RU2839688C2 - Human amylin analog polypeptides and methods of use thereof - Google Patents

Abstract

FIELD: biotechnology; medicine.

SUBSTANCE: present invention relates to biotechnology and can be used in medicine. Invention discloses novel human amylin analogues having a number of advantages in comparison with endogenous amylin, for example, increased half-life (t_1/2), as well as improved solubility and thermal stability.

EFFECT: disclosed amylin analogue polypeptides can be used in treating metabolic diseases or disorders, such as type 1 and 2 diabetes, and in providing body weight loss.

20 cl, 3 dwg, 11 tbl, 17 ex

Description

This application claims the benefit of and the right to benefit from U.S. Provisional Patent Application No. 62/744,236, filed October 11, 2018, which is incorporated herein by reference in its entirety.

List of sequences

This application contains a sequence listing filed electronically in ASCII format and incorporated herein by reference in its entirety. The ASCII copy created on October 10, 2019 is named 616782_102487-055PC_SL-10-10-2019.txt and is 123,647 bytes in size.

AREA OF TECHNOLOGY

The present invention relates to isolated polypeptides that are analogs of human amylin. The disclosed amylin analog polypeptides have advantageous physicochemical properties relative to endogenous amylin, such as longer half-lives (t _1/2 ) and improved solubility and thermal stability. The present invention also relates to methods of using the amylin analog polypeptides disclosed herein in various therapeutic indications, as well as methods for their preparation. As explained in more detail below, the disclosed amylin analog polypeptides are particularly useful in methods of treating metabolic diseases or disorders, such as type 1 and type 2 diabetes, as well as in providing weight loss.

LEVEL OF TECHNOLOGY

Human amylin, or islet amyloid polypeptide (IAPP), is a 37-residue polypeptide hormone. Amylin is cosecreted with insulin from pancreatic β-cells 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, 7,404-dalton propeptide that undergoes post-translational modifications, including protease cleavage, to yield the 37-residue amylin. Loss of β-cell function, which occurs early in type 1 diabetes and may persist late in type 2 diabetes, results in deficient secretion of insulin and amylin.

Amylin functions as part of the endocrine pancreas, the cells in the pancreas that synthesize and secrete hormones. Amylin contributes to glycemic control; it is secreted from the pancreatic islets into the circulation and cleared by peptidases in the kidneys. The metabolic function of amylin is well characterized as an inhibitor of the release of nutrients such as glucose into the plasma. It thus functions as a synergistic partner to insulin, a peptide that regulates blood glucose levels and coordinates glucose uptake and distribution in the body. Insulin's role in the body is, among other things, to prevent blood glucose levels from rising too high, especially after meals.

Amylin is thought to play a role in glycemic regulation by slowing gastric emptying and promoting satiety (i.e., feeling full), thereby preventing postprandial (i.e., after eating) blood glucose peaks. The overall effect is to slow the rate at which glucose appears in the blood after a meal. Amylin also reduces the secretion of glucagon by the pancreas. Glucagon's role in the body is, among other things, to prevent blood glucose levels from getting too low. This is important because certain type 1 diabetics, for example, are prone to secreting excess glucagon, which raises blood glucose immediately after eating.

For numerous reasons, human amylin, which has a serum half-life of approximately 13 minutes, is not amenable to use as a therapeutic agent. Rather, pramlintide (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 patients with type 1 or type 2 diabetes who use mealtime insulin but are unable to achieve desired glycemic control despite optimal insulin therapy. Pramlintide differs from human amylin in 3 of 37 amino acids. These modifications provide pramlintide with a longer half-life of approximately 48 minutes and reduce its aggregation potential characteristic of human amylin.

For the treatment of type 1 diabetes, pramlintide is administered up to four times daily, by subcutaneous injection before meals, as an adjunct to post-meal insulin therapy. Pramlintide cannot be mixed with insulin; separate syringes are used. Reported side effects of pramlintide include nausea and vomiting. Adverse reactions may include serious hypoglycemia, particularly for type 1 diabetes. Consequently, mealtime insulin dosage is reduced for patients who initiate pramlintide.

For the treatment of type 2 diabetes, pramlintide is administered before each meal by subcutaneous injection at the recommended starting dose, which is gradually increased to the target technical dose. Another investigational human amylin analogue, davalintide (AC2307; also developed by Amylin Pharmaceuticals, Inc.), has a half-life of approximately 26 minutes.

Accordingly, there is a need for improved amylin analog polypeptides that mimic amylin activity yet have greater therapeutic potential than both endogenous human amylin and existing amylin analogs such as primalintide and davalintide.

ESSENCE OF THE INVENTION

The polypeptides of the present invention and pharmaceutically acceptable compositions thereof have been found to be effective as amylin analogs. Such polypeptides have the general formula SEQ ID NO: 199:

X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TX ₃₇ -(OH/NH ₂ ) (SEQ ID NO: 199), or a pharmaceutically acceptable salt thereof, wherein:

X ₁ is S, K, k, H, or I; X ₃ is N or S; X ₅ is S or A; X ₆ is T or S; X ₈ is A or K; X ₁₀ is Q or S; X ₁₂ is L or K; X ₁₃ is A, S, E or K; X ₁₄ is N, n, d, Y or Q; X ₁₅ is E, F, f, Y, I, k, K, or α-aminoisobutyric acid (Aib); X ₁₆ is k, K, L, Aib, N-methyl leucine (N-MeL), or 1; X ₁₇ is H, V, Q, R, k, K, or Aib; X ₁₈ is K, H, or R; X ₁₉ is S or Aib; X ₂₀ is S or Aib; X ₂₂ is N or E; X ₂₉ represents P, R, or K; X ₃₁ represents k, K, N, or H; X ₃₅ represents e, E, N, K, G, A, Y, or P, and X ₃₇ represents Y or P;

each K is independently L-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

each k is independently D-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

wherein two cysteine residues X ₁ CX ₃ TX ₅ X ₆ C are optionally additionally linked by a disulfide bridge;

provided that if X ₃₁ is N then X ₃₅ is E, or if X ₃₅ is N then X ₃₁ is K.

Brief description of graphic materials

Fig. 1 is a table illustrating comparative sequence alignments for certain reference polypeptides: human amylin, SEQ ID NO: 300; rat amylin, SEQ ID NO: 301; pramlintide, SEQ ID NO: 302; davalintide, SEQ ID NO: 303; hCT (human calcitonin), SEQ ID NO: 304; sCT (salmon calcitonin), SEQ ID NO: 305, and beta-calcitonin gene-related peptide (β-CGRP), SEQ ID NO: 306.

Figs. 2A, 2B, 2C, and 2D show dose response curves for an acylated amylin A73 analog at hAMY3R (Fig. 2A), an acylated amylin A73 analog at hCTR (Fig. 2B), pramlintide at hAMY3R (Fig. 2C), and human calcitonin (hCalcitonin) at hCTR (Fig. 2D). The term "acylated" as used herein with respect to the disclosed polypeptides means that the disclosed polypeptide is substituted with one or more lipophilic substituents that are each optionally attached through a spacer, wherein "lipophilic substituent" and "spacer" are defined herein.

Figures 3A and 3B show data from pharmacokinetic studies assessing polypeptide clearance from the kidneys (CL) following intravenous infusion of linear, i.e., non-acylated, polypeptides (Figure 3A) and conjugated, i.e., acylated, polypeptides (Figure 3B).

DETAILED DESCRIPTION OF THE ESSENCE OF THE INVENTION

1. General description of certain embodiments of the invention

The present invention relates to isolated polypeptides that are amylin analogs, as well as pharmaceutical compositions containing these polypeptides. The present invention also relates to methods for producing and using such amylin analog polypeptides. These amylin analog polypeptides are particularly useful in methods for treating metabolic diseases or disorders, such as type 1 and type 2 diabetes, obesity, and in methods for providing weight loss.

2. Definitions

It is understood that the terminology used herein is for the purpose of describing particular embodiments of the invention only and is not intended to limit the scope of the disclosure. Use in this specification and the appended claims of the singular includes plural reference unless the context clearly dictates otherwise. Thus, for example, reference to a "solvent" includes a combination of two or more such solvents, reference to a "peptide" includes one or more peptides or a mixture of peptides, reference to a "drug" includes one or more drugs, reference to an "osmotic delivery device" includes one or more osmotic delivery devices, and the like. Unless specifically indicated or obvious from the context, when used herein, the term "or" is understood to include and encompass both "or" and "and".

Unless otherwise stated or obvious from the context, when used herein, the term "about" is understood to mean "within the normal tolerance range" in the art, such as within 2 standard deviations of the mean. "About" or "approximately" 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 clear from the context, all numerical values presented herein are modified by the term "about" or "approximately."

Unless otherwise stated or obvious from context, when used herein, the term "substantially" shall be understood to mean "within a narrow range of variation" or otherwise within normal tolerance in the art. Substantially may be understood to mean within 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, 0.01%, or 0.001% of the stated value.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although other methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, the materials and methods described herein are preferred.

In the description and claims of this invention, the following terminology will be used in accordance with the definitions set forth below.

The terms "drug", "therapeutic agent" and "beneficial substance" are used interchangeably to refer to any therapeutically active substance that is delivered to a subject to obtain a desired beneficial effect. In one embodiment of the invention, the drug is a polypeptide. In another embodiment of the invention, the drug is a small molecule, such as hormones such as androgens or estrogens. The devices and methods of the invention are well suited for delivering proteins, small molecules and combinations thereof.

The terms "peptide", "polypeptide" and "protein" are used interchangeably herein and generally refer to a molecule comprising a chain 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 mimetics).

In some embodiments, the naturally occurring L-amino acids are represented by either the conventional three-letter or capital single-letter amino acid designations from Table 1. In other embodiments, the naturally occurring L-amino acids and D-amino acids are the conventional three-letter or capital single-letter amino acid designations from Table 1. In other embodiments, the D-amino acids are represented by the lowercase single-letter designation corresponding to the single-letter designations from Table 1, i.e., g, a, 1, m, f, w, k, q, e, s, p, v, i, c, y, h, r, n, d, and t.

Peptides may be naturally occurring, synthetic, or recombinantly expressed. Peptides may also contain additional groups that modify the amino acid chains, such as functional groups added by post-translational modification. Examples of post-translational modifications include, but are not limited to, acetylation, alkylation (including methylation), biotinylation, glutamylation, glycylation, glycosylation, isoprenylation, lipoylation, phosphopantetheinylation, phosphorylation, selenium, and C-terminal amidation. The term "peptide" also includes peptides containing modifications at the amino terminus and/or carboxy terminus. Modifications at the terminal amino group include, but are not limited to, deamination, N-lower alkyl, N-di-lower alkyl, and N-acyl modifications. Modifications of the carboxyl group of the terminal include, but are not limited to, modifications involving an amide, a lower alkyl amide, a dialkyl amide, and a lower alkyl ester (e.g., where the lower alkyl is C1-C4 alkyl). The term "peptide" also includes modifications, such as, but not limited to, those described above, of the amino acids located between the amino and carboxy termini. In one embodiment, the peptide can be modified by the addition of a small molecule drug.

The terminal amino acid at one end of a peptide chain typically has a free amino group (e.g., the amino terminus). The terminal amino acid at the other end of the chain typically has a free carboxyl group (e.g., the carboxy terminus). Typically, the amino acids that make up a peptide are numbered in order starting from the amino terminus with increasing numbers toward the carboxy terminus of the peptide.

The phrase "amino acid residue" as used herein refers to an amino acid that is incorporated into a peptide via an amide bond or an amide bond mimetic.

The term "insulinotropic" as used herein generally refers to the ability of a compound, such as a peptide, to stimulate or affect the production and/or activity of insulin (e.g., insulinotropic hormone). Such compounds generally stimulate or otherwise affect the secretion or biosynthesis of insulin in a subject. Thus, an "insulinotropic peptide" is a molecule containing an amino acid that is capable of stimulating or otherwise affecting the secretion or biosynthesis of insulin.

The term "insulinotropic peptide" as used herein includes, but is not limited to, glucagon-like peptide 1 (GLP-1) and its derivatives and analogs, GLP-1 receptor agonists such as exenatide, exenatide of the amino acid sequence SEQ ID NO: 307, and their derivatives and analogs.

The term "acylated" as used herein with respect to the disclosed polypeptides means that the disclosed polypeptide is substituted with one or more lipophilic substituents, each optionally attached through a spacer, wherein "lipophilic substituent" and "spacer" are defined herein. Certain lipophilic substituents, each optionally via a spacer, can bind albumin and exhibit affinity for albumin with the resulting acylated polypeptide. The extent is variable and depends on numerous factors to which the lipophilic substituents, each optionally via a spacer, bind albumin and exhibit affinity for albumin with the resulting acylated polypeptide. Numerous factors include the particulars of the lipophilic substituent, the optional spacer, the polypeptide, and the site of covalent attachment to the polypeptide.

The terms "linear" or "linear polypeptide" as used herein refer to a "non-acylated" polypeptide, in other words, the disclosed amylin polypeptide analogs without lipophilic substituents, each optionally attached via a spacer, wherein "lipophilic substituent" and "spacer" are defined herein.

The terms "conjugated" or "conjugated polypeptide" as used herein refers to an "acylated" polypeptide, in other words, the disclosed amylin polypeptide analog, having one or more lipophilic substituents, each optionally attached through a spacer, wherein "lipophilic substituent" and "spacer" are defined herein.

As used herein, the term "pharmaceutically acceptable salt" refers to those salts that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic reaction, etc., and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of this invention include salts prepared from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable acid addition salts are salts of the amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or with organic acids such as acetic acid, trifluoroacetic acid (TFA), oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid, or by other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citran, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc.

Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(C1-4 alkyl)4 salts. Typical alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Further pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium and amine cations formed with counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate.

The phrase "incretin mimetics" as used herein includes, but is not limited to, a GLP-1 peptide, a GLP-1 receptor agonist, GLP-1 peptide derivatives, GLP-1 peptide analogs; exenatide, exenatide having the amino acid sequence of SEQ ID NO: 307, exenatide peptide, exenatide peptide derivatives, and exenatide peptide analogs. Examples of preferred incretin mimetics include exenatide, exenatide with the amino acid sequence exendin-4 (the natural form of exenatide), exenatide-LAR, lixisenatide, GLP-1(7-36), liraglutide, semaglutide, dulaglutide, aluiglutide, and taspoglutide. Incretin mimetics are also referred to herein as "insulinotropic peptides." Incretin mimetics that target GLP-1 receptors are also known in the literature as "GLP-1 receptor agonists" or "GLP-1 agonists," with both terms used interchangeably herein.

The term "exenatide" as used herein includes, but is not limited to, exenatide, exenatide having the amino acid sequence (HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS- _NH2 ), SEQ ID NO: 307, naturally occurring exendin-4, exenatide peptide, peptide analogs of exenatide, and peptide derivatives of exenatide.

The term "GLP-1" refers to a polypeptide that is produced by L cells located primarily in the ileum and colon and to a lesser extent by L cells in the duodenum and jejunum. GLP-1 is a regulatory peptide that binds to the extracellular region of the GLP-1 receptor (GLP-1R), a G protein-coupled receptor on the β-cell and, through adenylyl cyclase activity and AMP production, stimulates the insulin response to nutrients absorbed from the intestine [Baggio 2007, "Biology of incretins: GLP-1 and GIP," Gastroenterology, vol. 132(6):2131–57; Hoist 2008, "The incretin system and its role in type 2 diabetes mellitus," Mol Cell Endocrinology, vol. 297(1–2): 127–36]. The effects of GLP-1R agonism are multiple. GLP-1 maintains glucose homeostasis by enhancing endogenous glucose-dependent insulin secretion, rendering β-cells glucose-competent and GLP-1-sensitive, suppressing glucagon release, restoring the first and second phases of insulin secretion, slowing gastric emptying, decreasing food intake, and increasing satiety [Hoist 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; Hoist 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]. The risk of hypoglycemia is minimal given the mode of action of GLP-1.

As described in more detail below, in some embodiments, the amylin analog polypeptides disclosed herein are provided in methods for treating type 1 diabetes, as an adjunct to insulin treatment. The term "insulin" as used herein refers to human insulin or any insulin analogs. Exemplary non-limiting insulin analogs include those listed in Table 2:

As used herein, the term "mealtime insulin" refers to a rapid-acting insulin formulation that reaches peak blood concentrations approximately 45 to 90 minutes and peak activity approximately 1 to 3 hours after administration and is administered at or near the time of a meal.

The term "carrier" as used herein refers to a medium used to carry a compound, such as a drug or a particle containing a drug. The carriers of the present invention typically contain components such as polymers and solvents. The suspension carriers of the present invention typically contain solvents and polymers that are used to prepare suspension compositions that further contain drug particle formulations.

The phrase "phase separation" as used herein refers to the formation of multiple phases (e.g., a liquid phase and a gel phase) in a suspension carrier, such as upon contact of the suspension carrier with an aqueous medium. In some embodiments of the present invention, the suspension carrier is manufactured such that it exhibits phase separation upon contact with an aqueous medium containing less than about 10% water.

The phrase "single-phase" as used in this description refers to a solid, semi-solid, or liquid homogeneous system that is physically and chemically uniform throughout its volume.

The term "dispersed" as used herein refers to dissolving, dispersing, suspending or otherwise distributing a compound, such as a drug, in a suspension vehicle.

The phrase "chemically stable" as used herein refers to the formation of an acceptable percentage of degradation products in the composition over a specified period of time by chemical pathways such as deamidation (usually by hydrolysis), aggregation, or oxidation.

The phrase "physically stable" as used herein refers to the formation of an acceptable percentage of aggregates (e.g., dimers and other higher molecular weight products) in the composition. In addition, a physically stable composition does not change its physical state, such as from a liquid to a solid or from an amorphous to a crystalline form.

The term "viscosity" as used herein generally refers to a value determined based on the ratio of shear stress to shear rate (see, e.g., Considine, D. M. & Considine, G. D., Encyclopedia of Chemistry, 4th Edition, Van Nostrand, Reinhold, N. Y., 1984), essentially as follows:

where F/A = shear stress (force per unit area),

μ = proportionality coefficient (viscosity), and

V/L = velocity versus layer thickness (shear rate).

Based on this relationship, the ratio of shear stress to shear rate determines the viscosity. Shear stress and shear rate measurements are typically determined using a parallel plate rheometer, the rheometry being performed under selected conditions (e.g., at a temperature of about 37°C). Other methods for determining viscosity include measuring kinematic viscosity using viscometers, such as a Cannon-Fenske viscometer, an Ubbelohde viscometer in a Cannon-Fenske opaque solution, or an Ostwald viscometer. Typically, the suspension carriers of the present invention have a viscosity sufficient to prevent sedimentation of particles suspended therein during storage and use in a delivery method, such as an implantable drug delivery device.

The term "non-aqueous" as used herein refers to a total moisture content, for example in a suspension composition, typically less than or equal to about 10 wt.%, for example less than or equal to about 7 wt.%, less than or equal to about 5 wt.%, and/or less than about 4 wt.% In addition, the particle composition of the present invention contains less than about 10 wt.%, for example less than about 5 wt.%, residual moisture.

The term "subject" as used herein refers to any member of the subphylum Chordata, including, without limitation, humans and other primates, including non-human primates such as rhesus macaques and other monkeys, chimpanzees and other apes; farm animals such as cattle, sheep, pigs, goats and horses; domestic animals such as dogs and cats; laboratory animals, including rodents such as mice, rats and guinea pigs; birds, including domestic, wild and game birds such as chickens, turkeys and other members of the order Galliformes, ducks, geese and the like. The term does not denote a specific age or sex. Thus, both adults and neonates are intended to be covered.

As used herein, the terms "treatment," "treat," and "treatment process" refer to reversing, alleviating, delaying the onset, or inhibiting the progress of a disease or disorder or multiple symptoms thereof described herein. In some embodiments, treatment may be administered after one or more symptoms have developed. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a predisposed patient prior to the onset of symptoms (e.g., based on a history of symptoms and/or based on genetic or other predisposing factors). Treatment may also be continued after symptoms have resolved, for example, to prevent or slow their recurrence.

The term "osmotic delivery device" as used herein generally refers to a device used to deliver a drug (e.g., a disclosed amylin analog polypeptide) to a subject, wherein the device includes, for example, a reservoir (made, for example, of a titanium alloy) having a lumen that contains a suspension formulation containing a drug (e.g., a disclosed amylin analog polypeptide) and an osmotic agent formulation. A piston assembly located in the lumen isolates the suspension formulation from the osmotic agent formulation. A semipermeable membrane located at a first distal end of the reservoir is adjacent to the osmotic agent formulation; a diffusion retarder (which defines a delivery orifice through which the suspension formulation exits the device) is located at a second distal end of the reservoir and is adjacent to the suspension formulation. Typically, the osmotic delivery device is implanted in a subject, for example, subdermally or subcutaneously (e.g., in the inner part, outer part, or back of the upper arm and in the abdominal region). An example of an osmotic delivery device is the DUROS® delivery device (ALZA Corporation, Mountain View, CA). Examples of synonyms for the terms "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.

The term "continuous delivery" as used herein generally refers to a substantially continuous release of a drug from an osmotic delivery device into tissue near the implantation site, such as subdermal and subcutaneous tissue. For example, an osmotic delivery device releases a drug at a substantially set rate based on the principle of osmosis. Extracellular fluid enters the osmotic delivery device through a semipermeable membrane directly into an osmotic motor, which expands to drive a piston at a low and stable rate of movement. The movement of the piston expels the drug formulation through the orifice of the diffusion retarder. Thus, the drug is released from the osmotic delivery device at a low, controlled and stable rate.

The term "substantially stable delivery" as used herein generally refers to delivery of a drug at or near a target concentration over a specified period of time, wherein the amount of drug delivered from the osmotic delivery device is substantially zero-order delivery. Substantially zero-order delivery of an active substance (e.g., the disclosed amylin analog polypeptide) means that the rate of drug delivery is constant and independent of the drug available in the delivery system; for example, for zero-order delivery, if the rate of drug delivery versus time is plotted on a graph and the plotted line fits the data, the line has a slope near zero as determined by standard methods (e.g., linear regression).

The phrase "half-life of a drug" as used in this document refers to how long it takes for a drug to be eliminated from the blood plasma to half its concentration. The half-life of a drug is typically measured by monitoring the decline in drug levels when the drug is administered by injection or intravenously. The drug is typically measured using, for example, a radioimmunoassay (RIA), a chromatographic assay, an electrochemiluminescence (ECL) assay, an enzyme-linked immunosorbent assay (ELISA), or a sandwich enzyme-linked immunosorbent assay (SELISA).

The term "µg" should be understood as "micrograms". Similarly, the term "µL" should be understood as "microliter" and the term "µM" should be understood as "µmol/L".

The term "serum" is intended to refer to any blood product in which a substance can be detected. Thus, the term "serum" includes at least whole blood, serum, and plasma. For example, "the amount of [a substance] in a subject's serum" would include "the amount of [a substance] in a subject's plasma."

Baseline is defined as the last assessment on or before the day of primary placement of the osmotic delivery device (containing drug or placebo).

3. Endogenous amylin, related peptides and amylin receptors

Human amylin is a 37-residue polypeptide hormone cosecreted with insulin from pancreatic β-cells. Loss of β-cell function, which occurs early in type 1 diabetes and may occur late in type 2 diabetes, results in inadequate secretion of insulin and amylin. Amylin is thought to play a role in glycemic regulation by slowing gastric emptying and promoting satiety, thereby preventing postprandial peaks in blood glucose levels. The overall effect is a slowing of the rate of appearance of glucose in the blood after a meal.

The amino acid sequence of amylin is most closely related to that of calcitonin gene-related peptide (CGRP). CGRP also has a similarly positioned disulfide bond and an amidated C-terminus. This is also characteristic of calcitonin, adrenomedullin, and adrenomedulin 2. Together, these peptides form a small family united by these characteristics. Consequently, there is a degree of overlap in the related receptor binding for each peptide and in the pharmacologic activity. The table in Fig. 1 illustrates comparative sequence alignments for amylin and certain related reference polypeptides.

Peptides commonly designated as calcitonin (CT) family member peptides include calcitonin gene-related peptide (CGRP), calcitonin (CT), amylin (AMY), adrenomedullin 1, and adrenomedullin 2/intermedin (ADM1, ADM2, respectively). Two G protein-coupled receptor proteins (calcitonin receptor; CTR and calcitonin-like receptor receptor; CALCRL) and three receptor activity modifying proteins (RAMP1, RAMP2, RAMP3) constitute the pharmacologically distinct receptors for the entire peptide family (CTR, AMY1, AMY2, AMY3, CGRPR, AM1, AM2). There appear to be at least five different receptors to which amylin binds with significant affinity (AMY1, AMY2, AMY3, CTR, CGRPR). CTR dimerizes with RAMP 1, 2, or 3 to restore the AMY1, AMY2, or AMY3 receptors with amylin-selective pharmacology over calcitonin. In the absence of RAMP, CTR pharmacology becomes calcitonin-selective over amylin. CALCRL dimerized with RAMP1 creates CGRPR with high affinity for CGRP and reduced affinity for all other members of the peptide family, including amylin. CALCRL and RAMP2 or RAMP3, restore AM1 and AM2 pharmacology, respectively, with very low or no affinity for amylin.

Analogs of the amylin polypeptide that have binding affinity for the amylin receptor complexes have been developed. Pramlintide, for example, was developed by Amylin Pharmaceuticals and approved by the U.S. Food and Drug Administration (FDA) as a synthetic analog of human amylin for the treatment of type 1 and type 2 diabetics who use mealtime insulin but fail to achieve desired glycemic control despite optimal insulin therapy. Pramlintide is an amylin mimetic agent that has at least comparable activity to human amylin. It is also a 37-amino acid polypeptide, and differs in amino acid sequence from human amylin by substitution of amino acids with proline at positions 25 (alanine), 28 (serine), and 29 (serine). As a result of these substitutions, pramlintide is soluble, non-sticky, and non-aggregating, thereby overcoming a number of the physicochemical deficiencies of native human amylin. The half-life of pramlintide is approximately 48 minutes in humans, longer than that of native human amylin (approximately 13 minutes). Pramlintide requires frequent and inconvenient administration.

For the treatment of type 1 diabetes, pramlintide is administered up to four times daily by subcutaneous injection into the thigh or abdomen before meals, as an adjunct to postprandial insulin therapy. Pramlintide cannot be mixed with insulin; separate syringes are used. Pramlintide is administered before each meal or snack containing at least 250 calories or 30 g of carbohydrate. The typical starting dose for type 1 diabetes is 15 mcg of pramlintide subcutaneously before each meal, titrated to a target dose of 60 mcg before each meal. Reported adverse effects of pramlintide include nausea and vomiting. Adverse events, particularly for type 1 diabetes, may include severe hypoglycemia. Consequently, mealtime insulin dosage is reduced for patients with diabetes who initiate pramlintide.

For the treatment of type 2 diabetes, pramlintide is administered by subcutaneous injection at a recommended starting dose of 60 mcg, with a target maintenance dose of 120 mcg before each meal.

Davalintide (AC2307) is another human amylin analogue. Davalintide is an experimental compound with a half-life of approximately 26 minutes. Like pramlintide, davalintide also requires frequent administration by injection.

Certain disclosed amylin analog polypeptides, including those listed in Table 3 below, exhibit one or more of: superior solubility, stability, biological activity and specificity, and a longer half-life than endogenous human amylin and known synthetic amylin analog polypeptides. Certain disclosed amylin analog polypeptides were designed to accommodate less frequent administrations than required for pramlintide. Certain disclosed amylin analog polypeptides were designed to be administered via weekly or monthly injections. Certain disclosed amylin analog polypeptides were designed to be administered via implantation of a delivery device comprising an amylin analog polypeptide, wherein the delivery device contains a dose of the amylin analog polypeptide for 3 months, 6 months, 9 months, one year, 18 months, or two years.

4. Description of Illustrative Embodiments

In certain embodiments, the present invention relates to isolated polypeptides that are amylin analogs.

In some embodiments, an isolated polypeptide of the invention comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus sequence SEQ ID NO: 199:

X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TX ₃₇ -(OH/NH ₂ ) (SEQ ID NO: 199), or a pharmaceutically acceptable salt thereof, wherein:

X ₁ represents S, K, k, H or I;

X ₃ represents N or S;

X ₅ represents S or A;

X ₆ represents T or S;

X ₈ represents A or K;

X ₁₀ represents Q or S;

X ₁₂ represents L or K;

X ₁₃ represents A, S, E or K;

X ₁₄ represents N, n, d, Y or Q;

X ₁₅ is E, F, f, Y, I, k, K, or α-aminoisobutyric acid (Aib);

X ₁₆ represents k, K, L, Aib, N-methylleucine (N-MeL), or 1;

X ₁₇ represents H, V, Q, R, k, K or Aib;

X ₁₈ represents K, H, or R;

X ₁₉ represents S or Aib;

X ₂₀ represents S or Aib;

X ₂₂ represents N or E;

X ₂₉ represents P, R or K;

X ₃₁ represents k, K, N, or H;

X ₃₅ represents e, E, N, K, G, A, Y, or P, and

X ₃₇ represents Y or P;

each K is independently L-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

each k is independently D-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

wherein two cysteine residues X ₁ CX ₃ TX ₅ X ₆ C are optionally additionally linked by a disulfide bridge;

provided that if X ₃₁ is N then X ₃₅ is E, or if X ₃₅ is N then X ₃₁ is K.

In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the specific amino acids represented by the consensus sequence SEQ ID NO: 199 include the following:

In some embodiments, the carboxy-terminal X ₃₇ is Y-(NH ₂ ). In some embodiments, the carboxy-terminal X ₃₇ is Y-(OH). In some embodiments, the carboxy-terminal X ₃₇ is P-(NH ₂ ). In some embodiments, the carboxy-terminal X ₃₇ is P-(OH).

In some embodiments, X ₁ is S. In some embodiments, X ₁ is K. In some embodiments, X ₁ is k. In some embodiments, X ₁ is H. In some embodiments, X ₁ is I.

In the context of this document, k refers to D-lysine.

In some embodiments, X ₃ is N. In some embodiments, X ₃ is S.

In some embodiments, X ₅ is S. In some embodiments, X ₅ is A.

In some embodiments, X ₆ is T. In some embodiments, X ₆ is S.

In some embodiments, X ₈ is A. In some embodiments, X ₈ is K.

In some embodiments, X ₁₀ is Q. In some embodiments, X ₁₀ is S.

In some embodiments, X ₁₂ is L. In some embodiments, X ₁₂ is K.

In some embodiments, X ₁₃ is A. In some embodiments, X ₁₃ is S. In some embodiments, X ₁₃ is E. In some embodiments, X ₁₃ is K.

In some embodiments, X ₁₄ is N. In some embodiments, X ₁₄ is n. In some embodiments, X ₁₄ is d. In some embodiments, X ₁₄ is Y. In some embodiments, X ₁₄ is Q.

In the context of this document, n denotes D-asparagine.

In the context of this document, d refers to D-aspartic acid.

