 6 hours, euthanized, dissected, and blood was collected from the heart. The blood was centrifuged and aliquoted into plasma for biochemical analysis.

[0542] like Figure 10As shown, the weight of the animals in the treatment group decreased significantly compared with the control group. On day 11, the weight of the animals in the treatment group was significantly different from the initial weight of the animals in the same group and also significantly different from the weight of the animals in the control group (P<0.0001).

[0543] like Figure 11 As shown, compared with the control group, the food intake of the animals in the treatment group was significantly reduced. The cumulative food intake of the animals over 10 days was statistically significant compared with the control group (P<0.0001), indicating that compounds 27 and 29 have an appetite-suppressing effect.

[0544] like Figure 12 As shown, compounds 27 and 29 significantly reduced low-density lipoprotein cholesterol (LDL-C) in experimental animals, with statistically significant differences compared to the control group (P<0.0001). Since LDL-C levels are closely related to cardiovascular disease, compounds 27 and 29 have the potential to be used in the treatment of hyperlipidemia and to reduce the risk of cardiovascular disease.

[0545] Example 14

[0546] The DIO obese mouse experiment was conducted by Shanghai WuXi AppTec Co., Ltd. Animal husbandry was the same as in Example 13. Animals underwent a one-week acclimatization period at 40 weeks of age, including handling and subcutaneous injection. Body weight and food intake were measured for three consecutive days prior to the experiment. Fasting blood glucose was measured one day before the experiment, and animals with excessively high blood glucose were removed. Animals were then grouped according to their random blood glucose and body weight (body weight as the primary reference indicator), with an average body weight of 47 grams and six animals per group. During the experiment, animals were subcutaneously injected once daily with saline or 1.5 nmole of compound 35 or compound 40 per animal, for a 14-day period. Body fat percentage was measured using an EchoMRI body fat analyzer before and after the experiment.

[0547] The changes in adipose tissue and lean tissue mass in DIO obese mice before the experiment (day 0) and after the experiment (day 15) are shown in the following figures. Figure 13 As shown in the figure, compared with the control group, the amount of adipose tissue in the treatment group was significantly reduced, showing a statistically significant difference, but the amount of lean tissue remained basically unchanged, indicating that compounds 35 and 40 can achieve weight loss in a relatively ideal manner.

[0548] Example 15: Screening of salt systems for compound 27 formulations and investigation of their solubility.

[0549] 1. Disodium hydrogen phosphate system

[0550] Disodium hydrogen phosphate system: Phosphate solutions with a pH range of 6.2-8.5 were prepared, and the active pharmaceutical ingredient (API) was added to investigate its solubility. Experimental results at different pH values ​​showed that the solubility of compound 27 increased significantly with increasing pH. When pH ≤ 6.5, the solubility was below 1 mg / ml; when pH > 6.5, both solubility and dissolution rate increased significantly, reaching a concentration of 2 mg / ml or even higher. Therefore, the disodium hydrogen phosphate system with pH > 6.5 is suitable for preparing the API at the target concentration for injection.

[0551] 2. Acetate system

[0552] Acetate system: When pH > 6.5, the solubility and dissolution rate of the active pharmaceutical ingredient (API) are relatively low. A solution prepared at a concentration of 1 mg / ml takes more than 30 minutes to dissolve, making it unsuitable for scale-up production. Furthermore, drug precipitation was observed when the solution was placed under refrigeration to assess its physical stability. When pH < 6.5, dissolution is difficult.

[0553] Conclusion: A disodium hydrogen phosphate system with pH > 6.5 was selected for further research.

[0554] Example 16 Stability study of compound 27 formulation

[0555] A phosphate solution of compound 27 was prepared at a concentration of 1 mg / ml, containing 20 mg glycerol, 5.5 mg phenol, 2.86 mg disodium hydrogen phosphate (dodecanoate), and water for injection per milliliter. The pH was adjusted using 1N hydrochloric acid or sodium hydroxide aqueous solution. The study was conducted at room temperature (25–30°C) and under refrigeration (2–8°C) for different durations. The specific results are shown in Table 2.

