GLP-1 analogs useful for diabetes treatment

• US 5,545,618 A
• Filed: 12/10/1993
• Issued: 08/13/1996
• Est. Priority Date: 01/24/1990
• Status: Expired due to Term

• Current Assignee: Scios, Inc. (Johnson & Johnson), The General Hospital Corporation (Mass General Brigham, Inc.)
• Original Assignee: Douglas I. Buckley, Joanne B. Mallory, Joel F. Habener, Svetlana Mojsov
• Inventors: Habener, Joel F., Mojsov, Svetlana, Buckley, Douglas I., Mallory, Joanne B.
• Primary Examiner(s): Nucker, Christine M.
• Assistant Examiner(s): Prickril, Benet
• Application Number:
  US08/165,516
• Time in Patent Office: 977 Days
• Field of Search: 514/12, 530/324
• US Class Current: 514/5.9
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 14/605 Glucagons

Abstract

The invention provides effective analogs of the active GLP-1 peptides, 7-34, 7-35, 7-36, and 7-37, which have improved characteristics for treatment of diabetes Type II. These analogs have amino acid substitutions at positions 7-10 and/or are truncated at the C-terminus and/or contain various other amino acid substitutions in the basic peptide. The analogs may either have an enhanced capacity to stimulate insulin production as compared to glucagon or may exhibit enhanced stability in plasma as compared to GLP-1 (7-37) or both. Either of these properties will enhance the potency of the analog as a therapeutic. Analogs having D-amino acid substitutions in the 7 and 8 positions and/or N-alkylated or N-acylated amino acids in the 7 position are particularly resistant to degradation in vivo.

Claims

1. A peptide which is more potent than glucagon in stimulating insulin release from islet cells, said peptide consisting of labeled or unlabeled GLP-1(7-34), GLP-1(7-35), GLP-1(7-36), or GLP-1(7-37) or the C-terminal amide form thereof, having at least one modification selected from the group consisting of:

(a) substitution of a neutral amino acid, arginine, or a D form of lysine for lysine at position 26 and/or 34 and/or a neutral amino acid, lysine, or a D form of arginine for arginine at position 36;

(b) substitution of an oxidation-resistant amino acid for tryptophan at position

(c) substitution according to at least one of;

Y for V at position 16;

K for S at position 18;

D for E at position 21;

S for G at position 22;

R for Q at position 23;

R for A at position 24; and

Q for K at position 26;

(d) a substitution consisting of at least one of;

an alternative small neutral amino acid for A at position 8;

an alternative acidic amino acid or neutral amino acid for E at position 9;

an alternative neutral amino acid for G at position 10; and

an alternative acidic amino acid for D at position 15; and

(c) substitution of an alternative neutral amino acid or the D or N-acylated or alkylated form of histidine for histidine at position 7

wherein for (a), (b), (d) and (e), the substituted amino acids can optionally be in the D form and the amino acids substituted at position 7 can optionally be in the N-acylated or N-alkylated form.

2. The peptide of claim 1 wherein the only modification is as set forth in paragraph (a) of claim 1 and wherein the amino acid substituted for lysine at positions 26 and/or 34 is selected from the group consisting of K.sup.†

, G, S, A, L, I, Q, M, R and R.sup.†

and the amino acid substituted for arginine at position 36 is selected from the group consisting of K, K.sup.†

G, S, A, L, I, Q, M, and R.sup.†

,

optionally in combination with a modification as set forth in paragraph (b), (c), (d) or (e) of claim 1.

3. The peptide of claim 1 wherein the only modification is as set forth in paragraph (b) of claim 1 and wherein the amino acid substituted for tryptophan at position 31 is selected from the group consisting of F, V, L, I, A and Y,

optionally in combination with a modification as set forth in paragraph (a), (c), (d) or (e) of claim 1.

4. The peptide of claim 1 wherein the only modification is as set forth in paragraph (c) of claim 1 and wherein combined substitutions of S for G at position 22, K at positions 23 and 24 for Q and A, respectively, and Q for K at position 26 have been made, or substitutions of Y for V at position 16 and K for S at position 18 have been made, or these substitutions plus D for E at positions 21 have been made,

optionally in combination with a modification as set forth in paragraph (a), (b), (d) or (e) of claim 1.

5. The peptide of claim 1 wherein the only modification is as set forth in paragraph (d) of claim 1 and wherein the small neutral amino acid substituted for alanine at position 8 is selected from the group consisting of S, S.sup.†

, G, C, C.sup.†

, Sar, A.sup..sup.†

, beta-ala, and Aib and the acidic or neutral amino acid substituted for glutamic acid at position 9 is selected from the group consisting of E.sup.†

, D, D.sup.†

, Cya, T, T.sup.†

, N, N.sup.†

, Q, Q.sup.†

, Cit, MSO, and acetyl-K, and the alternative neutral amino acid substituted for glycine at position 10 is selected from the group consisting of S, S.sup.†

, Y, Y.sup.†

, T, T.sup.†

, N, N.sup.†

, Q, Q.sup.†

, Git, MSO, acetyl-K, F, and F.sup.†

,

optionally in combination with a modification as set forth in paragraph (a), (b), (c) or (e) of claim 1.