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F. In some embodiments, X ₁₅ is f. In some embodiments, X ₁₅ is Y. In some embodiments, X ₁₅ is I. In some embodiments, X ₁₅ is K. In some embodiments, X ₁₅ is k. In some embodiments, X ₁₅ is Aib.

In the context of this document, f denotes D-phenylalanine.

In the context of this document, Aib means alternatively 2-aminoisobutyric acid, α-aminoisobutyric acid, α-methylalanine or 2-methylalanine.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is I. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k. In some embodiments, X ₁₆ is Aib. In some embodiments, X ₁₆ is N-MeL.

In the context of this document, 1 refers to D-leucine.

In the context of this document, N-MeL stands for N-methylleucine.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q. In some embodiments, X ₁₇ is R. In some embodiments, X ₁₇ is K. In some embodiments, X ₁₇ is k. In some embodiments, X ₁₇ is Aib.

In some embodiments, X ₁₈ is K. In some embodiments, X ₁₈ is H. In some embodiments, X ₁₈ is R.

In some embodiments, X ₁₉ is S. In some embodiments, X ₁₉ is Aib.

In some embodiments, X ₂₀ is S. In some embodiments, X ₂₀ is Aib.

In some embodiments, X ₂₂ is N. In some embodiments, X ₂₂ is E.

In some embodiments, X ₂₉ is P. In some embodiments, X ₂₉ is R. In some embodiments, X ₂₉ is K.

In some embodiments, X ₃₁ is k. In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N. In some embodiments, X ₃₁ is H.

In some embodiments, X ₃₅ is e. In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N. In some embodiments, X ₃₅ is K. In some embodiments, X ₃₅ is G. In some embodiments, X ₃₅ is A. In some embodiments, X ₃₅ is Y. In some embodiments, X ₃₅ is P.

In the context of this document, e stands for D-glutamic acid.

In some embodiments, the specific amino acids represented by the consensus sequence SEQ ID NO: 199 include the following:

In some embodiments, X ₁ is S and X ₅ is S. In some embodiments, X ₁ is S and X ₁₀ is Q. In some embodiments, X ₁ is S and X ₁₅ is E. In some embodiments, X ₁ is S and X ₁₆ is L. In some embodiments, X ₁ is S and X ₁₆ is k. In some embodiments, X ₁ is S and X ₁₇ is H. In some embodiments, X ₁ is S and X ₁₈ is K. In some embodiments, X ₁ is S and X ₃₁ is K. In some embodiments, X ₁ is S and X ₃₅ is E. In some embodiments, X ₁ is S and X ₃₇ is Y.

In some embodiments, X ₁ is K and X ₅ is S. In some embodiments, X ₁ is K and X ₁₀ is Q. In some embodiments, X ₁ is K and X ₁₅ is E. In some embodiments, X ₁ is K and X ₁₆ is L. In some embodiments, X ₁ is K and X ₁₆ is k. In some embodiments, X ₁ is K and X ₁₇ is H. In some embodiments, X ₁ is K and X ₁₈ is K. In some embodiments, X ₁ is K and X ₃₁ is K. In some embodiments, X ₁ is K and X ₃₅ is E. In some embodiments, X ₁ is K and X ₃₇ is Y.

In some embodiments, X ₁ is k and X ₅ is S. In some embodiments, X ₁ is k and X ₁₀ is Q. In some embodiments, X ₁ is k and X ₁₅ is E. In some embodiments, X ₁ is k and X ₁₆ is L. In some embodiments, X ₁ is k and X ₁₆ is k. In some embodiments, X ₁ is k and X ₁₇ is H. In some embodiments, X ₁ is k and X ₁₈ is K. In some embodiments, X ₁ is k and X ₃₁ is K. In some embodiments, X ₁ is k and X ₃₅ is E. In some embodiments, X ₁ is k and X ₃₇ is Y.

In some embodiments, X ₅ is S and X ₁₅ is E. In some embodiments, X ₅ is S and X ₁₀ is Q. In some embodiments, X ₅ is S and X ₁₆ is L. In some embodiments, X ₅ is S and X ₁₆ is k. In some embodiments, X ₅ is S and X ₁₇ is H. In some embodiments, X ₅ is S and X ₁₈ is K. In some embodiments, X ₅ is S and X ₃₁ is K. In some embodiments, X ₅ is S and X ₃₅ is E. In some embodiments, X ₅ is S and X ₃₇ is Y.

In some embodiments, X ₁₀ is Q and X ₁₅ is E. In some embodiments, X ₁₀ is Q and X ₁₆ is L. In some embodiments, X ₁₀ is Q and X ₁₆ is k. In some embodiments, X ₁₀ is Q and X ₁₇ is H. In some embodiments, X ₁₀ is Q and X ₁₈ is K. In some embodiments, X ₁₀ is Q and X ₃₁ is K. In some embodiments, X ₁₀ is Q and X ₃₅ is E. In some embodiments, X ₁₀ is Q and X ₃₇ is Y.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₈ is K. In some embodiments, X ₁₅ is E and X ₃₁ is K. In some embodiments, X ₁₅ is E and X ₃₅ is E. In some embodiments, X ₁₅ is E and X ₃₇ is Y.

In some embodiments, X ₁₀ is L and X ₁₇ is H. In some embodiments, X ₁₀ is L and X ₁₈ is K. In some embodiments, X ₁₀ is L and X ₃₁ is K. In some embodiments, X ₁₀ is L and X ₃₅ is E. In some embodiments, X ₁₀ is L and X ₃₇ is Y.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₈ is K. In some embodiments, X ₁₆ is k and X ₃₁ is K. In some embodiments, X ₁₆ is k and X ₃₅ is E. In some embodiments, X ₁₆ is k and X ₃₇ is Y.

In some embodiments, X ₁₇ is H and X ₁₈ is K. In some embodiments, X ₁₇ is H and X ₃₁ is K. In some embodiments, X ₁₇ is H and X ₃₅ is E. In some embodiments, X ₁₇ is H and X ₃₇ is Y.

In some embodiments, X ₁₈ is K and X ₃₁ is K. In some embodiments, X ₁₈ is K and X ₃₅ is E. In some embodiments, X ₁₈ is K and X ₃₇ is Y.

In some embodiments, X ₃₁ is K and X ₃₅ is E. In some embodiments, X ₃₁ is K and X ₃₇ is Y.

In some embodiments, X ₃₅ is E and X ₃₇ is Y.

In some embodiments, the specific amino acids represented by the consensus sequence SEQ ID NO: 199 include the following:

In some embodiments, X ₁ is S, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is S, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is S, and X ₃₅ is E. In some embodiments, X ₁ is S, X ₅ is S, and X ₃₇ is Y.

In some embodiments, X ₁ is K, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₅ is E. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₇ is Y.

In some embodiments, X ₁ is k, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is k, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is k, X ₅ is S, and X ₃₅ is E. In some embodiments, X ₁ is k, X ₅ is S, and X ₃₇ is Y.

In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₆ is L. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₆ is k. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₇ is H. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₈ is K. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₃₁ is K. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₃₅ is E. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₃₇ is Y.

In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₁₆ is L. In some embodiments, X ₁₀ is Q, and X ₁₆ is k, and X ₁₇ is H. In some embodiments, X ₁₀ is Q, X ₁₈ is K, and X ₃₁ is K. In some embodiments, X ₁₀ is Q, X ₃₁ is K, and X ₃₅ is E. In some embodiments, X ₁₀ is Q, X ₃₁ is K, and X ₃₇ is Y.

In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₅ is E. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₇ is Y.

In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₅ is E. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₇ is Y.

In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₅ is E. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₇ is Y.

In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₅ is E. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₇ is Y.

In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₁ is K. In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₅ is E. In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₇ is Y.

In some embodiments, X ₁₈ is K, X ₃₁ is K, and X ₃₅ is E. In some embodiments, X ₁₈ is K, X ₃₁ is K, and X ₃₇ is Y.

In some embodiments, X ₃₁ is K, X ₃₅ is E, and X ₃₇ is Y.

In some embodiments, carboxy-terminal amino acid 37 is Y-(NH ₂ ). In some embodiments, carboxy-terminal amino acid 37 is Y-(OH). In some embodiments, carboxy-terminal amino acid 37 is P-(NH ₂ ). In some embodiments, carboxy-terminal amino acid 37 is P-(OH).

In some embodiments, an isolated polypeptide of the invention comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus sequence SEQ ID NO: 200:

X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 200), or a pharmaceutically acceptable salt thereof, wherein:

X ₁ represents S, K, k, H or I;

X ₃ represents N or S;

X ₅ represents S or A;

X ₆ represents T or S;

X ₈ represents A or K;

X ₁₀ represents Q or S;

X ₁₂ represents L or K;

X ₁₃ represents A, S, E or K;

X ₁₄ represents N, n, d, Y or Q;

X ₁₅ is E, F, f, Y, I, k, K, or α-aminoisobutyric acid (Aib);

X ₁₆ represents k, K, L, Aib, N-methylleucine (N-MeL), or 1;

X ₁₇ represents H, V, Q, R, k, K or Aib;

X ₁₈ represents K, H, or R;

X ₁₉ represents S or Aib;

X ₂₀ represents S or Aib;

X ₂₂ represents N or E;

X ₂₉ represents P, R or K;

X ₃₁ represents k, K, or N; and

X ₃₅ represents e, E, or N;

each K is independently L-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

each k is independently D-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

wherein two cysteine residues X ₁ CX ₃ TX ₅ X ₆ C are optionally additionally linked by a disulfide bridge;

provided that if X ₃₁ is N then X ₃₅ is E, or if X ₃₅ is N then X ₃₁ is K.

In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, an isolated polypeptide of the invention comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus sequence of SEQ ID NO: 201: X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TKVGSETY-(OH/NH ₂ ) (SEQ ID NO: 201), wherein:

X ₁ represents S, K, k, H, or I;

X ₃ represents N or S;

X ₅ represents S or A;

X ₆ represents T or S;

X ₈ represents A or K;

X ₁₀ represents Q or S;

X ₁₂ represents L or K;

X ₁₃ represents A, S, E or K;

X ₁₄ represents N, n, d, Y or Q;

X15 represents E, F, f, Y, I, k, K, or Aib;

X ₁₆ represents k, K, L, Aib, N-MeL, or 1;

X ₁₇ represents H, V, Q, R, k, K or Aib;

X ₁₈ represents K, H, or R;

X ₁₉ represents S or Aib;

X ₂₀ represents S or Aib;

X ₂₂ represents N or E, and

X ₂₉ represents P, R or K;

each K is independently L-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

each k is independently D-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

wherein two cysteine residues X ₁ CX ₃ TX ₅ X ₆ C are optionally additionally linked by a disulfide bridge.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 200 or SEQ ID NO: 201 include the following:

In some embodiments, carboxy-terminal amino acid 37 is Y-(NH ₂ ). In some embodiments, carboxy-terminal amino acid 37 is Y-(OH).

In some embodiments, X ₁ is S. In some embodiments, X ₁ is K. In some embodiments, X ₁ is k. In some embodiments, X ₁ is H. In some embodiments, X ₁ is I.

In the context of this document, k refers to D-lysine.

In some embodiments, X ₃ is N. In some embodiments, X ₃ is S.

In some embodiments, X ₅ is S. In some embodiments, X ₅ is A.

In some embodiments, X ₆ is T. In some embodiments, X ₆ is S.

In some embodiments, X ₈ is A. In some embodiments, X ₈ is K.

In some embodiments, X ₁₀ is Q. In some embodiments, X ₁₀ is S.

In some embodiments, X ₁₂ is L. In some embodiments, X ₁₂ is K.

In some embodiments, X ₁₃ is A. In some embodiments, X ₁₃ is S. In some embodiments, X ₁₃ is E. In some embodiments, X ₁₃ is K.

In some embodiments, X ₁₄ is N. In some embodiments, X ₁₄ is n. In some embodiments, X ₁₄ is d. In some embodiments, X ₁₄ is Y. In some embodiments, X ₁₄ is Q.

In the context of this document, n denotes D-asparagine.

In the context of this document, d refers to D-aspartic acid.

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F. In some embodiments, X ₁₅ is f. In some embodiments, X ₁₅ is Y. In some embodiments, X ₁₅ is I. In some embodiments, X ₁₅ is K. In some embodiments, X ₁₅ is k. In some embodiments, X ₁₅ is Aib.

In the context of this document, f denotes D-phenylalanine.

In the context of this document, Aib means alternatively 2-aminoisobutyric acid, α-aminoisobutyric acid, α-methylalanine or 2-methylalanine.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is I. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k. In some embodiments, X ₁₆ is Aib. In some embodiments, X ₁₆ is N-MeL.

In the context of this document, 1 refers to D-leucine.

In the context of this document, N-MeL stands for N-methylleucine.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q. In some embodiments, X ₁₇ is R. In some embodiments, X ₁₇ is K. In some embodiments, X ₁₇ is k. In some embodiments, X ₁₇ is Aib.

In some embodiments, X ₁₈ is K. In some embodiments, X ₁₈ is H. In some embodiments, X ₁₈ is R.

In some embodiments, X ₁₉ is S. In some embodiments, X ₁₉ is Aib.

In some embodiments, X ₂₀ is S. In some embodiments, X ₂₀ is Aib.

In some embodiments, X ₂₂ is N. In some embodiments, X ₂₂ is E.

In some embodiments, X ₂₉ is P. In some embodiments, X ₂₉ is R. In some embodiments, X ₂₉ is K.

In some embodiments, X ₃₁ is k. In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is e. In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In the context of this document, e stands for D-glutamic acid.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 200 or SEQ ID NO: 201 include the following:

In some embodiments, X ₁ is S and X ₅ is S. In some embodiments, X ₁ is S and X ₁₀ is Q. In some embodiments, X ₁ is S and X ₁₅ is E. In some embodiments, X ₁ is S and X ₁₀ is L. In some embodiments, X ₁ is S and X ₁₀ is k. In some embodiments, X ₁ is S and X ₁₇ is H. In some embodiments, X ₁ is S and X ₁₀ is K. In some embodiments, X ₁ is S and X ₃₁ is K. In some embodiments, X ₁ is S and X ₃₅ is E.

In some embodiments, X ₁ is K and X ₅ is S. In some embodiments, X ₁ is K and X ₁₀ is Q. In some embodiments, X ₁ is K and X ₁₅ is E. In some embodiments, X ₁ is K and X ₁₆ is L. In some embodiments, X ₁ is K and X ₁₆ is k. In some embodiments, X ₁ is k and X ₁₇ is H. In some embodiments, X ₁ is k and X ₁₈ is K. In some embodiments, X ₁ is K and X ₃₁ is K. In some embodiments, X ₁ is K and X ₃₅ is E.

In some embodiments, X ₁ is k and X ₅ is S. In some embodiments, X ₁ is k and X ₁₀ is Q. In some embodiments, X ₁ is k and X ₁₅ is E. In some embodiments, X ₁ is k and X ₁₆ is L. In some embodiments, X ₁ is k and X ₁₆ is k. In some embodiments, X ₁ is k and X ₁₇ is H. In some embodiments, X ₁ is k and X ₁₈ is K. In some embodiments, X ₁ is k and X ₃₁ is K. In some embodiments, X ₁ is k and X ₃₅ is E.

In some embodiments, X ₅ is S and X ₁₅ is E. In some embodiments, X ₅ is S and X ₁₀ is Q. In some embodiments, X ₅ is S and X ₁₆ is L. In some embodiments, X ₅ is S and X ₁₆ is k. In some embodiments, X ₅ is S and X ₁₇ is H. In some embodiments, X ₅ is S and X ₁₈ is K. In some embodiments, X ₅ is S and X ₃₁ is K. In some embodiments, X ₅ is S and X ₃₅ is E.

In some embodiments, X ₁₀ is Q and X ₁₅ is E. In some embodiments, X ₁₀ is Q and X ₁₆ is L. In some embodiments, X ₁₀ is Q and X ₁₆ is k. In some embodiments, X ₁₀ is Q and X ₁₇ is H. In some embodiments, X ₁₀ is Q and X ₁₈ is K. In some embodiments, X ₁₀ is Q and X ₃₁ is K. In some embodiments, X ₁₀ is Q and X ₃₅ is E.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₈ is K. In some embodiments, X ₁₅ is E and X ₃₁ is K. In some embodiments, X ₁₅ is E and X ₃₅ is E.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₈ is K. In some embodiments, X ₁₆ is L and X ₃₁ is K. In some embodiments, X ₁₆ is L and X ₃₅ is E.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₈ is K. In some embodiments, X ₁₆ is k and X ₃₁ is K. In some embodiments, X ₁₆ is k and X ₃₅ is E.

In some embodiments, X ₁₇ is H and X ₁₈ is K. In some embodiments, X ₁₇ is H and X ₃₁ is K. In some embodiments, X ₁₇ is H and X ₃₅ is E.

In some embodiments, X ₁₈ is K and X ₃₁ is K. In some embodiments, X ₁₈ is K and X ₃₅ is E.

In some embodiments, X ₃₁ is K and X ₃₅ is E.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 200 or SEQ ID NO: 201 include the following:

In some embodiments, X ₁ is S, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is S, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is S, and X ₃₅ is E.

In some embodiments, X ₁ is K, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₅ is E.

In some embodiments, X ₁ is k, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is k, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is k, X ₅ is S, and X ₃₅ is E.

In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₆ is L. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₆ is k. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₇ is H. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₈ is K. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₃₁ is K. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₃₅ is E.

In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₁₆ is L. In some embodiments, X ₁₀ is Q, and X ₁₆ is k, and X ₁₇ is H. In some embodiments, X ₁₀ is Q, X ₁₈ is K, and X ₃₁ is K. In some embodiments, X ₁₀ is Q, X ₃₁ is K, and X ₃₅ is E.

In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₅ is E.

In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₅ is E.

In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₅ is E.

In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₅ is E.

In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₁ is K. In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₅ is E.

In some embodiments, X ₁₈ is K, X ₃₁ is K, and X ₃₅ is E.

In some embodiments, an isolated polypeptide of the invention comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus sequence SEQ ID NO: 202 X ₁ CNTX ₅ TCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ X ₁₈ SSNNFGPILPPTX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 202), wherein:

X ₁ represents S, k or K;

X ₅ represents S or A;

X ₁₀ represents Q or S;

X ₁₅ represents E or F;

X ₁₆ represents k, K or L;

X ₁₇ represents H, V and Q;

X ₁₈ represents K, H, or R;

X ₃₁ represents K or N; and

X ₃₅ represents E or N;

each K is independently L-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

each k is independently D-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

wherein the two cysteine residues X ₁ CNTX ₅ TC (SEQ ID NO: 308) are optionally further linked by a disulfide bridge; and

provided that if X ₃₁ is N then X ₃₅ is E, or if X ₃₅ is N then X ₃₁ is K.

In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, an isolated polypeptide of the invention comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus sequence of SEQ ID NO: 203: X ₁ CNTX ₅ TCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ X ₁₈ SSNNFGPILPPTKVGSETY-(OH/NH ₂ ) (SEQ ID NO: 203), wherein:

X ₁ represents S, k or K;

X ₅ represents S or A;

X ₁₀ represents Q or S;

X ₁₅ represents E or F;

X ₁₆ represents k, K or L;

X ₁₇ represents H, V and Q; and

X ₁₈ represents K, H, or R;

each K is independently L-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

each k is independently D-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

wherein the two cysteine residues X ₁ CNTX ₅ TC (SEQ ID NO: 308) are optionally further linked by a disulfide bridge.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 202 or SEQ ID NO: 203 include the following:

In some embodiments, carboxy-terminal amino acid 37 is Y-(NH ₂ ). In some embodiments, carboxy-terminal amino acid 37 is Y-(OH).

In some embodiments, X ₁ is S. In some embodiments, X ₁ is K. In some embodiments, X ₁ is k.

In some embodiments, X ₅ is S. In some embodiments, X ₅ is A.

In some embodiments, X ₁₀ is Q. In some embodiments, X ₁₀ is S.

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q.

In some embodiments, X ₁₈ is K. In some embodiments, X ₁₈ is H. In some embodiments, X ₁₈ is R.

In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 202 or SEQ ID NO: 203 include the following:

In some embodiments, X ₁ is S and X ₅ is S. In some embodiments, X ₁ is S and X ₁₀ is E. In some embodiments, X ₁ is S and X ₁₆ is L. In some embodiments, X ₁ is S and X ₁₆ is k. In some embodiments, X ₁ is S and X ₁₇ is H. In some embodiments, X ₁ is S and X ₁₈ is K. In some embodiments, X ₁ is S and X ₃₁ is K. In some embodiments, X ₁ is S and X ₃₅ is E.

In some embodiments, X ₁ is K and X ₅ is S. In some embodiments, X ₁ is K and X ₁₅ is E. In some embodiments, X ₁ is K and X ₁₆ is L. In some embodiments, X ₁ is K and X ₁₆ is k. In some embodiments, X ₁ is K and X ₁₇ is H. In some embodiments, X ₁ is k and X ₁₈ is K. In some embodiments, X ₁ is K and X ₃₁ is K. In some embodiments, X ₁ is K and X ₃₅ is E.

In some embodiments, X ₅ is S and X ₁₅ is E. In some embodiments, X ₅ is S and X ₁₆ is L. In some embodiments, X ₅ is S and X ₁₆ is k. In some embodiments, X ₅ is S and X ₁₇ is H. In some embodiments, X ₅ is S and X ₁₈ is K. In some embodiments, X ₅ is S and X ₃₁ is K. In some embodiments, X ₅ is S and X ₃₅ is E.

In some embodiments, X ₁₀ is Q and X ₁₅ is E. In some embodiments, X ₁₀ is Q and X ₁₆ is L. In some embodiments, X ₁₀ is Q and X ₁₇ is k. In some embodiments, X ₁₀ is Q and X ₁₇ is H. In some embodiments, X ₁₀ is Q and X ₁₈ is K. In some embodiments, X ₁₀ is Q and X ₃₁ is K. In some embodiments, X ₁₀ is Q and X ₃₅ is E.

In some embodiments, X ₁₅ is E and X ₁₀ is L. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₈ is K. In some embodiments, X ₁₅ is E and X ₃₁ is K. In some embodiments, X ₁₅ is E and X ₃₅ is E.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₈ is K. In some embodiments, X ₁₆ is L and X ₃₁ is K. In some embodiments, X ₁₆ is L and X ₃₅ is E.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₈ is K. In some embodiments, X ₁₆ is k and X ₃₁ is K. In some embodiments, X ₁₆ is k and X ₃₅ is E.

In some embodiments, X ₁₇ is H and X ₁₈ is K. In some embodiments, X ₁₇ is H and X ₃₁ is K. In some embodiments, X ₁₇ is H and X ₃₅ is E.

In some embodiments, X ₁₈ is K and X ₃₁ is K. In some embodiments, X ₁₈ is K and X ₃₅ is E.

In some embodiments, X ₃₁ is K and X ₃₅ is E.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 202 or SEQ ID NO: 203 include the following:

In some embodiments, X ₁ is S, X ₅ is S, and X ₁₀ is Q. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₈ is k. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is S, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is S, and X ₃₅ is E.

In some embodiments, X ₁ is K, X ₅ is S, and X ₁₀ is Q. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is K, X ₅ is S, and X _1e is k. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₅ is E.

In some embodiments, X ₅ is S, X ₁₀ is Q, and X ₁₅ is E. In some embodiments, X ₅ is S, X ₁₀ is Q, and X ₁₆ is L. In some embodiments, X ₅ is S, X ₁₀ is Q, and X ₁₀ is k. In some embodiments, X ₅ is S, X ₁₀ is Q, and X ₁₇ is H. In some embodiments, X ₅ is S, X ₁₀ is Q, and X ₁₀ is K. In some embodiments, X ₅ is S, X ₁₀ is Q, and X ₃₁ is K. In some embodiments, X ₅ is S, X ₁₀ is Q, and X ₃₅ is E.

In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₁₆ is L. In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₁₀ is k. In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₁₇ is H. In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₁₈ is K. In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₃₁ is K. In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₃₅ is E.

In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₀ is L, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₀ is L, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₀ is L, and X ₃₅ is E.

In some embodiments, X ₁₅ is E, X ₁₀ is k, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₀ is k, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₀ is k, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₀ is k, and X ₃₅ is E.

In some embodiments, X ₁₀ is L, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₀ is L, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₅ is E.

In some embodiments, X ₁ e is k, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₅ is E.

In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₁ is K. In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₅ is E.

In some embodiments, X ₁₈ is K, X ₃₁ is K, and X ₃₅ is E.

In some embodiments, an isolated polypeptide of the invention comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus sequence of SEQ ID NO: 204: X ₁ CNTSTCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 204), wherein:

X ₁ represents S, K or k;

X ₁₀ represents Q or S;

X ₁₅ represents E or F;

X ₁₆ represents L, K or k;

X ₁₇ represents H, V, or Q; and

X ₃₅ represents E or N;

each K is independently L-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

each k is independently D-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

wherein the two cysteine residues X ₁ CNTSTC (SEQ ID NO: 309) are optionally further linked by a disulfide bridge.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, an isolated polypeptide of the invention comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus sequence of SEQ ID NO: 205: X ₁ CNTSTCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSETY-(OH/NH ₂ ) (SEQ ID NO: 205), wherein:

X ₁ represents S, K or k;

X ₁₀ represents Q or S;

X ₁₅ represents E or F;

X ₁₀ represents L, K, or k; and

X ₁₇ represents H, V or Q;

each K is independently L-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

each k independently represents D-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer; and

wherein the two cysteine residues X ₁ CNTSTC (SEQ ID NO: 309) are optionally further linked by a disulfide bridge.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 204 or SEQ ID NO: 205 include the following:

In some embodiments, carboxy-terminal amino acid 37 is Y-(NH ₂ ). In some embodiments, carboxy-terminal amino acid 37 is Y-(OH).

In some embodiments, X ₁ is S. In some embodiments, X ₁ is K. In some embodiments, X ₁ is k.

In some embodiments, X ₁₀ is Q. In some embodiments, X ₁₀ is S.

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 204 or SEQ ID NO: 205 include the following:

In some embodiments, X ₁ is S and X ₁₀ is Q. In some embodiments, X ₁ is S and X ₁₀ is S. In some embodiments, X ₁ is S and X ₁₅ is E. In some embodiments, X ₁ is S and X ₁₅ is F. In some embodiments, X ₁ is S and X ₁₆ is L. In some embodiments, X ₁ is S and X ₁₆ is K. In some embodiments, X ₁ is S and X ₁₆ is k. In some embodiments, X ₁ is S and X ₁₇ is H. In some embodiments, X ₁ is S and X ₁₇ is V. In some embodiments, X ₁ is S and X ₁₇ is Q. In some embodiments, X ₁ is S and X ₃₅ is E. In some embodiments, X ₁ is S and X ₃₅ is N.

In some embodiments, X ₁ is K and X ₁₀ is Q. In some embodiments, X ₁ is K and X ₁₀ is S. In some embodiments, X ₁ is K and X ₁₅ is E. In some embodiments, X ₁ is K and X ₁₅ is F. In some embodiments, X ₁ is K and X ₁₆ is L. In some embodiments, X ₁ is K and X ₁₆ is K. In some embodiments, X ₁ is K and X ₁₆ is k. In some embodiments, X ₁ is K and X ₁₇ is H. In some embodiments, X ₁ is K and X ₁₇ is V. In some embodiments, X ₁ is K and X ₁₇ is Q. In some embodiments, X ₁ is K and X ₃₅ is E. In some embodiments, X ₁ is K and X ₃₅ is N.

In some embodiments, X ₁ is k and X ₁₀ is Q. In some embodiments, X ₁ is k and X ₁₀ is S. In some embodiments, X ₁ is k and X ₁₅ is E. In some embodiments, X ₁ is k and X ₁₅ is F. In some embodiments, X ₁ is k and X ₁₆ is L. In some embodiments, X ₁ is k and X ₁₆ is K. In some embodiments, X ₁ is k and X ₁₆ is k. In some embodiments, X ₁ is k and X ₁₇ is H. In some embodiments, X ₁ is k and X ₁₇ is V. In some embodiments, X ₁ is k and X ₁₇ is Q. In some embodiments, X ₁ is k and X ₃₅ is E. In some embodiments, X ₁ is k and X ₃₅ is N.

In some embodiments, X ₁₀ is Q and X ₁₅ is E. In some embodiments, X ₁₀ is Q and X ₁₅ is F. In some embodiments, X ₁₀ is Q and X ₁₆ is L. In some embodiments, X ₁₀ is Q and X ₁₆ is K. In some embodiments, X ₁₀ is Q and X ₁₆ is k. In some embodiments, X ₁₀ is Q and X ₁₇ is H. In some embodiments, X ₁₀ is Q and X ₁₇ is V. In some embodiments, X ₁₀ is Q and X ₁₇ is Q. In some embodiments, X ₁₀ is Q and X ₃₅ is E. In some embodiments, X ₁₀ is Q and X ₃₅ is N.

In some embodiments, X ₁₀ is S and X ₁₅ is E. In some embodiments, X ₁₀ is S and X ₁₅ is F. In some embodiments, X ₁₀ is S and X ₁₆ is L. In some embodiments, X ₁₀ is S and X ₁₆ is K. In some embodiments, X ₁₀ is S and X ₁₆ is k. In some embodiments, X ₁₀ is S and X ₁₇ is H. In some embodiments, X ₁₀ is S and X ₁₇ is V. In some embodiments, X ₁₀ is S and X ₁₇ is Q. In some embodiments, X ₁₀ is S and X ₃₅ is E. In some embodiments, X ₁₀ is S and X ₃₅ is N.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₆ is K. In some embodiments, X ₁₅ is E and X ₁₇ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₇ is V. In some embodiments, X ₁₅ is E and X ₁₇ is Q. In some embodiments, X ₁₅ is E and X ₃₅ is E. In some embodiments, X ₁₅ is E and X ₃₅ is N.

In some embodiments, X ₁₅ is F and X ₁₆ is L. In some embodiments, X ₁₅ is F and X ₁₆ is K. In some embodiments, X ₁₅ is F and X ₁₆ is k. In some embodiments, X ₁₅ is F and X ₁₇ is H. In some embodiments, X ₁₅ is F and X ₁₇ is V. In some embodiments, X ₁₅ is F and X ₁₇ is Q. In some embodiments, X ₁₅ is F and X ₃₅ is E. In some embodiments, X ₁₅ is F and X ₃₅ is N.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₇ is V. In some embodiments, X ₁₆ is L and X ₁₇ is Q. In some embodiments, X ₁₆ is L and X ₃₅ is E. In some embodiments, X ₁₆ is L and X ₃₅ is N.

In some embodiments, X ₁₆ is K and X ₁₇ is H. In some embodiments, X ₁₆ is K and X ₁₇ is V. In some embodiments, X ₁₆ is K and X ₁₇ is Q. In some embodiments, X ₁₆ is K and X ₃₅ is E. In some embodiments, X ₁₆ is K and X ₃₅ is N.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₇ is V. In some embodiments, X ₁₆ is k and X ₁₇ is Q. In some embodiments, X ₁₆ is k and X ₃₅ is E. In some embodiments, X ₁₆ is k and X ₃₅ is N.

