# US 10,501,517 B2 · Glucagon-receptor selective polypeptides and methods of use thereof

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- Number: US 10,501,517 B2
- Document ID: US10501517B2
- Status: Granted
- Kind: B2
- Application number: 15/595,809
- Filed: 2017-05-15
- Published: 2018-01-11
- Granted: 2019-12-10
- Earliest claimed: 2016-05-16
- Assignees: Intarcia Therapeutics, Inc.
- Applicants: Intarcia Therapeutics Inc.
- Inventors: William Blackwell; Ved P. Srivastava; Mark A. Paulik; Andrew Young; Robert Neil Hunter, III; Steven Thomas Dock
- Web page: https://patentgrep.com/patent/US10501517B2

## CPC

- C07K 14/605: Glucagons
- A61K 9/0004: {Osmotic delivery systems; Sustained release driven by osmosis, thermal energy or gas}
- A61K 38/26: Glucagons
- A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
- A61P 3/00: Drugs for disorders of the metabolism (of the blood or the extracellular fluid A61P7/00)
- A61P 3/04: Anorexiants; Antiobesity agents
- A61P 3/08: for glucose homeostasis (pancreatic hormones A61P5/48)
- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- A61P 5/00: Drugs for disorders of the endocrine system
- A61P 7/00: Drugs for disorders of the blood or the extracellular fluid
- A61P 7/04: Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
- A61P 9/00: Drugs for disorders of the cardiovascular system
- A61P 11/00: Drugs for disorders of the respiratory system
- A61P 13/12: of the kidneys
- A61P 25/00: Drugs for disorders of the nervous system
- A61P 25/04: Centrally acting analgesics, e.g. opioids
- A61P 25/28: for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P 35/00: Antineoplastic agents
- A61P 37/02: Immunomodulators
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C07K 14/00: Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61M 5/14276: {specially adapted for implantation}
- A61M 31/002: {Devices for releasing a drug at a continuous and controlled rate for a prolonged period of time (artificial gland structures or devices A61F2/022; intra-uterine contraceptive devices A61F6/14; tampons for introducing into the vagina A61F13/20, A61L15/00; suppositories or bougies for intra-vaginal or intra-uterine application A61K9/02; physical forms of medicinal preparations for sustained or differential drug release A61K9/20, A61K9/50)}
- A61M 2005/14513: {with secondary fluid driving or regulating the infusion}
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61M: DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY (introducing media into or onto the bodies of animals A61D7/00; means for inserting tampons A61F13/26; devices for administering food or medicines orally A61J; containers for collecting, storing or administering blood or medical fluids A61J1/05); DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY (surgery A61B; chemical aspects of surgical articles A61L); DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR {(Electrotherapy, e.g. producing anaesthesia by the use of alternating or intermittent currents A61N1/36021)}

## Abstract

This invention relates to isolated polypeptides that are glucagon-receptor selective analogs and peptide derivatives thereof. These analogs are selective for human glucagon receptor with improved solubility, thermal stability, and physicochemical properties as compared to native endogenous glucagon. This invention also relates to methods of using such polypeptides in a variety of therapeutic and diagnostic indications, as well as methods of producing such polypeptides. These analogs are useful, alone or in combination with other therapeutic peptides, in methods of treating obesity, diabetes, metabolic disorders, and other disorders or disease.

