Insulin derivatives having an extremely delayed time-action profile
The invention relates to novel insulin analogs having a basal time-action profile, which are characterized by the following features: a) the B chain end consists of an amidated basic amino acid residue such as lysine or arginine amide; b) the N-terminal amino acid residue of the insulin A chain is a lysine or arginine radical; c) the amino acid position A8 is occupied by a histidine radical; d) the amino acid position A21 is occupied by a glycine radical; and e) one or more substitutions and/or additions of negatively charged amino acid residues are carried out in the positions A5, A15, A18, B-1, B0, B1, B2, B3 and B4.
1. An insulin analogue of the formula:
wherein
A0 corresponds to Lys or Arg;
A5 corresponds to Asp, Gln or Glu;
A15 corresponds to Asp, Glu or Gln;
A18 corresponds to Asp, Glu or Asn;
B-1 corresponds to Asp, Glu;
B0 corresponds to Asp, Glu or a chemical bond;
B1 corresponds to Asp, Glu or Phe;
B2 corresponds to Asp, Glu or Val;
B3 corresponds to Asp, Glu or Asn;
B4 corresponds to Asp, Glu or Gln;
B29 corresponds to Lys or a chemical bond;
B30 corresponds to Thr or a chemical bond;
B31 corresponds to Arg, Lys or a chemical bond;
B32 corresponds to Arg-amide or Lys-amide; and
wherein two amino acid residues of the group comprising A5, A15, A18, B-1, B0, B1, B2, B3 and B4 correspond simultaneously and independently of one another to Asp or Glu.
2. The insulin analogue as claimed in claim 1 , wherein A0 corresponds to Arg.
3. The insulin analogue as claimed in claim 1 , wherein A5 corresponds to Glu.
4. The insulin analogue as claimed in claim 1 , wherein A15 corresponds to Glu.
5. The insulin analogue as claimed in claim 1 , wherein A18 corresponds to Asp.
6. The insulin analogue as claimed in claim 1 , wherein B0 corresponds to Glu.
7. The insulin analogue as claimed in claim 1 , wherein B1 corresponds to Asp.
8. The insulin analogue as claimed in claim 1 , wherein B2 corresponds to Val.
9. The insulin analogue as claimed in claim 1 , wherein B3 corresponds to Asp.
10. The insulin analogue as claimed in claim 1 , wherein B4 corresponds to Glu.
11. The insulin analogue as claimed in claim 1 , wherein B29 corresponds to Lys.
12. The insulin analogue as claimed in claim 1 , wherein B30 corresponds to Thr.
13. The insulin analogue as claimed in claim 1 , wherein B31 corresponds to Arg or Lys.
14. The insulin analogue as claimed in claim 1 , selected from the group consisting of:
Arg (A0), His (A8), Glu (A5), Gly (A21), Glu (B1), Arg (B31), Arg (B32)—NH 2 human insulin;
Arg (A0), His (A8), Glu (A5), Gly (A21), Glu (B1), Arg (B31), Lys (B32)—NH 2 human insulin;,
Arg (A0), His (A8), Glu (A15), Gly (A21), Glu (B1), Arg (B31), Arg (B32)—NH 2 human insulin;
Arg (A0), His (A8), Glu (A15), Gly (A21), Glu (B1), Arg (B31), Lys (B32)—NH 2 human insulin;
Arg (A0), His (A8), Asp (A18), Gly (A21), Glu (B1), Arg (B31), Arg (B32)—NH 2 human insulin;
Arg (A0), His (A8), Gly (A21), Glu (B1), Asp (B1), Arg (B31), Arg (B32)—NH 2 human insulin; and
Arg (A0), His (A8), Gly (A21), Glu (B1), Asp (B1), Arg (B31), Lys (B32)—NH 2 human insulin.
15. A process for preparing an insulin analogue as claimed in claim 1 , comprising the steps of:
(a) recombinantly preparing a precursor of the insulin analogue;
(b) enzymatically processing the precursor to two-chain insulin;
(c) coupling the two-chain insulin to argininamide in the presence of an enzyme having trypsin activity to produce the insulin analogue; and
(d) isolating the insulin analogue.
16. A method of making a medicament for treating diabetes mellitus, the method comprising combining the insulin analogue of claim 1 with one or more pharmaceutically acceptable ingredients.
17. The method of making of claim 16 , wherein the medicament is for the treatment of diabetes mellitus of type I or type II or for therapeutically assisting beta cell regeneration.