In some embodiments, X ₁₇ is H and X ₃₅ is E. In some embodiments, X ₁₇ is H and X ₃₅ is N.

In some embodiments, X ₁₇ is V and X ₃₅ is E. In some embodiments, X ₁₇ is V and X ₃₅ is N.

In some embodiments, X ₁₇ is Q and X ₃₅ is E. In some embodiments, X ₁₇ is Q and X ₃₅ is N.

In some embodiments, an isolated polypeptide of the invention comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus sequence of SEQ ID NO: 206: SCNTSTCATQRLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 206), wherein:

X ₁₅ represents E or F;

X ₁₆ represents L, K or k;

X ₁₇ represents H, V, or Q; and

X ₃₅ represents E or N;

each K is independently L-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

each k independently represents D-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer; and

wherein the two cysteine residues SCNTSTC (SEQ ID NO: 310) are optionally further linked by a disulfide bridge.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, an isolated polypeptide of the invention comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus sequence SEQ ID NO: 207: SCNTSTCATQRLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSETY-OH/NH ₂ ) (SEQ ID NO: 207), wherein:

X ₁₅ represents E or F;

X ₁₆ represents L, K, or k; and

X ₁₇ represents H, V or Q;

each K is independently L-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

each k independently represents D-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer; and

wherein the two cysteine residues of SC*NTSTC* are optionally additionally linked by a disulfide bridge.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 206 or SEQ ID NO: 207 include the following:

In some embodiments, carboxy-terminal amino acid 37 is Y-(NH ₂ ). In some embodiments, carboxy-terminal amino acid 37 is Y-(OH).

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₆ is K. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₇ is V. In some embodiments, X ₁₇ is E and X ₁₇ is Q. In some embodiments, X ₁₅ is E and X ₃₅ is E. In some embodiments, X ₁₅ is E and X ₃₅ is N.

In some embodiments, X ₁₅ is F and X ₁₆ is L. In some embodiments, X ₁₅ is F and X ₁₆ is K. In some embodiments, X ₁₅ is F and X ₁₆ is k. In some embodiments, X ₁₅ is F and X ₁₇ is H. In some embodiments, X ₁₅ is F and X ₁₇ is V. In some embodiments, X ₁₅ is F and X ₁₇ is Q. In some embodiments, X ₁₅ is F and X ₃₅ is E. In some embodiments, X ₁₅ is F and X ₃₅ is N.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₇ is V. In some embodiments, X ₁₆ is L and X ₁₇ is Q. In some embodiments, X ₁₆ is L and X ₃₅ is E. In some embodiments, X ₁₆ is L and X ₃₅ is N.

In some embodiments, X ₁₆ is K and X ₁₇ is H. In some embodiments, X ₁₆ is K and X ₁₇ is V. In some embodiments, X ₁₆ is K and X ₁₇ is Q. In some embodiments, X ₁₆ is K and X ₃₅ is E. In some embodiments, X ₁₆ is K and X ₃₅ is N.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₇ is V. In some embodiments, X ₁₆ is k and X ₁₇ is Q. In some embodiments, X ₁₆ is k and X ₃₅ is E. In some embodiments, X ₁₆ is k and X ₃₅ is N.

In some embodiments, X ₁₇ is H and X ₃₅ is E. In some embodiments, X ₁₇ is H and X ₃₅ is N.

In some embodiments, X ₁₇ is V and X ₃₅ is E. In some embodiments, X ₁₇ is V and X ₃₅ is N.

In some embodiments, X ₁₇ is Q and X ₃₅ is E. In some embodiments, X ₁₇ is Q and X ₃₅ is N.

In some embodiments, an isolated polypeptide of the invention comprises an amino acid sequence selected from the group consisting of:

SC*NTSTC*ATQRLANFkHKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 127), also referred to herein as compound A127;

SC*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 57), also referred to herein as compound A57;

SC*NTSTC*ATQRLANEKHKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 128), also referred to herein as compound A128;

SC*NTSTC*ATQRLANEkHKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 129), also referred to herein as compound A129; and

SC*NTSTC*ATQRLANFLVKSSNEFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 43), also referred to herein as compound A43.

In some embodiments, an isolated polypeptide of the invention comprises the amino acid sequence:

SC*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 57), also referred to herein as compound A57.

In some embodiments, an isolated polypeptide of the invention comprises the amino acid sequence:

SC*NTSTC*ATQRLANFkHKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 127), also referred to herein as compound A127.

In some embodiments, an isolated polypeptide of the invention comprises the amino acid sequence:

SC*NTSTC*ATQRLANEk*((γGlu) ₂ -CO(CH ₂ ) ₁₄ CH ₃ )HKSSNNFGPILPP TKVGSETY-NH ₂ (SEQ ID NO: 27), also referred to herein as compound A27.

In some embodiments, an isolated polypeptide of the invention comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus sequence of SEQ ID NO: 208: X ₁ CNTSTCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 208), wherein:

X ₁ represents K or k;

X ₁₀ represents Q or S;

X ₁₅ represents E or F;

X ₁₆ represents L, K or k;

X ₁₇ represents H, V, or Q; and

X ₃₅ represents E or N;

each K is independently L-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

each k independently represents D-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer; and

wherein the two cysteine residues X ₁ CNTSTC (SEQ ID NO: 318) are optionally further linked by a disulfide bridge.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, an isolated polypeptide of the invention comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus sequence of SEQ ID NO: 209: X ₁ CNTSTCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSETY-(OH/NH ₂ ) (SEQ ID NO: 209), wherein:

X ₁ represents K or k;

X ₁₀ represents Q or S;

X ₁₅ represents E or F;

X ₁₆ represents L, K, or k; and

X ₁₇ represents H, V or Q;

each K is independently L-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

each k independently represents D-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer; and

wherein the two cysteine residues X ₁ CNTSTC (SEQ ID NO: 318) are optionally further linked by a disulfide bridge.

In some embodiments, the specific amino acids represented by the consensus sequence of SEQ ID NO: 208 or SEQ ID NO: 209 include the following:

In some embodiments, carboxy-terminal amino acid 37 is Y-(NH ₂ ). In some embodiments, carboxy-terminal amino acid 37 is Y-(OH).

In some embodiments, X ₁ is K. In some embodiments, X ₁ is k.

In some embodiments, X ₁₀ is Q. In some embodiments, X ₁₀ is S.

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q.

In some embodiments, X ₁ is K and X ₁₀ is Q. In some embodiments, X ₁ is K and X ₁₀ is S. In some embodiments, X ₁ is K and X ₁₅ is E. In some embodiments, X ₁ is K and X ₁₅ is F. In some embodiments, X ₁ is K and X ₁₆ is L. In some embodiments, X ₁ is K and X ₁₆ is K. In some embodiments, X ₁ is K and X ₁₆ is k. In some embodiments, X ₁ is K and X ₁₇ is H. In some embodiments, X ₁ is K and X ₁₇ is V. In some embodiments, X ₁ is K and X ₁₇ is Q. In some embodiments, X ₁ is K and X ₃₅ is E. In some embodiments, X ₁ is K and X ₃₅ is N.

In some embodiments, X ₁ is k and X ₁₀ is Q. In some embodiments, X ₁ is k and X ₁₀ is S. In some embodiments, X ₁ is k and X ₁₅ is E. In some embodiments, X ₁ is k and X ₁₅ is F. In some embodiments, X ₁ is k and X ₁₆ is L. In some embodiments, X ₁ is k and X ₁₆ is K. In some embodiments, X ₁ is k and X ₁₆ is k. In some embodiments, X ₁ is k and X ₁₇ is H. In some embodiments, X ₁ is k and X ₁₇ is V. In some embodiments, X ₁ is k and X ₁₇ is Q. In some embodiments, X ₁ is k and X ₃₅ is E. In some embodiments, X ₁ is k and X ₃₅ is N.

In some embodiments, X ₁₀ is Q and X ₁₅ is E. In some embodiments, X ₁₀ is Q and X ₁₅ is F. In some embodiments, X ₁₀ is Q and X ₁₆ is L. In some embodiments, X ₁₀ is Q and X ₁₆ is K. In some embodiments, X ₁₀ is Q and X ₁₆ is k. In some embodiments, X ₁₀ is Q and X ₁₇ is H. In some embodiments, X ₁₀ is Q and X ₁₇ is V. In some embodiments, X ₁₀ is Q and X ₁₇ is Q. In some embodiments, X ₁₀ is Q and X ₃₅ is E. In some embodiments, X ₁₀ is Q and X ₃₅ is N.

In some embodiments, X ₁₀ is S and X ₁₅ is E. In some embodiments, X ₁₀ is S and X ₁₅ is F. In some embodiments, X ₁₀ is S and X ₁₆ is L. In some embodiments, X ₁₀ is S and X ₁₆ is K. In some embodiments, X ₁₀ is S and X ₁₆ is k. In some embodiments, X ₁₀ is S and X ₁₇ is H. In some embodiments, X ₁₀ is S and X ₁₇ is V. In some embodiments, X ₁₀ is S and X ₁₇ is Q. In some embodiments, X ₁₀ is S and X ₃₅ is E. In some embodiments, X ₁₀ is S and X ₃₅ is N.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₆ is K. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₇ is V. In some embodiments, X ₁₅ is E and X ₁₇ is Q. In some embodiments, X ₁₅ is E and X ₃₅ is E. In some embodiments, X ₁₅ is E and X ₃₅ is N.

In some embodiments, X ₁₅ is F and X ₁₆ is L. In some embodiments, X ₁₅ is F and X ₁₆ is K. In some embodiments, X ₁₅ is F and X ₁₆ is k. In some embodiments, X ₁₅ is F and X ₁₇ is H. In some embodiments, X ₁₅ is F and X ₁₇ is V. In some embodiments, X ₁₅ is F and X ₁₇ is Q. In some embodiments, X ₁₅ is F and X ₃₅ is E. In some embodiments, X ₁₅ is F and X ₃₅ is N.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₇ is V. In some embodiments, X ₁₆ is L and X ₁₇ is Q. In some embodiments, X ₁₆ is L and X ₃₅ is E. In some embodiments, X ₁₆ is L and X ₃₅ is N.

In some embodiments, X ₁₆ is K and X ₁₇ is H. In some embodiments, X ₁₆ is K and X ₁₇ is V. In some embodiments, X ₁₆ is K and X ₁₇ is Q. In some embodiments, X ₁₆ is K and X ₃₅ is E. In some embodiments, X ₁₆ is K and X ₃₅ is N.

In some embodiments, X ₁₆ is K and X ₁₇ is H. In some embodiments, X ₁₆ is K and X ₁₇ is V. In some embodiments, X ₁₆ is K and X ₁₇ is Q. In some embodiments, X ₁₆ is K and X ₃₅ is E. In some embodiments, X ₁₆ is K and X ₃₅ is N.

In some embodiments, X ₁₇ is H and X ₃₅ is E. In some embodiments, X ₁₇ is H and X ₃₅ is N.

In some embodiments, X ₁₇ is V and X ₃₅ is E. In some embodiments, X ₁₇ is V and X ₃₅ is N.

In some embodiments, X ₁₇ is Q and X ₃₅ is E. In some embodiments, X ₁₇ is Q and X ₃₅ is N.

In some embodiments, an isolated polypeptide of the invention comprises an amino acid sequence selected from the group consisting of:

KC*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 130), also referred to herein as compound A130; and

KC*NTSTC*ATQRLANFLQKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 131), also referred to herein as compound A131.

In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

5. Conjugation of a lipophilic substituent to any of the peptides, optionally using a spacer

In some embodiments, any of the disclosed polypeptides is optionally substituted with one or more lipophilic substituents, each optionally attached via a spacer, wherein "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 of SEQ ID NO: 1 to SEQ ID NO: 143 either comprises one or more lipophilic substituents, each optionally attached via a spacer, or may be altered or further modified by covalent attachment of one or more lipophilic substituents, each optionally attached via a spacer. In some embodiments, the lipophilic substituent may be attached to an amino group of the polypeptide (e.g., an ε-amino group from lysine residues) via the carboxyl group of the lipophilic substituent or, optionally, the amino group of a spacer, wherein the carboxyl group of the spacer forms an amide bond with the ε-amino group from lysine residues.

Lipophilic substitute

Conjugation of one or more "lipophilic substituents", each optionally attached via a spacer, to any of the disclosed peptides of the invention is intended to prolong the action of the polypeptide by facilitating binding to serum albumin and slowing the excretion of the conjugated polypeptide from the kidneys. As used herein, the term "lipophilic substituent" comprises a substituent containing from 4 to 40 carbon atoms, from 8 to 25 carbon atoms, from 12 to 22 carbon atoms, or from 6 to 20 carbon atoms. The lipophilic substituent may be attached to an amino group of the polypeptide (e.g., the ε-amino group of a lysine residue) via a carboxyl group of the lipophilic substituent, or, optionally, an amino group of a spacer, the carboxyl group of the spacer in turn forming an amide bond with the amino group of the amino acid (e.g., lysine) residue to which it is attached. In some embodiments, the polypeptide comprises three, two, or, preferably, one lipophilic substituent, each with or without an optional spacer, which is defined in more detail below.

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

In some embodiments, the polypeptide comprises three, two, or, preferably, one lipophilic substituent, each with or without an optional spacer. In some embodiments, the lipophilic substituent is a monovalent group of formula I:

Where

Z represents _-CH3 or _-CO2H ; and

m is from 4 to 24,

said lipophilic substituent forms an amide bond between the amino group (for example, the ε-amino group of lysine) of the disclosed polypeptide and the CO- group of the lipophilic substituent.

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, Z is -CO ₂ H and the lipophilic substituent has the formula -CO-(CH ₂ ) _m -CO ₂ H. In some embodiments, -CO-(CH ₂ ) _m -Z is selected from the group consisting of -CO-(CH ₂ ) ₄ -CO ₂ H, -CO-(CH ₂ ) ₅ -CO ₂ H, -CO-(CH ₂ ) ₆ -CO ₂ H, -CO-(CH ₂ ) ₇ -CO ₂ H, -CO-(CH ₂ ) ₈ -CO ₂ H, -CO-(CH ₂ ) ₉ -CO ₂ H, -CO-(CH ₂ ) ₁₀ -CO ₂ H, -CO-(CH ₂ ) ₁₁ -CO ₂ H, -CO-(CH ₂ ) ₁₂ -CO ₂ H, -CO-(CH ₂ ) ₁₃ -CO ₂ H, -CO-(CH ₂ ) ₁₄ -CO ₂ H, -CO-(CH ₂ ) ₁₅ -CO ₂ H, -CO-(CH ₂ ) ₁₆ -CO ₂ H, -CO-(CH ₂ ) ₁₇ -CO ₂ H, -CO-(CH ₂ ) ₁₈ -CO ₂ H, -CO-(CH ₂ ) ₁₉ -CO ₂ H, -CO-(CH ₂ ) ₂₀ -CO ₂ H.

In some embodiments, the lipophilic substituent is -CO-(CH ₂ ) ₁₈ -CO ₂ H.

In some embodiments, Z is -CH ₃ and the lipophilic substituent has the formula -CO-(CH ₂ ) _m -CH ₃ . In some embodiments, -CO-( _CH2 ) _m -Z is selected from the group consisting of -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 -CH ₃ , -CO-(CH ₂ ) ₁₇ -CH ₃ , -CO-(CH ₂ ) ₁₈ -CH ₃ , -CO-(CH ₂ ) ₁₉ -CH ₃ and -CO-(CH ₂ ) ₂₀ -CH ₃ .

Spacer

In some embodiments, the lipophilic substituent is attached to the parent peptide via a "spacer". In some embodiments, the document provides 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 NO: 1 to SEQ ID NO: 143, comprising a lipophilic substituent, wherein the lipophilic substituent is linked to the e-amino group of lysine via a spacer, wherein the spacer forms a bridge between the amino group of the disclosed polypeptide and the CO- group of the lipophilic substituent.

In some embodiments, the spacer comprises one or more amino acids, such as 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) (SEQ ID NO: 317), 2(Glu)-Gly, etc. In some embodiments, when the spacer comprises one or more amino acids, such as 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 the amino group of the spacer can form an amide bond with a carboxyl group of a lipophilic substituent.

In some embodiments, when the spacer comprises Glu or Asp, which further comprise a side chain on a terminal carboxylic acid, the terminal carboxyl group of the side chain of the Glu or Asp-containing spacer can form an amide bond with an amino group of the disclosed polypeptide, and the amino group of the Glu or Asp-containing spacer can form an amide bond with a carboxyl group of a lipophilic substituent, i.e., γGlu or βAsp. In some embodiments, the spacer is γGlu. In some embodiments, the spacer is 2(γGlu). In some embodiments, the spacer is 3(γGlu).

In some embodiments, the polypeptide comprises three, two, or, preferably, one lipophilic substituent, each with a spacer. In some embodiments, the lipophilic substituent and the spacer are a monovalent group of formula II:

Where

Y is selected from the group consisting of γGlu, Asp, Lys and Gly;

Z represents _-CH3 or _-CO2H ;

m is from 4 to 24; And

n is from 1 to 10.

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.

In some embodiments, the polypeptide comprises three, two, or, preferably, one lipophilic substituent, each with a spacer. In some embodiments, the lipophilic substituent and the spacer are a monovalent group of formula III:

Where

Z represents _-CH3 or _-CO2H ;

m is from 4 to 24; And

n is from 1 to 10.

In some embodiments, Z is _-CH3 . In some embodiments, Z is _-CO2H .

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, 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.

In some embodiments, the polypeptide comprises three, two, or, preferably, one lipophilic substituent, each with a spacer. In some embodiments, the lipophilic substituent and the spacer are a monovalent group of formula IV:

Where

Z represents _-CH3 or _-CO2H ;

m is from 4 to 24; And

n is from 1 to 10.

In some embodiments, (γGlu)n is selected from the group consisting of γGlu; 2(γGlu); 3(γGlu); 4(γGlu) (SEQ ID NO: 313) and 5(γGlu) (SEQ ID NO: 314). In some embodiments, -(γGlu)n-(Gly)- ("(γGlu)n-(Gly)" disclosed as SEQ ID NO: 312) is selected from the group consisting of 2(γGlu),Gly and 3(γGlu),Gly (SEQ ID NO: 315).

In some embodiments, the polypeptide comprises three, two, or, preferably, one lipophilic substituent, each with a spacer. In some embodiments, the lipophilic substituent and the spacer are a monovalent group of formula V:

Where

Z represents _-CH3 or _-CO2H ;

m is from 4 to 24; And

n is from 1 to 10.

In some embodiments, certain variables presented in Formula II, Formula III, Formula IV, or Formula V include the following:

In some embodiments, Z is _-CH3 . In some embodiments, Z is _-CO2H .

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, 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.

In some embodiments, n is 1 and Z is -CO ₂ H. In some embodiments, n is 1 and Z is -CH ₃ . In some embodiments, n is 2 and Z is -CO ₂ H. In some embodiments, n is 2 and Z is -CH ₃ . In some embodiments, n is 3 and Z is -CO ₂ H. In some embodiments, n is 3 and Z is -CH ₃ . In some embodiments, n is 4 and Z is -CO ₂ H. In some embodiments, n is 4 and Z is -CH ₃ . In some embodiments, n is 5 and Z is -CO ₂ H. In some embodiments, n is 5 and Z is -CH ₃ .

In some embodiments, n is 1, Z is -CO ₂ H, am is 14-20. In some embodiments, n is 1, Z is -CO ₂ H, am is 14. In some embodiments, n is 1, Z is -CO ₂ H, am is 16. In some embodiments, n is 1, Z is -CO ₂ H, am is 18.

In some embodiments, n is 1, Z is -CH ₃ , am is 14-20. In some embodiments, n is 1, Z is -CH ₃ , am is 14. In some embodiments, n is 1, Z is -CH ₃ , am is 16. In some embodiments, n is 1, Z is -CH ₃ , am is 18.

In some embodiments, n is 2, Z is _-CO2H , am is 14-20. In some embodiments, n is 2, Z is _-CO2H , am is 14. In some embodiments, n is 2, Z is -CO2H, am is _16. In some embodiments, n is 2, Z is _-CO2H , am is 18.

In some embodiments, n is 2, Z is -CH ₃ , am is 14-20. In some embodiments, n is 2, Z is -CH ₃ , am is 14. In some embodiments, n is 2, Z is -CH ₃ , am is 16. In some embodiments, n is 2, Z is -CH ₃ , am is 18.

In some embodiments, n is 3, Z is _-CO2H , am is 14-20. In some embodiments, n is 3, Z is _-CO2H , am is 14. In some embodiments, n is 3, Z is -CO2H, am is _16. In some embodiments, n is 3, Z is _-CO2H , am is 18.

In some embodiments, n is 3, Z is -CH ₃ , am is 14-20. In some embodiments, n is 3, Z is -CH ₃ , am is 14. In some embodiments, n is 3, Z is -CH ₃ , am is 16. In some embodiments, n is 3, Z is -CH ₃ , am is 18.

In some embodiments, n is 4, Z is _-CO2H , am is 14-20. In some embodiments, n is 4, Z is _-CO2H , am is 14. In some embodiments, n is 4, Z is -CO2H, am is _16. In some embodiments, n is 4, Z is _-CO2H , am is 18.

In some embodiments, n is 4, Z is -CH ₃ , am is 14-20. In some embodiments, n is 4, Z is -CH ₃ , am is 14. In some embodiments, n is 4, Z is -CH ₃ , am is 16. In some embodiments, n is 4, Z is -CH ₃ , am is 18.

In some embodiments, n is 5, Z is _-CO2H , am is 14-20. In some embodiments, n is 5, Z is _-CO2H , am is 14. In some embodiments, n is 5, Z is -CO2H, am is _16. In some embodiments, n is 5, Z is _-CO2H , am is 18.

In some embodiments, n is 5, Z is -CH ₃ , am is 14-20. In some embodiments, n is 5, Z is -CH ₃ , am is 14. In some embodiments, n is 5, Z is -CH ₃ , am is 16. In some embodiments, n is 5, Z is -CH ₃ , am is 18.

In some embodiments, the polypeptide comprises three, two, or, preferably, one lipophilic substituent, each with a spacer. In some embodiments, the lipophilic substituent and the spacer are a monovalent group of formula VI:

Where

Z represents _-CH3 or _-CO2H ;

m is from 4 to 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 (O( _CH2 ) ₂ ) _tOCH2NH ]-, t is from ₁ to 8;

r is from 1 to 8;

n1 is from 0 to 10; and

n2 is from 0 to 10.

In some embodiments, the polypeptide comprises three, two, or, preferably, one lipophilic substituent, each with a spacer. In some embodiments, the lipophilic substituent and the spacer are a monovalent group of formula VII:

Where

Z represents _-CH3 or _-CO2H ; m is from 4 to 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]-;

r is from 1 to 8;

n1 is from 0 to 10; and

n2 is from 0 to 10.

In some embodiments, -(Y1) _n1 -(dpeg)r-(Y2) _n2 - is selected from the group consisting of γGlu,dpeg,dpeg,γGlu; γGlu,dpeg,dpeg,2(γGlu); γGlu,dpeg,dpeg,3(γGlu); γGlu,dpeg,dpeg,4(γGlu); 2(γGlu),dpeg,dpeg,γGlu; and 2(γGlu),dpeg,γGlu.

In some embodiments, the polypeptide comprises three, two, or, preferably, one lipophilic substituent, each with a spacer. In some embodiments, the lipophilic substituent and the spacer are a monovalent group of formula VIII:

Where

Z represents _-CH3 or _-CO2H ; m is from 4 to 24;

Y2 is selected from the group consisting of γGlu, Asp and Gly;

V is -[ _COCH2 (O( _CH2 ) ₂ ) _tOCH2NH ]-, t is from ₁ to 8;

r is from 1 to 8; and

n2 is from 0 to 10.

In some embodiments, the polypeptide comprises three, two, or, preferably, one lipophilic substituent, each with a spacer. In some embodiments, the lipophilic substituent and the spacer are a monovalent group of formula IX:

Where

Z represents _-CH3 or _-CO2H ;

m is from 4 to 24;

dpeg is -[CO(CH ₂ )O(CH ₂ ) ₂ O(CH ₂ )NH]-;

Y2 is selected from the group consisting of γGlu, Asp and Gly;

r is from 1 to 8; and

n2 is from 0 to 10.

In some embodiments, -(dpeg) _r -(Y2) _n2 - is selected from the group consisting of dpeg,γGlu and dpeg,dpeg,γGlu.

In some embodiments, the polypeptide comprises three, two, or, preferably, one lipophilic substituent, each with a spacer. In some embodiments, the lipophilic substituent and the spacer are a monovalent group of formula X:

Where

Z represents _-CH3 or _-CO2H ;

m is from 4 to 24;

Y1 is selected from the group consisting of γGlu, Asp and Gly;

dpeg is -[CO(CH ₂ )O(CH ₂ ) ₂ O(CH ₂ )NH]-;

r is from 1 to 8; and

n1 is from 0 to 10.

In some embodiments, -(Y1) _n1 -(dpeg) _r - is 2(γGlu),dpeg.

In some embodiments of the present invention, the spacer comprises a bivalent group of formula XI:

Where

each R ₁ and R ₃ is hydrogen or C ₁ -C ₄ alkyl;

each R ₂ represents H or CO ₂ H;

p is 1, 2, 3, 4, 5, or 6;

q is 1, 2, or 3;

r is 0 or 1.

This spacer forms a bridge between the amino group of the exposed polypeptide and the CO group of the lipophilic substituent.

In some embodiments of the present invention, the spacer comprises a bivalent group of formula XII:

Where

each R ₃ is hydrogen or C ₁ -C ₄ alkyl;

q is 1, 2, or 3;

r is 0 or 1.

This spacer forms a bridge between the amino group of the exposed polypeptide and the CO group of the lipophilic substituent.

In some embodiments, certain variables presented in Formula XI or Formula XII include the following:

In some embodiments, each R ₁ is hydrogen. In some embodiments, each R ₃ is hydrogen. In some embodiments, each R ₁ and each R ₃ are hydrogen.

In some embodiments, at least one R ₂ is CO ₂ H. In some embodiments, one R ₂ is CO ₂ H.

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.

In some embodiments, q is 1. In some embodiments, q is 2. In some embodiments, q is 3.

In some embodiments, r is 0. In some embodiments, r is 1.

In some embodiments, the spacer is γ-glutamyl, i.e., -NH(CHCO ₂ H)(CH ₂ ) ₂ CO--. In some embodiments, the spacer is γ-aminobutanoyl, i.e., -NH(CH ₂ ) ₃ CO--. In some embodiments, the spacer is β-asparagyl, i.e., -NH(CHCO ₂ H)(CH ₂ )CO--. In some embodiments, the spacer is NH(CH ₂ ) ₂ CO--. In some embodiments, the spacer is glycyl. In some embodiments, the spacer is β-alanyl.

In some embodiments, the spacer is -NHCH(CO ₂ H)(CH ₂ ) ₂ CO-- [N(R ₃ )((CH ₂ ) ₂ O(CH ₂ ) ₂ O) _q (CH ₂ )CO-] _r . In some embodiments, the spacer is -NH(CH ₂ ) ₃ CO--[N(R ₃ )((CH ₂ ) ₂ O(CH ₂ ) ₂ O) _q (CH ₂ )CO-] _r . In some embodiments, the spacer is NHCH(CO ₂ H)(CH ₂ ) ₂ CO-NH((CH ₂ ) ₂ O(CH ₂ ) ₂ O) ₂ (CH ₂ )CO-. In some embodiments, the spacer is -NH(CH ₂ ) ₃ CO-NH((CH ₂ ) ₂ O(CH ₂ ) ₂ O) ₂ (CH ₂ )CO-. In some embodiments, the spacer is -NHCH(CO ₂ H)CH ₂ CO--[N(R ₃ )((CH ₂ ) ₂ O(CH ₂ ) ₂ O) _q (CH ₂ )CO-] _r . In some embodiments, the spacer is -NH(CH ₂ ) ₂ CO-- [N(R ₃ )((CH ₂ ) ₂ O(CH ₂ ) ₂ O) _q (CH ₂ )CO-] _r .

In some embodiments of the present invention, the spacer comprises a bivalent group of formula XIII:

Where

Y is selected from the group consisting of γGlu, Asp, Lys and Gly; n is from 1 to 10.

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.

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.

In some embodiments of the present invention, the spacer comprises a bivalent group of formula XIV:

Where

n is from 1 to 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.

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.

In some embodiments of the present invention, the spacer comprises a bivalent group of formula XV:

Where

n is from 1 to 10.

In some embodiments, (γGlu)n is selected from the group consisting of γGlu; 2(γGlu); 3(γGlu); 4(γGlu) (SEQ ID NO: 313) and 5(γGlu) (SEQ ID NO: 314). In some embodiments, -(γGlu)n-(Gly)- ("(γGlu)n-(Gly)" disclosed as SEQ ID NO: 312) is selected from the group consisting of 2(γGlu),Gly and 3(γGlu),Gly (SEQ ID NO: 315).

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.

In some embodiments of the present invention, the spacer comprises a bivalent group of formula XVI:

Where

n is from 1 to 10.

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.

In some embodiments of formula XIII, formula XIV, formula XV, or formula XVI, 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.

In some embodiments of the present invention, the spacer comprises a bivalent group of formula XVII:

Where

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 (O( _CH2 ) ₂ ) _tOCH2NH ]-, t is from ₁ to 8;

r is from 1 to 8;

n1 is from 0 to 10; and

n2 is from 0 to 10.

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.

In some embodiments of the present invention, the spacer comprises a bivalent group of formula XVIII:

Where

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]-;

r is from 1 to 8;

n1 is from 0 to 10; and

n2 is from 0 to 10.

In some embodiments, -(Y1) _n1 -(dpeg) _r -(Y2) _n2 - is selected from the group consisting of γGlu,dpeg,dpeg,γGlu; γGlu,dpeg,dpeg,2(γGlu); γGlu,dpeg,dpeg,3(γGlu); γGlu,dpeg,dpeg,4(γGlu); 2(γGlu),dpeg,dpeg,γGlu; and 2(γGlu),dpeg,γGlu.

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.

Accordingly, in some embodiments, the isolated polypeptide provided herein comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus

a sequence selected from the group consisting of SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208 and SEQ ID NO: 209, wherein the isolated peptide further comprises a lipophilic substituent, and optionally comprises a spacer.

In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 199, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent of formula I. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 199, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer selected from the group consisting of formula II, formula III, formula IV, formula V, formula VI, formula VII, formula VIII, formula GC, and formula X. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 199, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer of formula III. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 199, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer of formula VI. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 199 or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer of formula VII.

In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 200, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent of formula I. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 200, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer selected from the group consisting of formula II, formula III, formula IV, formula V, formula VI, formula VII, formula VIII, formula GC, and formula X. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 200, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer of formula III. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 200, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer of formula VI. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 200 or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer of formula VII.

In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 204, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent of formula I. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 204, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer selected from the group consisting of formula II, formula III, formula IV, formula V, formula VI, formula VII, formula VIII, formula GC, and formula X. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 204, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer of formula III. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 204, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer of formula VI. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 204 or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer of formula VII.