[0556] Table 2 Stability Assessment Table

[0557]

[0558] The results showed that compound 27 exhibited poor physical stability at pH < 7.3, undergoing peptide denaturation and precipitation. At pH > 7.4, the solution was clear and showed good stability. Furthermore, HPLC analysis of compound 27 revealed a significant decrease in its content at pH < 7.32, consistent with its state change, indicating that peptide denaturation of compound 27 at pH < 7.4 leads to a decrease in its content.

[0559] Because strongly alkaline preparations are unsuitable for clinical use, the pH of injection solutions is generally not higher than 8.5, and preferably not higher than 8.2. Compound 27 uses a phosphate system with a pH of 7.5.

[0560] Example 17: Selection of formulation excipient dosage for compound 27

[0561] 1. Investigation on the dosage of disodium hydrogen phosphate

[0562] Different phosphate solutions were prepared according to the 1.5 mg / ml specification of compound 27 active pharmaceutical ingredient, and the dosage of disodium hydrogen phosphate was determined with the target pH value of the drug-containing solution being 7.5. The specific results are shown in Table 3.

[0563] Table 3. Investigation of Salt Usage

[0564]

[0565] Conclusion: The dosage of disodium hydrogen phosphate (dodecanoate) is 2.0 mg / ml.

[0566] 2. Investigation into Glycerin Dosage

[0567] Phosphate solutions with different amounts of glycerol were prepared using compound 27 active pharmaceutical ingredient at 1.5 mg / ml, disodium hydrogen phosphate (dodecanoate) at 2.0 mg / ml, and phenol at 5.0 mg / ml. Osmotic pressure was used as the main evaluation index. The specific results are shown in Table 4.

[0568] Table 4. Results of osmotic pressure measurements at different glycerol dosages

[0569]

[0570] The results showed that the osmolality of the solution with a glycerol dosage of 19.7 mg / ml was closest to the normal human osmolality value of 285-310 mOsmol / kg, and the glycerol dosage of 19.7 mg / ml was selected.

[0571] 3. Formulation

[0572] According to the formulation, first prepare a solution by mixing the required amounts of disodium hydrogen phosphate, phenol, glycerin, and an appropriate amount of sterile water for injection. Then, add the active pharmaceutical ingredient to the solution, stir to dissolve, and then add an appropriate amount of sterile water for injection to reach the target volume. The pH of the formulation solution can be adjusted using 1N HCl and 1N NaOH solutions. Finally, sterilize the formulation solution by filtering it through a 0.22-micron filter membrane. This preparation method is applicable to the formulations of all compounds of this invention.

[0573] 4. Pharmaceutical formulation

[0574] The formulations of compound 27 are shown in Tables 5 and 6.

[0575] Table 5 Formulation 1

[0576]

[0577] Table 6 Formulation 2

[0578]

[0579] Example 18: Formulation of Compound 220

[0580] The formulation of compound 220 is shown in Table 7-9.

[0581] Table 7 Formulation 3

[0582]

[0583] Table 8 Formulation Formula 4

[0584]

[0585] Table 9 Formulation Formula 5

[0586]

[0587] Example 19 Formulation of Compound 221

[0588] The formulations of compound 221 are shown in Tables 10 and 11.

[0589] Table 10 Formulation Prescriptions (VI)

[0590]

[0591] Table 11 Formulation Formula 7

[0592]

[0593] Example 20 Formulation stability study

[0594] RP-HPLC detection method: Mobile phase A: 50 mM KH₂PO₄ / K₂HPO₄ aqueous solution (containing 1.203 g K₂HPO₄ and 5.865 g KH₂PO₄ per liter), pH=6.0; B: 100% acetonitrile; gradient: 38-55%, 0-50 min; column: Luna C18, 3 µm (4.6 x 150 mm); flow rate: 0.8 mL / min; detection wavelength: 214 nm

[0595] Content determination: The standard was diluted with water for injection to prepare solutions of different concentrations, and the above-mentioned high performance liquid chromatography method was used for detection. Different peak areas were obtained for the standard at different concentrations. Then, a standard curve was generated using Excel software to obtain the calculation formula.

