Long-acting peptide tyrosine (PYY) analog and method of use

Abstract

The problem that this invention aims to solve is to provide a compound that is a peptide tyrosine (PYY) analog and a method for preparing the same. [Solution] The present invention relates to an isolated polypeptide comprising the amino acid sequence of SEQ ID NO: 90, or a pharmaceutically acceptable salt thereof.

Description

Technical Field

[0001] Related Application This application claims the benefit of U.S. Provisional Patent Application No. 63 / 011,649 filed on April 17, 2020, and U.S. Provisional Patent Application No. 63 / 076,459 filed on September 10, 2020, the disclosures of which are incorporated herein by reference in their entireties.

[0002] The present invention relates to compounds that are peptide tyrosine tyrosine (PYY) analogs and methods for their preparation. The present invention also provides pharmaceutically acceptable compositions comprising the compounds of the invention, and methods of using said compositions in the treatment of various disorders.

Background Art

[0003] Peptide YY (PYY), also known as peptide tyrosine tyrosine, is a peptide encoded by the PYY gene in humans. PYY is a short (36 amino acid) peptide released from cells of the ileum and colon in response to feeding. In the blood, intestine, and other peripheral compartments, PYY acts to reduce appetite; similarly, when injected directly into the central nervous system, PYY is also anorexigenic, i.e., it reduces appetite.

Summary of the Invention

[0004] It has now been found that the compounds of the present disclosure, and pharmaceutically acceptable compositions thereof, are effective as PYY analogs. Such compounds have the following general formula: The amino acid sequence of SEQ ID NO: 90: X0PX2PX4X5PX7X8DX 10 SPX 13 X 14 X 15 X 16 RX 18 X 19 X 20 DX 22 X 23 HX 25 LX 27 WLTRX 32 RX 34Having an isolated polypeptide containing -(OH / NH2) (SEQ ID NO: 90), or a pharmaceutically acceptable salt thereof. (In the formula, each variable is as defined and described herein.)

[0005] Examples of such compounds are provided in Table 3 of this specification.

[0006] The compounds of the present invention have a long disappearance half-life (t 1 / 2 It is designed to achieve ) and is therefore described herein as a “long-acting” PYY analog.

[0007] The compounds of the present invention and their pharmaceutically acceptable compositions are useful for treating a variety of diseases, disorders, or conditions associated with the PYY receptor. Such diseases, disorders, or conditions include metabolic diseases or disorders such as type 2 diabetes, obesity, and the need to achieve weight loss. In certain embodiments, the present invention also relates to a method for treating non-alcoholic fatty liver disease (NAFLD) and / or non-alcoholic steatohepatitis (NASH). [Brief explanation of the drawing]

[0008] [Figure 1] This shows the mean dose reduction (%) from baseline and vehicle control (ΔΔ%) of the long-acting PYY analog in combination with the long-acting GLP-1 analog. [Figure 2] The mean HbA1c (%) from baseline and vehicle control (ΔΔ%) for long-acting PYY analogs combined with long-acting GLP-1 analogs is shown. [Figure 3] This shows a cross-sectional view of a typical osmotic minipump used for drug delivery. [Figure 4] This shows the change in plasma concentration of A13 after bolus injection or intravenous infusion. [Figure 5] This shows the change in plasma concentration of A24 after bolus injection or intravenous infusion. [Figure 6]This graph shows weight loss in a diet-induced obesity rat model, highlighting that the combination of PYY analog peptide A13 and a long-acting GLP-1 receptor agonist is significantly more effective and potent than the industry benchmarks of exenatide or semaglutide alone. [Figure 7] This graph demonstrates the antidiabetic effect in Zucker diabetic obese (ZDF) rats with type 2 diabetes, highlighting that the combination of PYY analog peptide A13 and a long-acting GLP-1 receptor agonist is significantly more effective and potent than tilzepatide and semaglutide alone. [Figure 8] This panel of graphs shows the changes in food intake (ΔΔg) (left panel) and corresponding changes in body weight (ΔΔg, right panel) over different treatment periods at different doses in a diet-induced obesity rat model. The upper panel corresponds to repeated administration of PYY analog A24 alone, and the lower panel corresponds to combination administration of PYY analog A24 and a long-acting GLP-1 receptor agonist. [Figure 9] Figure 8 shows the Y-dimensional data replotted on the X and Y axes. The change in body weight (ΔΔg, Y-axis) is plotted as a function of the change in food intake (ΔΔg, X-axis). The good fit of the relationship (R20.82) indicates that the impact on food intake highly predicts the impact on body weight. The slope of this relationship shows that in this animal model, a change (increase or decrease) in cumulative food intake over a given period accounts for approximately 56% of the corresponding change in body weight. [Modes for carrying out the invention]

[0009] General description of a specific embodiment of the present invention The compounds and pharmaceutical compositions of the present invention are useful as agonists of PYY receptors, particularly as agonists of human PYY receptors including NPY1R, NPY2R, NPY4R, and NPY5R. The present invention also relates to methods for producing and using such compounds, i.e., PYY analog polypeptides. The compounds of the present invention are long-acting PYY analogs. These PYY analog polypeptides are particularly useful in methods for treating metabolic diseases or disorders such as type 2 diabetes and obesity, and in methods for weight loss. In certain embodiments, the present invention also relates to methods for treating non-alcoholic fatty liver disease (NAFLD) and / or non-alcoholic steatohepatitis (NASH).

[0010] In a particular embodiment, the present disclosure relates to the amino acid sequence of SEQ ID NO: 90: X0PX2PX4X5PX7X8DX 10 SPX 13 X 14 X 15 X 16 RX 18 X 19 X 20 DX 22 X 23 HX 25 LX 27 WLTRX 32 RX 34 This provides an isolated polypeptide containing -(OH / NH2) (SEQ ID NO: 90), or a pharmaceutically acceptable salt thereof.

[0011] definition It should be understood that the terms used herein are for the purpose of describing specific embodiments and are not intended to limit them. As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless otherwise clearly indicated by the context. Thus, for example, a reference to “solvent” includes combinations of two or more such solvents; a reference to “peptide” includes one or more peptides or mixtures of peptides; a reference to “drug” includes one or more drugs; a reference to “osmotic delivery device” includes one or more osmotic delivery devices, and so on. Unless specifically stated or evident from the context, the term “or” as used in the present invention is understood to be inclusive and encompasses both “or” and “and.”

[0012] Unless otherwise stated or made clear from the context, the term “about” as used herein is understood to mean within the normal range of acceptance in the art, for example, within two standard deviations of the mean. “About” may be understood to mean within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise made clear from the context, all numerical values ​​provided herein are modified by the term “about.”

[0013] Unless otherwise specifically stated or evident from the context, the term “substantially” as used herein is understood to mean a narrow range of variation in the art or otherwise within the normal acceptable range. Substantially, it can be understood as within 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, 0.01%, or 0.001% of the stated value.

[0014] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which the present invention pertains. Other methods and materials similar to or equivalent to those described herein may be used in the practice of the present invention, but preferred materials and methods are described herein.

[0015] In describing and claiming the present invention, the following terms will be used in accordance with the definitions set forth below.

[0016] The terms “drug,” “therapeutic agent,” and “beneficial agent” refer to any therapeutically active substance that is interchangeable, delivered to a target, and produces a desired beneficial effect. In one embodiment of the present invention, the drug is a polypeptide. In another embodiment of the present invention, the drug is a small molecule, such as a hormone, like an androgen or estrogen. The devices and methods of the present invention are particularly well suited for the delivery of proteins, small molecules, and combinations thereof.

[0017] The terms “peptide,” “polypeptide,” and “protein” are used interchangeably herein and typically refer to molecules comprising chains of two or more amino acids (e.g., most typically L-amino acids, but also including, for example, D-amino acids, modified amino acids, amino acid analogs, and amino acid mimes).

[0018] In some embodiments, naturally occurring L-amino acids are represented by the conventional three-letter or single-letter uppercase amino acid notation in Table 1. In other embodiments, both naturally occurring L-amino acids and D-amino acids are represented by the conventional three-letter or single-letter uppercase amino acid notation in Table 1. In yet another embodiment, D-amino acids correspond to the single-letter lowercase amino acid notation in Table 2, namely a, l, m, f, w, k, q, e, s, p, v, i, c, y, h, r, n, d, and t. [Table 1] [Table 2]

[0019] Peptides may be naturally occurring, synthetically produced, or recombinantly expressed. Peptides may also contain additional groups that modify the amino acid chain, such as functional groups added by post-translational modifications. Examples of post-translational modifications include, but are not limited to, acetylation, alkylation (including methylation), biotinylation, glutamylation, glycylation, glycosylation, isoprenylation, lipoylation, phosphopantetheinylation, phosphorylation, selenization, and C-terminal amidation. The term peptide also includes peptides with amino-terminal and / or carboxyl-terminal modifications. Modifications of terminal amino groups include, but are not limited to, desamino modification, N-lower alkyl modification, N-di-lower alkyl modification, and N-acyl modification. Modifications of terminal carboxyl groups include, but are not limited to, amide modification, lower alkylamide modification, dialkylamide modification, and lower alkyl ester modification (e.g., lower alkyls are C1-C4 alkyls). The term peptide also includes modifications of amino acids between the amino and carboxyl terms, such as, but not limited to, those described above. In one embodiment, a peptide may be modified by the addition of a small molecule drug.

[0020] The terminal amino acid at one end of a peptide chain typically has a free amino group (i.e., the amino terminus). The terminal amino acid at the other end of the chain typically has a free carboxyl group (i.e., the carboxy terminus). Typically, the amino acids that make up a peptide are numbered sequentially, starting from the amino terminus and increasing towards the carboxy terminus of the peptide.

[0021] As used herein, the term “amino acid residue” refers to an amino acid incorporated into a peptide by an amide bond or an amide bond mimetic.

[0022] As used herein, the term “insulin-secreting” typically refers to the ability of a compound, such as a peptide, to stimulate or influence the production and / or activity of insulin (e.g., insulin-secreting hormones). Such compounds typically stimulate or otherwise influence the secretion or biosynthesis of insulin in a subject. Thus, an “insulin-secreting peptide” is an amino acid-containing molecule that can stimulate or otherwise influence the secretion or biosynthesis of insulin.

[0023] As used herein, the term “acylation” means that, with respect to the disclosed polypeptide, the disclosed polypeptide is optionally substituted with one or more lipophilic substituents via spacers, and “lipophilic substituent” and “spacer” are defined herein. Certain lipophilic substituents may optionally bind to albumin via spacers, conferring affinity to albumin to the resulting acylated polypeptide. The extent to which lipophilic substituents optionally bind to albumin via spacers and confer affinity to albumin to the resulting acylated polypeptide varies and depends on many factors. Many factors include the identity of the lipophilic substituents, the optional spacers, the polypeptide, and the covalent binding sites to the polypeptide.

[0024] The terms “linear” or “linear polypeptide” refer to “non-acylated” polypeptides, in other words, disclosed PYY analog polypeptides that do not have lipophilic substituents via spacers, respectively, where “lipophilic substituents” and “spacers” are defined herein.

[0025] As used herein, the terms “conjugate” or “conjugate polypeptide” refer to “acylated” polypeptides, in other words, disclosed PYY analog polypeptides each having one or more lipophilic substituents via spacers, respectively.

[0026] As used herein, “pharmaceutically acceptable salt” means a derivative of a disclosed polypeptide, wherein the parent polypeptide is modified by converting any existing acidic or base moiety to the form of a salt. A list of preferred salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977), which are incorporated herein by reference in their entirety.

[0027] As used herein, the term “vehicle” refers to a medium used to carry a compound, such as a drug or drug-containing particles. The vehicle of the present invention typically comprises components such as a polymer and a solvent. The suspension vehicle of the present invention typically comprises a solvent and a polymer used to prepare a suspension formulation further comprising a drug particle formulation.

[0028] As used herein, the term “phase separation” refers to the formation of multiple phases (e.g., a liquid phase and a gel phase) in a suspension vehicle, such as when the suspension vehicle comes into contact with an aqueous environment. In some embodiments of the present invention, the suspension vehicle is formulated to exhibit phase separation when it comes into contact with an aqueous environment containing less than approximately 10% water.

[0029] As used herein, the term “single-phase” refers to a homogeneous system of solids, semi-solids, or liquids that is physically and chemically uniform throughout.

[0030] As used herein, the term "dispersion" refers to dissolving, dispersing, suspending, or otherwise distributing a compound, such as a drug particle formulation, in a suspension vehicle.

[0031] As used herein, the phrase “chemically stable” means that degradation products generated over a period defined by chemical pathways such as deamidation (usually by hydrolysis), aggregation, or oxidation are formed in the formulation at an acceptable percentage.

[0032] As used herein, the phrase “physically stable” means that aggregates (e.g., dimers and other high molecular weight products) are formed in the formulation at an acceptable percentage. Furthermore, a physically stable formulation does not change its physical state, for example, from liquid to solid, or from amorphous to crystalline.

[0033] As used herein, the term "viscosity" typically refers to a value determined essentially by the ratio of shear stress to shear rate (see, for example, Considine, DM & Considine, GD, Encyclopedia of Chemistry, 4th Edition, Van Nostrand, Reinhold, NY, 1984): F / A=μ*V / L (Equation 1) In the formula, F / A = shear stress (force per unit area). μ = a proportionality constant (viscosity), and V / L = rate per unit thickness (shear rate).

[0034] From this relationship, the ratio of shear stress to shear rate defines viscosity. Measurements of shear stress and shear rate are typically determined using parallel plate rheometry performed under selected conditions (e.g., a temperature of approximately 37°C). Other methods for determining viscosity include measuring kinematic viscosity using a viscometer, such as a Cannon-Fenske viscometer, an Ubbelohde viscometer for Cannon-Fenske opaque solutions, or an Ostwald viscometer. Generally, the suspension vehicles of the present invention have a viscosity sufficient to prevent the suspended particulate formulations therein from settling during storage and to be used in delivery methods, such as implantable drug delivery devices.

[0035] As used herein, the term "non-aqueous" typically refers to the total water content of a suspension formulation, for example, about 10% by weight or less, e.g., about 7% by weight or less, about 5% by weight or less, and / or less than about 4% by weight. Furthermore, the particulate formulations of the present invention contain less than about 10% by weight, e.g., less than 5% by weight of residual water.

[0036] As used herein, the term “subject” refers to any member of the subphylum Chordata, including but not limited to humans, as well as other primates, including rhesus macaques and other monkey species, and non-human primates such as chimpanzees and other great ape species; domesticated animals such as cattle, sheep, pigs, goats and horses; domesticated mammals such as dogs and cats; laboratory animals, including rodents such as mice, rats and guinea pigs; and birds, including domesticated birds, wild birds and game birds, such as chickens, turkeys and other poultry, ducks and geese. The term does not indicate a specific age or sex; it is therefore intended to encompass both adults and neonates.

[0037] As used herein, the terms “to treat,” “to treat,” and “to treat” mean to reverse, alleviate, delay the onset of, or inhibit the progression of a disease or disorder or one or more of its symptoms described herein. In some embodiments, treatment may be administered after the onset of one or more symptoms. In other embodiments, treatment may be administered even in the absence of symptoms. For example, treatment may be administered to individuals who are highly susceptible to the symptoms before the onset of symptoms (e.g., in light of the history of the symptoms and / or in light of genetic or other susceptibility factors). After the symptoms have subsided, treatment may be continued, for example, to prevent or delay their recurrence.

[0038] As used herein, the term “osmotic delivery device” typically refers to a device used for the delivery of a drug (e.g., a disclosed PYY analog polypeptide) to a subject, the device comprising a reservoir (e.g., made from a titanium alloy) having a lumen containing a suspension formulation comprising the drug (e.g., a disclosed PYY analog polypeptide) and an osmotic agent formulation. A piston assembly located within the lumen separates the suspension formulation from the osmotic agent formulation. A semipermeable membrane is located at the first distal end of the reservoir adjacent to the osmotic agent formulation, and a diffusion moderator (defining the delivery orifice through which the suspension formulation passes when exiting the device) is located at the second distal end of the reservoir, adjacent to the suspension formulation. Typically, the osmotic delivery device is implanted in a subject, for example, subdermally or subcutaneously (e.g., medial, lateral, or posterior upper arm and abdominal region). An exemplary osmotic delivery device is the DUROS® (ALZA Corporation, Mountain View, Calif.) delivery device. Examples of terms synonymous with "osmotic delivery device" include, but are not limited to, "osmotic drug delivery device," "osmotic drug delivery system," "osmotic device," "osmotic delivery device," "osmotic delivery system," "osmotic pump," "implantable drug delivery device," "drug delivery system," "drug delivery device," "implantable osmotic pump," "implantable drug delivery system," and "implantable delivery system." Other terms for "osmotic delivery device" are known in the art.

[0039] As used herein, the term “continuous delivery” typically refers to the substantially continuous release of a drug from an osmotic delivery device to tissues near the implantation site, such as subdermal and subcutaneous tissues. For example, an osmotic delivery device releases a drug at an essentially predetermined rate based on the principle of osmosis. Extracellular fluid enters directly into the osmotic engine of the osmotic delivery device through a semipermeable membrane, and this osmotic engine expands, moving a piston at a slow and constant rate. The movement of the piston forces the drug formulation to be released through the orifice of the diffusion moderator. Thus, the release of a drug from an osmotic delivery device occurs at a slow, controlled, and constant rate.

[0040] As used herein, the term “substantially steady-state delivery” typically refers to the delivery of a drug at or near a target concentration over a defined period, where the amount of drug delivered from the osmotic delivery device is substantially zero-order delivery. Substantially zero-order delivery of an activator (e.g., the disclosed PYY analog polypeptide) means that the rate of drug delivery is constant and independent of the drug available in the delivery system. For example, if zero-order delivery is determined by a standard method (e.g., linear regression), and the rate of drug delivery is graphed against time, then if the line fits the data, the line has a slope of approximately zero.

[0041] As used herein, the term “drug half-life” refers to the time it takes for a drug to be eliminated from the plasma to half its concentration. Drug half-life is typically measured by monitoring how the drug breaks down when administered by injection or intravenously. Drugs are typically detected using, for example, radioimmunoassays (RIA), chromatography, electrochemiluminescence (ECL) assays, enzyme-linked immunosorbent assays (ELISA), or immunoenzyme sandwich assays (IEMA).

[0042] The terms "μg," "mcg," and "ug" are understood to mean "microgram." Similarly, the terms "μl" and "uL" are understood to mean "microliter," and the terms "μM" and "uM" are understood to mean "micromoles."

[0043] The term "serum" is intended to mean blood products from which a substance can be detected. Therefore, the term serum includes at least whole blood, serum, and plasma. For example, "the amount of [substance] in the serum of the subject" would encompass "the amount of [substance] in the plasma of the subject."

[0044] Baseline is defined as the last evaluation on the day of initial placement of the osmotic delivery device (containing drug or placebo) or the day before.

[0045] Peptide YY (PYY) is a 36-amino acid peptide amide with the amino acid sequence (YPIKPEAPGEDASPEELNRYYASLRHYLNLVTRQRY-NH2), which is sequence number 800. PYY inhibits intestinal motility and blood flow (Laburthe, M., Trends Endocrinol Metab. 1(3):168-74 (1990)), mediates intestinal secretion (Cox, HM, et al., Br J Pharmacol 101(2):247-52 (1990); Playford, RJ, et al., Lancet 335(8705):1555-7 (1990)), and stimulates net absorption (MacFayden, RJ, et al., Neuropeptides 7(3):219-27 (1986)). Two major in vivo variants, PYY(1-36) and PYY(3-36), have been identified (e.g., Eberlein, GA, et al., Peptides 10(4), 797-803). (1989). The sequence of PYY, as well as its peptide analogs and peptide derivatives, are known in the art (e.g., U.S. Patents 5,574,010 and 5,552,520).

[0046] Exemplary compound: PYY analog polypeptide Certain disclosed PYY analog polypeptides, including those listed in Table 3 below, exhibit one or more of the following: superior solubility, stability, bioavailability, biological activity and specificity, and longer half-lives than endogenous PYY and known PYY analogs. Certain disclosed PYY analog polypeptides have been developed to accommodate less frequent administration than known PYY analogs. Certain disclosed PYY analog polypeptides have been developed for weekly or monthly injection administration. Certain disclosed PYY analog polypeptides have been developed for administration via implantation of a delivery device containing the PYY analog polypeptide, the delivery device containing doses of the PYY analog polypeptide for up to 3 months, 6 months, 9 months, 1 year, 18 months, or 2 years.

[0047] In some embodiments, the isolated polypeptides of this disclosure include an amino acid sequence selected from the group consisting of the following peptides listed in Table 3: [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6] [Table 3-7] [Table 3-8] [Table 3-9] [Table 3-10] [Table 3-11] [Table 3-12] [Table 3-13] [Table 3-14]

[0048] Table 4 below provides the structural representation of a specific peptide from Table 3: [Table 4-1] [Table 4-2] [Table 4-3]

[0049] [ka] As used herein, dpeg represents -COCH2O(CH2)2O(CH2)2NH-; and dpeg-dpeg represents -COCH2O(CH2)2O(CH2)2NH-COCH2O(CH2)2O(CH2)2NH-; • Carboxy-terminal amino acids, i.e., -F 34 F is shown as -(NH2). 34 This is represented as -NH-CH(CH2Ph)-CONH2.

[0050] In some embodiments, the isolated polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 78, or a pharmaceutically acceptable salt thereof.

[0051] In some embodiments, the isolated polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 13, 24, 42, and 43, or a pharmaceutically acceptable salt thereof.

[0052] In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 13 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 24 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 42 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 43 or a pharmaceutically acceptable salt thereof.

[0053] In some embodiments, the present invention provides compounds shown in Table 3 above, or pharmaceutically acceptable salts thereof. In some embodiments, the present invention provides isolated polypeptides comprising amino acid sequences selected from the group consisting of SEQ ID NOs: 1 to 78, or pharmaceutically acceptable salts thereof. In some embodiments, the pharmaceutically acceptable salt is an acetate salt. In some embodiments, the pharmaceutically acceptable salt is a trifluoroacetic acid (TFA) salt. In some embodiments, the pharmaceutically acceptable salt is a hydrochloric acid (HCl) salt.

[0054] In one embodiment, the compound is an isolated polypeptide containing the amino acid sequence of SEQ ID NO: 13 or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is an acetate salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 13. In some embodiments, the compound is a TFA salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 13. In some embodiments, the compound is an HCl salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 13.

[0055] In one embodiment, the compound is an isolated polypeptide containing the amino acid sequence of SEQ ID NO: 24 or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is an acetate salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 24. In some embodiments, the compound is a TFA salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 24. In some embodiments, the compound is an HCl salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 24.

[0056] In one embodiment, the compound is an isolated polypeptide containing the amino acid sequence of SEQ ID NO: 42 or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is an acetate salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 42. In some embodiments, the compound is a TFA salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 42. In some embodiments, the compound is an HCl salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 42.

[0057] In one embodiment, the compound is an isolated polypeptide containing the amino acid sequence of SEQ ID NO: 43 or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is an acetate salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 43. In some embodiments, the compound is a TFA salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 43. In some embodiments, the compound is an HCl salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 43.

[0058] In some embodiments, the isolated polypeptides of this disclosure include an amino acid sequence selected from the group consisting of the following peptides listed in Table 5: [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4]

[0059] In some embodiments, the isolated polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 101-181, or a pharmaceutically acceptable salt thereof.

[0060] In some embodiments, the isolated polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 113, 124, and 143, or a pharmaceutically acceptable salt thereof.

[0061] In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 113 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 124 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 143 or a pharmaceutically acceptable salt thereof.

[0062] In some embodiments, the present invention provides compounds shown in Table 5 above, or pharmaceutically acceptable salts thereof. In some embodiments, the present invention provides isolated polypeptides comprising amino acid sequences selected from the group consisting of SEQ ID NOs: 101 to 181, or pharmaceutically acceptable salts thereof. In some embodiments, the pharmaceutically acceptable salt is an acetate salt. In some embodiments, the pharmaceutically acceptable salt is a trifluoroacetate (TFA) salt. In some embodiments, the pharmaceutically acceptable salt is a hydrochloric acid (HCl) salt.

[0063] In one embodiment, the compound is an isolated polypeptide containing the amino acid sequence of SEQ ID NO: 113 or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is an acetate salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 113. In some embodiments, the compound is a TFA salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 113. In some embodiments, the compound is an HCl salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 113.

[0064] In one embodiment, the compound is an isolated polypeptide containing the amino acid sequence of SEQ ID NO: 124 or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is an acetate salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 124. In some embodiments, the compound is a TFA salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 124. In some embodiments, the compound is an HCl salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 124.

[0065] In one embodiment, the compound is an isolated polypeptide containing the amino acid sequence of SEQ ID NO: 143 or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is an acetate salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 143. In some embodiments, the compound is a TFA salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 143. In some embodiments, the compound is an HCl salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 143.

[0066] In some embodiments, the isolated polypeptides of this disclosure include an amino acid sequence selected from the group consisting of the following peptides listed in Table 6: [Table 6]

[0067] In some embodiments, the isolated polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 280, and 281, or a pharmaceutically acceptable salt thereof.

[0068] In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 239 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 240 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 241 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 242 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 243 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 244 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 245 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 246 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 247 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 248 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 280 or a pharmaceutically acceptable salt thereof. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 281 or a pharmaceutically acceptable salt thereof.

[0069] In some embodiments, the pharmaceutically acceptable salt is an acetate salt. In some embodiments, the pharmaceutically acceptable salt is a trifluoroacetate (TFA) salt. In some embodiments, the pharmaceutically acceptable salt is a hydrochloric acid (HCl) salt.

[0070] In one embodiment, the compound is an isolated polypeptide containing the amino acid sequence of SEQ ID NO: 242 or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is an acetate salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 242. In some embodiments, the compound is a TFA salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 242. In some embodiments, the compound is an HCl salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 242.

[0071] In one embodiment, the compound is an isolated polypeptide containing the amino acid sequence of SEQ ID NO: 243 or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is an acetate salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 243. In some embodiments, the compound is a TFA salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 243. In some embodiments, the compound is an HCl salt of the isolated polypeptide containing the amino acid sequence of SEQ ID NO: 243.

[0072] Description of Exemplary Embodiments In a particular embodiment, the present disclosure relates to the amino acid sequence of SEQ ID NO: 90: X0PX2PX4X5PX7X8X9X 10 SPX 13 X 14 X 15 X 16 RX 18 X 19 X 20 DX 22 X 23 HX 25 X 26 X 27 WLTRX 32 RX 34 Provides an isolated polypeptide containing -(OH / NH2)(SEQ ID NO: 90), or a pharmaceutically acceptable salt thereof (wherein, X0 is either nonexistent or K; X2 is K; X4 is either E or K; X5 is either A or K; X7 is either G or K; X8 is E, K, or k; X9 is either D or K; X 10 is A or K; X 13 is either E or K; X 14 is either E or K; X 15 is either L or W; X 16 These are D, E, K, N, Q, S, T, α-methylserine, or homoserine; X 18 is either K or Y; X 19 is either K or Y; X 20 is A, D, E, K, k, or Dap; X 22 is A, D, K, or L; X 23 is either K or R; X 25 is either K or Y; X 26 is E, K, or L; X 27 is either K or N; X 32 is either K or Q; X 34 These are F, y, 3-pyridinylalanine, 4-pyridinylalanine, 4-carboxyphenylalanine, 4-fluorophenylalanine, 4-methylphenylalanine, N-methylphenylalanine, homophenylalanine, β-homotyrosine, homotyrosine, or N-methyltyrosine; Here, X0, X2, X4, X5, X7, X8, X 13 , X 20 , X 23 , X 25 , X 27 , or X 32 When K is present, the lysine residue is optionally covalently bonded to the lipophilic substituent via a spacer. Here, X8 or X 20When k, the D-lysine residue is optionally covalently bonded to the lipophilic substituent via a spacer. However, the polypeptide is provided that it optionally contains at least one residue covalently bonded to a lipophilic substituent via a spacer; The polypeptide optionally further comprises lactam crosslinks formed via amide bonds between the lysine side chain and the aspartic acid side chain, or between the lysine side chain and the glutamic acid side chain, wherein the residues forming the lactam crosslinks are X9 and X 13 position or X 16 and X 20 position or X 22 and X 26 (To be in a certain position).

[0073] In some embodiments, X 15 If X is L, 22 It is A.

[0074] In some embodiments, X 16 If X is N, 20 It is not A.

[0075] In some embodiments, X8 is E, X5 is K, and X 20 If it is K, then X5 or X 20 One of the lysine residues is optionally covalently bonded to a lipophilic substituent via a spacer.

[0076] In some embodiments, X8 is K. In some embodiments, the lysine residue at position 8 of the polypeptide sequence is optionally covalently bonded to a lipophilic substituent via a spacer.

[0077] In some embodiments, X 15 It is W.

[0078] In some embodiments, X 34 It is F.

[0079] In some embodiments, the peptide is X9 and X13 further comprises a lactam bridge formed via an amide bond between a lysine side chain and a glutamic acid side chain at the position, respectively. In some embodiments, the peptide comprises X9 and X 13 further comprises a lactam bridge formed via an amide bond between an aspartic acid side chain and a lysine side chain at the position, respectively.

[0080] In some embodiments, the peptide comprises X 16 and X 20 further comprises a lactam bridge formed via an amide bond between a lysine side chain and an aspartic acid side chain at the position, respectively. In some embodiments, the peptide comprises X 16 and X 20 further comprises a lactam bridge formed via an amide bond between a lysine side chain and a glutamic acid side chain at the position, respectively. In some embodiments, the peptide comprises X 16 and X 20 further comprises a lactam bridge formed via an amide bond between an aspartic acid side chain and a lysine side chain at the position, respectively. In some embodiments, the peptide comprises X 16 and X 20 further comprises a lactam bridge formed via an amide bond between a glutamic acid side chain and a lysine side chain at the position, respectively. In some embodiments, the peptide comprises X 22 and X 26 further comprises a lactam bridge formed via an amide bond between a lysine side chain and a glutamic acid side chain at the position, respectively. In some embodiments, the peptide comprises X 22 and X 26 further comprises a lactam bridge formed via an amide bond between an aspartic acid side chain and a lysine side chain at the position, respectively.

[0081] In some embodiments, X0 is absent. In some embodiments, X0 is K. In some embodiments, X0 is K covalently bonded to a lipophilic substituent optionally via a spacer.

[0082] In some embodiments, X2 is K. In some embodiments, X2 is K covalently linked to a lipophilic substituent optionally via a spacer.

[0083] In some embodiments, X4 is E. In some embodiments, X4 is K. In some embodiments, X4 is K covalently linked to a lipophilic substituent optionally via a spacer.

[0084] In some embodiments, X5 is A. In some embodiments, X5 is K. In some embodiments, X5 is K covalently linked to a lipophilic substituent optionally via a spacer.