In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 206, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent of formula I. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 206, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer selected from the group consisting of formula II, formula III, formula IV, formula V, formula VI, formula VII, formula VIII, formula GC, and formula X. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 206, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer of formula III. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 206, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer of formula VI. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 206 or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer of formula VII.

In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 208, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent of formula I. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 208, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer selected from the group consisting of formula II, formula III, formula IV, formula V, formula VI, formula VII, formula VIII, formula GC, and formula X. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 208, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer of formula III. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 208, or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer of formula VI. In one embodiment, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 208 or a pharmaceutically acceptable salt thereof, further comprising a lipophilic substituent and a spacer of formula VII.

In some embodiments, an isolated polypeptide provided herein comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus sequence of SEQ ID NO: 200: X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 200), or a pharmaceutically acceptable salt thereof, wherein:

X ₁ is S, K, k, H, or I; X ₃ is N or S; X ₅ is S or A; X ₆ is T or S; X ₈ is A or K; X ₁₀ is Q or S; X ₁₂ is L or K; X ₁₃ is A, S, E or K; X ₁₄ is N, n, d, Y or Q; X ₁₅ is E, F, f, Y, I, k, K or Aib; X ₁₀ is k, K, L, Aib, N-MeL, or 1; X ₁₇ is H, V, Q, R, k, K or Aib; X ₁₈ is K, H, or R; X ₁₀ is S or Aib; X ₂₀ is S or Aib; X ₂₂ is N or E; X ₂₉ is P, R or K; X ₃₁ is k, K or N, and X ₃₅ is e, E or N; each K is independently L-lysine, optionally covalently linked to a lipophilic substituent, optionally via a spacer; each k is independently D-lysine, optionally covalently linked to a lipophilic substituent, optionally via a spacer; wherein the two cysteine residues X ₁ CX ₃ TX ₅ X ₆ C are optionally further linked by a disulfide bridge, and with the proviso that if X ₃₁ is N then X ₃₅ is E, or if X ₃₅ is N then X ₃₁ is K; and wherein the isolated peptide further comprises a lipophilic substituent of formula I:

Where

Z represents _-CH3 or _-CO2H ; and

m is from 4 to 24.

In some embodiments, an isolated polypeptide provided herein comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus sequence SEQ ID NO: 200:

X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁ 7X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 200), or a pharmaceutically acceptable salt thereof, wherein:

X ₁ is S, K, k, H, or I; X ₃ is N or S; X ₅ is S or A; X ₆ is T or S; X ₈ is A or K; X ₁₀ is Q or S; X ₁₂ is L or K; X ₁₃ is A, S, E or K; X ₁₄ is N, n, d, Y or Q; X ₁₅ is E, F, f, Y, I, k, K or Aib; X ₁₆ is k, K, L, Aib, N-MeL, or 1; X ₁₇ is H, V, Q, R, k, K or Aib; X ₁₈ is K, H, or R; X ₁₉ is S or Aib; X ₂₀ is S or Aib; X ₂₂ is N or E; X ₂₉ is P, R or K; X ₃₁ is k, K or N, and X ₃₅ is e, E or N; each K is independently L-lysine, optionally covalently linked to a lipophilic substituent, optionally via a spacer; each k is independently D-lysine, optionally covalently linked to a lipophilic substituent, optionally via a spacer; wherein the two cysteine residues X ₁ CX ₃ TX ₅ X ₆ C are optionally further linked by a disulfide bridge, and with the proviso that if X ₃₁ is N then X ₃₅ is E, or if X ₃₅ is N then X ₃₁ is K; and wherein the isolated peptide further comprises a lipophilic substituent and a spacer of formula III:

Where

Z represents _-CH3 or _-CO2H ;

m is from 4 to 24; And

n is from 1 to 10.

In some embodiments, an isolated polypeptide provided herein comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus sequence SEQ ID NO: 200:

X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 200), or a pharmaceutically acceptable salt thereof, wherein:

X ₁ is S, K, k, H, or I; X ₃ is N or S; X ₅ is S or A; X ₆ is T or S; Xs is A or K; X ₁₀ is Q or S; X ₁₂ is L or K; X ₁₃ is A, S, E or K; X ₁₄ is N, n, d, Y or Q; X ₁₅ is E, F, f, Y, I, k, K or Aib; X ₁₆ is k, K, L, Aib, N-MeL, or 1; X ₁₇ is H, V, Q, R, k, K or Aib; X ₁₈ is K, H, or R; X ₁₉ is S or Aib; X ₂₀ is S or Aib; X ₂₂ is N or E; X ₂₉ is P, R or K; X ₃₁ is k, K or N, and X ₃₅ is e, E or N; each K is independently L-lysine, optionally covalently linked to a lipophilic substituent, optionally via a spacer; each k is independently D-lysine, optionally covalently linked to a lipophilic substituent, optionally via a spacer; wherein the two cysteine residues X ₁ CX ₃ TX ₅ X ₆ C are optionally further linked by a disulfide bridge, and with the proviso that if X ₃₁ is N then X ₃₅ is E, or if X ₃₅ is N then X ₃₁ is K; and wherein the isolated peptide further comprises a lipophilic substituent and a spacer of formula VI:

Where

Z represents _-CH3 or _-CO2H ;

m is from 4 to 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 -[COCH ₂ (O(CH ₂ ) ₂ ) _t OCH ₂ NH]-, t is from 1 to 8;

r is from 1 to 8;

n1 is from 0 to 10; and

n2 is from 0 to 10.

In some embodiments, an isolated polypeptide provided herein comprises an amino acid sequence, or a pharmaceutically acceptable salt thereof, selected from the group consisting of amino acid sequences represented by the consensus sequence SEQ ID NO: 200:

X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁ 7X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 200), or a pharmaceutically acceptable salt thereof, wherein:

X ₁ is S, K, k, H, or I; X ₃ is N or S; X ₅ is S or A; X ₆ is T or S; Xs is A or K; X ₁₀ is Q or S; X ₁₂ is L or K; X ₁₀ is A, S, E or K; X ₁₀ is N, n, d, Y or Q; X ₁₀ is E, F, f, Y, I, k, K or Aib; X ₁₀ is k, K, L, Aib, N-MeL, or 1; X ₁₀ is H, V, Q, R, k, K or Aib; X ₁₀ is K, H, or R; X ₁₀ is S or Aib; X ₂₀ is S or Aib; X ₂₂ is N or E; X ₂₉ is P, R or K; X ₃₁ is k, K or N, and X ₃₅ is e, E or N; each K is independently L-lysine, optionally covalently linked to a lipophilic substituent, optionally via a spacer; each k is independently D-lysine, optionally covalently linked to a lipophilic substituent, optionally via a spacer; wherein the two cysteine residues X ₁ CX ₃ TX ₅ X6C are optionally further linked by a disulfide bridge, and with the proviso that if X ₃₁ is N then X ₃₅ is E, or if X ₃₅ is N then X ₃₁ is K; and wherein the isolated peptide further comprises a lipophilic substituent and a spacer of formula VII:

Where

Z represents _-CH3 or _-CO2H ;

m is from 4 to 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]-;

r is from 1 to 8;

n1 is from 0 to 10; and

n2 is from 0 to 10.

In some embodiments, the isolated polypeptide comprises a lipophilic substituent at position _X1 , _X15 , or _X16 .

In some embodiments, an isolated polypeptide comprising a lipophilic substituent at position _X1 , _X15 , or _X16 is conjugated via a lysine at that position.

In some embodiments, the isolated polypeptide comprises a lipophilic substituent at position _X1

In some embodiments, the isolated polypeptide comprises a lipophilic substituent at position _X15 .

In some embodiments, the isolated polypeptide comprises a lipophilic substituent at position _X16 .

In some embodiments, an isolated polypeptide comprising a lipophilic substituent at position or _X1 is conjugated via a lysine at that position.

In some embodiments, an isolated polypeptide comprising a lipophilic substituent at position or _X15 is conjugated via a lysine at that position.

In some embodiments, an isolated polypeptide comprising a lipophilic substituent at position or _X10 is conjugated via a lysine at that position.

In some embodiments, an isolated polypeptide of the invention comprises the amino acid sequence:

SC*NTSTC*ATQRLANEk*((γGlu) ₂ -CO(CH ₂ ) ₁₄ CH ₃ )HKSSNNFGPILPP TKVGSETY-NH ₂ (SEQ ID NO: 27), also referred to herein as compound A27.

In some embodiments, an isolated polypeptide of the invention comprises the amino acid sequence:

K*((γGlu) _{2 (} CO( _CH2 ) _18CO2H ))C*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-( _NH2 ) (SEQ ID NO: 64), also referred to herein as compound A64.

In some embodiments, an isolated polypeptide of the invention comprises the amino acid sequence:

K*((γGlu) _{2 (} CO( _CH2 ) _16CO2H ))C*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-( _NH2 ) (SEQ ID NO: 65), also referred to herein as compound A65.

In some embodiments, an isolated polypeptide of the invention comprises the amino acid sequence: K*(γGlu-CO(CH ₂ ) ₁₆ CO ₂ H)C*NTSTC*ATSRLANFLQKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 109), also referred to herein as compound A109.

6. Illustrative Compounds: Amylin Analog Polypeptides

In some embodiments, an isolated polypeptide of the invention comprises an amino acid sequence selected from the group consisting of the following peptides listed in Table 3:

Notes:

• each K* is independently L-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

• each k* is independently D-lysine optionally covalently linked to a lipophilic substituent, optionally through a spacer;

• two cysteine residues (C*) in positions 2 and 7 are optionally additionally linked by a disulfide bridge;

• in the context of this document, (γGlu) ₂ and 2(γGlu) both mean -(γGlu)-(γGlu)-; (γClu) ₃ and 3(γGlu) both mean -(γGlu)-(γGlu)-(γGlu)-; etc.; and

• where a variable occurs more than one time in a given formula, each occurrence of that variable is independently determined. For example, for the group -(Y) _3- , where Y can be γGlu, Asp, Lys, or Gly, each Y is independently selected to be one of four amino acids. Accordingly, by way of non-limiting example, -(Y) _3- can be -(γGlu)-(γGlu)-(γGlu)-, -(γGlu)-(Asp)-(γGlu)-, -(Gly)-(Asp)-(γGlu)-, or -(Gly)-(γGlu)-(γGlu)-.

In some embodiments, the invention provides a compound listed in Table 3 above, or a pharmaceutically acceptable salt 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. In one embodiment, the compound is A27. In some embodiments, the compound is the acetate salt of Compound A27. In some embodiments, the compound is the TFA salt of Compound A27. In some embodiments, the compound is the HCl salt of Compound A27. In one embodiment, the compound is A57. In some embodiments, the compound is the acetate salt of Compound A57. In some embodiments, the compound is the TFA salt of Compound A57. In some embodiments, the compound is the HCl salt of Compound A57. In one embodiment, the compound is A64. In some embodiments, the compound is the acetate salt of Compound A64. In some embodiments, the compound is a TFA salt of compound A64. In some embodiments, the compound is a HCl salt of compound A64.

7. Intermediate products of polypeptides

In certain embodiments, the present invention also provides synthetic intermediates of isolated polypeptides that are amylin analogs. In some embodiments, an intermediate of a polypeptide of the invention is an isolated polypeptide comprising an amino acid sequence selected from the group consisting of amino acid sequences represented by the consensus sequence of SEQ ID NO: 210:

X ₁ CX ₃ TX ₅ X ₆ CX ₈ TX ₁₀ RX ₁₂ X ₁₃ X ₁₄ X ₁₅ X ₁₆ X ₁₇ X ₁₈ X ₁₉ X ₂₀ NX ₂₂ FGPILPX ₂₉ TX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 210), where:

X ₁ represents S, K, k, H, or I;

X ₃ represents N or S;

X ₅ represents S or A;

X ₆ represents T or S;

X ₈ represents A or K;

X ₁₀ represents Q or S;

X ₁₂ represents L or K;

X ₁₃ represents A, S, E or K;

X ₁₄ represents N, n, d, Y or Q;

X ₁₅ is E, F, f, Y, I, k, K, or α-aminoisobutyric acid (Aib);

X ₁₆ represents k, K, L, Aib, N-methylleucine (N-MeL), or l;

X ₁₇ represents H, V, Q, R, k, K or Aib;

X ₁₈ represents K, H, or R;

X ₁₉ represents S or Aib;

X ₂₀ represents S or Aib;

X ₂₂ represents N or E;

X ₂₉ represents P, R or K;

X ₃₁ represents k, K, or N; and

X ₃₅ represents e, E, or N;

each K independently represents L-lysine optionally covalently linked to a protecting group or a spacer optionally linked to a protecting group;

each k independently represents D-lysine optionally covalently linked to a protecting group or a spacer optionally linked to a protecting group;

wherein two cysteine residues X ₁ CX ₃ TX ₅ X ₆ C are optionally additionally linked by a disulfide bridge;

provided that if X ₃₁ is N then X ₃₅ is E, or if X ₃₅ is N then X ₃₁ is K.

In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, X ₃₁ is K and X ₃₅ is E.

In some embodiments, the specific amino acids represented by the consensus sequence SEQ ID NO: 210 include the following:

In some embodiments, carboxy-terminal amino acid 37 is Y-(NH ₂ ). In some embodiments, carboxy-terminal amino acid 37 is Y-(OH).

In some embodiments, X ₁ is S. In some embodiments, X ₁ is K. In some embodiments, X ₁ is k. In some embodiments, X ₁ is H. In some embodiments, X ₁ is I.

In some embodiments, X ₃ is N. In some embodiments, X ₃ is S.

In some embodiments, X ₅ is S. In some embodiments, X ₅ is A.

In some embodiments, X ₆ is T. In some embodiments, X ₆ is S.

In some embodiments, X ₈ is A. In some embodiments, X ₈ is K.

In some embodiments, X ₁₀ is Q. In some embodiments, X ₁₀ is S.

In some embodiments, X ₁₂ is L. In some embodiments, X ₁₂ is K.

In some embodiments, X ₁₃ is A. In some embodiments, X ₁₃ is S. In some embodiments, X ₁₃ is E. In some embodiments, X ₁₃ is K.

In some embodiments, X ₁₃ is N. In some embodiments, X ₁₄ is n. In some embodiments, X ₁₄ is d. In some embodiments, X ₁₄ is Y. In some embodiments, X ₁₄ is Q.

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F. In some embodiments, X ₁₅ is f. In some embodiments, X ₁₅ is Y. In some embodiments, X ₁₅ is I. In some embodiments, X ₁₅ is K. In some embodiments, X ₁₅ is k. In some embodiments, X ₁₅ is Aib.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is I. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k. In some embodiments, X ₁₆ is Aib. In some embodiments, X ₁₆ is N-MeL.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q. In some embodiments, X ₁₇ is R. In some embodiments, X ₁₇ is K. In some embodiments, X ₁₇ is k. In some embodiments, X ₁₇ is Aib.

In some embodiments, X ₁₈ is K. In some embodiments, X ₁₈ is H. In some embodiments, X ₁₈ is R.

In some embodiments, X ₁₉ is S. In some embodiments, X ₁₉ is Aib.

In some embodiments, X ₂₀ is S. In some embodiments, X ₂₀ is Aib.

In some embodiments, X ₂₂ is N. In some embodiments, X ₂₂ is E.

In some embodiments, X ₂₉ is P. In some embodiments, X ₂₉ is R. In some embodiments, X ₂₉ is K.

In some embodiments, X ₃₁ is k. In some embodiments, X ₃₁ is K. In some embodiments, X ₃₁ is N.

In some embodiments, X ₃₅ is e. In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the specific amino acids represented by the consensus sequence SEQ ID NO: 210 include the following:

In some embodiments, X ₁ is S and X ₅ is S. In some embodiments, X ₁ is S and X ₁₀ is Q. In some embodiments, X ₁ is S and X ₁₅ is E. In some embodiments, X ₁ is S and X ₁₆ is L. In some embodiments, X ₁ is S and X ₁₆ is k. In some embodiments, X ₁ is S and X ₁₇ is H. In some embodiments, X ₁ is S and X ₁₈ is K. In some embodiments, X ₁ is S and X ₃₁ is K. In some embodiments, X ₁ is S and X ₃₅ is E.

In some embodiments, X ₁ is K and X ₅ is S. In some embodiments, X ₁ is K and X ₁₀ is Q. In some embodiments, X ₁ is K and X ₁₅ is E. In some embodiments, X ₁ is K and X ₁₆ is L. In some embodiments, X ₁ is K and X ₁₆ is k. In some embodiments, X ₁ is K and X ₁₇ is H. In some embodiments, X ₁ is K and X ₁₈ is K. In some embodiments, X ₁ is K and X ₃₁ is K. In some embodiments, X ₁ is k and X ₃₅ is E.

In some embodiments, X ₁ is k and X ₅ is S. In some embodiments, X ₁ is k and X ₁₀ is Q. In some embodiments, X ₁ is k and X ₁₅ is E. In some embodiments, X ₁ is k and X ₁₆ is L. In some embodiments, X ₁ is k and X ₁₆ is k. In some embodiments, X ₁ is k and X ₁₇ is H. In some embodiments, X ₁ is k and X ₁₈ is K. In some embodiments, X ₁ is k and X ₃₁ is K. In some embodiments, X ₁ is k and X ₃₅ is E.

In some embodiments, X ₅ is S and X ₁₅ is E. In some embodiments, X ₅ is S and X ₁₀ is Q. In some embodiments, X ₅ is S and X ₁₆ is L. In some embodiments, X ₅ is S and X ₁₆ is k. In some embodiments, X ₅ is S and X ₁₇ is H. In some embodiments, X ₅ is S and X ₁₈ is K. In some embodiments, X ₅ is S and X ₃₁ is K. In some embodiments, X ₅ is S and X ₃₅ is E.

In some embodiments, X ₁₀ is Q and X ₁₅ is E. In some embodiments, X ₁₀ is Q and X ₁₆ is L. In some embodiments, X ₁₀ is Q and X ₁₆ is k. In some embodiments, X ₁₀ is Q and X ₁₇ is H. In some embodiments, X ₁₀ is Q and X ₁₈ is K. In some embodiments, X ₁₀ is Q and X ₃₁ is K. In some embodiments, X ₁₀ is Q and X ₃₅ is E.

In some embodiments, X ₁₅ is E and X ₁₀ is L. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₆ is K. In some embodiments, X ₁₅ is E and X ₃₁ is K. In some embodiments, X ₁₅ is E and X ₃₅ is E.

In some embodiments, X ₁₀ is L and X ₁₇ is H. In some embodiments, X ₁₀ is L and X ₁₈ is K. In some embodiments, X ₁₀ is L and X ₃₁ is K. In some embodiments, X ₁₆ is L and X ₃₅ is E.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₈ is K. In some embodiments, X 16 is k and X 31 is K. In some embodiments, X ₁₆ is k and X ₃₁ is K. In some embodiments, X ₁₆ is k and X ₃₅ is E.

In some embodiments, X ₁₇ is H and X ₁₈ is K. In some embodiments, X ₁₇ is H and X ₃₁ is K. In some embodiments, X ₁₇ is H and X ₃₅ is E.

In some embodiments, X ₁₈ is K and X ₃₁ is K. In some embodiments, X ₁₈ is K and X ₃₅ is E.

In some embodiments, X ₃₁ is K and X ₃₅ is E.

In some embodiments, the specific amino acids represented by the consensus sequence SEQ ID NO: 210 include the following:

In some embodiments, X ₁ is S, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is S, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is S, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is S, and X ₃₅ is E.

In some embodiments, X ₁ is K, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is K, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is K, X ₅ is S, and X _1s is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is K, X ₅ is S, and X ₃₅ is E.

In some embodiments, X ₁ is k, X ₅ is S, and X ₁₅ is E. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₆ is L. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₆ is k. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₇ is H. In some embodiments, X ₁ is k, X ₅ is S, and X ₁₈ is K. In some embodiments, X ₁ is k, X ₅ is S, and X ₃₁ is K. In some embodiments, X ₁ is k, X ₅ is S, and X ₃₅ is E.

In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₆ is L. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₆ is k. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₈ is H. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₁₈ is K. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₃₁ is K. In some embodiments, X ₅ is S, X ₁₅ is E, and X ₃₅ is E.

In some embodiments, X ₁₀ is Q, X ₁₅ is E, and X ₁₆ is L. In some embodiments, X ₁₀ is Q, X ₁₆ is k, and X ₁₇ is H. In some embodiments, X ₁₀ is Q, X ₁₈ is K, and X ₃₁ is K. In some embodiments, X ₁₀ is Q, X ₃₁ is K, and X ₃₅ is E.

In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₆ is L, and X ₃₅ is E.

In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₁₇ is H. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₁₈ is K. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₁ is K. In some embodiments, X ₁₅ is E, X ₁₆ is k, and X ₃₅ is E.

In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is L, X ₁₇ is H, and X ₃₅ is E.

In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₁₈ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₁ is K. In some embodiments, X ₁₆ is k, X ₁₇ is H, and X ₃₅ is E.

In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₁ is K. In some embodiments, X ₁₇ is H, X ₁₈ is K, and X ₃₅ is E.

In some embodiments, X ₁₈ is K, X ₃₁ is K, and X ₃₅ is E.

In some embodiments, the intermediate product of the polypeptide of the invention is an isolated polypeptide comprising an amino acid sequence selected from the group consisting of amino acid sequences represented by the consensus sequence SEQ ID NO: 211:

X ₁ CNTSTCAiX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 211), where:

X ₁ represents S, K or k;

X ₁₀ represents Q or S;

X ₁₅ represents E or F;

X ₁₆ represents L, K or k;

X ₁₇ represents H, V, or Q; and

X ₃₅ represents E or N;

each K independently represents L-lysine optionally covalently linked to a protecting group or a spacer optionally linked to a protecting group;

each k is independently D-lysine optionally covalently linked to a protecting group or a spacer optionally linked to a protecting group; and

wherein the two cysteine residues X ₁ CNTSTC (SEQ ID NO: 309) are optionally further linked by a disulfide bridge.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the specific amino acids represented by the consensus sequence SEQ ID NO: 211 include the following:

In some embodiments, carboxy-terminal amino acid 37 is Y-(NH ₂ ). In some embodiments, carboxy-terminal amino acid 37 is Y-(OH).

In some embodiments, X ₁ is S. In some embodiments, X ₁ is K. In some embodiments, X ₁ is k.

In some embodiments, X ₁₀ is Q. In some embodiments, X ₁₀ is S.

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₀ is K. In some embodiments, X ₁₀ is k.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the specific amino acids represented by the consensus sequence SEQ ID NO: 211 include the following:

In some embodiments, X ₁ is S and X ₁₀ is Q. In some embodiments, X ₁ is S and X ₁₀ is S. In some embodiments, X ₁ is S and X ₁₅ is E. In some embodiments, X ₁ is S and X ₁₅ is F. In some embodiments, X ₁ is S and X ₁₆ is L. In some embodiments, X ₁ is S and X ₁₆ is K. In some embodiments, X ₁ is S and X ₁₆ is k. In some embodiments, X ₁ is S and X ₁₇ is H. In some embodiments, X ₁ is S and X ₁₇ is V. In some embodiments, X ₁ is S and X ₁₇ is Q. In some embodiments, X ₁ is S and X ₃₅ is E. In some embodiments, X ₁ is S and X ₃₅ is N.

In some embodiments, X ₁ is K and X ₁₀ is Q. In some embodiments, X ₁ is K and X ₁₀ is S. In some embodiments, X ₁ is K and X ₁₅ is E. In some embodiments, X ₁ is K and X ₁₅ is F. In some embodiments, X ₁ is K and X ₁₆ is L. In some embodiments, X ₁ is K and X ₁₆ is K. In some embodiments, X ₁ is K and X ₁₆ is k. In some embodiments, X ₁ is K and X ₁₇ is H. In some embodiments, X ₁ is K and X ₁₇ is V. In some embodiments, X ₁ is K and X ₁₇ is Q. In some embodiments, X ₁ is K and X ₃₅ is E. In some embodiments, X ₁ is K and X ₃₅ is N.

In some embodiments, X ₁ is k and X ₁₀ is Q. In some embodiments, X ₁ is k and X ₁₀ is S. In some embodiments, X ₁ is k and X ₁₅ is E. In some embodiments, X ₁ is k and X ₁₅ is F. In some embodiments, X ₁ is k and X ₁₆ is L. In some embodiments, X ₁ is k and X ₁₆ is K. In some embodiments, X ₁ is k and X ₁₆ is k. In some embodiments, X ₁ is k and X ₁₇ is H. In some embodiments, X ₁ is k and X ₁₇ is V. In some embodiments, X ₁ is k and X ₁₇ is Q. In some embodiments, X ₁ is k and X ₃₅ is E. In some embodiments, X ₁ is k and X ₃₅ is N.

In some embodiments, X ₁₀ is Q and X ₁₅ is E. In some embodiments, X ₁₀ is Q and X ₁₅ is F. In some embodiments, X ₁₀ is Q and X ₁₆ is L. In some embodiments, X ₁₀ is Q and X ₁₆ is K. In some embodiments, X ₁₀ is Q and X ₁₆ is k. In some embodiments, X ₁₀ is Q and X ₁₇ is H. In some embodiments, X ₁₀ is Q and X ₁₇ is V. In some embodiments, X ₁₀ is Q and X ₁₇ is Q. In some embodiments, X ₁₀ is Q and X ₃₅ is E. In some embodiments, X ₁₀ is Q and X ₃₅ is N.

In some embodiments, X ₁₀ is S and X ₁₅ is E. In some embodiments, X ₁₀ is S and X ₁₅ is F. In some embodiments, X ₁₀ is S and X ₁₆ is L. In some embodiments, X ₁₀ is S and X ₁₆ is K. In some embodiments, X ₁₀ is S and X ₁₀ is k. In some embodiments, X ₁₀ is S and X ₁₇ is H. In some embodiments, X ₁₀ is S and X ₁₇ is V. In some embodiments, X ₁₀ is S and X ₁₇ is Q. In some embodiments, X ₁₀ is S and X ₃₅ is E. In some embodiments, X ₁₀ is S and X ₃₅ is N.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₀ is K. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₇ is V. In some embodiments, X ₁₅ is E and X ₁₇ is Q. In some embodiments, X ₁₅ is E and X ₃₅ is E. In some embodiments, X ₁₅ is E and X ₃₅ is N.

In some embodiments, X ₁₅ is F and X ₁₆ is L. In some embodiments, X ₁₅ is F and X ₁₆ is K. In some embodiments, X ₁₅ is F and X ₁₆ is k. In some embodiments, X ₁₅ is F and X ₁₇ is H. In some embodiments, X ₁₅ is F and X ₁₇ is V. In some embodiments, X ₁₅ is F and X ₁₇ is Q. In some embodiments, X ₁₅ is F and X ₃₅ is E. In some embodiments, X ₁₅ is F and X ₃₅ is N.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₇ is V. In some embodiments, X ₁₆ is L and X ₁₇ is Q. In some embodiments, X ₁₆ is L and X ₃₅ is E. In some embodiments, X ₁₆ is L and X ₃₅ is N.

In some embodiments, X ₁₆ is K and X ₁₇ is H. In some embodiments, X ₁₆ is K and X ₁₇ is V. In some embodiments, X ₁₆ is K and X ₁₇ is Q. In some embodiments, X ₁₆ is K and X ₃₅ is E. In some embodiments, X ₁₀ is K and X ₃₅ is N.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₇ is V. In some embodiments, X ₁₆ is k and X ₁₇ is Q. In some embodiments, X ₁₆ is k and X ₃₅ is E. In some embodiments, X ₁₆ is k and X ₃₅ is N.

In some embodiments, X ₁₇ is H and X ₃₅ is E. In some embodiments, X ₁₇ is H and X ₃₅ is N.

In some embodiments, X ₁₇ is V and X ₃₅ is E. In some embodiments, X ₁₇ is V and X ₃₅ is N.

In some embodiments, X ₁₇ is Q and X ₃₅ is E. In some embodiments, X ₁₇ is Q and X ₃₅ is N.

In some embodiments, the intermediate product of the polypeptide of the invention is an isolated polypeptide comprising an amino acid sequence selected from the group consisting of amino acid sequences represented by the consensus sequence SEQ ID NO: 212:

SCNTSTCATQRLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 212), where:

X ₁₅ represents E or F;

X ₁₆ represents L, K or k;

X ₁₇ represents H, V, or Q; and

X ₃₅ represents E or N;

each K independently represents L-lysine optionally covalently linked to a protecting group or a spacer optionally linked to a protecting group;

each k is independently D-lysine optionally covalently linked to a protecting group or a spacer optionally linked to a protecting group; and

wherein the two cysteine residues SCNTSTC (SEQ ID NO: 310) are optionally further linked by a disulfide bridge.

In some embodiments, X ₃₅ is E. In some embodiments, X ₃₅ is N.

In some embodiments, the specific amino acids represented by the consensus sequence SEQ ID NO: 212 include the following:

In some embodiments, carboxy-terminal amino acid 37 is Y-(NH ₂ ). In some embodiments, carboxy-terminal amino acid 37 is Y-(OH).

In some embodiments, X ₁₅ is E. In some embodiments, X ₁₅ is F.

In some embodiments, X ₁₆ is L. In some embodiments, X ₁₆ is K. In some embodiments, X ₁₆ is k.

In some embodiments, X ₁₇ is H. In some embodiments, X ₁₇ is V. In some embodiments, X ₁₇ is Q.

In some embodiments, X ₁₅ is E and X ₁₆ is L. In some embodiments, X ₁₅ is E and X ₁₆ is K. In some embodiments, X ₁₅ is E and X ₁₆ is k. In some embodiments, X ₁₅ is E and X ₁₇ is H. In some embodiments, X ₁₅ is E and X ₁₇ is V. In some embodiments, X ₁₅ is E and X ₁₇ is Q. In some embodiments, X ₁₅ is E and X ₃₅ is E. In some embodiments, X ₁₅ is E and X ₃₅ is N.

In some embodiments, X ₁₅ is F and X ₁₆ is L. In some embodiments, X ₁₅ is F and X ₁₆ is K. In some embodiments, X ₁₅ is F and X ₁₆ is k. In some embodiments, X ₁₅ is F and X ₁₇ is H. In some embodiments, X ₁₅ is F and X ₁₇ is V. In some embodiments, X ₁₅ is F and X ₁₇ is Q. In some embodiments, X ₁₅ is F and X ₃₅ is E. In some embodiments, X ₁₅ is F and X ₃₅ is N.

In some embodiments, X ₁₆ is L and X ₁₇ is H. In some embodiments, X ₁₆ is L and X ₁₇ is V. In some embodiments, X ₁₆ is L and X ₁₇ is Q. In some embodiments, X ₁₆ is L and X ₃₅ is E. In some embodiments, X ₁₆ is L and X ₃₅ is N.

In some embodiments, X ₁₆ is K and X ₁₇ is H. In some embodiments, X ₁₆ is K and X ₁₇ is V. In some embodiments, X ₁₆ is K and X ₁₇ is Q. In some embodiments, X ₁₆ is K and X ₃₅ is E. In some embodiments, X ₁₆ is K and X ₃₅ is N.