[0596] Sample content calculation: Take a sample and test it using the above-mentioned high performance liquid chromatography method. Substitute the obtained peak area into the calculation formula of the standard curve to obtain the sample content value.

[0597] The content of related substances is calculated as the proportion of the impurity peak area to the total integral area.

[0598] The formulation samples were placed under long-term conditions (temperature 5℃±3℃) for 6 months, and samples were taken for testing at the end of the 1st, 2nd, 3rd and 6th months respectively (Table 12).

[0599] Table 12

[0600]

[0601] The formulation samples were placed under accelerated conditions (25℃±2℃, 60%±5%) for 6 months, and samples were taken for testing at the end of the 1st, 3rd and 6th months respectively (Table 13).

[0602] Table 13

[0603]

[0604] The injection solutions of prescriptions one through seven above all performed within the compliant range under long-term conditions for 6 months. The injection solution of prescription one, however, remained within the compliant range for 25 months. o Under accelerated conditions, all evaluation indicators were within compliance ranges within 6 months.

[0605] Example 21

[0606] The db / db mice were housed under the same conditions as in Example 7. Male db / db mice (8 weeks old) were acclimatized to the experimental environment for one week. Baseline blood glucose and body weight were recorded for the first three days of the experiment. Based on these three-day blood glucose and body weight data, the mice were randomly divided into four groups of eight mice each. Every 72 hours, the mice were subcutaneously injected with either a blank injection solution (each milliliter containing: disodium hydrogen phosphate (dodecanoate) 2.0 mg, glycerol 18.0 mg, phenol 5.0 mg, and sterile water for injection 974.4 mg; control group) or peptide compounds 27, 220, and 221 (8 nmol / kg, prepared according to prescriptions 2, 3, and 6, respectively), repeated 10 times. Blood samples were collected on day 31 to measure HbA1c (%). Results: Control group 6.9±0.5, compound 27 group 5.6±0.4, compound 220 group 5.3±0.2, compound 221 group 5.1±0.1. There were statistically significant differences between the treated groups and the control group. Therefore, formulations of compounds 27, 220, and 221 show promise as treatments for diabetes.

Claims

1. A pharmaceutical formulation comprising a peptide compound of formula (I) or a pharmaceutically acceptable salt thereof. Y-aib-EGTFTSD-X1-SI-X2-L-X3-X4-EA-X5-X6-LF-X7-X8-WL-X9-AG-X10(I) wherein X1 represents L amino acid, X2 represents amino acid selected from A or aib, X3 represents E amino acid, X4 represents K amino acid, X5 represents V amino acid, X6 represents amino acid selected from K or R, X7 represents I amino acid, X8 represents E amino acid, X9 represents L amino acid, and X10 represents GPSSGAPPP. One or two amino acids containing amino or thiol groups are added to the C-terminus of X10, and the carboxyl group of the C-terminal amino acid is amidated to form a C-terminal amide. The amino acid has the formula... (II) or (III), The wavy line represents the connection point to the adjacent group, n1 is an integer from 1 to 7, and when II or III is a C-terminal amino acid, its carboxyl group is CONH2. The amino acid containing a side-chain amino or thiol group added to the C-terminus of X10 has its side-chain amino or thiol group modified by a long-acting group having the structure of formula (IV): O1-O2-O3-O4-O5-O6-O7-O8-(IV), Where O1 represents the structure of formula (V) or (VI): (V) or (VI) Where n² is an integer between 6 and 24; The wavy line represents the linking point of an amino group to an adjacent group, and O2-O3-O4-O5-O6-O7-O8- represents a linker, wherein each of O2 to O8 is independently represented by any one of the following amino acid residues or long chain structures: α-Glu, γ-Glu, α-Asp, β-Asp, α-hGlu, δ-hGlu, Gly, Ala, β-Ala, GABA or PEG2, or one or more of O2 to O8 are absent, provided that at least two of O2 to O8 are present.

2. The pharmaceutical preparation according to claim 1, wherein the amino acid containing a side-chain amino group is lysine, and the amino acid containing a side-chain thiol group is cysteine.

