US20090074882A1 - Insulin compositions and method of making a composition - Google Patents

Insulin compositions and method of making a composition Download PDF

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US20090074882A1
US20090074882A1 US12/096,476 US9647606A US2009074882A1 US 20090074882 A1 US20090074882 A1 US 20090074882A1 US 9647606 A US9647606 A US 9647606A US 2009074882 A1 US2009074882 A1 US 2009074882A1
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insulin
cooh
chain
ethoxy
bond
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Svend Havelund
Frantisek Hubalek
Helle Birk Olsen
Ib Jonassen
Thomas Hoeg-Jensen
Anne Plum
Ulla Ribel-Madsen
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Novo Nordisk AS
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Novo Nordisk AS
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/52—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an inorganic compound, e.g. an inorganic ion that is complexed with the active ingredient
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/28—Insulins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575—Hormones
    • C07K14/62—Insulins

Definitions

  • the present invention relates to pharmaceutical compositions of acylated insulin with a prolonged action profile and high zinc content. Further the invention relates to a method for producing a composition with a prolonged action profile and high zinc content and a method for manufacturing a composition for the treatment of diabetes.
  • the treatment of diabetes both type 1 diabetes and type 2 diabetes, relies to an increasing extent on the so-called intensive insulin treatment.
  • the patients are treated with multiple daily insulin injections comprising one or two daily injections of a long acting insulin to cover the basal insulin requirement supplemented by bolus injections of a rapid acting insulin to cover the insulin requirement related to meals.
  • Long acting insulin compositions are well known in the art.
  • one main type of long acting insulin compositions comprises injectable aqueous suspensions of insulin crystals or amorphous insulin.
  • the insulin compounds utilized typically are protamine insulin, zinc insulin or protamine zinc insulin.
  • protamine insulin crystals can be immunogenic in man and that their use for medical purposes may lead to formation of antibodies. Also, evidence has been found that the protamine crystal is itself immunogenic. Therefore, with some patients the use of long acting insulin compositions containing protamines must be avoided.
  • Another type of long acting insulin compositions are solutions having a pH value below physiological pH from which the insulin will precipitate because of the rise in the pH value when the solution is injected.
  • a drawback with these solutions is that the particle size distribution of the precipitate formed in the tissue on injection, and thus the release profile of the medication, depends on the blood flow at the injection site and other parameters in a somewhat unpredictable manner.
  • a further drawback is that the solid particles of the insulin may act as a local irritant causing inflammation of the tissue at the site of injection.
  • Insulin is a 51 amino acid peptide hormone produced in the islets of Langerhans in the pancreas. Its primary function, acting as a monomer, is to facilitate the transport of glucose molecules across the cell membranes of adipose and muscle tissue by binding to and activating a transmembrane receptor.
  • a distinctive property of insulin is its ability to associate into hexamers, in which form the hormone is protected from chemical and physical degradation during biosynthesis and storage.
  • X-ray crystallographic studies on insulin show that the hexamer consists of three dimers related by a 3-fold axis of rotation. These dimers are closely associated through the interaction of two zinc ions at its core positioned on the 3-fold axis.
  • Zinc and phenolic additives are regularly used in therapeutic insulin preparations to promote hexamer formation as a precaution against degradation during storage. In this form, however, the action of injected insulin is delayed while the hexamers diffuse through the subcutis and dissociate into dimers and monomers.
  • Formulations of insulin are usually prepared by dissolving insulin in a small volume of water under acidic conditions. Zinc is then added to the formulation followed by a neutralisation and addition of preservatives like phenol and m-cresol.
  • the pharmaceutical formulation of these insulins are given as about 2, 3 or 4 zinc atoms per hexamer insulin.
  • WO 2005/012347 discloses another group of acylated insulin derivatives comprising additional negatively charge compared to the acylated insulins disclosed in WO 95/07931.
  • the pharmaceutical formulation of these acylated insulins are given as 2, 3 or 4 zinc atoms per hexamer insulin.
  • WO 2003/094956 disclose stable insulin formulations prepared by mixing a monomeric insulin and a soluble acylated insulin analogue.
  • the formulations contains from about 2.3 to about 4.5 Zn 2+ per hexamer insulin.
  • the acylated insulin analogue according to this invention is insulin detemir.
  • acylated insulin analogue is insulin detemir.
  • WO 99/21888 concerns aggregates of human insulin derivatives, which contains up to 5 zinc atoms per 6 molecules of insulin derivate.
  • Human insulin has three primary amino groups: the N-terminal group of the A-chain and of the B-chain and the ⁇ -amino group of Lys B29 .
  • Several insulin derivatives which are substituted in one or more of these groups are known in the prior art.
  • U.S. Pat. No. 3,528,960 (Eli Lilly) relates to N-carboxyaroyl insulins in which one, two or three primary amino groups of the insulin molecule has a carboxyaroyl group.
  • EP 894095 discloses insulin derivatives wherein the N-terminal group of the B-chain and/or the ⁇ -amino group of Lys in position B28, B29 or B30 has a substituent of the formula —CO—W—COOH where W can be a long chain hydrocarbon group. These insulin derivatives have a prolonged profile of action and are soluble at physiological pH values.
  • WO 95/07931 discloses protracted insulin derivatives wherein a liphophilic side chain is attached either to the ⁇ -amino group of the N-terminal amino acid residue of the B chain or to the ⁇ -amino group of a Lys residue present in the B chain of the parent insulin.
  • acylated insulin analogue is prepared.
  • the insulin formulation is prepared by adding 2-2.5 zinc atoms per hexamer insulin followed by glycerol and phenol.
  • the structures of the insulin is measured by size exclusion chromatography.
  • the present invention is related to certain pharmaceutical compositions of acylated insulins which solves the problems of the prior art.
  • the present invention is related to a soluble pharmaceutical composition
  • a soluble pharmaceutical composition comprising an acylated insulin comprising more than 4 zinc atoms per 6 molecules of the acylated insulin.
  • the zinc content may be up to about 12 zinc atoms per 6 molecules of acylated insulin.
  • the upper limit for the zinc content is the content of zinc which would cause precipitation of the insulin and turning the solution into a suspension.
  • the pharmaceutical composition comprises between about 4.3 and about 12 zinc atoms per 6 molecules of acylated insulin or between about 4.5 and about 12 zinc atoms per 6 molecules of acylated insulin. In a further aspect of the invention the pharmaceutical composition comprises between about 5 and about 11.4 zinc atoms per 6 molecules of acylated insulin or between about 5.5 and about 10 zinc atoms per 6 molecules of acylated insulin. In a further aspect the pharmaceutical composition comprises between about 6 and about 10.5 zinc atoms per 6 molecules of acylated insulin or the pharmaceutical composition comprises between about 6.5 and about 10 zinc atoms per 6 molecules of acylated insulin or the pharmaceutical composition comprises between about 7 and about 9 zinc atoms per 6 molecules of acylated insulin. In a further aspect the pharmaceutical composition comprises citrate from about one third to about 3 times the zinc concentration.
  • the pharmaceutical composition comprises a surfactant.
  • the surfactant can be present in an amount of 0.0005-0.01% based on the weight of the pharmaceutical composition.
  • the surfactant can be present in an amount of 0.0005-0.007% based on the weight of the composition.
  • An example of a surfactant could be polysorbate 20, which can be present in the composition in an amount of 0.001-0.003% based on the weight of the composition.
  • Another example is poloxamer 188, which can be present in an amount of 0.002-0.006% based on the weight of the composition.
  • the insulin molecule may be acylated at various positions in the insulin molecule.
  • the insulin is acylated in the F-amino group of a Lys residue in a position in the B-chain of the parent insulin molecule in particularly in the ⁇ -amino group of the B29 lysine group in the human insulin molecule.
  • the acylation may take place in another position in the insulin molecule, e.g. the ⁇ -amino group in position B1 or in position where the natural amino acid residue in the insulin molecule has been substituted with a lysine residue provided that B29 is changed from a lysine to another amino acid residue.
  • the acylated insulin is acylated either in the ⁇ -amino group in the B1 position or in a free ⁇ -amino group of a lysine residue in the A- or B-chain of the insulin molecule.
  • the insulin is acylated in the free ⁇ -amino group of the lysine residue in position B29 of the insulin molecule.
  • the acyl group will be a liphophilic group and will typically be a fatty acid moiety having from about 6 to about 32 carbon atoms comprising at least one free carboxylic acid group or a group which is negatively charged at neutral pH.
  • the fatty acid moiety will more typically have from 6 to 24, from 8 to 20, from 12 to 20, from 12-16, from 10-16, from 10-20, from 14-18 or from 14-16 carbon atoms.
  • the fatty acid moiety is derived from a dicarboxylic fatty acid with from about 6 to about 32, from 6 to 24, from 8 to 20, from 12 to 20, from 12-16, from 10-16, from 10-20, from 14-18 or from 14-16 carbon atoms.
  • the pharmaceutical composition comprises an acyl group which is attached to the insulin via a linker group through amide bonds.
  • the acyl group may be attached directly to the free amino group in question. However, the acyl group may also be attached via amide bonds by a linker which links the free amino group in the insulin molecule and the acyl group in question together.
  • the pharmaceutical composition comprises an insulin wherein the insulin has a side chain attached either to the ⁇ -amino group of the N-terminal amino acid residue of the B chain or to an ⁇ -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.
  • the insulin molecule has a side chain attached to the ⁇ -amino group of a Lys residue present in the B chain of the parent insulin, the side chain being of the general formula:
  • W is:
  • X is:
  • Y is:
  • the B30 amino acid residue has been deleted and the acylated insulin is a desB30 insulin.
  • W is an ⁇ -amino acid residue having from 4 to 10 carbon atoms and in a further aspect W is selected from the group consisting of ⁇ -Asp, ⁇ -Asp, ⁇ -Glu, ⁇ -Glu, ⁇ -hGlu and ⁇ -hGlu.
  • X is —CO—.
  • Z 2 is —COOH
  • the substructure Y of the side chain —W—X—Y— Z 2 can be a group of the formula —(CH 2 ) m — where m is an integer in the range of from 6 to 32, from 8 to 20, from 12 to 20, or from 12-16.
  • Y is a divalent hydrocarbon chain comprising 1, 2 or 3 —CH ⁇ CH— groups and a number of —CH 2 — groups sufficient to give a total number of carbon atoms in the chain in the range of from 6 to 32, from 10 to 32, from 12 to 20, or from 12-16.
  • W is selected from the group consisting of ⁇ -Asp, ⁇ -Asp, ⁇ -Glu, and ⁇ -Glu;
  • X is —CO— or —CH(COOH)CO;
  • Y is —(CH 2 ) m — where m is an integer in the range of 12-18 and Z 2 is —COOH or —CH(COOH) 2 .
  • Non limiting examples of acylated insulin compounds are N ⁇ B29 —(N ⁇ -(HOOC(CH 2 ) 14 CO)- ⁇ -Glu) desB30 human insulin; N ⁇ B29 —(N ⁇ -(HOOC(CH 2 ) 15 CO)- ⁇ -Glu) desB30 human insulin; N ⁇ B29 —(N ⁇ -(HOOC(CH 2 ) 16 CO)- ⁇ -Glu) desB30 human insulin; N ⁇ B29 -(N ⁇ -(HOOC(CH 2 ) 17 CO)- ⁇ -Glu) desB30 human insulin; N ⁇ B29 —(N ⁇ -(HOOC(CH 2 ) 18 CO)- ⁇ -Glu) desB30 human insulin; N ⁇ B29 —(N ⁇ -(HOOC(CH 2 ) 16 CO)- ⁇ -Glu-N-( ⁇ -Glu)) desB30 human insulin; N ⁇ B29 —(N ⁇ -(Asp-OC(CH 2 )
  • the side chain may comprise at least one aromatic group or at least one difunctional PEG group.
  • PEG polyethyleneglycol
  • the acylated insulin used in the pharmaceutical composition is having a formula
  • Ins is the parent insulin moiety which via the ⁇ -amino group of the N-terminal amino acid residue of the B chain or an ⁇ -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;
  • Q 8 -Q 13 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 v 1 , v 2 , and v 3 independently of each other can be zero or 1, provided that when W 1 is a bond then Q 7 is not a divalent hydrocarbon chain of the formula —(CH 2 ) v4 C 6 H 4 (CH 2 ) W1 — wherein v 4 and w 1 are integers or one of them is zero so that the sum of v 4 and w 1 is in the range of 6 to 22; and
  • W 1 is phenylene. In one aspect W 1 is 5-7 membered heterocyclic ring system comprising nitrogen, oxygen or sulphur. In one aspect W 1 is a 5 membered heterocyclic ring system comprising at least one oxygen.