In some embodiments, X ₁₆ is k and X ₁₇ is H. In some embodiments, X ₁₆ is k and X ₁₇ is V. In some embodiments, X ₁₆ is k and X ₁₇ is Q. In some embodiments, X ₁₆ is k and X ₃₅ is E. In some embodiments, X ₁₆ is k and X ₃₅ is N.

In some embodiments, X ₁₇ is H and X ₃₅ is E. In some embodiments, X ₁₇ is H and X ₃₅ is N.

In some embodiments, X ₁₇ is V and X ₃₅ is E. In some embodiments, X ₁₇ is V and X ₃₅ is N.

In some embodiments, X ₁₇ is Q and X ₃₅ is E. In some embodiments, X ₁₇ is Q and X ₃₅ is N.

In some embodiments, an isolated peptide comprising the amino acid sequence of SEQ ID NO: 210, SEQ ID NO: 211, or SEQ ID NO: 212 further comprises a protecting group or a spacer optionally linked to a protecting group. In some embodiments, the isolated peptide further comprises a protecting group. In some embodiments, the isolated peptide further comprises a spacer. In some embodiments, the isolated peptide further comprises a spacer linked to a protecting group.

In some embodiments, the isolated polypeptide comprises a protecting group or spacer, optionally linked to the protecting group at position _X1 , _X15 , or _X16 .

In some embodiments, an isolated polypeptide comprising a protecting group or spacer optionally linked to a protecting group at position _X1 , _X15 , or _X16 is conjugated via a lysine at that position.

In some embodiments, the isolated polypeptide comprises a protecting group or spacer, optionally linked to the protecting group at position _X1 .

In some embodiments, the isolated polypeptide comprises a protecting group or spacer, optionally linked to the protecting group at position _X15 .

In some embodiments, the isolated polypeptide comprises a protecting group or spacer, optionally linked to the protecting group at position _X16 .

In some embodiments, an isolated polypeptide comprising a protecting group or spacer optionally linked to a protecting group at position _X1 is conjugated via a lysine at that position.

In some embodiments, an isolated polypeptide comprising a protecting group or spacer optionally linked to a protecting group at position _X15 is conjugated via a lysine at that position.

In some embodiments, an isolated polypeptide comprising a protecting group or spacer optionally linked to a protecting group at position _X16 is conjugated via a lysine at that position.

In some embodiments, the protecting group is selected from the group consisting of acetyl, allyloxycarbonyl, benzyl, Boc, Cbz, Dmb, (dimethyl-2,6-diisoxicyclohex-1-ylidene)ethyl, Fmoc, t-butyl, or trityl. In some embodiments, the protecting group is selected from acetyl, allyloxycarbonyl, dimethyl-2,6-diisoxicyclohex-1-ylidene)ethyl, Fmoc, t-butyl, or trityl. In some embodiments, the protecting group is acetyl, allyloxycarbonyl, dimethyl-2,6-diisoxicyclohex-1-ylidene)ethyl, or Fmoc,

In some embodiments, the spacer is selected from the group consisting of formula XI, formula XII, formula XIII, formula XIV, formula XV, formula XVI, formula XVII, and formula XVIII. In some embodiments, the isolated polypeptide further comprises a spacer of formula XIII. In some embodiments, the isolated polypeptide further comprises a spacer of formula XIV. In some embodiments, the isolated polypeptide further comprises a spacer of formula XVII. In some embodiments, the isolated polypeptide further comprises a spacer of formula XVIII. In some embodiments, the spacer is linked to a protecting group. In some embodiments, the protecting group is selected from the group consisting of acetyl, allyloxycarbonyl, benzyl, Boc, Cbz, Dmb, (dimethyl-2,6-diisoxocyclohex-1-ylidene)ethyl, Fmoc, tert-butyl, or trityl. In some embodiments, the protecting group is acetyl, allyloxycarbonyl, dimethyl-2,6-diisoxocyclohex-1-ylidene)ethyl, Fmoc, tert-butyl, or trityl. In some embodiments, the protecting group is acetyl, trityl, or tert-butyl. In some embodiments, the spacer is not associated with the protecting group.

Illustrative polypeptide intermediates

In some embodiments, an isolated polypeptide of the invention comprises an amino acid sequence selected from the group consisting of the following peptides listed in Table 4:

In some embodiments, the present invention provides a peptide intermediate as shown in Table 4 above. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 129, SEQ ID NO: 144, SEQ ID NO: 145, or SEQ ID NO: 156. In some embodiments, the peptide intermediate has the amino acid sequence of SEQ ID NO: 129. In some embodiments, the peptide intermediate has the amino acid sequence of SEQ ID NO: 145. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 129, SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, or SEQ ID NO: 150. In some embodiments, the peptide intermediate has the amino acid sequence of SEQ ID NO: 147. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, SEQ ID NO: 160, SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, or SEQ ID NO: 170. ID NO: 152, SEQ ID NO: 153, or SEQ ID NO: 154. In some embodiments, the peptide intermediate has the amino acid sequence of SEQ ID NO: 151.

In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 155 or SEQ ID NO: 156. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 155. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 159, or SEQ ID NO: 160. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 157. In some embodiments, the peptide intermediate is a peptide having the amino acid sequence of SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, or SEQ ID NO: 164. In some embodiments, the peptide intermediate is a peptide having amino acid sequence SEQ ID NO: 164.

8. Application, composition and administration

Methods of application

According to another embodiment, the present invention provides a method for treating a metabolic disease or disorder in a subject in need of treatment, comprising providing the subject with an effective amount of an amylin analog polypeptide of the invention or a pharmaceutical composition thereof. Metabolic diseases or disorders include type 1 diabetes, type 2 diabetes, and obesity. The invention further relates to a method for increasing body weight loss in a subject, including a subject with diabetes, comprising providing the subject with an effective amount of an amylin analog polypeptide of the invention.

The amylin analog polypeptides of the present invention are particularly useful for treating diabetes, wherein the method comprises providing a subject with diabetes an effective amount of an amylin analog polypeptide. In some embodiments, the amylin analog polypeptide of the present invention is used to treat a subject with type 1 or type 2 diabetes to control or reduce the subject's blood sugar concentration, wherein the blood sugar level can be controlled or approximated based on measured concentrations of glycated hemoglobin in the blood (hemoglobin A1c, HbA1c).

(i) In some embodiments, the amylin analog polypeptide of the invention is used to treat a subject with type 1 diabetes;

(ii) In some embodiments, the amylin analog polypeptide of the invention is used to treat a subject with type 2 diabetes;

(iii) In some embodiments, the amylin analog polypeptide of the invention is used to treat obesity; and

(iv) In some embodiments, an amylin analog polypeptide of the invention is used to reduce the weight of a subject, such as a subject with diabetes,

wherein the amylin analogue polypeptide of use (i), (ii), (iii) or (iv) comprises any amino acid sequence of the present invention, including one selected from the group consisting of SEQ ID NOS: 199, 200, 204, 206, 127, 57, 128, 129, 43, 209, 130, 64,65, 131, 109 and 27.

In some embodiments, an amylin analog polypeptide is used to treat a subject with type 1 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 27. In some embodiments, an amylin analog polypeptide is used to treat a subject with type 1 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 64. In some embodiments, an amylin analog polypeptide is used to treat a subject with type 1 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 65. In some embodiments, an amylin analog polypeptide is used to treat a subject with type 1 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 131.

In some embodiments, an amylin analog polypeptide is used to treat a subject with type 2 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 27. In some embodiments, an amylin analog polypeptide is used to treat a subject with type 2 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 64. In some embodiments, an amylin analog polypeptide is used to treat a subject with type 2 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 65. In some embodiments, an amylin analog polypeptide is used to treat a subject with type 2 diabetes, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 131.

In some embodiments, an amylin analog polypeptide is used to treat an obese subject, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 27. In some embodiments, an amylin analog polypeptide is used to treat an obese subject, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 64. In some embodiments, an amylin analog polypeptide is used to treat an obese subject, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 65. In some embodiments, an amylin analog polypeptide is used to treat an obese subject, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 131.

In some embodiments, an amylin analog polypeptide is used to reduce the weight of a subject, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 27. In some embodiments, an amylin analog polypeptide is used to reduce the weight of a subject, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 64. In some embodiments, an amylin analog polypeptide is used to reduce the weight of a subject, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 65. In some embodiments, an amylin analog polypeptide is used to reduce the weight of a subject, wherein the amylin analog polypeptide comprises the amino acid sequence of SEQ ID NO 131.

Provided are amylin analog polypeptides of the invention as insulin (i.e., administered) to a subject with diabetes to maintain, control or reduce the concentration of sugar in the blood of the subject. Subjects with diabetes who are treated with an amylin analog polypeptide of the invention, as an adjunct to insulin therapy, are at risk of hypoglycemia (i.e., low blood sugar), especially severe hypoglycemia. Accordingly, reducing the insulin dose during meals in subjects with diabetes when treated with an amylin analog polypeptide of the invention is intended to reduce the risk of hypoglycemia, especially severe hypoglycemia.

Severe hypoglycemia, as used in this document, refers to an episode of hypoglycemia that requires assistance from another person (including assistance with administering oral carbohydrate) or that requires administration of glucagon, intravenous glucose, or other medical intervention.

Accordingly, administration of the amylin analog polypeptide of the invention as an adjunct to insulin therapy, particularly mealtime insulin therapy, typically requires a reduction in the mealtime insulin dose necessary to maintain a healthy subject's blood sugar level at a proper concentration. In other words, diabetics with type 1 or type 2 diabetes who are already self-administering mealtime insulin at a certain dose before initiating treatment with the amylin analog polypeptide of the invention will decrease (e.g., by 25%, 50%, 75%, or 100%) the mealtime insulin dose, they continue to self-administer the amylin analog polypeptide of the invention.

In some embodiments, the method comprises providing an amylin analog polypeptide of the invention or a pharmaceutical composition thereof to a subject in need of treatment by injection. In some embodiments, the method comprises providing an amylin analog polypeptide of the invention or a pharmaceutical composition thereof intended for oral administration to a subject in need of treatment.

In some embodiments, the method comprises providing an amylin analog polypeptide of the invention or a pharmaceutical composition thereof to a subject in need of treatment via implantation. In some embodiments, the method comprises providing continuous delivery from an osmotic delivery device of the amylin analog polypeptide to a subject in need of treatment. The delivery device, such as an osmotic delivery device, contains a sufficient amount of the amylin analog polypeptide of the invention for continuous administration for up to 3 months, 6 months, 9 months, 12 months, 18 months, or 24 months. As such, continuous administration of the amylin analog polypeptide of the invention via an osmotic delivery device eliminates daily or multiple daily dosages of existing amylin analog polypeptides, such as pramlintide. Diabetics who are treated with pramlintide should coordinate the pre-meal dosage of pramlintide with mealtime insulin administered after meals. In contrast, diabetics who receive the amylin analog polypeptide of the invention via osmotic delivery receive continuous delivery of the amylin analog polypeptide and only require reduced doses of meal-induced insulin control.

The substantially stable delivery of the amylin analog polypeptide that is a specific glucagon agonist from the osmotic delivery device is continuous during the period of administration. In some embodiments, the subject or patient is a human subject or human patient.

In some embodiments of the present invention, the period of administration is, for example, at least about 3 months, at least about 3 months to about one year, at least about 4 months to about one year, at least about 5 months to about one year, at least about 6 months to about one year, at least about 8 months to about one year, at least about 9 months to about one year, at least about 10 months to about one year, at least about one year to about two years, at least about two years to about three years.

In other embodiments, the methods of treatment of the present invention provide a significant reduction in the fasting plasma glucose concentration of a subject after implantation of the osmotic delivery device in the subject (relative to the fasting plasma glucose concentration of the subject prior to implantation of the osmotic delivery device), which is achieved within about 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day or less after implantation of the osmotic delivery device in the subject. A significant reduction in fasting plasma glucose is typically statistically significant as demonstrated by an appropriate statistical test or determined to be significant to the subject by a practicing physician. A significant reduction in fasting plasma glucose relative to a pre-implantation baseline is typically maintained during the period of administration.

In some embodiments, the present invention provides a method of treating a disease or condition in a subject in need of treatment. The method includes continuous delivery of a drug from an osmotic delivery device, wherein substantially stable delivery of the drug at therapeutic concentrations is achieved in the subject. The substantially stable delivery of the drug from the osmotic delivery device is continuous over an administration period of at least about 3 months. The drug has a known or determined half-life in a typical subject. Humans are preferred subjects for practicing the present invention. The present invention includes a drug effective for treating a disease or condition, as well as an osmotic delivery device containing the drug for use in the present methods of treating a disease or condition in a subject in need of treatment. Advantages of the present invention include mitigation of drug toxicity associated with peaks and attenuation of suboptimal drug therapy associated with trough concentrations.

In some embodiments, substantial sustained delivery of the drug at therapeutic concentrations is achieved for a period of about 1 month, 7 days, 5 days, 3 days, or 1 day after implantation of the osmotic delivery device into the subject.

The invention also provides a method of promoting weight loss in a subject in need thereof, a method of treating overweight or obesity in a subject in need thereof, and/or a method of suppressing appetite in a subject in need thereof. The method comprises providing delivery of an isolated amylin analog polypeptide. In some embodiments, the isolated amylin analog polypeptide is continuously delivered from an implantable osmotic delivery device. In some embodiments, significant sustained delivery of the isolated amylin analog polypeptide from the osmotic delivery device is achieved and is substantially continuous during the period of administration. In some embodiments, the subject is a human.

The present invention includes an isolated amylin analog polypeptide, as well as an osmotic delivery device comprising the isolated amylin analog polypeptide for use in these methods in a subject in need of treatment. The subject may have type 2 diabetes. The subject may, if desired, have a baseline HbA1c% of greater than 10.0%, i.e., may be a high baseline (HBA) subject. The subject may not have previously received a drug for the treatment of type 2 diabetes.

In other embodiments, the treatment methods of the present invention provide a significant reduction in the fasting plasma glucose concentration of a subject after implantation of the osmotic delivery device in the subject (relative to the fasting plasma glucose concentration of the subject prior to implantation of the osmotic delivery device), which is achieved within about 7 days or less after implantation of the osmotic delivery device in the subject, within about 6 days or less after implantation of the osmotic delivery device in the subject, within about 5 days or less after implantation of the osmotic delivery device in the subject, within about 4 days or less after implantation of the osmotic delivery device in the subject, within about 3 days or less after implantation of the osmotic delivery device in the subject, within about 2 days or less after implantation of the osmotic delivery device in the subject, or within about 1 day or less after implantation of the osmotic delivery device in the subject. In preferred embodiments of the present invention, a significant reduction in the fasting plasma glucose concentration of a subject after implantation of the osmotic delivery device, relative to the fasting plasma glucose concentration of the subject prior to implantation, is achieved within about 2 days or less, preferably within about 1 day or less after implantation of the osmotic delivery device in the subject, or more preferably within about 1 day after implantation of the osmotic delivery device in the subject. A significant reduction in fasting plasma glucose is typically statistically significant as demonstrated by an appropriate statistical test or determined to be significant for the subject by a medical practitioner. A significant reduction in fasting plasma glucose relative to the pre-implantation baseline is typically maintained during the administration period.

In embodiments of all aspects of the present invention relating to methods of treating a disease or condition in a subject, as an example, an osmotic delivery device includes the following: an impermeable reservoir comprising an inner and an outer surface and first and second open ends; a semipermeable membrane sealed at the first open end of the reservoir; an osmotic motor inside the reservoir adjacent to the semipermeable membrane; a piston adjacent to the osmotic motor, wherein the piston forms a movable seal with the inner surface of the reservoir, the piston divides the reservoir into a first chamber and a second chamber, the first chamber contains the osmotic motor; a drug or a suspension formulation containing the drug, wherein the second chamber contains the drug or the suspension formulation, and the drug or the suspension formulation is in liquid form; and a diffusion retarder is inserted into the second open end of the reservoir, the diffusion retarder is adjacent to the suspension formulation. In preferred embodiments, the reservoir comprises titanium or a titanium alloy.

In embodiments of all aspects of the present invention relating to methods of treating a disease or condition in a subject, the medicament may comprise a medicament and a carrier composition. Alternatively, the methods use suspension formulations, they may, for example, include a particulate composition comprising particles of the medicament and a carrier composition. Carrier compositions for use in the suspension formulation of the present invention may, for example, comprise a solvent and a polymer.

The reservoir of the osmotic delivery devices may, for example, comprise titanium or a titanium alloy.

In embodiments of all aspects of the present invention, the implanted osmotic delivery device may be used to provide subcutaneous delivery.

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

Combinations

In some embodiments, an amylin analog polypeptide of the invention is combined with a second agent. In some embodiments, an amylin analog polypeptide of the invention is combined with a second agent, wherein the second agent is an insulinotropic peptide. In some embodiments, an amylin analog polypeptide of the invention is combined with a second agent, wherein the second agent is a GLP-1 receptor agonist. In some embodiments, an amylin analog polypeptide of the invention is combined with a second agent, wherein the second agent is a (GLP-1) agonist, such as exenadide, an exenadide derivative, an exenadide analog, or semaglutide. In some embodiments, the GLP-1 receptor agonist is exenadide. In some embodiments, the GLP-1 receptor agonist is semaglutide.

In some embodiments, an amylin analog polypeptide of the invention without association with a second agent is administered to a subject in combination with a second agent, wherein the second agent is a (GLP-1) agonist such as exenadide, an exenadide derivative, an exenadide analog, or semaglutide.

In some embodiments, an amylin analog polypeptide of the invention is combined with insulin or an insulin derivative. In some embodiments, an amylin analog polypeptide of the invention is combined with long-acting insulin or a long-acting insulin derivative.

In some embodiments, an amylin analog polypeptide of the invention, while not combined with insulin or an insulin derivative, is administered to a subject in combination with insulin or an insulin derivative, i.e., as an adjunct to insulin therapy. In some embodiments, an amylin analog peptide of the invention, while not combined with insulin or an insulin derivative, is administered to a subject in combination with insulin administered with meals. In some embodiments, the subject has type 1 diabetes. In some embodiments, the subject has type 2 diabetes.

In some embodiments, an amylin analog polypeptide of the invention is co-administered to a human patient with insulin or an insulin derivative to provide a so-called dual hormone "artificial pancreas" therapy. In some embodiments, an amylin analog polypeptide of the invention, while not combined with insulin or an insulin derivative, is co-administered to a subject in combination with insulin or an insulin derivative to provide a so-called dual hormone "artificial pancreas" therapy. In some embodiments, an amylin analog polypeptide of the invention is combined with insulin or an insulin derivative and administered to a subject in combination with the insulin or an insulin derivative to provide a so-called dual hormone "artificial pancreas" therapy. In some embodiments, the artificial pancreas therapy comprises a rapid-acting insulin or a rapid-acting insulin derivative. In some embodiments, the artificial pancreas therapy comprises a long-acting or background insulin or a long-acting or background insulin derivative.

In some embodiments, any of the amylin analogs of the invention is combined with exenatide. In some embodiments, any of the amylin analogs of the invention is combined with a GLP-1 receptor agonist.

Some embodiments of the present invention include the use of an amylin analog polypeptide in combination with a second therapeutic agent, such as a second polypeptide such as, by way of non-limiting example, insulinotropic peptides, peptide hormones such as glucagon and incretin mimetics (e.g., a GLP-1 receptor agonist such as exenatide), and peptide analogs and peptide derivatives thereof; PYY (also known as peptide YY, tyrosine-tyrosine peptide), and peptide analogs and peptide derivatives thereof, such as PYY (3-36); oxyntomodulin, and peptide analogs and peptide derivatives thereof); and gastric inhibitory peptide (GIP), and peptide analogs and peptide derivatives thereof. In some embodiments, a pharmaceutical composition comprising an amylin analog polypeptide in combination with a GLP-1 receptor agonist is used to treat type 2 diabetes.

GLP-1, including three peptide forms, GLP-1(1-37), GLP-1(7-37), and GLP-1(7-36)amide, as well as peptide analogs of GLP-1, have been shown to stimulate insulin secretion (i.e., are insulinotropic), which induces glucose uptake into cells and results in a reduction in serum glucose concentrations (see, e.g., Mojsov, S., Int. J. Peptide Protein Research, 40:333-343 (1992)).

Numerous GLP-1 receptor agonists (e.g., peptide derivatives and peptide analogs of GLP-1) that exhibit insulinotropic actions are known in the art (see, 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; 6,593,295; 6,703,359; 6,706,689; 6,720,407; 6,821,949; 6,849,708; 6,849,714; 6,887,470; 6,887,849; 6,903,186; 7,022,674; 7,041,646; 7,084,243; 7,101,843; 7,138,486; 7,141,547; 7,144,863; and 7,199,217), as well as in clinical trials (e.g., taspoglutide and albiglutide). One example of a GLP-1 receptor agonist that may be used in the practice of the present invention is Victoza® (Novo Nordisk A/S, Bagsvaerd D K) (liraglutide; U.S. Patents 6,268,343, 6,458,924, and 7,235,627). The once-daily injectable drug Victoza® (liraglutide) is commercially available in the United States, Europe, and Japan. Another example of a GLP-1 receptor agonist is Ozempic® (Novo Nordisk A/S, Bagsvaerd D K) (semaglutide). For ease of reference, the family of GLP-1 receptor agonists, GLP-1 peptides, peptide derivatives of GLP-1, and peptide analogues of GLP-1 that have insulinotropic activity are collectively referred to as "GLP-1".

The exenatide molecule has the amino acid sequence of exendin-4 (Kolterman O. G., et al., J. Clin. Endocrinol. Metab. 88 (7): 3082-9 (2003)) and is produced by chemical synthesis or recombinant expression. For ease of reference, the exenatide family of peptides (e.g., including exendin-3, exendin-4, and exendin-4-amide), exenatide peptide derivatives, and exenatide peptide analogs are collectively referred to as "exenatide".

Peptide YY (PYY) is a peptide amide consisting of 36 amino acid residues. PYY inhibits motility and blood flow (Laburthe, M., Trends Endocrinol Metab. 1(3): 168–74 (1990), mediates intestinal secretion (Cox, H. M., et al., Br J Pharmacol 101(2):247–52 (1990); Playford, R. J., et al., Lancet 335(8705): 1555–7 (1990)) and stimulates net absorption (MacFayden, R. J., et al., Neuropeptides 7(3):219–27 (1986)). Two major variants have been identified in vivo, PYY(1–36) and PYY(3–36) (e.g., Eberlein, G. A., et al., Peptides 10(4), 797–803 (1989)). The PYY sequence, as well as peptide analogs and peptide derivatives, are known in the art (e.g., U.S. Patent Nos. 5,574,010 and 5,552,520).

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

Gastric inhibitory peptide (GIP) is an insulinotropic peptide hormone (Efendic, S., et al., Horm Metab Res. 36: 742-6 (2004)) and is secreted by the duodenal and jejunal mucosa in response to ingested fat and carbohydrates, which stimulate the pancreas to secrete insulin. GIP circulates as a biologically active peptide of 42 amino acids. GIP is also known as glucose-dependent insulinotropic 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 GIP sequence, as well as its peptide analogs and peptide derivatives, are known in the art (e.g., Meier J.J., Diabetes Metab Res Rev. 21(2):91-117 (2005) and Efendic S., Horm Metab Res. 36(ll-12):742-6 (2004)).

Glucagon is a peptide hormone produced by the alpha cells of the pancreas that increases blood glucose levels. It acts in the opposite way to insulin, which decreases glucose levels. The pancreas produces glucagon when blood glucose levels fall too low. Glucagon stimulates the liver to convert stored glycogen into glucose, which is released into the bloodstream. Blood glucose levels stimulate the release of insulin. Insulin allows glucose to enter and be used by insulin-dependent tissues. Thus, glucagon and insulin are part of a feedback system that keeps blood glucose levels stable.

Pharmaceutically acceptable compositions

According to another embodiment, the invention provides a composition comprising a compound, i.e., an isolated polypeptide, of the invention or a pharmaceutically acceptable derivative thereof, and a pharmaceutically acceptable carrier, adjuvant or vehicle. The amount of the compound in the compositions of the invention is such that it is effective to measurably activate one or more amylin and/or calcitonin receptors in a biological sample or in a patient. In certain embodiments, the amount of the compound in the compositions of the invention is such that it is effective to measurably activate human amylin receptor 3 (hAMY3) and/or human calcitonin (hCTR), in the absence or presence of human serum albumin, in a biological sample or in a patient. In certain embodiments, a composition of the invention is formulated for administration to a patient in need of such a composition. In some embodiments, a composition of the invention is formulated for injection into a patient. In some embodiments, a composition of the invention is formulated for administration to a patient using an implantable delivery device, such as an osmotic delivery device.

The terms "patient" or "subject" as used herein refer to an animal, preferably a mammal, and most preferably a human.

"Pharmaceutically acceptable derivative" means any non-toxic salt, ester, salt of an ester, or other derivative of a compound of this invention that, when administered to a recipient, is capable of providing, directly or indirectly, the compound of this invention or an inhibitory active metabolite or residue thereof.

The isolated polypeptides of the disclosure (also referred to herein as "active compounds"), and derivatives, fragments, analogs, and homologs thereof, 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. As used herein, the term "pharmaceutically acceptable carrier" is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Suitable carriers are described in the latest edition of Remington Pharmaceutical Sciences, a 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, saline, Ringer's solutions, dextrose solution, and 5% human serum albumin. Liposomes and non-aqueous vehicles such as fixed oils may also be used. The use of such vehicles and agents for pharmaceutically active substances is known in the art. Except in cases where any conventional vehicle or agent is incompatible with the active compound, its use in the compositions is contemplated. Additional active compounds may also be included in the compositions.

The pharmaceutical composition of the present invention is formulated to be compatible with the intended route of administration. Examples of routes of administration include parenteral, such as intravenous, intradermal, subdermal, subcutaneous, oral (e.g., inhalation), transdermal (i.e., topical), mucosal, rectal, or combinations thereof. In some embodiments, the pharmaceutical composition or isolated polypeptide of the disclosure is formulated for topical administration. In some embodiments, the pharmaceutical composition or isolated polypeptide of the disclosure is formulated for inhalation administration. In some embodiments, the pharmaceutical composition is formulated for administration via a device or other suitable delivery mechanism suitable for subdermal or subcutaneous implantation and delivering the pharmaceutical composition subcutaneously. In some embodiments, the pharmaceutical composition is formulated for administration via an implantable device that is suitable for subdermal or subcutaneous implantation and delivering the pharmaceutical composition subcutaneously. In some embodiments, the pharmaceutical composition is formulated for administration using an osmotic delivery device, such as an implantable osmotic delivery device that is suitable for subdermal or subcutaneous placement or other implantation and delivers the pharmaceutical composition subcutaneously. Solutions or suspensions used for parenteral administration, intradermal administration, subdermal administration, subcutaneous administration, or a combination thereof may include the following components: a sterile diluent such as water for injection, saline, fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methylparabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates or phosphates, and agents for adjusting tonicity such as sodium chloride or dextrose. The pH value can be adjusted with acids or bases such as hydrochloric acid or sodium hydroxide. The preparation for parenteral use can be enclosed in ampoules, disposable syringes or multiple-dose vials made of glass or plastic.

Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (if water soluble) or dispersions and sterile powders for the preparation of sterile extemporaneous injectable solutions or dispersions. For intravenous administration, suitable carriers include saline, bacteriostatic water, Cremophor EL (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol, etc.) and suitable mixtures thereof. Proper fluidity can be maintained, for example, by using coating materials such as lecithin, by maintaining the required particle size in the case of dispersions and by using surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example parabens, chlorobutanol, phenol, ascorbic acid, thiomersal, etc. In many cases, it is preferable to include isotonic substances in the composition, for example sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride. Prolonged absorption of injection compositions can be ensured by including in the composition a substance that delays absorption, for example aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in a suitable solvent with one or more of the ingredients enumerated above, as required, followed by filtered sterilization. In general, dispersions are prepared by incorporating the active compound in a sterile stock solution which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the methods of preparation include vacuum drying and freeze-drying, which yield a powder of the active ingredient plus any additional active ingredient from a previously sterile-filtered solution thereof.

Compositions for oral administration typically include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purposes of oral therapeutic administration, the active compound can be combined with excipients and used in the form of tablets, troches or capsules. Oral compositions can also be prepared using a carrier liquid for use as a mouthwash, the compound in the liquid carrier being administered orally, swished in the mouth and spat out or swallowed. Pharmaceutically compatible binders and/or auxiliary materials can be included as part of the composition. Tablets, pills, capsules, troches and the like can contain any of the following ingredients or compounds of a similar nature: a binder such as microcrystalline cellulose, tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetener such as sucrose or saccharin; or a flavoring such as peppermint, methyl salicylate, or orange flavor.

For administration by inhalation, the compounds are delivered in the form of an aerosol spray from a pressurized container or dispenser that contains a suitable propellant, e.g. a gas such as carbon dioxide, or from a nebulizer.

Systemic administration can also be carried out through the mucous membranes or through the skin. For transmucosal or transdermal administration, wetting agents suitable for the permeable barrier are used in the formulation. Such wetting agents are generally known in the art and include, for example, substances for administration through the mucosa, detergents, bile salts and derivatives of fusidic acid. Administration through the mucosa can be carried out by using nasal sprays or suppositories. For transdermal administration, the active compounds are formulated as ointments, balms, gels or creams, as is generally known in the art.

In some embodiments, the active components are combined with carriers that will protect the compound from rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters and polylactic acid can be used. Methods for preparing such formulations will be obvious to those skilled in the art. Materials can also be obtained commercially from ALZA Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions can also be used as pharmaceutically acceptable carriers. They 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.

In particular, it may be advantageous to formulate compositions for oral or parenteral administration in dosage unit form for ease of administration and uniformity of dosage. As used herein, dosage unit form refers to physically discrete units suitable as unitary dosages for the subject to be treated, each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect, in association with the required pharmaceutical carrier. The specifications for dosage unit forms according to the invention are dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect achieved, and the limitations inherent in the art of formulating compounds, for example, an active compound for the treatment of individuals.

Pharmaceutical compositions may be included in a container, package or dispenser together with instructions for use.

Compositions from medicinal herbs

In some embodiments, there is provided a pharmaceutical composition comprising any of the described polypeptides formulated as a trifluoroacetic acid salt, an acetic acid salt, or a hydrochloride salt. In some embodiments, there is provided a pharmaceutical composition comprising any of the described polypeptides formulated as a trifluoroacetic acid salt. In some embodiments, there is provided a pharmaceutical composition comprising any of the described polypeptides formulated as an acetic acid salt. In some embodiments, there is provided a pharmaceutical composition comprising any of the described polypeptides formulated as a hydrochloride salt.

The compounds, i.e., the isolated polypeptides or pharmaceutically acceptable salts thereof for use in the practice of the present invention, are typically added to particle formulations that are used to make particles containing the polypeptide, which are uniformly suspended, dissolved or dispersed in a suspension carrier to form a suspension formulation. In some embodiments, the amylin analog polypeptide is formulated into a particle formulation and converted (e.g., spray dried) into particles. In some embodiments, the particles containing the amylin analog polypeptide are suspended in the carrier formulation, resulting in the formation of a suspension formulation and suspended particles containing the amylin analog polypeptide.

