INSULIN COMPOSITIONS AND METHOD OF MAKING A COMPOSITION

• US 20090074882A1
• Filed: 12/21/2006
• Published: 03/19/2009
• Est. Priority Date: 12/28/2005
• Status: Abandoned Application

• Current Assignee: Novo Nordisk A/S
• Original Assignee: Novo Nordisk A/S
• Inventors: Olsen, Helle Birk, Plum, Anne, Havelund, Svend, Hoeg-Jensen, Thomas, Ribel-Madsen, Ulla, Hubalek, Frantisek, Jonassen, Ib
• Application Number:
  US12/096,476
• Publication Number:
  US 20090074882A1
• Time in Patent Office: Days
• US Class Current: 424/641
• CPC Class Codes: A61K 38/28 Insulins
• CPC Class Codes: A61K 47/52 the modifying agent being an inorganic compound, e.g. an inorganic ion that is complexed with the active ingredient
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C07K 14/62 Insulins

Abstract

The invention is related to insulin compositions with a high content of zinc atoms per six molecules of acylated insulin. The insulin is an acylated insulin and may be mixed with a further insulin analogue such as the rapid acting insulin Asp B28 human insulin.

Claims

1. A soluble pharmaceutical composition comprising an acylated insulin and further comprising more than 4 zinc atoms per 6 molecules of acylated insulin.

2. The pharmaceutical composition according to claim 1 comprising up to about 12 zinc atoms per 6 molecules of acylated insulin.

3. The pharmaceutical composition according to claim 1 comprising between about 4.3 and about 12 zinc atoms per 6 molecules of acylated insulin.

4. The pharmaceutical composition according to claim 1 comprising between about 4.5 and about 12 zinc atoms per 6 molecules of acylated insulin.

5. The pharmaceutical composition according to claim 1, wherein at least 85% of the acylated insulin is present as complexes which are acylated insulin dodecamers or complexes with a higher molecular weight than acylated insulin dodecamer.

6. The pharmaceutical composition according to claim 1, wherein the composition comprises a surfactant.

7. The pharmaceutical composition according to claim 1, wherein the insulin molecule has a side chain attached either to the α

-amino group of the N-terminal amino acid residue of the B chain or to an Eε

-amino group of a Lys residue present in the B chain of the parent insulin moiety via an amide bond, which side chain comprises at least one free carboxylic acid group or a group which is negatively charged at neutral pH, a fatty acid moiety with about 4 to about 32 carbon atoms in the carbon chain; and

possible one or more linkers linking the individual components in the side chain together via amide bonds.

8. The pharmaceutical composition according to claim 1, wherein the side chain comprises at least one aromatic group.

9. The pharmaceutical composition according to claim 1, wherein the side chain comprises at least one difunctional PEG group.

10. The pharmaceutical composition according to claim 1, wherein the insulin molecule has a side chain attached to the Eε

-amino group of a Lys residue present in the B chain of the parent insulin, the side chain being of the general formula;

—

W—

X—

Y-Z₂

wherein W is;

an α

-amino acid residue having a carboxylic acid group in the side chain which residue forms, with one of its carboxylic acid groups, an amide group together with ε

-amino group of a Lys residue present in the B chain of the parent insulin;

a chain composed of two, three or four α

-amino acid residues linked together via amide carbonyl bonds, which chain—

via an amide bond—

is linked to an ε

-amino group of a Lys residue present in the B chain of the parent insulin, the amino acid residues of W being selected from the group of amino acid residues having a neutral side chain and amino acid residues having a carboxylic acid group in the side chain so that W has at least one amino acid residue which has a carboxylic acid group in the side chain;

or

a covalent bond from X to an ε

-amino group of a Lys residue present in the B chain of the parent insulin;

X is;

—

CO—

;

—

CH(COOH)CO—

;

—

CO—

N(CH₂COOH)CH₂CO—

;

—

CO—

N(CH₂COOH)CH₂CON(CH₂COOH)CH₂CO—

;

—

CO—

N(CH₂CH₂COOH)CH₂CH₂CO—

;

—

CO—

N(CH₂CH₂COOH)CH₂CH₂CON(CH₂CH₂COOH)CH₂CH₂CO—

;