  • Q 7 is —(CH 2 ) r — where r is an integer in the range of from 4 to 22, from 8- to 20, from 12 to 20 or from 14-18.
  • Q 8 , Q 9 , Q 12 and Q 13 are all bonds, v 2 is 1 and v 1 and v 3 are zero.
  • Q 10 and Q 11 are oxygen.
  • X 4 and Y 1 are a bonds and X 5 is
  • R is —(CH 2 ) p —COOH, where p is 1-4.
  • Z 1 is —COOH
  • acylated insulin of the pharmaceutical composition is selected from the group consisting of
  • the acylated insulin present in the pharmaceutical composition is having a formula
  • Ins is the parent insulin moiety which via the ⁇ -amino group of the N-terminal amino acid residue of the B chain or an ⁇ -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; Q 1 , Q 2 , Q 3 , and Q 4 independently of each other can be
  • R is hydrogen or —(CH 2 ) p —COOH, —(CH 2 ) p —SO 3 H, —(CH 2 ) p —PO 3 H 2 , —(CH 2 ) p —O—SO 3 H; —(CH 2 ) p —O—PO 3 H 2 ; or —(CH 2 ) 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
  • the acylated insulin of the pharmaceutical composition is selected from the group consisting of N ⁇ B29 -(3- ⁇ 2-[2-(2-[ ⁇ -carboxy-pentadecanoyl- ⁇ -glutamyl-(2-amino-ethoxy)]-ethoxy)-ethoxy ⁇ -ethoxy]-propinoyl) desB30 human insulin, N ⁇ B29 -(3- ⁇ 2-[2-(2-[ ⁇ -carboxy-heptadecanoyl- ⁇ -glutamyl-(2-amino-ethoxy)]-ethoxy)-ethoxy ⁇ -ethoxy]-propinoyl) desB30 human insulin, N ⁇ B29 - ⁇ 3-[2-(2- ⁇ 2-[2-( ⁇ -carboxy-pentadecanoylamino)ethoxy]-ethoxy ⁇ -ethoxy)-ethoxy]-propionyl- ⁇ -glutamyl desB30 human insulin, N
  • the parent insulin molecule is human insulin or an analogue thereof.
  • Non limiting analogues of human insulin is desB30 analogue; insulin analogues where the amino acid residue in position B30 is Lys and the amino acid residue in position B29 is any codable amino acid except Cys, Met, Arg and Lys; insulin analogues where the amino acid residue at position A21 is Asn and insulin analogues where the amino acid residue at position B3 is Lys and the amino acid residue at position B29 is Glu.
  • amino acid residue at position B28 is Asp.
  • a specific example from this group of parent insulin analogues is AspB28 human insulin disclosed in EP 214826.
  • the amino acid residue at position B28 is Lys and the amino acid residue at position B29 is Pro.
  • a specific example from this group of parent insulin analogues is Lys B28 Pro B29 human insulin.
  • the amino acid residue in position B30 is Lys and the amino acid residue in position B29 is any codable amino acid except Cys, Met, Arg and Lys.
  • An example is an insulin analogue where the amino acid residue at position B29 is Thr and the amino acid residue at position B30 is Lys.
  • a specific example from this group of parent insulin analogues is Thr B29 Lys B30 human insulin.
  • the amino acid residue at position B3 is Lys and the amino acid residue at position B29 is Glu.
  • a specific example from this group of parent insulin analogues is Lys B3 Glu B29 human insulin.
  • the pharmaceutical composition according to the present invention will comprise a therapeutically effective amount of the acylated insulin together with a pharmaceutically acceptable carrier for the treatment of type 1 diabetes, type 2 diabetes and other states that cause hyperglycaemia in patients in need of such a treatment.
  • a pharmaceutical composition for treating type 1 diabetes, type 2 diabetes and other states that cause hyperglycaemia in a patient in need of such a treatment comprising a therapeutically effective amount of an acylated insulin derivative as defined above in mixture with an insulin or an insulin analogue which has a rapid onset of action, together with pharmaceutically acceptable carriers and additives.
  • the pharmaceutical composition may comprise a mixture of two insulin components: one with a protracted insulin action, a basal insulin, and the other with a rapid onset of action, a bolus insulin.
  • An example of such mixture is Insulin aspart, AspB28 human insulin in mixture with N ⁇ B29 —(N ⁇ -(HOOC(CH 2 ) 14 CO)- ⁇ -Glu) desB30 human insulin corresponding to LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin disclosed in WO 2005/012347.
  • Lispro Lys B28 Pro B29 human insulin, in mixture with LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin.
  • a third example of such a mixture is Glulisine, Lys B3 Glu B29 -human insulin, in mixture with LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin.
  • 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.
  • At least 90, 92, 95, 96, 97, 98, 99, 99.5% of the rapid acting insulin is present as rapid acting insulin hexamer or complexes with a smaller molecular weight than rapid acting insulin hexamers.
  • the acylated insulin derivative and the rapid acting insulin analogue can be mixed in a molar ratio about 90%/10%; about 75%/25%, about 70%/30% about 50%/50%, about 25%/75%, about 30%/70% or about 10%/90%.
  • the pharmaceutical composition according to the invention will have a pH between about 6.5 to about 8.5.
  • the pH is from about 7.0 to about 8.2
  • the pH is from about 7.2 to 8.0 or from about 7.4 to about 8.0 or the pH is from about 7.4 to about 7.8.
  • the invention further comprises 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.
  • more than about 4.3 zinc atoms per 6 molecules of acylated insulin are added to the composition or more than about 4.5 zinc atoms per 6 molecules of acylated insulin are added to the composition or than about 5 zinc atoms per 6 molecules of acylated insulin are added to the composition.
  • more than about 5.5 zinc atoms or more than about 6.5 zinc atoms, or more than about 7.0 zinc atoms or more than about 7.5 zinc atoms per 6 molecules of acylated insulin are added to the composition.
  • the method comprises adding up to about 12 zinc atoms per 6 molecules of acylated insulin to the composition. In one aspect of the invention the method comprises adding between about 4.3 and about 12 zinc atoms per 6 molecules of acylated insulin to the composition
  • zinc atoms per 6 molecules of acylated insulin are added to the composition or about 5 and about 11.4 zinc atoms per 6 molecules of acylated insulin are added to the composition or between about 5.5 and about 10 zinc atoms per 6 molecules of acylated insulin are added to the composition.
  • the method comprises adding zinc to the composition before the addition of a preservative.
  • the number of zinc atoms added before addition of a preservative is more than 1 zinc atom per 6 molecules of acylated insulin, or the number of zinc atoms added before addition of a preservative is more than 2 zinc atom per 6 molecules of acylated insulin or the number of zinc atoms added before addition of a preservative is more than 3 zinc atom per 6 molecules of acylated insulin or the number of zinc atoms added before addition of a preservative is more than 4 zinc atom per 6 molecules of acylated insulin or the number of zinc atoms added before addition of a preservative is more than 5 zinc atom per 6 molecules of acylated insulin.
  • zinc atoms per 6 molecules of acylated insulin are added to the composition before the addition of a preservative or about 5 and about 11.4 zinc atoms per 6 molecules of acylated insulin are added to the composition before the addition of a preservative or even more preferred between about 5.5 and about 10 zinc atoms per 6 molecules of acylated insulin are added to the composition before the addition of a preservative.
  • the method comprises adding zinc to the composition after addition of a preservative.
  • at least 0.5 zinc atom per 6 molecules of acylated insulin is added to the composition after addition of a preservative or at least 1 zinc atom per 6 molecules of acylated insulin is added to the composition after addition of a preservative.
  • more than about 2 zinc atoms per 6 molecules of acylated insulin are added to the composition after the addition of a preservative or more than about 3 zinc atoms per 6 molecules of acylated insulin are added to the composition after the addition of a preservative or more than about 4 zinc atoms per 6 molecules of acylated insulin are added to the composition after the addition of a preservative.
  • between about 0.5 and about 12, between about 1 and about 11.4, between about 1.5 and about 11, between about 2 and about 10.5, between about 3 and about 10 or between about 4 and about 9 zinc atoms per 6 molecules of acylated insulin are added to the composition after the addition of a preservative.
  • zinc atoms per 6 molecules of acylated insulin are added to the composition after the addition of a preservative or about 5 and about 11.4 zinc atoms per 6 molecules of acylated insulin are added to the composition after the addition of a preservative or between about 5.5 and about 10 zinc atoms per 6 molecules of acylated insulin are added to the composition after the addition of a preservative.
  • the method comprises adding part of the zinc before addition of a preservative and adding part of the zinc after addition of a preservative.
  • the method comprises adding at least 0.5 zinc atom per 6 molecules of acylated insulin before addition of a preservative and adding at least 1 zinc atom per 6 molecules of acylated insulin after addition of a preservative. In one aspect the method comprises at least 0.5 zinc atom per 6 molecules of acylated insulin before addition of a preservative and adding at least 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 zinc atoms per 6 molecules of acylated insulin after addition of a preservative.
  • the method comprises adding at least 1 zinc atom per 6 molecules of acylated insulin before addition of a preservative and adding at least 1 zinc atom per 6 molecules of acylated insulin after addition of a preservative or adding at least 1 zinc atom per 6 molecules of acylated insulin before addition of a preservative and adding at least 2 or 3 zinc atoms per 6 molecules of acylated insulin after addition of a preservative or adding at least 1 zinc atom per 6 molecules of acylated insulin before addition of a preservative and up to about 11 zinc atom per 6 molecules of acylated insulin after addition of a preservative.
  • the method comprises adding at least 1 zinc atom per 6 molecules of acylated insulin before addition of a preservative and adding at least 4, 5, 6, 7, 8, 9, 10 or 11 zinc atoms per 6 molecules of acylated insulin after addition of a preservative.
  • the method comprises adding at least 2 zinc atoms per 6 molecules of acylated insulin before addition of a preservative and adding at least 1 zinc atom per 6 molecules of acylated insulin after addition of a preservative or adding at least 2 zinc atoms per 6 molecules of acylated insulin before addition of a preservative and adding at least 2 or 3 zinc atoms per 6 molecules of acylated insulin after addition of a preservative or adding at least 2 zinc atoms per 6 molecules of acylated insulin before addition of a preservative and up to about 10 zinc atoms per 6 molecules of acylated insulin after addition of a preservative
  • the method comprises adding at least 2 zinc atom per 6 molecules of acylated insulin before addition of a preservative and adding at least 4, 5, 6, 7, 8, 9 or 10 zinc atoms per 6 molecules of acylated insulin after addition of a preservative.
  • the method comprises adding at least 3 zinc atoms per 6 molecules of acylated insulin before addition of a preservative and adding at least 1 zinc atom per 6 molecules of acylated insulin after addition of a preservative or adding at least 3 zinc atoms per 6 molecules of acylated insulin before addition of a preservative and adding at least 2 or 3 zinc atoms per 6 molecules of acylated insulin after addition of a preservative or adding at least 3 zinc atoms per 6 molecules of acylated insulin before addition of a preservative and up to about 9 zinc atoms per 6 molecules of acylated insulin after addition of a preservative
  • the method comprises adding the whole amount of zinc atoms to the pharmaceutical composition in one step.
  • the method comprises adding the zinc atoms to the pharmaceutical composition in two or more steps.
  • zinc may be added to the composition in one, two, three, four or five steps, where each step includes addition of small amounts of max. 1 Zn/6ins.
  • the zinc may be added to the composition in one, two, three, four or five steps, where each step includes addition of small amounts of 2 Zn/6ins, 3 Zn/6ins, 4 Zn/6ins, 5 Zn/6ins or 6 Zn/6ins.
  • the method comprises adding a surfactant to the pharmaceutical composition.
  • the surfactant can be mixed in the pharmaceutical composition in an amount of 0.0005-0.01% based on the weight of the pharmaceutical composition.
  • the surfactant can be mixed in the pharmaceutical composition in an amount of 0.0005-0.007% based on the weight of the composition.
  • An example of a surfactant could be polysorbate 20, which can be mixed in the pharmaceutical composition in an amount of 0.001-0.003% based on the weight of the composition.
  • poloxamer 188 which can be mixed in the pharmaceutical composition in an amount of 0.002-0.006% based on the weight of the composition.