3. The pharmaceutical preparation according to claim 1, wherein n2 is an integer from 10 to 24.

4. The pharmaceutical preparation according to claim 1, wherein n2 is an integer from 16 to 22.

5. The pharmaceutical formulation according to claim 1, wherein O2 to O8 contains at least one negatively charged portion.

6. The pharmaceutical formulation according to any one of claims 1-5, wherein O2-O3-O4-O5-O6-O7-O8- represents a linker selected from the group consisting of γGlu-PEG2-γGlu-, γGlu-PEG2-2×γGlu-, γGlu-PEG2-, γGlu-2×PEG2-, γGlu-3×PEG2-, γGlu-PEG2-γGlu-PEG2, γGlu-2×PEG2-γGlu-, γGlu-2×PEG2-2×γGlu-, 2×γGlu-, 2×γGlu-PEG2-γGlu-, 2×γGlu-PEG2-γGlu-, 2×γGlu-2×PEG2-, 2×γGlu-2×PEG2-γGlu, and 2×γGlu-2×PEG2-2×γGlu-.

7. The pharmaceutical formulation according to claim 2, wherein the lysine side chain amino-conjugated long-acting group has a structure of formula (VIII) or formula (IX): (VIII); (XI); and (XII). The wavy line represents the connection point to the adjacent amino acid residue.

8. The pharmaceutical formulation according to any one of claims 1-5, wherein the amino acid containing a side-chain thiol added to the C-terminus of X10 has its side-chain thiol modified by a long-acting group of formula (IV), wherein the connection relationship between the side-chain thiol of the amino acid containing the side-chain thiol and the long-acting group is: side-chain thiol of the amino acid containing the side-chain thiol - thiol reactive group - optional linking group L - long-acting group, wherein the side-chain thiol of the amino acid containing the side-chain thiol is connected to one end of the linking group L via a Michael reaction acceptor or a thiol reactive group.

9. The pharmaceutical formulation according to claim 8, wherein the other end of the linking group L is further covalently linked to the long-acting group of formula (IV) via an amino or carboxyl group.

10. The pharmaceutical formulation according to claim 8, wherein the linking group L is selected from: -NH-(CH2) n5 -(CH2CH2O) n6 -(CH2) n7 -,-NH-(CH2) n5 -(CH2CH2O) n6 -(CH2) n7 -NH-, -NH-(CH2) n5 -(CH2CH2O) n6 -(CH2) n7 -CO-, -NH-(CH2) n5 -(CH2CH2O) n6 -(CH2) n7 -NHCO-(CH2) n8 -,-NH-(CH2) n5 -(CH2CH2O) n6 -(CH2) n7 -NHCO-(CH2) n8 -NH- or any combination thereof, where n3, n4, n5, n6, n7, and n8 are each integers from 0 to 10.

11. The pharmaceutical formulation according to claim 8, wherein L is -NH-CH2-(CH2CH2O)3-(CH2)3-NH- or -NH-(CH2). n5 -(CH2CH2O) n6 -(CH2) n7 -NHCO-(CH2) n8 - 12. The pharmaceutical formulation according to any one of claims 1-5, 7, 9-11, wherein said peptide compound is selected from any one of: (1) X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X6 is R, X7 is I, X8 is E, X9 is L, X10 is GPSSGAPPP; (2) X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, X6 is R, X7 is I, X8 is E, X9 is L, X10 is GPSSGAPPP; (3) X1 is L, X2 is A, X3 is E, X4 is K, X5 is V, X6 is K, X7 is I, X8 is E, X9 is L, X10 is GPSSGAPPP; (4) X1 is L, X2 is aib, X3 is E, X4 is K, X5 is V, X6 is K, X7 is I, X8 is E, X9 is L, and X10 is GPSSGAPPP.

13. The pharmaceutical formulation according to claim 12, wherein the C-terminal lysine side chain of X10 is conjugated with a long-acting group.

14. The pharmaceutical formulation according to claim 12, wherein the C-terminal lysine side chain amino-conjugated long-acting group of X10 has the structure of formula (VIII) or formula (IX).