## Claims

### Claim 1

An isolated polypeptide comprising:
(i) the amino acid sequence:
X1X2X3GTFTSDX10X11X12X13LX15X16X17X18AQEFX23X24X25LEDE-Z-tail (SEQ ID NO: 1), wherein:
X1 is Y or W;
X2 is S, G or T;
X3 is Q or H;
X10 is Y or H;
X11 is S or T;
X12 is K or R;
X13 is Y, L or W;
X15 is D or E;
X16 is S, 2-Aminoisobutyric acid (Aib), A, E, L, Q, K, or I;
X17 is K, E, S, T, or A;
X18 is A, R, S, E, L, T or Y;
X23 is T or V;
X24 is K, I, L, or Aib;
X25 is H or W; and
Z-tail is absent; or is selected from the group consisting of EEPSSGAPPPS-OH (SEQ ID NO: 4); EPSSGAPPPS-OH (SEQ ID NO: 5); GAPPPS-OH (SEQ ID NO: 6); GGPSSGAPPPS-OH (SEQ ID NO: 7); GPSSGAPPPS-OH (SEQ ID NO: 8); KRNKNPPPS-OH (SEQ ID NO: 9); KRNKNPPS-OH (SEQ ID NO: 10); KRNKPPIA-OH (SEQ ID NO: 11); KRNKPPPA-OH (SEQ ID NO: 150); KRNKPPPS-OH (SEQ ID NO: 12); KSSGKPPPS-OH (SEQ ID NO: 13); PESGAPPPS-OH (SEQ ID NO: 14); PKSGAPPPS-OH (SEQ ID NO: 15); PKSKAPPPS-NH2 (SEQ ID NO: 16); PKSKAPPPS-OH (SEQ ID NO: 17); PKSKEPPPS-NH2 (SEQ ID NO: 18); PKSKEPPPS-OH (SEQ ID NO: 19); PKSKQPPPS-OH (SEQ ID NO: 20); PKSKSPPPS-NH2 (SEQ ID NO: 21); PKSKSPPPS-OH (SEQ ID NO: 22); PRNKNNPPS-OH (SEQ ID NO: 23); PSKGAPPPS-OH (SEQ ID NO: 24); PSSGAPPPSE-OH (SEQ ID NO: 25); PSSGAPPPS-NH2 (SEQ ID NO: 26); PSSGAPPPS-OH (SEQ ID NO: 27); PSSGAPPPSS-OH (SEQ ID NO: 28); PSSGEPPPS-OH (SEQ ID NO: 29); PSSGKKPPS-OH (SEQ ID NO: 30); PSSGKPPPS-NH2 (SEQ ID NO: 31); PSSGKPPPS-OH (SEQ ID NO: 32); PSSGSPPPS-OH (SEQ ID NO: 33); PSSKAPPPS-OH (SEQ ID NO: 34); PSSKEPPPS-OH (SEQ ID NO: 35); PSSKGAPPPS-OH (SEQ ID NO: 36); PSSKQPPPS-OH (SEQ ID NO: 37); PSSKSPPPS-OH (SEQ ID NO: 38); SGAPPPS-OH (SEQ ID NO: 39); and SSGAPPPS-OH (SEQ ID NO: 40); or
(ii) the amino acid sequence:
YSQGTFTSDYSKYLDSX17RAQX21FVX24WLX27X28T-OH (SEQ ID NO: 137),
wherein:
X17 is K*, where K* is in a lactam bridge with E* at X21;
X21 is E*, where E* is in a lactam bridge with K* at X17;
X24 is K or K**, where K** is in a lactam bridge with E** at X28;
X27 is Q or D; and
X28 is E or E**, where E** is in a lactam bridge with K** at X24; or
(iii) the amino acid sequence:
X1SX3GTFTSDX10SKYLDX16X17X18AQX21FVX24WLEDEPX31SX33X34PPPS-OH (SEQ ID NO: 151),
wherein:
X1=Y or W;
X3=H or Q;
X10=Y, K or K***;
X16=A, S, Aib, K or K***;
X17=A, K, Aib or K***;
X18=Y, S or R;
X21=E, K or K***;
X24=I, Aib, K or K***;
X31=S, K or K***;
X33=G, K or K***; and
X34=A or S;
wherein K*** is lysine, and the ε-amino group of the lysine sidechain is covalently joined to a lipophilic substituent, optionally via a spacer.

