CLAIMS
1. An acylated analogue of human insulin, which analogue is [A22Lys, BSaar1] relative to human insulin; wherein
aar1 represents Glu (E), Gin (Q), Asp (D), Ser (S) orThr (T); and
one, two or three of the amino acid residues located in positions B26, B27 and/or B28 are substituted as follows:
the amino acid residue located in position B26 is substituted for Glu (E) or Asp (D); and/or
the amino acid residue located in position B27 is substituted for Glu (E) Asp (D), or Pro (P); and/or
the amino acid residue located in position B28 is substituted for Glu (E), Asp (D) or Arg (R); provided, however;
if the amino acid residue located in position B27 is substituted for Pro (P); then the amino acid residue located in position B28 is substituted for Glu (E), Asp (D) or Arg (R); and
if the amino acid residue located in position B28 is not substituted, then the amino acid residue located in position B29 is substituted for B29Arg (R), or
if the amino acid residue located in position B28 is substituted for Glu (E) or Asp (D), then the amino acid residue located in position B29 is substituted for B29Pro (P) or B29Arg (R), and
if the amino acid residue located in position B29 is Pro (P), then the amino acid residue located in position B30 is substituted for B30Arg (R); and
if the amino acid residue located in position B28 is substituted for Arg (R), then the amino acid residues located in positions B29 and B30 have been deleted (i.e. desB29, desB30); and
if the amino acid residue located in position B29 is substituted for Arg (R), then the amino acid residue located in position B30 has been deleted (i.e. desB30); and
which analogue may additionally comprise an A8Arg (R) and/or an A14Glu (E) substitution; and
which insulin analogue is derivatized by acylation of the epsilon amino group of the lysine residue at the A22 position with a group of Formula II
[Acyl]-[Linker]-
wherein the Linker group is an amino acid chain composed of from 1 to 10 amino acid residues selected from gGlu and/or OEG; wherein
gGlu represents a gamma glutamic acid residue;

OEG represents a residue of 8-amino-3,6-dioxaoctanoic acid (i.e. a group of the
formula -NH-(CH2)2-0-(CH2)2-0-CH2-CO-);
which amino acid residues may be present in any order; and
which amino acid chain comprises at least one gGlu residue; and
wherein the Acyl group is a residue of an a,co-di-carboxylic acid selected from
1,14-tetradecanedioic acid; 1,15-pentadecanedioic acid; and 1,16-hexadecanedioic acid.
2. The acylated insulin analogue according to claim 1, which analogue is
[A22Lys; BSaar1; B29Arg; desB30] relative to human insulin; wherein
aar1 represents Glu (E), Gin (Q), Asp (D), Ser (S) orThr (T); and one, two or three of the amino acid residues located in positions B26, B27 and/or B28 are substituted with Glu (E) or Asp (D).
3. The acylated insulin analogue according to claim 1, which analogue is
[A22Lys; BSaar1; B26aar2; B29Arg; desB30] relative to human insulin; wherein
aar1 represents Glu (E), Gin (Q), Asp (D), Ser (S) orThr (T); and aar2 represents Glu (E) or Asp (D).
4. The acylated insulin analogue according to claim 1, which analogue is
[A8R, A22K, B3E, B26E, B27E, B28R, desB29, desB30];
[A8R, A22K, B3E, B26E, B27P, B28R, desB29, desB30]; [A8R, A22K, B3E, B26E, B28E, B29R, desB30]; [A8R, A22K, B3E, B26E, B29R, desB30]; [A8R, A22K, B3E, B27E, B28E, B29R, desB30]; [A8R, A22K, B3E, B27E, B28R, desB29, desB30]; [A14E, A22K, B3E, B27E, B28E, B29R, desB30]; [A14E, A22K, B3Q, B27E, B28E, B29R, desB30]; [A14E, A22K, B3Q, B28D, B29R, desB30]; [A22K, B3E, B26E, B27E, B28E, B29R, desB30]; [A22K, B3E, B26E, B27E, B28R, desB29, desB30]; [A22K, B3E, B26E, B27E, B29P, B30R]; [A22K, B3E, B26E, B27P, B28R, desB29, desB30]; [A22K, B3E, B26E, B28E, B29P, B30R]; [A22K, B3E, B26E, B28E, B29R, desB30]; [A22K, B3E, B26E, B28D, B29R, desB30]; [A22K, B3E, B26E, B29R, desB30];