  • Ins is the parent insulin moiety which via the ⁇ -amino group of the N-terminal amino acid residue of the B chain or an ⁇ -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;
  • W 1 is phenylene. In one aspect W 1 is 5-7 membered heterocyclic ring system comprising nitrogen, oxygen or sulphur. In one aspect W 1 is a 5 membered heterocyclic ring system comprising at least one oxygen.
  • R is —(CH 2 ) p —COOH, where p is 1-4.
  • Z 1 is —COOH
  • Ins is the parent insulin moiety which via the ⁇ -amino group of the N-terminal amino acid residue of the B chain or an ⁇ -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; Q 1 , Q 2 , Q 3 , and Q 4 independently of each other can be
  • Z is —COOH
  • the acylated insulin used in the method for preparing a pharmaceutical composition is selected from the group consisting of N ⁇ B29 -(3-[2- ⁇ 2-(2-[ ⁇ -carboxy-pentadecanoyl- ⁇ -glutamyl-(2-amino-ethoxy)]-ethoxy)-ethoxy ⁇ -ethoxy]-propinoyl) desB30 human insulin, N ⁇ B29 -(3-[2- ⁇ 2-(2-[ ⁇ -carboxy-heptadecanoyl- ⁇ -glutamyl-(2-aminoethoxy)]-ethoxy)-ethoxy ⁇ -ethoxy]-propinoyl) desB30 human insulin, N ⁇ B29 - ⁇ 3-[2-(2- ⁇ 2-[2-( ⁇ -carboxy-pentadecanoylamino)-ethoxy]-ethoxy ⁇ -ethoxy)-ethoxy]-propionyl- ⁇ -glutamyl des
  • the method comprises adding a rapid acting insulin to the composition.
  • the rapid acting insulin is AspB28human insulin, LysB28ProB29 human insulin and Lys B3 Glu B29 -human insulin or a mixture thereof.
  • the pharmaceutical composition comprising acylated insulin is used for treatment of diabetes.
  • the pharmaceutical composition comprising acylated insulin is used for the manufacture of a medicament for the treatment of diabetes.
  • a method of treating type 1 diabetes, type 2 diabetes and other states that cause hyperglycaemia in a patient in need of such a treatment comprising administering to the patient a therapeutically effective amount of an pharmaceutical composition together with a pharmaceutically acceptable carrier and/or pharmaceutical acceptable additives.
  • a method for the manufacture of a pharmaceutical composition for the use in the treatment of type 1 diabetes, type 2 diabetes and other states that cause hyperglycaemia is provided.
  • a method of treating type 1 diabetes, type 2 diabetes and other states that cause hyperglycaemia in a patient in need of such a treatment comprising administering to the patient a therapeutically effective amount of an pharmaceutical composition according to the invention.
  • a method for the manufacture of a pharmaceutical composition for the use in the treatment of type 1 diabetes, type 2 diabetes and other states that cause hyperglycaemia is provided.
  • the pharmaceutical composition comprising an acylated insulin as defined in the pre-sent specification may be administered simultaneously or sequentially with OAD(s) or GLP-1.
  • the factors may be supplied in single-dosage form wherein the single-dosage form contains both compounds, or in the form of a kit-of-parts comprising a preparation of a the pharmaceutical composition comprising a pharmaceutical composition comprising an acylated insulin and a pharmaceutical composition containing an OAD as a second unit dosage form.
  • a first or second or third, etc., unit dose is mentioned throughout this specification this does not indicate the preferred order of administration, but is merely done for convenience purposes.
  • a preparation of a pharmaceutical composition comprising an acylated insulin and a preparation of OAD(s) or GLP-1 is meant administration of the compounds in single-dosage form, or administration of a first agent followed by administration of a second agent with a time separation of no more than 15 minutes, 10, 5 or 2 minutes. Either factor may be administered first.
  • sequential dosing administration of a first agent followed by administration of a second agent with a time separation of more than 15 minutes.
  • Either of the two unit dosage form may be administered first.
  • both products are injected through the same intravenous access.
  • the pharmaceutical composition comprising an acylated insulin is administered once daily simultaneously or sequentially with OAD(s) or GLP-1.
  • the pharmaceutical composition comprising an acylated insulin further comprises a rapid acting insulin and is administered once daily together with OAD(s) or GLP-1.
  • the pharmaceutical composition comprising an acylated insulin can be a stand alone intensive treatment given up till 5 times daily.
  • the pharmaceutical composition further comprises a rapid acting insulin, where the intensive treatment can be a stand alone treatment given up till 5 times daily.
  • a soluble pharmaceutical composition comprising an acylated insulin and further comprising more than 4 zinc atoms per 6 molecules of acylated insulin.
  • composition according to paragraph 1 comprising up to about 12 zinc atoms per 6 molecules of acylated insulin.
  • composition according to paragraphs 1 or 2 comprising between about 4.3 and about 12 zinc atoms per 6 molecules of acylated insulin.
  • composition according to any of paragraphs 1-3 comprising between about 4.5 and about 12 zinc atoms per 6 molecules of acylated insulin.
  • composition according to any of the preceding paragraphs, 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.
  • composition according to any of the preceding paragraphs wherein at least 92% of the acylated insulin is present as complexes which are acylated insulin dodecamers or complexes with a higher molecular weight than acylated insulin dodecamer.
  • composition according to any of the preceding paragraphs wherein at least 95% of the acylated insulin is present as complexes which are acylated insulin dodecamers or complexes with a higher molecular weight than acylated insulin dodecamer.
  • composition according to any of the preceding paragraphs, wherein at least 97% of the acylated insulin is present as complexes which are acylated insulin dodecamers or complexes with a higher molecular weight than acylated insulin dodecamer.
  • composition according to any of the preceding paragraphs, wherein the composition comprises a surfactant.
  • acylated insulin is an insulin acylated in the ⁇ -amino group of a Lys residue in a position in the B-chain of the parent insulin molecule.
  • composition according to any of paragraphs 1-10, wherein the acyl group comprises at least one free carboxylic acid or a group which is negatively charged at neutral pH.
  • composition according to paragraph 1 or 10-11, wherein the acyl group is derived from a dicarboxylic fatty acid with from 4 to 32 carbon atoms.
  • composition according to paragraph 1 or 10-12 wherein the acyl group is attached to the insulin molecule via a linker group through amide bonds.
  • linker group comprises at least one free carboxylic group or a group which is negatively charged at neutral pH.
  • composition according to any of paragraphs 1-14 wherein the insulin molecule has a side chain attached either to the ⁇ -amino group of the N-terminal amino acid residue of the B chain or to an ⁇ -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.
  • composition according to paragraphs 1-15 wherein the side chain comprises at least one difunctional PEG group.
  • composition according to any of paragraphs 1-15 wherein the insulin molecule has a side chain attached to the ⁇ -amino group of a Lys residue present in the B chain of the parent insulin, the side chain being of the general formula:
  • W is:
  • X is:
  • Y is:
  • composition according to paragraphs 1-15 and 18-19 wherein the acylated insulin is selected from the group consisting of N ⁇ B29 —(N ⁇ -(HOOC(CH 2 ) 14 CO)- ⁇ -Glu) desB30 human insulin; N ⁇ B29 —(N—(HOOC(CH 2 ) 15 CO)- ⁇ -Glu) desB30 human insulin; N ⁇ B29 -(N—(HOOC(CH 2 ) 16 CO)- ⁇ -Glu) desB30 human insulin; N ⁇ B29 —(N ⁇ -(HOOC(CH 2 ) 17 CO)- ⁇ -Glu) desB30 human insulin; N ⁇ B29 —(N ⁇ -(HOOC(CH 2 ) 18 CO)- ⁇ -Glu) desB30 human insulin; N ⁇ B29 -(N—(HOOC(CH 2 ) 16 CO)- ⁇ -Glu-N-( ⁇ -Glu)) desB30 human insulin; N ⁇ B29 —(N ⁇ -(Asp
  • composition according to paragraph 1-16, wherein the acylated insulin is having a formula
  • Ins is the parent insulin moiety which via the ⁇ -amino group of the N-terminal amino acid residue of the B chain or an ⁇ -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;
  • composition according paragraph 21-24 wherein Q 8 , Q 9 , Q 12 and Q 13 are all bonds, v 2 is 1 and v 1 and v 3 are zero.
  • R is —(CH 2 ) p —COOH, where p is 1-4.
  • Ins is the parent insulin moiety which via the ⁇ -amino group of the N-terminal amino acid residue of the B chain or an ⁇ -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; Q 1 , Q 2 , Q 3 , and Q 4 independently of each other can be
  • composition according to any of the preceeding paragraphs, wherein the parent insulin is a desB30 human insulin analogue.
  • composition according to any of the preceding paragraphs, wherein the parent insulin is selected from the group consisting of human insulin; desB1 human insulin; desB30 human insulin; GlyA21 human insulin; GlyA21 desB30 human insulin; AspB28 human insulin; porcine insulin; LysB28 ProB29 human insulin; GlyA21 ArgB31 ArgB32 human insulin; and LysB3 GluB29 human insulin or AspB28 desB30 human insulin.
  • the parent insulin is selected from the group consisting of human insulin; desB1 human insulin; desB30 human insulin; GlyA21 human insulin; GlyA21 desB30 human insulin; AspB28 human insulin; porcine insulin; LysB28 ProB29 human insulin; GlyA21 ArgB31 ArgB32 human insulin; and LysB3 GluB29 human insulin or AspB28 desB30 human insulin.
  • composition according to paragraphs 1-16, 21-29 and 34-35, wherein the acylated insulin is selected from the group consisting of
  • composition according to paragraphs 1-15, 17 and 30-35 wherein the acylated insulin is selected from the group consisting of N ⁇ B29 -(3- ⁇ 2-[2-(2-[ ⁇ -carboxy-pentadecanoyl- ⁇ -glutamyl-(2-amino-ethoxy)]-ethoxy)-ethoxy ⁇ -ethoxy]-propinoyl) desB30 human insulin, N ⁇ B29 -(3- ⁇ 2-[2-(2-[ ⁇ -carboxy-heptadecanoyl- ⁇ -glutamyl-(2-aminoethoxy)]-ethoxy)-ethoxy ⁇ -ethoxy]-propinoyl) desB30 human insulin, N ⁇ B29 - ⁇ 3-[2-(2- ⁇ 2-[2-( ⁇ -carboxy-pentadecanoylamino)-ethoxy]-ethoxy ⁇ -ethoxy)-ethoxy]-propionyl- ⁇ -glutamyl
  • composition according to any of the preceeding paragraphs having a pH between about 6.5 and 8.5
  • composition according to any of the preceding paragraphs further comprising a rapid acting insulin
  • composition according to paragraphs 1 and 39 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.
  • composition according to paragraphs 39-44, wherein the rapid acting insulin is AspB28 human insulin, LysB3 GluB29 human insulin and/or LysB28 ProB29 human insulin.
  • 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.
  • acylated insulin has a side chain attached to the ⁇ -amino group of a Lys residue present in the B chain of the parent insulin, the side chain being of the general formula:
  • W is:
  • X is:
  • Y is:
  • acylated insulin is selected from the group consisting of N ⁇ B29 —(N—(HOOC(CH 2 ) 14 CO)- ⁇ -Glu) desB30 human insulin; N ⁇ B29 —(N ⁇ -(HOOC(CH 2 ) 15 CO)- ⁇ -Glu) desB30 human insulin; N ⁇ B29 —(N ⁇ -(HOOC(CH 2 ) 16 CO)- ⁇ -Glu) desB30 human insulin; N ⁇ B29 —(N ⁇ -(HOOC(CH 2 ) 17 CO)- ⁇ -Glu) desB30 human insulin; N ⁇ B29 —(N ⁇ -(HOOC(CH 2 ) 18 CO)- ⁇ -Glu) desB30 human insulin; N ⁇ B29 -(N—(HOOC(CH 2 ) 16 CO)- ⁇ -Glu-N-( ⁇ -Glu)) desB30 human insulin; N ⁇ B29 —(N ⁇ ⁇ -(HOOC(CH 2 ) 16 CO)- ⁇ -
  • Ins is the parent insulin moiety which via the ⁇ -amino group of the N-terminal amino acid residue of the B chain or an ⁇ -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;
  • Q 8 -Q 13 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 v 1 , v 2 , and v 3 independently of each other can be zero or 1, provided that when W 1 is a bond then Q 7 is not a divalent hydrocarbon chain of the formula —(CH 2 ) v4 C 6 H 4 (CH 2 ) W1 — wherein v 4 and w 1 are integers or one of them is zero so that the sum of v 4 and w 1 is in the range of 6 to 22; and
  • R is —(CH 2 ) p —COOH, where p is 1-4.