—

CO—

NHCH(COOH)(CH₂)₄NHCO—

;

—

CO—

N(CH₂CH₂COOH)CH₂CO—

;

or

—

CO—

N(CH₂COOH)CH₂CH₂CO—

.

that

a) when W is an amino acid residue or a chain of amino acid residues, via a bond from the underscored carbon forms an amide bond with an amino group in W, or

b) when W is a covalent bond, via a bond from the underscored carbonyl carbon forms an amide bond with an ε

-amino group of a Lys residue present in the B chain of the parent insulin;

Y is;

—

(CH₂)_m—

where m is an integer in the range of 6 to 32;

a divalent hydrocarbon chain comprising 1, 2 or 3 —

CH═

CH—

groups and a number of —

CH₂—

groups sufficient to give a total number of carbon atoms in the chain in the range of 10 to 32; and

Z₂ is;

—

COOH;

—

CO-Asp;

—

CO-Glu;

—

CO-Gly;

—

CO-Sar;

—

CH(COOH)₂;

—

N(CH₂COOH)₂;

—

SO₃H;

or

—

PO₃H

and any Zn²⁺ complexes thereof, provided that when W is a covalent bond and X is —

CO—

, then Z is different from —

COOH.

11. The pharmaceutical composition according to claim 1, wherein the acylated insulin is having a formula

wherein Ins is the parent insulin moiety which via the α

-amino group of the N-terminal amino acid residue of the B chain or an ε

-amino group of a Lys residue present in the B chain of the insulin moiety is bound to the CO—

group in the side chain via an amide bond;

X₄ is

—

(CH₂)_n where n is 1, 2, 3, 4, 5 or 6;

NR, where R is hydrogen or —

(CH₂)_p—

COOH;

—

(CH₂)_p—

SO₃H;

—

(CH₂)_p—

PO₃H₂;

—

(CH₂)_p—

O—

SO₃H₂;

—

(CH₂)_p—

O—

PO₃H₂;

arylene substituted with 1 or 2 —

(CH₂)_p—

O—

COOH groups;

—

(CH₂)_p-tetrazolyl, where p is an integer in the range of 1 to 6;

—

(CR₁R₂)_q—

NR—

CO—

, where R₁ and R₂ independently of each other and independently for each value of q can be H, —

COOH, or OH, q is 1-6 and R is defined as above;

—

((CR₃R₄)_q1—

NR—

CO)_2-4—

, where R₃ and R₄ independently of each other and independently for each value of q₁ can be H, —

COOH, or OH, q₁ is 1-6 and R is defined as above;

or

a bond

W₁ is arylene or heteroarylene, which may be substituted with one or two groups selected from the group consisting of —

COOH, —

SO₃H, and —

PO₃H₂ and tetrazolyl, or W₁ is a bond;

m is 0, 1, 2, 3, 4, 5 or 6;

X₅ is

where R is defined as above;

or

a bond;

Y₁ is

—

(CR₁R₂)_q—

NR—

CO—

, where R₁ and R₂ independently of each other and independently for each value of q can be H, —

COOH, a bond or OH, q is 1-6; and

R is defined as above;

NR where R is defined as above;

—

((CR₃R₄)_q1—

NR—

CO)_2-4—

, where R₃ and R₄ independently of each other and independently for each value of q₁ can be H, —

COOH, or OH, q₁ is 1-6 and R is defined as above;

or

a bond;

Q₇ is

—

(CH₂)_r—

where r is an integer from 4 to 22;

a divalent hydrocarbon chain comprising 1, 2 or 3 —

CH═

CH—

groups and a number of —

CH₂—

groups sufficient to give a total number of carbon atoms in the chain in the range of 4 to 22;

or

a divalent hydrocarbon chain of the formula

—

(CH₂)_s-Q₈-(C₆H₄)_v1-Q₉-(CH₂)_W-Q₁₀-(C₆H₄)_v2-Q₁₁-(CH₂)_t-Q₁₂-(C₆H₄)_v3-Q₁₃-(CH₂)_z—

wherein Q₈-Q₁₃ independently of each other can be O;