Preferably, the particulate compositions can be formed into particles using methods such as spray drying, lyophilization, desiccation, freeze drying, milling, granulation, ultrasonic atomization, crystallization, precipitation, or other methods available in the art for forming particles from a mixture of components. In one embodiment of the invention, the particles are spray dried. The particles preferably have substantially the same shape and size.

In some embodiments, the present invention provides drug particle formulations for pharmaceutical use. The particle formulation typically contains a drug and includes 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 amounts of stabilizers in the particle formulation can be determined experimentally based on the activity of the stabilizers and the desired characteristics of the composition in accordance with the teaching of the present disclosure.

In any of the embodiments of the invention, the particulate composition may comprise from about 50% by weight to about 90% by weight of the drug, from about 50% by weight to about 85% by weight of the drug, from about 55% by weight to about 90% by weight of the drug, from about 60% by weight to about 90% by weight of the drug, from about 65% by weight to about 85% by weight of the drug, from about 65% by weight to about 90% by weight of the drug, from about 70% by weight to about 90% by weight of the drug, from about 70% by weight to about 85% by weight of the drug, from about 70% by weight to about 80% by weight of the drug, or from about 70% by weight to about 75% by weight of the drug.

Generally, the amount of carbohydrates in the particle composition is determined by aggregation problems. In general, the amount of carbohydrates should not be too high to avoid stimulating crystal growth in the presence of water due to excess carbohydrates not associated with the drug.

Typically, the amount of antioxidant in a particle formulation is determined by oxidation issues, while the amount of amino acids in the formulation is determined by oxidation issues and/or particle formability during spray drying.

Typically, the amount of buffer in a particle formulation is determined by pre-treatment issues, stability issues, and particle deformability during spray drying. Buffer may be required to stabilize the drug during processing, such as solution preparation and spray drying, when all stabilizers are solubilized.

Examples of carbohydrates that may be included in the particulate composition 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, melezitose, maltodextrins, dextrans, and starches), and polyols (acyclic polyols such as mannitol, xylitol, maltitol, lactitol, xylitol-sorbitol, pyranosyl sorbitol, and myoinositol). Suitable carbohydrates include disaccharides and/or non-reducing sugars such as sucrose, trehalose, and raffinose.

Examples of antioxidants that may be included in the particulate composition 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. Additionally, amino acids that are easily oxidized may be used as antioxidants, such as cysteine, methionine, and tryptophan.

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

Examples of buffers that may be included in the particle formulation include, but are not limited to, citrate, histidine, succinate, phosphate, maleate, TRIS, acetate, carbohydrate buffer, and gly-gly. Suitable buffers include citrate, histidine, succinate, and TRIS.

Examples of inorganic compounds that may be included in the particulate composition _include , but are not limited to _, NaCl, _Na2SO4 , _NaHCO3 , KCl, _KH2PO4 , _CaCl2 , and _MgCl2 .

Additionally, the particle composition may include other stabilizers/auxiliaries such as surfactants and salts. Examples of surfactants include, but are not limited to, polysorbate 20, polysorbate 80, PLURONIC® (BASF Corporation, Mount Olive, N.J.) F68, and sodium dodecyl sulfate (SDS). Examples of salts include, but are not limited to, sodium chloride, calcium chloride, and magnesium chloride.

The particles are typically of a size that can be delivered by an implantable osmotic delivery device. Similar particle shape and size typically help to provide a consistent and uniform release rate from such a delivery device; however, a particulate preparation may also be used, the particles of which have an abnormal particle size distribution profile. For example, in a typical implantable osmotic delivery device having a delivery opening, the particle size is less than about 30%, more preferably less than about 20%, more preferably less than about 10% of the diameter of the delivery opening. In one embodiment, a particulate composition for use with an osmotic delivery system, in which the diameter of the delivery opening of the implant is about 0.5 mm, the particle size may be, for example, from less than about 150 microns to about 50 microns. In one embodiment, a particulate composition for use with an osmotic delivery system, in which the diameter of the implant delivery opening is about 0.1 mm, the particle size can be, for example, from less than about 30 microns to about 10 microns. In one embodiment, the opening size is about 0.25 mm (250 microns), and the particle size is from about 2 microns to about 5 microns.

It will be understood by those skilled in the art that a population of particles conforms to principles of particle size distribution. Methods widely used in the art to describe particle size distributions include, for example, average diameters and D values, such as D50 values, which are commonly used to represent the average diameter over a range of particle sizes of a given sample.

The particles of the particulate composition have a diameter of from about 2 microns to about 150 microns, such as less than 150 microns in diameter, less than 100 microns in diameter, less than 50 microns in diameter, less than 30 microns in diameter, less than 10 microns in diameter, less than 5 microns in diameter, and about 2 microns in diameter. Preferably, the particles have a diameter of from about 2 microns to about 50 microns.

Particles of the composition of particles comprising an isolated amylin analog polypeptide, which is an analog, have an average diameter in the range from about 0.3 microns to about 150 microns. Particles of the composition of particles comprising an isolated polypeptide, which is an amylin analog polypeptide, have an average diameter in the range from about 2 microns to about 150 microns, such as 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 about 2 microns. In some embodiments, the particle has an average diameter of from about 0.3 microns to 50 microns, such as between about 2 microns to about 50 microns. In some embodiments, the particles have an average diameter of from 0.3 microns to 50 microns, such as between about 2 microns to about 50 microns, wherein each particle has a diameter of less than about 50 microns.

Typically, the particles of the particulate compositions, when introduced into a suspension vehicle, do not settle for less than about 3 months, preferably do not settle for less than about 6 months, more preferably do not settle for less than about 12 months, more preferably do not settle for less than 24 months at the delivery temperature, and most preferably do not settle for less than 36 months at the delivery temperature. Suspension vehicles typically have a viscosity of about 5,000 to about 30,000 poise, preferably about 8,000 to about 25,000 poise, more preferably about 10,000 to 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 typically have a lower level of settling in viscous suspension vehicles than larger particles. Accordingly, particles with sizes in the order of microns to nanoscale are generally desirable. In a viscous suspension carrier, particles of about 2 microns to about 7 microns according to the present invention will not settle for at least 20 years at room temperature based on simulation studies. In one embodiment of a particulate composition of the present invention for use in an implantable osmotic delivery device, the composition comprises particles of less than about 50 microns, more preferably less than about 10 microns, more preferably in the range of about 2 microns to about 7 microns.

Thus, the disclosed polypeptides or their pharmaceutically acceptable salts are formulated into dry powders in solid particles that retain maximum chemical and biological stability of the drug. The particles provide long-term stability when stored at high temperature, and therefore allow for the delivery of a stable and biologically effective drug to a subject over a long period of time. The particles are suspended in suspension carriers for administration to patients.

Particles suspended in carriers

In one aspect, a suspension carrier provides a stable environment in which a particulate drug formulation is dispersed. The particulate drug formulations are chemically and physically stable (as described above) in the suspension carrier. The suspension carrier typically includes 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 carrier may include additional components, including, but not limited to, surfactants, antioxidants, and/or other compounds soluble in the carrier.

The viscosity of the suspension carrier is generally sufficient to prevent sedimentation of the drug particle formulation during storage and for use in a delivery method, such as an implantable, osmotic delivery device. The suspension carrier is biodegradable because the suspension carrier disintegrates or breaks down 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 particle is absorbed.

In embodiments, the suspension carrier is a "single phase" suspension carrier, which is a solid, semi-solid, or liquid homogeneous system that is physically and chemically uniform throughout its volume.

The solvent in which the polymer is dissolved can affect the characteristics of the suspension composition, 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 carrier exhibits phase separation upon contact with an aqueous medium. In some embodiments of the present invention, the solvent can be selected in combination with the polymer such that the resulting suspension carrier exhibits phase separation upon contact with an aqueous medium having less than about 10% water.

The solvent may be a suitable solvent that is immiscible with water. The solvent may also be selected such that the polymer is soluble in the solvent at high concentrations, such as polymer concentrations greater than about 30%. Examples of solvents useful in the practice of 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," i.e., have a low moisture content. Preferred solvents for use in making the suspension vehicle include lauryl lactate, lauryl alcohol, benzyl benzoate, and mixtures thereof.

Examples of polymers for making the suspension carrier of the present invention include, but are not limited to, a polyester (e.g., polylactic acid and a copolymer of polylactic acid and polyglycolic acid), a polymer containing pyrrolidones (e.g., polyvinylpyrrolidone having a molecular weight in the range of about 2,000 to about 1,000,000), an ester or ether of an unsaturated alcohol (e.g., vinyl acetate), a block copolymer of polyoxyethylene and polyoxypropylene block copolymer, or a mixture 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 having 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 7,900-10,800). The polymer used in the suspension vehicle may include one or more different polymers or may include different types of a single polymer. The polymer used in the suspension vehicle may also be dry or have a low moisture content.

Generally speaking, the suspension carrier for use in the present invention may vary in composition based on the desired performance characteristics. In one embodiment, the suspension carrier may comprise from about 40 wt. % to about 80 wt. % polymer(s) and from about 20 wt. % to 60 wt. % polymer(s). Preferred embodiments of the suspension carrier include carriers made from a combination of polymer(s) and solvent(s) in the following ratios: about 25 wt. % solvent and about 75 wt. % polymer; about 50 wt. % solvent and about 50 wt. % polymer; about 75 wt. % solvent and about 25 wt. % polymer. Accordingly, in some embodiments, the suspension carrier may comprise selected components, and in other embodiments, consists essentially of the selected components.

Suspension carriers may exhibit Newtonian viscosity. Suspension carriers are typically formulated to provide a viscosity that maintains a uniform dispersion of the particles of the formulation for a predetermined period of time. This helps to facilitate the preparation of a suspension formulation with the ability to provide controlled delivery of a drug contained in the drug particle formulation. The viscosity of a suspension carrier may vary depending on the desired application, the size and type of the particle formulation, and the loading of the particle formulation in the suspension carrier. The viscosity of a suspension carrier may be altered by changing the type or relative amount of the solvent or polymer used.

The suspension carrier may have a viscosity in the range of about 100 poise to 1,000,000 poise, preferably about 1,000 poise to 100,000 poise. In preferred embodiments of the invention, the suspension carriers typically have a viscosity at 33°C of about 5,000 to about 30,000 poise, preferably about 8,000 to about 25,000 poise, more preferably about 10,000 to about 20,000 poise. In one embodiment, the suspension carrier has a viscosity of about 15,000 poise, plus or minus about 3,000 poise, at a temperature of 33°C. The viscosity can be measured at 33°C, at a shear rate of 10-4/s, using a parallel-plate gap rheometer.

The suspension carrier may exhibit phase separation upon contact with an aqueous medium; however, the suspension carrier typically does not exhibit substantially any phase separation as a function of temperature. Thus, for example, at a temperature in the range of about 0°C to about 70°C and during temperature cycling, for example, cycling the temperature from 4°C to 37°C to 4°C, the suspension carrier typically does not exhibit phase separation.

The suspension carrier can be prepared by combining the polymer and solvent under dry conditions, such as in a dry box. The polymer and solvent can be combined at an elevated temperature, such as about 40°C to about 70°C, and allowed to liquefy and form a single phase. The ingredients can be mixed under vacuum to remove air bubbles released from the dry ingredients. The ingredients can be combined using a conventional mixer, such as a double helical paddle mixer or similar, with a speed set at about 40 rpm. However, higher speeds can also be used to mix the ingredients. After a liquid solution is formed from the ingredients, the suspension carrier can be cooled to room temperature. Differential scanning calorimetry (DSC) data can be used to confirm that the suspension carrier is a single phase. Additionally, the components of the carrier (e.g., the solvent and/or polymer) can be treated to substantially reduce or substantially remove peroxides (e.g., by treatment with methionine, see, e.g., U.S. Patent Application Publication No. 2007-0027105).

The drug particle composition is added to a suspension carrier to form a suspension composition. In some embodiments, the suspension composition may comprise the drug particle composition and a suspension carrier, and in other embodiments, it consists essentially of the drug particle composition and a suspension carrier.

The suspension formulation can be prepared by dispersing the particulate formulation in a suspension carrier. The suspension carrier can be heated and the particulate formulation added to the suspension carrier under dry conditions. The ingredients can be mixed under vacuum at an elevated temperature, such as from about 40°C to about 70°C. The ingredients can be mixed at a sufficient speed, such as from about 40 rpm to about 120 rpm, and for a sufficient amount of time, such as about 15 minutes, to achieve a uniform dispersion of the particulate formulation in the suspension carrier. The mixer can be a double helical paddle mixer or other 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 allowed to cool to room temperature until further use, such as insertion into an implantable drug delivery device, a single dose container, or a multi-dose container.

The suspension composition typically has a total moisture content of less than about 10% by weight, preferably less than about 5% by weight, and more preferably less than about 4% by weight.

In preferred embodiments, the suspension formulation of the present invention is substantially uniform and fluid to allow delivery of the drug particulate formulation from the osmotic delivery device to the subject.

Thus, the components of the suspension carrier provide biocompatibility. The components of the suspension carrier offer appropriate physicochemical properties for the formation of stable suspensions of drug particle formulations. Such properties include, but are not limited to, the following: suspension viscosity; carrier purity; residual carrier moisture; carrier density; dry powder compatibility; implantable device compatibility; polymer molecular weight; carrier stability; and carrier hydrophobicity and hydrophilicity. Such properties can be manipulated and controlled, for example, by changing the composition of the carrier and manipulating the ratio of components used in the suspension carrier.

The suspension formulations described herein can be used in an implantable osmotic delivery device to provide continuous, controlled, and sustained delivery of a zero-order compound over an extended period of time, such as several weeks, months, or up to about one year or more. Such an implantable osmotic delivery device is typically capable of delivering a suspension formulation containing a drug at a desired flow rate over a desired period of time. The suspension formulation can be placed in the implantable osmotic delivery device using conventional methods.

Implantable delivery

The dose and delivery rate can be selected to achieve the desired drug concentration in the blood, typically within less than about 6 half-lives of the drug in the subject's body after implantation of the device. The drug concentration in the blood is selected to provide the optimal therapeutic effect of the drug, while avoiding unwanted side effects that may be caused by excess drug concentration, while at the same time avoiding peak and trough concentrations that may cause side effects associated with peak and trough drug concentrations in plasma.

The implantable osmotic delivery device typically includes a reservoir having at least one opening through which the suspension formulation is delivered. The suspension formulation may be stored in the reservoir. In a preferred embodiment, the implantable drug delivery device is an osmotic delivery device in which drug delivery is accomplished by osmosis. Certain osmotic delivery devices and components thereof have been described, such as the DUROS® delivery device or similar devices (see, e.g., U.S. Patents 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; 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 Publication Nos. 2005/0175701, 2007/0281024, 2008/0091176 and 2009/0202608).

An osmotic delivery device typically consists of a cylindrical reservoir that contains an osmotic motor, a piston, and a drug-containing formulation. The reservoir is closed at one end by a rate-controlled semipermeable membrane and at the other end by a diffusion retarder through which the drug-containing formulation in suspension is released from the drug reservoir. The piston separates the drug formulation from the osmotic motor and uses a seal to prevent water from entering the osmotic motor compartment into the drug reservoir. The diffusion retarder is designed, in combination with the drug-containing formulation, to prevent body fluids from entering the drug reservoir through the opening.

The osmotic device releases the drug at a predetermined rate based on the principle of osmosis. The extracellular fluid enters the osmotic device for delivery through a permeable membrane of the floor directly into the salt engine, which expands to drive the piston at a slow and even delivery rate. The movement of the piston pushes the drug through the opening or outlet at a predetermined shear rate. In one embodiment of the present invention, the reservoir of the osmotic device is filled with a composition in the form of a suspension, and the device is capable of delivering the suspension composition to the subject over an extended period of time (e.g., about 1, about 3, about 6, about 9, about 10 and about 12 months) at a predetermined, therapeutically effective delivery rate.

The rate of release of the drug from the osmotic delivery device typically delivers a predetermined target dose of the drug to the subject, such as a therapeutically effective daily dose delivered over a 24-hour period; that is, the rate of release of the drug from the device provides a substantially steady delivery of the drug to the subject at a therapeutic concentration.

Typically, for an osmotic delivery device, the volume of the useful agent chamber containing the composition with the useful agent is from about 100 μl to about 1000 μl, more preferably from about 120 μl to 500 μl, more preferably between about 150 μl and about 200 μl.

Typically, the osmotic delivery device is implanted in a subject, such as subdermally or subcutaneously, to provide subcutaneous delivery of a drug. The device(s) can be implanted subdermally or subcutaneously in one or both arms (e.g., in the inner part, outer part, or back of the upper arm) or in the abdominal cavity. Preferred sites in the abdominal region are under the skin of the abdomen in the area below the ribs and above the belt line. To provide a number of sites for implanting one or more osmotic delivery devices within the abdominal cavity, the abdominal wall can be divided into 4 quadrants as follows: the upper right quadrant is located at least 2-3 centimeters below the right rib, such as at least about 5-8 centimeters below the right rib, and at least 2-3 centimeters to the right of the abdominal midline, such as at least about 5-8 centimeters to the right of the abdominal midline; the lower right quadrant is located at least 2-3 centimeters above the belt line, for example, at least about 5-8 centimeters above the belt line, and at least 2-3 centimeters to the right of the abdominal midline, for example, at least about 5-8 centimeters to the right of the abdominal midline; the upper left quadrant is located at least 2-3 centimeters below the left rib, for example, at least about 5-8 centimeters below the left rib, and at least 2-3 centimeters to the left of the abdominal midline, for example, at least about 5-8 centimeters to the left of the abdominal midline; and the lower left quadrant is located at least 2-3 centimeters above the belt line, for example, at least about 5-8 centimeters above the belt line, and at least 2-3 centimeters to the left of the abdominal midline, for example, at least about 5-8 centimeters to the left of the abdominal midline. This division provides multiple available sites for implantation of one or more devices at one or more times. Implantation and removal of osmotic delivery devices are typically performed by health care professionals using local anesthesia (eg, lidocaine).

Discontinuation of treatment by removal of the osmotic delivery device from the subject is simple and provides the important advantage of immediate cessation of drug delivery to the subject.

Preferably, the osmotic delivery device has a fail-safe mechanism to prevent unintentional over-delivery or bolus delivery of the drug in such a theoretical situation as clogging or plugging of the outlet part (diffusion retarder) through which the drug is delivered. To prevent unintentional over-delivery or bolus delivery of the drug, the osmotic delivery device is designed and manufactured in such a way that the pressure required to partially or completely displace or separate the diffusion retarder from the reservoir exceeds the pressure required to partially or completely displace or separate the semipermeable membrane in the amount necessary to relieve the pressure in the reservoir. In such a case, the pressure will build up inside the device until it pushes the semipermeable membrane out at the other end, thereby relieving the osmotic pressure. The osmotic delivery device is then static and will no longer deliver the drug, provided that the piston is tightly sealed against the reservoir.

Suspension formulations may also be used in infusion pumps, such as the ALZET® osmotic pumps (DURECT Corporation, Cupertino, CA), which are miniature infusion pumps for continuous dose administration to laboratory animals (e.g., mice and rats).

Examples

The following examples are provided to provide those skilled in the art with a complete disclosure and description of how to practice the present invention, but they are not intended to limit the scope of what applicants consider to be their invention. Care has been taken to maintain accuracy with respect to the figures used (e.g., amounts, concentrations, and percentage changes), but some experimental errors and variations should be taken into account. Unless otherwise noted, temperatures are in degrees Celsius and pressures are at or near atmospheric pressure.

Example 1: Preparation of amylin analog polypeptides

The amylin analog polypeptides of the invention provided in Table 3 were synthesized on a Prelude peptide synthesizer (Protein Technologies Inc., Tucson, AZ) by solid phase methods using the FMOC strategy with activation of N-[(dimethylamino)-1H-1,2,3-triazole-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminhexafluorophosphate N-oxide (HATU) or 2-(6-chloro-1-H-benzotriazol-1-yl)-1,1,3,3-tetramethylaminohexafluorophosphate (HCTU) (5-fold molar excess over amino acids) in N,N-dimethylformamide (DMF), and N'N-diisopropylethylamine (DIEA) was used as a backbone. A 20% piperidine/DMF solution was used to remove the Fmoc protecting group. Rink Amide MBHA LL (Novabiochem) was used as a resin with a loading of (0.30-0.40) mmol/g on a scale of (20-400) μmol.

Final deprotection and cleavage of the peptide from the solid support was accomplished by treating the resin with 92.5% TFA, 2.5% phenol, 2.5% water, and 2.5% triisopropylsilane for 2-3 h. The cleaved peptide was precipitated with cold diethyl ether. The diethyl ether was decanted and the solids were triturated again with cold diethyl ether and pelleted by centrifugation. The crude solids were then dissolved in 1:1 ACN/water, 0.01% TFA. Disulfide bridge formation was accomplished by adding iodine/acetic acid solution (35 mg/mL) to the crude product solution until the solution remained amber in color. The reaction solution was stirred until LC/MS analysis indicated the reaction was complete. A 2% solution of ascorbic acid in _H2O was added until the solution became clear. The final solution of crude product was lyophilized in preparation for final purification.

The lyophilized solid was re-dissolved in 1:1 acetonitrile/water with 0.1% TFA (10-15 mL), purified by reversed-phase HPLC on a Waters XBridge™ BEH 130, CIS, 10 pm, 130 30 X 250 mm ID, using a 30 gradient with the ranges acetonitrile/water 5-75% with 0.1% TFA over 30-45 minutes at a flow rate of 30 ml/min, λ - 215 nm.

Example 2: Purification and characterization of amylin analog polypeptides, i.e., a linear polypeptide, without any lipophilic substituent and an optional spacer

The purified product was lyophilized and analyzed by ESI-LC/MS and analytical HPLC and was shown to be pure (>98%). All masses obtained were consistent with the calculated values.

Characterization of the peptide analogs was performed by C18 HPLC, LC/MS analysis (Acquity SQD Waters Corp., Milford, MA), and UV detection performed by dual absorption signals at 215 nm and 280 nm using one of three methods: method A, method B, method C, or method D.

Method A, LC/MS conditions: performed using a Phenomenex UPLC Aeris™ Peptide XB CI8 35, 1.7 pm, 2.1 X 100 mm or an ACQUiTY VEN300 or VEN130 CT8, 1.77 pm, 2.1 X 100 mm column using 5-65% acetonitrile/water with 0.05% TFA for 30 min at a flow rate of 0.5 mL/min, λ - 215 nm, 280 nm.

Method B, C18 HPLC conditions: UPLC analysis was performed on an Acquity BEH130, C18, 1.7 µm, 100 x 2.10 mm column at 25°C, 5-65% acetonitrile/water with 0.05% TFA for 30 min, flow rate 0.5 mL/min, 1215 nm, 1280 nm.

Method C, UPLC conditions: UPLC analysis was performed on an Acquity BEH130, C18, 1.7 µm, 100 x 2.10 mm column at 25°C, 5-65% acetonitrile/water with 0.05% TFA for 20 min, flow rate 0.5 mL/min, 1215 nm, 1280 nm.

Method D, UPLC conditions: UPLC analysis was performed on an Acquity BEH130, C18, 1.7 μm, 100 X 2.10 mm column at 25°C, 5-65% acetonitrile/water with 0.05% TFA for 10 min, flow rate 0.5 mL/min, 1215 nm, 1280 nm 5.0 μL of sample was injected using PLNO injection mode (partial cycle w/overfilled needle).

Table 3 provides exemplary amylin analog polypeptides of the invention.

Analog polypeptides lacking a lipophilic substituent and an optional spacer are sometimes referred to herein as "linear polypeptides." Analog polypeptides having at least one covalently linked lipophilic substituent and an optional spacer are sometimes referred to herein as "conjugated polypeptides."

Example 3: Synthesis of amylin analog polypeptide intermediates

Synthesis of polypeptides with modifications at positions D-Lysl6 or L-Lysl6, for example (compounds B2, A129, B4 and B8)

Upon completion of the synthesis of the linear polypeptide described in Example 1, the resin was washed with dichloromethane (DCM) and dried under vacuum for 30 min. For analogs containing the Alloc protecting group, removal was facilitated with a solution of Pcl(PPn3)3 in (chloroform/acetic acid/n-methylmorpholine, 37:2:1). The resulting deprotected resin was washed with 2% sodium diethyl dithiodicarbamate trihydrate/DMF (6 x 30 sec), 2% DIEA/DMF (6 x 30 sec), and finally DMF (6 x 30 sec). Extension of the spacer region was performed in a stepwise manner with manual addition of each building block under preactivation conditions. To 1 mL of 200 mmol Fmoc-γGlu-(OH)-OtBu in DMF was added 0.5 mL DIEA (800 mmol) followed by 0.5 mL HCTU (400 mmol) and the resulting reaction solution was stirred for 5 min where it was added to the linear residue on the linear sequence. The reaction mixture was stirred under nitrogen for 30 min. The resin was dried and washed with DMF (6 x 30 sec). Subsequent deprotection of the Fmoc protecting group was facilitated using 20% piperidine/DMF followed by a final wash with DMF (6 x 30 sec). Final deprotection and cleavage of the peptide from the solid support was accomplished by treating the resin (95% TFA, 2% water, 2% thioanisole, 1% triisopropylsilane) for 2-3 h. As each building block was incorporated using the method described above, each intermediate was isolated and characterized by HPLC/MS.

Synthesis of polypeptides with modifications at the N-terminus, for example, (A130, B22 and B23)

Synthesis of the linear sequence was performed as described in Example 1. Addition of the albumin binding moiety was facilitated by removal of the N-terminal Fmoc protecting group with 20% piperidine/DMF. The resin was washed with DMF and incorporation of the side chain building blocks was performed stepwise under preactivation conditions. To 1 mL of 200 mmol Fmoc-γGlu-(OH)-OtBu in DMF was added 0.5 mL of DIEA (800 mmol) followed by 0.5 mL of HCTU (400 mmol). The resulting reaction solution was stirred for 5 min while adding it to the linear sequence. The reaction mixture was stirred under nitrogen for 30 min. The resin was dried and washed with DMF (6 x 30 sec). Final deprotection and cleavage of the peptide from the solid support was accomplished by treating the resin with (95% TFA, 2% water, 2% thioanisole, 1% triisopropylsilane) for 2-3 h. Each intermediate was isolated and characterized by HPLC/MS. Chemical data for the internally modified polypeptides and N-terminal modified polypeptide intermediates are listed in Table 5 below.

Example 4. Covalent attachment of a lipophilic substituent and an optional spacer to amylin analog polypeptides, i.e., conversion of linear polypeptides into conjugated polypeptides

Synthesis of amylin analog polypeptides conjugated to one or more lipophilic albumin-binding substituents and an optional spacer was carried out with modifications to the synthetic method described in Example 1.

Upon completion of the synthesis of the linear polypeptide described in Example 1, the resin was washed with dichloromethane (DCM) and dried under vacuum for 30 min. For analogs containing the Alloc protecting group, removal was facilitated with a solution of Pd( _PPh3 ) ₃ in (chloroform/acetic acid/n-methylmorpholine, 37:2:1). For analogs containing BOC-Lys(Fmoc)-OH, the Fmoc protecting group was removed using 20% piperidine/DMF. The resulting unprotected resin was washed with DMF (6 x 30 sec). Extension of the spacer region was then performed in a stepwise manner with manual addition of each building block under preactivation conditions. Addition of the lipophilic substituent (also called the “acyl chain”) was performed under normal SPPS conditions without a preactivation step. Final deprotection and cleavage of the peptide from the solid support was accomplished by treating the resin with (95% TFA, 2% water, 2% thioanisole, 1% triisopropylsilane) for 2-3 h. The cleaved peptide was precipitated with cold diethyl ether. The diethyl ether was decanted, and the solids were triturated again with cold diethyl ether and pelleted by centrifugation.

The crude product was then dissolved in ACN/ _H2O , 0.1% TFA. Iodine/acetic acid solution (35 mg/mL) was added to each crude peptide solution until the solution was consistently amber in color. The reaction solution was stirred until LC/MS analysis indicated that the desired disulfide bridge had formed. A 2% ascorbic acid solution in _H2O was added to the reaction solution until the solution became clear. The solution was frozen and lyophilized. Purification was accomplished using the methods described in Example 1.

An illustrative synthesis of the conjugated peptide is described. Synthesis of A109: The linear sequence was synthesized as described in Example 1. Addition of the albumin binding moiety was facilitated by removal of the N-terminal Fmoc protecting group with 20% piperidine/DMF. The resin was then washed with DMF and incorporation of side chain building blocks was performed stepwise under preactivation conditions. To 1 mL of 200 mmol Fmoc-γGlu-(OH)-OtBu in DMF was added 0.5 mL of DIEA (800 mmol) followed by 0.5 mL of HCTU (400 mmol). The resulting reaction solution was stirred for 5 min while adding it to the linear sequence. The reaction mixture was stirred under nitrogen for 30 min. The resin was then dried and washed with DMF (6 x 30 sec). The Fmoc γGlu protecting group was removed with 20% piperidine/DMF followed by coupling with octadecanedioic acid (C18) (200 mmol) in DMF using HATU (400 mmol) and DIEA (800 mmol) under normal solid phase conditions.

Cleavage was accomplished using 95% TFA/2% water/2% thioanisole/1% TIPS. The crude product was then dissolved in ACN/ _H2O , 0.1% TFA. Iodine/acetic acid solution (35 mg/mL) was added to the crude peptide solution until the solution remained amber in color. The reaction solution was stirred until LC/MS analysis indicated that the desired disulfide bridge had formed. 2% ascorbic acid in _H2O was added to the reaction solution until the solution became clear.

Lyophilization of the crude product yielded an off-white solid, which was purified by the methods described in Example 2.

Example 5: Stability of amylin analog polypeptides

Several of the amylin analog polypeptides described herein were tested as the acetate salt of trifluoroacetate for stability in DMSO (i.e., an organosulfur solvent) or aqueous (i.e., DI water) at 1 mg/mL. These analog polypeptides were incubated at 37°C and samples were removed at various time intervals and analyzed by LC/MS and HPLC to determine the purity and mass of the parent peptide and the extent of any degradation products. The purity results from these analyses are provided in Tables 6A and 6B and are considered indicative of stability.

Example 6: Solubility of amylin analog polypeptides

The analogue polypeptides described in this document were tested for solubility in 20% DMSO saline in water (i.e., bioassay buffer) or water (DI water) at room temperature. Samples were visually inspected for sample clarity and any appearance of turbidity or cloudiness. The results of this analysis are presented in Table 7.

Example 7: Functional assays: human calcitonin receptor and amylin receptor 3

Activation of the human calcitonin receptor (hCTR) or the human amylin 3 receptor (hAMY3R) results in an increase in cellular cyclic adenosine monophosphate (cAMP). In the presence of the non-specific phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) cAMP/cGMP, cAMP accumulation can be measured in vitro using common detection methods. Thus, the in vitro potency (pEC50) can be estimated for peptides activated by each of these receptors using dose-response curves for cAMP accumulation.