18. A pharmaceutical composition comprising the insulin analogue of claim 1 or a physiologically acceptable salt thereof.
19. A formulation of the insulin analogue of claim 1 , wherein the formulation is in aqueous form comprising the dissolved insulin analogue.
20. A formulation of the insulin analogue of claim 1 , wherein the formulation is in the form of a powder.
21. The formulation of claim 20 , wherein the insulin analogue is present in crystalline or amorphous form.
22. A formulation of the insulin analogue of claim 1 , wherein the formulation is in the form of a suspension.
23. A formulation of the insulin analogue of claim 1 , wherein the formulation further comprises a chemical chaperone.
24. The formulation of claim 19 , further comprises a glucagon-like peptide-1 (GLP1) or an analogue or derivative thereof, exendin-3 or an analogue or derivative thereof, or exendin-4 or an analogue or derivative therof.
25. The formulation of 20 , further comprises a glucagon-like peptide-1 (GLP1) or an analogue or derivative thereof, exendin-3 or an analogue or derivative thereof, or exendin-4 or an analogue or derivative thereof.
26. The formulation of claim 21 , further comprises a glucagon-like peptide-1 (GLP1) or an analogue or derivative thereof, exendin-3 or an analogue or derivative thereof, or exendin-4 or an analogue or derivative thereof.
27. The formulation of claim 22 , further comprises a glucagon-like peptide-1 (GLP1) or an analogue or derivative thereof, exendin-3 or an analogue or derivative thereof, or exendin-4 or an analogue or derivative thereof.
28. The formulation of claim 23 , further comprises a glucagon-like peptide-1 (GLP1) or an analogue or derivative thereof, exendin-3 or analogue or derivative thereof, or exendin-4 or an analogue or derivative thereof.
29. The formulation of claim 24 , wherein the formulation comprises exendin-4 or an analogue or derivative thereof.
30. The formulation of claim 29 , wherein the exendin-4 analogue is selected from a group comprising of:
H-desPro 36 -exendin-4-Lys 6 —NH 2 ,
H-des(Pro 36,37 )-exendin-4-Lys 4 —NH 2 , and
H-des(Pro 36,37 )-exendin-4-Lys 5 —NH 2 ,
or a pharmacologically tolerable salt thereof.
31. The formulation of claim 29 , wherein the exendin-4 analogue is selected from a group comprising of:
desPro 36 [Asp 28 ]exendin-4 (1-39),
desPro 36 [IsoAsp 28 ]exendin-4 (1-39),
desPro 36 [Met(O) 14 , Asp 28 ]exendin-4 (1-39),
desPro 36 [Met(O) 14 , IsoAsp 28 ]exendin-4 (1-39),
desPro 36 [Trp(O 2 ) 25 , Asp 28 ]exendin-4 (1-39),
desPro 36 [Trp(O 2 ) 25 , IsoAsp 28 ]exendin-4 (1-39),
desPro 36 [Met(O) 14 Trp(O 2 ) 25 , Asp 28 ]exendin-4 (1-39) and
desPro 36 [Met(O) 14 Trp(O 2 ) 25 , IsoAsp 28 ]exendin-4 (1-39),
or a pharmacologically tolerable salt thereof.
32. The formulation of claim 31 , where the peptide -Lys 6 —NH 2 is attached to the C terminus of the exendin-4 analogue.