S or a bond;

where s, w, t and z independently of each other are zero or an integer from 1 to 10 so that the sum of s, w, t and z is in the range from 4 to 22, and v₁, v₂, and v₃ independently of each other can be zero or 1, provided that when W₁ is a bond then Q₇ is not a divalent hydrocarbon chain of the formula —

(CH₂)_v4C₆H₄(CH₂)_W1—

wherein v₄ and w₁ are integers or one of them is zero so that the sum of v₄ and w₁ is in the range of 6 to 22; and

Z₁ is;

—

COOH;

—

CO-Asp;

—

CO-Glu;

—

CO-Gly;

—

CO-Sar;

—

CH(COOH)₂;

—

N(CH₂COOH)₂;

—

SO₃H

—

PO₃H₂;

—

O—

SO₃H;

—

O—

PO₃H₂;

-tetrazolyl or

—

O—

W₂,

where W₂ is arylene or heteroarylene substituted with one or two groups selected from —

COOH, —

SO₃H, and —

PO₃H₂ and tetrazolyl;

provided that if W₁ is a bond and v₁, v₂ and v₃ are all zero and Q₈-13 are all a bonds, then Z₁ is O—

W₂

and any Zn²⁺ complex thereof.

12. The pharmaceutical composition to claim 1, wherein the acylated insulin is having a formula

wherein Ins is the parent insulin moiety which via the α

-amino group of the N-terminal amino acid residue of the B chain or an ε

-amino group of a Lys residue present in the B chain of the insulin moiety is bound to the CO—

group in the side chain via an amide bond;

each n is independently 0, 1, 2, 3, 4, 5 or 6;

Q₁, Q₂, Q₃, and Q₄ independently of each other can be

(CH₂CH₂O)_s—

;

(CH₂CH₂CH₂O)_s—

;

(CH₂CH₂CH₂CH₂O)_s—

;

(CH₂CH₂OCH₂CH₂CH₂CH₂O)_s—

or (CH₂CH₂CH₂OCH₂CH₂CH₂CH₂O)_s—

where s is 1-20

—

(CH₂)_r—

where r is an integer from 4 to 22;

or a divalent hydrocarbon chain comprising 1, 2 or 3 —

CH═

CH—

groups and a number of —

CH₂—

groups sufficient to give a total number of carbon atoms in the chain in the range of 4 to 22;

—

(CH₂)_t—

or —

(CH₂OCH₂)_t—

, where t is an integer from 1 to 6;

—

(CR₁R₂)_q—

, where R₁ and R₂ independently of each other can be H, —

COOH, (CH₂)_1-6COOH and R₁ and R₂ can be different at each carbon, and q is 1-6,

—

((CR₃R₄)_q1)₁—

(NHCO—

(CR₃R₄)_q1—

NHCO)_1-2—

((CR₃R₄)_q1)₁ or ((CR₃R₄)_q1)₁—

(CONH—

(CR₃R₄)_q1—

CONH)_1-2—

((CR₃R₄)_q1—

)—

, —

((CR₃R₄)_q1)₁—

(NHCO—

(CR₃R₄)_q1—

CONH)_1-2—

((CR₃R₄)_q1)₁ or —

((CR₃R₄)_q1)₁—

(CONH—

(CR₃R₄)_q1—

NHCO)_1-2—

((CR₃R₄)_q1)₁ where R₃ and R₄ independently of each other can be H, —

COOH, and R₃ and R₄ can be different at each carbon, and q₁ is 1-6-, or

a bond;

with the proviso that Q₁-Q₄ are different;

X₁, X₂ and X₃ are independently

O;

a bond;

or

where R is hydrogen or —

(CH₂)_p—

COOH, —

(CH₂)_p—

SO₃H, —

(CH₂)_p—

PO₃H₂, —

(CH₂)_p—

O—

SO₃H;

—

(CH₂)_p—

O—

PO₃H₂;

or —

(CH₂)_p-tetrazol-5-yl, where each p independently of the other p'"'"'s is an integer in the range of 1 to 6; and

Z is;

—

COOH;

—

CO-Asp;

—

CO-Glu;

—

CO-Gly;

—

CO-Sar;

—

CH(COOH)₂;

—

N(CH₂COOH)₂;

—

SO₃H

—

OSO₃H

—

OPO₃H₂

—

PO₃H₂ or

-tetrazol-5-yl

and any Zn²⁺ complex thereof.