  • Ins is the parent insulin moiety which via the ⁇ -amino group of the N-terminal amino acid residue of the B chain or an ⁇ -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; Q 1 , Q 2 , Q 3 , and Q 4 independently of each other can be
  • parent insulin is selected from the group consisting of human insulin; desB1 human insulin; desB30 human insulin; GlyA21 human insulin; GlyA21 desB30 human insulin; AspB28 human insulin; porcine insulin; LysB28 ProB29 human insulin; GlyA21 ArgB31 ArgB32 human insulin; and LysB3 GluB29 human insulin or AspB28 desB30 human insulin.
  • acylated insulin is selected from the group consisting of N ⁇ B29 -(3- ⁇ 2-[2-(2-[ ⁇ -carboxy-pentadecanoyl- ⁇ -glutamyl-(2-amino-ethoxy)]-ethoxy)-ethoxy ⁇ -ethoxy]-propinoyl) desB30 human insulin, N ⁇ B29 -(3- ⁇ 2-[2-(2-[ ⁇ -carboxy-heptadecanoyl- ⁇ -glutamyl-(2-amino-ethoxy)]-ethoxy)-ethoxy ⁇ -ethoxy]-propinoyl) desB30 human insulin, N ⁇ B29 - ⁇ 3-[2-(2- ⁇ 2-[2-( ⁇ -carboxy-pentadecanoylamino)ethoxy]-ethoxy ⁇ -ethoxy)-ethoxy]-propionyl- ⁇ -gluta
  • composition according to any of the paragraphs 1-45 for the manufacture of a medicament for the treatment of diabetes.
  • 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 paragraphs 1-45 together with a pharmaceutically acceptable carrier.
  • 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 paragraphs 1-45 together with a pharmaceutically acceptable carrier.
  • a soluble pharmaceutical composition comprising an acylated insulin and further comprising more than 4 zinc atoms per 6 molecules of acylated insulin.
  • a soluble pharmaceutical composition according to paragraph 91 comprising an acylated insulin and further comprising more than 5 zinc atoms per 6 molecules of acylated insulin.
  • composition according to paragraphs 91-92 comprising up to about 14 zinc atoms per 6 molecules of acylated insulin.
  • composition according to paragraphs 91-93 comprising between about 5 and about 14 zinc atoms per 6 molecules of acylated insulin.
  • composition according to any of paragraphs 91-94 comprising between about 5 and about 13 zinc atoms per 6 molecules of acylated insulin.
  • composition according to any of paragraphs 91-95 comprising between about 5 and about 12 zinc atoms per 6 molecules of acylated insulin.
  • composition according to any of paragraphs 91-96 comprising between about 5.3 and about 12 zinc atoms per 6 molecules of acylated insulin.
  • composition according to any of paragraphs 91-97 comprising between about 5.5 and about 11.4 zinc atoms per 6 molecules of acylated insulin.
  • composition according to any of paragraphs 91-98 comprising between about 5.5 and about 10 zinc atoms per 6 molecules of acylated insulin.
  • composition according to any of paragraphs 91-99 wherein the acylated insulin is LysB29(N ⁇ -tetradecanoyl)desB30 human insulin or Lys B29 N ⁇ -lithocholoyl- ⁇ -Glu desB30 human insulin
  • W is:
  • X is:
  • Y is:
  • composition according to any of paragraphs 91-101 provided that the acylated insulin is not Lys B29 N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin.
  • composition according to any of the preceeding paragraphs having a pH between about 6.5 and 8.5
  • composition according to any of the preceding paragraphs further comprising a rapid acting insulin
  • composition according to paragraph 103, wherein the rapid acting insulin is AspB28 human insulin and/or LysB28ProB29 human insulin.
  • 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.
  • more than about 4.3 zinc atoms per 6 molecules of acylated insulin are added to the composition or more than about 4.5 zinc atoms per 6 molecules of acylated insulin are added to the composition or than about 5 zinc atoms per 6 molecules of acylated insulin are added to the composition
  • zinc atoms per 6 molecules of acylated insulin are added to the composition before the addition of a preservative or more preferred about 5 and about 11.4 zinc atoms per 6 molecules of acylated insulin are added to the composition before the addition of a preservative or even more preferred between about 5.5 and about 10 zinc atoms per 6 molecules of acylated insulin are added to the composition before the addition of a preservative
  • more than about 2 zinc atoms per 6 molecules of acylated insulin are added to the composition after the addition of a preservative or more than about 3 zinc atoms per 6 molecules of acylated insulin are added to the composition after the addition of a preservative or more than about 4 zinc atoms per 6 molecules of acylated insulin are added to the composition after the addition of a preservative
  • zinc atoms per 6 molecules of acylated insulin are added to the composition after the addition of a preservative or more preferred about 5 and about 11.4 zinc atoms per 6 molecules of acylated insulin are added to the composition after the addition of a preservative or even more preferred between about 5.5 and about 10 zinc atoms per 6 molecules of acylated insulin are added to the composition after the addition of a preservative
  • the method comprises adding at least 1 zinc atom per 6 molecules of acylated insulin before addition of a preservative and adding at least 1 zinc atom per 6 molecules of acylated insulin after addition of a preservative or adding at least 1 zinc atom per 6 molecules of acylated insulin before addition of a preservative and adding at least 2-3 zinc atoms per 6 molecules of acylated insulin after addition of a preservative or adding at least 1 zinc atom per 6 molecules of acylated insulin before addition of a preservative and up to about 11 zinc atom per 6 molecules of acylated insulin and after addition of a preservative
  • the method comprises adding at least 2 zinc atoms per 6 molecules of acylated insulin before addition of a preservative and adding at least 1 zinc atom per 6 molecules of acylated insulin after addition of a preservative or adding at least 2 zinc atoms per 6 molecules of acylated insulin before addition of a preservative and adding at least 2-3 zinc atoms per 6 molecules of acylated insulin after addition of a preservative or adding at least 2 zinc atoms per 6 molecules of acylated insulin before addition of a preservative and up to about 10 zinc atoms per 6 molecules of acylated insulin after addition of a preservative
  • the method comprises adding at least 3 zinc atoms per 6 molecules of acylated insulin before addition of a preservative and adding at least 1 zinc atom per 6 molecules of acylated insulin after addition of a preservative or adding at least 3 zinc atoms per 6 molecules of acylated insulin before addition of a preservative and adding at least 2-3 zinc atoms per 6 molecules of acylated insulin after addition of a preservative or adding at least 3 zinc atoms per 6 molecules of acylated insulin before addition of a preservative and up to about 9 zinc atoms per 6 molecules of acylated insulin after addition of a preservative
  • acylated insulin does not have a side chain attached to the ⁇ -amino group of a Lys residue present in the B chain of the parent insulin, the side chain being of the general formula:
  • W is:
  • X is:
  • Y is:
  • acylated insulin is not Lys B29 N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin.
  • Zn/Acyl-ins zinc atoms per 6 molecules of acylated insulin Acyl-ins acylated insulin
  • FIG. 1 Insulin aspart 600 ⁇ M with 3 zinc atoms per 6 molecules of insulin aspart
  • FIG. 2 Insulin aspart 600 ⁇ M with 6 zinc atoms per 6 molecules of insulin aspart
  • FIG. 3 Lys B29 N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin 600 ⁇ M with 3 zinc atoms per 6 molecules of acylated insulin
  • FIG. 5 Lys B29 N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin 600 ⁇ M with 6 zinc atoms per 6 molecules of acylated insulin
  • the present invention it is possible to design the insulin formulation containing the insulin with desired degree of association.
  • the invention provides soluble compositions that are mixtures of a rapid-acting insulin analogue and a prolonged-acting acylated insulin in which the rate of disappearance of the rapid acting insulin and the acylated insulin from the site of injection are the same as when injected in separate compositions.
  • the present invention is further based on the surprisingly recognition that, when preparing an insulin formulation, zinc can be added to the formulation after addition of preservatives.
  • the zinc is typically provided by adding zinc acetate, zinc chloride or zinc citrate to the insulin formulation.
  • an acylated insulin composition of this invention may be delivered by any of a variety of inhalation devices known in the art for administration of a therapeutic agent by inhalation. These devices include metered dose inhalers, nebulizers, dry powder generators, sprayers, and the like. Preferably, an acylated insulin composition of this invention is delivered by a dry powder inhaler or a sprayer.
  • an inhalation device for administering an acylated insulin composition of this invention is advantageously reliable, reproducible, and accurate.
  • the inhalation device should deliver small particles, for example, less than about 10 ⁇ m, for example about 1-5 ⁇ m, for good respirability.
  • the formulation of an acylated insulin composition of this invention depends on the type of inhalation device employed.
  • the frequency of administration and length of time for which the system is activated will depend mainly on the concentration of insulin conjugate in the aerosol. For example, shorter periods of administration can be used at higher concentrations of insulin conjugate in the nebulizer solution.
  • Devices such as metered dose inhalers can produce higher aerosol concentrations, and can be operated for shorter periods to deliver the desired amount of insulin conjugate.
  • Devices such as powder inhalers deliver active agent until a given charge of agent is expelled from the device. In this type of inhaler, the amount of insulin derivative of this invention in a given quantity of the powder determines the dose delivered in a single administration.
  • the particle size of insulin derivative of this invention in the formulation delivered by the inhalation device is critical with respect to the ability of insulin to make it into the lungs, and preferably into the lower airways or alveoli.
  • an acylated insulin composition of this invention is formulated so that at least about 10% of the insulin conjugate delivered is deposited in the lung, preferably about 10 to about 20%, or more. It is known that the maximum efficiency of pulmonary deposition for mouth breathing humans is obtained with particle sizes of about 2 ⁇ m to about 3 ⁇ m. When particle sizes are above about 5 ⁇ m pulmonary deposition decreases substantially. Particle sizes below about 1 ⁇ m cause pulmonary deposition to decrease, and it becomes difficult to deliver particles with sufficient mass to be therapeutically effective.
  • particles of an acylated insulin composition delivered by inhalation have a particle size preferably less than about 10 ⁇ m, more preferably in the range of about 1 ⁇ m to about 5 ⁇ m.
  • the formulation of the insulin derivative is selected to yield the desired particle size in the chosen inhalation device.
  • an acylated insulin composition of this invention is prepared in a particulate form with a particle size of less than about 10 ⁇ m, preferably about 1 to about 5 ⁇ m.
  • the preferred particle size is effective for delivery to the alveoli of the patient's lung.
  • the dry powder is largely composed of particles produced so that a majority of the particles have a size in the desired range.
  • at least about 50% of the dry powder is made of particles having a diameter less than about 10 ⁇ m.
  • Such formulations can be achieved by spray drying, milling, or critical point condensation of a solution containing insulin conjugate and other desired ingredients. Other methods also suitable for generating particles useful in the current invention are known in the art.
  • the particles are usually separated from a dry powder formulation in a container and then transported into the lung of a patient via a carrier air stream.
  • a carrier air stream typically, in current dry powder inhalers, the force for breaking up the solid is provided solely by the patient's inhalation.
  • air flow generated by the patient's inhalation activates an impeller motor which deagglomerates the particles.
  • Formulations of acylated insulin of this invention for administration from a dry powder inhaler typically include a finely divided dry powder containing the acylated insulin, but the powder can also include a bulking agent, carrier, excipient, another additive, or the like.
  • Additives can be included in a dry powder formulation of insulin conjugate, for example, to dilute the powder as required for delivery from the particular powder inhaler, to facilitate processing of the formulation, to provide advantageous powder properties to the formulation, to facilitate dispersion of the powder from the inhalation device, to stabilize the formulation (for example, antioxidants or buffers), to provide taste to the formulation, or the like.
  • the additive does not adversely affect the patient's airways.
  • the an acylated insulin can be mixed with an additive at a molecular level or the solid formulation can include particles of the insulin conjugate mixed with or coated on particles of the additive.
  • Typical additives include mono-, di-, and polysaccharides; sugar alcohols and other polyols, such as, for example, lactose, glucose, raffinose, melezitose, lactitol, maltitol, trehalose, sucrose, mannitol, starch, or combinations thereof; surfactants, such as sorbitols, diphosphatidyl choline, or lecithin; or the like.