[0085] In some embodiments, X7 is G. In some embodiments, X7 is K. In some embodiments, X7 is K covalently linked to a lipophilic substituent optionally via a spacer.

[0086] In some embodiments, X8 is E. In some embodiments, X8 is K. In some embodiments, X8 is K covalently linked to a lipophilic substituent optionally via a spacer. In some embodiments, X8 is k. In some embodiments, X8 is k covalently linked to a lipophilic substituent optionally via a spacer.

[0087] As used herein, k refers to D-lysine.

[0088] In some embodiments, X9 is D. In some embodiments, X9 is K.

[0089] In some embodiments, X 10 is A. In some embodiments, X 10 is K. In some embodiments, X 10 is K covalently linked to a lipophilic substituent optionally via a spacer.

[0090] In some embodiments, X 13 In some embodiments, X 13 In some embodiments, X 13 This is K, which is optionally covalently bonded to a lipophilic substituent via a spacer.

[0091] In some embodiments, X 14 In some embodiments, X 14 K is K.

[0092] In some embodiments, X 15 L is. In some embodiments, X 15 It is W.

[0093] In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 Q is Q. In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 X is α-methylserine. In some embodiments, X 16 It is homoserine.

[0094] In some embodiments, X 18 In some embodiments, X 18 Y is Y.

[0095] In some embodiments, X 19 In some embodiments, X 19 Y is Y.

[0096] In some embodiments, X 20 In some embodiments, X 20In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 This is K, optionally covalently bonded to a lipophilic substituent via a spacer. In some embodiments, X 20 is k. In some embodiments, X 20 k is optionally covalently bonded to a lipophilic substituent via a spacer. In some embodiments, X 20 It is a DAP.

[0097] As used herein, Dap refers to diaminopimelic acid.

[0098] In some embodiments, X 22 In some embodiments, X 22 In some embodiments, X 22 In some embodiments, X 22 It is L.

[0099] In some embodiments, X 23 In some embodiments, X 23 This is K, which is optionally covalently bonded to a lipophilic substituent via a spacer. In some embodiments, X 23 This is R.

[0100] In some embodiments, X 25 In some embodiments, X 25 This is K, which is optionally covalently bonded to a lipophilic substituent via a spacer. In some embodiments, X 25 Y is Y.

[0101] In some embodiments, X 26 In some embodiments, X 26 In some embodiments, X 26 It is L.

[0102] In some embodiments, X 27 In some embodiments, X 27 This is K, which is optionally covalently bonded to a lipophilic substituent via a spacer. In some embodiments, X 27 It is N.

[0103] In some embodiments, X 32 In some embodiments, X 32 This is K, which is optionally covalently bonded to a lipophilic substituent via a spacer. In some embodiments, X 32 Q is Q.

[0104] In some embodiments, X 34 In some embodiments, X 34 y is. In some embodiments, X 34 is 3-pyridinylalanine. In some embodiments, X 34 is 4-pyridinylalanine. In some embodiments, X 34 X is 4-carboxyphenylalanine. In some embodiments, X 34 X is 4-fluorophenylalanine. In some embodiments, X 34 X is 4-methylphenylalanine. In some embodiments, X 34 is N-methylphenylalanine. In some embodiments, X 34 X is homophenylalanine. In some embodiments, X 34 is β-homotyrosine. In some embodiments, X 34 is homotyrosine. In some embodiments, X 34 It is N-methyltyrosine.

[0105] As used herein, y refers to D-tyrosine.

[0106] In some embodiments, a carboxy-terminal amino acid, namely X 34 is, -X 34It is -(NH2). In some embodiments, the carboxy-terminal amino acid X 34 is, -X 34 It is -(OH).

[0107] In some embodiments, certain amino acids represented by the consensus sequence of Sequence ID No. 90 include the following:

[0108] In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, optionally. In some embodiments, X5 is A, and X 15 is W. In some embodiments, X5 is A and X 16 is D. In some embodiments, X5 is A and X 16 is K. In some embodiments, X5 is A and X 16 is N. In some embodiments, X5 is A and X 20 is K. In some embodiments, X5 is A and X 20 is E. In some embodiments, X5 is A and X 20 is A. In some embodiments, X5 is A, and X 34 It is F.

[0109] In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X8 is E. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 is W. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 20is K. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 34 It is F.

[0110] In some embodiments, X5 is K and X8 is E. In some embodiments, X5 is K and X 15 is W. In some embodiments, X5 is K and X 16 is N. In some embodiments, X5 is K and X 16 is D. In some embodiments, X5 is K and X 20 is K. In some embodiments, X5 is K and X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X5 is K and X 34 It is F.

[0111] In some embodiments, X8 is E and X 15 is W. In some embodiments, X8 is E and X 16 is D. In some embodiments, X8 is E and X 16 is N. In some embodiments, X8 is E and X 20 is K. In some embodiments, X8 is E and X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X8 is E and X 34 It is F.

[0112] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 is W. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is E. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is A. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 34 It is F.

[0113] In some embodiments, X 15 W is X 16 In some embodiments, X 15 W is X 16 In some embodiments, X 15 W is X 16 In some embodiments, X 15 W is X 20 In some embodiments, X 15 W is X 20 In some embodiments, X 15 W is X 20 In some embodiments, X 15 W is X 34 It is F.

[0114] In some embodiments, X 16 is D, X 20 In some embodiments, X 16 is D, X 34 It is F.

[0115] In some embodiments, X 16 is K and X 20In some embodiments, X 16 is K and X 20 In some embodiments, X 16 is K and X 34 It is F.

[0116] In some embodiments, X 16 N is X 20 In some embodiments, X 16 N is X 20 In some embodiments, X 16 N is X 34 It is F.

[0117] In some embodiments, X 20 is K and X 34 It is F.

[0118] In some embodiments, X 20 E is X 34 It is F.

[0119] In some embodiments, X 20 A is X 34 It is F.

[0120] In some embodiments, certain amino acids represented by the consensus sequence of Sequence ID No. 90 include the following:

[0121] In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, X 15 is W. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16is K. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is E. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is A. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 34 It is F.

[0122] In some embodiments, X5 is A, and X 15 W is X 16 is D. In some embodiments, X5 is A and X 15 W is X 16 is D. In some embodiments, X5 is A and X 15 W is X 16 is K. In some embodiments, X5 is A and X 15 W is X 16 is N. In some embodiments, X5 is A and X 15 W is X 20 is K. In some embodiments, X5 is A and X 15 W is X 20 is E. In some embodiments, X5 is A and X 15 W is X 20 is A. In some embodiments, X5 is A, and X 15 W is X 34 It is F.

[0123] In some embodiments, X5 is A, and X16 D is X 20 is K. In some embodiments, X5 is A and X 16 D is X 34 It is F.

[0124] In some embodiments, X5 is A, and X 16 is K and X 20 is E. In some embodiments, X5 is A and X 16 is K and X 20 is D. In some embodiments, X5 is A and X 16 is K and X 34 It is F.

[0125] In some embodiments, X5 is A, and X 16 N is X 20 is A. In some embodiments, X5 is A, and X 16 N is X 20 is K. In some embodiments, X5 is A and X 16 N is X 34 It is F.

[0126] In some embodiments, X5 is A, and X 20 is K and X 34 It is F.

[0127] In some embodiments, X5 is A, and X 20 E is X 34 It is F.

[0128] In some embodiments, X5 is A, and X 20 A is X 34 It is F.

[0129] In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 15is W. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 16 is N. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 16 is D. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 20 is K. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 34 It is F.

[0130] In some embodiments, X5 is K, X8 is E, and X 15 is W. In some embodiments, X5 is K, X8 is E, and X 16 is N. In some embodiments, X5 is K, X8 is E, and X 16 is D. In some embodiments, X5 is K, X8 is E, and X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X5 is K, X8 is E, and X 34 It is F.

[0131] In some embodiments, X8 is E and X 15 W is X 16 is N. In some embodiments, X8 is E and X 15 W is X 20 is K. In some embodiments, X8 is E and X 15 W is X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X8 is E and X 15 W is X 34 It is F.

[0132] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 16 is D. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 16 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 16 is N. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 20 is E. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 20 is A. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 34 It is F.

[0133] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D, X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D, X 34 It is F.

[0134] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K, and X 20is E. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K, and X 20 is D. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K, and X 34 It is F.

[0135] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 N is X 20 is A. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 N is X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 N is X 34 It is F.

[0136] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K and X 34 It is F.

[0137] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 E is X 34 It is F.

[0138] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 A is X 34 It is F.

[0139] In some embodiments, X 15 W is X 16 D is X 20In some embodiments, X 15 W is X 16 D is X 34 It is F.

[0140] In some embodiments, X 15 W is X 16 is K and X 20 In some embodiments, X 15 W is X 16 is K and X 20 In some embodiments, X 15 W is X 16 is K and X 34 It is F.

[0141] In some embodiments, X 15 W is X 16 N is X 20 In some embodiments, X 15 W is X 16 N is X 20 In some embodiments, X 15 W is X 16 N is X 34 It is F.

[0142] In some embodiments, X 15 W is X 20 is K and X 34 It is F.

[0143] In some embodiments, X 15 W is X 20 E is X 34 It is F.

[0144] In some embodiments, X 15 W is X 20 A is X 34 It is F.

[0145] In some embodiments, X 16 is D, X 20 is K and X 34 It is F.

[0146] In some embodiments, X 16 is K and X 20 E is X 34 It is F.

[0147] In some embodiments, X 16 is K and X 20 D is X 34 It is F.

[0148] In some embodiments, X 16 N is X 20 A is X 34 It is F.

[0149] In some embodiments, X 16 N is X 20 is K and X 34 It is F.

[0150] In a particular embodiment, the present disclosure relates to the amino acid sequence of SEQ ID NO: 91: PKPEX5PX7X8DASPX 13 EX 15 X 16 RYYX 20 DX 22 RHYLNWLTRQRX 34 Provides an isolated polypeptide containing -(OH / NH2)(SEQ ID NO: 91), or a pharmaceutically acceptable salt thereof. (In the formula, X5 is either A or K; X7 is either G or K; X8 is E, K, or k; X 13 is either E or K; X 15 is L or W; X 16These are D, E, K, N, S, α-methylserine, or homoserine; X 20 is A, D, E, K, or k; X 22 is A or L; X 34 These are F, 3-pyridinylalanine, 4-pyridinylalanine, 4-carboxyphenylalanine, 4-fluorophenylalanine, 4-methylphenylalanine, N-methylphenylalanine, homophenylalanine, β-homotyrosine, homotyrosine, or N-methyltyrosine; Here, X5, X7, X8, X 13 , or X 20 When K is present, the lysine residue is optionally covalently bonded to the lipophilic substituent via a spacer. Here, X8 or X 20 When k, the D-lysine residue is optionally covalently bonded to the lipophilic substituent via a spacer. However, the polypeptide is provided that it optionally contains at least one residue covalently bonded to a lipophilic substituent via a spacer; The polypeptide optionally further comprises lactam crosslinks formed via amide bonds between the lysine side chain and the aspartic acid side chain, or between the lysine side chain and the glutamic acid side chain, wherein the residues forming the lactam crosslinks are X 16 and X 20 (To be in a certain position).

[0151] In some embodiments, X 15 If X is L, 22 It is A.

[0152] In some embodiments, X 16 If X is N, 20 It is not A.

[0153] In some embodiments, X8 is E, X5 is K, and X 20 If it is K, then X5 or X 20One of the lysine residues is optionally covalently bonded to a lipophilic substituent via a spacer.

[0154] In some embodiments, X8 is K. In some embodiments, the lysine residue at position 8 of the polypeptide sequence is optionally covalently bonded to a lipophilic substituent via a spacer.

[0155] In some embodiments, X 15 It is W.

[0156] In some embodiments, X 34 It is F.

[0157] In some embodiments, the peptide is X 16 and X 20 The peptide further comprises lactam crosslinks formed at the X position via amide bonds between the lysine side chain and the aspartic acid side chain, respectively. In some embodiments, the peptide is X 16 and X 20 The peptide further comprises lactam crosslinks formed at the X position via amide bonds between the lysine side chain and the glutamic acid side chain, respectively. In some embodiments, the peptide is X 16 and X 20 The peptide further comprises lactam crosslinks formed at the X position via amide bonds between the aspartic acid side chain and the lysine side chain, respectively. In some embodiments, the peptide is X 16 and X 20 The material further includes lactam crosslinks formed at the glutamic acid side chain and the lysine side chain via amide bonds.

[0158] In some embodiments, X5 is A. In some embodiments, X5 is K. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, which is optional.

[0159] In some embodiments, X7 is G. In some embodiments, X7 is K. In some embodiments, X7 is K covalently bonded to a lipophilic substituent via a spacer, which is optional.

[0160] In some embodiments, X8 is E. In some embodiments, X8 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer (optional). In some embodiments, X8 is k. In some embodiments, X8 is k covalently bonded to a lipophilic substituent via a spacer (optional).

[0161] In some embodiments, X 13 In some embodiments, X 13 In some embodiments, X 13 This is K, which is optionally covalently bonded to a lipophilic substituent via a spacer.

[0162] In some embodiments, X 15 L is. In some embodiments, X 15 It is W.

[0163] In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 X is α-methylserine. In some embodiments, X 16 It is homoserine.

[0164] In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20In some embodiments, X 20 This is K, which is optionally covalently bonded to a lipophilic substituent via a spacer. In some embodiments, X 20 is k. In some embodiments, X 20 k is optionally covalently bonded to a lipophilic substituent via a spacer.

[0165] In some embodiments, X 22 In some embodiments, X 22 It is L.

[0166] In some embodiments, X 34 In some embodiments, X 34 is 3-pyridinylalanine. In some embodiments, X 34 is 4-pyridinylalanine. In some embodiments, X 34 X is 4-carboxyphenylalanine. In some embodiments, X 34 X is 4-fluorophenylalanine. In some embodiments, X 34 X is 4-methylphenylalanine. In some embodiments, X 34 is N-methylphenylalanine. In some embodiments, X 34 X is homophenylalanine. In some embodiments, X 34 is β-homotyrosine. In some embodiments, X 34 is homotyrosine. In some embodiments, X 34 It is N-methyltyrosine.

[0167] In some embodiments, a carboxy-terminal amino acid, namely X 34 is, -X 34 It is -(NH2). In some embodiments, the carboxy-terminal amino acid X 34 is, -X 34 It is -(OH).

[0168] In some embodiments, certain amino acids represented by the consensus sequence of Sequence ID No. 91 include the following:

[0169] In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, optionally. In some embodiments, X5 is A, and X 15 is W. In some embodiments, X5 is A and X 16 is D. In some embodiments, X5 is A and X 16 is K. In some embodiments, X5 is A and X 16 is N. In some embodiments, X5 is A and X 20 is K. In some embodiments, X5 is A and X 20 is E. In some embodiments, X5 is A and X 20 is A. In some embodiments, X5 is A, and X 34 It is F.

[0170] In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X8 is E. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 is W. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 34 It is F.

[0171] In some embodiments, X5 is K and X8 is E. In some embodiments, X5 is K and X 15 is W. In some embodiments, X5 is K and X 16 is N. In some embodiments, X5 is K and X 16 is D. In some embodiments, X5 is K and X 20 is K. In some embodiments, X5 is K and X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X5 is K and X 34 It is F.

[0172] In some embodiments, X8 is E and X 15 is W. In some embodiments, X8 is E and X 16 is D. In some embodiments, X8 is E and X 16 is N. In some embodiments, X8 is E and X 20 is K. In some embodiments, X8 is E and X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X8 is E and X 34 It is F.

[0173] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 is W. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is E. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is A. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 34 It is F.

[0174] In some embodiments, X 15 W is X 16 In some embodiments, X 15 W is X 16 In some embodiments, X 15 W is X 16 In some embodiments, X 15 W is X 20 In some embodiments, X 15 W is X 20 In some embodiments, X 15 W is X 20 In some embodiments, X 15 W is X 34 It is F.

[0175] In some embodiments, X 16 is D, X 20 In some embodiments, X 16 is D, X 34 It is F.

[0176] In some embodiments, X 16 is K and X 20 In some embodiments, X 16 is K and X 20 In some embodiments, X 16 is K and X 34 It is F.

[0177] In some embodiments, X 16 N is X 20 In some embodiments, X 16 N is X 20 In some embodiments, X 16 N is X 34 It is F.

[0178] In some embodiments, X 20 is K and X 34 It is F.

[0179] In some embodiments, X 20 E is X 34 It is F.

[0180] In some embodiments, X 20 A is X 34 It is F.

[0181] In some embodiments, certain amino acids represented by the consensus sequence of Sequence ID No. 91 include the following: In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, X 15 is W. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20is K. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is E. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is A. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 34 It is F.

[0182] In some embodiments, X5 is A, and X 15 W is X 16 is D. In some embodiments, X5 is A and X 15 W is X 16 is D. In some embodiments, X5 is A and X 15 W is X 16 is K. In some embodiments, X5 is A and X 15 W is X 16 is N. In some embodiments, X5 is A and X 15 W is X 20 is K. In some embodiments, X5 is A and X 15 W is X 20 is E. In some embodiments, X5 is A and X 15 W is X 20 is A. In some embodiments, X5 is A, and X 15 W is X 34 It is F.

[0183] In some embodiments, X5 is A, and X 16 D is X 20 is K. In some embodiments, X5 is A and X 16 D is X 34 It is F.

[0184] In some embodiments, X5 is A, and X 16 is K and X 20 is E. In some embodiments, X5 is A and X 16 is K and X 20 is D. In some embodiments, X5 is A and X 16 is K and X 34 It is F.

[0185] In some embodiments, X5 is A, and X 16 N is X 20 is A. In some embodiments, X5 is A, and X 16 N is X 20 is K. In some embodiments, X5 is A and X 16 N is X 34 It is F.

[0186] In some embodiments, X5 is A, and X 20 is K and X 34 It is F.

[0187] In some embodiments, X5 is A, and X 20 E is X 34 It is F.

[0188] In some embodiments, X5 is A, and X 20 A is X 34 It is F.

[0189] In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 15 is W. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 16 is N. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 16is D. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 20 is K. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 34 It is F.

[0190] In some embodiments, X5 is K, X8 is E, and X 15 is W. In some embodiments, X5 is K, X8 is E, and X 16 is N. In some embodiments, X5 is K, X8 is E, and X 16 is D. In some embodiments, X5 is K, X8 is E, and X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X5 is K, X8 is E, and X 34 It is F.

[0191] In some embodiments, X8 is E and X 15 W is X 16 is N. In some embodiments, X8 is E and X 15 W is X 20 is K. In some embodiments, X8 is E and X 15 W is X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X8 is E and X 15 W is X 34 It is F.

[0192] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 16 is D. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X16 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 16 is N. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 20 is E. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 20 is A. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 34 It is F.

[0193] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D, X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D, X 34 It is F.

[0194] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K, and X 20 is E. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K, and X 20 is D. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K, and X 34 It is F.

[0195] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 N is X 20 is A. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 N is X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 N is X 34 It is F.

[0196] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K and X 34 It is F.

[0197] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 E is X 34 It is F.

[0198] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 A is X 34 It is F.

[0199] In some embodiments, X 15 W is X 16 D is X 20 In some embodiments, X 15 W is X 16 D is X 34 It is F.

[0200] In some embodiments, X 15 W is X 16 is K and X 20In some embodiments, X 15 W is X 16 is K and X 20 In some embodiments, X 15 W is X 16 is K and X 34 It is F.

[0201] In some embodiments, X 15 W is X 16 N is X 20 In some embodiments, X 15 W is X 16 N is X 20 In some embodiments, X 15 W is X 16 N is X 34 It is F.

[0202] In some embodiments, X 15 W is X 20 is K and X 34 It is F.

[0203] In some embodiments, X 15 W is X 20 E is X 34 It is F.

[0204] In some embodiments, X 15 W is X 20 A is X 34 It is F.

[0205] In some embodiments, X 16 is D, X 20 is K and X 34 It is F.

[0206] In some embodiments, X 16 is K and X 20 E is X 34 It is F.

[0207] In some embodiments, X 16 is K and X 20 D is X 34 It is F.

[0208] In some embodiments, X 16 N is X 20 A is X 34 It is F.

[0209] In some embodiments, X 16 N is X 20 is K and X 34 It is F.

[0210] In a particular embodiment, the present disclosure relates to the amino acid sequence of SEQ ID NO: 92: PKPEX5PX7X8DASPX 13 EX 15 X 16 RYYX 20 DX 22 RHYLNWLTRQRX 34 Provides an isolated polypeptide containing -(OH / NH2)(SEQ ID NO: 92), or a pharmaceutically acceptable salt thereof. (In the formula, X5 is either A or K; X7 is either G or K; X8 is E, K, or k; X 13 is either E or K; X 15 is L or W; X 16 These are D, E, K, S, α-methylserine, or homoserine; X 20 is A, D, E, K, or k; X 22 is A or L; X 34These are F, 3-pyridinylalanine, 4-pyridinylalanine, 4-carboxyphenylalanine, 4-fluorophenylalanine, 4-methylphenylalanine, N-methylphenylalanine, homophenylalanine, β-homotyrosine, homotyrosine, or N-methyltyrosine; Here, X5, X7, X8, X 13 , or X 20 When K is present, the lysine residue is optionally covalently bonded to the lipophilic substituent via a spacer. Here, X8 or X 20 When k, the D-lysine residue is optionally covalently bonded to the lipophilic substituent via a spacer. However, the polypeptide is provided that it optionally contains at least one residue covalently bonded to a lipophilic substituent via a spacer; The polypeptide optionally further comprises lactam crosslinks formed via amide bonds between the lysine side chain and the aspartic acid side chain, or between the lysine side chain and the glutamic acid side chain, wherein the residues forming the lactam crosslinks are X 16 and X 20 (To be in a certain position).

[0211] In some embodiments, X 15 If X is L, 22 It is A.

[0212] In some embodiments, X8 is E, X5 is K, and X 20 If it is K, then X5 or X 20 One of the lysine residues is optionally covalently bonded to a lipophilic substituent via a spacer.

[0213] In some embodiments, X8 is K. In some embodiments, the lysine residue at position 8 of the polypeptide sequence is optionally covalently bonded to a lipophilic substituent via a spacer.

[0214] In some embodiments, X 15 It is W.

[0215] In some embodiments, X 34 It is F.

[0216] In some embodiments, the peptide is X 16 and X 20 The peptide further comprises lactam crosslinks formed at the X position via amide bonds between the lysine side chain and the aspartic acid side chain, respectively. In some embodiments, the peptide is X 16 and X 20 The peptide further comprises lactam crosslinks formed at the X position via amide bonds between the lysine side chain and the glutamic acid side chain, respectively. In some embodiments, the peptide is X 16 and X 20 The peptide further comprises lactam crosslinks formed at the X position via amide bonds between the aspartic acid side chain and the lysine side chain, respectively. In some embodiments, the peptide is X 16 and X 20 The material further includes lactam crosslinks formed at the glutamic acid side chain and the lysine side chain via amide bonds.

[0217] In some embodiments, X5 is A. In some embodiments, X5 is K. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, which is optional.

[0218] In some embodiments, X7 is G. In some embodiments, X7 is K. In some embodiments, X7 is K covalently bonded to a lipophilic substituent via a spacer, which is optional.

[0219] In some embodiments, X8 is E. In some embodiments, X8 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer (optional). In some embodiments, X8 is k. In some embodiments, X8 is k covalently bonded to a lipophilic substituent via a spacer (optional).

[0220] In some embodiments, X 13 In some embodiments, X 13 In some embodiments, X 13 This is K, which is optionally covalently bonded to a lipophilic substituent via a spacer.

[0221] In some embodiments, X 15 L is. In some embodiments, X 15 It is W.

[0222] In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 X is α-methylserine. In some embodiments, X 16 It is homoserine.

[0223] In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 This is K, which is optionally covalently bonded to a lipophilic substituent via a spacer. In some embodiments, X 20 is k. In some embodiments, X 20 k is optionally covalently bonded to a lipophilic substituent via a spacer.

[0224] In some embodiments, X 22 In some embodiments, X 22 It is L.

[0225] In some embodiments, X 34 In some embodiments, X34 is 3-pyridinylalanine. In some embodiments, X 34 is 4-pyridinylalanine. In some embodiments, X 34 X is 4-carboxyphenylalanine. In some embodiments, X 34 X is 4-fluorophenylalanine. In some embodiments, X 34 X is 4-methylphenylalanine. In some embodiments, X 34 is N-methylphenylalanine. In some embodiments, X 34 X is homophenylalanine. In some embodiments, X 34 is β-homotyrosine. In some embodiments, X 34 is homotyrosine. In some embodiments, X 34 It is N-methyltyrosine.

[0226] In some embodiments, a carboxy-terminal amino acid, namely X 34 is, -X 34 It is -(NH2). In some embodiments, the carboxy-terminal amino acid X 34 is, -X 34 It is -(OH).

[0227] In some embodiments, certain amino acids represented by the consensus sequence of Sequence ID No. 92 include the following:

[0228] In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, optionally. In some embodiments, X5 is A, and X 15 is W. In some embodiments, X5 is A and X 16 is D. In some embodiments, X5 is A and X 16 is K. In some embodiments, X5 is A and X 20 is K. In some embodiments, X5 is A and X 20is E. In some embodiments, X5 is A and X 20 is A. In some embodiments, X5 is A, and X 34 It is F.

[0229] In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X8 is E. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 is W. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 34 It is F.

[0230] In some embodiments, X5 is K and X8 is E. In some embodiments, X5 is K and X 15 is W. In some embodiments, X5 is K and X 16 is D. In some embodiments, X5 is K and X 20 is K. In some embodiments, X5 is K and X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X5 is K and X 34 It is F.

[0231] In some embodiments, X8 is E and X 15 is W. In some embodiments, X8 is E and X 16 is D. In some embodiments, X8 is E and X 20 is K. In some embodiments, X8 is E and X 20is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X8 is E and X 34 It is F.

[0232] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 is W. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is E. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is A. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 34 It is F.

[0233] In some embodiments, X 15 W is X 16 In some embodiments, X 15 W is X 16 In some embodiments, X 15 W is X 20 In some embodiments, X 15 W is X 20 In some embodiments, X 15 W is X 20 In some embodiments, X 15 W is X 34 It is F.

[0234] In some embodiments, X 16 is D, X 20 In some embodiments, X 16 is D, X 34 It is F.

[0235] In some embodiments, X 16 is K and X 20 In some embodiments, X 16 is K and X 20 In some embodiments, X 16 is K and X 34 It is F.

[0236] In some embodiments, X 20 is K and X 34 It is F.

[0237] In some embodiments, X 20 E is X 34 It is F.

[0238] In some embodiments, X 20 A is X 34 It is F.

[0239] In some embodiments, certain amino acids represented by the consensus sequence of Sequence ID No. 92 include the following: In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, X 15 is W. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20is K. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is E. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is A. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 34 It is F.

[0240] In some embodiments, X5 is A, and X 15 W is X 16 is D. In some embodiments, X5 is A and X 15 W is X 16 is D. In some embodiments, X5 is A and X 15 W is X 16 is K. In some embodiments, X5 is A and X 15 W is X 20 is K. In some embodiments, X5 is A and X 15 W is X 20 is E. In some embodiments, X5 is A and X 15 W is X 20 is A. In some embodiments, X5 is A, and X 15 W is X 34 It is F.

[0241] In some embodiments, X5 is A, and X 16 D is X 20 is K. In some embodiments, X5 is A and X 16 D is X 34 It is F.

[0242] In some embodiments, X5 is A, and X 16 is K and X 20is E. In some embodiments, X5 is A and X 16 is K and X 20 is D. In some embodiments, X5 is A and X 16 is K and X 34 It is F.

[0243] In some embodiments, X5 is A, and X 20 is K and X 34 It is F.

[0244] In some embodiments, X5 is A, and X 20 E is X 34 It is F.

[0245] In some embodiments, X5 is A, and X 20 A is X 34 It is F.

[0246] In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 15 is W. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 16 is D. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 20 is K. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 34 It is F.

[0247] In some embodiments, X5 is K, X8 is E, and X 15 is W. In some embodiments, X5 is K, X8 is E, and X 16 is D. In some embodiments, X5 is K, X8 is E, and X 20is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X5 is K, X8 is E, and X 34 It is F.

[0248] In some embodiments, X8 is E and X 15 W is X 20 is K. In some embodiments, X8 is E and X 15 W is X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X8 is E and X 15 W is X 34 It is F.

[0249] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 16 is D. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 16 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 20 is E. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 20 is A. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 34 It is F.

[0250] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D, X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D, X 34 It is F.

[0251] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K, and X 20 is E. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K, and X 20 is D. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K, and X 34 It is F.

[0252] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K and X 34 It is F.

[0253] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 E is X 34 It is F.

[0254] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 A is X 34 It is F.

[0255] In some embodiments, X 15 W is X 16 D is X 20 In some embodiments, X15 W is X 16 D is X 34 It is F.

[0256] In some embodiments, X 15 W is X 16 is K and X 20 In some embodiments, X 15 W is X 16 is K and X 20 In some embodiments, X 15 W is X 16 is K and X 34 It is F.

[0257] In some embodiments, X 15 W is X 20 is K and X 34 It is F.

[0258] In some embodiments, X 15 W is X 20 E is X 34 It is F.

[0259] In some embodiments, X 15 W is X 20 A is X 34 It is F.

[0260] In some embodiments, X 16 is D, X 20 is K and X 34 It is F.

[0261] In some embodiments, X 16 is K and X 20 E is X 34 It is F.

[0262] In some embodiments, X 16 is K and X 20 D is X 34is F.

[0263] In a specific embodiment, the present disclosure provides an isolated polypeptide comprising the amino acid sequence of SEQ ID NO: 93: PKPEX5PX7X8DASPX 13 EX 15 X 16 RYYX 20 DX 22 RHYLNWLTRQRX 34 -(OH / NH2) (SEQ ID NO: 93), or a pharmaceutically acceptable salt thereof wherein X5 is A or K; X7 is G or K; X8 is E, K, or k; X 13 is E or K; X 15 is L or W; X 16 is D, E, K, N, S, α-methylserine, or homoserine; X 20 is D, E, K, or k; X 22 is A or L; X 34 is F, 3-pyridinylalanine, 4-pyridinylalanine, 4-carboxyphenylalanine, 4-fluorophenylalanine, 4-methylphenylalanine, N-methylphenylalanine, homophenylalanine, β-homotyrosine, homotyrosine, or N-methyltyrosine; wherein, when X5, X7, X8, X 13 , or X 20 is K, the lysine residue is optionally covalently bonded to a lipophilic substituent, optionally via a spacer, wherein, when X8 or X 20 is k, the D-lysine residue is optionally covalently bonded to a lipophilic substituent, optionally via a spacer, provided that the polypeptide comprises at least one residue covalently bonded to a lipophilic substituent, optionally via a spacer; The polypeptide optionally further comprises lactam crosslinks formed via amide bonds between the lysine side chain and the aspartic acid side chain, or between the lysine side chain and the glutamic acid side chain, wherein the residues forming the lactam crosslinks are X 16 and X 20 (To be in a certain position).