Cells, culture and cAMP analysis

HEK293-CNG cells stably expressing human calcitonin receptor (hCTR) or co-expressing human calcitonin receptor and human activity modifying protein 3 receptor (hAMY3R) (Codex Biosolutions #CB-80200-258 and #CB-80-200-271, respectively) were grown in 90% DMEM, 10% FBS, 250 μg/ml G418, and 1 μg/ml puromycin (hCTR cells) or 90% DMEM, 10% FBS, 250 μg/ml G418, 1 μg/ml puromycin, and 150 μg/ml hygromycin B (hAMY3R cells). Cells were cultured in growth media for no more than 10 passages prior to testing.

On the day of the assay, hCTR- or hAMY3R-expressing cells were counted and split at 500 cells per well into white 384-well OptiPlates (PerkinElmer #6007299) in 5 µl of stimulation buffer consisting of 1X HBSS, 5 mM HEPES, 0.5 mM IBMX, and 0.1% bovine serum albumin (BSA) or 1X HBSS, 5 mM HEPES, 0.5 mM IBMX, and 0.1% casein with 0%, 0.1%, or 4% human serum albumin (HSA).

Peptides were serially diluted in the same buffer as above for each given assay condition. Two assay control solutions consisting of 50 μM forskolin (maximum cAMP) or assay buffer alone (minimum cAMP) were also prepared in the respective stimulation buffers. Five microliters of each peptide or assay control concentration were added in triplicate to the wells and incubated for thirty minutes at room temperature. During this step, incubations with 4x labeled cAMP and 4x Ulight® anti-cAMP (consisting of a monoclonal antibody to cAMP fused to Ulight™ dye) were performed according to the manufacturer's protocol (PerkinElmer LANCE Ultra cAMP kit). Following this incubation, 5 μl europium-labeled cAMP and 5 μl Ulight anti-cAMP were added to the wells. The plates were covered with an adhesive cap to prevent evaporation and incubated for 60 minutes at room temperature in the dark. The plates were read on an Envision fluorescent plate reader (PerkinElmer).

Data Analysis

Test values were first normalized to the mean systemic forskolin-induced maximum and minimum cAMP values using Excel using the formula: (test value - systemic min.avg) / (systemic max.avg - systemic min.avg) * 100. Normalized test values represent the original adjusted percentage of the systemic forskolin-induced maximum cAMP response. Normalized data from triplicate tests were analyzed and used to estimate the EC50 for each peptide at each receptor. Data were fitted in GraphPad Prism (v7.04) using a 4-parameter logistic curve model Y = bottom + (top - bottom) / (l + 10 ^∧ ((LogEC50 - X))). The Hill slope was clamped to 1.0. EC50 values were converted to pEC50 values using the formula: pEC50=-Log(EC50).

Interpretation of data

For this receptor, the in vitro activity estimates (pEC50) in the absence of HSA are comparable for all peptides because they reflect the albumin-independent binding (free peptide) value (0% HSA, Table 8). However, in the presence of albumin (BSA or HSA), the activity measurements via the acylated peptides are not comparable (Table 8). This is due to the variability in the albumin-acylpeptide binding efficiency, which depends collectively on the amino acid sequence, acylating motif, attachment site, and linker length of each peptide. For this conjugated polypeptide, however, the decrease in activity (decrease in pEC50 value) in the presence compared to the absence of HSA qualitatively indicates an albumin-peptide interaction (Table 8, Figs. 2A and 2B). In contrast, linear polypeptides (pramlintide, human calcitonin) are not affected by the presence of HSA (Table 8, Figs. 2C and 2D), reflecting their poor albumin binding efficiency.

Example 8: Intravenous infusion of “linear” (i.e., non-acylated) amylin analog polypeptides: Pharmacokinetic studies to assess renal clearance (CL) of linear amylin analog polypeptides

The peptides were dissolved in sterile saline and administered as a 3-hour intravenous infusion in non-fasted male Sprague-Dawley rats (n=3 per group) via a femoral vein cannula at a final dose of 0.100 mg/kg. The drugs were infused at a rate of 1.67 ml/kg/h. Blood samples (approximately 250 μl) were collected for pharmacokinetic analysis via a jugular vein cannula at 0.25, 0.5, 1, 2, 3, 3.17, 3.33, 3.5, 4, 4.5, 5, and 6 h after the start of the infusion (long-term method) or at 1, 1.5, 2, 2.5, and 3 h after the start of the infusion (resting-state screening method). All samples were collected in Microtainerb tubes containing K2EDTA as an anticoagulant and 25 μl of protease inhibitor. Plasma was obtained by centrifugation and stored at -80°C until analysis. The results of this analysis are shown in Table 9 and some copies in Figs. 3a-3b.

Example 9: Intravenous infusion of “conjugated” (i.e., acylated) amylin analog polypeptides: Pharmacokinetic studies to assess renal clearance (CL) of conjugated amylin analog polypeptides

Peptides were dissolved in sterile saline and administered as a 1-hour intravenous infusion in non-fasted male Sprague-Dawley rats (n=3 per group) via a femoral vein cannula at a final dose of 0.033 mg/kg. Drugs were infused at a rate of 1.67 ml/kg/h. Blood samples (approximately 250 μl) were collected for pharmacokinetic analysis via a jugular vein cannula at 0.25, 0.5, 0.75, 1, 1.17, 1.33, 1.5, 2, 4, 6, 8, 24, 30, and 48 h after the start of infusion into Microtainer tubes containing K2EDTA as an anticoagulant and 25 μl of protease inhibitor. Plasma was obtained by centrifugation and stored at -80°C until analysis. The results of this analysis are shown in Table 9 and several copies in Fig. 3a-3b.

Example 10: Subcutaneous Infusion: Pharmacokinetic Studies to Assess Renal Clearance (CL) of Amylin Analogue Polypeptides

The peptides were dissolved in sterile saline and administered as a 1-hour subcutaneous infusion to non-fasted male Sprague-Dawley rats (n=3 per group) at a final dose of 0.033 mg/kg via a cannula placed in the subcutaneous space between the scapulae. The drugs were infused at a rate of 0.145 ml/h/kg. Blood samples (approximately 250 μl) were collected for pharmacokinetic analysis via a jugular venous cannula at 0.25, 0.5, 1, 1.5, 2, 4, 6, 8, 24, 30, and 48 h after the start of the infusion into Microtainer tubes containing K2EDTA as an anticoagulant and 25 μl of protease inhibitor. Plasma was obtained by centrifugation and stored at -80°C until analysis. The results of this analysis are presented in Table 9.

Example 11: Subcutaneous Bolus Injection: Pharmacokinetic Studies to Assess Renal Clearance (CL) of Amylin Analogue Polypeptides

The peptides were dissolved in sterile saline and administered to non-fasted male Sprague-Dawley rats (n=3 per group) at a dose of 0.3 mg/kg via a bolus injection placed subcutaneously between the scapulae. Blood samples (approximately 250 μL) were collected for pharmacokinetic analysis via a jugular venous cannula at 0.083, 0.167, 0.25, 0.5, 1, 2, 4, 8, 24, 30, and 48 h post-dose in Microtainer tubes containing K2EDTA as an anticoagulant and 25 μL of protease inhibitor. Plasma was obtained by centrifugation and stored at -80°C until analysis. The results of this analysis are presented in Table 9.

Example 12: Method for preparing a plasma sample for pharmacokinetic studies

Protein precipitation

A 60 μl aliquot of each plasma sample was placed into a 96-well plate. To each well, 6 μl of 0.5% Tween-20 was added. The plates were then vortexed for 10 min at 1200 rpm before the appropriate internal standard, containing 180 μl of 0.1% TFA in 2:1 ethanol:acetonitrile, was added to each well. The plates were vortexed for 5 min at 1300 rpm and then centrifuged for 10 min at 2844 x g. The supernatants (180 μl) were placed into a fresh 96-well plate and evaporated under a stream of nitrogen at 45°C. The residues were reconstituted in 80 μl of 20% acetonitrile (aq) containing 0.1% formic acid.

Solid phase extraction

A 60-mL aliquot of each plasma sample was diluted with 180 mL of 10 mM ammonium acetate (pH 6.8) containing the appropriate internal standard and loaded into an Oasis WCX microelution plate (Waters Corporation, Milford, MA) that had been pretreated with 200 mL of methanol and 200 mL of deionized water. Samples were washed with 200 mL of 5% ammonium hydroxide (aq) and then with 200 mL of 20% acetonitrile in water. The analyte was eluted with 200 mL of 5% formic acid in 75:25 acetonitrile:water. The eluent was dried under a stream of nitrogen. The residues were reconstituted in 80 μL of 20% acetonitrile (aq) containing 0.1% formic acid.

Example 13: Quantitative LC/MS analysis of amylin analogue polypeptides in plasma

All calibration standards were prepared in control rat plasma containing a mixture of K2EDTA and protease inhibitor.

Samples and standards were analyzed by TurboIonSpray™ UPLC-MS/MS using a system consisting of a CTC HTS PAL autoinjector (Leap, Carrboro, NC), an Agilent Infinity 1290 system with column oven (Palo Alto, CA), a Valeo switching valve (Houston, TX), and a Sciex API 5600 TripleTOF™ or Sciex API 4000QTrap AB mass spectrometer (Framingham, MA). Samples were injected onto a 2.1 x 50 mm C18 reversed-phase analytical column, typically a Waters CORTECS UPLC C18+, 1.6 μM (Waters Corporation, Milford, MA) or equivalent. Chromatographic separation was achieved by a gradient method using water containing 0.1% formic acid (A) and acetonitrile containing 0.1% formic acid (B) as the mobile phase. Initial conditions consisted of 90% A and 10% B. The organic component was increased to 95% B over 3-1 min depending on the peptide. Typical flow rates were 600 μl/min. The column temperature was maintained constant at 40 or 50°C. Peptides were quantified by monitoring one or more product ions derived from the multiply charged parent ion.

Example 14: In vivo efficacy of amylin analog polypeptides in inhibiting food intake in rats

Acute food intake was measured continuously for 72 hours using the BioDAQ Food Monitoring System (Research Diets, New Brunswick, NJ) to quantify the inhibition of food intake by these amylin analog polypeptides. Long-Evans rats, approximately 8 weeks of age, were used. Rats were housed individually and acclimated to a 45% fat diet for at least 2 weeks prior to dosing. After 1 week of acclimation, all rats were housed individually in BioDAQ cages (Research Diets, New Brunswick, NJ) and maintained at a constant temperature (approximately 22°C) and 30-70% relative humidity with a 12-hour light/dark cycle (lights on 7:00 AM to 7:00 PM). Rats were given ad libitum access to water and pelleted chow (Research Diets D12451i, 45 kcal % fat, Research Diets, New Brunswick, NJ). All procedures were in accordance with the Animal Welfare Act, USDA regulations, and approved by the Mispro Institutional Animal Care and Use Committee. Animals were randomized to treatment groups based on body weight (N=8 rats/group). Animals were dosed (SC bolus injection) with the indicated concentration of amylin analog polypeptide or placebo (saline) from 6:00 to 6:30 AM before lights on in closed chambers during dosing. Chambers were opened and continuous data collection began immediately after dosing was completed. Data were analyzed using BioDAQ Viewer software (version 2.3.07), and filters were used as needed to reduce noise in the data due to adverse feeding behavior. All data are expressed as % inhibition relative to placebo and are presented as mean. Data were analyzed for statistical significance using Microsoft Excel (Redmond, WA) with a 2-sample t-test. Values of P<0.05 were considered to indicate a significant difference between treatment groups. % inhibition of acute food intake relative to placebo for the amylin analog polypeptides are shown in Table 10.

Example 15: In vivo efficacy on body weight in LE DIO rats after 13 days

Chronic (13 days) in vivo dose-response efficacy studies were conducted in a rodent model of obesity (diet-induced obese Long-Evans (DIO) rats) to examine the efficacy and durability of amylin analog polypeptides on weight loss. Male LE DIO rats (Envigo Laboratories, Inc., Indianapolis, IN) were used and were fed a high-fat diet (Teklad TD 95217, 40% kcal from fat, Harlan Laboratories, Madison, WI) from weaning. Rats were 15-17 weeks old at the start of the study. Rats were housed one per cage, had ad libitum access to a high-fat diet (Harlan TD.95217, 4.3 kcal/g) and water, were maintained on a 12-h light/dark cycle from 5:00 a.m. to 5:00 p.m. at 21°C and 50% relative humidity, and were allowed to acclimate for at least 10 days prior to surgery. All procedures were performed in accordance with the Animal Welfare Act, USDA regulations, and were approved by the Mispro Institutional Animal Care and Use Committee. Body weight measurements were performed twice weekly beginning 3 days prior to surgery. Baseline measurements of adipose tissue mass and non-adipose tissue mass were taken 3 days prior to the start of peptide infusion using a QMR instrument (Echo Medical Systems, Houston, TX). Rats were randomized to different treatment groups (n=4–6 rats/group) according to body fat percentage and/or body weight. Alzet miniature osmotic pumps (2 weeks; Model 2002, Durect Corporation, Cupertino, CA) were filled under sterile conditions with vehicle or peptide one day before surgery. On the day of surgery, rats were anesthetized under isoflurane and the dorsal skin surface was shaved and cleaned. Rats were injected subcutaneously with flunexin (2.5 mg/kg). A 1–2 cm surgical incision was made between the scapulae. A 2–3 cm subcutaneous tunnel was created by blunt dissection, into which a sterile, filled osmotic miniature pump was inserted. The skin incision was closed with skin clips. Each rat was implanted with one or two osmotic pumps containing vehicle or peptide according to their treatment group. Data were analyzed in Excel and/or Prism (GraphPad Software, Inc., La Jolla, CA) using one-way analysis of variance to compare each group with its corresponding control group. Values of P<0.05 were considered to indicate a significant difference between treatment groups. Mean weight loss (%) from baseline and placebo (ΔΔ) from the 13-day studies are shown in Table 9.

Example 16: Weight Loss Efficacy of Amylin Analogue Polypeptides in Combination with Exenatide in LE DIO Rats

A chronic study was conducted to determine the effects and durability of continuous administration of amylin analog polypeptides in combination with exenatide (a GLP-1 receptor agonist) on body weight after 27 days of treatment in LE DIO rats. Male 18-week-old LE DIO rats (14 weeks on a high-fat diet) were implanted either subcutaneously (s.c.) with two (2) Alzet osmotic minipumps containing the indicated doses of amylin analog polypeptide and/or exenatide (10 μg/kg/d = ED50 for weight loss) or placebo (20% DMSO in water) (N = 8 animals/treatment group). Amylin analog polypeptides, whose RI was maintained by q2d administration, were administered via q2d injection instead of minipump administration. All other procedures were the same as described for the previous example. Mean weight loss (%) from baseline and placebo (ΔΔ) obtained from chronic combination studies with exenatide are shown in Table 11.

Example 17: Antidiabetic efficacy induced by amylin analogue polypeptides in combination with exenatide in ZDF rats

A chronic study was conducted to determine the antidiabetic effects of continuous administration of an amylin analog polypeptide in combination with exenatide on HbA1c (the primary antidiabetic parameter) after 27 days of treatment in Zucker diabetic (ZDF) rats. Male ZDF rats were obtained at six (6) weeks of age (Charles River, Raleigh, NC) and used in the study at eight (8) weeks of age. At the start of the study, rats were housed one per cage with ad libitum access to Purina 5008 chow (Lab Diet, St. Louis, MO) and water, maintained on a 12 h light/dark cycle from 5:00 AM to 5:00 PM at 21°C and 50% relative humidity, and allowed to acclimate for (9) days prior to study initiation. A preliminary blood draw (day -3) was performed via the tail vein for glucose and HbA1c measurements. ZDF rats with similar mean HbA1c and glucose were randomized to treatment groups (n=10/group) and implanted subcutaneously (s.c.) with two (2) Alzet osmotic minipumps containing the indicated doses of amylin analog polypeptide and/or exenatide (10 μg/kg/day) or placebo (20% DMSO in water) (n=10 animals/treatment group). Amylin analog polypeptides, whose RC was maintained by administration every two days (qdd), were administered via qdd s.c. injection instead of minipump insertion. All other procedures were the same as described for the previous example. Blood samples were repeated on days 14 and 27 (end of study) for glucose and HbA1c measurements. Final whole blood samples were collected by cardiac puncture under isoflurane anesthesia (day 27). HbA1c analysis was performed using a Carolina Chemistries CLC720i clinical chemistry analyzer (Mindray Inc., Mahwah, NJ) with the protocol and method parameters described by the manufacturer. HbAlc results expressed as % mean change from baseline and placebo (ΔΔ) obtained from chronic combination studies with exenatide are shown in Table 11.

Although the present invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not to limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages and modifications are within the scope of the following claims.

Other embodiments

Although the present invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not to limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages and modifications are within the scope of the following claims.

--->

SEQUENCE LIST

<110> INTARCIA THERAPEUTICS, INC.

<120> HUMAN AMYLIN ANALOGUE POLYPEPTIDES AND METHODS OF THEIR APPLICATION

<130> 616782:102487-055PC

<140>

<141>

<150> 62/744236

<151> 11.10.2018

<160> 318

<170> PatentIn version 3.5

<210> 1

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 1

His Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 2

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 2

Ile Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 3

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> Lys(CO(CH2)4CO2H)

<400> 3

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 4

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 4

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ser Asn Phe Leu

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 5

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(gammaGlu-dpeg-dpeg-(gammaGlu)4-CO(CH2)14CH3)

<400> 5

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 6

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Leu

<400> 6

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 7

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 7

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Lys Thr Lys Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 8

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 8

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Lys Thr Tyr

35

<210> 9

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 9

Ser Cys Asn Thr Ala Ser Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu

1 5 10 15

Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 10

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (17)..(17)

<223> Lys(gammaGlu-CO(CH2)14CH3)

<400> 10

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

Xaa Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 11

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 11

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Arg Thr Lys Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 12

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(dpeg-dpeg-gammaGlu-CO(CH2)14CH3)

<400> 12

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 13

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)3-CO(CH2)16CH3)

<400> 13

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 14

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys((gammaGlu)5-CO(CH2)14CH3)

<400> 14

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 15

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 15

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Tyr Leu

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 16

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(dpeg-dpeg-gammaGlu-CO(CH2)14CH3)

<400> 16

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Gly Thr Tyr

35

<210> 17

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 17

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 18

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> Lys(CO(CH2)6CO2H)

<400> 18

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 19

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(gammaGlu-dpeg-dpeg-(gammaGlu)3-CO(CH2)14CH3)

<400> 19

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 20

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(dpeg-dpeg-gammaGlu-CO(CH2)14CH3)

<400> 20

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Ala Thr Tyr

35

<210> 21

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys((gammaGlu)4-CO(CH2)14CH3)

<400> 21

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 22

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 22

Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Tyr Leu

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 23

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 23

Ser Cys Ser Thr Ala Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu

1 5 10 15

Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 24

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 24

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Ile Leu

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 25

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(gammaGlu-dpeg-dpeg-(gammaGlu)-CO(CH2)14CH3)

<400> 25

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 26

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(gammaGlu-dpeg-dpeg-(gammaGlu)2-CO(CH2)14CH3)

<400> 26

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 27

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)2-CO(CH2)14CH3)

<400> 27

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 28

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)2-CO(CH2)14CH3)

<400> 28

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Gln Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 29

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys((gammaGlu)3-CO(CH2)14CH3)

<400> 29

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 30

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 30

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Glu Asn Phe Leu

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 31

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> Lys(CO(CH2)11CO2H)

<400> 31

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 32

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 32

Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Glu Leu

1 5 10 15

Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 33

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 33

Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ser Asn Glu Leu

1 5 10 15

His Arg Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 34

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 34

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ser Asn Glu Leu

1 5 10 15

His Arg Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 35

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Leu

<400> 35

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Asn Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 36

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(gammaGlu-CO(CH2)14CH3)

<400> 36

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 37

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 37

Ser Cys Asn Thr Ala Ser Cys Ala Thr Gln Arg Leu Ala Asn Tyr Leu

1 5 10 15

Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 38

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 38

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 39

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)2-Gly-CO(CH2)14CH3)

<400> 39

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 40

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 40

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr His Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 41

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 41

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Arg Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Asn Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 42

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> Lys(gammaGlu-CO(CH2)14CH3)

<400> 42

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 43

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 43

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 44

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 44

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 45

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 45

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Arg Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 46

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Leu

<400> 46

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa

1 5 10 15

Val Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 47

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(gammaGlu-CO(CH2)14CH3)

<400> 47

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 48

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(dpeg-dpeg- gammaGlu-(CO(CH2)16CO2H))

<400> 48

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Gln Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 49

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 49

Ser Cys Ser Thr Ala Thr Cys Ala Thr Gln Arg Leu Ala Asn Tyr Leu

1 5 10 15

Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 50

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Leu

<400> 50

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 51

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 51

Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Glu Leu

1 5 10 15

Val Arg Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 52

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 52

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ser Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 53

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)3-CO(CH2)14CH3)

<400> 53

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 54

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 54

Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Glu Leu

1 5 10 15

His Arg Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 55

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(CO(CH2)16CO2H))

<400> 55

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 56

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)2-CO(CH2)16CH3)

<400> 56

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 57

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 57

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 58

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 58

Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 59

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 59

Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Tyr Leu

1 5 10 15

Gln Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 60

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> D-Phe

<400> 60

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 61

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 61

Ser Cys Asn Thr Ala Ser Cys Ala Thr Gln Arg Leu Ala Asn Tyr Leu

1 5 10 15

His Arg Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 62

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 62

Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 63

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(gammaGlu-dpeg-dpeg-gammaGlu-CO(CH2)14CH3)

<400> 63

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 64

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys((gammaGlu)2(CO(CH2)18CO2H))

<400> 64

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 65

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys((gammaGlu)2(CO(CH2)16CO2H))

<400> 65

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 66

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)4-CO(CH2)14CH3)

<400> 66

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 67

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)3-CO(CH2)14CH3)

<400> 67

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 68

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> Lys((gammaGlu)2-CO(CH2)14CH3)

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(2(gammaGlu)-CO(CH2)14CH3)

<400> 68

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 69

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(dpeg-dpeg-gammaGlu-CO(CH2)14CH3)

<400> 69

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Tyr Thr Tyr

35

<210> 70

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)2-CO(CH2)14CH3)

<400> 70

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 71

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)2-dpeg-dpeg- gammaGlu-CO(CH2)14CH3)

<400> 71

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 72

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> Lys(CO(CH2)14CO2H))

<400> 72

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 73

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(gammaGlu-CO(CH2)18CO2H)

<400> 73

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 74

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(CO(CH2)10CO2H))

<400> 74

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 75

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(CO(CH2)18CO2H)

<400> 75

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 76

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(gammaGlu-CO(CH2)16CO2H)

<400> 76

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 77

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)3-Gly-CO(CH2)14CH3)

<400> 77

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 78

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(dpeg-dpeg-gammaGlu-CO(CH2)14CH3)

<400> 78

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Pro Thr Tyr

35

<210> 79

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(CO(CH2)15CO2H)

<400> 79

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 80

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(gammaGlu-CO(CH2)15CO2H)

<400> 80

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 81

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(CO(CH2)12CO2H)

<400> 81

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 82

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> Lys((gammaGlu)2-CO(CH2)14CO2H)

<400> 82

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 83

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> Lys(CO(CH2)16CO2H)

<400> 83

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 84

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(CO(CH2)8CO2H)

<400> 84

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 85

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> Lys((gammaGlu)2-dpeg-dpeg-gammaGlu-CO(CH2)18CO2H)

<400> 85

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Tyr Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 86

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(CO(CH2)14CO2H)

<400> 86

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 87

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> Lys((gammaGlu)2-CO(CH2)16CO2H)

<400> 87

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 88

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(gammaGlu-CO(CH2)14CO2H)

<400> 88

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 89

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> Lys(CO(CH2)18CO2H)

<400> 89

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 90

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(CO(CH2)6CO2H)

<400> 90

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 91

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)5-CO(CH2)14CH3)

<400> 91

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 92

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> Lys((gammaGlu)2-CO(CH2)18CO2H)

<400> 92

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 93

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(CO(CH2)16CO2H)

<400> 93

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 94

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> Lys((gammaGlu)2-dpeg-dpeg-gammaGlu-CO(CH2)18CO2H)

<400> 94

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 95

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> D-Lys(CO(CH2)18CO2H)

<400> 95

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 96

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> D-Lys((gammaGlu)2-dpeg-CO(CH2)18CO2H)

<400> 96

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 97

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> D-Lys((gammaGlu)2-CO(CH2)18CO2H)

<400> 97

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 98

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> Lys(dpeg-dpeg- gammaGlu-CO(CH2)16CO2H)

<400> 98

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 99

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (17)..(17)

<223> Lys(dpeg-dpeg- gammaGlu-CO(CH2)16CO2H)

<400> 99

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

Xaa Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 100

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> D-Lys((gammaGlu)2-dpeg-gammaGlu-CO(CH2)18CO2H)

<400> 100

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 101

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> D-Lys((gammaGlu)2-dpeg-dpeg- gammaGlu-CO(CH2)18CO2H)

<400> 101

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 102

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(dpeg-dpeg-gammaGlu-CO(CH2)16CO2H)

<400> 102

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 103

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)2-dpeg-dpeg- gammaGlu-CO(CH2)18CO2H)

<400> 103

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 104

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)3-CO(CH2)16CO2H)

<400> 104

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 105

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(dpeg- gammaGlu-CO(CH2)16CO2H)

<400> 105

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Gln Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 106

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(gammaGlu-CO(CH2)16CO2H)

<400> 106

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Gln Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 107

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (13)..(13)

<223> Lys(gammaGlu-CO(CH2)14CH3)

<400> 107

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Xaa Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 108

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (8)..(8)

<223> Lys(gammaGlu-CO(CH2)14CH3)

<400> 108

Ser Cys Asn Thr Ser Thr Cys Xaa Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 109

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(gammaGlu-CO(CH2)16CO2H)

<400> 109

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 110

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(gammaGlu-CO(CH2)16CO2H)

<400> 110

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Tyr Leu

1 5 10 15

Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 111

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 111

Ser Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Tyr Leu

1 5 10 15

Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 112

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys((gammaGlu)2-dpeg-dpeg-gammaGlu-CO(CH2)18CO2H)

<400> 112

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 113

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(dpeg-dpeg- gammaGlu-CO(CH2)16CO2H)

<400> 113

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 114

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys((gammaGlu)2-CO(CH2)18CO2H)

<400> 114

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 115

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(CO(CH2)18CO2H)

<400> 115

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 116

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(gammaGlu-CO(CH2)18CO2H)

<400> 116

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 117

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys((gammaGlu)2-CO(CH2)10CO2H)

<400> 117

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 118

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys((gammaGlu)2-CO(CH2)6CO2H)

<400> 118

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 119

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(gammaGlu)-CO(CH2)18CO2H)

<400> 119

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 120

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(CO(CH2)16CO2H)

<400> 120

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 121

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (12)..(12)

<223> Lys(gammaGlu)-CO(CH2)14CH3)

<400> 121

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Xaa Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 122

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys((gammaGlu)2-dpeg-dpeg-gammaGlu-CO(CH2)14CH3)

<400> 122

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 123

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys((gammaGlu)2-CO(CH2)14CH3)

<400> 123

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 124

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)2-CO(CH2)20CH3)

<400> 124

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 125

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys(gammaGlu-CO(CH2)14CH3)

<400> 125

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 126

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(dpeg-dpeg- gammaGlu-CO(CH2)16CO2H)

<400> 126

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 127

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys

<400> 127

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 128

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 128

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Lys

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 129

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys

<400> 129

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 130

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 130

Lys Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 131

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 131

Lys Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 132

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (15)..(15)

<223> Aib

<400> 132

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Xaa Leu

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 133

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (19)..(19)

<223> Aib

<400> 133

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu

1 5 10 15

Val Lys Xaa Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 134

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Aib

<400> 134

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 135

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (17)..(17)

<223> Aib

<400> 135

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu

1 5 10 15

Xaa Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 136

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> N-MeLeu

<400> 136

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ser Asn Phe Xaa

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 137

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> N-MeLeu

<400> 137

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Xaa

1 5 10 15

Val Lys Ser Ser Asn Glu Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 138

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (14)..(14)

<223> D-Asn

<220>

<221>MOD_RES

<222> (16)..(16)

<223> Lys((gammaGlu)5-(CO(CH2)14CH3)

<400> 138

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Xaa Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Gly Thr Pro

35

<210> 139

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((gammaGlu)2-(CO(CH2)18CO2H)

<400> 139

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Pro

35

<210> 140

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys((gammaGlu)2-(CO(CH2)16CO2H)

<220>

<221>MOD_RES

<222> (14)..(14)

<223> D-Asn

<400> 140

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Xaa Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Gly Thr Pro

35

<210> 141

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys((gammaGlu)2-(CO(CH2)16CO2H)

<220>

<221>MOD_RES

<222> (14)..(14)

<223> D-Asp

<400> 141

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Xaa Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Gly Thr Pro

35

<210> 142

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys((gammaGlu)2-(CO(CH2)16CO2H)

<220>

<221>MOD_RES

<222> (14)..(14)

<223> D-Asp

<400> 142

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Xaa Glu Leu

1 5 10 15

Arg His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Gly Thr Pro

35

<210> 143

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys((gammaGlu)2-(CO(CH2)14CO2H)

<400> 143

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 144

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(acetyl)

<400> 144

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 145

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(allyloxycarbonyl)

<400> 145

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 146

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys((dimethyl-2,6-di-oxocyclohex-1-ylene)ethyl)

<400> 146

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 147

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(gammaGlu)

<400> 147

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 148

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(gammaGlu-acetyl)

<400> 148

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 149

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(gammaGlu-trityl)

<400> 149

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 150

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(gammaGlu-tert-butyl)

<400> 150

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 151

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(gammaGlu-gammaGlu)

<400> 151

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 152

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(gammaGlu-gammaGlu-acetyl)

<400> 152

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 153

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(gammaGlu-gammaGlu-trityl)

<400> 153

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 154

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (16)..(16)

<223> D-Lys(gammaGlu-gammaGlu-tert-butyl)

<400> 154

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Xaa

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 155

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 155

Lys Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 156

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(Fmoc)

<400> 156

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 157

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(gammaGlu)

<400> 157

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 158

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(gammaGlu-acetyl)

<400> 158

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 159

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(gammaGlu-trityl)

<400> 159

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 160

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(gammaGlu-tert-butyl)

<400> 160

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 161

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(gammaGlu-gammaGlu)

<400> 161

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 162

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(gammaGlu-gammaGlu-acetyl)

<400> 162

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 163

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(gammaGlu-gammaGlu-trityl)

<400> 163

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 164

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(gammaGlu-gammaGlu-tert-butyl)

<400> 164

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Ser Arg Leu Ala Asn Phe Leu

1 5 10 15

Gln Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 165

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(gammaGlu)

<400> 165

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 166

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Lys(gammaGlu-gammaGlu)