  • an additive such as a bulking agent
  • an additive is present in an amount effective for a purpose described above, often at about 50% to about 90% by weight of the formulation.
  • Additional agents known in the art for formulation of a protein such as insulin analogue protein can also be included in the formulation.
  • a spray including the acylated insulin composition of this invention can be produced by forcing a suspension or solution of insulin conjugate through a nozzle under pressure.
  • the nozzle size and configuration, the applied pressure, and the liquid feed rate can be chosen to achieve the desired output and particle size.
  • An electrospray can be produced, for example, by an electric field in connection with a capillary or nozzle feed.
  • particles of insulin conjugate delivered by a sprayer have a particle size less than about 10 ⁇ m, preferably in the range of about 1 ⁇ m to about 5 ⁇ m.
  • Formulations of acylated insulin of this invention suitable for use with a sprayer will typically include the insulin derivative in an aqueous solution at a concentration of about 1 mg to about 20 mg of insulin conjugate per ml of solution.
  • the formulation can include agents such as an excipient, a buffer, an isotonicity agent, a preservative, a surfactant, and, preferably, zinc.
  • the formulation can also include an excipient or agent for stabilization of the insulin derivative, such as a buffer, a reducing agent, a bulk protein, or a carbohydrate.
  • Bulk proteins useful in formulating insulin conjugates include albumin, protamine, or the like.
  • Typical carbohydrates useful in formulating insulin conjugates include sucrose, mannitol, lactose, trehalose, glucose, or the like.
  • the insulin derivative formulation can also include a surfactant, which can reduce or prevent surface-induced aggregation of the insulin conjugate caused by atomization of the solution in forming an aerosol.
  • a surfactant which can reduce or prevent surface-induced aggregation of the insulin conjugate caused by atomization of the solution in forming an aerosol.
  • Various conventional surfactants can be employed, such as polyoxyethylene fatty acid esters and alcohols, and polyoxyethylene sorbitol fatty acid esters. Amounts will generally range between about 0.001 and about 4% by weight of the formulation.
  • compositions according to the present invention may be administered parenterally to patients in need of such a treatment.
  • Parenteral administration may be performed by subcutaneous, intramuscular or intravenous injection by means of a syringe, optionally a pen-like syringe or another convenient dosing equipment.
  • parenteral administration can be performed by means of an infusion pump.
  • One aspect of the invention is related to a pharmaceutical composition according to the invention together with a pharmaceutically acceptable carrier and/or a pharmaceutically acceptable additive, which can be provided for pulmonary treatment of type 1 diabetes, type 2 diabetes and other states that cause hyperglycaemia in patients in need of such a treatment.
  • a method for the manufacture of a pharmaceutical composition for the use in the treatment of type 1 diabetes, type 2 diabetes and other states that cause hyperglycaemia the composition being used pulmonary and comprising a therapeutically effective amount of a pharmaceutical composition according to the invention together with a pharmaceutically acceptable carrier and/or pharmaceutical acceptable additives.
  • Injectable compositions of the acylated insulin derivatives can be prepared using the conventional techniques of the pharmaceutical industry which involve dissolving and mixing the ingredients as appropriate to give the desired end product.
  • the insulin derivative is dissolved in an amount of water which is somewhat less than the final volume of the composition to be prepared.
  • An isotonic agent, a preservative or a mixture of preservatives, zinc as acetate, citrate or chloride or a mixture thereof, and a buffer can be added as required, furthermore a surfactant might be added and the pH value of the solution is adjusted—if necessary—using an acid, e.g. hydrochloric acid, or a base, e.g. aqueous sodium hydroxide as needed.
  • the volume of the solution is adjusted with water to give the desired concentration of the ingredients.
  • the buffer may be selected from the group consisting of sodium acetate, sodium carbonate, citrate, glycine, lysine, arginine, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium phosphate, ADA (N-[2-acetamido]-2-iminodiacetic acid), ACES (N-[2-acetamido]-2-aminoethanesulfonic acid), BES (N,N-bis[2-hydroxyethyl]-2-aminoethanesulfonic acid), bicine (N,N-bis-[2-hydroxyethyl]glycine), BIS-TRIS (bis[2-hydroxyethyl]iminotris[hydroxymethyl]-methane), DIPSO (3[N,N-bis(2-hydroxethyl]amin]-2-hydroxypropanesulfonic acid), ethylenediamine dihydrochloride, glycylglycine, HEPES (N-[2-hydroxyeth
  • the formulation further comprises a pharmaceutically acceptable preservative which may be selected from the group consisting of phenol, o-cresol, m-cresol, p-cresol, methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, 2-phenoxyethanol, butyl p-hydroxybenzoate, 2-phenylethanol, benzyl alcohol, chlorobutanol, and bronopol, benzoic acid, imidurea, chlorohexidine, sodium dehydroacetate, chlorocresol, ethyl p-hydroxybenzoate, benzethonium chloride, chlorphenesine (3-(4-chlorophenoxy)propane-1,2-diol) or mixtures thereof.
  • a pharmaceutically acceptable preservative which may be selected from the group consisting of phenol, o-cresol, m-cresol, p-cresol, methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, 2-phenoxyethanol, but
  • the preservative is present in a concentration from 0.1 mg/ml to 20 mg/ml. In a further aspect of the invention the preservative is present in a concentration from 0.1 mg/ml to 5 mg/ml. In a further aspect of the invention the preservative is present in a concentration from 5 mg/ml to 10 mg/ml. In a further aspect of the invention the preservative is present in a concentration from 10 mg/ml to 20 mg/ml. Each one of these specific preservatives constitutes an alternative aspect of the invention.
  • the use of a preservative in pharmaceutical compositions is well-known to the skilled person. For convenience reference is made to Remington: The Science and Practice of Pharmacy, 19 th edition, 1995.
  • the formulation further comprises an isotonic agent which may be selected from the group consisting of a salt (e.g. sodium chloride), a sugar or sugar alcohol, an amino acid (e.g. 1-glycine, 1-histidine, arginine, lysine, isoleucine, aspartic acid, tryptophan, threonine), 1,2-propanediol (propyleneglycol), 1,3-propanediol, 1,3-butanediol) polyethyleneglycol (e.g. PEG400), or mixtures thereof.
  • a salt e.g. sodium chloride
  • a sugar or sugar alcohol e.g. 1-glycine, 1-histidine, arginine, lysine, isoleucine, aspartic acid, tryptophan, threonine
  • 1,2-propanediol propyleneglycol
  • 1,3-propanediol 1,3-butanediol
  • Any sugar such as mono-, di-, or polysaccharides, or water-soluble glucans, including for example fructose, glucose, mannose, sorbose, xylose, maltose, lactose, sucrose, trehalose, dextran, pullulan, dextrin, cyclodextrin, soluble starch, hydroxyethyl starch and carboxymethylcellulose-Na may be used.
  • the sugar additive is sucrose.
  • Sugar alcohol is defined as a C4-C8 hydrocarbon having at least one —OH group and includes, for example, mannitol, sorbitol, inositol, galactitol, dulcitol, xylitol, and arabitol.
  • the sugar alcohol additive is mannitol.
  • the sugars or sugar alcohols mentioned above may be used individually or in combination. There is no fixed limit to the amount used, as long as the sugar or sugar alcohol is soluble in the liquid composition and does not adversely effect the stabilizing effects achieved using the methods of the invention.
  • the sugar or sugar alcohol concentration is between about 1 mg/ml and about 150 mg/ml.
  • the isotonic agent is present in a concentration from 1 mg/ml to 50 mg/ml. In a further aspect of the invention the isotonic agent is present in a concentration from 1 mg/ml to 7 mg/ml. In a further aspect of the invention the isotonic agent is present in a concentration from 8 mg/ml to 24 mg/ml. In a further aspect of the invention the isotonic agent is present in a concentration from 25 mg/ml to 50 mg/ml. Each one of these specific isotonic agents constitutes an alternative aspect of the invention.
  • the use of an isotonic agent in pharmaceutical compositions is well-known to the skilled person. For convenience reference is made to Remington: The Science and Practice of Pharmacy, 19 th edition, 1995.
  • the formulation comprises a surfactant to prevent fibrillation especially when mixing the insulin derivative with a rapid acting insulin as insulin aspart.
  • a surfactant to prevent fibrillation especially when mixing the insulin derivative with a rapid acting insulin as insulin aspart.
  • Various conventional surfactants can be employed, such as polyoxyethylene fatty acid esters and alcohols, and polyoxyethylene sorbitol fatty acid esters. Amounts will generally range between about 0.001 and about 0.1% by weight of the formulation.
  • Typical isotonic agents are sodium chloride, mannitol, dimethyl sulfone, 1,2 propandiol, and glycerol and typical preservatives are phenol, m-cresol, methyl p-hydroxybenzoate and benzyl alcohol.
  • buffers examples include sodium acetate, glycylglycine, HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), tris-hydroxymethyl-aminomethan, ethylenediamine dihydrochloride, and sodium phosphate.
  • the pharmaceutical compositions according to the present invention can be used in the treatment of states which are sensitive to insulin. Thus, they can be used in the treatment of type 1 diabetes, type 2 diabetes and hyperglycaemia for example as sometimes seen in seriously injured persons and persons who have undergone major surgery.
  • the optimal dose level for any patient will depend on a variety of factors including the efficacy of the specific acylated insulin or mixture of the acylated insulin with a rapid acting insulin employed, the age, body weight, physical activity, and diet of the patient, on a possible combination with other drugs, and on the severity of the state to be treated. It is recommended that the daily dosage of the insulin derivative of this invention be determined for each individual patient by those skilled in the art in a similar way as for known insulin compositions.
  • the starting product for preparing the acylated insulin or insulin analogue contained in the composition according to the invention can be produced by either well-known peptide synthesis or by well known recombinant production in suitable transformed microorganisms.
  • the insulin starting product can be produced by a method which comprises culturing a host cell containing a DNA sequence encoding the polypeptide and capable of expressing the polypeptide in a suitable nutrient medium under conditions permitting the expression of the peptide, after which the resulting peptide is recovered from the culture.
  • the medium used to culture the cells may be any conventional medium suitable for growing the host cells, such as minimal or complex media containing appropriate supplements. Suitable media are available from commercial suppliers or may be prepared according to published recipes (e.g. in catalogues of the American Type Culture Collection).
  • the peptide produced by the cells may then be recovered from the culture medium by conventional procedures including separating the host cells from the medium by centrifugation or filtration, precipitating the proteinaceous components of the supernatant or filtrate by means of a salt, e.g. ammonium sulphate, purification by a variety of chromatographic procedures, e.g. ion exchange chromatography, gel filtration chromatography, affinity chromatography, or the like, dependent on the type of peptide in question.
  • a salt e.g. ammonium sulphate
  • the DNA sequence encoding the insulin polypeptide may suitably be of genomic or cDNA origin, for instance obtained by preparing a genomic or cDNA library and screening for DNA sequences coding for all or part of the polypeptide by hybridisation using synthetic oligonucleotide probes in accordance with standard techniques (see, for example, Sambrook, J, Fritsch, E F and Maniatis, T, Molecular Cloning: A Laboratory Manual , Cold Spring Harbor Laboratory Press, New York, 1989).
  • the DNA sequence encoding the polypeptide may also be pre-pared synthetically by established standard methods, e.g.
  • the DNA sequence may be inserted into any vector which may conveniently be subjected to recombinant DNA procedures, and the choice of vector will often depend on the host cell into which it is to be introduced.
  • the vector may be an autonomously replicating vector, i.e. a vector which exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g. a plasmid.
  • the vector may be one which, when introduced into a host cell, is integrated into the host cell genome and replicated together with the chromosome(s) into which it has been integrated.
  • the vector is preferably an expression vector in which the DNA sequence encoding the peptide is operably linked to additional segments required for transcription of the DNA, such as a promoter.
  • the promoter may be any DNA sequence which shows transcriptional activity in the host cell of choice and may be derived from genes encoding proteins either homologous or heterologous to the host cell. Examples of suitable promoters for directing the transcription of the DNA encoding the peptide of the invention in a variety of host cells are well known in the art, cf. for instance Sambrook et al., Molecular Cloning—a laboratory manual, second edition 1989.
  • the DNA sequence encoding the peptide may also, if necessary, be operably connected to a suitable terminator, polyadenylation signals, transcriptional enhancer sequences, and translational enhancer sequences.
  • the recombinant vector of the invention may further comprise a DNA sequence enabling the vector to replicate in the host cell in question.