[A22K, B3E, B27E, B28E, B29P, B30R]; [A22K, B3E, B27E, B28E, B29R, desB30]; [A22K, B3E, B27E, B28R, desB29, desB30]; [A22K, B3E, B27E, B29R, desB30]; [A22K, B3E, B27P, B28E, B29R, desB30]; [A22K, B3E, B28D, B29R, desB30]; [A22K, B3E, B28E, B29P, B30R]; [A22K, B3Q, B26E, B28E, B29R, desB30]; or [A22K, B3Q, B26E, B29R, desB30]; relative to human insulin.
5. The acylated insulin analogue according to any one of claims 1-4, wherein, in
the group of Formula II
[Acyl]-[l_inker]-the Linker group is an amino acid chain composed of from 1 to 10 amino acid residues selected from gGlu and/or OEG; which amino acid residues may be present in any order; and which amino acid chain comprises at least one gGlu residue.
6. The acylated insulin analogue according to any one of claims 1-4, wherein, in
the group of Formula II
[Acyl]-[l_inker]-the Acyl group is a residue of an a,co-di-carboxylic acid selected from 1,14-tetradecanedioic acid; 1,15-pentadecanedioic acid; and 1,16-hexadecanedioic acid.
7. The acylated insulin analogue according to any one of claims 1-4, wherein the
group of Formula II is
tetradecanedioyl-4xgGlu;
tetradecanedioyl-gGlu-2xOEG;
hexadecanedioyl-4xgGlu;
hexadecanedioyl-4xgGlu-2xOEG;
hexadecanedioyl-gGlu-2xOEG;
hexadecanedioyl-gGlu-4xOEG; or
hexadecanedioyl-gGlu-6xOEG.
8. The acylated insulin analogue according to claim 1, which is

A22K(N(eps)tetradecanedioyl-gGlu-2xOEG), B3E, B27E, B28E, B29R, desB30 human insulin;
A14E, A22K(N(eps)tetradecanedioyl-gGlu-2xOEG), B3E, B27E, B28E, B29R, desB30 human insulin;
A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B27E, B28E, B29R, desB30 human insulin;
A14E, A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B27E, B28E, B29R, desB30 human insulin;
A22K(N(eps)tetradecanedioyl-gGlu-2xOEG), B3E, B27P, B28E, B29R, desB30 human insulin;
A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B27E, B29R, desB30 human insulin;
A22K(N(eps)tetradecanedioyl-gGlu-2xOEG), B3E, B28D, B29R, desB30 human insulin;
A14E, A22K(N(eps)tetradecanedioyl-4xgGlu), B3Q, B28D, B29R, desB30 human insulin;
A14E, A22K(N(eps)tetradecanedioyl-4xgGlu), B3Q, B27E, B28E, B29R, desB30 human insulin;
A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B28D, B29R, desB30 human insulin;
A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B26E, B28E, B29R, desB30 human insulin;
A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B26E, B29R, desB30 human insulin;
A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B26E, B27P, B28R, desB29, desB30 human insulin;
A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B27E, B28R, desB29, desB30 human insulin;
A8R, A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B26E, B29R, desB30 human insulin;
A8R, A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B26E, B27P, B28R, desB29, desB30 human insulin;
A8R, A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B27E, B28R, desB29, desB30 human insulin;
A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B26E, B27E, B28R, desB29, desB30 human insulin;
A8R, A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B26E, B27E, B28R, desB29, desB30 human insulin;
A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B28E, B29P, B30R human insulin;