13. The pharmaceutical composition according to claim 1, wherein the parent insulin is a desB30 human insulin analogue.

14. The pharmaceutical composition according to claim 1 further comprising a rapid acting insulin

15. The pharmaceutical composition according to claim 1, wherein at least 85% of the rapid acting insulin is present as rapid acting insulin hexamer or complexes with a smaller molecular weight than rapid acting insulin hexamers.

16. The pharmaceutical composition according to claim 1, wherein the rapid acting insulin is AspB28 human insulin, LysB3 GluB29 human insulin and/or LysB28 ProB29 human insulin.

32. A pharmaceutical composition for the treatment of diabetes in a patient in need of such treatment, comprising a therapeutically effective amount of a pharmaceutical composition according to claim 1 together with a pharmaceutically acceptable carrier.

33. A method of treating diabetes in a patient in need of such a treatment, comprising administering to the patient a therapeutically effective amount of a pharmaceutical composition according to claim 1 together with a pharmaceutically acceptable carrier.

34. A method according to claim 33 for pulmonary treatment of diabetes.

17. A method for producing a pharmaceutical composition comprising an acylated insulin wherein more than about 4 zinc atoms per 6 molecules of acylated insulin are added to the composition.

18. A method according to claim 17 wherein up to about 12 zinc atoms per 6 molecules of acylated insulin are added to the composition.

19. A method according to claim 17 wherein between about 4.3 and about 12 zinc atoms per 6 molecules of acylated insulin are added to the composition.

20. A method according to claim 17 wherein the zinc is added to the composition before addition of a preservative.

21. A method according to claim 17 wherein the zinc is added to the composition after addition of a preservative.

22. A method according to claim 17, wherein part of the zinc is added before addition of a preservative and part of the zinc is added after addition of a preservative

23. A method according to claim 17, wherein the preservative is phenol and/or m-cresol.

24. A method according to claim 17, wherein a surfactant is mixed with the pharmaceutical composition.

25. A method according to claim 17, wherein acylated insulin has a side chain attached to the ε

-amino group of a Lys residue present in the B chain of the parent insulin, the side chain being of the general formula;

—

W—

X—

Y-Z₂

wherein W is;

an α

-amino acid residue having a carboxylic acid group in the side chain which residue forms, with one of its carboxylic acid groups, an amide group together with ε

-amino group of a Lys residue present in the B chain of the parent insulin;

a chain composed of two, three or four α

-amino acid residues linked together via amide carbonyl bonds, which chain—

via an amide bond—

is linked to an ε

-amino group of a Lys residue present in the B chain of the parent insulin, the amino acid residues of W being selected from the group of amino acid residues having a neutral side chain and amino acid residues having a carboxylic acid group in the side chain so that W has at least one amino acid residue which has a carboxylic acid group in the side chain;

or

a covalent bond from X to an ε

-amino group of a Lys residue present in the B chain of the parent insulin;

X is;

—

CO—

;

—

CH(COOH)CO—

;

—

CO—

N(CH₂COOH)CH₂CO—

;

—

CO—

N(CH₂COOH)CH₂CON(CH₂COOH)CH₂CO—

;

—

CO—

N(CH₂CH₂COOH)CH₂CH₂CO—

;

—

CO—

N(CH₂CH₂COOH)CH₂CH₂CON(CH₂CH₂COOH)CH₂CH₂CO—

;

—

CO—

NHCH(COOH)(CH₂)₄NHCO—

;

—

CO—

N(CH₂CH₂COOH)CH₂CO—

;

or

—

CO—

N(CH₂COOH)CH₂CH₂CO—

.

that

a) when W is an amino acid residue or a chain of amino acid residues, via a bond from the underscored carbon forms an amide bond with an amino group in W, or

b) when W is a covalent bond, via a bond from the underscored carbonyl carbon forms an amide bond with an ε

-amino group of a Lys residue present in the B chain of the parent insulin;