  • the vector may also comprise a selectable marker, e.g. a gene the product of which complements a defect in the host cell or one which confers resistance to a drug, e.g. ampicillin, kanamycin, tetracyclin, chloramphenicol, neomycin, hygromycin or methotrexate.
  • a selectable marker e.g. a gene the product of which complements a defect in the host cell or one which confers resistance to a drug, e.g. ampicillin, kanamycin, tetracyclin, chloramphenicol, neomycin, hygromycin or methotrexate.
  • a secretory signal sequence (also known as a leader sequence, prepro sequence or pre sequence) may be provided in the recombinant vector.
  • the secretory signal sequence is joined to the DNA sequence encoding the peptide in the correct reading frame.
  • Secretory signal sequences are commonly positioned 5′ to the DNA sequence encoding the peptide.
  • the secretory signal sequence may be that normally associated with the peptide or may be from a gene encoding another secreted protein.
  • the host cell into which the DNA sequence or the recombinant vector is introduced may be any cell which is capable of producing the present peptide and includes bacteria, yeast, fungi and higher eukaryotic cells.
  • suitable host cells well known and used in the art are, without limitation, E. coli, Saccharomyces cerevisiae , or mammalian BHK or CHO cell lines.
  • an effective amount means a dosage which is sufficient in order for the treatment of the patient to be effective compared with no treatment.
  • pharmaceutical composition means a product comprising an active compound or a salt thereof together with pharmaceutical excipients such as buffer, preservative and tonicity modifier, said pharmaceutical composition being useful for treating, preventing or reducing the severity of a disease or disorder by administration of said pharmaceutical composition to a person.
  • a pharmaceutical composition is also known in the art as a pharmaceutical formulation. It is to be understood that pH of a pharmaceutical composition which is to be reconstituted is the pH value which is measured on the reconstituted composition produced by reconstitution in the prescribed reconstitution liquid at room temperature.
  • pharmaceutically acceptable means suited for normal pharmaceutical applications, i.e. giving rise to no adverse events in patients etc.
  • buffer refers to a chemical compound in a pharmaceutical composition that reduces the tendency of pH of the composition to change over time as would otherwise occur due to chemical reactions. Buffers include chemicals such as sodium phosphate, TRIS, glycyl glycine and sodium citrate.
  • preservative refers to a chemical compound which is added to a pharmaceutical composition to prevent or delay microbial activity (growth and metabolism).
  • examples of pharmaceutically acceptable preservatives are phenol, m-cresol and a mixture of phenol and m-cresol.
  • isotonicity agent refers to a chemical compound in a pharmaceutical composition that serves to modify the osmotic pressure of the pharmaceutical composition so that the osmotic pressure becomes closer to that of human plasma.
  • Isotonicity agents include NaCl, glycerol, mannitol etc.
  • stabilizer refers to chemicals added to peptide containing pharmaceutical compositions in order to stabilize the peptide, i.e. to increase the shelf life and/or in-use time of such compositions.
  • stabilizers used in pharmaceutical formulations are L-glycine, L-histidine, arginine, polyethylene glycol, and carboxymethylcellulose. Further phenols, zinc ions and sodium chloride can act as stabilizers.
  • surfactant refers to a chemical compound in a pharmaceutical composition that serves to modify the interface to air and hydrophobic surfaces in a way that displaces or partly displaces insulin, insulin analogues and insulin derivatives from the interfaces.
  • Various conventional surfactants can be employed, such as polyoxyethylene fatty acid esters and alcohols, and polyoxyethylene sorbitol fatty acid esters. An example is polysorbate 20.
  • treatment of a disease means the management and care of a patient having developed the disease, condition or disorder.
  • the purpose of treatment is to combat the disease, condition or disorder.
  • Treatment includes the administration of the active compounds to eliminate or control the disease, condition or disorder as well as to alleviate the symptoms or complications associated with the disease, condition or disorder.
  • prevention of a disease is defined as the management and care of an individual at risk of developing the disease prior to the clinical onset of the disease.
  • the purpose of prevention is to combat the development of the disease, condition or disorder, and includes the administration of the active compounds to prevent or delay the onset of the symptoms or complications and to prevent or delay the development of related diseases, conditions or disorders.
  • human insulin as used herein means the human hormone whose structure and properties are well known. Human insulin has two polypeptide chains that are connected by disulphide bridges between cysteine residues, namely the A-chain and the B-chain.
  • the A-chain is a 21 amino acid peptide and the B-chain is a 30 amino acid peptide, the two chains being connected by three disulphide bridges: one between the cysteines in position 6 and 11 of the A-chain, the second between the cysteine in position 7 of the A-chain and the cysteine in position 7 of the B-chain, and the third between the cysteine in position 20 of the A-chain and the cysteine in position 19 of the B-chain.
  • basal insulin as used herein means an formulation of insulin peptide which has a time-action of more than 15 hours in standard models of diabetes and is suited to cover the need for insulin during the night and in-between meals.
  • the basal insulin has a time-action of at least 20 hours.
  • the basal insulin has a time-action of at least 10 hours.
  • the basal insulin has a time-action in the range from 15 to 48 hours.
  • the basal insulin has a time-action similar to that observed for commercial pharmaceutical compositions of NPH insulin or N ⁇ B29 -tetradecanoyl desB30 human insulin.
  • bolus insulin means an insulin peptide which is rapid-acting and suited to cover the need for insulin during and after the meal.
  • biphasic insulin as used herein means a pharmaceutical composition comprising a mixture of “bolus insulin” and “basal insulin”.
  • desB30 or “B(1-29)” is meant an insulin B chain or an analogue thereof lacking the B30 amino acid residue and “A(1-21)” means the natural insulin A chain or an analogue thereof.
  • the C-peptide and its amino acid sequence are indicated in the three letter amino acid code.
  • DesB30, desB29 human insulin is a human insulin lacking B29 and B30.
  • B1 the amino acid residue in position 1 in the B chain of insulin (counted from the N-terminal end) and the amino acid residue in position 1 in the A chain of insulin (counted from the N-terminal end), respectively.
  • the amino acid residue in a specific position may also be denoted as e.g. Phe B1 which means that the amino acid residue in position B1 is a phenylalanine residue.
  • insulin analogue as used herein is meant a polypeptide which has a molecular structure which formally can be derived from the structure of a naturally occurring insulin, for example that of human insulin by deleting and/or exchanging at least one amino acid residue in the naturally occurring insulin and/or adding at least one amino acid residue.
  • the added and/or exchanged amino acid residues can either be codable amino acid residues or other naturally occurring residues or purely synthetic amino acid residues
  • the insulin analogues may be such wherein position 28 of the B chain may be modified from the natural Pro residue to one of Asp, Lys, or Ile.
  • Lys at position B29 is modified to Pro.
  • B30 may be Lys and then B29 can be any codable amino acid except Cys, Met, Arg and Lys.
  • Asn at position A21 may be modified to Ala, Gln, Glu, Gly, His, Ile, Leu, Met, Ser, Thr, Trp, Tyr or Val, in particular to Gly, Ala, Ser, or Thr and preferably to Gly.
  • Asn at position B3 may be modified to Lys or Asp.
  • Further examples of insulin analogues are desB30 human insulin; desB30 human insulin analogues; insulin analogues wherein PheB1 has been deleted; insulin analogues wherein the A-chain and/or the B-chain have an N-terminal extension and insulin analogues wherein the A-chain and/or the B-chain have a C-terminal extension.
  • one or two Arg may be added to position B1 or to position B30.
  • an insulin analogue comprises less than 6 modifications (substitutions, deletions, additions) relative to the native peptide. In another aspect an analogue comprises less than 5 modifications (substitutions, deletions, additions) relative to the native peptide. In another aspect an analogue comprises less than 4 modifications (substitutions, deletions, additions) relative to the native peptide. In another aspect an analogue comprises less than 3 modifications (substitutions, deletions, additions) relative to the native peptide. In another aspect an analogue comprises less than 2 modifications (substitutions, deletions, additions) relative to the native peptide. In another aspect an analogue comprises only a single modification (substitutions, deletions, additions) relative to the native peptide.
  • insulin derivative as used herein is meant a naturally occurring insulin or an insulin analogue which has been chemically modified, e.g. by introducing a side chain in one or more positions of the insulin backbone or by oxidizing or reducing groups of the amino acid residues in the insulin or by converting a free carboxylic group to an ester group or acylating a free amino group or a hydroxy group.
  • acylated insulin as used herein is meant a naturally occurring insulin, for example that of human insulin, an insulin molecule, an insulin derivative or an insulin analogue which has been chemically modified, by acylating a free amino group or a hydroxy group.
  • nu blunting means that when formulated in one formulation both the rapid acting insulin and the acylated insulin has profile of action which is identical or substantially identical with the profile of action, when administering the rapid acting insulin and the acylated insulin in separate formulations.
  • OAD oral antidiabetic drug or oral antidiabetic drugs.
  • OAD(s) can be sulfonylurea (SU), biguanides e.g. Melformin or thiozolidindiones (TZD).
  • hGlu is homoglutamic acid.
  • ⁇ -Asp is the L-form of —HNCH(CO—)CH 2 COOH.
  • ⁇ -Asp is the L-form of —HNCH(COOH)CH 2 CO—.
  • ⁇ -Glu is the L-form of —HNCH(CO—)CH 2 CH 2 COOH.
  • ⁇ -Glu is the L-form of —HNCH(COOH)CH 2 CH 2 CO—.
  • ⁇ -hGlu is the L-form of —HNCH(CO—)CH 2 CH 2 CH 2 COOH.
  • ⁇ -hGlu is the L-form of —HNCH(COOH)CH 2 CH 2 CH 2 CO—.
  • Sar is sarcosine (N-methylglycine).
  • an amino acid residue having a carboxylic acid group in the side chain designates amino acid residues like Asp, Glu and hGlu.
  • the amino acids can be in either the L- or D-configuration. If nothing is specified it is understood that the amino acid residue is in the L configuration.
  • an amino acid residue having a neutral side chain designates amino acid residues like Gly, Ala, Val, Leu, Ile, Phe, Pro, Ser, Thr, Cys, Met, Tyr, Asn and Gln.
  • an insulin derivative according to the invention is stated to be “soluble at physiological pH values” it means that the insulin derivative can be used for preparing injectable insulin compositions that are fully dissolved at physiological pH values.
  • Such favourable solubility may either be due to the inherent properties of the insulin derivative alone or a result of a favourable interaction between the insulin derivative and one or more ingredients contained in the vehicle.
  • high molecular weight insulin means that the molecular weight of a complex of human insulin, of an insulin analogue or of an insulin derivative is above human serum albumin, above a dodecameric complex of an insulin analogue or of an insulin derivative or more than about 72 kDalton.
  • medium molecular weight insulin means that the molecular weight of a complex of human insulin, of an insulin analogue or of an insulin derivative is from about an insulin hexamer to about an insulin dodecamer between 24 and 80 kDalton
  • low molecular weight insulin or “lmw” means that the molecular weight of a human insulin, an insulin analogue or an insulin derivative is below 24 kDalton
  • net charge means the overall charge of the molecule. At pH 7.4, human insulin has a negative net charge about ⁇ 3 or when forming a hexamer about ⁇ 2.5 per insulin monomer.
  • LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin 1200 ⁇ M, 3 Zn/6ins, 1.6% glycerol, 16 mM phenol and 16 mM m-cresol, 7 mM trishydroxymethylaminomethan, 10 mM NaCl, pH 7.5 and volume of 7 mL: 58 mg LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin was suspended and dissolved in 2 mL water and subsequent added 420 ⁇ L 10 mM Zn(AcO) 2 , 2800 ⁇ L 4% glycerol, 350 ⁇ L 0.32 M phenol, 700 ⁇ L m-cresol, 490 ⁇ L 0.1 M trishydroxymethylaminomethan, 140 ⁇ L 0.5 M NaCl and finally addition of 1 N NaOH adjusting to pH 7.5 and water to obtain 7 mL.
  • D LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin 1200 ⁇ M, 4 Zn/6ins, 1.6% glycerol, 16 mM phenol and 16 mM m-cresol, 7 mM trishydroxymethylaminomethan, 10 mM NaCl, pH 7.5 and volume of 20 mL:
  • the formulation was prepared analogous to formulation B.