[0264] In some embodiments, X 15 If X is L, 22 It is A.

[0265] In some embodiments, X8 is E, X5 is K, and X 20 If it is K, then X5 or X 20 One of the lysine residues is optionally covalently bonded to a lipophilic substituent via a spacer.

[0266] In some embodiments, X8 is K. In some embodiments, the lysine residue at position 8 of the polypeptide sequence is optionally covalently bonded to a lipophilic substituent via a spacer.

[0267] In some embodiments, X 15 It is W.

[0268] In some embodiments, X 34 It is F.

[0269] In some embodiments, the peptide is X 16 and X 20 The peptide further comprises lactam crosslinks formed at the X position via amide bonds between the lysine side chain and the aspartic acid side chain, respectively. In some embodiments, the peptide is X 16 and X 20 The peptide further comprises lactam crosslinks formed at the X position via amide bonds between the lysine side chain and the glutamic acid side chain, respectively. In some embodiments, the peptide is X 16 and X 20The peptide further comprises lactam crosslinks formed at the X position via amide bonds between the aspartic acid side chain and the lysine side chain, respectively. In some embodiments, the peptide is X 16 and X 20 The material further includes lactam crosslinks formed at the glutamic acid side chain and the lysine side chain via amide bonds.

[0270] In some embodiments, X5 is A. In some embodiments, X5 is K. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, which is optional.

[0271] In some embodiments, X7 is G. In some embodiments, X7 is K. In some embodiments, X7 is K covalently bonded to a lipophilic substituent via a spacer, which is optional.

[0272] In some embodiments, X8 is E. In some embodiments, X8 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer (optional). In some embodiments, X8 is k. In some embodiments, X8 is k covalently bonded to a lipophilic substituent via a spacer (optional).

[0273] In some embodiments, X 13 In some embodiments, X 13 In some embodiments, X 13 This is K, which is optionally covalently bonded to a lipophilic substituent via a spacer.

[0274] In some embodiments, X 15 L is. In some embodiments, X 15 It is W.

[0275] In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X16 In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 X is α-methylserine. In some embodiments, X 16 It is homoserine.

[0276] In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 This is K, optionally covalently bonded to a lipophilic substituent via a spacer. In some embodiments, X 20 is k. In some embodiments, X 20 k is optionally covalently bonded to a lipophilic substituent via a spacer.

[0277] In some embodiments, X 22 In some embodiments, X 22 It is L.

[0278] In some embodiments, X 34 In some embodiments, X 34 is 3-pyridinylalanine. In some embodiments, X 34 is 4-pyridinylalanine. In some embodiments, X 34 X is 4-carboxyphenylalanine. In some embodiments, X 34 X is 4-fluorophenylalanine. In some embodiments, X 34 X is 4-methylphenylalanine. In some embodiments, X 34 is N-methylphenylalanine. In some embodiments, X 34 X is homophenylalanine. In some embodiments, X 34 is β-homotyrosine. In some embodiments, X34 is homothyrosine. In some embodiments, X 34 is N-methyltyrosine.

[0279] In some embodiments, the carboxy-terminal amino acid, i.e., X 34 is -X 34 -(NH2). In some embodiments, the carboxy-terminal amino acid X 34 is -X 34 -(OH).

[0280] In some embodiments, certain amino acids represented by the consensus sequence of SEQ ID NO: 93 include: In some embodiments, X5 is A, and X8 is K covalently linked to a lipophilic substituent, optionally via a spacer. In some embodiments, X5 is A, and X 15 is W. In some embodiments, X5 is A, and X 16 is D. In some embodiments, X5 is A, and X 16 is K. In some embodiments, X5 is A, and X 16 is N. In some embodiments, X5 is A, and X 20 is K. In some embodiments, X5 is A, and X 20 is E. X5 is A, and X 34 is F.

[0281] In some embodiments, X5 is K covalently linked to a lipophilic substituent optionally via a spacer, and X8 is E. In some embodiments, X5 is K covalently linked to a lipophilic substituent optionally via a spacer, and X 15 is W. In some embodiments, X5 is K covalently linked to a lipophilic substituent optionally via a spacer, and X 16 is N. In some embodiments, X5 is K covalently linked to a lipophilic substituent optionally via a spacer, and X 16is D. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 34 It is F.

[0282] In some embodiments, X5 is K and X8 is E. In some embodiments, X5 is K and X 15 is W. In some embodiments, X5 is K and X 16 is N. In some embodiments, X5 is K and X 16 is D. In some embodiments, X5 is K and X 20 is K. In some embodiments, X5 is K and X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X5 is K and X 34 It is F.

[0283] In some embodiments, X8 is E and X 15 is W. In some embodiments, X8 is E and X 16 is D. In some embodiments, X8 is E and X 16 is N. In some embodiments, X8 is E and X 20 is K. In some embodiments, X8 is E and X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X8 is E and X 34 It is F.

[0284] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 is W. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16is D. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is E. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 34 It is F.

[0285] In some embodiments, X 15 W is X 16 In some embodiments, X 15 W is X 16 In some embodiments, X 15 W is X 16 In some embodiments, X 15 W is X 20 In some embodiments, X 15 W is X 20 In some embodiments, X 15 W is X 34 It is F.

[0286] In some embodiments, X 16 is D, X 20 In some embodiments, X 16 is D, X 34 It is F.

[0287] In some embodiments, X 16 is K and X 20 In some embodiments, X 16 is K and X 20In some embodiments, X 16 is K and X 34 It is F.

[0288] In some embodiments, X 16 is N and X 20 In some embodiments, X 16 is N and X 34 It is F.

[0289] In some embodiments, X 20 is K and X 34 It is F.

[0290] In some embodiments, X 20 E is X 34 It is F.

[0291] In some embodiments, certain amino acids represented by the consensus sequence of Sequence ID No. 93 include the following:

[0292] In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, X 15 is W. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20is E. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 34 It is F.

[0293] In some embodiments, X5 is A, and X 15 W is X 16 is D. In some embodiments, X5 is A and X 15 W is X 16 is D. In some embodiments, X5 is A and X 15 W is X 16 is K. In some embodiments, X5 is A and X 15 W is X 16 is N. In some embodiments, X5 is A and X 15 W is X 20 is K. In some embodiments, X5 is A and X 15 W is X 20 is E. In some embodiments, X5 is A and X 15 W is X 34 It is F.

[0294] In some embodiments, X5 is A, and X 16 D is X 20 is K. In some embodiments, X5 is A and X 16 D is X 34 It is F.

[0295] In some embodiments, X5 is A, and X 16 is K and X 20 is E. In some embodiments, X5 is A and X 16 is K and X 20 is D. In some embodiments, X5 is A and X 16 is K and X 34 It is F.

[0296] In some embodiments, X5 is A, and X 16 is N and X 20 is K. In some embodiments, X5 is A and X 16 is N and X 34 It is F.

[0297] In some embodiments, X5 is A, and X 20 is K and X 34 It is F.

[0298] In some embodiments, X5 is A, and X 20 E is X 34 It is F.

[0299] In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 15 is W. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 16 is N. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 16 is D. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 20 is K. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 34 It is F.

[0300] In some embodiments, X5 is K, X8 is E, and X 15 is W. In some embodiments, X5 is K, X8 is E, and X 16 is N. In some embodiments, X5 is K, X8 is E, and X 16 is D. In some embodiments, X5 is K, X8 is E, and X 20is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X5 is K, X8 is E, and X 34 It is F.

[0301] In some embodiments, X8 is E and X 15 W is X 16 is N. In some embodiments, X8 is E and X 15 W is X 20 is K. In some embodiments, X8 is E and X 15 W is X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X8 is E and X 15 W is X 34 It is F.

[0302] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 16 is D. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 16 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 16 is N. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 20 is E. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 15 W is X 34 It is F.

[0303] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D, X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D, X 34 It is F.

[0304] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K, and X 20 is E. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K, and X 20 is D. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K, and X 34 It is F.

[0305] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N and X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N and X 34 It is F.

[0306] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K and X 34 It is F.

[0307] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 E is X 34 It is F.

[0308] In some embodiments, X 15 W is X 16 D is X 20 In some embodiments, X 15 W is X 16 D is X 34 It is F.

[0309] In some embodiments, X 15 W is X 16 is K and X 20 In some embodiments, X 15 W is X 16 is K and X 20 In some embodiments, X 15 W is X 16 is K and X 34 It is F.

[0310] In some embodiments, X 15 W is X 16 N is X 20 In some embodiments, X 15 W is X 16 N is X 34 It is F.

[0311] In some embodiments, X 15 W is X 20 is K and X 34 It is F.

[0312] In some embodiments, X 15 W is X 20 E is X 34 It is F.

[0313] In some embodiments, X 16 is D, X 20 is K and X 34 It is F.

[0314] In some embodiments, X 16 is K and X 20 E is X 34 It is F.

[0315] In some embodiments, X 16 is K and X 20 D is X 34 It is F.

[0316] In some embodiments, X 16 N is X 20 is K and X 34 It is F.

[0317] In a particular embodiment, the present invention relates to the amino acid sequence of SEQ ID NO: 94: PKPEX5PGX8DASPX 13 EW Test 16 RYYX 20 DX 22 RHYLNWLTRQRX 34 Provides an isolated polypeptide containing -(OH / NH2)(SEQ ID NO: 94), or a pharmaceutically acceptable salt thereof. (In the formula, X5 is either A or K; X8 is E, K, or k; X 13 is either E or K; X 16 is D, E, K, or N; X 20 is A, D, E, K, or k; X 22 is A or L; X 34 is F or N-methyltyrosine; Here, X5, X8, X 13 , or X 20 When K is present, the lysine residue is optionally covalently bonded to the lipophilic substituent via a spacer. Here, X8 or X 20When k, the D-lysine residue is optionally covalently bonded to the lipophilic substituent via a spacer. However, the polypeptide is provided that it optionally contains at least one residue covalently bonded to a lipophilic substituent via a spacer; The polypeptide optionally further comprises lactam crosslinks formed via amide bonds between the lysine side chain and the aspartic acid side chain, or between the lysine side chain and the glutamic acid side chain, wherein the residues forming the lactam crosslinks are X 16 and X 20 (To be in a certain position).

[0318] In some embodiments, X 16 If X is N, 20 It is not A.

[0319] In some embodiments, X8 is E, X5 is K, and X 20 If it is K, then X5 or X 20 One of the lysine residues is optionally covalently bonded to a lipophilic substituent via a spacer.

[0320] In some embodiments, X8 is K. In some embodiments, the lysine residue at position 8 of the polypeptide sequence is optionally covalently bonded to a lipophilic substituent via a spacer.

[0321] In some embodiments, X 34 It is F.

[0322] In some embodiments, the peptide is X 16 and X 20 The peptide further comprises lactam crosslinks formed at the X position via amide bonds between the lysine side chain and the aspartic acid side chain, respectively. In some embodiments, the peptide is X 16 and X 20 The peptide further comprises lactam crosslinks formed at the X position via amide bonds between the lysine side chain and the glutamic acid side chain, respectively. In some embodiments, the peptide is X 16 and X 20The peptide further comprises lactam crosslinks formed at the X position via amide bonds between the aspartic acid side chain and the lysine side chain, respectively. In some embodiments, the peptide is X 16 and X 20 The material further includes lactam crosslinks formed at the glutamic acid side chain and the lysine side chain via amide bonds.

[0323] In some embodiments, X5 is A. In some embodiments, X5 is K. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, which is optional.

[0324] In some embodiments, X8 is E. In some embodiments, X8 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer (optional). In some embodiments, X8 is k. In some embodiments, X8 is k covalently bonded to a lipophilic substituent via a spacer (optional).

[0325] In some embodiments, X 13 In some embodiments, X 13 In some embodiments, X 13 This is K, which is optionally covalently bonded to a lipophilic substituent via a spacer.

[0326] In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 It is N.

[0327] In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X20 This is K, which is optionally covalently bonded to a lipophilic substituent via a spacer. In some embodiments, X 20 is k. In some embodiments, X 20 k is optionally covalently bonded to a lipophilic substituent via a spacer.

[0328] In some embodiments, X 22 In some embodiments, X 22 It is L.

[0329] In some embodiments, X 34 In some embodiments, X 34 It is N-methyltyrosine.

[0330] In some embodiments, a carboxy-terminal amino acid, namely X 34 is, -X 34 It is -(NH2). In some embodiments, the carboxy-terminal amino acid X 34 is, -X 34 It is -(OH).

[0331] In some embodiments, certain amino acids represented by the consensus sequence of Sequence ID No. 94 include the following: In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, optionally. In some embodiments, X5 is A, and X 16 is D. In some embodiments, X5 is A and X 16 is K. In some embodiments, X5 is A and X 16 is N. In some embodiments, X5 is A and X 20 is K. In some embodiments, X5 is A and X 20 is E. In some embodiments, X5 is A and X 20 is A. In some embodiments, X5 is A, and X 34 It is F.

[0332] In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X8 is E. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 34 It is F.

[0333] In some embodiments, X5 is K and X8 is E. In some embodiments, X5 is K and X 16 is N. In some embodiments, X5 is K and X 16 is D. In some embodiments, X5 is K and X 20 is K. In some embodiments, X5 is K and X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X5 is K and X 34 It is F.

[0334] In some embodiments, X8 is E and X 16 is D. In some embodiments, X8 is E and X 16 is N. In some embodiments, X8 is E and X 20 is K. In some embodiments, X8 is E and X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X8 is E and X 34 It is F.

[0335] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is E. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is A. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 34 It is F.

[0336] In some embodiments, X 16 D is X 20 In some embodiments, X 16 D is X 34 It is F.

[0337] In some embodiments, X 16 is K and X 20 In some embodiments, X 16 is K and X 20 In some embodiments, X 16 is K and X 34 It is F.

[0338] In some embodiments, X 16 N is X 20 In some embodiments, X 16 N is X 20 In some embodiments, X16 N is X 34 It is F.

[0339] In some embodiments, X 20 is K and X 34 It is F.

[0340] In some embodiments, X 20 E is X 34 It is F.

[0341] In some embodiments, X 20 A is X 34 It is F.

[0342] In some embodiments, certain amino acids represented by the consensus sequence of Sequence ID No. 94 include the following:

[0343] In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, X 16 is D. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is E. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is A. In some embodiments, X5 is A, and X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 34 It is F.

[0344] In some embodiments, X5 is A, and X 16 D is X 20 is K. In some embodiments, X5 is A and X 16 D is X 34 It is F.

[0345] In some embodiments, X5 is A, and X 16 is K and X 20 is E. In some embodiments, X5 is A and X 16 is K and X 20 is D. In some embodiments, X5 is A and X 16 is K and X 34 It is F.

[0346] In some embodiments, X5 is A, and X 16 N is X 20 is A. In some embodiments, X5 is A, and X 16 N is X 20 is K. In some embodiments, X5 is A and X 16 N is X 34 It is F.

[0347] In some embodiments, X5 is A, and X 20 is K and X 34 It is F.

[0348] In some embodiments, X5 is A, and X 20 E is X 34 It is F.

[0349] In some embodiments, X5 is A, and X 20 A is X 34 It is F.

[0350] In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 16 is N. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 16 is D. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 20 is K. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, X8 is E, and X 34 It is F.

[0351] In some embodiments, X5 is K, X8 is E, and X 16 is N. In some embodiments, X5 is K, X8 is E, and X 16 is D. In some embodiments, X5 is K, X8 is E, and X 20 is K covalently bonded to a lipophilic substituent via a spacer, which is optional. In some embodiments, X5 is K, X8 is E, and X 34 It is F.

[0352] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D, X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D, X 34 It is F.

[0353] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K, and X 20 is E. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K, and X20 is D. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is K, and X 34 It is F.

[0354] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N, and X 20 is A. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N, and X 20 is K. In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N, and X 34 It is F.

[0355] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 is K and X 34 It is F.

[0356] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 E is X 34 It is F.

[0357] In some embodiments, X8 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 A is X 34 It is F.

[0358] In some embodiments, X 16 is D, X 20 is K and X 34 It is F.

[0359] In some embodiments, X 16 is K and X 20E is X 34 It is F.

[0360] In some embodiments, X 16 is K and X 20 D is X 34 It is F.

[0361] In some embodiments, X 16 N is X 20 A is X 34 It is F.

[0362] In some embodiments, X 16 N is X 20 is K and X 34 It is F.

[0363] In a particular embodiment, the present disclosure relates to the amino acid sequence of SEQ ID NO: 95: PKPEX5PGK8DASPX 13 EW Test 16 RYYX 20 This provides an isolated polypeptide containing DLRHYLNWLTRQRF-(OH / NH2)(SEQ ID NO: 95), or a pharmaceutically acceptable salt thereof (wherein, X5 is either A or K; X 13 is either E or K; X 16 is D, E, K, or N; X 20 is A, D, E, K, or k; Here, X5, X 13 or X 20 When K is present, the lysine residue is optionally covalently bonded to the lipophilic substituent via a spacer. Here, K8 is optionally covalently bonded to a lipophilic substituent via a spacer. Here, X 20 When k, the D-lysine residue is optionally covalently bonded to the lipophilic substituent via a spacer. However, the polypeptide is provided that it optionally contains at least one residue covalently bonded to a lipophilic substituent via a spacer; The polypeptide optionally further comprises lactam crosslinks formed via amide bonds between the lysine side chain and the aspartic acid side chain, or between the lysine side chain and the glutamic acid side chain, wherein the residues forming the lactam crosslinks are X 16 and X 20 (To be in a certain position).

[0364] In some embodiments, X 16 If X is N, 20 It is not A.

[0365] In some embodiments, K is optionally covalently bonded to a lipophilic substituent via a spacer.

[0366] In some embodiments, the peptide is X 16 and X 20 The peptide further comprises lactam crosslinks formed at the X position via amide bonds between the lysine side chain and the aspartic acid side chain, respectively. In some embodiments, the peptide is X 16 and X 20 The peptide further comprises lactam crosslinks formed at the X position via amide bonds between the lysine side chain and the glutamic acid side chain, respectively. In some embodiments, the peptide is X 16 and X 20 The peptide further comprises lactam crosslinks formed at the X position via amide bonds between the aspartic acid side chain and the lysine side chain, respectively. In some embodiments, the peptide is X 16 and X 20 The material further includes lactam crosslinks formed at the glutamic acid side chain and the lysine side chain via amide bonds.

[0367] In some embodiments, X5 is A. In some embodiments, X5 is K. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, which is optional.

[0368] In some embodiments, K8 is unsubstituted. In some embodiments, K8 is optionally covalently bonded to a lipophilic substituent via a spacer.

[0369] In some embodiments, X 13 In some embodiments, X 13 In some embodiments, X 13 This is K, which is optionally covalently bonded to a lipophilic substituent via a spacer.

[0370] In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 It is N.

[0371] In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 This is K, which is optionally covalently bonded to a lipophilic substituent via a spacer. In some embodiments, X 20 is k. In some embodiments, X 20 k is optionally covalently bonded to a lipophilic substituent via a spacer.

[0372] In some embodiments, a carboxy-terminal amino acid, namely F 34 is -F 34 It is -(NH2). In some embodiments, the carboxy-terminal amino acid F 34 is -F 34 It is -(OH).

[0373] In some embodiments, certain amino acids represented by the consensus sequence of Sequence ID No. 95 include the following: In some embodiments, X5 is A, and K8 is optionally covalently bonded to a lipophilic substituent via a spacer. In some embodiments, X5 is A, and X 16 is D. In some embodiments, X5 is A and X 16 is K. In some embodiments, X5 is A and X 16 is N. In some embodiments, X5 is A and X 20 is K. In some embodiments, X5 is A and X 20 is E. In some embodiments, X5 is A and X 20 A is the answer.

[0374] In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is N. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 16 is D. In some embodiments, X5 is K covalently bonded to a lipophilic substituent via a spacer, and X 20 K is K.

[0375] In some embodiments, X5 is K, and X 16 is N. In some embodiments, X5 is K and X 16 is D. In some embodiments, X5 is K and X 20 is K. In some embodiments, X5 is K and X 20 This is K, which is optionally covalently bonded to a lipophilic substituent via a spacer.

[0376] In some embodiments, K8 is optionally covalently bonded to a lipophilic substituent via a spacer, X 16 is D. In some embodiments, K8 is optionally covalently bonded to a lipophilic substituent via a spacer, X 16is K. In some embodiments, K8 is optionally covalently bonded to a lipophilic substituent via a spacer, and X 16 is N. In some embodiments, K8 is optionally covalently bonded to a lipophilic substituent via a spacer, and X 20 is K. In some embodiments, K8 is optionally covalently bonded to a lipophilic substituent via a spacer, and X 20 is E. In some embodiments, K8 is optionally covalently bonded to a lipophilic substituent via a spacer, X 20 A is the answer.

[0377] In some embodiments, X 16 is D, X 20 K is K.

[0378] In some embodiments, X 16 is K and X 20 In some embodiments, X 16 is K and X 20 It is D.

[0379] In some embodiments, X 16 N is X 20 In some embodiments, X 16 N is X 20 K is K.

[0380] In some embodiments, certain amino acids represented by the consensus sequence of Sequence ID No. 95 include the following: In some embodiments, X5 is A, and K8 is optionally covalently bonded to a lipophilic substituent via a spacer, X 16 is D. In some embodiments, X5 is A, and K8 is optionally covalently bonded to a lipophilic substituent via a spacer, X 16 is K. In some embodiments, X5 is A, and K8 is optionally covalently bonded to a lipophilic substituent via a spacer, X 16is N. In some embodiments, X5 is A, and K8 is optionally covalently bonded to a lipophilic substituent via a spacer, X 20 is K. In some embodiments, X5 is A, and K8 is optionally covalently bonded to a lipophilic substituent via a spacer, X 20 is E. In some embodiments, X5 is A, and K8 is optionally covalently bonded to a lipophilic substituent via a spacer, X 20 A is the answer.

[0381] In some embodiments, X5 is A, and X 16 D is X 20 K is K.

[0382] In some embodiments, X5 is A, and X 16 is K and X 20 is E. In some embodiments, X5 is A and X 16 is K and X 20 It is D.

[0383] In some embodiments, X5 is A, and X 16 N is X 20 is A. In some embodiments, X5 is A, and X 16 N is X 20 K is K.

[0384] In some embodiments, K8 is optionally covalently bonded to a lipophilic substituent via a spacer, X 16 is D, X 20 K is K.

[0385] In some embodiments, K8 is optionally covalently bonded to a lipophilic substituent via a spacer, X 16 is K, and X 20 is E. In some embodiments, K8 is optionally covalently bonded to a lipophilic substituent via a spacer, X 16 is K, and X 20It is D.

[0386] In some embodiments, K8 is optionally covalently bonded to a lipophilic substituent via a spacer, X 16 N is X 20 A is. In some embodiments, K8 is optionally covalently bonded to a lipophilic substituent via a spacer, and X 16 N is X 20 K is K.

[0387] Optional conjugation of lipophilic substituents to any peptide via a spacer. In some embodiments, any of the disclosed polypeptides may be optionally replaced with one or more lipophilic substituents via spacers, where “lipophilic substituent” and “spacer” are defined herein. In some embodiments, any of the disclosed polypeptides comprising an amino acid sequence selected from the group consisting of amino acid sequences represented by any of the consensus sequences from SEQ ID NO: 1 to SEQ ID NO: 78 may optionally contain or be modified with one or more lipophilic substituents via spacers, or may be further modified by covalent bonding of one or more lipophilic substituents via spacers. In some embodiments, any of the disclosed polypeptides comprising an amino acid sequence selected from the group consisting of amino acid sequences represented by any of the consensus sequences from SEQ ID NO: 101 to SEQ ID NO: 181 may be further modified by covalent bonding of one or more lipophilic substituents via spacers. In some embodiments, any of the disclosed polypeptides, comprising an amino acid sequence selected from the group consisting of amino acid sequences represented by any of the consensus sequences of SEQ ID NOs: 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 280, or 281, can each be optionally modified by covalent bonding of one or more lipophilic substituents via spacers. In some embodiments, the lipophilic substituents can be bonded to an amino group of the polypeptide (e.g., the ε-amino group of a lysine residue) by the carboxyl group of the lipophilic substituent or optionally by the amino group of the spacer, where the carboxyl group of the spacer forms an amide bond with the ε-amino group of the lysine residue.

[0388] lipophilic substituent The conjugation of one or more “lipophilic substituents” to any of the disclosed polypeptides of the present invention, each optionally via a “spacer,” is intended to extend the action of the polypeptide by promoting binding to serum albumin and delaying the renal clearance of the conjugated polypeptide. As used herein, “lipophilic substituents” include substituents containing 4 to 40 carbon atoms, 8 to 25 carbon atoms, 12 to 22 carbon atoms, or 6 to 20 carbon atoms. The lipophilic substituents can be bonded by the carboxyl group of the lipophilic substituent to an amino group of the polypeptide (e.g., the ε-amino group of a lysine residue), or optionally to an amino group of a spacer, the carboxyl group of the spacer further forming an amide bond with the amino group of the amino acid (e.g., lysine) residue to which it is bonded. In some embodiments, the polypeptide comprises three, two, or preferably one lipophilic substituent, each with or without an optional spacer, which are defined in more detail below.

[0389] In some embodiments, the lipophilic substituent includes a linear or branched alkyl group. In some embodiments, the lipophilic substituent is an acyl group of a linear or branched fatty acid. In some embodiments, the lipophilic substituent is an acyl group of a linear or branched fatty acid, further substituted with one or more carboxylic acid and / or hydroxamic acid groups.

[0390] In some embodiments, the polypeptide comprises three, two, or preferably one lipophilic substituent without an optional spacer. In some embodiments, the lipophilic substituent is -CO(CH2) 16 It is CO2H. In some embodiments, the lipophilic substituent is -CO(CH2) 18 It is CO2H. In some embodiments, the lipophilic substituent is -CO(CH2) 20 It is CO2H.

[0391] In some embodiments, the polypeptide comprises three, two, or preferably one lipophilic substituent without an optional spacer. In some embodiments, the lipophilic substituent is of formula I: -CO-(CH2) m -Z Equation I It is a monovalent group. (In the formula, Z is either -CH3 or -CO2H; m is between 4 and 24. This lipophilic substituent forms an amide bond between the amino group of the disclosed polypeptide (e.g., the ε-amino group of lysine) and the CO-group of the lipophilic substituent. In some embodiments, Z is -CO2H. In some embodiments, m is 14-20. In some embodiments, the lipophilic substituent is covalently bonded to the isolated polypeptide via a spacer. In some embodiments, the lipophilic substituent is -CO-(CH2) m -Z is linked to the amino group of the isolated polypeptide via a spacer, and the spacer forms a crosslink between the amino group of the isolated polypeptide and the CO- group of the lipophilic substituent.

[0392] In some embodiments, m is selected from the group consisting of 4-20, 8-20, 12-20, 14-20, 16-20, 14, 16, 18, and 20.

[0393] In some embodiments, Z is -CO2H, and the lipophilic substituent is of the formula -CO-(CH2) m It contains -CO2H. In some embodiments, -CO-(CH2) m -Z is -CO-(CH2)4-CO2H, -CO-(CH2)5-CO2H, -CO-(CH2)6-CO2H, -CO-(CH2)7-CO2H, -CO-(CH2)8-CO2H, -CO-(CH2)9-CO2H, -CO-(CH2) 10 -CO2H, -CO-(CH2) 11 -CO2H, -CO-(CH2) 12 -CO2H, -CO-(CH2) 13-CO2H, -CO-(CH2) 14 -CO2H, -CO-(CH2) 15 -CO2H, -CO-(CH2) 16 -CO2H, -CO-(CH2) 17 -CO2H, -CO-(CH2) 18 -CO2H, -CO-(CH2) 19 -CO2H, -CO-(CH2) 20 Selected from the group consisting of -CO2H.

[0394] In some embodiments, the lipophilic substituent is -CO-(CH2) 14 -CO2H. In some embodiments, the lipophilic substituent is -CO-(CH2) 16 -CO2H. In some embodiments, the lipophilic substituent is -CO-(CH2) 18 -CO2H. In some embodiments, the lipophilic substituent is -CO-(CH2) 20 -CO2H

[0395] In some embodiments, Z is -CH3, and the lipophilic substituent is of the formula -CO-(CH2) m It has -CH3. In some embodiments, -CO-(CH2) m -Z is -CO-(CH2)4-CH3, -CO-(CH2)5-CH3, -CO-(CH2)6-CH3, -CO-(CH2)7-CH3, -CO-(CH2)8-CH3, -CO-(CH2)9-CH3, -CO-(CH2) 10 -CH3, -CO-(CH2) 11 -CH3, -CO-(CH2) 12 -CH3, -CO-(CH2) 13 -CH3, -CO-(CH2) 14 -CH3, -CO-(CH2) 15 -CH3, -CO-(CH2) 16 -CH3, -CO-(CH2) 17 -CH3, -CO-(CH2) 18 -CH3, -CO-(CH2) 19 -CH3 and -CO-(CH2) 20 -Selected from the group consisting of CH3.

[0396] Lipophilic substituents and spacers In some embodiments, the lipophilic substituent is bonded to the parent peptide by a “spacer”. In some embodiments, provided herein is one of the disclosed polypeptides comprising an amino acid sequence selected from the group consisting of amino acid sequences represented by any of the consensus sequences of SEQ ID NOs: 1 to SEQ ID NOs: 143, which include a lipophilic substituent, wherein the lipophilic substituent is bonded to the ε-amino group of lysine via a spacer, which forms a bridge between the amino group of the disclosed polypeptide and the CO-group of the lipophilic substituent.

[0397] In some embodiments, the spacer comprises one or more amino acids, e.g., a single amino acid such as Glu, Asp, Gly, or Lys; a dipeptide such as 2(Glu), Glu-Gly; or a polypeptide such as 3(Glu), 4(Glu), 2(Glu)-Gly. In some embodiments, when the spacer comprises one or more amino acids, e.g., Glu, Asp, Gly, or Lys, one carboxyl group of the spacer can form an amide bond with an amino group of the disclosed polypeptide, and an amino group of the spacer can form an amide bond with a carboxyl group of a lipophilic substituent.

[0398] In some embodiments, if the spacer further comprises Glu or Asp including a carboxylic acid terminal side chain, the terminal carboxyl group of the side chain of the Glu or Asp-containing spacer can form an amide bond with the amino group of the disclosed polypeptide, and the amino group of the Glu or Asp-containing spacer, i.e., γGlu or βAsp, can form an amide bond with the carboxyl group of the lipophilic substituent.

[0399] In some embodiments, the spacer is -γGlu-γGlu-. In some embodiments, the spacer is -γGlu-γGlu-dpeg-. In some embodiments, the spacer is -dpeg-dpeg-γGlu-. In some embodiments, the spacer is -γGlu-dpeg-dpeg-γGlu-. In some embodiments, the spacer is -γGlu-γGlu-dpeg-γGlu-γGlu-. In some embodiments, the spacer is -[COCH2(OCH2CH2)2NH]2-γGlu-.