### Claim 2 (depends on 1)

The isolated polypeptide of claim 1, comprising the amino acid sequence:
X1X2X3GTFTSDX10X11X12X13LX15X16X17X18AQEFX23X24X25LEDE-Z-tail (SEQ ID NO: 1), wherein:
X1 is Y or W;
X2 is S, G or T;
X3 is Q or H;
X10 is Y or H;
X11 is S or T;
X12 is K or R;
X13 is Y, L or W;
X15 is D or E;
X16 is S, 2-Aminoisobutyric acid (Aib), A, E, L, Q, K, or I;
X17 is K, E, S, T, or A;
X18 is A, R, S, E, L, T or Y;
X23 is T or V;
X24 is K, I, L, or Aib;
X25 is H or W; and
Z-tail is absent; or is selected from the group consisting of EEPSSGAPPPS-OH (SEQ ID NO: 4); EPSSGAPPPS-OH (SEQ ID NO: 5); GAPPPS-OH (SEQ ID NO: 6); GGPSSGAPPPS-OH (SEQ ID NO: 7); GPSSGAPPPS-OH (SEQ ID NO: 8); KRNKNPPPS-OH (SEQ ID NO: 9); KRNKNPPS-OH (SEQ ID NO: 10); KRNKPPIA-OH (SEQ ID NO: 11); KRNKPPPA-OH (SEQ ID NO: 150); KRNKPPPS-OH (SEQ ID NO: 12); KSSGKPPPS-OH (SEQ ID NO: 13); PESGAPPPS-OH (SEQ ID NO: 14); PKSGAPPPS-OH (SEQ ID NO: 15); PKSKAPPPS-NH2 (SEQ ID NO: 16); PKSKAPPPS-OH (SEQ ID NO: 17); PKSKEPPPS-NH2 (SEQ ID NO: 18); PKSKEPPPS-OH (SEQ ID NO: 19); PKSKQPPPS-OH (SEQ ID NO: 20); PKSKSPPPS-NH2 (SEQ ID NO: 21); PKSKSPPPS-OH (SEQ ID NO: 22); PRNKNNPPS-OH (SEQ ID NO: 23); PSKGAPPPS-OH (SEQ ID NO: 24); PSSGAPPPSE-OH (SEQ ID NO: 25); PSSGAPPPS-NH2 (SEQ ID NO: 26); PSSGAPPPS-OH (SEQ ID NO: 27); PSSGAPPPSS-OH (SEQ ID NO: 28); PSSGEPPPS-OH (SEQ ID NO: 29); PSSGKKPPS-OH (SEQ ID NO: 30); PSSGKPPPS-NH2 (SEQ ID NO: 31); PSSGKPPPS-OH (SEQ ID NO: 32); PSSGSPPPS-OH (SEQ ID NO: 33); PSSKAPPPS-OH (SEQ ID NO: 34); PSSKEPPPS-OH (SEQ ID NO: 35); PSSKGAPPPS-OH (SEQ ID NO: 36); PSSKQPPPS-OH (SEQ ID NO: 37); PSSKSPPPS-OH (SEQ ID NO: 38); SGAPPPS-OH (SEQ ID NO: 39); and SSGAPPPS-OH(SEQ ID NO: 40).

### Claim 3 (depends on 1)

The isolated polypeptide of claim 1, comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 41-48 50-136 and 143-149 (Compounds A1-A8 and A10-A103).

### Claim 4 (depends on 1)

The isolated polypeptide of claim 1, comprising the amino acid sequence:
X1X2X3GTFTSDX10X11X12X13LX15X16X17X18AQEFVX24WLEDE-Z-tail (SEQ ID NO: 2), wherein:
X1 is Y or W;
X2 is S or G;
X3 is Q or H;
X10 is Y or H;
X11 is S or T;
X12 is K or R;
X13 is Y, L or W;
X15 is D or E;
X16 is 2-Aminoisobutyric acid (Aib), A, or S;
X17 is A or K;
X18 is R, S, L, or Y;
X24 is K, I, or Aib; and
Z-tail is absent or is selected from the group consisting of PSSGAPPPS-NH2 (SEQ ID NO: 26); PSSGAPPPS-OH (SEQ ID NO: 27); and PKSKSPPPS-NH2 (SEQ ID NO: 21).

### Claim 5 (depends on 4)

The isolated polypeptide of claim 4, comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 41-43 and 143-149 (Compounds A1-A3 and A97-103).

### Claim 6 (depends on 1)

The isolated polypeptide of claim 1, comprising the amino acid sequence:
YSX3GTFTSDYSKYLDX16X17X18AQEFVX24WLEDE-Z-tail (SEQ ID NO: 3), wherein:
X3 is Q or H;
X16 is 2-Aminoisobutyric acid (Aib) or A;
X17 is A or K;
X18 is R, S, or Y;
X24 is K or Aib; and
Z-tail is selected from the group consisting of PSSGAPPPS-OH (SEQ ID NO: 27) and PKSKSPPPS-NH2 (SEQ ID NO: 21).