15. The pharmaceutical formulation according to claim 1, wherein the peptide compound is selected from: Compound 27. Compound 29. Compound 220. Compound 221. 。 16. The pharmaceutical formulation according to any one of claims 1-5, 7, 9-11, 13-15, wherein the pharmaceutical formulation further comprises a carrier or excipient selected from one or more of buffer solutions, preservatives, isotonic agents, solubilizers, stabilizers, surfactants, and chelating agents.

17. The pharmaceutical formulation of claim 16, wherein the buffer is selected from one or more of phosphate buffer, hydrogen phosphate buffer, dihydrogen phosphate buffer, acetate buffer, carbonate buffer, bicarbonate buffer, borate buffer, maleate buffer, fumarate buffer, lactate buffer, citrate buffer, sodium barbital buffer, and tris(hydroxymethyl)aminomethane (Tris) buffer.

18. The pharmaceutical preparation according to claim 16, wherein the buffer solution is one or more of disodium hydrogen phosphate, dipotassium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, and sodium acetate solution.

19. The pharmaceutical preparation of claim 16, wherein the buffer solution is a disodium hydrogen phosphate solution, wherein the disodium hydrogen phosphate is anhydrous disodium hydrogen phosphate or disodium hydrogen phosphate hydrate.

20. The pharmaceutical preparation according to claim 16, wherein the preservative is selected from one or more of phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, phenyl nitrite mercury, phenoxyethanol, formaldehyde, chlorobutanol, magnesium chloride, alkyl p-hydroxybenzoates, benzalkonium chloride, benzyl chloride, sodium methyl pyranone, and thimerosal. The magnesium chloride includes magnesium chloride hexahydrate; The alkyl parabens include methylparaben, ethylparaben, propylparaben, and / or butylparaben.

21. The pharmaceutical preparation of claim 16, wherein the preservative is phenol or m-cresol.

22. The pharmaceutical preparation of claim 16, wherein the preservative is phenol.

23. The pharmaceutical formulation of claim 16, wherein the isotonic agent is selected from one or more of glycerol, propylene glycol, monosaccharides, disaccharides, polysaccharides, aldehydes, and sodium chloride.

24. The pharmaceutical preparation according to claim 23, wherein the monosaccharide is selected from one or more of fructose, maltose, galactose, glucose, D-mannose, and sorbitol; the disaccharide is selected from one or more of lactose, sucrose, trehalose, and cellobiose; the polysaccharide is selected from one or more of raffinose, melitriose, maltodextrin, dextran, and starch; and the aldehyde alcohol is selected from one or more of mannitol, xylitol, maltitol, lactitol, xylitol, and sorbitol.

25. The pharmaceutical formulation of claim 16, wherein the isotonic agent is glycerol.

26. The pharmaceutical formulation of claim 16, wherein the isotonic agent is propylene glycol.

27. The pharmaceutical formulation of claim 16, wherein the excipient is a sugar excipient; the sugar excipient is selected from one or more of monosaccharides, disaccharides, polysaccharides, and aldehydes and alcohols.

28. The pharmaceutical formulation according to claim 16, wherein the surfactant is selected from polysorbate, Tween 20 (polyoxyethylene (20) dehydrated sorbitan monolaurate), Tween 40 (polyoxyethylene (20) dehydrated sorbitan monolaurate), Tween 80 (polyoxyethylene (20) dehydrated sorbitan monooleate), Prologne F68 (polyoxyethylene polyoxypropylene block copolymer), PEG (polyethylene glycol), polysorbate 20 or 80, poloxamer 184 or 188, prologne, and one or more other block copolymers.

29. The pharmaceutical formulation according to claim 16, wherein the pH of the pharmaceutical formulation is between 3 and 9.

30. The pharmaceutical formulation according to claim 16, wherein the pH of the pharmaceutical formulation is between 4 and 9.

31. The pharmaceutical preparation according to claim 30, wherein the pharmaceutical preparation has a pH value of pH > 6.

5.

32. The pharmaceutical preparation according to claim 30, wherein the pH of the pharmaceutical preparation is ≥7.

4.

33. The pharmaceutical formulation according to claim 30, wherein the pH value is in the range of 6.5-8.

5.

34. The pharmaceutical formulation according to claim 30, wherein the pH value is in the range of 6.6-8.

0.