[0400] In some embodiments, the polypeptide comprises three, two, or preferably one lipophilic substituents, each accompanied by a spacer. In some non-limiting embodiments, the lipophilic substituents and spacers form a monovalent group selected from the group listed in Table 7: [Table 7]

[0401] Preferably, the lipophilic substituent and the spacer are bonded to the amino group of the polypeptide. In particular, the carboxyl group of the lipophilic substituent, or optionally the carboxyl group of the spacer, forms an amide bond with the ε-amino group of the lysine residue. Optionally, the lysine residue bonded to the lipophilic substituent via the spacer may be L-lysine or D-lysine. Typical structural representations of spacer moieties and lipophilic substituents are provided in Table 8: [Table 8-1] [Table 8-2] [Table 8-3]

[0402] In some embodiments, the lipophilic substituent and spacer are of formula II: -(Y)n -CO-(CH2) m -Z Formula II Forms a monovalent group (In the formula, Y is selected from the group consisting of γGlu, Asp, Lys, and Gly; Z is either -CH3 or -CO2H; m is between 4 and 24; n is between 1 and 10.

[0403] In some embodiments, Z is -CO2H. In some embodiments, m is 14-20. In some embodiments, Y is γglu. In some embodiments, n is 1-5.

[0404] In some embodiments, Y is selected from the group consisting of γGlu and Gly. In some embodiments, Y is γGlu. In some embodiments, Y is Gly.

[0405] In some embodiments, the polypeptide comprises three, two, or preferably one lipophilic substituents, each accompanied by a spacer. In some embodiments, the lipophilic substituents and spacers are derived from formula III: -(V) r -(Y) n -CO-(CH2) m -Z Formula III It is a monovalent group. (In the formula, V is -[COCH2(OCH2CH2) t NH]- is; Y is selected from the group consisting of γGlu, Asp, and Gly; Z is either -CH3 or -CO2H; m is between 4 and 24; n is between 1 and 10; r is between 1 and 6; (t is between 1 and 6).

[0406] In some embodiments, Z is -CO2H. In some embodiments, Z is -CH3.

[0407] In some embodiments, Y is γGlu. In some embodiments, Y is Asp. In some embodiments, Y is Gly.

[0408] In some embodiments, m is selected from the group consisting of 4-20, 8-20, 12-20, 14-20, 16-20, 14, 16, 18, and 20. In some embodiments, m is 14-20.

[0409] In some embodiments, n is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. In some embodiments, n is 1 to 5. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

[0410] In some embodiments, r is 1 to 3. In some embodiments, r is 1. In some embodiments, r is 2. In some embodiments, r is 3. In some embodiments, r is 4. In some embodiments, r is 5.

[0411] In some embodiments, t is 1 to 3. In some embodiments, t is selected from the group consisting of 1, 2, 3, 4, 5, and 6.

[0412] In some embodiments, Y is γGlu; ​​Z is -CO2H; m is 16; n is 1; r is 2; and t is 2.

[0413] In one embodiment, -(V) r -(Y) n - is -[COCH2(OCH2CH2)2NH]2-γGlu-.

[0414] In some embodiments, the polypeptide comprises three, two, or preferably one lipophilic substituents, each accompanied by a spacer. In some embodiments, the lipophilic substituents and spacers are of formula IV: -(Y1) n1 -(dpeg) r -(Y2) n2 -CO-(CH2) m -Z Formula IV It is a monovalent group. (In the formula, Z is either -CH3 or -CO2H; m is between 4 and 24; Y1 is selected from the group consisting of γGlu, Asp, and Gly; Y2 is selected from the group consisting of γGlu, Asp, and Gly; dpeg is -[CO(CH2)O(CH2)2O(CH2)NH]-; r is between 1 and 8; n1 is between 0 and 10; n² is between 0 and 10.

[0415] In some embodiments, Z is -CO2H. In some embodiments, Z is -CH3.

[0416] In some embodiments, m is selected from the group consisting of 4-20, 8-20, 12-20, 14-20, 16-20, 14, 16, 18, and 20. In some embodiments, m is 14-20.

[0417] In some embodiments, Y1 is γGlu. In some embodiments, Y1 is Asp. In some embodiments, Y1 is Gly.

[0418] In some embodiments, Y2 is γGlu. In some embodiments, Y2 is Asp. In some embodiments, Y2 is Gly.

[0419] In some embodiments, n1 is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. In some embodiments, n1 is 0 to 3. In some embodiments, n1 is 0. In some embodiments, n1 is 1. In some embodiments, n1 is 2. In some embodiments, n1 is 3. In some embodiments, n1 is 4. In some embodiments, n1 is 5.

[0420] In some embodiments, n2 is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. In some embodiments, n2 is 0 to 3. In some embodiments, n2 is 0. In some embodiments, n2 is 1. In some embodiments, n2 is 2. In some embodiments, n2 is 3. In some embodiments, n2 is 4. In some embodiments, n2 is 5.

[0421] In some embodiments, r is 1 to 3. In some embodiments, r is 1. In some embodiments, r is 2. In some embodiments, r is 3. In some embodiments, r is 4. In some embodiments, r is 5. In some embodiments, r is 6. In some embodiments, r is 7. In some embodiments, r is 8.

[0422] In some embodiments, r is 1, n1 is 2, and n2 is 0.

[0423] In some embodiments, r is 1, n1 is 2, and n2 is 2.

[0424] In some embodiments, Y1 is γGlu and Y2 is γGlu.

[0425] In some embodiments, Y1 is γGlu and n2 is 0.

[0426] In some embodiments, Y1 is γGlu, r is 1, n1 is 2, and n2 is 0.

[0427] In some embodiments, -(Y1) n1 -(dpeg) r -(Y2) n2 - is selected from the group consisting of -γGlu-γGlu-dpeg-, -γGlu-γGlu-dpeg-γGlu-γGlu-, -γGlu-γGlu-dpeg-γGlu-, -γGlu-γGlu-dpeg-dpeg-, -γGlu-γGlu-dpeg-dpeg-γGlu-, -dpeg-dpeg-γGlu-, -γGlu-γGlu-γGlu-dpeg-, and -γGlu-dpeg-.

[0428] In some embodiments, the polypeptide comprises three, two, or preferably one lipophilic substituents, each accompanied by a spacer. In some embodiments, the lipophilic substituents and spacers are of formula V: -(γGlu) n -CO-(CH2) m -Z Formula V It is a monovalent group. (In the formula, Z is either -CH3 or -CO2H; m is between 4 and 24; n is between 1 and 10.

[0429] In some embodiments, Z is -CH3. In some embodiments, Z is -CO2H.

[0430] In some embodiments, m is 14 to 20.

[0431] In some embodiments, m is selected from the group consisting of 4-20, 8-20, 12-20, 14-20, 16-20, 14, 16, 18, and 20.

[0432] In some embodiments, n is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

[0433] In some embodiments, the polypeptide comprises three, two, or preferably one lipophilic substituents, each accompanied by a spacer. In some embodiments, the lipophilic substituents and spacers are of formula VI: -(γGlu) n -(Gly)-CO-(CH2) m -Z Equation VI It is a monovalent group. (In the formula, Z is either -CH3 or -CO2H; m is between 4 and 24; n is between 1 and 10.

[0434] In some embodiments, (γGlu) n This is selected from the group consisting of γGlu; ​​2(γGlu); 3(γGlu); 4(γGlu); and 5(γGlu). In some embodiments, -(γGlu) n -(Gly)- is selected from the group consisting of 2(γGlu),Gly; and 3(γGlu),Gly.

[0435] In some embodiments, the polypeptide comprises three, two, or preferably one lipophilic substituents, each accompanied by a spacer. In some embodiments, the lipophilic substituents and spacers are of formula VII: -(Gly)-(γGlu) n -(CO-(CH2) m -Z Formula VII It is a monovalent group. (In the formula, Z is either -CH3 or -CO2H; m is between 4 and 24; n is between 1 and 10.

[0436] In some embodiments, certain variables in some predicates include:

[0437] In some embodiments, Z is -CH3. In some embodiments, Z is -CO2H.

[0438] In some embodiments, m is selected from the group consisting of 4-20, 8-20, 12-20, 14-20, 16-20, 14, 16, 18, and 20.

[0439] In some embodiments, n is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

[0440] In some embodiments, n is 1 and Z is -CO2H. In some embodiments, n is 1 and Z is -CH3. In some embodiments, n is 2 and Z is -CO2H. In some embodiments, n is 2 and Z is -CH3. In some embodiments, n is 3 and Z is -CO2H. In some embodiments, n is 3 and Z is -CH3. In some embodiments, n is 4 and Z is -CO2H. In some embodiments, n is 4 and Z is -CH3. In some embodiments, n is 5 and Z is -CO2H. In some embodiments, n is 5 and Z is -CH3.

[0441] In some embodiments, n is 1, Z is -CO2H, and m is 14-20. In some embodiments, n is 1, Z is -CO2H, and m is 14. In some embodiments, n is 1, Z is -CO2H, and m is 16. In some embodiments, n is 1, Z is -CO2H, and m is 18.

[0442] In some embodiments, n is 1, Z is -CH3, and m is 14-20. In some embodiments, n is 1, Z is -CH3, and m is 14. In some embodiments, n is 1, Z is -CH3, and m is 16. In some embodiments, n is 1, Z is -CH3, and m is 18.

[0443] In some embodiments, n is 2, Z is -CO2H, and m is 14-20. In some embodiments, n is 2, Z is -CO2H, and m is 14. In some embodiments, n is 2, Z is -CO2H, and m is 16. In some embodiments, n is 2, Z is -CO2H, and m is 18.

[0444] In some embodiments, n is 2, Z is -CH3, and m is 14-20. In some embodiments, n is 2, Z is -CH3, and m is 14. In some embodiments, n is 2, Z is -CH3, and m is 16. In some embodiments, n is 2, Z is -CH3, and m is 18.

[0445] In some embodiments, n is 3, Z is -CO2H, and m is 14-20. In some embodiments, n is 3, Z is -CO2H, and m is 14. In some embodiments, n is 3, Z is -CO2H, and m is 16. In some embodiments, n is 3, Z is -CO2H, and m is 18.

[0446] In some embodiments, n is 3, Z is -CH3, and m is 14-20. In some embodiments, n is 3, Z is -CH3, and m is 14. In some embodiments, n is 3, Z is -CO2H, and m is 16. In some embodiments, n is 3, Z is -CO2H, and m is 18.

[0447] In some embodiments, n is 4, Z is -CO2H, and m is 14-20. In some embodiments, n is 4, Z is -CO2H, and m is 14. In some embodiments, n is 4, Z is -CO2H, and m is 16. In some embodiments, n is 4, Z is -CO2H, and m is 18.

[0448] In some embodiments, n is 4, Z is -CH3, and m is 14-20. In some embodiments, n is 4, Z is -CH3, and m is 14. In some embodiments, n is 4, Z is -CH3, and m is 16. In some embodiments, n is 4, Z is -CH3, and m is 18.

[0449] In some embodiments, n is 5, Z is -CO2H, and m is 14-20. In some embodiments, n is 5, Z is -CO2H, and m is 14. In some embodiments, n is 5, Z is -CO2H, and m is 16. In some embodiments, n is 5, Z is -CO2H, and m is 18.

[0450] In some embodiments, n is 5, Z is -CH3, and m is 14-20. In some embodiments, n is 5, Z is -CH3, and m is 14. In some embodiments, n is 5, Z is -CH3, and m is 16. In some embodiments, n is 5, Z is -CH3, and m is 18.

[0451] In some embodiments, the polypeptide comprises three, two, or preferably one lipophilic substituents, each accompanied by a spacer. In some embodiments, the lipophilic substituents and spacers are of formula VIII: -(Y1) n1 -(V) r -(Y2) n2 -CO-(CH2) m -Z Formula VIII It is a monovalent group. (In the formula, Z is either -CH3 or -CO2H; m is between 4 and 24; Y1 is selected from the group consisting of γGlu, Asp, and Gly; Y2 is selected from the group consisting of γGlu, Asp, and Gly; V is -[COCH2(OCH2CH2) t NH]- is; r is between 1 and 6; n1 is between 0 and 10; n² is between 0 and 10; (t is between 1 and 6).

[0452] In some embodiments, the polypeptide comprises three, two, or preferably one lipophilic substituents, each accompanied by a spacer. In some embodiments, the lipophilic substituents and spacers are of formula IX: -(Y) n -(V) r -CO-(CH2) m -Z The monovalent group in formula IX: (In the formula, Z is either -CH3 or -CO2H; m is between 4 and 24; Y is selected from the group consisting of γGlu, Asp, and Gly; V is -[COCH2(OCH2CH2) t NH]- is; r is between 1 and 6; n is between 1 and 10; (t is between 1 and 6).

[0453] In some embodiments, the polypeptide comprises three, two, or preferably one lipophilic substituents, each accompanied by a spacer. In some embodiments, the lipophilic substituents and spacers are of formula X: -(dpeg) r -(Y2) n2 -CO-(CH2) m -Z formula It is a monovalent group. (In the formula, Z is either -CH3 or -CO2H; m is between 4 and 24; dpeg is -[CO(CH2)O(CH2)2O(CH2)NH]-; Y2 is selected from the group consisting of γGlu, Asp, and Gly; r is between 1 and 8; n² is between 0 and 10.

[0454] In some embodiments, -(dpeg) r -(Y2) n2 - is selected from the group consisting of dpeg,γGlu; ​​and dpeg,dpeg,γGlu.

[0455] In one embodiment, -(dpeg) r -(Y2) n2 - is -dpeg-dpeg-γGlu-.

[0456] In some embodiments, the polypeptide comprises three, two, or preferably one lipophilic substituents, each accompanied by a spacer. In some embodiments, the lipophilic substituents and spacers are of formula XI: -(Y1) n1 -(dpeg) r -CO-(CH2) m -Z Formula XI It is a monovalent group. (In the formula, Z is either -CH3 or -CO2H; m is between 4 and 24; Y1 is selected from the group consisting of γGlu, Asp, and Gly; dpeg is -[CO(CH2)O(CH2)2O(CH2)NH]-; r is between 1 and 8; n1 is between 0 and 10.

[0457] In some embodiments, Z is -CO2H.

[0458] In some embodiments, m is selected from the group consisting of 4-20, 8-20, 12-20, 14-20, 16-20, 14, 16, 18, and 20. In some embodiments, m is 14-20.

[0459] In some embodiments, Y1 is γGlu. In some embodiments, Y1 is Asp. In some embodiments, Y1 is Gly.

[0460] In some embodiments, n1 is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. In some embodiments, n1 is 0 to 3. In some embodiments, n1 is 0. In some embodiments, n1 is 1. In some embodiments, n1 is 2. In some embodiments, n1 is 3. In some embodiments, n1 is 4. In some embodiments, n1 is 5.

[0461] In some embodiments, r is 1 to 3. In some embodiments, r is 1. In some embodiments, r is 2. In some embodiments, r is 3. In some embodiments, r is 4. In some embodiments, r is 5. In some embodiments, r is 6. In some embodiments, r is 7. In some embodiments, r is 8.

[0462] In some embodiments, r is 1 and n1 is 2.

[0463] In some embodiments, Y1 is γGlu, r is 1, and n1 is 2.

[0464] In some embodiments, -(Y1) n1 -(dpeg) r - is selected from the group consisting of -γGlu-γGlu-dpeg-, -γGlu-γGlu-dpeg-dpeg-, -γGlu-γGlu-γGlu-dpeg-, and -γGlu-dpeg-.

[0465] Further exemplary spacers In some embodiments, the spacer is given by formula XII: -N(R1)(CHR2) p CO-[N(R3)((CH2)2O(CH2)2O) q (CH2)CO-] r Formula XII Contains a divalent group (In the formula, Each of R1 and R3 is either hydrogen or a C1-C4 alkyl group; Each R2 is either H or CO2H; p is 1, 2, 3, 4, 5, or 6. q is 1, 2, or 3; r is either 0 or 1. This spacer forms a crosslink between the amino group of the disclosed polypeptide and the CO-group of the lipophilic substituent.

[0466] In some embodiments, the spacer is given by formula XIII: [-N(R3)((CH2)2O(CH2)2O) q (CH2)CO-] r Formula XIII Contains a divalent group (In the formula, R3 is hydrogen or a C1-C4 alkyl group; q is 1, 2, or 3; r is either 0 or 1. This spacer forms a crosslink between the amino group of the disclosed polypeptide and the CO-group of the lipophilic substituent.

[0467] In some embodiments, a particular variable represented by a particular expression includes the following: In some embodiments, each R1 is hydrogen. In some embodiments, each R3 is hydrogen. In some embodiments, each R1 and each R3 are hydrogen.

[0468] In some embodiments, at least one R2 is CO2H.

[0469] In some embodiments, p is 1. In some embodiments, p is 2. In some embodiments, p is 3. In some embodiments, p is 4. In some embodiments, p is 5. In some embodiments, p is 6.

[0470] In some embodiments, q is 1. In some embodiments, q is 2. In some embodiments, q is 3.

[0471] In some embodiments, r is 0. In some embodiments, r is 1.

[0472] In some embodiments, the spacer is γ-glutamyl, i.e., -NH(CHCO2H)(CH2)2CO--. In some embodiments, the spacer is γ-aminobutanoyl, i.e., -NH(CH2)3CO--. In some embodiments, the spacer is β-asparagyl, i.e., -NH(CHCO2H)(CH2)CO--. In some embodiments, the spacer is -NH(CH2)2CO--. In some embodiments, the spacer is glycyl. In some embodiments, the spacer is β-alanyl.

[0473] In some embodiments, the spacer is -NHCH(CO2H)(CH2)2CO--[N(R3)((CH2)2O(CH2)2O) q (CH2)CO-] r In some embodiments, the spacer is -NH(CH2)3CO--[N(R3)((CH2)2O(CH2)2O) q (CH2)CO-] r In some embodiments, the spacer is -NHCH(CO2H)(CH2)2CO-NH((CH2)2O(CH2)2O)2(CH2)CO-. In some embodiments, the spacer is -NH(CH2)3CO-NH((CH2)2O(CH2)2O)2(CH2)CO-. In some embodiments, the spacer is -NHCH(CO2H)CH2CO--[N(R3)((CH2)2O(CH2)2O) q (CH2)CO-] r In some embodiments, the spacer is -NH(CH2)2CO--[N(R3)((CH2)2O(CH2)2O) q (CH2)CO-] r That is the case.

[0474] In some embodiments, the spacer is given by formula XIV: -(Y) n - Formula XIV Contains a divalent group (In the formula, Y is selected from the group consisting of γGlu, Asp, Lys, and Gly; n is between 1 and 10.

[0475] In some embodiments, Y is selected from the group consisting of γGlu and Gly. In some embodiments, Y is γGlu. In some embodiments, Y is Gly.

[0476] In some embodiments, the spacer forms a crosslink between the amino group of the disclosed polypeptide and the CO- group of the lipophilic substituent. In some embodiments, one end of the spacer forms a covalent bond with the amino group of the disclosed polypeptide, and the other end of the spacer forms a covalent bond with a hydrogen atom or a protecting group.

[0477] In some embodiments, the spacer is given by formula XV: -(γGlu) n - Formula XV Contains a divalent group (In the formula, n is between 1 and 10. In some embodiments, n is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

[0478] In some embodiments, the spacer forms a crosslink between the amino group of the disclosed polypeptide and the CO- group of the lipophilic substituent. In some embodiments, one end of the spacer forms a covalent bond with the amino group of the disclosed polypeptide, and the other end of the spacer forms a covalent bond with a hydrogen atom or a protecting group.

[0479] In some embodiments, the spacer is given by formula XVI: -(γGlu) n -(Gly)- Formula XVI It contains a divalent group. n is between 1 and 10. In some embodiments, (γGlu) n This is selected from the group consisting of γGlu; ​​2(γGlu); 3(γGlu); 4(γGlu); and 5(γGlu). In some embodiments, -(γGlu) n-(Gly)- is selected from the group consisting of 2(γGlu),Gly; and 3(γGlu),Gly. In some embodiments, the spacer forms a bridge between the amino group of the disclosed polypeptide and the CO- group of the lipophilic substituent. In some embodiments, one end of the spacer forms a covalent bond with the amino group of the disclosed polypeptide, and the other end of the spacer forms a covalent bond with a hydrogen atom or a protecting group.

[0480] In some embodiments, the spacer is given by formula XVII: -(Gly)-(γGlu) n - Formula XVII Contains a divalent group (In the formula, n is between 1 and 10. In some embodiments, the spacer forms a crosslink between the amino group of the disclosed polypeptide and the CO- group of the lipophilic substituent. In some embodiments, one end of the spacer forms a covalent bond with the amino group of the disclosed polypeptide, and the other end of the spacer forms a covalent bond with a hydrogen atom or a protecting group.

[0481] In some embodiments, n is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

[0482] In some embodiments, the spacer is given by formula XVIII: -(V) r -(Y) n - Formula XVIII Contains a divalent group (In the formula, Y is selected from the group consisting of γGlu, Asp, and Gly; V is -[COCH2(OCH2CH2) t NH]- is; r is between 1 and 6; n is between 1 and 10; (t is between 1 and 6).

[0483] In some embodiments, Y is γGlu. In some embodiments, Y is Asp. In some embodiments, Y is Gly.

[0484] In some embodiments, n is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. In some embodiments, n is 1 to 5. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.

[0485] In some embodiments, r is 1 to 3. In some embodiments, r is 1. In some embodiments, r is 2. In some embodiments, r is 3. In some embodiments, r is 4. In some embodiments, r is 5.

[0486] In some embodiments, t is 1 to 3. In some embodiments, t is selected from the group consisting of 1, 2, 3, 4, 5, and 6.

[0487] In one embodiment, -(V) r -(Y) n - is -[COCH2(OCH2CH2)2NH]2-γGlu-.

[0488] In some embodiments, the spacer forms a crosslink between the amino group of the disclosed polypeptide and the CO- group of the lipophilic substituent. In some embodiments, one end of the spacer forms a covalent bond with the amino group of the disclosed polypeptide, and the other end of the spacer forms a covalent bond with a hydrogen atom or a protecting group.

[0489] In some embodiments, the spacer is represented by formula XIX: -(Y1) n1 -(dpeg) r -(Y2) n2- Formula XIX Contains a divalent group (In the formula, Y1 is selected from the group consisting of γGlu, Asp, and Gly; Y2 is selected from the group consisting of γGlu, Asp, and Gly; dpeg is -[CO(CH2)O(CH2)2O(CH2)NH]-; r is between 1 and 8; n1 is between 0 and 10; n² is between 0 and 10.

[0490] In some embodiments, Y1 is γGlu. In some embodiments, Y1 is Asp. In some embodiments, Y1 is Gly.

[0491] In some embodiments, Y2 is γGlu. In some embodiments, Y2 is Asp. In some embodiments, Y2 is Gly.

[0492] In some embodiments, n1 is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. In some embodiments, n1 is 0 to 3. In some embodiments, n1 is 0. In some embodiments, n1 is 1. In some embodiments, n1 is 2. In some embodiments, n1 is 3. In some embodiments, n1 is 4. In some embodiments, n1 is 5.

[0493] In some embodiments, n2 is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. In some embodiments, n2 is 0 to 3. In some embodiments, n2 is 0. In some embodiments, n2 is 1. In some embodiments, n2 is 2. In some embodiments, n2 is 3. In some embodiments, n2 is 4. In some embodiments, n2 is 5.

[0494] In some embodiments, r is 1 to 3. In some embodiments, r is 1. In some embodiments, r is 2. In some embodiments, r is 3. In some embodiments, r is 4. In some embodiments, r is 5. In some embodiments, r is 6. In some embodiments, r is 7. In some embodiments, r is 8.

[0495] In some embodiments, r is 1, n1 is 2, and n2 is 0.

[0496] In some embodiments, r is 1, n1 is 2, and n2 is 2.

[0497] In some embodiments, Y1 is γGlu and Y2 is γGlu.

[0498] In some embodiments, Y1 is γGlu and n2 is 0.

[0499] In some embodiments, Y1 is γGlu, r is 1, n1 is 2, and n2 is 0.

[0500] In some embodiments, -(Y1) n1 -(dpeg) r -(Y2) n2 - is selected from the group consisting of -γGlu-γGlu-dpeg-, -γGlu-γGlu-dpeg-γGlu-γGlu-, -γGlu-γGlu-dpeg-γGlu-, -γGlu-γGlu-dpeg-dpeg-, -γGlu-γGlu-dpeg-dpeg-γGlu-, -dpeg-dpeg-γGlu-, -γGlu-γGlu-γGlu-dpeg-, and -γGlu-dpeg-.

[0501] In some embodiments, the spacer forms a crosslink between the amino group of the disclosed polypeptide and the CO- group of the lipophilic substituent. In some embodiments, one end of the spacer forms a covalent bond with the amino group of the disclosed polypeptide, and the other end of the spacer forms a covalent bond with a hydrogen atom or a protecting group.

[0502] Accordingly, in some embodiments, the isolated polypeptides provided herein comprise an amino acid sequence selected from the group comprising an amino acid sequence represented by a consensus sequence selected from the group consisting of SEQ ID NOs: 101 to 181 and SEQ ID NOs: 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 280, and 281, or a pharmaceutically acceptable salt thereof, wherein the isolated peptide further comprises lipophilic substituents and optionally comprises spacers.

[0503] In some embodiments, the isolated polypeptides provided herein comprise an amino acid sequence selected from the group comprising an amino acid sequence represented by a consensus sequence selected from the group consisting of SEQ ID NOs: 101 to 181 and SEQ ID NOs: 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 280, and 281, or a pharmaceutically acceptable salt thereof, wherein the isolated peptide further comprises a lipophilic substituent of formula I.

[0504] In some embodiments, the isolated polypeptides provided herein comprise an amino acid sequence selected from the group comprising an amino acid sequence represented by a consensus sequence selected from the group comprising SEQ ID NOs: 101 to 181 and SEQ ID NOs: 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 280, and 281, or a pharmaceutically acceptable salt thereof, wherein the isolated peptide further comprises a lipophilic substituent of formula I and a spacer selected from the group comprising those described by formulas XII, XIII, XIV, XV, XVI, XVII, XVIII, and XIX.

[0505] In some embodiments, the isolated polypeptides provided herein comprise an amino acid sequence selected from the group comprising an amino acid sequence represented by a consensus sequence selected from the group comprising SEQ ID NOs: 101 to 157 and SEQ ID NOs: 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 280, and 281, or a pharmaceutically acceptable salt thereof, wherein the isolated peptide further comprises lipophilic substituents and spacers selected from the group comprising those described by formulas I, II, III, IV, V, VI, VII, VIII, IX, X, and XI.

[0506] In some embodiments, the isolated polypeptides provided herein comprise an amino acid sequence selected from the group comprising an amino acid sequence represented by a consensus sequence selected from the group comprising SEQ ID NOs: 101 to 181 and SEQ ID NOs: 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 280, and 281, or a pharmaceutically acceptable salt thereof, wherein the isolated peptide further comprises lipophilic substituents and spacers selected from the group comprising those described by Formula I.

[0507] In some embodiments, the isolated polypeptides provided herein comprise an amino acid sequence selected from the group comprising an amino acid sequence represented by a consensus sequence selected from the group comprising SEQ ID NOs: 101 to 181 and SEQ ID NOs: 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 280, and 281, or a pharmaceutically acceptable salt thereof, wherein the isolated peptide further comprises lipophilic substituents and spacers selected from the group comprising those described by Formula II.

[0508] In some embodiments, the isolated polypeptides provided herein comprise an amino acid sequence selected from the group comprising an amino acid sequence represented by a consensus sequence selected from the group comprising SEQ ID NOs: 101 to 181 and SEQ ID NOs: 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 280, and 281, or a pharmaceutically acceptable salt thereof, wherein the isolated peptide further comprises lipophilic substituents and spacers selected from the group comprising those described by Formula III.

[0509] In some embodiments, the isolated polypeptides provided herein comprise an amino acid sequence selected from the group comprising an amino acid sequence represented by a consensus sequence selected from the group comprising SEQ ID NOs: 101 to 181 and SEQ ID NOs: 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 280, and 281, or a pharmaceutically acceptable salt thereof, wherein the isolated peptide further comprises lipophilic substituents and spacers selected from the group comprising those described by Formula IV. As used herein, (γGlu)2 and 2(γGlu) both mean -(γGlu)-(γGlu)- or -CO(CH2)2CH(CO2H)NH-CO(CH2)2CH(CO2H)NH-; (γGlu)3 and 3(γGlu) both mean -(γGlu)-(γGlu)-(γGlu)- or -CO(CH2)2CH(CO2H)NH-CO(CH2)2CH(CO2H)NH-CO(CH2)2CH(CO2H)NH-; and so on; if a variable appears multiple times in a given formula, each occurrence of that variable is determined independently. For example, for a -(Y)3- group where Y can be γGlu, Asp, Lys, or Gly, each Y is independently selected such that it is one of four amino acids. Therefore, according to the non-restrictive example, -(Y)3- could be -(γGlu)-(γGlu)-(γGlu)-, -(γGlu)-(Asp)-(γGlu)-, -(Gly)-(Asp)-(γGlu)-, or -(Gly)-(γGlu)-(γGlu)-.

[0510] bridging part In some embodiments, any of the disclosed polypeptides are optionally substituted with one or more crosslinking moieties. As used herein, the term “crosslinking moiety” means a covalent bond or any divalent linker or a moiety that links two side chains of two distinct amino acid residues. In some embodiments, any of the disclosed polypeptides are optionally substituted with one or more lactam crosslinking moieties. As used herein, the term “lactam crosslinking moiety” means a lactam crosslink or lactam bond that links an amino-containing side chain and a carboxy-containing side chain of two distinct amino acid residues. In some embodiments, a lactam crosslinking moiety is formed between a lysine residue and an aspartic acid residue, where the amino-containing side chain of lysine and the carboxy-containing side chain of aspartic acid covalently bond by losing water to form a lactam crosslinking moiety. In some embodiments, a lactam crosslinking moiety is formed between a lysine residue and a glutamic acid residue, where the amino-containing side chain of lysine and the carboxy-containing side chain of glutamic acid covalently bond by losing water to form a lactam crosslinking moiety. In some embodiments, the lactam crosslinking moiety is formed between two amino acids that are 3, 4, or 5 residues apart on the peptide. In some embodiments, the lactam crosslinking moiety is formed between two amino acids that are 4 residues apart on the peptide.

[0511] General method for providing polypeptide intermediates and the compound. The compounds of the present invention can generally be prepared or isolated by synthetic and / or semi-synthetic methods known to those skilled in the art for similar compounds, and by methods described in detail in the examples herein.