Y is;

—

(CH₂)_m—

where m is an integer in the range of 6 to 32;

a divalent hydrocarbon chain comprising 1, 2 or 3 —

CH═

CH—

groups and a number of —

CH₂—

groups sufficient to give a total number of carbon atoms in the chain in the range of 10 to 32; and

Z₂ is;

—

COOH;

—

CO-Asp;

—

CO-Glu;

—

CO-Gly;

—

CO-Sar;

—

CH(COOH)₂;

—

N(CH₂COOH)₂;

—

SO₃H;

or

—

PO₃H

and any Zn²⁺ complexes thereof, provided that when W is a covalent bond and X is —

CO—

, then Z is different from —

COOH.

26. A method according to claim 17, wherein the acylated insulin is having a formula

wherein Ins is the parent insulin moiety which via the α

-amino group of the N-terminal amino acid residue of the B chain or an ε

-amino group of a Lys residue present in the B chain of the insulin moiety is bound to the CO—

group in the side chain via an amide bond;

X₄ is

—

(CH₂)_n where n is 1, 2, 3, 4, 5 or 6;

NR, where R is hydrogen or —

(CH₂)_p—

COOH;

—

(CH₂)_p—

SO₃H;

—

(CH₂)_p—

PO₃H₂;

—

(CH₂)_p—

O—

SO₃H₂;

—

(CH₂)_p—

O—

PO₃H₂;

arylene substituted with 1 or 2 —

(CH₂)_p—

O—

COOH groups;

—

(CH₂)_p-tetrazolyl, where p is an integer in the range of 1 to 6;

—

(CR₁R₂)_q—

NR—

CO—

, where R₁ and R₄ independently of each other and independently for each value of q can be H, —

COOH, or OH, q is 1-6 and R is defined as above;

—

((CR₃R₄)_q1—

NR—

CO)_2-4—

, where R₃ and R₄ independently of each other and independently for each value of q₁ can be H, —

COOH, or OH, q₁ is 1-6 and R is defined as above;

or

a bond

W₁ is arylene or heteroarylene, which may be substituted with one or two groups selected from the group consisting of —

COOH, —

SO₃H, and —

PO₃H₂ and tetrazolyl, or W₁ is a bond;

m is 0, 1, 2, 3, 4, 5 or 6;

X₅ is

where R is defined as above;

or

a bond;

Y₁ is

—

(CR₁R₂)_q—

NR—

CO—

, where R₁ and R₂ independently of each other and independently for each value of q can be H, —

COOH, a bond or OH, q is 1-6; and

R is defined as above;

NR where R is defined as above;

—

((CR₃R₄)_q1—

NR—

CO)_2-4—

, where R₃ and R₄ independently of each other and independently for each value of q₁ can be H, —

COOH, or OH, q₁ is 1-6 and R is defined as above;

or

a bond;

Q₇ is

—

(CH₂)_r—

where r is an integer from 4 to 22;

a divalent hydrocarbon chain comprising 1, 2 or 3 —

CH═

CH—

groups and a number of —

CH₂—

groups sufficient to give a total number of carbon atoms in the chain in the range of 4 to 22;

or

a divalent hydrocarbon chain of the formula

—

(CH₂)_s-Q₈-(C₆H₄)_v1-Q₉-(CH₂)_W-Q₁₀-(C₆H₄)_v2-Q₁₁-(CH₂)_t-Q₁₂-(C₆H₄)_v3-Q₁₃-(CH₂)_z—

wherein Q₈-Q₁₃ independently of each other can be O;

S or a bond;

where s, w, t and z independently of each other are zero or an integer from 1 to 10 so that the sum of s, w, t and z is in the range from 4 to 22, and v₁, v₂, and v₃ independently of each other can be zero or 1, provided that when W₁ is a bond then Q₇ is not a divalent hydrocarbon chain of the formula —

(CH₂)_v4C₆H₄(CH₂)_W1—

wherein v₄ and w₁ are integers or one of them is zero so that the sum of v₄ and w1 is in the range of 6 to 22; and

Z₁ is;

—

COOH;

—

CO-Asp;

—

CO-Glu;

—

CO-Gly;

—

CO-Sar;

—

CH(COOH)₂;

—

N(CH₂COOH)₂;

—

SO₃H

—

PO₃H₂;

O—

SO₃H;

O—

PO₃H₂;

-tetrazolyl or

—

O—

W₂,

where W₂ is arylene or heteroarylene substituted with one or two groups selected from —

COOH, —

SO₃H, and —

PO₃H₂ and tetrazolyl;

provided that if W₁ is a bond and v₁, v₂ and v₃ are all zero and Q₈-13 are all a bonds, then Z₁ is O—

W₂

and any Zn²⁺ complex thereof.