  • E Insulin aspart 600 ⁇ M, 6 Zn/6ins, 1.6% glycerol, 16 mM phenol and 16 mM m-cresol, 7 mM trishydroxymethylaminomethan, 10 mM NaCl, pH 7.5: The formulation was prepared as C and finally adding 20 ⁇ L 10 mM Zn(AcO) 2 per mL and adjusting pH to 7.5.
  • G Insulin aspart 600 ⁇ M, 8 Zn/6ins, 1.6% glycerol, 16 mM phenol and 16 mM m-cresol, 7 mM trishydroxymethylaminomethan, 10 mM NaCl, pH 7.5:
  • the formulation was prepared as C and finally adding 40 ⁇ L 10 mM Zn(AcO) 2 per mL and adjusting pH to 7.5
  • H LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin 1200 ⁇ M, 8 Zn/6ins, 1.6% glycerol, 16 mM phenol and 16 mM m-cresol, 7 mM trishydroxymethylaminomethan, 10 mM NaCl, pH 7.5:
  • the formulation was prepared as D and finally adding 80 ⁇ L 10 mM Zn(AcO) 2 per mL and adjusting pH to 7.5.
  • Formulations of LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin 600 ⁇ M and Zn/6 acyl-ins varied at 3, 4, 6, and 8 were obtained by dilution of the above mentioned formulation B, D, F, and H with an equal amount of medium.
  • the formulations were tested by size exclusion chromatography for the ability to form high molecular weight insulin on a Superose 6HR column eluted with 140 mM NaCl, 10 mM trishydroxymethylaminomethan pH 7.4, and 0.01% NaN 3 at 37° C. and 0.25 mL/min.
  • Fractions were collected for every 4 minutes starting at the high molecular weight exclusion limit and ending with the last of the monomer peak, in total 14 fractions.
  • the fraction concentration of insulin analogue was quantified specifically by reverse phase chromatography and the concentration of zinc after addition of the chromophoric zinc chelator terpy at an pH 2.5 excursion (FIGS. 3 , 4 , 5 and 6 ).
  • the relative amount of LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin with a size larger than albumin (fraction [1-8]) was measured at 3, 4, 6, and 8 Zn/6 insulin to 67.4, 88.9, 98.0, and 97.9% respectively.
  • the zinc concentration was following LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin especially in the high molecular size fractions.
  • Formulation consisting of insulin aspart 600 ⁇ M (A and E) varied at 3 and 6 zinc/insulin eluted as monomer (fraction[12-13]) only followed by minor part of the added zinc, see FIGS. 1-2 .
  • Formulations consisting of insulin 180 ⁇ M aspart and 420 ⁇ M LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin and zinc concentration varied at 3, 6, and 8 Zn/6insulin were obtained by mixing the above stock solution A to H and the mentioned medium in an analogous manner.
  • the formulations were tested by size exclusion chromatography for the ability to form high molecular weight insulin on a Superose 6HR column, and fractions were similarly collected for quantification of both insulin analogues and zinc.
  • the analogues mixing with 3 Zn/6insulin corresponding to a formulation of 4.3 Zn/6 acyl-insulin showed predominantly LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin (72.5% of LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin) and zinc in the high molecular weight fractions [1-8], but also insulin aspart, whereas an equal amount of insulin aspart and LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin was measured as mean in the rest of the fractions [9-14]( FIG.
  • LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin was dissolved in 400 ⁇ L water and 3 ⁇ Ci 125 I-Tyr A14 -LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin tracer was added, followed by 800 ⁇ L 4% glycerol and treated for 10-15 min in a vacuum centrifuge to remove added ethanol included in the tracer solution.
  • LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin was dissolved in 400 ⁇ L water and 3 ⁇ Ci 125 I-Tyr A14 -LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin tracer was added, followed by 800 ⁇ L 4% glycerol and treated for 10-15 min in a vacumcentrifuge to remove added ethanol included in the tracer solution.
  • 60 ⁇ L 10 mM Zn(AcO) 2 was added and with 2 minutes interval followed by 100 ⁇ L 0.32 M phenol, 200 ⁇ L 0.16 M m-cresol, 60 ⁇ L 10 mM Zn(AcO) 2 , 80 ⁇ L 0.5 M NaCl, and 140 ⁇ L 0.1 M sodium phosphate and adjusted to pH 7.5 mM and a total volume of 2 mL.
  • the solution was filtered through a sterile 0.22 ⁇ m filter and used for a disappearance study after subcutaneous injection in pig.
  • T 50% ⁇ SEM (h) was determined to 11.9 ⁇ 1.0 h as described under assay in a cross over study of formulation examples 2A to 2D.
  • LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin was dissolved in 400 ⁇ L water and 3 ⁇ Ci 125 I-Tyr A14 -LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin tracer was added, followed by 800 ⁇ L 4% glycerol and treated for 10-15 min in a vacumcentrifuge to remove added ethanol included in the tracer solution.
  • LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin was dissolved in 400 ⁇ L water and 3 ⁇ Ci 125 I-Tyr A14 -LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin tracer was added, followed by 800 ⁇ L 4% glycerol and treated for 10-15 min in a vacuum centrifuge to remove added ethanol included in the tracer solution.
  • LysB29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 human insulin can be crystallized as a hexamer with a zinc content of 2-6 mol zinc per 6 mol insulin and an excess of a phenol-like molecule (preferred phenol).
  • a phenol-like molecule preferred phenol
  • precipitants ionic salts are used with NaCl being preferred.
  • crystallisation can be enhanced by adding small amounts of organic solvents (ethanol).
  • B29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 insulin (to 600 ⁇ M) was suspended in water and dissolved by addition of sodium hydroxide, and added in following order: glycerol 1.6%, phenol and 16 mM m-cresol 16 mM, zinc acetate 0 to 6 Zn/6ins, sodium chloride 10 mM, phosphate 7 mM, pH 7.5, and water to volume adjustment.
  • K AV ( t ⁇ t 0 )/( V/f+t d ⁇ t 0 ) with
  • K AV peak 1 about 0.56 (albumin size) integration cut is between albumin size and insulin hexamer size.
  • peak1 For SEC method without phenol in eluent peak1 contain associated forms larger than albumin and peak2 contain dihexameric, hexameric, dimeric and monomeric forms of insulin.
  • B29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 insulin 600 ⁇ M was formulated with zinc concentrations increasing stepwise by one from 0 to 6 Zn/6ins, and the SEC method showed self association to very high molecular weight and more than human albumin for more than 90%, when the zinc concentration was 5 Zn/6ins. Including phenol in the SEC eluent two sharp peaks were seen, at insulin dihexamer size and insulin monomer size, and at 4 Zn/6ins more than 95% was at dihexamer size.
  • B29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 insulin (ins.deriv.) (to 1200 ⁇ M) was suspended in water, dissolved and added glycerol 1.6%, and 16 mM phenol and 16 mM m-cresol. Before zinc addition pH was adjusted to 7.5 by sodium hydroxide, and zinc acetate added in smaller portions of max 1 Zn/6ins up to 3 Zn/6ins (ad 1), to 5.62 Zn/6ins (ad II), and to 6 Zn/6ins (III). Sodium chloride was then added to 10 mM followed by adjustment of pH to 7.5 by sodium hydroxide, and adjustment of volume by water.
  • Insulin aspart (to 600 ⁇ M) was suspended in water and added hydrochloric acid to about pH 2.5, zinc acetate to 3 Zn/6ins, glycerol to 1.6%, 16 mM phenol and 16 mM m-cresol to sodium chloride to 10 mM, pH to 7.5 and water to final volume (IV).
  • insulin aspart (IV) was mixed to B29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 insulin in molar relation of 1:8 and total zinc concentration of 3.38 Zn/6ins.deriv.(I)(lowZnmix) and 6 Zn/6ins.deriv.(II)(highZnmix).
  • each pig received sc in random order either a lowZnmix (I) or a highZnmix (II) or the two analogues (III and IV) administered separately in the same pig.
  • the doses were 0.9 nmol/kg of insulin aspart and 7.2 nmol/kg of B29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 insulin.
  • the pigs were kept euglycaemic at their individual fasting glucose levels by infusion of a 20% glucose solution. Dependent on changes in plasma glucose concentration adjustments of glucose infusion rate were made empirically. Blood samples were collected 0-24 h for specific ELISA plasma analysis of immunoreactive insulin, and the pharmacokinetic profiles are shown in FIG. 13 and FIG. 14 .
  • B29N ⁇ -hexadecandioyl- ⁇ -Glu des(B30) insulin (ins.deriv.) (to 600 ⁇ M) was suspended in water, dissolved and added glycerol 1.6%, phenol 16 mM, and m-cresol 16 mM. pH was adjusted to 7.5 by sodium hydroxide, and citrate was added in three formulations to 0.6, 1.8 and 6 mM respectively in connection with zinc acetate to 6 zinc/6 insulin deriv. Sodium chloride was then added to 10 mM and sodium phosphate (pH 7.5) to 5 mM followed by adjustment of pH to 7.5 or 7.8 by sodium hydroxide, and adjustment of volume by water.
  • the formulations were studied after storage 2 week at 37 C compared to storage at 5 C by the SEC method described in example 4 using an eluent with 2 mM phenol. Reference formulations without citrate at 3, 5 and 6 Zn/6ins.deriv. were included.
  • the long acting insulin derivative is predominantly associated at a size of albumin corresponding to a dihexameric form (and not higher associated forms), and the amount of insulin derivative in the monomeric form is decreased at zinc concentration increased from 3 to 5 and 6 Zn/6ins.deriv.
  • Adding 1, 3 or 10 citrate equivalents to the zinc concentration at 6 Zn/6ins.deriv. showed less content of the monomeric insulin derivative compared to a reference at 3 Zn/ins.deriv.
  • the self associating pattern was not changed after storage 2 weeks at 37° C.
  • Insulin Aspart 3Zn/6ins
  • insulin derivative formulations comprising citrate at three levels and 6 Zn/6ins were shown to be mixable with insulin aspart after 2 weeks storage at 37 C, whereas insulin aspart was partly included in the high molecular weight fraction at the normal level of zinc at 3 Zn/6ins.
  • B29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 insulin (ins.deriv.) (to 600 ⁇ M) is suspended in water, dissolved, (if needed by addition of sodium hydroxide), and added glycerol 1.6%, phenol 16 mM, and m-cresol 16 mM. pH is adjusted to 7.5 by sodium hydroxide, and zinc citrate is added to 0.6 mM Zinc ion. Sodium chloride is then added to 10 mM and sodium phosphate (pH 7.5) to 5 mM followed by adjustment of pH to 7.5 and adjustment of volume by water.
  • B29N ⁇ -hexadecandioyl- ⁇ -Glu desB30 insulin (ins.deriv.) (to 600 ⁇ M) is suspended in water, dissolved and added glycerol 1.6%, and 16 mM phenol and 16 mM m-cresol. pH is adjusted to 7.5 by sodium hydroxide, and zinc acetate to 6 zinc/6 insulin deriv. (optionally zinc as citrate).
  • Sodium chloride is then added to 10 mM, a surfactant eg. poloxamer 188 or polysorbate 20 to about 0.002% and sodium phosphate (pH 7.5) to 5 mM followed by adjustment of pH to 7.5 and adjustment of volume by water.
  • Table 3 shows SEC, measured as described in example 4 with 2 or 6 Zn(II) per 6 insulins.
  • Hexadecanedioic acid (20.0 g, 69.8 mmol), n-octane and Dowex® are suspended and heated to reflux.
  • Benzyl formate (22.0 g, 162 mmol) is added.
  • additional benzyl formate (22.0 g, 162 mmol) is added.
  • the heating is continued for 50 hours.
  • the reaction mixture is filtered at 80° C.
  • the filtrate is cooled to 20° C., and the precipitate is collected by filtration.
  • the crude product (20.2 g) is suspended in dichloromethane (220 ml) at 20° C. for 4 hours.
  • the suspension is filtered, and the filtrate is evacuated to dryness at 20-30° C.
  • the resulting solid (13.9 g) is recrystallised from 2-propanol (140 ml).
  • 1,16-hexadecanedioic acid mono benzyl ester (20.0 g, 53.1 mmol) is dissolved in acetone at 35-40° C. N-Hydroxysuccinimide (6.42 g/55.8 mmol) is added. To the resulting solution dicyclohexylcarbodiimide (DCC) (12.1 g/58.4 mmol) is added. The reaction mixture is stirred for 3-4 hours at 35° C. To the resulting suspension triethylamine (7.40 ml, 53.1 mmol) og L-glutamic acid ⁇ -benzyl ester (12.6 g/53.1 mmol) are added.