[0512] Where a specific protecting group ("PG"), leaving group ("LG"), or conversion condition is shown in the following scheme, those skilled in the art will understand that other protecting groups, leaving groups, and conversion conditions are also suitable and intended. For such groups and conversions, see March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, MB Smith and J. March, 5 th Edition, John Wiley & Sons, 2001, Comprehensive Organic Transformations, RC Larock, 2 nd Edition, John Wiley & Sons, 1999, and Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3 rd This is described in detail in the edition, John Wiley & Sons, 1999, and each of these in their entirety is incorporated herein by reference.

[0513] As used herein, the term “leaving group” (LG) includes, but is not limited to, halogens (e.g., fluorides, chlorides, bromides, iodides), sulfonates (e.g., mesylates, tosylates, benzenesulfonates, brosylates, nosylates, triflates), diazoniums, and others.

[0514] As used herein, the phrase “oxygen protecting group” includes, for example, carbonyl protecting groups and hydroxyl protecting groups. Hydroxyl protecting groups are well known in the art, as seen in Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3. rdThis includes the details described in John Wiley & Sons, 1999 edition, which is incorporated herein by reference. Suitable examples of hydroxyl protecting groups include, but are not limited to, esters (e.g., acetyl, benzyl), allyl ethers, ethers, silyl ethers, alkyl ethers, arylalkyl ethers, benzyl ethers, and alkoxyalkyl ethers. Examples of such esters include formic acid esters, acetate esters, carbonate esters, and sulfonic acid esters. Specific examples include formic acid esters, benzoyl formates, chloroacetates, trifluoroacetates, methoxyacetates, triphenylmethoxyacetates, p-chlorophenoxyacetates, 3-phenylpropionate, 4-oxopentanoate, 4,4-(ethylenedithio)pentanoate, pivaloate (trimethylacetyl), crotonate, 4-methoxycrotonate, benzoic acid esters, p-benzylbenzoate, 2,4,6-trimethylbenzoate, carbonate esters, such as methyl, 9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl, 2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl, vinyl, allyl, and p-nitrobenzyl. Examples of such silyl ethers include trimethylsilyl ether, triethylsilyl ether, t-butyldimethylsilyl ether, t-butyldiphenylsilyl ether, triisopropylsilyl ether, and other trialkylsilyl ethers. Examples of alkyl ethers include methyl ether, benzyl ether, p-methoxybenzyl ether, 3,4-dimethoxybenzyl ether, trityl ether, t-butyl ether, allyl ether, and allyloxycarbonyl ether or derivatives. Examples of alkoxyalkyl ethers include acetals, such as methoxymethyl ether, methylthiomethyl ether, (2-methoxyethoxy)methyl ether, benzyloxymethyl ether, beta-(trimethylsilyl)ethoxymethyl ether, and tetrahydropyranyl ether.Examples of arylalkyl ethers include benzyl, p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, and 2- and 4-picolyl.

[0515] Amino protecting groups are well known in the art, as seen in Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3 rd This includes those described in detail in edition, John Wiley & Sons, 1999, which is incorporated herein by reference. Suitable amino protecting groups include, but are not limited to, aralkylamines, carbamates, cyclic imides, allylamines, and amides. Examples of such groups include t-butyloxycarbonyl (Boc), ethyloxycarbonyl, methyloxycarbonyl, trichloroethyloxycarbonyl, allyloxycarbonyl (Alloc), benzyl oxocarbonyl (Cbz), allyl, phthalimide, benzyl (Bn), dimethyl-2,6-dioxocyclohexa-1-ylidene)ethyl (Dmb), 1-(4,4-dimethyl-2,6-dioxocyclohexa-1-ylidene)-3-methylbutyl (ivDde), fluorenylmethylcarbonyl (Fmoc), formyl, acetyl, chloroacetyl, dichloroacetyl, trichloroacetyl, phenylacetyl, trifluoroacetyl, and benzoyl.

[0516] In certain embodiments, the present invention also relates to synthetic peptide intermediates of the disclosed PYY analogs. In some embodiments, the polypeptide intermediates of the present disclosure are isolated polypeptides comprising any of the amino acid sequences of SEQ ID NOs: 101 to 181, or SEQ ID NOs: 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 280, and 281, wherein at least one amino acid is covalently bonded to a protecting group. In some embodiments, the polypeptide intermediates of the present disclosure are isolated polypeptides comprising any of the amino acid sequences of SEQ ID NOs: 101 to 181, or SEQ ID NOs: 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 280, and 281, wherein at least one amino acid is covalently bonded to a spacer as defined herein, the spacer being further covalently bonded to a protecting group or a hydrogen atom. In some embodiments, the polypeptide intermediate comprises a lysine residue bonded to a protecting group via an amino group of its side chain. In some embodiments, the lysine residue is covalently bonded to Alloc or ivDde. In some embodiments, the polypeptide intermediate includes an aspartic acid residue bonded to a protecting group via a carboxyl group in its side chain. In some embodiments, the aspartic acid residue is covalently bonded to an allyl group. In some embodiments, the polypeptide intermediate includes a glutamic acid residue bonded to a protecting group via a carboxyl group in its side chain. In some embodiments, the glutamic acid residue is covalently bonded to an allyl group.

[0517] Exemplary polypeptide intermediates The compounds of the present invention can generally be prepared or isolated by synthetic and / or semi-synthetic methods known to those skilled in the art for similar compounds, and by methods described in detail in the examples herein.

[0518] In certain embodiments, the present invention also relates to synthetic peptide intermediates of the disclosed PYY analogs. In some embodiments, the polypeptide intermediate of the present disclosure is SEQ ID NO: 390: X0PX2PX4X5PX7X8X9X 10 SPX13 X 14 X 15 X 16 RX 18 X 19 X 20 DX 22 X 23 HX 25 X 26 X 27 WLTRX 32 RX 34 -(OH / NH2)(SEQ ID NO: 390) is an isolated polypeptide containing the amino acid sequence, or a pharmaceutically acceptable salt thereof (wherein the formula, X0 is either nonexistent or K; X2 is K; X4 is either E or K; X5 is either A or K; X7 is either G or K; X8 is E, K, or k; X9 is either D or K; X 10 is A or K; X 13 is either E or K; X 14 is either E or K; X 15 is L or W; X 16 These are D, E, K, N, Q, S, T, α-methylserine, or homoserine; X 18 is either K or Y; X 19 is either K or Y; X 20 is A, D, E, K, k, or Dap; X 22 is A, D, K, or L; X 23 is either K or R; X 25 is either K or Y; X 26 is E, K, or L; X 27 is either K or N; X32 is either K or Q; X 34 These are F, y, 3-pyridinylalanine, 4-pyridinylalanine, 4-carboxyphenylalanine, 4-fluorophenylalanine, 4-methylphenylalanine, N-methylphenylalanine, homophenylalanine, β-homotyrosine, homotyrosine, or N-methyltyrosine; Here, X0, X2, X4, X5, X7, X8, X 13 , X 20 , X 23 , X 25 , X 27 , or X 32 When it is K, the lysine residue is optionally covalently bonded to the protecting group or a spacer optionally bonded to the protecting group. Here, X8 or X 20 If the value is k, the D-lysine residue is optionally covalently bonded to a protecting group or a spacer optionally bonded to a protecting group.

[0519] In some embodiments, X 15 If X is L, 22 It is A.

[0520] In some embodiments, X 16 If X is N, 20 It is not A.

[0521] In some embodiments, X8 is E, X5 is K, and X 20 K is K.

[0522] In some embodiments, X8 is K. In some embodiments, X8 is K covalently bonded to a protecting group. In some embodiments, X8 is K covalently bonded to a spacer optionally bonded to a protecting group.

[0523] In some embodiments, X 15 It is W.

[0524] In some embodiments, X34 It is F.

[0525] In some embodiments, X0 is absent. In some embodiments, X0 is K. In some embodiments, X0 is K covalently bonded to a protecting group. In some embodiments, X0 is K covalently bonded to a spacer optionally bonded to a protecting group.

[0526] In some embodiments, X2 is K. In some embodiments, X2 is K covalently bonded to a protecting group. In some embodiments, X2 is K covalently bonded to a spacer optionally bonded to a protecting group.

[0527] In some embodiments, X4 is E. In some embodiments, X4 is K. In some embodiments, X4 is K covalently bonded to a protecting group. In some embodiments, X4 is K covalently bonded to a spacer optionally bonded to a protecting group.

[0528] In some embodiments, X5 is A. In some embodiments, X5 is K. In some embodiments, X5 is K covalently bonded to a protecting group. In some embodiments, X5 is K covalently bonded to a spacer optionally bonded to a protecting group.

[0529] In some embodiments, X7 is G. In some embodiments, X7 is K. In some embodiments, X7 is K covalently bonded to a protecting group. In some embodiments, X7 is K covalently bonded to a spacer optionally bonded to a protecting group.

[0530] In some embodiments, X8 is E. In some embodiments, X8 is K. In some embodiments, X8 is K covalently bonded to the protecting group. In some embodiments, X8 is K covalently bonded to a spacer optionally bonded to the protecting group. In some embodiments, X8 is k. In some embodiments, X8 is k covalently bonded to the protecting group. In some embodiments, X8 is k covalently bonded to a spacer optionally bonded to the protecting group.

[0531] In some embodiments, X9 is D. In some embodiments, X9 is K.

[0532] In some embodiments, X 10 In some embodiments, X 10 In some embodiments, X 10 is K covalently bonded to the protecting group. In some embodiments, X 10 This is K covalently bonded to a spacer that is optionally bonded to a protecting group.

[0533] In some embodiments, X 13 In some embodiments, X 13 In some embodiments, X 13 is K covalently bonded to the protecting group. In some embodiments, X 13 This is K covalently bonded to a spacer that is optionally bonded to a protecting group.

[0534] In some embodiments, X 14 In some embodiments, X 14 K is K.

[0535] In some embodiments, X 15 L is. In some embodiments, X 15 It is W.

[0536] In some embodiments, X 16In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 Q is Q. In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 X is α-methylserine. In some embodiments, X 16 It is homoserine.

[0537] In some embodiments, X 18 In some embodiments, X 18 Y is Y.

[0538] In some embodiments, X 19 In some embodiments, X 19 Y is Y.

[0539] In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 is K covalently bonded to the protecting group. In some embodiments, X 20 This is K covalently bonded to a spacer optionally bonded to a protecting group. In some embodiments, X 20 is k. In some embodiments, X 20 is k covalently bonded to the protecting group. In some embodiments, X 20 k is covalently bonded to a spacer optionally bonded to a protecting group. In some embodiments, X 20 It is a DAP.

[0540] In some embodiments, X 22In some embodiments, X 22 In some embodiments, X 22 In some embodiments, X 22 It is L.

[0541] In some embodiments, X 23 In some embodiments, X 23 is K covalently bonded to the protecting group. In some embodiments, X 23 This is K covalently bonded to a spacer optionally bonded to a protecting group. In some embodiments, X 23 This is R.

[0542] In some embodiments, X 25 In some embodiments, X 25 is K covalently bonded to the protecting group. In some embodiments, X 25 This is K covalently bonded to a spacer optionally bonded to a protecting group. In some embodiments, X 25 Y is Y.

[0543] In some embodiments, X 26 In some embodiments, X 26 In some embodiments, X 26 It is L.

[0544] In some embodiments, X 27 In some embodiments, X 27 is K covalently bonded to the protecting group. In some embodiments, X 27 This is K covalently bonded to a spacer optionally bonded to a protecting group. In some embodiments, X 27 It is N.

[0545] In some embodiments, X 32 In some embodiments, X 32 is K covalently bonded to the protecting group. In some embodiments, X 32This is K covalently bonded to a spacer optionally bonded to a protecting group. In some embodiments, X 32 Q is Q.

[0546] In some embodiments, X 34 In some embodiments, X 34 y is. In some embodiments, X 34 is 3-pyridinylalanine. In some embodiments, X 34 is 4-pyridinylalanine. In some embodiments, X 34 X is 4-carboxyphenylalanine. In some embodiments, X 34 X is 4-fluorophenylalanine. In some embodiments, X 34 X is 4-methylphenylalanine. In some embodiments, X 34 is N-methylphenylalanine. In some embodiments, X 34 X is homophenylalanine. In some embodiments, X 34 is β-homotyrosine. In some embodiments, X 34 is homotyrosine. In some embodiments, X 34 It is N-methyltyrosine.

[0547] In some embodiments, the polypeptide intermediate of this disclosure is SEQ ID NO: 394: PKPEX5PGX8DASPX 13 EW Test 16 RYYX 20 DX 22 RHYLNWLTRQRX 34 An isolated polypeptide containing -(OH / NH2)(SEQ ID NO: 394), or a pharmaceutically acceptable salt thereof. (In the formula, X5 is either A or K; X8 is E, K, or k; X 13 is either E or K; X 16 is D, E, K, or N; X 20 is A, D, E, K, or k; X 22 is A or L; X 34 is F or N-methyltyrosine; Here, X5, X8, X 13 , or X 20 When it is K, the lysine residue is optionally covalently bonded to the protecting group or a spacer optionally bonded to the protecting group. Here, X8 or X 20 If the value is k, the D-lysine residue is optionally covalently bonded to the protecting group or a spacer optionally bonded to the protecting group.

[0548] In some embodiments, X 16 If X is N, 20 It is not A.

[0549] In some embodiments, X8 is E, X5 is K, and X 20 K is K.

[0550] In some embodiments, X8 is K. In some embodiments, X8 is K covalently bonded to a protecting group. In some embodiments, X8 is K covalently bonded to a spacer optionally bonded to a protecting group.

[0551] In some embodiments, X 34 It is F.

[0552] In some embodiments, X5 is A. In some embodiments, X5 is K. In some embodiments, X5 is K covalently bonded to a protecting group. In some embodiments, X5 is K covalently bonded to a spacer optionally bonded to a protecting group.

[0553] In some embodiments, X8 is E. In some embodiments, X8 is K. In some embodiments, X8 is K covalently bonded to the protecting group. In some embodiments, X8 is K covalently bonded to a spacer optionally bonded to the protecting group. In some embodiments, X8 is k. In some embodiments, X8 is k covalently bonded to the protecting group. In some embodiments, X8 is k covalently bonded to a spacer optionally bonded to the protecting group.

[0554] In some embodiments, X 13 In some embodiments, X 13 In some embodiments, X 13 is K covalently bonded to the protecting group. In some embodiments, X 13 This is K covalently bonded to a spacer that is optionally bonded to a protecting group.

[0555] In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 In some embodiments, X 16 It is N.

[0556] In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 In some embodiments, X 20 is K covalently bonded to the protecting group. In some embodiments, X 20 This is K covalently bonded to a spacer optionally bonded to a protecting group. In some embodiments, X 20 is k. In some embodiments, X 20 is k covalently bonded to the protecting group. In some embodiments, X 20 k is covalently bonded to a spacer that is optionally bonded to a protecting group.

[0557] In some embodiments, X 22 In some embodiments, X 22 It is L.

[0558] In some embodiments, X 34 In some embodiments, X 34 It is N-methyltyrosine.

[0559] In some embodiments, the isolated polypeptides of this disclosure include an amino acid sequence selected from the group consisting of the following peptides listed in Tables 9, 10, 11, and 12. [ka] [ka] [Table 9]

[0560] In some embodiments, the present invention provides peptide intermediates of compound A24 (SEQ ID NO: 24). In some embodiments, the peptide intermediate of compound A24 comprises at least one amino acid covalently bonded to a protecting group. In some embodiments, the peptide intermediate of compound A24 comprises a lysine residue bonded to the protecting group via an amino group of its side chain. In some embodiments, the lysine residue is covalently bonded to Alloc or ivDde. In some embodiments, the peptide intermediate of compound A24 comprises an aspartic acid residue bonded to the protecting group via a carboxyl group of its side chain. In some embodiments, the aspartic acid residue is covalently bonded to an allyl group. In some embodiments, the peptide intermediate of compound A24 comprises a glutamic acid residue bonded to the protecting group via a carboxyl group of its side chain. In some embodiments, the glutamic acid residue is covalently bonded to an allyl group. In some embodiments, the peptide intermediate of compound A24 comprises at least one amino acid covalently bonded to a spacer as defined herein. The spacer is further covalently bonded to a protecting group or a hydrogen atom. In some embodiments, the present invention provides peptide intermediates shown in Table 10 above. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 324. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 424. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 124. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 524.

[0561] In some embodiments, the present invention relates to a method for preparing compounds of the present invention, such as compound A24 (SEQ ID NO: 24), wherein a polypeptide intermediate such as compound D24 (SEQ ID NO: 324) is activated by the following activated acyl group, namely (LG)CO(CH2), as illustrated in Scheme 1 below. zz The present invention provides a method comprising the step of acylation with CO2H, wherein ZZ is 14-22 and LG is a leaving group as defined herein:

[0562] [Table 10] [Table 11]

[0563] In some embodiments, the present invention provides peptide intermediates of compound A42 (SEQ ID NO: 42). In some embodiments, the peptide intermediate of compound A42 comprises at least one amino acid covalently bonded to a protecting group. In some embodiments, the peptide intermediate of compound A42 comprises a lysine residue bonded to the protecting group via an amino group of its side chain. In some embodiments, the lysine residue is covalently bonded to Alloc or ivDde. In some embodiments, the peptide intermediate of compound A42 comprises an aspartic acid residue bonded to the protecting group via a carboxyl group of its side chain. In some embodiments, the aspartic acid residue is covalently bonded to an allyl group. In some embodiments, the peptide intermediate of compound A42 comprises a glutamic acid residue bonded to the protecting group via a carboxyl group of its side chain. In some embodiments, the glutamic acid residue is covalently bonded to an allyl group. In some embodiments, the peptide intermediate of compound A42 comprises at least one amino acid covalently bonded to a spacer as defined herein. The spacer is further covalently bonded to a protecting group or a hydrogen atom. In some embodiments, the present invention provides peptide intermediates shown in Table 11 above. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 342. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 442. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 242. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 542. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 642. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 742.

[0564] In some embodiments, the present invention relates to a method for preparing compounds of the present invention, such as compound A42 (SEQ ID NO: 42), wherein a polypeptide intermediate such as compound D42 (SEQ ID NO: 342) is activated by the following activated acyl group, namely (LG)CO(CH2), as illustrated in Scheme 2 below. zz The present invention provides a method comprising the step of acylation with CO2H, wherein ZZ is 14-22 and LG is a leaving group as defined herein:

[0565] [Table 12] [Table 13]

[0566] In some embodiments, the present invention provides peptide intermediates of compound A43 (SEQ ID NO: 43). In some embodiments, the peptide intermediate of compound A43 comprises at least one amino acid covalently bonded to a protecting group. In some embodiments, the peptide intermediate of compound A43 comprises a lysine residue bonded to the protecting group via an amino group of its side chain. In some embodiments, the lysine residue is covalently bonded to Alloc or ivDde. In some embodiments, the peptide intermediate of compound A43 comprises an aspartic acid residue bonded to the protecting group via a carboxyl group of its side chain. In some embodiments, the aspartic acid residue is covalently bonded to an allyl group. In some embodiments, the peptide intermediate of compound A43 comprises a glutamic acid residue bonded to the protecting group via a carboxyl group of its side chain. In some embodiments, the glutamic acid residue is covalently bonded to an allyl group. In some embodiments, the peptide intermediate of compound A43 comprises at least one amino acid covalently bonded to a spacer as defined herein. The spacer is further covalently bonded to a protecting group or a hydrogen atom. In some embodiments, the present invention provides peptide intermediates shown in Table 12 above. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 343. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 443. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 243. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 543. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 643. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 743.

[0567] In some embodiments, the present invention relates to a method for preparing compounds of the present invention, such as compound A43 (SEQ ID NO: 43), wherein a polypeptide intermediate such as compound D43 (SEQ ID NO: 343) is activated by the following activated acyl group, namely (LG)CO(CH2), as illustrated in Scheme 3 below. zzThe present invention provides a method comprising the step of acylation with CO2H, wherein ZZ is 14-22 and LG is a leaving group as defined herein:

[0568] [Table 14]

[0569] How to use In another embodiment, the present invention relates to a method for treating metabolic diseases or disorders in subjects requiring treatment, comprising providing an effective amount of the PYY analog polypeptide or a pharmaceutical composition thereof to the subject. Metabolic diseases or disorders include type 1 diabetes, type 2 diabetes, and obesity. Furthermore, the present invention relates to a method for achieving weight loss in subjects including diabetic subjects, comprising providing an effective amount of the PYY analog polypeptide thereof to the subject. In certain embodiments, the present invention also relates to a method for treating non-alcoholic fatty liver disease (NAFLD) and / or non-alcoholic steatohepatitis (NASH).

[0570] In some embodiments, methods are provided for treating obesity in a human subject, causing weight loss in the human subject, or suppressing the appetite of the human subject, the methods comprising administering to the subject any of the polypeptides disclosed herein, a pharmaceutical composition comprising any of the disclosed polypeptides, or an osmotic delivery device comprising any of the disclosed polypeptides.

[0571] In some embodiments, a method is provided for treating diabetes in a human subject, comprising administering to the subject any of the polypeptides disclosed herein, a pharmaceutical composition comprising any of the disclosed polypeptides, or an osmotic delivery device comprising any of the disclosed polypeptides. In some embodiments, the diabetes is type 1 diabetes. In some embodiments, the diabetes is type 2 diabetes. In some embodiments, the present invention provides a method for treating non-alcoholic fatty liver disease (NAFLD) in a human subject, comprising administering to the subject any of the polypeptides disclosed herein, a pharmaceutical composition comprising any of the disclosed polypeptides, or an osmotic delivery device comprising any of the disclosed polypeptides.

[0572] In some embodiments, the present invention provides a method for treating non-alcoholic steatohepatitis (NASH) in a human subject, comprising administering to the subject any of the polypeptides disclosed herein, a pharmaceutical composition comprising any of the disclosed polypeptides, or an osmotic delivery device comprising any of the disclosed polypeptides.

[0573] The PYY analog polypeptides of this disclosure are particularly useful for the treatment of diabetes, and the method comprises providing an effective amount of PYY analog polypeptide to a diabetic subject. In some embodiments, the PYY analog polypeptides of this disclosure are used to treat subjects with type 1 or type 2 diabetes to control or lower the subject's blood glucose levels, where blood glucose levels can be monitored or estimated based on measured blood concentrations of glycated hemoglobin (hemoglobin A1c, HbA1c).

[0574] (i) In some embodiments, the PYY analog polypeptide of the present disclosure is used for the treatment of subjects having type 1 diabetes; (ii) In some embodiments, the PYY analog polypeptide of the present disclosure is used for the treatment of subjects having type 2 diabetes; (iii) In some embodiments, the PYY analog polypeptides of the present disclosure are used for the treatment of obesity; (iv) In some embodiments, the PYY analog polypeptides of the present disclosure are used to reduce weight in subjects such as those with diabetes. (v) In some embodiments, the PYY analog polypeptide of the present disclosure is used for the treatment of non-alcoholic fatty liver disease (NAFLD), (vi) In some embodiments, the PYY analog polypeptide of the present disclosure is used for the treatment of non-alcoholic steatohepatitis (NASH), Here, the PYY analog polypeptide of usage (i), (ii), (iii), (iv), (v) or (vi) includes any isolated polypeptide of the present disclosure, or a pharmaceutically acceptable salt thereof, which includes one represented by any of the consensus sequences from SEQ ID NO: 1 to SEQ ID NO: 78.

[0575] In some embodiments, the PYY analog polypeptide of use (i), (ii), (iii), (iv), (v) or (vi) comprises any isolated polypeptide of the present disclosure, or a pharmaceutically acceptable salt thereof, including those selected from the group consisting of SEQ ID NOs: 13, 24, 42, and 43.

[0576] In some embodiments, the PYY analog polypeptide of usage (i), (ii), (iii), (iv), (v), or (vi) comprises the isolated polypeptide of SEQ ID NO: 13 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide of usage (i), (ii), (iii), (iv), (v), or (vi) comprises the isolated polypeptide of SEQ ID NO: 24 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide of usage (i), (ii), (iii), (iv), (v), or (vi) comprises the isolated polypeptide of SEQ ID NO: 42 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide of usage (i), (ii), (iii), (iv), (v), or (vi) comprises the isolated polypeptide of SEQ ID NO: 43 or a pharmaceutically acceptable salt thereof.

[0577] In some embodiments, the PYY analog polypeptide is used to treat a subject with type 1 diabetes, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 13 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to treat a subject with type 1 diabetes, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 24 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to treat a subject with type 1 diabetes, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 42 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to treat a subject with type 1 diabetes, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 43 or a pharmaceutically acceptable salt thereof.

[0578] In some embodiments, the PYY analog polypeptide is used to treat subjects with type 2 diabetes, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 13 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to treat subjects with type 2 diabetes, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 24 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to treat subjects with type 2 diabetes, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 42 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to treat subjects with type 2 diabetes, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 43 or a pharmaceutically acceptable salt thereof.

[0579] In some embodiments, the PYY analog polypeptide is used to treat obese subjects, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 13 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to treat obese subjects, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 24 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to treat obese subjects, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 42 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to treat obese subjects, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 43 or a pharmaceutically acceptable salt thereof.

[0580] In some embodiments, the PYY analog polypeptide is used to reduce the weight of the subject, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 13 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to reduce the weight of the subject, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 23 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to reduce the weight of the subject, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 42 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to reduce the weight of the subject, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 43 or a pharmaceutically acceptable salt thereof.

[0581] In some embodiments, the PYY analog polypeptide is used to treat a subject having NAFLD, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 13 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to treat a subject having NAFLD, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 24 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to treat a subject having NAFLD, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 42 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to treat a subject having NAFLD, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 43 or a pharmaceutically acceptable salt thereof.

[0582] In some embodiments, the PYY analog polypeptide is used to treat a subject with NASH, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 13 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to treat a subject with NASH, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 24 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to treat a subject with NASH, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 42 or a pharmaceutically acceptable salt thereof. In some embodiments, the PYY analog polypeptide is used to treat a subject with NASH, and the isolated polypeptide is the isolated polypeptide of SEQ ID NO: 43 or a pharmaceutically acceptable salt thereof.

[0583] Certain PYY analog polypeptides of this disclosure, when administered in combination with a GLP-1 receptor agonist, can induce appetite suppression and weight loss effects, while when administered alone, they can induce dose-dependent appetite stimulation and weight gain effects. This property of the selected PYY analog polypeptides of this disclosure is useful for the treatment of various wasting disorders. Therefore, in some embodiments, the PYY analog polypeptides of this disclosure are used for the treatment of conditions such as anorexia nervosa, sarcopenia, frailty, and cachexia. In some embodiments, the PYY analog is SEQ ID NO: 13. In some embodiments, the PYY analog is SEQ ID NO: 24.

[0584] As used herein, the terms “patient” or “subject” refer to rodents or animals, preferably mammals, most preferably humans.

[0585] combination In some embodiments, the PYY analog polypeptide of the Disclosure is combined with a second agent to form a combination. In some embodiments, the PYY analog polypeptide of the Disclosure is combined with a second agent to form a combination, the second agent being an incretin mimetic. In some embodiments, the PYY analog polypeptide of the Disclosure is combined with a second agent to form a combination, the second agent being an insulin-secreting compound.

[0586] As used herein, the phrase "incretin mimetic" means: GLP-1 peptide; GLP-1(7-36); GLP-1 receptor agonist; GLP-1 peptide derivative; GLP-1 peptide analog; exenatide; exenatide having the amino acid sequence of exendin-4 (natural form of exenatide); exenatide-LAR; lixisenatide; liraglutide; semaglutide; dulaglutide; albiglutide; taspoglutide; tylzepatide (Eli This includes, but is not limited to, Lilly's LY3298176 or Y-(Aib)-EGTFTSDYSI-(Aib)-LDKIAQ-[diacid-gamma-Glu-(AEEA)2-Lys]-AFVQWLIAGGPSSGAPPPS-NH2) SEQ ID NO: 805); glucagon and its peptide analogs and peptide derivatives; glucagon-like polypeptide-2 (GLP-2); PYY and its peptide analogs and peptide derivatives; PYY(3-36); oxytomodulin and its peptide analogs and peptide derivatives; amylin and its peptide analogs and peptide derivatives; and gastric suppressor peptide (GIP). Incretin mimetic compounds are also referred to herein as “insulin-secreting peptides.” Incretin mimetic compounds targeting the GLP-1 receptor are also known in the literature as “GLP-1 receptor agonists” or “GLP-1 agonists,” and both terms are used interchangeably herein.

[0587] Some embodiments of the present invention, as non-limiting examples, include the use of the disclosed PYY analog polypeptide in combination with a second therapeutic agent, such as a second polypeptide, such as an insulin-secreting peptide. In some embodiments, a pharmaceutical composition comprising the PYY analog polypeptide in combination with a second agent is used to treat type 2 diabetes.

[0588] In some embodiments, a pharmaceutical composition comprising one of the isolated polypeptides disclosed herein is provided. In some embodiments, a pharmaceutical composition comprising one of the isolated polypeptides disclosed herein, further comprising a second polypeptide, is provided. In some embodiments, the second polypeptide is a glucagon analog. In some embodiments, the second polypeptide is an amyrin analog. In preferred embodiments, the second polypeptide is a GLP-1 receptor agonist.

[0589] The term "GLP-1" refers to a polypeptide, glucagon-like peptide-1(7-36)amide, a 30-residue peptide hormone released from intestinal L cells after nutrient consumption. GLP-1 has the amino acid sequence (HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR-NH2), sequence number 801. GLP-1 is a regulatory peptide that binds to the extracellular domain of the GLP-1 receptor (GLP-1R), a G protein-coupled receptor on β-cells. It stimulates the insulin response to nutrients absorbed from the intestines through adenyl cyclase activity and cAMP production [Baggio 2007, “Biology of incretins: GLP-1 and GIP,” Gastroenterology, vol. 132(6):2131-57; Holst 2008, “The incretin system and its role in type 2 diabetes mellitus,” Mol Cell Endocrinology, vol. 297(1-2):127-36]. The activating mechanisms of GLP-1R are diverse.GLP-1 maintains glucose homeostasis by enhancing endogenous glucose-dependent insulin secretion, increasing glucose sensitivity in β-cells and thus increasing sensitivity to GLP-1, suppressing glucagon release, restoring first and second-stage insulin secretion, delaying gastric emptying, reducing food intake, and increasing satiety. [Holst 2008 Mol. Cell Endocrinology; Kjems 2003 “The influence of GLP-1 on glucose-stimulated insulin secretion: effects on beta-cell sensitivity in type 2 and nondiabetic subjects,” Diabetes, vol. 52(2): 380-86; Holst 2013 “Incretin hormones and the satiation signal,” Int J Obes (Lond), vol. 37(9):1161-69; Seufert 2014, “The extra-pancreatic effects of GLP-1 receptor agonists: a focus on the cardiovascular, [gastrointestinal and central nervous systems, Diabetes Obes Metab, vol. 16(8): 673-88]. Considering the mechanism of action of GLP-1, the risk of hypoglycemia is minimal.