27. A method according to claim 17, wherein the acylated insulin is having a formula

wherein Ins is the parent insulin moiety which via the α

-amino group of the N-terminal amino acid residue of the B chain or an ε

-amino group of a Lys residue present in the B chain of the insulin moiety is bound to the CO—

group in the side chain via an amide bond;

each n is independently 0, 1, 2, 3, 4, 5 or 6;

Q₁, Q₂, Q₃, and Q₄ independently of each other can be

(CH₂CH₂O)_s—

;

(CH₂CH₂CH₂O)_s—

;

(CH₂CH₂CH₂CH₂O)_s—

;

(CH₂CH₂OCH₂CH₂CH₂CH₂O)_s—

or (CH₂CH₂CH₂OCH₂CH₂CH₂CH₂O)_s—

where s is 1-20

—

(CH₂)_r—

where r is an integer from 4 to 22;

or a divalent hydrocarbon chain comprising 1, 2 or 3 —

CH═

CH—

groups and a number of —

CH₂—

groups sufficient to give a total number of carbon atoms in the chain in the range of 4 to 22;

—

(CH₂)_t—

or —

(CH₂OCH₂)_t—

, where t is an integer from 1 to 6;

—

(CR₁R₂)_q—

, where R₁ and R₂ independently of each other can be H, —

COOH, (CH₂)₁₆COOH and R₁ and R₂ can be different at each carbon, and q is 1-6,

—

((CR₃R₄)_q1)₁—

(NHCO—

(CR₃R₄)_q1—

NHCO)_1-2—

((CR₃R₄)_q1)₁ or —

((CR₃R₄)_q1)₁—

(CONH—

(CR₃R₄)_q1—

CONH)_1-2—

((CR₃R₄)_q1—

)—

, —

((CR₃R₄)_q1)₁—

(NHCO—

(CR₃R₄)_q1—

CONH)_1-2—

((CR₃R₄)_q1)₁ or —

((CR₃R₄)_q1)₁—

(CONH—

(CR₃R₄)_q1—

NHCO)_1-2—

((CR₃R₄)_q1)₁ where R₃ and R₄ independently of each other can be H, —

COOH, and R₃ and R₄ can be different at each carbon, and q1 is 1-6-, or

a bond;

with the proviso that Q₁-Q₄ are different;

X₁, X₂ and X₃ are independently

O;

a bond;

or

where R is hydrogen or —

(CH₂)_p—

COOH, —

(CH₂)_p—

SO₃H, —

(CH₂)_p—

PO₃H₂, —

(CH₂)_p—

O—

SO₃H;

—

(CH₂)_p—

O—

PO₃H₂;

or —

(CH₂)_p-tetrazol-5-yl, where each p independently of the other p'"'"'s is an integer in the range of 1 to 6; and

Z is;

—

COOH;

—

CO-Asp;

—

CO-Glu;

—

CO-Gly;

—

CO-Sar;

—

CH(COOH)₂;

—

N(CH₂COOH)₂;

—

SO₃H

—

OSO₃H

—

OPO₃H₂

—

PO₃H₂ or

-tetrazol-5-yl

and any Zn²⁺ complex thereof.

28. A method according to claim 17, wherein the parent insulin is a desB30 human insulin analogue.

29. A method according to claim 17, wherein a rapid acting insulin is mixed with the composition.

30. A method according to claim 29, wherein the rapid acting insulin is AspB28 human insulin, LysB3 GluB29 human insulin and/or LysB28 ProB29 human insulin.

31. (canceled)

35. (canceled)

36. (canceled)