35. The pharmaceutical formulation according to claim 30, wherein the pH value is in the range of 7.2-8.

0.

36. The pharmaceutical formulation according to claim 30, wherein the pH value is in the range of 7.4-8.

0.

37. The pharmaceutical formulation according to claim 30, wherein the pH value is in the range of 7.5-7.

6.

38. The pharmaceutical preparation according to claim 16, wherein the dosage form of the pharmaceutical preparation is selected from tablets, sugar tablets, hard and soft gelatin capsules, emulsions, suspensions, suppositories, and injections.

39. The pharmaceutical formulation according to any one of claims 1-5, 7, 9-11, 13-15, wherein the concentration of the peptide compound is at least 0.1 mg / ml.

40. The pharmaceutical formulation according to claim 39, wherein the concentration of the peptide compound is 0.5-20 mg / ml, 0.5-10 mg / ml, 1-8 mg / ml, 1-6 mg / ml, 1-5 mg / ml or 1.5-4 mg / ml.

41. The pharmaceutical formulation according to claim 39, wherein the concentration of the peptide compound is 1.5 mg / ml.

42. The pharmaceutical preparation according to claim 16, wherein the buffer solution is a disodium hydrogen phosphate solution.

43. The pharmaceutical formulation according to claim 42, wherein the concentration of the disodium hydrogen phosphate is at least 0.1 mg / ml.

44. The pharmaceutical preparation according to claim 43, wherein the concentration of the disodium hydrogen phosphate is 0.1-10 mg / ml, 0.3-10 mg / ml, 0.5-10 mg / ml, 0.5-5 mg / ml, 0.5-3 mg / ml or 0.7-2 mg / ml.

45. The pharmaceutical formulation according to claim 16, wherein the isotonic agent is glycerol.

46. ​​The pharmaceutical formulation according to claim 45, wherein the concentration of the glycerol is at least 12 mg / ml.

47. The pharmaceutical formulation according to claim 45, wherein the concentration of glycerol is 15-25 mg / ml, 17-22 mg / ml, or 17-20 mg / ml.

48. The pharmaceutical formulation according to claim 16, wherein the isotonic agent is propylene glycol.

49. The pharmaceutical formulation according to claim 48, wherein the concentration of propylene glycol is at least 8 mg / ml.

50. The pharmaceutical formulation according to claim 48, wherein the concentration of propylene glycol is 10-20 mg / ml, 10-18 mg / ml, 10-16 mg / ml, 12-16 mg / ml, or 13-15 mg / ml.

51. The pharmaceutical preparation according to claim 16, wherein the preservative is phenol.

52. The pharmaceutical preparation according to claim 51, wherein the concentration of phenol is not greater than 5.5 mg / ml or not greater than 5.0 mg / ml.

53. The pharmaceutical formulation according to claim 16, wherein the concentration of the peptide compound is 1.5 mg / ml; the amount of disodium hydrogen phosphate dodecahydrate is 2 mg / ml; the concentration of glycerol is 18 mg / ml or 19.7 mg / ml; and the concentration of phenol is 5 mg / ml.

54. The pharmaceutical formulation according to claim 16, wherein the concentration of the peptide compound is 1.5 mg / ml; the amount of disodium hydrogen phosphate dodecahydrate is 2.84 mg / ml; the concentration of propylene glycol is 14 mg / ml; and the concentration of phenol is 5 mg / ml.

55. Use of the peptide pharmaceutical formulation according to any one of claims 1-54 in the preparation of a medicament for the treatment or prevention of the following diseases or conditions: impaired glucose tolerance (IGT), hyperglycemia, type 1 diabetes, type 2 diabetes, obesity, metabolic syndrome, and neurodegenerative diseases.