A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B26E, B28E, B29P, B30R human insulin;
A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B27E, B28E, B29P, B30R human insulin;
A8R, A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B26E, B28E, B29R, desB30 human insulin;
A8R, A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B27E, B28E, B29R, desB30 human insulin;
A22K(N(eps)Hexadecanedioyl-gGlu-2xOEG), B3E, B27E, B29R, desB30 human insulin;
A14E, A22K(N(eps)hexadecanedioyl-gGlu-2xOEG), B3E, B27E, B28E, B29R, desB30 human insulin;
A14E, A22K(N(eps)hexadecanedioyl-4xgGlu), B3E, B27E, B28E, B29R, desB30 human insulin;
A22K(N(eps)hexadecanedioyl-4xgGlu), B3E, B27E, B28E, B29R, desB30 human insulin;
A22K(N(eps)hexadecanedioyl-gGlu-2xOEG), B3E, B27P, B28E, B29R, desB30 human insulin;
A22K(N(eps)hexadecanedioyl-4xgGlu), B3E, B27P, B28E, B29R, desB30 human insulin;
A22K(N(eps)hexadecanedioyl-gGlu-2xOEG), B3E, B27E, B28E, B29R, desB30 human insulin;
A22K(N(eps)hexadecanedioyl-4xgGlu), B3E, B27E, B29R, desB30 human insulin;
A22K(N(eps)hexadecanedioyl-gGlu-2xOEG), B3E, B28D, B29R, desB30 human insulin;
A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B26E, B28D, B29R, desB30 human insulin;
A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B26E, B27E, B29P, B30R human insulin;
A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B26E, B27E, B28E, B29R, desB30 human insulin;
A22K(N(eps)hexadecanedioyl-4xgGlu), B3E, B28D, B29R, desB30 human insulin;
A22K(N(eps)tetradecanedioyl-4xgGlu), B3E, B27P, B28E, B29R, desB30 human insulin;
A22K(N(eps)tetradecanedioyl-4xgGlu), B3Q, B26E, B29R, desB30 human insulin;

A22K(N(eps)tetradecanedioyl-4xgGlu), B3Q, B26E, B28E, B29R, desB30 human insulin;
A22K(N(eps)hexadecanedioyl-4xgGlu), B3E, B26E, B29R, desB30 human insulin;
A22K(N(eps)hexadecanedioyl-gGlu-2xOEG), B3E, B26E, B29R, desB30 human insulin;
A22K(N(eps)hexadecanedioyl-gGlu-4xOEG), B3E, B26E, B29R, desB30 human insulin;
A22K(N(eps)hexadecanedioyl-gGlu-6xOEG), B3E, B26E, B29R, desB30 human insulin;
A22K(N(eps)hexadecanedioyl-4xgGlu-2xOEG), B3E, B26E, B29R, desB30 human insulin; or
A22K(N(eps)hexadecanedioyl-4xgGlu), B3Q, B26E, B29R, desB30 human insulin.
9. A pharmaceutical composition comprising an insulin derivative according to
any one of claims 1-8, and one or more pharmaceutical^ acceptable carriers or diluents.
10. The pharmaceutical composition according to claim 9, formulated as a low-zinc composition, with no added zinc ions.
11. The pharmaceutical composition according claim 10, formulated as a low-zinc composition, comprising less than 0.2 Zn2+ ions per 6 insulin molecules.
12. The low-zinc pharmaceutical composition according to either one of claims 10-11, wherein no surfactant has been added.
13. The low-zinc pharmaceutical composition according to any one of claims 10-12, comprising a nicotinic compound, and in particular nicotinamide.
14. The insulin analogue according to any one of claims 1-8, or a
pharmaceutically acceptable salt thereof, for use as a medicament.
15. A method of treatment, prevention or alleviation of a metabolic disease or
disorder or condition of a living animal body, including a human, which method comprises
the step of administering to such a living animal body in need thereof, a therapeutically
effective amount of the acylated insulin analogue according to any one of claims 1-8.