<400> 166

Xaa Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 167

<400> 167

000

<210> 168

<400> 168

000

<210> 169

<400> 169

000

<210> 170

<400> 170

000

<210> 171

<400> 171

000

<210> 172

<400> 172

000

<210> 173

<400> 173

000

<210> 174

<400> 174

000

<210> 175

<400> 175

000

<210> 176

<400> 176

000

<210> 177

<400> 177

000

<210> 178

<400> 178

000

<210> 179

<400> 179

000

<210> 180

<400> 180

000

<210> 181

<400> 181

000

<210> 182

<400> 182

000

<210> 183

<400> 183

000

<210> 184

<400> 184

000

<210> 185

<400> 185

000

<210> 186

<400> 186

000

<210> 187

<400> 187

000

<210> 188

<400> 188

000

<210> 189

<400> 189

000

<210> 190

<400> 190

000

<210> 191

<400> 191

000

<210> 192

<400> 192

000

<210> 193

<400> 193

000

<210> 194

<400> 194

000

<210> 195

<400> 195

000

<210> 196

<400> 196

000

<210> 197

<400> 197

000

<210> 198

<400> 198

000

<210> 199

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace="Lys", or "D-Lys", or "His", or "Ile"

<220>

<221> OPTION

<222> (3)..(3)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (5)..(5)

<223> /replace=«Ala»

<220>

<221> OPTION

<222> (6)..(6)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (8)..(8)

<223> /replace=«Lys»

<220>

<221> OPTION

<222> (10)..(10)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (12)..(12)

<223> /replace=«Lys»

<220>

<221> OPTION

<222> (13)..(13)

<223> /replace="Ser", or "Glu", or "Lys"

<220>

<221> OPTION

<222> (14)..(14)

<223> /replace="D-Asn" or "D-Asp" or "Tyr" or "Gln"

<220>

<221> OPTION

<222> (15)..(15)

<223> /replace=«Phe» or «D-Phe» or «Tyr» or «Ile» or «D-Lys» or «Lys»

or "alpha-aminoisobutyric acid (Aib)"

<220>

<221> OPTION

<222> (16)..(16)

<223> /replace= «D-Lys», or «Leu», or «Aib», or «N-methyl leucine

(N-MeL)" or "D-Leu"

<220>

<221> OPTION

<222> (17)..(17)

<223> /replace="Val" or "Gln" or "Arg" or "D-Lys" or "Lys" or "Aib"

<220>

<221> OPTION

<222> (18)..(18)

<223> /replace="His", or "Arg"

<220>

<221> OPTION

<222> (19)..(19)

<223> /replace=«Aib»

<220>

<221> OPTION

<222> (20)..(20)

<223> /replace=«Aib»

<220>

<221> OPTION

<222> (22)..(22)

<223> /replace=«Glu»

<220>

<221> OPTION

<222> (29)..(29)

<223> /replace="Arg" or "Lys"

<220>

<221> OPTION

<222> (31)..(31)

<223> /replace="D-Lys", or "Asn", or "His"

<220>

<221> OPTION

<222> (35)..(35)

<223> /replace="D-Glu" or "Asn" or "Lys" or "Gly" or "Ala" or "Tyr" or "Pro"

<220>

<221> OPTION

<222> (37)..(37)

<223> /replace=«Pro»

<220>

<221> SITE

<222> (1).(37)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 199

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Lys

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 200

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace="Lys", or "D-Lys", or "His", or "Ile"

<220>

<221> OPTION

<222> (3)..(3)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (5)..(5)

<223> /replace=«Ala»

<220>

<221> OPTION

<222> (6)..(6)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (8)..(8)

<223> /replace=«Lys»

<220>

<221> OPTION

<222> (10)..(10)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (12)..(12)

<223> /replace=«Lys»

<220>

<221> OPTION

<222> (13)..(13)

<223> /replace="Ser", or "Glu", or "Lys"

<220>

<221> OPTION

<222> (14)..(14)

<223> /replace="D-Asn" or "D-Asp" or "Tyr" or "Gln"

<220>

<221> OPTION

<222> (15)..(15)

<223> /replace=«Phe» or «D-Phe» or «Tyr» or «Ile» or «D-Lys» or «Lys»

or "alpha-aminoisobutyric acid (Aib)"

<220>

<221> OPTION

<222> (16)..(16)

<223> /replace= «D-Lys», or «Leu», or «Aib», or «N-methyl leucine

(N-MeL)" or "D-Leu"

<220>

<221> OPTION

<222> (17)..(17)

<223> /replace="Val" or "Gln" or "Arg" or "D-Lys" or "Lys" or "Aib"

<220>

<221> OPTION

<222> (18)..(18)

<223> /replace="His", or "Arg"

<220>

<221> OPTION

<222> (19)..(19)

<223> /replace=«Aib»

<220>

<221> OPTION

<222> (20)..(20)

<223> /replace=«Aib»

<220>

<221> OPTION

<222> (22)..(22)

<223> /replace=«Glu»

<220>

<221> OPTION

<222> (29)..(29)

<223> /replace="Arg" or "Lys"

<220>

<221> OPTION

<222> (31)..(31)

<223> /replace= «D-Lys» or «Asn»

<220>

<221> OPTION

<222> (35)..(35)

<223> /replace= "D-Glu" or "Asn"

<220>

<221> SITE

<222> (1).(37)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 200

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Lys

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 201

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace="Lys", or "D-Lys", or "His", or "Ile"

<220>

<221> OPTION

<222> (3)..(3)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (5)..(5)

<223> /replace=«Ala»

<220>

<221> OPTION

<222> (6)..(6)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (8)..(8)

<223> /replace=«Lys»

<220>

<221> OPTION

<222> (10)..(10)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (12)..(12)

<223> /replace=«Lys»

<220>

<221> OPTION

<222> (13)..(13)

<223> /replace="Ser", or "Glu", or "Lys"

<220>

<221> OPTION

<222> (14)..(14)

<223> /replace="D-Asn" or "D-Asp" or "Tyr" or "Gln"

<220>

<221> OPTION

<222> (15)..(15)

<223> /replace=«Phe», or «D-Phe», or «Tyr», or «Ile», or «D-Lys»,

or "Lys" or "Aib"

<220>

<221> OPTION

<222> (16)..(16)

<223> /replace="D-Lys" or "Leu" or "Aib" or "N-MeL" or "D-Leu"

<220>

<221> OPTION

<222> (17)..(17)

<223> /replace="Val" or "Gln" or "Arg" or "D-Lys" or "Lys" or "Aib"

<220>

<221> OPTION

<222> (18)..(18)

<223> /replace="His", or "Arg"

<220>

<221> OPTION

<222> (19)..(19)

<223> /replace=«Aib»

<220>

<221> OPTION

<222> (20)..(20)

<223> /replace=«Aib»

<220>

<221> OPTION

<222> (22)..(22)

<223> /replace=«Glu»

<220>

<221> OPTION

<222> (29)..(29)

<223> /replace="Arg" or "Lys"

<220>

<221> SITE

<222> (1).(37)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 201

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Lys

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 202

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace= «D-Lys» or «Lys»

<220>

<221> OPTION

<222> (5)..(5)

<223> /replace=«Ala»

<220>

<221> OPTION

<222> (10)..(10)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (15)..(15)

<223> /replace=«Phe»

<220>

<221> OPTION

<222> (16)..(16)

<223> /replace=“D-Lys” or “Leu”

<220>

<221> OPTION

<222> (17)..(17)

<223> /replace=«Val» or «Gln»

<220>

<221> OPTION

<222> (18)..(18)

<223> /replace="His", or "Arg"

<220>

<221> OPTION

<222> (31)..(31)

<223> /replace=«Asn»

<220>

<221> OPTION

<222> (35)..(35)

<223> /replace=«Asn»

<220>

<221> SITE

<222> (1).(37)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 202

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Lys

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 203

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace= «D-Lys» or «Lys»

<220>

<221> OPTION

<222> (5)..(5)

<223> /replace=«Ala»

<220>

<221> OPTION

<222> (10)..(10)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (15)..(15)

<223> /replace=«Phe»

<220>

<221> OPTION

<222> (16)..(16)

<223> /replace=“D-Lys” or “Leu”

<220>

<221> OPTION

<222> (17)..(17)

<223> /replace=«Val» or «Gln»

<220>

<221> OPTION

<222> (18)..(18)

<223> /replace="His", or "Arg"

<220>

<221> SITE

<222> (1).(37)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 203

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Lys

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 204

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace= «Lys» or»«D-Lys»

<220>

<221> OPTION

<222> (10)..(10)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (15)..(15)

<223> /replace=«Phe»

<220>

<221> OPTION

<222> (16)..(16)

<223> /replace= «Lys» or»«D-Lys»

<220>

<221> OPTION

<222> (17)..(17)

<223> /replace=«Val» or «Gln»

<220>

<221> OPTION

<222> (35)..(35)

<223> /replace=«Asn»

<220>

<221> SITE

<222> (1).(37)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 204

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 205

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace= «Lys» or»«D-Lys»

<220>

<221> OPTION

<222> (10)..(10)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (15)..(15)

<223> /replace=«Phe»

<220>

<221> OPTION

<222> (16)..(16)

<223> /replace= «Lys» or»«D-Lys»

<220>

<221> OPTION

<222> (17)..(17)

<223> /replace=«Val» or «Gln»

<220>

<221> SITE

<222> (1).(37)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 205

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 206

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221> OPTION

<222> (15)..(15)

<223> /replace=«Phe»

<220>

<221> OPTION

<222> (16)..(16)

<223> /replace= «Lys» or»«D-Lys»

<220>

<221> OPTION

<222> (17)..(17)

<223> /replace=«Val» or «Gln»

<220>

<221> OPTION

<222> (35)..(35)

<223> /replace=«Asn»

<220>

<221> SITE

<222> (1).(37)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 206

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 207

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221> OPTION

<222> (15)..(15)

<223> /replace=«Phe»

<220>

<221> OPTION

<222> (16)..(16)

<223> /replace= «Lys» or»«D-Lys»

<220>

<221> OPTION

<222> (17)..(17)

<223> /replace=«Val» or «Gln»

<220>

<221> SITE

<222> (1).(37)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 207

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 208

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace=«D-Lys»

<220>

<221> OPTION

<222> (10)..(10)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (15)..(15)

<223> /replace=«Phe»

<220>

<221> OPTION

<222> (16)..(16)

<223> /replace= «Lys» or»«D-Lys»

<220>

<221> OPTION

<222> (17)..(17)

<223> /replace=«Val» or «Gln»

<220>

<221> OPTION

<222> (35)..(35)

<223> /replace=«Asn»

<220>

<221> SITE

<222> (1).(37)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 208

Lys Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 209

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace=«D-Lys»

<220>

<221> OPTION

<222> (10)..(10)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (15)..(15)

<223> /replace=«Phe»

<220>

<221> OPTION

<222> (16)..(16)

<223> /replace= «Lys» or»«D-Lys»

<220>

<221> OPTION

<222> (17)..(17)

<223> /replace=«Val» or «Gln»

<220>

<221> SITE

<222> (1).(37)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 209

Lys Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 210

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace="Lys", or "D-Lys", or "His", or "Ile"

<220>

<221> OPTION

<222> (3)..(3)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (5)..(5)

<223> /replace=«Ala»

<220>

<221> OPTION

<222> (6)..(6)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (8)..(8)

<223> /replace=«Lys»

<220>

<221> OPTION

<222> (10)..(10)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (12)..(12)

<223> /replace=«Lys»

<220>

<221> OPTION

<222> (13)..(13)

<223> /replace=«Ser», or «Glu», or «Lys»

<220>

<221> OPTION

<222> (14)..(14)

<223> /replace="D-Asn" or "D-Asp" or "Tyr" or "Gln"

<220>

<221> OPTION

<222> (15)..(15)

<223> /replace=«Phe» or «D-Phe» or «Tyr» or «Ile» or «D-Lys» or «Lys»

or "alpha-aminoisobutyric acid (Aib)"

<220>

<221> OPTION

<222> (16)..(16)

<223> /replace= «D-Lys», or «Leu», or «Aib», or «N-methyl leucine

(N-MeL)" or "D-Leu"

<220>

<221> OPTION

<222> (17)..(17)

<223> /replace="Val" or "Gln" or "Arg" or "D-Lys" or "Lys" or "Aib"

<220>

<221> OPTION

<222> (18)..(18)

<223> /replace="His", or "Arg"

<220>

<221> OPTION

<222> (19)..(19)

<223> /replace=«Aib»

<220>

<221> OPTION

<222> (20)..(20)

<223> /replace=«Aib»

<220>

<221> OPTION

<222> (22)..(22)

<223> /replace=«Glu»

<220>

<221> OPTION

<222> (29)..(29)

<223> /replace="Arg" or "Lys"

<220>

<221> OPTION

<222> (31)..(31)

<223> /replace= «D-Lys» or «Asn»

<220>

<221> OPTION

<222> (35)..(35)

<223> /replace= "D-Glu" or "Asn"

<220>

<221> SITE

<222> (1).(37)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 210

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Lys

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 211

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace= «Lys» or»«D-Lys»

<220>

<221> OPTION

<222> (10)..(10)

<223> /replace=«Ser»

<220>

<221> OPTION

<222> (15)..(15)

<223> /replace=«Phe»

<220>

<221> OPTION

<222> (16)..(16)

<223> /replace= «Lys» or»«D-Lys»

<220>

<221> OPTION

<222> (17)..(17)

<223> /replace=«Val» or «Gln»

<220>

<221> OPTION

<222> (35)..(35)

<223> /replace=«Asn»

<220>

<221> SITE

<222> (1).(37)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 211

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 212

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<220>

<221> OPTION

<222> (15)..(15)

<223> /replace=«Phe»

<220>

<221> OPTION

<222> (16)..(16)

<223> /replace= «Lys» or»«D-Lys»

<220>

<221> OPTION

<222> (17)..(17)

<223> /replace=«Val» or «Gln»

<220>

<221> OPTION

<222> (35)..(35)

<223> /replace=«Asn»

<220>

<221> SITE

<222> (1).(37)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 212

Ser Cys Asn Thr Ser Thr Cys Ala Thr Gln Arg Leu Ala Asn Glu Leu

1 5 10 15

His Lys Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Lys Val

20 25 30

Gly Ser Glu Thr Tyr

35

<210> 213

<400> 213

000

<210> 214

<400> 214

000

<210> 215

<400> 215

000

<210> 216

<400> 216

000

<210> 217

<400> 217

000

<210> 218

<400> 218

000

<210> 219

<400> 219

000

<210> 220

<400> 220

000

<210> 221

<400> 221

000

<210> 222

<400> 222

000

<210> 223

<400> 223

000

<210> 224

<400> 224

000

<210> 225

<400> 225

000

<210> 226

<400> 226

000

<210> 227

<400> 227

000

<210> 228

<400> 228

000

<210> 229

<400> 229

000

<210> 230

<400> 230

000

<210> 231

<400> 231

000

<210> 232

<400> 232

000

<210> 233

<400> 233

000

<210> 234

<400> 234

000

<210> 235

<400> 235

000

<210> 236

<400> 236

000

<210> 237

<400> 237

000

<210> 238

<400> 238

000

<210> 239

<400> 239

000

<210> 240

<400> 240

000

<210> 241

<400> 241

000

<210> 242

<400> 242

000

<210> 243

<400> 243

000

<210> 244

<400> 244

000

<210> 245

<400> 245

000

<210> 246

<400> 246

000

<210> 247

<400> 247

000

<210> 248

<400> 248

000

<210> 249

<400> 249

000

<210> 250

<400> 250

000

<210> 251

<400> 251

000

<210> 252

<400> 252

000

<210> 253

<400> 253

000

<210> 254

<400> 254

000

<210> 255

<400> 255

000

<210> 256

<400> 256

000

<210> 257

<400> 257

000

<210> 258

<400> 258

000

<210> 259

<400> 259

000

<210> 260

<400> 260

000

<210> 261

<400> 261

000

<210> 262

<400> 262

000

<210> 263

<400> 263

000

<210> 264

<400> 264

000

<210> 265

<400> 265

000

<210> 266

<400> 266

000

<210> 267

<400> 267

000

<210> 268

<400> 268

000

<210> 269

<400> 269

000

<210> 270

<400> 270

000

<210> 271

<400> 271

000

<210> 272

<400> 272

000

<210> 273

<400> 273

000

<210> 274

<400> 274

000

<210> 275

<400> 275

000

<210> 276

<400> 276

000

<210> 277

<400> 277

000

<210> 278

<400> 278

000

<210> 279

<400> 279

000

<210> 280

<400> 280

000

<210> 281

<400> 281

000

<210> 282

<400> 282

000

<210> 283

<400> 283

000

<210> 284

<400> 284

000

<210> 285

<400> 285

000

<210> 286

<400> 286

000

<210> 287

<400> 287

000

<210> 288

<400> 288

000

<210> 289

<400> 289

000

<210> 290

<400> 290

000

<210> 291

<400> 291

000

<210> 292

<400> 292

000

<210> 293

<400> 293

000

<210> 294

<400> 294

000

<210> 295

<400> 295

000

<210> 296

<400> 296

000

<210> 297

<400> 297

000

<210> 298

<400> 298

000

<210> 299

<400> 299

000

<210> 300

<211> 37

<212> PROTEIN

<213> Homo sapiens

<400> 300

Lys Cys Asn Thr Ala Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu

1 5 10 15

Val His Ser Ser Asn Asn Phe Gly Ala Ile Leu Ser Ser Thr Asn Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 301

<211> 37

<212> PROTEIN

<213> Rattus sp.

<400> 301

Lys Cys Asn Thr Ala Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu

1 5 10 15

Val Arg Ser Ser Asn Asn Leu Gly Pro Val Leu Pro Pro Thr Asn Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 302

<211> 37

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 302

Lys Cys Asn Thr Ala Thr Cys Ala Thr Gln Arg Leu Ala Asn Phe Leu

1 5 10 15

Val His Ser Ser Asn Asn Phe Gly Pro Ile Leu Pro Pro Thr Asn Val

20 25 30

Gly Ser Asn Thr Tyr

35

<210> 303

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

polypeptide"

<400> 303

Lys Cys Asn Thr Ala Thr Cys Val Leu Gly Arg Leu Ser Gln Glu Leu

1 5 10 15

His Arg Leu Gln Thr Tyr Pro Arg Thr Asn Thr Gly Ser Asn Thr Tyr

20 25 30

<210> 304

<211> 32

<212> PROTEIN

<213> Homo sapiens

<400> 304

Cys Gly Asn Leu Ser Thr Cys Met Leu Gly Thr Tyr Thr Gln Asp Phe

1 5 10 15

Asn Lys Phe His Thr Phe Pro Gln Thr Ala Ile Gly Val Gly Ala Pro

20 25 30

<210> 305

<211> 32

<212> PROTEIN

<213> Unknown

<220>

<221> source

<223> /note=«Description of unknown:

salmon calcitonin»

<400> 305

Cys Ser Asn Leu Ser Thr Cys Val Leu Gly Lys Leu Ser Gln Glu Leu

1 5 10 15

His Lys Leu Gln Thr Tyr Pro Arg Thr Asn Thr Gly Ser Gly Thr Pro

20 25 30

<210> 306

<211> 37

<212> PROTEIN

<213> Unknown

<220>

<221> source

<223> /note=«Description of unknown:

beta-calcitonin gene-related peptide"

<400> 306

Ala Cys Asn Thr Ala Thr Cys Val Thr His Arg Leu Ala Gly Leu Leu

1 5 10 15

Ser Arg Ser Gly Gly Met Val Lys Ser Asn Phe Val Pro Thr Asn Val

20 25 30

Gly Ser Lys Ala Phe

35

<210> 307

<211> 39

<212> PROTEIN

<213> Unknown

<220>

<221> source

<223> /note=«Description of unknown:

exenatide sequence»

<400> 307

His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu

1 5 10 15

Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser

20 25 30

Ser Gly Ala Pro Pro Pro Ser

35

<210> 308

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

peptide"

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace= «D-Lys» or «Lys»

<220>

<221> OPTION

<222> (5)..(5)

<223> /replace=«Ala»

<220>

<221> SITE

<222> (1).(7)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 308

Ser Cys Asn Thr Ser Thr Cys

1 5

<210> 309

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

peptide"

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace= «Lys» or»«D-Lys»

<220>

<221> SITE

<222> (1).(7)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 309

SerCysAsnThrSerThrCys

1 5

<210> 310

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

peptide"

<400> 310

Ser Cys Asn Thr Ser Thr Cys

1 5

<210> 311

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

peptide"

<220>

<221>MOD_RES

<222> (1).(10)

<223> gammaGlu

<220>

<221> SITE

<222> (1).(10)

<223> /note="This sequence may span 1-10 residues"

<400> 311

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

1 5 10

<210> 312

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

peptide"

<220>

<221>MOD_RES

<222> (1).(10)

<223> gammaGlu

<220>

<221> SITE

<222> (1).(10)

<223> /note="This region may span 1-10 residues"

<400> 312

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly

1 5 10

<210> 313

<211> 4

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

peptide"

<220>

<221>MOD_RES

<222> (1).(4)

<223> gammaGlu

<400> 313

Xaa Xaa Xaa Xaa

1

<210> 314

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

peptide"

<220>

<221>MOD_RES

<222> (1).(5)

<223> gammaGlu

<400> 314

Xaa Xaa Xaa Xaa Xaa

1 5

<210> 315

<211> 4

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

peptide"

<220>

<221>MOD_RES

<222> (1).(3)

<223> gammaGlu

<400> 315

Xaa Xaa Xaa Gly

1

<210> 316

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

peptide"

<220>

<221>MOD_RES

<222> (2).(11)

<223> gammaGlu

<220>

<221> SITE

<222> (2).(11)

<223> /note="This region may span 1-10 residues"

<400> 316

Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

1 5 10

<210> 317

<211> 4

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note= «Description of the artificial sequence: Synthetic

peptide"

<400> 317

Glu Glu Glu Glu

1

<210> 318

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: Synthetic

peptide

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace=«D-Lys»

<220>

<221> SITE

<222> (1).(7)

<223> /note="The variant residues given in the sequence are not

preferred over those given in the footnotes

for alternative provisions"

<400> 318

LysCysAsnThrSerThrCys

1 5

<---

Claims (99)

1. An isolated polypeptide that is a functional analog of amylin, comprising the amino acid sequence of SEQ ID NO: 202: X ₁ CNTX ₅ TCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ X ₁₈ SSNNFGPILPPTX ₃₁ VGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 202), or a pharmaceutically acceptable salt thereof, wherein X ₁ represents S, k or K; X ₅ represents S; X ₁₀ represents Q or S; X ₁₅ represents E or F; X ₁₆ represents k, K or L; X ₁₇ represents H, V or Q; X ₁₈ represents K; X ₃₁ represents K or N; and X ₃₅ represents E or N; each K is independently L -lysine, optionally covalently linked to a lipophilic substituent, optionally through a spacer; each k is independently D -lysine, optionally covalently linked to a lipophilic substituent, optionally through a spacer; wherein two cysteine residues X ₁ CX ₃ TX ₅ X ₆ C are optionally additionally linked by a disulfide bridge; where L-amino acids are represented by capital single-letter designations and D-amino acids are represented by lowercase single-letter designations; provided that if X ₃₁ is N then X ₃₅ is E, or if X ₃₅ is N then X ₃₁ is K. 2. The isolated polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO: 204: X ₁ CNTSTCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 204), or a pharmaceutically acceptable salt thereof, wherein X ₁ represents S, K or k; X ₁₀ represents Q or S; X ₁₅ represents E or F; X ₁₆ represents L, K or k; X ₁₇ represents H, V, or Q; and X ₃₅ represents E or N; each K is independently L -lysine, optionally covalently linked to a lipophilic substituent, optionally through a spacer; each k is independently D -lysine, optionally covalently linked to a lipophilic substituent, optionally through a spacer; and wherein the two cysteine residues X ₁ CNTSTC (SEQ ID NO: 309) are optionally further linked by a disulfide bridge. 3. The isolated polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO: 206: SCNTSTCATQRLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSX ₃₅ TY-(OH/NH ₂ ) (SEQ ID NO: 206), or a pharmaceutically acceptable salt thereof, wherein X ₁₅ represents E or F; X ₁₆ represents L, K or k; X ₁₇ represents H, V, or Q; and X ₃₅ represents E or N; each K is independently L -lysine, optionally covalently linked to a lipophilic substituent, optionally through a spacer; each k is independently D -lysine, optionally covalently linked to a lipophilic substituent, optionally through a spacer; and wherein the two cysteine residues SCNTSTC (SEQ ID NO: 310) are optionally further linked by a disulfide bridge. 4. The isolated polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO: 209: X ₁ CNTSTCATX ₁₀ RLANX ₁₅ X ₁₆ X ₁₇ KSSNNFGPILPPTKVGSETY-(OH/NH ₂ ) (SEQ ID NO: 209), or a pharmaceutically acceptable salt thereof, wherein X ₁ represents K or k; X ₁₀ represents Q or S; X ₁₅ represents E or F; X ₁₆ represents L, K, or k; and X ₁₇ represents H, V or Q; each K is independently L -lysine, optionally covalently linked to a lipophilic substituent, optionally through a spacer; each k is independently D -lysine, optionally covalently linked to a lipophilic substituent, optionally through a spacer; and wherein the two cysteine residues X ₁ CNTSTC (SEQ ID NO: 318) are optionally further linked by a disulfide bridge. 5. An isolated polypeptide that is a functional analog of amylin, comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1-143, or a pharmaceutically acceptable salt thereof. 6. The isolated polypeptide according to claim 5, comprising the amino acid sequence: SC*NTSTC*ATQRLANEk*((γGlu) ₂ CO(CH ₂ ) ₁₄ CH ₃ )HKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 27), or a pharmaceutically acceptable salt thereof, where k* is D -lysine covalently linked to (γGlu) ₂ CO(CH ₂ ) ₁₄ CH ₃ ; and two cysteine residues, designated C*, at positions 2 and 7 are additionally linked by a disulfide bridge. 7. The isolated polypeptide according to claim 5, comprising the amino acid sequence: SC*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 57), or a pharmaceutically acceptable salt thereof, where two cysteine residues, designated C*, at positions 2 and 7 are additionally linked by a disulfide bridge. 8. The isolated polypeptide according to claim 5, comprising the amino acid sequence: K*((γGlu) _{2 (} CO( _CH2 ) _18CO2H ))C*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-( _NH2 ) (SEQ ID NO: 64), or a pharmaceutically acceptable salt thereof, where K* is L -lysine covalently linked to (γGlu) ₂ (CO(CH ₂ ) ₁₈ CO ₂ H); and two cysteine residues, designated C*, at positions 2 and 7 are additionally linked by a disulfide bridge. 9. The isolated polypeptide according to claim 5, comprising the amino acid sequence: K*((γGlu) _{2 (} CO( _CH2 ) _16CO2H ))C*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-( _NH2 ) (SEQ ID NO: 65), or a pharmaceutically acceptable salt thereof, where K* is L -lysine covalently linked to (γGlu) ₂ (CO(CH ₂ ) ₁₆ CO ₂ H); and two cysteine residues, designated C*, at positions 2 and 7 are additionally linked by a disulfide bridge. 10. The isolated polypeptide according to claim 5, comprising the amino acid sequence: K*(γGlu-CO(CH ₂ ) ₁₆ CO ₂ H)C*NTSTC*ATSRLANFLQKSSNNFGPILPPTKVGSETY-NH ₂ (SEQ ID NO: 109), or a pharmaceutically acceptable salt thereof, where K* is L -lysine covalently linked to γGlu-CO(CH ₂ ) ₁₆ CO ₂ H; and two cysteine residues, designated C*, at positions 2 and 7 are additionally linked by a disulfide bridge. 11. The isolated polypeptide according to claim 5, comprising the amino acid sequence: KC*NTSTC*ATQRLANELHKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 130), or a pharmaceutically acceptable salt thereof, where two cysteine residues, designated C*, at positions 2 and 7 are additionally linked by a disulfide bridge. 12. The isolated polypeptide according to claim 5, comprising the amino acid sequence: KC*NTSTC*ATQRLANFLQKSSNNFGPILPPTKVGSETY-(NH ₂ ) (SEQ ID NO: 131), or a pharmaceutically acceptable salt thereof, where two cysteine residues, designated C*, at positions 2 and 7 are additionally linked by a disulfide bridge. 13. An isolated polypeptide according to any one of claims 1-4 or a pharmaceutically acceptable salt thereof, comprising a lipophilic substituent and optionally comprising a spacer. 14. The isolated polypeptide according to claim 13, or a pharmaceutically acceptable salt thereof, containing a lipophilic substituent and a spacer of formula VI: -(Y1) _n1 -(V) _r -(Y2) _n2 -CO-(CH ₂ ) _m -Z Formula VI, or a lipophilic substituent and a spacer of formula III: -(γGlu) _n -CO-(CH ₂ ) _m -Z ((γGlu) _n " designated as SEQ ID NO: 311) Formula III, Where Z represents _-CH3 or _-CO2H ; m is from 4 to 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 (O( _CH2 ) ₂ ) _tOCH2NH ]-, and t is from 1 to ₈ ; r is from 1 to 8; n is from 1 to 10; n1 is from 0 to 10; and n2 is from 0 to 10. 15. A pharmaceutical composition for treating a metabolic disorder selected from the group consisting of type 1 diabetes, type 2 diabetes and obesity in a human subject, comprising an effective amount of the isolated polypeptide according to any one of claims 1-14 or a pharmaceutically acceptable salt thereof. 16. An osmotic delivery device for subcutaneously or subdermally administering a pharmaceutical composition according to claim 15, wherein said osmotic delivery device comprises: an impermeable reservoir comprising inner and outer surfaces and first and second open ends; a semipermeable membrane hermetically attached to the first open end of the reservoir; an osmotic engine located inside the reservoir and adjacent to a semipermeable membrane; a piston adjacent to the osmotic engine, wherein the piston forms a movable seal with the inner surface of the reservoir, the piston divides the reservoir into a first chamber and a second chamber, wherein the first chamber contains the osmotic engine; a medicinal composition or a composition in the form of a suspension containing an isolated polypeptide or a pharmaceutical composition, wherein the second chamber contains a medicinal product or a composition in the form of a suspension, and the medicinal product or composition in the form of a suspension is in liquid form; and a diffusion retarder inserted into the second open end of the reservoir, the diffusion retarder being adjacent to the composition in the form of a suspension. 17. A method for treating obesity in a human subject, comprising administering to said subject an isolated polypeptide according to any one of claims 1-14 or a pharmaceutically acceptable salt thereof, a pharmaceutical composition according to claim 15, or an osmotic device according to claim 16. 18. A method for treating type 1 or type 2 diabetes in a human subject, comprising administering to the subject an isolated polypeptide according to any one of claims 1-14 or a pharmaceutically acceptable salt thereof, a pharmaceutical composition according to claim 15, or an osmotic device according to claim 16. 19. A pharmaceutical composition for providing weight loss or appetite suppression in a human subject, comprising an effective amount of the isolated polypeptide according to any one of claims 1-14 or a pharmaceutically acceptable salt thereof. 20. A method for providing weight loss or appetite suppression in a human subject, comprising administering to the subject an isolated polypeptide according to any one of claims 1-14, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 19.

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