INSULIN ANALOGUES WITH GLUCOSE REGULATED CONFORMATIONAL SWITCH
The present invention relates to glucose-responsive insulin analogues, compositions including the glucose-responsive insulin analogues, and methods of lowering blood sugar of a patient using the insulin analogue or compositions thereof.
1 . A glucose-responsive insulin analogue or complex, comprising:
an insulin analogue,
one or more glucose-binding moieties, and
one or more binding partner moieties
wherein by interaction of the glucose-binding moiety and binding partner moiety the glucose-responsive insulin analogue or complex has low affinity for the insulin receptor at euglycemic concentrations of glucose and has increased affinity for the insulin receptor by disruption of the glucose-binding moiety and binding partner moiety interaction at hyperglycemic concentrations of glucose.
2 . The insulin analogue or complex of claim 1 , wherein the glucose-binding moiety is a multivalent boronic acid.
3 . The insulin analogue or complex of claim 2 wherein the multivalent boronic acid moiety comprises one or more aromatic boronic acids, which is optionally a halogen-modified aromatic boronic acid.
4 . The insulin analogue or complex of claim 2 or 3 , wherein the multivalent boronic acid moiety is a divalent boronic acid moiety.
5 . The insulin analogue or complex of claim 4 , wherein the divalent boronic acid moiety further comprises a linker, which is optionally an amide linker, an acyl linker, or a PEG linker.
6 . The insulin analogue or complex of claim 5 , wherein the divalent boronic acid moiety is a compound of Formula (I):
wherein:
X is 1, 2, 3, 4, or 5, and
R and/or R′ comprises a group independently selected from
where X is selected from F, Cl, Br, I, —NH 2 , —OMe, —NO 2 ;
where Y is selected from F, Cl, Br, I, —NH 2 , —OMe, —NO 2 , —NH(CH 3 ), —N(CH 3 ) 2 , —(CH 2 ) a NH(CH 3 ), or —N(CH 2 ) b (CH 3 ) 2 ; a=1, 2, or 3 and b=1, 2, or 3;
where Z is selected from F, Cl, Br, I, —NH 2 , —OMe, —NO 2 ; and
where Y is selected from F, Cl, Br, I, —NH 2 , —OMe, —NO 2 , —NH(CH 3 ), —N(CH 3 ) 2 , —(CH 2 ) a NH(CH 3 ), or —N(CH 2 ) b (CH 3 ) 2 ; a=1, 2, or 3 and b=1, 2, or 3.
7 . The insulin analogue or complex of claim 3 , wherein the divalent boronic acid moiety has the following structure:
8 . The insulin analogue or complex of claim 5 , wherein the divalent boronic acid moiety has the following formula:
9 . The insulin analogue or complex of claim 5 , wherein the divalent boronic acid moiety is a compound of Formula (II):
wherein:
X is 1-10, and
R and/or R′ comprises a group independently selected from
where X is selected from F, Cl, Br, I, —NH 2 , —OMe, —NO 2 ;
where Y is selected from F, Cl, Br, I, —NH 2 , —OMe, —NO 2 , —NH(CH 3 ), —N(CH 3 ) 2 , —(CH 2 ) a NH(CH 3 ), or —N(CH 2 ) b (CH 3 ) 2 ; a=1, 2, or 3 and b=1, 2, or 3;
where Z is selected from F, Cl, Br, I, —NH 2 , —OMe, —NO 2 ; and
where Y is selected from F, Cl, Br, I, —NH 2 , —OMe, —NO 2 , —NH(CH 3 ), —N(CH 3 ) 2 , —(CH 2 ) a NH(CH 3 ), or —N(CH 2 ) b (CH 3 ) 2 ; a=1, 2, or 3 and b=1, 2, or 3.
10 . The insulin analogue or complex of claim 5 , wherein the divalent boronic acid moiety has a structure of Formula (III):
wherein:
X is 1-10, and
R and/or R′ comprises a group independently selected from
where X is selected from F, Cl, Br, I, —NH 2 , —OMe, —NO 2 ;
where Y is selected from F, Cl, Br, I, —NH 2 , —OMe, —NO 2 , —NH(CH 3 ), —N(CH 3 ) 2 , —(CH 2 ) a NH(CH 3 ), or —N(CH 2 ) b (CH 3 ) 2 ; a=1, 2, or 3 and b=1, 2, or 3;
where Z is selected from F, Cl, Br, I, —NH 2 , —OMe, —NO 2 ; and
where Y is selected from F, Cl, Br, I, —NH 2 , —OMe, —NO 2 , —NH(CH 3 ), —N(CH 3 ) 2 , —(CH 2 ) a NH(CH 3 ), or —N(CH 2 ) b (CH 3 ) 2 ; a=1, 2, or 3 and b=1, 2, or 3.