6. The peptide of claim 1 wherein the only modification is as set forth in paragraph (e) of claim 1 and wherein the amino acid substituted for histidine at position 7 is selected from the group consisting of H.sup.†

, Y, Y.sup.†

, F, F.sup.†

, R, R.sup.†

, Orn, Orn.sup.†

, M, M.sup.†

, N-formyl-H, N-formyl-H.sup.†

, N-acetyl-H, N-acetyl-H.sup.†

, N-isopropyl-H, N-isopropyl-H.sup.†

, N-acetyl-K;

N-acetyl-K.sup.†

, P, and P.sup.†

,

optionally in combination with a modification as set forth in paragraph (a), (b), (c), or (d) of claim 1.

7. The peptide of claim 1 which is selected from the group consisting of

(H.sup.†

)⁷ -GLP-1(7-37),

(Y)⁷ -GLP-1(7-37),

(N-acetyl-H)⁷ -GLP-1(7-37),

(N-isopropyl-H)⁷ -GLP-1(7-37),

(A.sup.†

)⁸ -GLP-1(7-37),

(E.sup.†

)⁹ -GLP-1(7-37),

(D)⁹ -GLP-1(7-37),

(D.sup.†

)⁹ -GLP-1(7-37),

(F.sup.†

)¹⁰ -GLP-1(7-37),

(S)²² (R)²³ (R)²⁴ (Q)²⁶ -GLP-1 (7-37), and

(S)⁸ (Q)⁹ (Y)¹⁶ (K)¹⁸ (D)²¹ -GLP-1(7-37).

12. A pharmaceutical composition useful in the treatment of diabetes Type II which comprises an effective amount of the peptide of claim 1 or 8 in admixture with a pharmaceutically acceptable excipient.

8. A peptide useful to release insulin from islet cells, said peptide having enhanced resistance to degradation in plasma as compared to GLP-1(7-37) and said peptide consisting of labeled or unlabeled GLP-1(7-34), GLP-1(7-35), GLP-I(7-36), or GLP-1(7-37) or the C-terminal amide form thereof, having at least one modification selected from the group consisting of:

(a) substitution of the D form of a neutral or acidic amino acid or the D form of histidine for histidine at position 7;

(b) substitution of a D form of an amino acid for alanine at position 8; and

(c) substitution of an N-acylated (1-6C) or N-alkylated (1-6C) form of an alternate amino acid or of histidine for histidine at position 7.

9. The peptide of claim 8 wherein the only modification is as set forth in paragraph (a) of claim 8 and wherein the D form of the amino acid substituted for histidine at position 7 is selected from the group consisting of P.sup.†

, D.sup.†

, E.sup.†

, N.sup.†

, Q.sup.†

, L.sup.†

, V.sup.†

, I.sup.†

, and H.sup.†

,

optionally in combination with a modification as set forth in paragraph (b) or (c) of claim 8.

10. The peptide of claim 8 wherein the only modification is as set forth in paragraph (b) of claim 8 and wherein the D-amino acid at position 8 is selected from the group consisting of P.sup.†

, V.sup.†

, L.sup.†

, I.sup.†

, and A.sup.†

,

optionally in combination with a modification as set forth in paragraph of claim 8.

11. The peptide of claim 8 wherein the only modification is as set forth in paragraph (c) of claim 8 and wherein the alkylated or acetylated amino acid is selected from the group consisting of P, D, E, N, Q, V, L, I, K and H,

optionally in combination with a modification as set forth in paragraph (a) or (b) of claim 8.

13. The peptide of claim 8 which is selected from the group consisting of

(H.sup.†

)⁷ -GLP-1(7-37),

(N-acetyl-H)⁷ -GLP-1(7-37),

(N-isopropyl-H)⁷ -GLP-1(7-37),

(N-acetyl-K)⁷ -GLP-1(7-37), and

(A.sup.†

)⁸ -GLP-1(7-37).

14. A peptide which is more potent than glucagon in stimulating insulin release from islet cells, and said peptide consisting of labeled or unlabeled analogs of truncated forms of GLP-1, which consist of GLP-1(7-34), GLP-1(7-35), GLP-1(7-36), or GLP-1(7-37) or the C-terminal amide form thereof, wherein the analog is selected from the group consisting of:

(H.sup.†

)⁷ -GLP-1(7-37)

(Y)⁷ GLP-1(7-37),

(N-acetyl-H)⁷ -GLP-1(7-37),

(N-isopropyl-H)⁷ -GLP-1(7-37),

(A.sup.†

)⁸ -GLP-1(7-37),

(E.sup.†

)⁹ -GLP-1(7-37),

(D)⁹ -GLP-1(7-37),

(D.sup.†

)⁹ -GLP-1(7-37),

(F.sup.†

)¹⁰ -GLP-1(7-37),

(S)²² (R)²³ (R)²⁴ (Q)²⁶ -GLP-1(7-37), and

(S)⁸ (Q)⁹ (Y)¹⁶ (K)¹⁸ (D)²¹ -GLP-1(7-37).