56. The use of claim 55, wherein the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, Huntington's disease, ataxia, Kennedy disease, myotonic dystrophy, multi-systemic atrophy, amyotrophic lateral sclerosis, primary lateral sclerosis, spinal muscular atrophy, prion-associated diseases, multiple sclerosis, capillary aplasia, Batten disease, corticobasal degeneration, subacute combined degeneration of the spinal cord, tabes dorsalis, Tay-Sachs disease, toxic encephalopathy, infantile Refsum disease, Refsum disease, neuroacanthosis, Niemann-Pick disease, Lyme disease. Diseases including Machado-Joseph disease, Sandhoff disease, Shy-Drager syndrome, wobbly hedgehog syndrome, cerebral β-amyloidosis, retinal ganglion cell degeneration in glaucoma, synucleinopathies, tauopathies, frontotemporal lobe degeneration (FTLD), dementia, Cadasil syndrome, hereditary cerebral hemorrhage with amyloidosis, Alexander disease, seipinopathies, familial amyloid neuropathy, senile systemic amyloidosis, serpinopathies, AL amyloidosis, AH amyloidosis, AA amyloidosis, and intraaortic amyloidosis. (medialamyloidosis), ApoAI amyloidosis, ApoAII amyloidosis, ApoAIV amyloidosis, Finnish familial amyloidosis (FAF), lysozyme amyloidosis, fibrinogen amyloidosis, or dialysis amyloidosis. The ataxia mentioned includes spinocerebellar ataxia; The prion-related diseases mentioned include Creutzfeldt-Jacob disease.

57. Use of the peptide pharmaceutical formulation according to any one of claims 1-54 in the preparation of a medicament for delaying or preventing the progression of type 2 diabetes, delaying the progression from impaired glucose tolerance to type 2 diabetes, or delaying the progression from type 2 diabetes to insulin-dependent diabetes.

58. Use of the peptide pharmaceutical formulation according to any one of claims 1-54 in the preparation of a medicament for regulating appetite, inducing satiety, reducing food intake, increasing energy expenditure, or preventing weight rebound after successful weight loss.

59. Use of the peptide pharmaceutical formulation according to any one of claims 1-54 in the preparation of a medicament for the treatment or prevention of the following diseases or conditions: treatment of atherosclerosis, hypertension, coronary heart disease, β-blocker poisoning; non-alcoholic fatty liver disease (NAFLD); for the inhibition of gastrointestinal motility; treatment of inclusion body myositis / myopathy, cataracts, retinitis pigmentosa with rhodopsin mutations, medullary thyroid carcinoma, atrial amyloidosis, pituitary prolactinoma, hereditary lattice corneal dystrophy, cutaneous lichen amyloidosis, Mallory bodies, corneal lactoferrin amyloidosis, pulmonary alveolar proteinosis, pindborg tumor amyloidosis, cystic fibrosis, sickle cell disease or critical illness myopathy (CIM), and skeletal-related conditions; The non-alcoholic fatty liver disease includes simple fatty liver (SFL), non-alcoholic steatohepatitis (NASH), and related cirrhosis.

60. The use according to claim 55 or 58, wherein said use is the preparation of a medicament for weight loss.

61. Use of the pharmaceutical preparation according to any one of claims 1-54 in the preparation of a medicament for lowering blood lipids.

62. According to the use of claim 61, the lipid-lowering method is to lower lipid components selected from the following: cholesterol, triglycerides, free fatty acids, and low-density lipoprotein cholesterol.

63. Use of the pharmaceutical preparation according to any one of claims 1-54 in the preparation of a medicament for lowering blood sugar or treating diabetes.

64. A method for preparing a pharmaceutical formulation according to any one of claims 1-54, wherein the preparation method comprises the following steps: (1) Dissolve the buffer, isotonic agent and optional preservative and surfactant in an appropriate amount of sterile water for injection to obtain a solution; (2) Add the peptide compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-54 and adjust to the desired pH range; (3) Add an appropriate amount of sterile water for injection to bring the formulation solution to the target volume; (4) Use a 0.22-micron filter membrane to sterilize the preparation solution.

65. The preparation method according to claim 64, wherein the buffer is disodium hydrogen phosphate or disodium hydrogen phosphate hydrate.

66. The preparation method according to claim 64, wherein the preservative is phenol or m-cresol.

67. The preparation method according to claim 64, wherein the preservative is phenol.

68. The preparation method according to claim 64, wherein the isotonic agent is glycerol or propylene glycol.

69. The preparation method according to claim 64, wherein the isotonic agent is glycerol.

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