### Claim 7 (depends on 6)

The isolated polypeptide of claim 6, comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 41-43 (Compounds A1-A3).

### Claim 8 (depends on 6)

The isolated polypeptide of claim 6, comprising an amino acid sequence of SEQ ID NO: 41 (Compound A1).

### Claim 9 (depends on 6)

The isolated polypeptide of claim 6, comprising an amino acid sequence of SEQ ID NO: 42 (Compound A2).

### Claim 10 (depends on 6)

The isolated polypeptide of claim 6, comprising an amino acid sequence of SEQ ID NO: 43 (Compound A3).

### Claim 11 (depends on 6)

The isolated polypeptide of claim 6, consisting of an amino acid sequence of SEQ ID NO: 41 (Compound A1).

### Claim 12 (depends on 6)

The isolated polypeptide of claim 6, consisting of an amino acid sequence of SEQ ID NO: 42 (Compound A2).

### Claim 13 (depends on 6)

The isolated polypeptide of claim 6, consisting of an amino acid sequence of SEQ ID NO: 43 (Compound A3).

### Claim 14 (depends on 1)

The isolated polypeptide of claim 1, comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 44-46 (Compounds A4-A6).

### Claim 15 (depends on 1)

The isolated polypeptide of claim 1, comprising an amino acid sequence of SEQ ID NO: 44 (Compound A4).

### Claim 16 (depends on 1)

The isolated polypeptide of claim 1, comprising an amino acid sequence of SEQ ID NO: 45 (Compound A5).

### Claim 17 (depends on 1)

The isolated polypeptide of claim 1, comprising an amino acid sequence of SEQ ID NO: 46 (Compound A6).

### Claim 18 (depends on 1)

The isolated polypeptide of claim 1, comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 47, 48, and 50-136 (Compounds A7, A8, and A10-A96).

### Claim 19 (depends on 1)

The isolated polypeptide of claim 1, comprising the amino acid sequence:
YSQGTFTSDYSKYLDSX17RAQX21FVX24WLX27X28T-OH (SEQ ID NO: 137), wherein:
X17 is K*, where K* is in a lactam bridge with E* at X21;
X21 is E*, where E* is in a lactam bridge with K* at X17;
X24 is K or K**, where K** is in a lactam bridge with E** at X28;
X27 is Q or D; and
X28 is E or E**, where E** is in a lactam bridge with K** at X24.

### Claim 20 (depends on 19)

The isolated polypeptide of claim 19, comprising the amino acid sequence of SEQ ID NO: 138 (Compound A104) or SEQ ID NO: 139 (Compound A105).

### Claim 21 (depends on 1)

The isolated polypeptide of claim 1, comprising the amino acid sequence:
X1SX3GTFTSDX10SKYLDX16X17X18AQX21FVX24WLEDEPX31SX33X34PPPS-OH (SEQ ID NO: 151),
wherein: X1=Y or W;
X3=H or Q;
X10=Y, K or K***;
X16=A, S, Aib, K or K***;
X17=A, K, Aib or K***;
X18=Y, S or R;
X21=E, K or K***;
X24=I, Aib, K or K***;
X31=S, K or K***;
X33=G, K or K***; and
X34=A or S;
wherein K*** is lysine, and the ε-amino group of the lysine sidechain is covalently joined to a lipophilic substituent, optionally via a spacer.

### Claim 22 (depends on 21)

The isolated polypeptide of claim 21, wherein each lipophilic substituent is covalently bound to the peptide via a spacer, and wherein each lipophilic substituent and spacer are of Formula II:

### Claim 23 (depends on 21)

The isolated polypeptide of claim 21, wherein each lipophilic substituent is covalently bound to the peptide via a spacer, and wherein each lipophilic substituent and spacer are of Formula III:

### Claim 24 (depends on 21)

The isolated polypeptide of claim 21, selected from the group consisting of SEQ ID NOs: 152-183 (Compounds B1-B32).