11 . The insulin analogue or complex of any one of claims 1 to 10 , wherein the binding partner is a polyol-containing moiety.
12 . The insulin analogue or complex of claim 11 , wherein the polyol-containing moiety is a sugar mimetic.
13 . The insulin analogue or complex of claim 11 , wherein the polyol-containing moiety is natural or synthetically modified monosaccharide, disaccharide, and/or an oligosaccharide.
14 . The insulin analogue or complex of any one of claims 1 to 13 , wherein the glucose-binding moiety and/or the binding partner moiety are amine-linked or thiol-linked to the insulin analogue.
15 . The insulin analogue or complex of claim 14 , wherein the glucose binding moiety and/or the binding partner moiety are linked to an alpha-amino functional group or a side chain amino functional group of an amino acid residue of the insulin analogue.
16 . The insulin analogue or complex of claim 15 , wherein one or more amino acids of the insulin analogue are substituted with an amino acid selected from D-homolysine, D-lysine, D-ornithine, D-diaminobutyric acid, D-diaminopropionic acid, L-homolysine, L-lysine, L-ornithine, L-diaminobutyric acid, and L-diaminopropionic acid, which optionally are linked to the glucose-binding moiety and/or the binding partner moiety.
17 . The insulin analogue or complex of any one of claims 1 to 16 , wherein the glucose-binding moiety and/or the binding partner moiety is linked to the insulin analogue via a spacer.
18 . The insulin analogue or complex of claim 17 , wherein the spacer comprises an amide linker, an acyl linker, or a PEG linker.
19 . The insulin analogue or complex of claim 18 , wherein the spacer is an acyl linker with 1 to 16 carbon atoms, optionally, with one more nitrogen atoms at or near the end of the linker.
20 . The insulin analogue or complex of any one of claims 1 to 19 , wherein the insulin analogue comprises an A-chain polypeptide and a B-chain polypeptide and wherein:
the A-chain is modified with a glucose-binding moiety and the B-chain is modified with a binding partner moiety; or
the B-chain is modified with a glucose-binding moiety and the A-chain is modified with a binding partner moiety.
21 . The insulin analogue or complex of claim 20 , wherein the glucose-binding moiety is attached at or near N-terminus of the A-chain and the binding partner moiety is attached at or near C-terminus of the B-chain.
22 . The insulin analogue or complex of claim 20 , wherein the glucose-binding moiety is attached at or near C-terminus of the B-chain and the binding partner moiety is attached at or near N-terminus of the A-chain.
23 . The insulin analogue or complex of claim 21 , wherein the glucose-binding moiety is attached to a position selected from A1-A10.
24 . The insulin analogue or complex of claim 23 , wherein the binding partner moiety is attached to a position selected from B16-B30.
25 . The insulin analogue or complex of claim 23 , wherein the binding partner moiety is linked to a B-chain extension, wherein the extension has an additional B31 amino acid or additional B31 and B32 amino acids.
26 . The insulin analogue or complex of claim 22 , wherein the glucose-binding moiety is attached to a position selected from B16-B30.
27 . The insulin analogue or complex of claim 26 , wherein the binding partner moiety is attached to a position selected from A1-A10.
28 . The insulin analogue or complex of claim 26 , wherein the amino acid at position B31 is a cysteine or a homocysteine and the amino acid at position B32 is cysteine or a homocysteine.
29 . The insulin analogue or complex of any one of claims 1 to 20 wherein the insulin analogue comprises an A-chain polypeptide and a B-chain polypeptide and wherein:
the A-chain is modified with a glucose-binding moiety or binding partner moiety, and the B-chain is modified with a glucose-binding moiety or binding partner moiety, and wherein the glucose-binding moiety or binding partner moiety of the A chain and the glucose-binding moiety or binding partner moiety of the B chain are bound by cognate binding partners attached through a linker.