[0590] Glucagon-like peptide-1(7-36)amide (GLP-1) is a 30-residue peptide hormone released from intestinal L cells after nutrient consumption. It enhances glucose-induced insulin secretion from pancreatic beta cells, increases insulin expression, inhibits beta cell apoptosis, promotes beta cell regeneration, reduces glucagon secretion, slows gastric emptying, promotes satiety, and increases peripheral glucose processing. These multiple effects have generated considerable interest in the discovery of long-lasting agonists of the GLP-1 receptor (GLP-1R) for the treatment of type 2 diabetes. As used herein, the term “exenatide” includes, but is not limited to, exenatide, (HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS-NH2), exenatide having the amino acid sequence of SEQ ID NO: 802, natural exenzin-4, exenatide peptide, exenatide peptide analogs, and exenatide peptide derivatives.

[0591] Endogenous GLP-1 is released from the intestines in response to nutrient intake. Following food intake and digestion, carbohydrates and fats appear in the intestinal lumen, stimulating the so-called incretin effect, i.e., the release of incretins such as GLP-1 from intestinal L cells. Once released, GLP-1 targets the pancreas, where it promotes insulin secretion "glucose-dependently." In other words, this GLP-1-mediated effect on insulin persists when blood glucose levels are high, but safely dissipates when blood glucose levels decrease. Therefore, GLP-1 activity autoregulates to reduce the risk of hypoglycemia (a state in which blood glucose levels drop to dangerously low levels). GLP-1 elimination half-life (t 1 / 2 Because its lifespan is short (less than 5 minutes), this endogenous peptide is unsuitable for use as a therapeutic agent due to its short lifespan.

[0592] Synthetic analogs of GLP-1 have a longer half-life and are designed to similarly enhance insulin secretion in a glucose-dependent manner, like endogenous GLP-1, for use in the treatment and weight loss of type 2 diabetes.

[0593] Numerous GLP-1 receptor agonists (e.g., GLP-1 peptide derivatives and peptide analogs) exhibiting insulin secretion-promoting effects have been developed in the art (e.g., U.S. Patent Nos. 5,118,666; 5,120,712; 5,512,549; 5,545,618; 5,574,008; 5,574,008; 5,614,492; 5,958,909; 6,191,102; 6,268,343; 6,329,336; 6,451,974; 6,458,924; 6,514,500; 6,593,29 See No. 5; No. 6,703,359; No. 6,706,689; No. 6,720,407; No. 6,821,949; No. 6,849,708; No. 6,849,714; No. 6,887,470; No. 6,887,849; No. 6,903,186; No. 7,022,674; No. 7,041,646; No. 7,084,243; No. 7,101,843; No. 7,138,486; No. 7,141,547; No. 7,144,863; and No. 7,199,217), and clinical trials (e.g., taspoglutide and albiglutide).

[0594] Certain GLP-1 receptor agonists, including Bydureon® (exenatide), marketed by AstraZeneca of Cambridge, UK; Trulicity® (dulaglutide), marketed by Eli Lilly and Co., of Indianapolis, IN, USA; and Victoza® (liraglutide), Ozempic® (semaglutide for injection), and Rybelsus® (semaglutide for oral administration), marketed by Novo Nordisk A / S of Bagsvaerd, Denmark, have been approved by numerous regulatory authorities, including the U.S. Food and Drug Administration (USFDA) and the European Medicines Agency (EMA), for the treatment of patients with type 2 diabetes. These marketed GLP-1 receptor agonists have been developed and formulated for injectable and / or oral administration to patients. However, patient adherence to injectable and oral GLP-1 receptor agonist treatment for type 2 diabetes is well known to be insufficient, preventing many patients from realizing the potential for complete and sustained treatment with GLP-1 receptor agonists. Many patients skip or discontinue regular self-administration of prescribed injectable and oral GLP-1 receptor agonists, and as a result, are unable to adequately treat and control their type 2 diabetes.

[0595] The PYY analog polypeptide of this disclosure, combined with a GLP-1 receptor agonist, has been found to produce bariatric surgery-like efficacy for weight loss. The observation that improved glycemic control after Roux-en-Y gastric bypass surgery (RYGB) in obese or obese T2D patients precedes weight loss observed after RYGB suggests that surgical reorganization of the intestine leads to physiological adaptations beyond those facilitated solely by weight loss. Indeed, RYGB promotes the postprandial secretion of GLP-1 and PYY, both of which are released from L cells lining the inner surface of the distal intestine. The intestinal peptide hormones GLP-1 and peptide tyrosine-tyrosine (PYY), respectively, are involved in systemic energy balance through several overlapping biological responses to energy input. These responses primarily include enhanced glucose-induced insulin secretion, inhibition of gastric emptying, induction of satiety, and inhibition of food intake.

[0596] Therefore, the combination of the PYY analog polypeptide and the GLP-1 receptor agonist of this disclosure is suitable for the treatment of the diseases and disorders disclosed herein. In some embodiments, the GLP-1 receptor agonist is a long-acting GLP-1 receptor agonist.

[0597] Oxintomodulin is a naturally occurring 37-amino acid peptide hormone found in the colon, known to suppress appetite and promote weight loss (Wynne K, et al., Int J Obes (Lond) 30(12):1729-36 (2006)). The sequences of oxintomodulin, as well as its peptide analogs and derivatives, are known in the art (e.g., Bataille D, et al., Peptides 2(Suppl 2):41-44 (1981); and U.S. Patent Application Publications 2005 / 0070469 and 2006 / 0094652).

[0598] Gastric suppressor peptide (GIP) is an insulin-secreting peptide hormone (Efendic, S., et al., Horm Metab Res. 36:742-6 (2004)) secreted by the mucosa of the duodenum and jejunum in response to absorbed fats and carbohydrates, stimulating the pancreas to secrete insulin. GIP circulates as a biologically active 42-amino acid peptide. GIP is also known as glucose-dependent insulinopromoting protein. GIP is a 42-amino acid gastrointestinal regulatory peptide that stimulates insulin secretion from pancreatic beta cells in the presence of glucose (Tseng, C., et al., PNAS 90:1992-1996 (1993)). The sequence of GIP, as well as its peptide analogs and peptide derivatives, are known in the art (e.g., Meier JJ, Diabetes Metab Res Rev. 21(2):91-117 (2005) and Efendic S., Horm Metab Res. 36(11-12):742-6 (2004)).

[0599] Glucagon is a peptide hormone produced by alpha cells in the pancreas that increases glucose levels in the bloodstream. Its effect is the opposite of insulin, which lowers glucose levels. When blood glucose levels become too low, the pancreas releases glucagon. Glucagon converts glycogen stored in the liver into glucose, which is then released into the bloodstream. High blood glucose levels stimulate insulin release. Insulin enables glucose uptake and use by insulin-dependent tissues. Therefore, glucagon and insulin are part of a feedback system that maintains stable blood glucose levels.

[0600] Human amylin, or islet amyloid polypeptide (IAPP), is a 37-residue polypeptide hormone. Amylin is co-secreted from pancreatic β-cells with insulin in a ratio of approximately 100:1 (insulin:amylin). Pro-islet amyloid polypeptide (i.e., pro-IAPP) is produced in pancreatic β-cells as a 67-amino acid, 7404-Dalton propeptide, which undergoes post-translational modifications, including protease cleavage, to produce 37-residue amylin. Loss of β-cell function, which can occur in the early stages of type 1 diabetes and later in type 2 diabetes, leads to deficiencies in insulin and amylin secretion.

[0601] Amylin functions as part of the pancreatic endocrine tissue, which consists of cells in the pancreas that synthesize and secrete hormones. Amylin contributes to blood glucose control; it is secreted from the pancreatic islets into the bloodstream and cleared by renal peptidases. Amylin's metabolic function is well characterized as an inhibitor of the appearance of nutrients such as glucose in the plasma. Therefore, amylin acts as a synergistic partner of insulin, a peptide that regulates blood glucose levels and modulates the distribution and uptake of glucose in the body. The role of insulin in the body is, among other things, to prevent blood glucose levels from becoming too high, especially after meals.

[0602] Amylin is thought to play a role in blood glucose regulation by slowing gastric emptying and promoting satiety (i.e., bloating), thereby preventing a rapid rise in blood glucose levels after eating. The overall effect is to slow the rate at which blood glucose appears after eating. Amylin also reduces the secretion of glucagon by the pancreas. The role of glucagon in the body is, among other things, to prevent blood glucose levels from becoming too low. This is important, for example, as certain type 1 diabetes patients tend to secrete excessive amounts of glucagon that raise blood glucose levels immediately after meals.

[0603] For several reasons, human amylin, with a half-life of approximately 13 minutes in serum, is not suitable for therapeutic use. Instead, plumrintide (Symlin®, developed by Amylin Pharmaceuticals, Inc., San Diego, CA, USA, and marketed by AstraZeneca plc, Cambridge, UK) was developed as a synthetic analog of human amylin for the treatment of type 1 or type 2 diabetes patients who are using mealtime insulin but are unable to achieve desired glycemic control despite optimal insulin therapy. Plumrintide differs from human amylin in three of its 37 amino acids. These modifications give plumrintide a longer half-life of approximately 48 minutes in humans and a reduced tendency to aggregate, a characteristic of human amylin. Further analogues of human amylin have been disclosed, such as those in U.S. Patent Application No. 16 / 598,915 (corresponding to PCT International Application No. PCT / US2019 / 055696), both filed on October 10, 2019.

[0604] Implantable delivery In some embodiments, an osmotic delivery device as described herein is provided, comprising any of the long-acting PYY analog polypeptides disclosed herein, or a pharmaceutical composition comprising any of the long-acting PYY analog polypeptides.

[0605] In some embodiments, the osmotic delivery device includes: an impermeable reservoir having an inner and outer surface and first and second open ends; a semipermeable membrane in a sealed relationship with the first open end of the reservoir; an osmotic engine located within the reservoir and adjacent to the semipermeable membrane; a piston adjacent to the osmotic engine, the piston forming a movable seal with the inner surface of the reservoir, the piston dividing the reservoir into a first chamber and a second chamber, the first chamber containing the osmotic engine; a suspension formulation, the second chamber containing the suspension formulation, the suspension formulation being fluid and containing isolated polypeptides; and a diffusion moderator inserted into the second open end of the reservoir and adjacent to the suspension formulation.

[0606] An implantable osmotic delivery device typically includes a reservoir having at least one orifice through which a suspension formulation can be delivered. The suspension formulation can be stored in the reservoir. In a preferred embodiment, the implantable drug delivery device is an osmotic delivery device, and drug delivery is driven osmotically. Several penetrating delivery devices and their components, such as the DUROS® delivery vise or similar devices, are described (e.g., U.S. Patent Nos. 5,609,885; 5,728,396; 5,985,305; 5,997,527; 6,113,938; 6,132,420; 6,156,331; 6,217,906; 6,261,584; 6,270,787; 6,287,295; 6,375,978; 6,395,292). See also U.S. Patent Publication Nos. 6,508,808; 6,544,252; 6,635,268; 6,682,522; 6,923,800; 6,939,556; 6,976,981; 6,997,922; 7,014,636; 7,207,982; and 7,112,335; 7,163,688; U.S. Patent Application Publication Nos. 2005 / 0175701, 2007 / 0281024, 2008 / 0091176, and 2009 / 0202608).

[0607] Osmotic delivery devices typically consist of an osmotic engine, a piston, and a cylindrical reservoir containing the drug formulation. The reservoir is covered at one end by a semipermeable membrane with controlled velocity and at the other end by a diffusion moderator through which the drug-containing suspension is released from the drug reservoir. The piston separates the drug formulation from the osmotic engine and, using a seal, prevents water in the osmotic engine section from entering the drug reservoir. The diffusion moderator, in conjunction with the drug formulation, is designed to prevent body fluids from entering the drug reservoir through an orifice.

[0608] Osmotic devices release drugs at a predetermined rate based on the principle of osmosis. Extracellular fluid enters directly into the salt engine of the osmotic delivery device through a semipermeable membrane, and this salt engine expands to drive a piston at a slow and uniform delivery rate. The movement of the piston forces the drug formulation to be released through an orifice or outlet port at a predetermined shear rate. In one embodiment of the present invention, the reservoir of the osmotic device is filled with a suspension formulation, and the device can deliver the suspension formulation to a target at a predetermined, therapeutically effective delivery rate over a long period of time (e.g., about 1, 3, 6, 9, 10, and 12 months).

[0609] The drug release rate from an osmotic delivery device typically provides a target dose of the drug, for example, a therapeutically effective daily dose delivered over a day; that is, the drug release rate from the device provides a substantially steady-state delivery of the drug at a therapeutic concentration to the target.

[0610] Typically, in the case of osmotic delivery devices, the volume of the beneficial drug chamber containing the beneficial drug formulation is between approximately 100 μl and approximately 1000 μl, more preferably between approximately 120 μl and approximately 500 μl, and more preferably between approximately 150 μl and approximately 200 μl.

[0611] Typically, osmotic delivery devices are implanted subcutaneously or subcutaneously within a subject to provide subcutaneous drug delivery. The device(s) can be implanted subcutaneously or subcutaneously in one or both arms (e.g., medial, lateral, or posterior upper arms) or the abdomen. Preferred sites in the abdominal region are subcutaneously in the area extending below the ribs and above the beltline. To provide multiple sites for implanting one or more osmotic delivery devices within the abdomen, the abdominal wall can be divided into four quadrants as follows: the upper right quadrant, extending at least 2-3 centimeters below the right ribs (e.g., at least approximately 5-8 centimeters below the right ribs) and at least 2-3 centimeters to the right of the midline (e.g., at least approximately 5-8 centimeters to the right of the midline); the upper right quadrant, extending at least 2-3 centimeters above the beltline (e.g., at least approximately 5-8 centimeters above the beltline); and the upper right quadrant, extending at least 2-3 centimeters to the right of the midline (e.g., at least approximately 5-8 centimeters to the right of the midline). The area can be divided into: the lower right quadrant, extending approximately 5-8 centimeters; the upper left quadrant, extending at least 2-3 centimeters below the left ribs, for example, at least approximately 5-8 centimeters below the left ribs and at least 2-3 centimeters to the left of the midline, for example, at least approximately 5-8 centimeters to the left of the midline; and the lower left quadrant, extending at least 2-3 centimeters above the beltline, for example, at least approximately 5-8 centimeters above the beltline and at least 2-3 centimeters to the left of the midline, for example, at least approximately 5-8 centimeters to the left of the midline. This provides multiple available sites for implanting one or more devices in one or more occasions. Implantation and removal of osmotic delivery devices are generally performed by medical professionals using local anesthesia (e.g., lidocaine).

[0612] Treatment can be easily terminated by removing the osmotic delivery device from the subject, which offers the significant advantage of immediately stopping drug delivery to the subject.

[0613] Preferably, the osmotic delivery device has a fail-safe mechanism to prevent accidental drug overload or bolus delivery in theoretical situations such as blockage or clogging of the outlet (diffusion moderator) through which the drug formulation is delivered. To prevent accidental drug overload or bolus delivery, the osmotic delivery device is designed and constructed such that the pressure required to partially or completely remove or evacuate the diffusion moderator from the reservoir exceeds the pressure required to partially or completely remove or evacuate the semipermeable membrane to the extent necessary to depressurize the reservoir. In such a scenario, the pressure within the device will build up until the device pushes the semipermeable membrane outward at the other end, thereby releasing the osmotic pressure. The osmotic delivery device then becomes static and no longer delivers the drug formulation if the piston is in a sealed relationship with the reservoir.

[0614] The dose and delivery rate can be selected to achieve the desired blood concentration of the drug within approximately six times the half-life of the drug in the subject, generally after device implantation. The blood concentration of the drug is selected to provide the optimal therapeutic effect of the drug while avoiding undesirable side effects that may be induced by drug overconcentrations, and at the same time avoiding peaks and troughs that may induce side effects associated with the peak or trough plasma concentration of the drug.

[0615] The suspension formulation can also be used with infusion pumps, such as the ALZET® (DURECT Corporation, Cupertino, Calif.) osmotic pump, which is a microinfusion pump for continuous drug administration to laboratory animals (e.g., mice and rats).

[0616] Mode of administration In some embodiments, the method includes providing the PYY analog polypeptide or a pharmaceutical composition thereof to a subject in need of treatment via injection. In some embodiments, the method includes providing the PYY analog polypeptide or a pharmaceutical composition thereof to a subject in need of treatment, formulated for oral administration.

[0617] In some embodiments, the method includes providing the PYY analog polypeptide or a pharmaceutical composition thereof to a subject in need of treatment via implantation. In some embodiments, the method includes providing continuous delivery of the PYY analog polypeptide to a subject in need of treatment from an osmotic delivery device. The delivery device, such as an osmotic delivery device, contains a sufficient amount of the PYY analog polypeptide of the disclosure for continuous administration for up to 3 months, 6 months, 9 months, 12 months, 18 months, or 24 months. Thus, continuous administration of the PYY analog polypeptide of the disclosure via an osmotic delivery device eliminates the need for daily or multiple daily dosing of commercially available PYY analog polypeptides.

[0618] Substantial steady-state delivery of PYY analog polypeptides from osmotic delivery devices is continuous throughout the administration period. In some embodiments, the subject or patient is a human subject or human patient.

[0619] In some embodiments of the present invention, the administration period is, for example, at least about 3 months, at least about 3 months to about 1 year, at least about 4 months to about 1 year, at least about 5 months to about 1 year, at least about 6 months to about 1 year, at least about 8 months to about 1 year, at least about 9 months to about 1 year, at least about 10 months to about 1 year, at least about 1 year to about 2 years, and at least about 2 years to about 3 years.

[0620] In further embodiments, the therapeutic method of the present invention provides a significant reduction in the fasting plasma glucose concentration of a subject after implantation of an osmotic delivery device (compared to the subject's fasting plasma glucose concentration before implantation of the osmotic delivery device), and this reduction is achieved within approximately 7, 6, 5, 4, 3, 2, or 1 day or less after implantation of the osmotic delivery device in the subject. The significant reduction in fasting plasma glucose is typically statistically significant or considered significant for the subject by the physician, as demonstrated by the application of appropriate statistical tests. The significant reduction in fasting plasma glucose compared to baseline before implantation is typically maintained over the duration of administration.

[0621] In some embodiments, the present invention relates to a method for treating a disease or condition in a subject requiring treatment. This method includes providing continuous delivery of a drug from an osmotic delivery device, thereby achieving substantially steady-state delivery of the drug at therapeutic concentrations in the subject. Substantial steady-state delivery of the drug from the osmotic delivery device is continuous over a dosing period of at least about three months. The drug has a known or determined half-life in a typical subject. Humans are a preferred subject for carrying out the present invention. The present invention includes a drug effective in treating a disease or condition, and an osmotic delivery device containing the drug for use in this method for treating a disease or condition in a subject requiring treatment. Advantages of the present invention include reduced peak-related drug toxicity and attenuation of suboptimal drug therapy associated with troughs.

[0622] In some embodiments, substantial steady-state delivery of the drug at therapeutic concentrations is achieved within a period of approximately one month, seven days, five days, three days, or one day after implantation of the osmotic delivery device in the subject.

[0623] The present invention also provides a method for promoting weight loss in subjects requiring weight loss promotion, a method for treating excess weight or obesity in subjects requiring treatment of excess weight or obesity, and / or a method for suppressing appetite in subjects requiring appetite suppression. This method comprises providing delivery of an isolated PYY analog polypeptide. In some embodiments, the isolated PYY analog polypeptide is delivered continuously from an implantable osmotic delivery device. In some embodiments, substantially steady-state delivery of the PYY analog polypeptide from the osmotic delivery device is achieved and is substantially continuous over the duration of administration. In some embodiments, the subject is human.

[0624] The present invention includes an osmotic delivery device containing a PYY analog polypeptide for use in the present method in subjects requiring treatment. Subjects may have type 2 diabetes. Subjects requiring treatment may have a baseline HbA1c% greater than 10.0%, i.e., may be high baseline (HBL) subjects. Subjects may not have previously received medication for the treatment of type 2 diabetes.

[0625] In further embodiments, the therapeutic method of the present invention results in a significant decrease in the fasting plasma glucose concentration of a subject after implantation of an osmotic delivery device (compared to the subject's fasting plasma glucose concentration before implantation of the osmotic delivery device), and this significant decrease is achieved within approximately 7 days or less after implantation of the osmotic delivery device in the subject, within approximately 6 days or less after implantation of the osmotic delivery device in the subject, within approximately 5 days or less after implantation of the osmotic delivery device in the subject, within approximately 4 days or less after implantation of the osmotic delivery device in the subject, within approximately 3 days or less after implantation of the osmotic delivery device in the subject, within approximately 2 days or less after implantation of the osmotic delivery device in the subject, or within approximately 1 day or less after implantation of the osmotic delivery device in the subject. In preferred embodiments of the present invention, the significant decrease in the fasting plasma glucose concentration of a subject after implantation of an osmotic delivery device, compared to the subject's fasting plasma glucose concentration before implantation, is achieved within approximately 2 days or less after implantation of the osmotic delivery device in the subject, preferably within approximately 1 day or less, or more preferably within approximately 1 day after implantation of the osmotic delivery device in the subject. A significant decrease in fasting plasma glucose is typically statistically significant or considered significant to the subject by the physician, as demonstrated by the application of appropriate statistical tests. A significant decrease in fasting plasma glucose compared to baseline before implantation is typically maintained over the duration of administration.

[0626] In all embodiments of the present invention relating to a method for treating a disease or condition in a subject, an exemplary osmotic delivery device includes, namely, an impermeable reservoir including an inner and outer surface and first and second open ends; a semipermeable membrane in a sealed relationship with the first open end of the reservoir; an osmotic engine located within the reservoir and adjacent to the semipermeable membrane; a piston adjacent to the osmotic engine, the piston forming a movable seal with the inner surface of the reservoir, the piston dividing the reservoir into a first chamber and a second chamber, the first chamber containing the osmotic engine; a drug formulation or a suspension formulation containing a drug, the second chamber containing the drug formulation or suspension formulation, and the drug formulation or suspension formulation being fluid; and a diffusion moderator inserted into the second open end of the reservoir and adjacent to the suspension formulation. In preferred embodiments, the reservoir is made of titanium or a titanium alloy.

[0627] In all embodiments of the present invention relating to a method for treating a target disease or condition, the drug formulation may include a drug and a vehicle formulation. Alternatively, a suspension formulation may be used in the method and may include, for example, a particulate formulation containing a drug and a vehicle formulation. The vehicle formulation used in forming the suspension formulation of the present invention may include, for example, a solvent and a polymer.

[0628] The reservoir of an osmotic delivery device may contain, for example, titanium or a titanium alloy.

[0629] In all embodiments of the present invention, subcutaneous delivery can be provided using an implanted osmotic delivery device.

[0630] In all embodiments of the present invention, continuous delivery may be, for example, zero-order controlled continuous delivery.

[0631] Pharmaceutical composition According to another embodiment, the present invention provides a pharmaceutical composition comprising the compound of the present invention, namely an isolated polypeptide or a pharmaceutically acceptable derivative thereof, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

[0632] In some embodiments, a pharmaceutical composition is provided comprising any of the disclosed polypeptides formulated as a pharmaceutically acceptable salt thereof, such as trifluoroacetate, acetate, or hydrochloride. In some embodiments, a pharmaceutical composition is provided comprising any of the disclosed polypeptides formulated as trifluoroacetate. In some embodiments, a pharmaceutical composition is provided comprising any of the disclosed polypeptides formulated as acetate. In some embodiments, a pharmaceutical composition is provided comprising any of the disclosed polypeptides formulated as hydrochloride.

[0633] The term "pharmaceutically acceptable carrier, adjuvant, or vehicle" refers to a non-toxic carrier, adjuvant, or vehicle that does not impair the pharmacological activity of the compound being formulated.

[0634] As used herein, the term “pharmaceutically acceptable carrier” is intended to include all solvents, polymers, dispersions, coatings, antimicrobial and antifungal agents, isotonic agents and absorption retarders, etc., that are compatible with pharmaceutical administration. Suitable carriers are listed in the latest edition of Remington's Pharmaceutical Sciences, the standard reference text in the art, which is incorporated herein by reference. Preferred examples of such carriers or diluents include, but are not limited to, water, physiological saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Non-aqueous media such as liposomes and non-volatile oils may also be used. The use of such media and agents for pharmaceutically active substances is well known in the art. Unless any conventional media or agent is incompatible with the active compound, its use in the composition is intended. Complementary active ingredients may also be incorporated into the composition.

[0635] Representative pharmaceutically acceptable carriers, adjuvants, or vehicles that may be used in the compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphates, saturated vegetable fatty acids such as glycine, sorbic acid, potassium sorbate, and protamine sulfate, water, salts, or partial glyceride mixtures of electrolytes, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and lanolin.

[0636] In some embodiments, a pharmaceutical composition is provided comprising a pharmaceutically acceptable derivative of any of the disclosed polypeptides, formulated as a pharmaceutically acceptable salt thereof, such as a trifluoroacetate, acetate, or hydrochloride. “pharmaceutically acceptable derivative” means any non-toxic salt, ester, salt of an ester, or other derivative of the compound of the present invention that can directly or indirectly provide the compound of the present invention, or its active metabolite or residue, to a recipient upon administration.

[0637] A pharmaceutical composition contains a drug and can be formulated as a “particulate formulation,” as described in more detail below. The pharmaceutical composition and / or particulate formulation may contain stabilizing components (also referred to herein as “excipients”). Examples of stabilizing components include, but are not limited to, carbohydrates, antioxidants, amino acids, buffers, inorganic compounds, and surfactants.

[0638] The amount of the compound in the composition of the present invention is such that it is effective in measurably activating one or more PYY receptors in a biological sample or patient (e.g., human, rat, monkey, etc.). In certain embodiments, the amount of the compound in the composition of the present invention is such that it is effective in measurably activating human PYY receptors in a biological sample or patient, in the absence or presence of human serum albumin. In certain embodiments, the composition of the present invention is formulated for administration to a patient requiring such a composition. In some embodiments, the composition of the present invention is formulated for injectable administration to a patient. In some embodiments, the composition of the present invention is formulated for administration to a patient via an implantable delivery device, such as an osmotic delivery device.

[0639] The isolated polypeptides of this disclosure (also referred to herein as “active compounds”), and their derivatives, fragments, analogs, and homologs can be incorporated into pharmaceutical compositions suitable for administration. Such compositions typically comprise the isolated polypeptide or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0640] The pharmaceutical compositions of the present invention are formulated to suit their intended route of administration. Examples of routes of administration include parenteral (e.g., intravenous, intradermal, subdermal, subcutaneous), oral (e.g., inhalation), transdermal (i.e., topical), transmucosal, rectal, or combinations thereof. In some embodiments, the pharmaceutical compositions or isolated polypeptides of the present disclosure are formulated for administration by topical administration. In some embodiments, the pharmaceutical compositions or isolated polypeptides of the present disclosure are formulated for administration by inhalation administration. In some embodiments, the pharmaceutical compositions are suitable for subdermal or subcutaneous implantation and are formulated for administration by a device or other suitable delivery mechanism for subcutaneous delivery of the pharmaceutical composition. In some embodiments, the pharmaceutical compositions are suitable for subdermal or subcutaneous implantation and are formulated for administration by an implantable device for subcutaneous delivery of the pharmaceutical composition. In some embodiments, the pharmaceutical compositions are suitable for subdermal or subcutaneous implantation and are formulated for administration by an osmotic delivery device, such as an implantable osmotic delivery device, for subcutaneous delivery of the pharmaceutical composition. Solutions or suspensions used for parenteral, intradermal, subdermal, or combination thereof may include the following components: sterile diluents such as water for injection, physiological saline, non-volatile oils, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; antimicrobial agents such as benzyl alcohol or methylparaben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates, or phosphates; and isotonic modifiers such as sodium chloride or dextrose. pH can be adjusted with an acid or base such as hydrochloric acid or sodium hydroxide. Parenteral preparations may be sealed in glass or plastic ampoules, disposable syringes, or multi-dose vials.

[0641] Suitable pharmaceutical compositions for injection include sterile aqueous solutions (if water-soluble) or dispersions and sterile powders for preparing sterile injection solutions or dispersions immediately before use. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL ((BASF, Parsippany, NJ)), or phosphate-buffered saline (PBS). In all cases, the composition must be sterile and fluid enough to be easily injected. The composition must be stable under manufacturing and storage conditions and protected from contamination by microorganisms such as bacteria or fungi. The carrier may 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. Appropriate fluidity can be achieved, for example, by lecithin. The required particle size can be maintained by the use of any coating, in the case of dispersion, and by the use of surfactants. Prevention of microbial action can be achieved by various antimicrobial and antifungal agents, such as parabens, chlorobutanol, phenol, ascorbic acid, and thimerosal. In many cases, it is preferable to include isotonic agents, such as sugar and polyalcohols (mannitol, sorbitol, sodium chloride, etc.) in the composition. Long-term absorption of the injectable composition can be achieved by including absorption-delaying agents, such as aluminum monostearate and gelatin, in the composition.

[0642] Sterile injectable solutions can be prepared by incorporating the required amount of the active compound, along with one or a combination of the components listed above as needed, into a suitable solvent, followed by sterilization by filtration. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle containing a basic dispersion medium and other required components from those listed above. In the case of sterile powders for the preparation of sterile injectable solutions, the preparation methods are vacuum drying and freeze-drying, which yield a powder with the active ingredient and any additional desired components added from its pre-sterilized filtered solution.

[0643] Oral compositions generally contain an inert diluent or food carrier. They can be encapsulated in gelatin capsules or compressed into tablets. For oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, lozenges, or capsules. Oral compositions can also be prepared using a fluid carrier for use as an oral rinse, in which the compound in the fluid carrier is applied orally, swished in the mouth, and then spat out or swallowed. Pharmaceutically compatible binders and / or adjuvant materials may be included as part of the composition. Tablets, pills, capsules, lozenges, etc., may contain any of the following ingredients or compounds of similar properties: binders such as microcrystalline cellulose, tragacanth gum, or gelatin; excipients such as starch or lactose; disintegrants such as alginic acid, Primogel, or corn starch; lubricants such as magnesium stearate or Sterotes; flow enhancers such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; or flavorings such as peppermint, methyl salicylate, or orange flavor.

[0644] When administered by inhalation, the compound is delivered in the form of an aerosol spray from a pressurized container or dispenser containing a suitable propellant, such as a gas like carbon dioxide, or a nebulizer.

[0645] Systemic administration may also be by mucosal or percutaneous means. For mucosal or percutaneous administration, a penetrating agent suitable for the barrier to which penetration is to occur is used in the formulation. Such penetrating agents are commonly known in the art and include, for example, surfactants, bile salts, and fusidic acid derivatives for mucosal administration. Mucosal administration can be carried out using nasal sprays or suppositories. For percutaneous administration, the active compound is formulated into an ointment, plaster, gel, or cream, as is commonly known in the art.

[0646] In one embodiment, the active compound may be prepared with a carrier that protects the compound from rapid elimination from the body, such as a controlled-release formulation comprising an implant and a microencapsulation delivery system. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydride, polyglycolic acid, collagen, polyorthoester, and polylactic acid can be used. Methods for preparing such formulations will be apparent to those skilled in the art. The materials are also commercially available from Alza Corporation and Nova Pharmaceuticals, Inc. 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.