### Claim 25 (depends on 24)

The isolated polypeptide of claim 24, selected from the group consisting of SEQ ID NOs: 154 (Compound B3), 157 (Compound B6), 158 (Compound B7), 160 (Compound B9), 161 (Compound B10), 162 (Compound B11), 165 (Compound B14), 166 (Compound B15), 168 (Compound B17), 172 (Compound B21), and 173 (Compound B22).

### Claim 26 (depends on 21)

The isolated polypeptide of claim 21, wherein the isolated peptide is SEQ ID NO: 160 (Compound B9), 165 (Compound B14) or 166 (Compound B15).

### Claim 27 (depends on 1)

A pharmaceutical composition comprising the isolated polypeptide of claim 1 or a pharmaceutically acceptable salt thereof.

### Claim 28 (depends on 27)

The pharmaceutical composition of claim 27, further comprising a second polypeptide.

### Claim 29 (depends on 28)

The pharmaceutical composition of claim 28, wherein the second polypeptide is an insulinotropic polypeptide.

### Claim 30 (depends on 28)

The pharmaceutical composition of claim 28, wherein the second polypeptide is selected from the group consisting of exenatide, a derivative of exenatide, an analog of exenatide, glucagon-like peptide-1 (GLP-1), a derivative of GLP-1, and an analog of GLP-1.

### Claim 31 (depends on 28)

The pharmaceutical composition of claim 28, wherein the second polypeptide is exenatide.

### Claim 32 (depends on 28)

The pharmaceutical composition of claim 28, wherein the second polypeptide is a GLP-1 analog.

### Claim 33 (depends on 27)

The pharmaceutical composition of claim 27, wherein the pharmaceutically acceptable salt is a trifluoroacetate salt, acetate salt or hydrochloride salt.

### Claim 34 (depends on 27)

The pharmaceutical composition of claim 27, wherein the isolated polypeptide is selected from the group consisting of SEQ ID NOs: 41-43 (Compounds A1-A3).

### Claim 35 (depends on 27)

The pharmaceutical composition of claim 27, wherein the isolated polypeptide comprises an amino acid sequence of SEQ ID NO: 41 (Compound A1), and wherein the pharmaceutically acceptable salt is a trifluoroacetate salt, acetate salt, or hydrochloride salt.

### Claim 36 (depends on 1)

A particle formulation comprising the isolated polypeptide of claim 1 and at least one stabilizing component.

### Claim 37 (depends on 36)

The particle formulation of claim 36, wherein the at least one stabilizing component is selected from the group consisting of one or more carbohydrates, one or more antioxidants, one or more amino acids, one or more buffers, one or more inorganic compounds, one or more surfactants, and combinations thereof.

### Claim 38 (depends on 36)

A suspension formulation comprising the particle formulation of claim 36 and a suspension vehicle.

### Claim 39 (depends on 1)

An osmotic delivery device, comprising the isolated polypeptide of claim 1.

### Claim 40 (depends on 39)

The osmotic delivery device of claim 39, further comprising
an impermeable reservoir comprising interior and exterior surfaces and first and second open ends,
a semi-permeable membrane in sealing relationship with the first open end of the reservoir,
an osmotic engine within the reservoir and adjacent the semi-permeable membrane,
a piston adjacent the osmotic engine, wherein the piston forms a movable seal with the interior surface of the reservoir, the piston divides the reservoir into a first chamber and a second chamber, the first chamber comprising the osmotic engine,
a suspension formulation, wherein the second chamber comprises the suspension formulation and the suspension formulation is flowable and comprises the isolated polypeptide, and
a diffusion moderator inserted in the second open end of the reservoir, the diffusion moderator adjacent the suspension formulation.

### Claim 41 (depends on 40)

The osmotic delivery device of claim 40, wherein the reservoir comprises titanium or a titanium alloy.

### Claim 42 (depends on 36, 40)

The osmotic delivery device of claim 40, wherein the suspension formulation comprises the particle formulation of claim 36 and a suspension vehicle.

### Claim 43 (depends on 1)

A pharmaceutical composition comprising the isolated polypeptide of claim 1, wherein the isolated polypeptide is a glucagon analog and the composition further comprises an additional agent selected from the group consisting of exenatide, a derivative of exenatide, an analogue of exenatide, glucagon-like peptide-1 (GLP-1), a derivative of GLP-1, and an analogue of GLP-1.