33. The formulation of claim 24 , wherein the exendin-4 analogue is selected from the group comprising of:
H-(Lys) 6 -des Pro 36 [Asp 28 ]exendin-4(1-39)-Lys 6 -NH 2 ;
des Asp 28 Pro 36 , Pro 37 , Pro 38 exendin-4(1-39)—NH 2 ;
H-(Lys) 6 -des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]exendin-4(1-39)—NH 2 ;
H-Asn-(Glu) 5 des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]exendin-4(1-39)—NH 2 ;
des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]exendin-4(1-39)-(Lys) 6 —NH 2 ;
H-(Lys) 6 -des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]exendin-4(1-39)-(Lys) 6 —NH 2 ;
H-Asn-(Glu) 5 -des Pro 36 , Pro 37 , Pro 38 [Asp 28 ]exendin-4(1-39)-(Lys) 6 —NH 2 ;
H-(Lys) 6 -des Pro 36 [Trp(O 2 ) 25 , Asp 28 ]exendin-4(1-39)-Lys 6 —NH 2 ;
H-des Asp 28 Pro 36 , Pro 37 , Pro 38 [Trp(O 2 ) 25 ]exendin-4(1-39)—NH 2 ;
H-(Lys) 6 -des Pro 36 , Pro 37 , Pro 38 [TrP(O 2 ) 25 , Asp 28 ]exendin-4(1-39)—NH 2 ;
H-Asn-(Glu) 5 -des Pro 36 , Pro 37 , Pro 38 [Trp(O 2 ) 25 , Asp 28 ]exendin-4(1-39)—NH 2 ;
des Pro 36 , Pro 37 , Pro 38 [Trp(O 2 ) 25 , Asp 28 ]exendin-4(1-39)-(Lys) 6 —NH 2 ;
H-(Lys) 6 -des Pro 36 , Pro 37 , Pro 38 [TrP(O 2 ) 25 , Asp 28 ]exendin-4(1-39)-(Lys) 6 —NH 2 ;
H-Asn-(Glu) 5 -des Pro 36 , Pro 37 , Pro 38 [Trp(O 2 ) 25 , Asp 28 ]exendin-4(1-39)-(Lys) 6 —NH 2 ;
H-(Lys) 6 -des Pro 36 [Met(O) 14 , Asp 28 ]exendin-4(1-39)-Lys 6 —NH 2 ;
des Met(O) 14 Asp 28 Pro 36 , Pro 37 , Pro 38 exendin-4(1-39)—NH 2 ;
H-(Lys) 6 -des Pro 36 , Pro 37 , Pro 38 [Met(O) 14 , Asp 28 ]exendin-4(1-39)—NH 2 ;
H-Asn-(Glu) 5 -des Pro 36 , Pro 37 , Pro 38 [Met(O) 14 , Asp 28 ] exendin-4(1-39)—NH 2 ;
des Pro 36 , Pro 37 , Pro 38 [Met(O) 14 , Asp 28 ]exendin-4(1-39)-(Lys) 6 —NH 2 ;
H-(Lys) 6 -des Pro 36 , Pro 37 , Pro 38 [Met(O) 14 , Asp 28 ]exendin-4(1-39)-Lys 6 —NH 2 ;
H-Asn-(Glu) 5 des Pro 36 , Pro 37 , Pro 38 [Met(O) 14 , Asp 28 ] exendin-4(1-39)-(Lys) 6 —NH 2 ;
H-(Lys) 6 -des Pro 36 [Met(O) 14 , Trp(O 2 ) 25 , Asp 28 ]exendin-4(1-39)-Lys 6 —NH 2 ;
des Asp 28 Pro 36 , Pro 37 , Pro 38 [Met(O) 14 , Trp(O 2 ) 25 ]exendin-4(1-39)—NH 2 ;
H-(Lys) 6 -des Pro 36 ′ Pro 37 , Pro 38 [Met(O) 14 , Trp(O 2 ) 25 , Asp 28 ]exendin-4(1-39)—NH 2 ;
H-Asn-(Glu) 5 -des Pro 36 , Pro 37 , Pro 38 [Met(O) 14 , Asp 28 ] exendin-4(1-39)—NH 2 ;
des Pro 36 , Pro 37 , Pro 38 [Met(O) 14 , Trp(O 2 ) 25 , Asp 28 ]exendin-4(1-39)-(Lys) 6 —NH 2 ;
H-(Lys) 6 -des Pro 36 ′ Pro 37 , Pro 38 [Met(O) 14 , Trp(O 2 ) 25 , Asp 28 ]exendin-4(1-39)-(Lys) 6 —NH 2 ; and
H-Asn-(Glu) 5 -des Pro 36 , Pro 37 , Pro 38 [Met(O) 14 , Trp(O 2 ) 25 , Asp 28 ] exendin-4(1-39) -(Lys) 6 —NH 2 ;
or a pharmacologically tolerable salt thereof.
34. The formulation of claim 24 , further comprising Arg 34 , Lys 26 (N ε (γ-glutamyl(N α -hexadecanoyl))) GLP-1 (7-37) [liraglutide] or a pharmacologically tolerable salt thereof.
35. An aqueous formulation comprising the insulin analogue of claim 1 , which comprises less than 15 μg/ml of zinc.
36. An aqueous formulation comprising the insulin analogue of claim 1 , which comprises less than 2 mg/ml of zinc.
37. The aqueous formulation of claim 36 , which comprises 200 μg/ml of zinc.