[0647] For ease of administration and uniformity of dosage, it is particularly advantageous to formulate oral or parenteral compositions in dosage unit form. As used herein, a dosage unit form refers to a physically distinct unit suitable as a unit dose for treating a subject, each unit containing a predetermined amount of the active compound, calculated to produce the desired therapeutic effect, along with the required pharmaceutical carrier. The specifications of the dosage unit forms of the present invention are determined by and directly depend on the inherent characteristics of the active compound, the specific therapeutic effect to be achieved, and the essential limitations of the art in formulating such active compounds for the treatment of an individual.

[0648] The pharmaceutical composition may be contained within a container, pack, or dispenser, along with instructions for administration.

[0649] Drug particle formulations A compound for use in carrying out the present invention, namely an isolated polypeptide or a pharmaceutically acceptable salt thereof, is typically added to a particle formulation to form a suspension formulation by creating polypeptide-containing particles that are uniformly suspended, dissolved, or dispersed in a suspension vehicle. In some embodiments, the PYY analog polypeptide is formulated into a particle formulation and converted into particles (e.g., by spray drying). In some embodiments, particles containing the PYY analog polypeptide are suspended in a vehicle formulation, resulting in a suspension formulation of suspended particles containing the PYY analog polypeptide.

[0650] Preferably, the particle formulation can be formed into particles using processes such as spray drying, freeze-drying, drying, freeze-drying, grinding, granulation, ultrasonic droplet generation, crystallization, precipitation, or other techniques available in the art for forming particles from a mixture of components. In one embodiment of the present invention, the particles are spray-dried. The particles are preferably substantially uniform in shape and particle size.

[0651] In some embodiments, the present invention provides drug particle formulations for pharmaceutical use. The particle formulations typically contain a drug and include one or more stabilizing components (also referred to herein as “excipients”). Examples of stabilizing components include, but are not limited to, carbohydrates, antioxidants, amino acids, buffers, inorganic compounds, and surfactants. The amount of stabilizer in the particle formulation can be determined experimentally based on the activity of the stabilizer and the desired properties of the formulation, taking into account the teachings herein.

[0652] In any embodiment, the particulate formulation may contain about 50% to about 90% by weight of the drug, about 50% to about 85% by weight of the drug, about 55% to about 90% by weight of the drug, about 60% to about 90% by weight of the drug, about 65% to about 85% by weight of the drug, about 65% to about 90% by weight of the drug, about 70% to about 90% by weight of the drug, about 70% to about 85% by weight of the drug, about 70% to about 80% by weight of the drug, or about 70% to about 75% by weight of the drug.

[0653] Typically, the amount of carbohydrates in a particulate formulation is determined by concerns about aggregation. In general, the amount of carbohydrates should not be too high to avoid promoting crystal growth in the presence of water due to excess carbohydrates that are not bound to the drug.

[0654] Typically, the amount of antioxidants in a particle formulation is determined by the risk of oxidation, while the amount of amino acids in the formulation is determined by the risk of oxidation and / or the moldability of the particles during spray drying.

[0655] Typically, the amount of buffer in a particle formulation is determined by pretreatment concerns, stability concerns, and the moldability of the particles during spray drying. If all stabilizers are solubilized, buffer may be needed to stabilize the drug during processes such as solution preparation and spray drying.

[0656] Examples of carbohydrates that may be included in particulate formulations include, but are not limited to, monosaccharides (e.g., fructose, maltose, galactose, glucose, D-mannose, and sorbose), disaccharides (e.g., lactose, sucrose, trehalose, and cellobiose), polysaccharides (e.g., raffinose, melegitose, maltodextrin, dextran, and starch), and algitols (acyclic polyols; e.g., mannitol, xylitol, maltitol, lactitol, xylitol, sorbitol, pyranosylsorbitol, and myoinsitol). Preferred carbohydrates include disaccharides and / or non-reducing sugars, such as sucrose, trehalose, and raffinose.

[0657] Examples of antioxidants that can be included in particulate formulations include, but are not limited to, methionine, ascorbic acid, sodium thiosulfate, catalase, platinum, ethylenediaminetetraacetic acid (EDTA), citric acid, cysteine, thioglycerol, thioglycolic acid, thiosorbitol, butylated hydroxyanisole, butylated hydroxytoluene, and propyl gallate. Furthermore, readily oxidizable amino acids, such as cysteine, methionine, and tryptophan, can be used as antioxidants.

[0658] Examples of amino acids that can be included in particle formulations include, but are not limited to, arginine, methionine, glycine, histidine, alanine, leucine, glutamic acid, isoleucine, L-threonine, 2-phenylamine, valine, norvaline, proline, phenylalanine, tryptophan, serine, asparagine, cysteine, tyrosine, lysine, and norleucine. Preferred amino acids include those that are easily oxidized, such as cysteine, methionine, and tryptophan.

[0659] Examples of buffers that may be included in particulate formulations include, but are not limited to, citrate, histidine, succinate, phosphate, maleate, Tris, acetate, carbohydrates, and glycylglycine. Preferred buffers include citrate, histidine, succinate, and Tris.

[0660] Examples of inorganic compounds that may be included in particulate formulations include, but are not limited to, NaCl, Na2SO4, NaHCO3, KCl, KH2PO4, CaCl2, and MgCl2.

[0661] Furthermore, particulate formulations may contain surfactants and other stabilizers / excipients such as salts. Examples of surfactants include, but are not limited to, polysorbate 20, polysorbate 80, PLURONIC® (BASF Corporation, Mount Olive, NJ) F68, and sodium dodecyl sulfate (SDS). Examples of salts include, but are not limited to, sodium chloride, calcium chloride, and magnesium chloride.

[0662] The particles are typically sized to be delivered via an implantable osmotic delivery device. Uniform particle shape and size typically help provide a consistent and uniform release rate from such a delivery device; however, particle preparations with non-normal particle size distribution profiles can also be used. For example, in a typical implantable osmotic delivery device with a delivery orifice, the particle size is less than about 30%, more preferably less than about 20%, and more preferably less than 10% of the diameter of the delivery orifice. In one embodiment of a particle formulation for use with an osmotic delivery system where the delivery orifice diameter of the implant is about 0.5 mm, the particle size may be, for example, less than about 150 microns to about 50 microns. In one embodiment of a particle formulation for use with an osmotic delivery system where the delivery orifice diameter of the implant is about 0.1 mm, the particle size may be, for example, less than about 30 microns to about 10 microns. In one embodiment, the orifice is about 0.25 mm (250 microns), and the particle size is about 2 microns to about 5 microns.

[0663] Those skilled in the art will understand that a collection of particles follows the principle of particle size distribution. Methods widely used to describe particle size distribution, and recognized in the art, include, for example, the average diameter and the D-value, such as the D50 value, which is commonly used to represent the average diameter of a range of particle sizes in a given sample.

[0664] The particles in the particulate formulation have a diameter between approximately 2 microns and approximately 150 microns, for example, less than 150 microns, less than 100 microns, less than 50 microns, less than 30 microns, less than 10 microns, less than 5 microns, and about 2 microns in diameter. Preferably, the particles have a diameter between approximately 2 microns and approximately 50 microns.

[0665] The particles in a particle formulation containing isolated PYY analog polypeptide have an average diameter between approximately 0.3 microns and approximately 150 microns. The particles in a particle formulation containing isolated PYY analog polypeptide have an average diameter between approximately 2 microns and approximately 150 microns, for example, an average diameter of less than 150 microns, an average diameter of less than 100 microns, an average diameter of less than 50 microns, an average diameter of less than 30 microns, an average diameter of less than 10 microns, an average diameter of less than 5 microns, and an average diameter of approximately 2 microns. In some embodiments, the particles have an average diameter between approximately 0.3 microns and 50 microns, for example, between approximately 2 microns and approximately 50 microns. In some embodiments, the particles have an average diameter between 0.3 microns and 50 microns, for example, between approximately 2 microns and approximately 50 microns, and each particle has a diameter of less than approximately 50 microns.

[0666] Typically, when incorporated into a suspension vehicle, the particles of a particulate formulation do not settle for less than about 3 months at the delivery temperature, preferably less than about 6 months, more preferably less than about 12 months, more preferably less than about 24 months at the delivery temperature, and most preferably less than about 36 months at the delivery temperature. The suspension vehicle typically has a viscosity between about 5,000 and about 30,000 poise, preferably between about 8,000 and about 25,000 poise, and more preferably between about 10,000 and about 20,000 poise. In one embodiment, the suspension vehicle has a viscosity of about 15,000 poise plus or minus about 3,000 poise. Generally speaking, smaller particles tend to settle more slowly in viscous suspension vehicles than larger particles. Therefore, micron to nano-sized particles are typically desirable. In viscous suspension formulations, the particles of the present invention, measuring approximately 2 to 7 microns, will not settle at room temperature for at least 20 years, based on simulation modeling studies. One embodiment of the particle formulation of the present invention includes particles with a size in the range of less than approximately 50 microns, more preferably less than approximately 10 microns, and more preferably between approximately 2 and 7 microns, for use in implantable osmotic delivery devices.

[0667] In summary, the disclosed polypeptide or a pharmaceutically acceptable salt thereof is formulated into a dry powder of solid particles, which maintains the maximum chemical and biological stability of the drug. The particles provide long-term storage stability at high temperatures, thus enabling the delivery of a stable and bioeffective drug to the subject over extended periods. The particles are administered to the patient suspended in a suspension vehicle.

[0668] Particle suspension in vehicle In one embodiment, the suspension vehicle provides a stable environment in which the drug particle formulation is dispersed. The drug particle formulation is chemically and physically stable in the suspension vehicle (as described above). The suspension vehicle typically comprises one or more polymers and one or more solvents that form a solution of sufficient viscosity to uniformly suspend the drug-containing particles. The suspension vehicle may also comprise further components, including but not limited to surfactants, antioxidants, and / or other compounds soluble in the vehicle.

[0669] The viscosity of the suspension vehicle is typically sufficient to prevent the drug particle formulation from settling during storage and to be suitable for use in delivery methods, such as implantable osmotic delivery devices. The suspension vehicle is biodegradable in that it disintegrates or decomposes over a period of time in response to the biological environment, while the drug particles dissolve in the biological environment, and the active pharmaceutical ingredient (i.e., the drug) in the particles is absorbed.

[0670] In the embodiment, the suspension vehicle is a “single-phase” suspension vehicle, which is a homogeneous system of solid, semi-solid, or liquid that is physically and chemically uniform throughout.

[0671] The solvent in which the polymer is dissolved can affect the properties of the suspension formulation, such as the behavior of the drug particle formulation during storage. The solvent can be selected in combination with the polymer such that the resulting suspension vehicle exhibits phase separation when it comes into contact with an aqueous environment. In some embodiments of the present invention, the solvent can be selected in combination with the polymer such that the resulting suspension vehicle exhibits phase separation when it comes into contact with an aqueous environment containing less than approximately 10% water.

[0672] The solvent may be an acceptable solvent that is immiscible with water. The solvent may also be selected so that the polymer is soluble in the solvent at high concentrations, such as a polymer concentration of about 30% or higher. Examples of solvents useful for carrying out the present invention include, but are not limited to, lauryl alcohol, benzyl benzoate, benzyl alcohol, lauryl lactate, decanol (also called decyl alcohol), ethylhexyl lactate, and long-chain (C8-C24) aliphatic alcohols, esters, or mixtures thereof. The solvent used in the suspension vehicle may be "dry" in that it has a low water content. Preferred solvents for use in the formulation of the suspension vehicle include lauryl lactate, lauryl alcohol, benzyl benzoate, and mixtures thereof.

[0673] Examples of polymers for the formulation of the suspension vehicle of the present invention include, but are not limited to, polyesters (e.g., polylactic acid and polyglycolic acid polylactic acid), polymers containing pyrrolidone (e.g., polyvinylpyrrolidone having a molecular weight in the range of approximately 2,000 to approximately 1,000,000), esters or ethers of unsaturated alcohols (e.g., vinyl acetate), polyoxyethylene polyoxypropylene block copolymers, or mixtures thereof. Polyvinylpyrrolidone can be characterized by its K value (e.g., K-17), which is a viscosity index. In one embodiment, the polymer is polyvinylpyrrolidone with a molecular weight of 2,000 to 1,000,000. In a preferred embodiment, the polymer is polyvinylpyrrolidone K-17 (typically having an approximate average molecular weight range of approximately 7,900 to 10,800). The polymer used in the suspension vehicle may comprise one or more different polymers, or a single polymer of different grades. The polymer used in the suspension vehicle may also be dry or have a low water content.

[0674] Generally speaking, the composition of the suspension vehicle for use in the present invention may vary based on the desired performance characteristics. In one embodiment, the suspension vehicle may contain about 40% to about 80% by weight of polymer(s) and about 20% to about 60% by weight of solvent(s). Preferred embodiments of the suspension vehicle include a vehicle formed from polymer(s) and solvent(s) mixed in the following ratios: about 25% by weight of solvent and about 75% by weight of polymer; about 50% by weight of solvent and about 50% by weight of polymer; or about 75% by weight of solvent and about 25% by weight of polymer. Thus, in some embodiments, the suspension vehicle may contain selected components, and in other embodiments, it may essentially consist of selected components.

[0675] Suspension vehicles may exhibit Newtonian behavior. Typically, suspension vehicles are formulated to provide a viscosity that maintains the uniform dispersion of particulate formulations for a given period. This helps facilitate the manufacture of suspension formulations tailored to provide controlled delivery of the drug contained in the drug particulate formulation. The viscosity of a suspension vehicle may vary depending on the desired application, the size and type of the particulate formulation, and the amount of particulate formulation packed into the suspension vehicle. The viscosity of a suspension vehicle may also be altered by changing the type or relative amount of solvent or polymer used.

[0676] The suspension vehicle may have a viscosity in the range of about 100 poise to about 1,000,000 poise, preferably about 1,000 poise to about 100,000 poise. In a preferred embodiment, the suspension vehicle typically has a viscosity between about 5,000 to about 30,000 poise, preferably between about 8,000 to about 25,000 poise, and more preferably between about 10,000 to about 20,000 poise at 33°C. In one embodiment, the suspension vehicle has a viscosity of about 15,000 poise plus or minus about 3,000 poise at 33°C. Viscosity can be measured using a parallel-plate rheometer at 33°C with a shear rate of 10⁻⁴ / sec. Suspension vehicles may exhibit phase separation upon contact with an aqueous environment; however, typically, suspension vehicles do not exhibit substantial phase separation depending on temperature. For example, at temperatures in the range of approximately 0°C to approximately 70°C, and during temperature cycles such as 4°C to 37°C to 4°C, suspension vehicles typically do not exhibit phase separation.

[0677] The suspension vehicle can be prepared by combining a polymer and a solvent under dry conditions, such as in a dry box. The polymer and solvent can be combined and liquefied to form a single phase by raising the temperature, such as to approximately 40°C to 70°C. The components may be blended under reduced pressure to remove bubbles generated from the dry components. The components can be combined using a conventional mixer, such as a double-helix blade or similar mixer, set to a speed of approximately 40 rpm. However, the components can also be mixed using a higher speed. Once a liquid solution of the components is obtained, the suspension vehicle can be cooled to room temperature. Differential scanning calorimetry (DSC) can be used to confirm that the suspension vehicle is single-phase. Furthermore, the components of the vehicle (e.g., solvent and / or polymer) may be treated to substantially reduce or remove peroxides (e.g., by treatment with methionine; see, for example, U.S. Patent Application Publication 2007-0027105).

[0678] A drug particle formulation is added to a suspension vehicle to form a suspension formulation. In some embodiments, the suspension formulation may comprise the drug particle formulation and the suspension vehicle, while in other embodiments, it essentially consists of the drug particle formulation and the suspension vehicle.

[0679] A suspension formulation can be prepared by dispersing a particulate formulation in a suspension vehicle. The suspension vehicle may be heated, and the particulate formulation may be added to the suspension vehicle under dry conditions. The components can be mixed under reduced pressure at a rising temperature, such as about 40°C to about 70°C. The components can be mixed at a sufficient speed, such as about 40 rpm to about 120 rpm, and for a sufficient time, such as about 15 minutes, to achieve uniform dispersion of the particulate formulation in the suspension vehicle. The mixer may be a double-helix blade mixer or another suitable mixer. The resulting mixture can be removed from the mixer, sealed in a dry container to prevent water from contaminating the suspension formulation, and further cooled to room temperature before use, for example, before filling into implantable drug delivery devices, unit dose containers, or multi-dose containers.

[0680] The suspension formulation typically has a total water content of less than about 10% by weight, preferably less than about 5% by weight, and more preferably less than 4% by weight.

[0681] In preferred embodiments, the suspension formulation of the present invention is substantially homogeneous and fluid, providing delivery of a drug particle formulation from an osmotic delivery device to a target.

[0682] In summary, the components of a suspension vehicle provide biocompatibility. The components of a suspension vehicle provide suitable chemical and physical properties for forming a stable suspension of drug particle formulations. These properties include, but are not limited to, the viscosity of the suspension; the purity of the vehicle; the residual moisture content of the vehicle; the density of the vehicle; compatibility with dry powders; compatibility with implantable devices; the molecular weight of the polymer; the stability of the vehicle; and the hydrophobicity and hydrophilicity of the vehicle. These properties can be manipulated and controlled, for example, by manipulating the vehicle composition and the ratios of the components used in the suspension vehicle.

[0683] The suspension formulations described herein can be used in an implantable osmotic delivery device to provide zero-order, continuous, controlled, and sustained delivery of compounds over long periods, such as several weeks, several months, or up to approximately one year or more. Such an implantable osmotic delivery device can typically deliver a drug-containing suspension formulation at a desired flow rate over a desired period. The suspension formulation can be loaded into the implantable osmotic delivery device by conventional techniques. [Examples]

[0684] The following examples are provided to those skilled in the art to provide a complete disclosure and explanation of how to put the invention into practice, and are not intended to limit the scope of what the inventors consider to be the invention. While efforts have been made to ensure accuracy to the numerical values ​​used (e.g., quantities, concentrations, and percentage changes), some experimental errors and deviations should be taken into account. Unless otherwise specified, temperatures are in Celsius, and atmospheric pressure is atmospheric pressure or near atmospheric pressure.

[0685] Example 1: Production of long-acting PYY analog polypeptides The long-acting PYY analog polypeptides of the present invention, provided in Table 3, are synthesized using the Prelude peptide synthesizer (Protein Technologies). At Tucson, AZ, Inc., Rink Amide MBHA LL was synthesized by a solid-phase method using the Fmoc strategy with activation (5-fold molar excess) of N-[((dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridine-1-ylmethylene]-N-methylmethaneaminium hexafluorophosphate N-oxide (HATU) or 2-(6-chloro-1-H-benzotriazol-1-yl)-1,1,3,3-tetramethylaminium hexafluorophosphate (HCTU) in N,N-dimethylformamide (DMF), with N'N-diisopropylethylamine (DIEA) used as the base. A 20% piperidine / DMF solution was used for Fmoc deprotection. The resin used was Rink Amide MBHA LL (Novabiochem) with a loading of (0.30-0.40) mmol / g on a (20-400) μmol scale.

[0686] After the solid-phase synthesis of the linear polypeptide was completed, the resin was washed with dichloromethane (DCM) and dried under vacuum for 30 minutes. For analogues containing an allyloxycarbonyl (Alloc) protecting group, its removal was carried out via a solution of Pd(PPh3)3 in (chloroform / acetic acid / n-methylmorpholine, 37:2:1). For analogues containing tert-butyloxycarbonyl (BOC)-Lys-fluorenylmethyloxycarbonyl (Fmoc)-OH, the Fmoc protecting group was removed using 20% ​​piperidine / DMF. The resulting Fmoc-deprotected resin was washed with DMF (6 × 30 seconds). Next, the extension of the spacer region was carried out stepwise by manually adding each building block under pre-activation conditions. The addition of lipophilic substituents (also called "acyl chains") was carried out under solid-phase peptide synthesis (SPPS) conditions without a pre-activation step. The final deprotection and cleavage of the peptides from the solid support was performed by treating the resin with (95% TFA, 2% water, 2% thioanisole, and 1% triisopropylsilane) for 2-3 hours. The cleaved peptides were precipitated using cold diethyl ether. The diethyl ether layer was decanted, the solid was again pulverized with cold diethyl ether, and pelletized by centrifugation.

[0687] For analogues containing lactam crosslinks, appropriate allyl-protected amino acid building blocks were set up under the normal solid-phase conditions described above. Fmoc-Lys-ε-1-(4,4-dimethyl-2,6-dioxocyclohexa-1-ylidene)-3-methylbutyl(ivDde)-OH was also set up as a handle, and the acyl spacer and side chain were later incorporated. Once the linear peptide was complete, the allyl protecting group was removed as described above. Lactam crosslink formation was achieved by a solid-phase protocol using benzotriazole-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP, 0.5M) activation and DIEA as the base. Deprotection of the Fmoc and ivDde groups was achieved via a 4% solution of hydrazine in DMF. The resulting Fmoc-deprotected resin was washed with DMF (6 × 30 sec). Extension of the spacer region and addition of lipophilic substituents were carried out as described in the previous paragraph. The final deprotection and cleavage of the peptides from the solid support was performed by treating the resin with (95% TFA, 2% water, 2% thioanisole, and 1% triisopropylsilane) for 2-3 hours. The cleaved peptides were precipitated using cold diethyl ether. The diethyl ether was decanted, the solid was again pulverized with cold diethyl ether, and pelletized by centrifugation.

[0688] Next, the crude product was dissolved in a solution of acetonitrile (ACN) / H2O and 0.1% TFA. A 10% acetic acid solution was added to each solution of the crude peptide product, and the mixture was stirred until LC / MS analysis showed removal of the CO2 adduct. The solutions were frozen and lyophilized. Purification was carried out by the method described in Example 2.

[0689] Example 2: Purification and characterization of long-acting PYY receptor agonist polypeptides, i.e., linear polypeptides, that do not contain lipophilic substituents and optional spacers. The product from Example 1 was freeze-dried and analyzed by electrospray ionization-liquid chromatography / mass spectrometry (ESI-LC / MS) and high-pressure liquid chromatography (HPLC) to demonstrate its purity (>98%). The mass results were in agreement with the calculated values.

[0690] Peptide analogs were characterized using either Method A, Method B, Method C, or Method D, by C18 HPLC and LC / MS analysis (Acquity SQD Waters Corp, Milford, MA), as well as UV detection obtained by dual absorbance signals at 215 nm and 280 nm.

[0691] Method A, LC / MS conditions: LC / MS was performed for 30 minutes using a Phenomenex HPLC Aeris® Peptide XB Cl8 35 column, 1.7 pm, 2.1 × 100 mm, or an ACQUiTY BEH300 or BEH130 CT8 column, 1.77 pm, 2.1 × 100 mm, with a flow rate of 0.5 mL / min, λ-215 nm, 280 nm, and using 5–65% acetonitrile / water containing 0.05% TFA.

[0692] Method B, C18 HPLC conditions: HPLC analysis was performed on an Acquity BEH130, C18 column, 1.7 μm, 100 × 2.10 mm column at 25°C for 30 minutes with a flow rate of 0.5 mL / min, λ-215 nm, 280 nm, using 5–65% acetonitrile / water containing 0.05% TFA.

[0693] Method C, HPLC conditions: HPLC analysis was performed on an Acquity BEH130, C18 column, 1.7 μm, 100 × 2.10 mm column at 25°C for 20 minutes with a flow rate of 0.5 mL / min, λ-215 nm, 280 nm, using 5–65% acetonitrile / water containing 0.05% TFA.

[0694] Method D, HPLC conditions: HPLC analysis was performed on an Acquity BEH130, C18 column, 1.7 μm, 100 × 2.10 mm column at 25°C for 10 minutes at a flow rate of 0.5 mL / min, λ-215 nm, 280 nm, using 5–65% acetonitrile / water containing 0.05% TFA. 5.0 μL of sample was injected using PLNO (partial loop with needle overfill) injection mode.

[0695] Polypeptide analogs without lipophilic substituents and optional spacers may be referred to herein as “linear polypeptides.” Polypeptide analogs having at least one covalently bonded lipophilic substituent and optional spacers may be referred to herein as “conjugate polypeptides.” Table 13 provides characterization data for exemplary long-acting PYY analog polypeptides of this disclosure.

[0696] Polypeptide analogs that do not have lipophilic substituents and optional spacers may be referred to herein as “linear polypeptides.” Polypeptide analogs that have at least one covalently bonded lipophilic substituent and optional spacers may be referred to herein as “conjugate polypeptides.” [Table 15-1] [Table 15-2]

[0697] Example 3: Stability and solubility of long-acting PYY analog polypeptides The polypeptide analogs described herein were tested for solubility in physiological saline or water (DI water) at room temperature. The samples were visually inspected for clarity, turbidity, or cloudiness. The results of this analysis are shown in Table 14.

[0698] Several long-acting PYY analogs described herein were tested for stability in water (i.e., DI water) or physiological saline (1 mg / ml solution) as trifluoroacetates. These analog polypeptides were incubated at 37°C, and samples were collected at various time intervals. The purity and mass of the parent peptide, as well as the degree of degradation products, were analyzed by LC / MS and HPLC. The purity results from these analyses are shown in Table 14 and are considered indicators of stability. [Table 16]

[0699] Example 4: Human and rat NPY receptor function assay The NPY family of G protein-coupled receptors includes human and rat Y1, Y2, Y4, and Y5. Each receptor binds to a pharmacologically distinct set of endogenous peptide agonist hormones or neurotransmitters belonging to the NPY family, which include neuropeptide Y (NPY), peptide tyrosine-tyrosine (PYY), and pancreatic polypeptide (PP). NPY and PYY(1-36) bind to Y1, Y2, and Y5 receptors with relatively high affinity, while PYY(3-36) selectively binds to the Y2 receptor, and PP selectively binds to the Y4 receptor. Following agonist binding, the receptor-G protein complex activates downstream intracellular signaling cascades, resulting in inhibition of adenylyl cyclase and a decrease in intracellular cAMP levels.

[0700] Cell treatment and cAMP accumulation assay The functional efficacy of peptide agonists was characterized using the ActOne® membrane potential dye kit (Codex Biosolutions) with HEK-CNG cells (Codex Biosolutions) that stably express human or rat Y1, Y2, Y4, or Y5 receptors. In addition to each NPY receptor, each cell line encodes a unique exogenous cyclic nucleotide-gated (CNG) channel (Codex Biosolutions). The channel is activated by an increase in intracellular levels of cAMP, resulting in ion flux and cell membrane depolarization that can be detected with calcium-sensitive dyes or fluorescence membrane potential (MP) dyes. Cells were kept in growth medium containing 90% DMEM, 10% FBS, 250 mcg / mL G418, and 1 mcg / mL puromycin for 15 passages or less.

[0701] Prior to the experiment, cells were counted and dispensed into black 384-well poly-D-lysine plates at a rate of 20 mcL per well, with 14,000 cells per well (Y1, Y4, and Y5) or 28,000 cells per well (Y2). The covered plates were then incubated at room temperature for 30 minutes, followed by overnight incubation in 5% CO2 at 37°C. The following day, the medium was removed and the wells were washed once with 40 mcL of Dulbecco's phosphate-buffered saline. 20 microliters of DMEM, followed by 20 mcL of a dye loading solution containing 1X ACTOne® membrane potential dye dilution buffer, 1X ACTOne® membrane potential dye solution (Codex Biosolutions), and 50 mcM of phosphodiesterase inhibitor Ro20-1724, were added to each well. The plates were then covered and pre-incubated in the dark at room temperature for 2 hours. During this pre-incubation, experimental peptides and standards (NPY for Y1 and Y5, PYY(3-36) for Y2, and PP for Y4) were serially diluted to 12 different concentrations (ranging from 5x10⁻⁷M to 5x10⁻¹³M) in agonist dilution buffer containing 1X DPBS, 0.5% casein, 125mcM Ro20-1724, and 1.5mcM isoproterenol, stimulating b1 / b2 adrenergic receptor-mediated cAMP production. Following the pre-incubation, the first read (Ex530 / Em590) was performed using a Flexstation3 fluorescence plate reader (Molecular Devices, Sunnyvale, CA). Ten microliters of experimental peptide or peptide standard were added to the wells in triplicate and incubated in the dark at room temperature for 50 minutes. Following this incubation step, the plate was read again in the same manner as before (final read).

[0702] Data analysis and interpretation NPY receptor activation, which results in a decrease in intracellular cAMP levels, is detected as a reduction in isoproteranol-induced fluorescence signaling. The peptide standard and experimental values ​​were first converted in Excel using the formula: final reading / initial reading. The converted values ​​were then normalized to receptor-specific standard values ​​(minimum 1 × 10⁻¹³ M and maximum 1 × 10⁻⁷ M) using the formula: (test value - standard minimum mean) / (standard maximum mean - standard minimum mean) * 100. The normalized experimental values ​​represent the baseline-corrected percentage of the maximum response of the receptor system produced by the control peptide for each assay (NPY for Y1R and Y5R, PYY(3-36) for Y2R, and PP for Y4R). Analogues whose activity was ≤70% of the maximum value of the control peptide were identified as partial agonists. The normalized data were analyzed from three tests and used to estimate the EC50 of each test peptide for each receptor. The data was fitted using a 3-parameter logistic curve model: Y = bottom + (top - bottom) / (1 + 10^((LogEC50-X))) in GraphPad Prism software (v8.2.1). EC50 values ​​were converted to pEC50 values ​​using the formula: pEC50 = -Log(EC50). All reported values ​​are based on the curve fitting parameter r 2 The condition ≥ 0.8 was met.

[0703] The results of these analyses are reported in Table 15. [Table 17-1] [Table 17-2]

[0704] Example 5: In vitro metabolic stability pharmacokinetic study of PYY analog (T1 / 2) rat and human kidney brush border membrane In vitro incubation of renal brush border membrane (kBBM) preparations was used to characterize the ability of peptides to resist degradation by proteases and peptidases in systemic circulation. kBBM was chosen because it contains a diverse set of proteases and peptidases in high concentrations, many of which are present throughout the body. In general, peptides with low in vivo CL are stable in this assay, while peptides with high in vivo CL are unstable.

[0705] Brush border membranes from rat and human kidney tissue were prepared by centrifugation and stored at -70°C. Thawed rat or human kBBM stocks were diluted to appropriate concentrations in 25 mM HEPES buffer (pH 7.4) containing 1% casein and dispensed into 96-well plates. The kBBM solution was preheated at 37°C for 10 minutes. The reaction was initiated by adding the test peptide (final concentration 1 mcM) also dissolved in 25 mM HEPES buffer (pH 7.4) containing 1% casein. The final concentration of kBBM in each incubation was 50 mcg protein / mL. The reaction was maintained at 37°C in a shaking water bath. At 0, 0.25, 0.5, 1.0, 2.0, and 4.0 hours after initiation, 30 mcL of the reaction mixture was removed and placed in a 96-well plate containing 120 mcL of ice-cold methanol containing 2.5% formic acid. The quenched samples were centrifuged at 2178 × g for 10 minutes, and a portion of the supernatant was transferred to a clean 96-well plate and diluted 1:1 with water. The samples were analyzed by UPLC-MS / MS. The results of these analyses are shown in Tables 16, 17, and 18.