30 . The insulin analogue or complex of claim 29 , wherein a glucose-binding moiety or binding partner moiety of the A chain is attached at or near the N-terminus of the A-chain; and the glucose-binding moiety or binding partner moiety of the B chain is attached at or near C-terminus of the B-chain.
31 . The insulin analogue or complex of claim 30 , wherein the glucose-binding moiety or binding partner moiety of the A chain is attached to a position selected from A1-A10.
32 . The insulin analogue or complex of claim 31 , wherein the glucose-binding moiety or the binding partner moiety of the B chain is attached to a position selected from B16-B30.
33 . The insulin analogue or complex of any one of claims 29 to 32 , wherein a glucose binding moiety or binding partner moiety is linked to a B-chain extension, wherein the extension has an additional B31 amino acid or additional B31 and B32 amino acids.
34 . The insulin analogue or complex of claim 33 , wherein the amino acid at position B31 is a cysteine or a homocysteine and the amino acid at position B32 is cysteine or a homocysteine.
35 . The insulin analogue or complex of any one of claims 1 to 19 , wherein the insulin analogue comprises an A-chain polypeptide and a B-chain polypeptide and wherein:
the A-chain is modified with a glucose-binding moiety and binding partner moiety each coupled to the A-chain through a spacer, or the B-chain is modified with a glucose-binding moiety and a binding partner moiety each coupled to the B-chain through a spacer.
36 . The insulin analogue or complex of claim 35 , wherein the glucose-binding moiety and binding partner moiety are attached at or near the N-terminus of the A-chain; or the glucose-binding moiety and the binding partner moiety are attached at or near C-terminus of the B-chain.
37 . The insulin analogue or complex of claim 36 , wherein the glucose-binding moiety and the binding partner moiety are attached to a position selected from A1-A10.
38 . The insulin analogue or complex of claim 36 , wherein the glucose-binding moiety and the binding partner moiety are attached to a position selected from B16-B30.
39 . The insulin analogue or complex of claim 36 , wherein a glucose binding moiety or binding partner moiety is linked to a B-chain extension, wherein the extension has an additional B31 amino acid or additional B31 and B32 amino acids.
40 . The insulin analogue or complex of claim 39 , wherein the amino acid at position B31 is a cysteine or a homocysteine and the amino acid at position B32 is cysteine or a homocysteine.
41 . The insulin analogue or complex of any one of claims 1 to 19 , wherein the insulin analogue comprises an A-chain polypeptide and a B-chain polypeptide and wherein:
the A-chain and/or the B chain is modified with a glucose-binding moiety or a binding partner moiety, and the cognate binding partner is bound to the glucose-binding moiety or binding partner moiety so as to inhibit binding of the insulin analogue to the insulin receptor.
42 . The insulin analogue or complex of claim 41 , wherein the cognate binding partner conjugated to a macromolecular carrier optionally through a spacer.
43 . The insulin of claim 42 , wherein the macromolecular carrier is albumin or an albumin-binding molecule.
44 . The insulin analogue or complex of any one of claims 41 to 43 , wherein a glucose-binding moiety or a binding partner moiety are attached at or near the N-terminus of the A-chain; and/or a glucose-binding moiety and/or a binding partner moiety are attached at or near C-terminus of the B-chain.
45 . The insulin analogue or complex of claim 44 , wherein the glucose-binding moiety or the binding partner moiety are attached to a position selected from A1-A10.
46 . The insulin analogue or complex of claim 44 , wherein the glucose-binding moiety or the binding partner moiety are attached to a position selected from B16-B30.
47 . The insulin analogue or complex of any one of claims 41 to 46 , wherein a glucose binding moiety or binding partner moiety is linked to a B-chain extension, wherein the extension has an additional B31 amino acid or additional B31 and B32 amino acids.
48 . The insulin analogue or complex of claim 47 , wherein the amino acid at position B31 is a cysteine or a homocysteine and the amino acid at position B32 is cysteine or a homocysteine.
49 . The insulin analogue or complex of any one of claims 1 to 48 , where the insulin is a mammalian insulin or a variant thereof.
50 . A composition comprising the insulin analogue or complex of any one of claims 1 to 48 and a pharmaceutically acceptable carrier or excipient.
51 . The composition of claim 50 , wherein the composition is formulated for subcutaneous delivery.
52 . A method of lowering the blood sugar of a patient comprising a physiologically effective amount of the insulin analogue or complex according to any one of claims 1 to 48 or a composition according to claim 50 or 51 .