### Claim 44 (depends on 43)

The pharmaceutical composition of claim 43, wherein the additional agent is exenatide.

### Claim 45 (depends on 43)

The pharmaceutical composition of claim 43, wherein the glucagon analog is selected from the group consisting of SEQ ID NOs: 41-48, 50-136, 138, 139, and 143-149 (Compounds A1-A8 and A10-A105).

### Claim 46 (depends on 45)

The pharmaceutical composition of claim 45, wherein the glucagon analog is selected from the group consisting of SEQ ID NOs: 41, 42, 43, 44, 45, and 46 (Compounds A1, A2, A3, A4, A5 and A6).

### Claim 47 (depends on 43)

The pharmaceutical composition of claim 43, wherein the glucagon analog is selected from the group consisting of SEQ ID NOs: 152-183 (Compounds B1-B32).

### Claim 48 (depends on 47)

The pharmaceutical composition of claim 47, wherein the glucagon analog is selected from the group consisting of SEQ ID NOs: 154 (Compound B3), 157 (Compound B6), 158 (Compound B7), 160 (Compound B9), 161 (Compound B10), 162 (Compound B11), 165 (Compound B14), 166 (Compound B15), 168 (Compound B17), 172 (Compound B21), and 173 (Compound B22).

### Claim 49 (depends on 43)

The pharmaceutical composition of claim 43, wherein the glucagon analog is formulated in combination with exenatide in a fixed ratio of glucagon analog:exenatide of 1000:1 to 1:1000.

### Claim 50 (depends on 43)

The pharmaceutical composition of claim 43, wherein the glucagon analog is formulated in combination with exenatide in a fixed ratio of glucagon analog:exenatide of 100:1 to 1:100.

### Claim 51 (depends on 43)

The pharmaceutical composition of claim 43, wherein the glucagon analog is formulated in combination with exenatide in a fixed ratio of glucagon analog:exenatide of 10:1 to 1:10.

### Claim 52

An isolated polypeptide, wherein the isolated polypeptide is selected from the group consisting of SEQ ID NOs: 41-136, 138, 139, and 143-149 (Compounds A1-A105), and the isolated polypeptide is covalently joined to a lipophilic substituent, optionally via a spacer.

### Claim 53 (depends on 52)

The isolated polypeptide of claim 52, wherein the isolated polypeptide is selected from the group consisting of SEQ ID NOs: 41-46 (Compounds A1-A6).

### Claim 54 (depends on 52)

The isolated polypeptide of claim 52, wherein the lipophilic substituent is covalently bound to the peptide via a spacer, and the lipophilic substituent and spacer are of Formula II:

### Claim 55 (depends on 52)

The isolated polypeptide of claim 52, wherein the lipophilic substituent is covalently bound to the peptide via a spacer, and the lipophilic substituent and spacer are of Formula III:

### Claim 56 (depends on 1)

A method of treating or alleviating a symptom of a disease or disorder associated with aberrant glucagon activity in a subject in need thereof, the method comprising administering to the subject the isolated polypeptide of claim 1 or a pharmaceutical composition comprising the isolated polypeptide thereof.

### Claim 57 (depends on 56)

The method of claim 56, wherein the disease or disorder is type 2 diabetes mellitus.

### Claim 58 (depends on 56)

The method of claim 56, wherein the subject is a human.

### Claim 59 (depends on 56)

The method of claim 56, wherein the isolated polypeptide or pharmaceutical composition is formulated for administration by a route selected from the group consisting of parenteral, oral, transdermal, transmucosal, rectal, intravenous, intradermal, subdermal, subcutaneous, and any combination thereof.

### Claim 60 (depends on 1)

A method of:
(i) treating a metabolic disorder, obesity, metabolic disease or disorder associated with elevated blood glucose or
(ii) treating, delaying the onset of, delaying the progression of, or otherwise ameliorating a symptom of a disease or disorder in which co-agonism at both the glucagon and GLP-1 receptors is desired,
said method comprising administering the isolated polypeptide of claim 1 to a subject in need thereof.

### Claim 61 (depends on 60)

The method of claim 60, wherein the metabolic disorder is type 2 diabetes mellitus.