[0706] Human subcutaneous tissue homogenate Using in vitro incubation in subcutaneous (SC) tissue homogenates, we characterized the ability of peptides to resist pre-systemic degradation by proteases and peptidases after SC administration. In vivo nonclinical studies have shown that peptidase activity in the SC space may limit the bioavailability of peptides after SC administration. In this assay, peptides with high SC bioavailability are stable, while peptides with low SC bioavailability are unstable in this assay.

[0707] Human SC tissue was homogenized in 25 mM cold HEPES buffer (pH 7.4, 10 times the sample weight) and filtered through a double layer of cheesecloth. The filtrate was divided into equal portions, rapidly frozen in a methanol / dry ice bath, and stored at -80°C. The protein concentration of each pooled batch was determined using a BCA protein assay. The thawed human SC tissue homogenate stock was diluted to 1.0 mg protein / mL in 25 mM HEPES buffer (pH 7.4) and dispensed into 96-well plates. The diluted SC homogenate was preheated at 37°C for 10 minutes. The reaction was initiated by adding the test peptide (final concentration 10 mcM) also dissolved in 25 mM HEPES buffer (pH 7.4). The reaction was maintained at 37°C in a shaking water bath. At 0, 0.25, 0.5, 1.0, 2.0, and 4.0 hours after the start, 50 mcL of the reaction mixture was removed and placed in a 96-well plate containing 150 mcL of ice-cold methanol containing 2.5% formic acid. The quenched samples were centrifuged at 2178 × g for 10 minutes, and a portion of the supernatant was transferred to a clean 96-well plate and diluted 1:10 with water. The samples were analyzed by UPLC-MS / MS. The results of these analyses are shown in Tables 16, 17, and 18.

[0708] Unbound fraction in plasma (f u ) Because peptides tend to adsorb to the surfaces of plastic tubes, dialysis membranes, and molecular weight cutoff filters, conventional methods for measuring plasma protein binding, such as equilibrium dialysis, ultrafiltration, and ultracentrifugation, are unreliable. Therefore, the degree to which acylated peptides bind to serum albumin was assessed using surface plasmon resonance (SPR). The usefulness of this method for reasonably estimating the proportion of drugs bound to plasma proteins has been demonstrated in the literature. The results of this analysis for compounds A13 (SEQ ID NO: 24) and A24 (SEQ ID NO: 24) are shown in Tables 17 and 18. Based on this data, the estimated half-lives of compounds A13 and A24 in humans are approximately 4 days, respectively. [Table 18] [Table 19]

[0709] Example 6: Pharmacokinetic analysis of PYY analog polypeptides Intravenous infusion of long-acting PYY analog polypeptides to evaluate peptide clearance (CL). The peptide was dissolved in PBS with 0.05% Tween-20 (pH 7.4) and administered to non-fasted male Sprague-Dawley rats (n=3 per group) as a 1-hour subcutaneous infusion via a cannula placed in the femoral vein at a final dose of 0.033 mg / kg. The formulation was administered at a rate of 0.150 mL / hour / kg. Blood samples (approximately 250 μL) were collected via jugular vein cannula at 0.25, 0.5, 0.75, 1, 1.17, 1.33, 1.5, 2, 4, 8, 24, 48, 72, 96, and 120 hours after the start of infusion into microtinar tubes containing K2EDTA as an anticoagulant and 25 μL of a protease inhibitor cocktail for pharmacokinetic analysis. Plasma was prepared by centrifugation and stored at -80°C until analysis.

[0710] Subcutaneous bolus injection of long-acting PYY analog polypeptides to evaluate polypeptide bioavailability (F). The peptide was dissolved in sterile saline and administered to non-fasted male Sprague-Dawley rats (n=3 per group) via a single bolus injection into the subcutaneous space between the scapulae at a final dose of 0.1 mg / kg. Blood samples (approximately 250 μL) were collected via jugular vein cannula at 0.083, 0.25, 0.5, 1, 2, 4, 8, 24, 48, 72, 96, and 120 hours after administration into microtin tubes containing K2EDTA as an anticoagulant and 25 μL of a protease inhibitor cocktail for pharmacokinetic analysis. Plasma was prepared by centrifugation and stored at -80°C until analysis.

[0711] Plasma sample preparation for pharmacokinetic studies 70 μL aliquots of each plasma sample were placed in a 96-well plate. 210 μL of 0.1% TFA in a 2:1 ethanol:acetonitrile mixture containing a suitable internal standard was added to each well. The plate was vortexed at 1300 rpm for 10 minutes to mix, and then centrifuged at 500 × g for 10 minutes. The supernatant (210 μL) was placed in a clean 96-well plate and evaporated under a nitrogen stream at 45°C. The residue was reconstituted in 80 μL of 20% acetonitrile (aqueous solution) containing 0.1% formic acid.

[0712] LC / MS quantification of PYY analog polypeptides in plasma All calibration standards were prepared in control rat plasma containing K2EDTA and a protease inhibitor cocktail.

[0713] Samples and standards were analyzed by electrospray ionization (ESI) UPLC-MS / MS using a system consisting of an Agilent Infinity 1290 system (Palo Alto, CA) with a CTC HTS PAL automated injector (LeapCarrboro, NC), a column oven, a Valco switching valve (Houston, TX), and a Sciex TripleTOF® 5600 mass spectrometer (Framingham, MA). Samples were injected into a 2.1 × 50 mm reversed-phase C18 analytical column, typically Waters CORTECS UPLC C18+, 1.6 μm (Waters Corporation, Milford, MA) or similar. Chromatographic separation was performed using a gradient method with water (A) containing 0.1% formic acid and acetonitrile (B) containing 0.1% formic acid as mobile phases. The initial conditions consisted of 95% A and 5% B. Depending on the peptide, the organic component was increased to 95% B over 3–4 minutes. A typical flow rate was 550 μL / min. The column temperature was kept constant at 45°C. Peptides were quantified by monitoring one or more products generated from polyvalent parent ions. The results of these analyses are provided in Table 19. The comprehensive pharmacokinetic and ADME profiles of compounds A13 (SEQ ID NO: 13) and A24 (SEQ ID NO: 24) are shown in Tables 20 and 21, respectively. Changes in plasma concentrations of A13 and A24 after bolus and intravenous infusion are shown in Figures 4 and 5. [Table 20]

[0714] Example 7a: Weight loss efficacy of PYY analog in combination with a long-acting GLP-1 receptor agonist in LE DIO rats. To investigate the efficacy and durability of long-acting PYY analogs alone and in combination with long-acting GLP-1 analogs (LA GLP-1), a long-term weight loss efficacy study was conducted in a rodent model of obese Long Evans (LE) diet-induced obesity (DIO) rats. Male LE DIO rats (Envigo Laboratories, Inc., Indianapolis, IN) were used, starting from weaning, and were fed a high-fat diet (Teklad TD 95217, fat-derived, 40% kcal, Harlan Laboratories, Madison, WI). Rats were housed individually in cages, with free access to the high-fat diet (Harlan TD.95217) and water, maintained at 21°C and 50% relative humidity, in a 12-hour light / dark cycle from 5am to 5pm, and acclimatized for at least 10 days after shipment before use. At the start of the study, the rats were 16-18 weeks old. All procedures were carried out in accordance with the USDA Animal Welfare Act, which was approved by the Mispro Institutional Animal Care and Use Committee. Animals were randomly selected into treatment groups according to body weight and fat mass (n=8 rats / group). DIO LE rats were administered either a specified dose of long-acting PYY and / or a GLP-1 receptor agonist polypeptide or a vehicle control (saline) every other day (eod) via subcutaneous (SC) injection. The mean weight loss (%) ± SEM from baseline and vehicle control (ΔΔ) from long-term combined studies of compound A13 (SEQ ID NO: 13) and LA GLP-1 is shown in Figure 1. Comparative data for exenatide and semaglutide are shown in Figure 6.

[0715] Example 7b: Efficacy of weight gain of PYY analog administered alone to LE DIO rats In the experiment described in Example 7a, the response to a long-acting PYY agonist in combination with a long-acting GLP-1 agonist was analyzed, and LE DIO rats that received a long-acting PYY agonist (compound A24; SEQ ID NO: 24) alone were analyzed separately. Surprisingly, in contrast to the synergistic decrease in food intake and synergistic weight loss with the combination of compound A24 (SEQ ID NO: 24) and a long-acting GLP-1 receptor agonist, shown as the lower panel of Figure 8, the PYY agonist alone caused a dose-dependent increase in food intake and weight gain, as shown in the upper panel of Figure 8. When administered alone, another long-acting PYY agonist (compound A13; SEQ ID NO: 13) also showed a similar pattern of increased food intake and weight gain. The appetite-stimulating (food intake stimulation) and weight-gaining effects of both PYY agonists (A13 and A24) administered alone are in contrast to the previously described weight-change patterns with similarly selective (Y2) PYY agonists. This is the first example of a drug that is appetite-stimulating on its own but, when combined with a GLP-1 agonist, promotes a more potent appetite-suppressing effect than that observed with GLP-1 agonists alone.

[0716] The appetite-stimulating effects of SEQ ID NOs: 13 and 24 are not readily attributable to the long duration of action of these peptides themselves. For example, other selective PYY agonists whose duration of action is prolonged by PEGylation or binding to Fmoc showed only appetite-suppressing effects and no appetite-stimulating effects. Similarly, PYY-albumin conjugates did not show this effect and only showed appetite suppression, so the appetite-stimulating effects of these peptides are not readily attributable to albumin binding itself. Acylated PYY agonists that reversibly bind to albumin have not been previously reported to stimulate food intake and weight gain.

[0717] The appetite-stimulating effects of the PYY agonist class, represented by compounds A13 and A24 (see the upper left panel in Figure 8), are useful in certain conditions of pathogenic weight loss or decreased body energy. An example of such a condition is anorexia nervosa, a condition for which there are few pharmacological options and which is associated with approximately 10% mortality, mainly in young people. Another example is a form of cachexia associated with the treatment of malignant tumors, which complicates treatment. Yet another example is "frailty," or age-related nutritional deficiency. The components of this condition, represented by a decrease in muscle mass called sarcopenia, are involved in several age-related morbidities and a decline in quality of life, including loss of mobility, a tendency towards major bone fractures, and delayed healing of such fractures.

[0718] The therapeutically beneficial weight-gaining effects of the PYY agonist class, represented by compounds A13 and A24 (see upper right panel of Figure 8 and upper right quadrant of Figure 9), can be induced by administering these agents alone or in combination with other agents that produce beneficial effects through different mechanisms. Examples of distinct beneficial agents include appetite stimulants such as ghrelin and / or motilin agonists, or muscle growth-promoting agents such as anabolic steroids and growth hormone / inulin-like growth factor axis activators. Beneficial combinations for the above conditions do not include PYY agonists, represented by compounds A13 and A24, which are associated with GLP-1 receptor agonists, because such combinations are associated with weight loss (lower panel of Figure 8 and lower left quadrant of Figure 9).

[0719] Example 8: Antidiabetic efficacy of PYY analog in combination with GLP-1 agonist in ZDF rats A long-term study was conducted to determine the antidiabetic effect of continuous administration of a combination of PYY analog polypeptide and a GLP-1 receptor agonist on HbA1c (primary antidiabetic parameter) after 27 days of treatment in Zucker diabetic obese (ZDF) rats. Male ZDF rats were obtained at 6 weeks of age (Charles River, Raleigh, NC) and used in the study at 8 weeks of age. After receipt, rats were housed individually in cages, with free access to Purina 5008 diet (Lab Diet, St. Louis, MO) and water, maintained at 21°C and 50% relative humidity in a 12-hour light / dark cycle from 5am to 5pm, and acclimatized for 9 days before the start of the study. Blood samples were collected from the tail vein before blood collection (-3 days), and blood glucose and HbA1c levels were measured. ZDF rats were randomly selected into treatment groups (n=10 / group) with similar mean HbA1c and glucose levels. These ZDF rats were implanted subcutaneously (SC) with Alzet osmotic minipumps (2 pumps / animal) containing either a specified dose of PYY analog polypeptide and / or a GLP-1 receptor agonist (10 mcg / kg / day) or a vehicle (20% DMSO in water) (n=10 animals / treatment group). The PYY analog, with PK supporting alternating-day administration, was administered by SC injection instead of minipump administration. All other procedures were the same as described in the previous example. Blood samples were collected again on days 14 and 27 (end of the study) and blood glucose and HbA1c levels were measured. The final whole blood sample was collected by cardiac puncture under isoflurane anesthesia (day 27). HbA1c analysis was performed using a Carolina Chemistries CLC720i clinical chemistry analyzer (Mindray Inc., Mahwah, NY) using the protocol and method parameters described by the manufacturer. Figure 2 shows the HbA1c results, expressed as the mean percentage change from baseline and vehicle control (ΔΔ), from long-term combination studies of compound A13 (SEQ ID NO: 13) and a GLP-1 receptor agonist. Comparative data for tilzepatide and semaglutide are shown in Figure 7.

[0720] Other embodiments The present invention has been described in conjunction with its detailed description, but the foregoing description is intended to describe, not limit, the scope of the invention as defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

1. Amino acid sequence of SEQ ID NO: 90: X ０ PX ２ PX ４ X ５ PX ７ X ８ X ９ X １０ SPX １３ X １４ X １５ X １６ RX １８ X １９ X ２０ DX ２２ X ２３ HX ２５ X ２６ X ２７ WLTRX ３２ RX ３４ -(OH / NH ２ ) an isolated polypeptide comprising (SEQ ID NO: 90), or a pharmaceutically acceptable salt thereof (In the formula, X ０ is either nonexistent or K; X ２ is K; X ４ is either E or K; X ５ is A or K; X ７ is G or K; X ８ is E, K, or k; X ９ is either D or K; X １０ is A or K; X １３ is either E or K; X １４ is either E or K; X １５ is L or W; X １６ These are D, E, K, N, Q, S, T, α-methylserine, or homoserine; X １８ is either K or Y; X １９ is either K or Y; X ２０ is A, D, E, K, k, or Dap; X ２２ is A, D, K, or L; X ２３ is either K or R; X ２５ is either K or Y; X ２６ is E, K, or L; X ２７ is either K or N; X ３２ is either K or Q; X ３４ These are F, y, 3-pyridinylalanine, 4-pyridinylalanine, 4-carboxyphenylalanine, 4-fluorophenylalanine, 4-methylphenylalanine, N-methylphenylalanine, homophenylalanine, β-homotyrosine, homotyrosine, or N-methyltyrosine; Here, X ０ , X ２ , X ４ , X ５ , X ７ , X ８ , X １３ , X ２０ , X ２３ , X ２５ , X ２７ , or X ３２ When K is present, the lysine residue is optionally covalently bonded to a lipophilic substituent via a spacer. Here, X ８ or X ２０ When k, the D-lysine residue is optionally covalently bonded to a lipophilic substituent via a spacer. However, the polypeptide optionally includes at least one residue covalently bonded to a lipophilic substituent via a spacer; The polypeptide optionally further comprises lactam crosslinks formed via amide bonds between a lysine side chain and an aspartic acid side chain, or between a lysine side chain and a glutamic acid side chain, wherein the residues forming the lactam crosslinks are X ９ and X １３ position or X １６ and X ２０ position or X ２２ and X ２６ (To be in a certain position).

2. Amino acid sequence of SEQ ID NO: 91: PKPEX ５ PX ７ X ８ DASPX １３ EX １５ X １６ RYYX ２０ DX ２２ RHYLNWLTRQRX ３４ - (OH / NH ２ The isolated polypeptide according to claim 1, comprising (SEQ ID NO: 91), or a pharmaceutically acceptable salt thereof. (In the formula, X ５ is A or K; X ７ is G or K; X ８ is E, K, or k; X １３ is either E or K; X １５ is L or W; X １６ These are D, E, K, N, S, α-methylserine, or homoserine; X ２０ is A, D, E, K, or k; X ２２ is A or L; X ３４ These are F, 3-pyridinylalanine, 4-pyridinylalanine, 4-carboxyphenylalanine, 4-fluorophenylalanine, 4-methylphenylalanine, N-methylphenylalanine, homophenylalanine, β-homotyrosine, homotyrosine, or N-methyltyrosine; Here, X ５ , X ７ , X ８ , X １３ , or X ２０ When K is present, the lysine residue is optionally covalently bonded to a lipophilic substituent via a spacer. Here, X ８ or X ２０ is k, said D-lysine residue is optionally covalently bound to a lipophilic substituent, optionally via a spacer, However, the polypeptide optionally includes at least one residue covalently bonded to a lipophilic substituent via a spacer; The polypeptide optionally further comprises lactam crosslinks formed via amide bonds between a lysine side chain and an aspartic acid side chain, or between a lysine side chain and a glutamic acid side chain, wherein the residues forming the lactam crosslinks are X １６ and X ２０ (To be in a certain position).

3. Amino acid sequence of SEQ ID NO: 92: PKPEX ５ PX ７ X ８ DASPX １３ EX １５ X １６ RYYX ２０ DX ２２ RHYLNWLTRQRX ３４ -(OH / NH ２ ), which comprises SEQ ID NO: 92, the isolated polypeptide according to claim 1 or claim 2, or a pharmaceutically acceptable salt thereof (In the formula, X ５ is A or K; X ７ is G or K; X ８ is E, K, or k; X １３ is either E or K; X １５ is L or W; X １６ These are D, E, K, S, α-methylserine, or homoserine; X ２０ is A, D, E, K, or k; X ２２ is A or L; X ３４ These are F, 3-pyridinylalanine, 4-pyridinylalanine, 4-carboxyphenylalanine, 4-fluorophenylalanine, 4-methylphenylalanine, N-methylphenylalanine, homophenylalanine, β-homotyrosine, homotyrosine, or N-methyltyrosine; Here, X ５ , X ７ , X ８ , X １３ , or X ２０ When K is present, the lysine residue is optionally covalently bonded to a lipophilic substituent via a spacer. Here, X ８ or X ２０ When k, the D-lysine residue is optionally covalently bonded to a lipophilic substituent via a spacer. However, the polypeptide optionally includes at least one residue covalently bonded to a lipophilic substituent via a spacer; The polypeptide optionally further comprises lactam crosslinks formed via amide bonds between a lysine side chain and an aspartic acid side chain, or between a lysine side chain and a glutamic acid side chain, wherein the residues forming the lactam crosslinks are X １６ and X ２０ (To be in a certain position).

4. Amino acid sequence of SEQ ID NO: 93: PKPEX ５ PX ７ X ８ DASPX １３ EX １５ X １６ RYYX ２０ DX ２２ RHYLNWLTRQRX ３４ - (OH / NH ２ An isolated polypeptide according to claim 1 or claim 2, comprising (SEQ ID NO: 93), or a pharmaceutically acceptable salt thereof. (In the formula, X ５ is A or K; X ７ is G or K; X ８ is E, K, or k; X １３ is either E or K; X １５ is L or W; X １６ These are D, E, K, N, S, α-methylserine, or homoserine; X ２０ is D, E, K, or k; X ２２ is A or L; X ３４ These are F, 3-pyridinylalanine, 4-pyridinylalanine, 4-carboxyphenylalanine, 4-fluorophenylalanine, 4-methylphenylalanine, N-methylphenylalanine, homophenylalanine, β-homotyrosine, homotyrosine, or N-methyltyrosine; Here, X ５ , X ７ , X ８ , X １３ , or X ２０ When K is present, the lysine residue is optionally covalently bonded to a lipophilic substituent via a spacer. Here, X ８ or X ２０ When k, the D-lysine residue is optionally covalently bonded to a lipophilic substituent via a spacer. However, the polypeptide optionally includes at least one residue covalently bonded to a lipophilic substituent via a spacer; The polypeptide optionally further comprises lactam crosslinks formed via amide bonds between a lysine side chain and an aspartic acid side chain, or between a lysine side chain and a glutamic acid side chain, wherein the residues forming the lactam crosslinks are X １６ and X ２０ (To be in a certain position).

5. Amino acid sequence of SEQ ID NO: 94: PKPEX ５ PGX ８ DASPX １３ EWX １６ RYYX ２０ DX ２２ RHYLNWLTRQRX ３４ - (OH / NH ２ An isolated polypeptide according to claim 1 or claim 2, comprising (SEQ ID NO: 94), or a pharmaceutically acceptable salt thereof. (In the formula, X ５ is A or K; X ８ is E, K, or k; X １３ is either E or K; X １６ is D, E, K, or N; X ２０ is A, D, E, K, or k; X ２２ is A or L; X ３４ is F or N-methyltyrosine; Here, X ５ , X ８ , X １３ , or X ２０ When K is present, the lysine residue is optionally covalently bonded to a lipophilic substituent via a spacer. Here, X ８ or X ２０ When k, the D-lysine residue is optionally covalently bonded to a lipophilic substituent via a spacer. However, the polypeptide optionally includes at least one residue covalently bonded to a lipophilic substituent via a spacer; The polypeptide optionally further comprises lactam crosslinks formed via amide bonds between a lysine side chain and an aspartic acid side chain, or between a lysine side chain and a glutamic acid side chain, wherein the residues forming the lactam crosslinks are X １６ and X ２０ (To be in a certain position).

6. Amino acid sequence of SEQ ID NO: 95: PKPEX ５ PGK ８ DASPX １３ EWX １６ RYYX ２０ DLRHYLNWLTRQRF-(OH / NH ２ An isolated polypeptide according to any one of claims 1, 2, or 5, comprising (SEQ ID NO: 95), or a pharmaceutically acceptable salt thereof. (In the formula, X ５ is A or K; X １３ is either E or K; X １６ is D, E, K, or N; X ２０ is A, D, E, K, or k; Here, X ５ , X １３ or X ２０ When K is present, the lysine residue is optionally covalently bonded to a lipophilic substituent via a spacer. Here is the first K ８ It is optionally covalently bonded to a lipophilic substituent via a spacer, Here, X ２０ When k, the D-lysine residue is optionally covalently bonded to a lipophilic substituent via a spacer. However, the polypeptide optionally includes at least one residue covalently bonded to a lipophilic substituent via a spacer; The polypeptide optionally further comprises lactam crosslinks formed via amide bonds between a lysine side chain and an aspartic acid side chain, or between a lysine side chain and a glutamic acid side chain, wherein the residues forming the lactam crosslinks are X １６ and X ２０ (To be in a certain position).

7. X １５ If X is L, ２２ The isolated polypeptide according to any one of claims 1 to 4, wherein is A.

8. X １６ If X is N, ２０ The isolated polypeptide according to any one of claims 1, 2, or 5 to 7, wherein the polypeptide is not A.

9. X ８ E is X ５ K is X ２０ If X is K, ５ or X ２０ The isolated polypeptide according to any one of claims 1 to 5, 7, or 8, wherein any of the lysine residues is optionally covalently bonded to a lipophilic substituent via a spacer.

10. X ３４ The isolated polypeptide according to any one of claims 1 to 5 or 7 to 9, wherein F is present.

11. X ８ The isolated polypeptide according to any one of claims 1 to 5, 7, 8, or 10, wherein K is present.

12. The isolated polypeptide according to claim 6 or claim 11, wherein the lysine residue at position 8 of the polypeptide sequence is optionally covalently bonded to a lipophilic substituent via a spacer.

13. X １６ However, K is X ２０ However, it is D or E, and the polypeptide is X １６ and X ２０ The isolated polypeptide according to any one of claims 1 to 7 or 10 to 12, comprising a lactam crosslink formed via an amide bond between the side chains of the residue located at the position.

14. X １６ However, it is D or E, and X ２０ However, it is K, and the polypeptide is X １６ and X ２０ The isolated polypeptide according to any one of claims 1 to 7 or 10 to 12, comprising a lactam crosslink formed via an amide bond between the side chains of the residue located at the position.

15. An isolated polypeptide according to any one of claims 1 to 14, wherein an amide bond is formed between the amino group of the isolated polypeptide and the CO- group of the lipophilic substituent.

16. The aforementioned lipophilic substituent is of formula I: -CO-(CH ２ ) ｍ -Z Equation I The isolated polypeptide according to claim 15. (In the formula, Z is -CH ３ or -CO ２ It is H; m is between 4 and 24.

17. Z is -CO ２ The isolated polypeptide according to claim 16, wherein H.

18. The isolated polypeptide according to claim 16 or claim 17, wherein m is 14 to 20.

19. The isolated polypeptide according to any one of claims 16 to 18, wherein m is 16, 17, or 18.

20. An isolated polypeptide according to any one of claims 1 to 14, wherein the lipophilic substituent is covalently bonded to the isolated polypeptide via a spacer.

21. The isolated polypeptide according to claim 20, wherein the lipophilic substituent, i.e., -CO-(CH ２ ) ｍ -Z is linked to the amino group of the isolated polypeptide via the spacer, and the spacer forms a crosslink between the amino group of the isolated polypeptide and the CO- group of the lipophilic substituent, the isolated polypeptide.

22. The lipophilic substituent and the spacer are defined by formula II: -(Y) ｎ -CO-(CH ２ ) ｍ -Z Formula II The isolated polypeptide according to claim 20 or claim 21. (In the formula, Y is selected from the group consisting of γGlu, Asp, Lys, and Gly; Z is -CH ３ or -CO ２ It is H; m is between 4 and 24; n is between 1 and 10.

23. Z is -CO ２ The isolated polypeptide according to claim 22, wherein H is present.

24. The isolated polypeptide according to claim 22 or claim 23, wherein m is 14 to 20.

25. The isolated polypeptide according to any one of claims 22 to 24, wherein m is 16, 17, or 18.

26. The isolated polypeptide according to any one of claims 22 to 25, wherein Y is γGlu.

27. The isolated polypeptide according to any one of claims 22 to 26, wherein n is 1 to 5.

28. The isolated polypeptide according to any one of claims 22 to 27, wherein n is 2.

29. The lipophilic substituent and the spacer are: Formula IV: -(Y1) ｎ１ -(dpeg) ｒ -(Y2) ｎ２ -CO-(CH ２ ) ｍ -Z Formula IV The isolated polypeptide according to claim 20 or claim 21. (In the formula, Z is -CH ３ or -CO ２ It is H; m is between 4 and 24; Y1 is selected from the group consisting of γGlu, Asp, and Gly; Y2 is selected from the group consisting of γGlu, Asp, and Gly; dpeg is -[CO(CH ２ )O(CH ２ ) ２ O(CH ２ )NH]- and; r is between 1 and 8; n1 is between 0 and 10; n² is between 0 and 10.

30. Z is -CO ２ The isolated polypeptide according to claim 29, wherein H.

31. The isolated polypeptide according to claim 29 or claim 30, wherein m is 14 to 20.

32. The isolated polypeptide according to any one of claims 29 to 31, wherein m is 16, 17, or 18.

33. The isolated polypeptide according to any one of claims 29 to 32, wherein Y1 is γGlu.

34. An isolated polypeptide according to any one of claims 29 to 33, wherein n1 is 0 to 3.

35. The isolated polypeptide according to any one of claims 29 to 34, wherein Y2 is γGlu.

36. An isolated polypeptide according to any one of claims 29 to 35, wherein n2 is 0 to 3.

37. An isolated polypeptide according to any one of claims 29 to 36, wherein r is 1 to 3.

38. The carboxy-terminal amino acid, namely X ３４ However, -X ３４ - (NH ２ The isolated polypeptide according to any one of claims 1 to 37.

39. An isolated polypeptide containing any of the amino acid sequences of SEQ ID NOs: 1 to 78, or a pharmaceutically acceptable salt thereof.

40. The isolated polypeptide according to claim 39, comprising an amino acid sequence selected from the group consisting of those represented by SEQ ID NOs: 13, 24, 42, and 43.

41. An isolated polypeptide according to claim 39, comprising the amino acid sequence of SEQ ID NO: 13, or a pharmaceutically acceptable salt thereof.

42. An isolated polypeptide according to claim 39, comprising the amino acid sequence of SEQ ID NO: 24, or a pharmaceutically acceptable salt thereof.

43. An isolated polypeptide according to claim 39, comprising the amino acid sequence of SEQ ID NO: 42, or a pharmaceutically acceptable salt thereof.

44. An isolated polypeptide according to claim 39, comprising the amino acid sequence of SEQ ID NO: 43, or a pharmaceutically acceptable salt thereof.

45. A pharmaceutical composition comprising an isolated polypeptide according to any one of claims 1 to 44, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable adjuvant, carrier, or vehicle.

46. A pharmaceutical combination comprising an isolated polypeptide according to any one of claims 1 to 44, or a pharmaceutically acceptable salt thereof, and a GLP-1 receptor agonist.

47. An osmotic delivery device comprising an isolated polypeptide according to any one of claims 1 to 44, a pharmaceutical composition according to claim 45, or a pharmaceutical combination according to claim 46.

48. An impermeable reservoir including its inner and outer surfaces and first and second open ends; A semipermeable membrane that is in a sealed relationship with the first open end of the reservoir; An osmotic engine located within the reservoir and adjacent to the semipermeable membrane; A piston adjacent to the osmotic engine, wherein the piston forms a movable seal with the inner surface of the reservoir, and the piston divides the reservoir into a first chamber and a second chamber, the first chamber containing the osmotic engine, and the piston; A suspension formulation wherein the second chamber contains the suspension formulation, the suspension formulation is fluid, and contains the isolated polypeptide; The osmotic delivery device according to claim 47, comprising a diffusion moderator inserted into the second open end of the reservoir, the diffusion moderator adjacent to the suspension formulation.

49. A method for treating obesity in a human subject, causing weight loss in the human subject, or suppressing the appetite of the human subject, comprising administering to the subject a pharmaceutical composition comprising an isolated polypeptide according to any one of claims 1 to 44, the pharmaceutical composition according to claim 45, the pharmaceutical combination according to claim 46, or the osmotic delivery device according to claim 47 or 48.

50. A method for treating diabetes in a human subject, comprising administering to the subject a pharmaceutical composition comprising an isolated polypeptide according to any one of claims 1 to 44, a pharmaceutical composition according to claim 45, a pharmaceutical combination according to claim 46, or an osmotic delivery device according to claim 47 or 48.

51. The method according to claim 50, wherein the diabetes is type 1 diabetes.

52. The method according to claim 50, wherein the diabetes is type 2 diabetes.

53. A method for treating non-alcoholic fatty liver disease (NAFLD) and / or non-alcoholic steatohepatitis (NASH) in a human subject, comprising administering to the subject a pharmaceutical composition comprising an isolated polypeptide according to any one of claims 1 to 44, the pharmaceutical composition according to claim 45, the pharmaceutical combination according to claim 46, or the osmotic delivery device according to claim 47 or 48.

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