  • DCC dicyclohexylcarbodiimide
  • the reaction mixture is stirred for 8-16 hours at 35-40° C.
  • the reaction mixture is cooled to 20-25° C.
  • Methanesulfonic acid (3.45 ml/53.1 mmol) and DCC (12.1 g/53.1 mmol) are added.
  • the reaction mixture is stirred for 8-16 hours at 20-25° C.
  • the reaction mixture is filtered, and the filtrate evacuated to dryness.
  • the residue is partitioned between water (100 ml) and toluene (200 ml).
  • the toluene phase is dried by distilling of water.
  • Silica gel (20 g) is added to the residue.
  • the suspension is stirred for 30 minutes at 20-25° C., then filtered.
  • the volume of the filtrate is reduced to ca 100-120 ml by evaporation under reduced pressure.
  • N-heptane 150 ml is added over a period of 15-30 minutes.
  • the resulting suspension is stirred for 2 hours.
  • the product is isolated by filtration, and dried to constant weight under reduced pressure at 20-25° C. Yield 21 g (58%) of white material.
  • PC2414 448 mg is dissolved in 3.5 g NMP (N-methyl-2-pyrrolidon) stabilised with 10 ⁇ l 5% sulphuric acid.
  • the desB30 human insulin solution is stirred and the PC2414 solution is added over a 20 min period, while keeping the temperature low.
  • reaction mixture is diluted with 2.5 weight of a solution consisting of: Tris-hydroxymethylaminomethan (20 mmol/kg), Ammonium acetate (30 mmol/kg), ethanol 42.5% w/w, the rest purified water, pH 7.5
  • the pH is adjusted to 7.5 by slowly adding 1 M acetic acid, while stirring.
  • Lys B29 N ⁇ -hexadecandioyl- ⁇ -glutamyl
  • desB30 human insulin emerged from the column after about 6 min.
  • Sprague Dawley male rats weighing 238-383 g on the experimental day were used for the clamp experiment.
  • the rats had free access to feed under controlled ambient conditions and were fasted overnight (from 3 pm) prior to the clamp experiment.
  • the rats were acclimatized in the animal facilities for at least 1 week prior to the surgical procedure. Approximately 1 week prior to the clamp experiment Tygon catheters were inserted under halothane anaesthesia into the jugular vein (for infusion) and the carotid artery (for blood sampling) and exteriorised and fixed on the back of the neck. The rats were given Streptocilin vet. (Boehringer Ingelheim; 0.15 ml/rat, i.m.) post-surgically and placed in an animal care unit (25° C.) during the recovery period.
  • Anorphin (0.06 mg/rat, s.c.) was administered during anaesthesia and Rimadyl (1.5 mg/kg, s.c.) was administered after full recovery from the anaesthesia (2-3 h) and again once daily for 2 days.
  • Plasma glucose levels were measured at 10 min intervals throughout and infusion of 20% aqueous glucose was adjusted accordingly in order to maintain euglyceamia. Samples of re-suspended erythrocytes were pooled from each rat and returned in about 1 ⁇ 2 ml volumes via the carotid catheter.
  • T 50% is the time when 50% of an injected amount of the A14 Tyr[ 125 I] labelled derivative of an insulin to be tested has disappeared from the injection site as measured with an external ⁇ -counter.
  • Formulated preparations of insulin derivatives labelled in Tyr A14 with 125 I were injected sc. in pigs as previously described (Ribel, U., Jorgensen, K, Brange, J, and Henriksen, U. The pig as a model for subcutaneous insulin absorption in man. Serrano-Rios, M and Lefèbvre, P. J. 891-896. 1985. Amsterdam; N.Y.; Oxford, Elsevier Science Publishers. 1985 (Conference Proceeding)).
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US20090137454A1 (en) * 2006-02-27 2009-05-28 Novo Nordisk A/S Insulin Derivatives
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US20080171695A1 (en) 2005-02-02 2008-07-17 Novo Nordisk A/S Insulin Derivatives
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US20130331320A1 (en) 2010-12-14 2013-12-12 Novo Nordisk A/S Fast-acting insulin in combination with long-acting insulin
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Citations (35)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3528960A (en) * 1968-10-07 1970-09-15 Lilly Co Eli N-carboxyaroyl insulins
US3868358A (en) * 1971-04-30 1975-02-25 Lilly Co Eli Protamine-insulin product
US4476118A (en) * 1982-02-05 1984-10-09 Novo Industri A/S Stabilized insulin preparations
US5177058A (en) * 1988-08-13 1993-01-05 Hoechst Aktiengesellschaft Pharmaceutical formulation for the treatment of diabetes mellitus
US5382574A (en) * 1989-12-21 1995-01-17 Novo Nordisk A/S Insulin preparations containing nicotinic acid or nicotinamide
US5646242A (en) * 1994-11-17 1997-07-08 Eli Lilly And Company Selective acylation of epsilon-amino groups
US5750497A (en) * 1993-09-17 1998-05-12 Novo Nordisk A/S Acylated insulin
US5830999A (en) * 1995-01-26 1998-11-03 Regents Of The University Of California Stabilization of insulin through ligand binding interations
US5866538A (en) * 1996-06-20 1999-02-02 Novo Nordisk A/S Insulin preparations containing NaCl
US5898067A (en) * 1997-02-07 1999-04-27 Novo Nordisk A/S Crystallization of proteins
US5905140A (en) * 1996-07-11 1999-05-18 Novo Nordisk A/S, Novo Alle Selective acylation method
US6011007A (en) * 1993-09-17 2000-01-04 Novo Nordisk A/S Acylated insulin
US6174856B1 (en) * 1998-01-09 2001-01-16 Novo Nordisk A/S Stabilized insulin compositions
US6211144B1 (en) * 1998-10-16 2001-04-03 Novo Nordisk A/S Stable concentrated insulin preparations for pulmonary delivery
US6251856B1 (en) * 1995-03-17 2001-06-26 Novo Nordisk A/S Insulin derivatives
US6335316B1 (en) * 1997-10-31 2002-01-01 Eli Lilly And Company Method for administering acylated insulin
US20020045731A1 (en) * 1996-02-21 2002-04-18 Novo Nordisk Of North America, Inc. Peptide derivatives
US6451762B1 (en) * 1997-10-24 2002-09-17 Novo Nordisk A/S Aggregates of human insulin derivatives
US20020155994A1 (en) * 1997-10-24 2002-10-24 Svend Havelund Aggregates of human insulin derivatives
US20030004096A1 (en) * 2001-03-23 2003-01-02 Peter Boderke Zinc-free and low-zinc insulin preparations having improved stability
US6504005B1 (en) * 1996-08-07 2003-01-07 Yeda Research And Development Co. Ltd. Long-acting drugs and pharmaceutical compositions comprising them
US6531448B1 (en) * 1997-12-23 2003-03-11 Eli Lilly And Company Insoluble compositions for controlling blood glucose
US6652886B2 (en) * 2001-02-16 2003-11-25 Expression Genetics Biodegradable cationic copolymers of poly (alkylenimine) and poly (ethylene glycol) for the delivery of bioactive agents
US20040006000A1 (en) * 2002-05-07 2004-01-08 Liselotte Langkjaer Novel formulations
US20040138099A1 (en) * 2002-11-29 2004-07-15 Draeger Eberhard Kurt Insulin administration regimens for the treatment of subjects with diabetes
US6869930B1 (en) * 1993-09-17 2005-03-22 Novo Nordisk A/S Acylated insulin
US20060183668A1 (en) * 2003-08-05 2006-08-17 Novo Nordisk A/S Novel insulin derivatives
US20080076705A1 (en) * 2005-02-02 2008-03-27 Kodra Janos T Insulin Derivatives
US7402565B2 (en) * 2003-11-14 2008-07-22 Novo Nordisk A/S Processes for making acylated insulin
US20090137454A1 (en) * 2006-02-27 2009-05-28 Novo Nordisk A/S Insulin Derivatives
US7544656B2 (en) * 2003-03-04 2009-06-09 The Technology Development Company, Ltd. Long acting injectable insulin composition and methods of making and using thereof
US20090239785A1 (en) * 2006-05-09 2009-09-24 Novo Nordisk A/S Insulin Derivative
US20100167990A1 (en) * 2007-06-13 2010-07-01 Novo Nordisk A/S Pharmaceutical Formulations Comprising an Insulin Derivative
US20110152185A1 (en) * 2007-11-16 2011-06-23 Novo Nordisk A/S Pharmaceutical Compositions Comprising GLP-1 Peptides or Extendin-4 and a Basal Insulin Peptide
US8003605B2 (en) * 2002-03-13 2011-08-23 Novo Nordisk A/S Minimising body weight gain in insulin treatment

Family Cites Families (104)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE1212679B (de) 1957-08-03 1966-03-17 Novo Terapeutisk Labor As Verfahren zur Herstellung von Insulinloesungen
GB1042194A (en) 1962-04-30 1966-09-14 Olin Mathieson Insulin preparations
US3907676A (en) 1970-07-28 1975-09-23 Novo Terapeutisk Labor As Process for purifying insulin
GB1492997A (en) 1976-07-21 1977-11-23 Nat Res Dev Insulin derivatives
JPS5767548A (en) 1980-10-14 1982-04-24 Shionogi & Co Ltd Insulin analog and its preparation
US4652548A (en) 1981-08-27 1987-03-24 Eli Lilly And Company Pharmaceutical formulations comprising human insulin, human C-peptide, and human proinsulin
US4876322A (en) 1984-08-10 1989-10-24 Siemens Aktiengesselschaft Irradiation cross-linkable thermostable polymer system, for microelectronic applications
DE3562922D1 (en) 1984-08-17 1988-06-30 Bayer Ag Thermoplastic moulding masses with flame-retarding properties
DE3433367A1 (de) 1984-09-12 1986-03-20 Robert Bosch Gmbh, 7000 Stuttgart Einrichtung zur steuerung der energiezufuhr zu einer heissen stelle
US5008241A (en) 1985-03-12 1991-04-16 Novo Nordisk A/S Novel insulin peptides
DK113585D0 (da) 1985-03-12 1985-03-12 Novo Industri As Nye peptider
US4683202A (en) 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
JPS61226115A (ja) 1985-03-30 1986-10-08 Mitsubishi Metal Corp 線条体押し出し機
PH25772A (en) 1985-08-30 1991-10-18 Novo Industri As Insulin analogues, process for their preparation
US4764592A (en) 1987-04-23 1988-08-16 Eli Lilly And Company Crystalline human proinsulin and process for its production
US5605884A (en) 1987-10-29 1997-02-25 Rhone-Poulenc Rorer Pharmaceuticals Inc. Factor VIII formulations in high ionic strength media
US4877608A (en) 1987-11-09 1989-10-31 Rorer Pharmaceutical Corporation Pharmaceutical plasma protein formulations in low ionic strength media
JPH01254699A (ja) 1988-04-05 1989-10-11 Kodama Kk インスリン誘導体及びその用途
KR910700262A (ko) 1988-12-23 1991-03-14 안네 제케르 사람 인슐린 유사체
NZ232375A (en) 1989-02-09 1992-04-28 Lilly Co Eli Insulin analogues modified at b29
IT1240314B (it) 1989-09-28 1993-12-07 Immunobiology Research Institutes, Inc. Formulazioni acquose stabilizzate di piccoli peptidi.
DK45590D0 (zh) 1990-02-21 1990-02-21 Novo Nordisk As
DK175491D0 (da) 1991-10-18 1991-10-18 Novo Nordisk As Apparat
ATE227583T1 (de) 1991-12-20 2002-11-15 Novo Nordisk As Stabilisierte pharmazeutische formulierung, die wachstumshormon und histidin enthält
US5672581A (en) 1993-01-29 1997-09-30 Aradigm Corporation Method of administration of insulin
US5652216A (en) 1994-05-26 1997-07-29 Novo Nordisk A/S Pharmaceutical preparation
WO1996010417A1 (en) 1994-10-04 1996-04-11 Novo Nordisk A/S PREPARATIONS CONTAINING AspB28 HUMAN INSULIN AND NICOTINAMIDE
US5693609A (en) 1994-11-17 1997-12-02 Eli Lilly And Company Acylated insulin analogs
BR9607647A (pt) 1995-03-17 1999-04-06 Novo Nordisk As Derivado de insulina e composição farmacêutica e processo para o tratamento de diabetes em um paciente em necessidade deste tratamento
CA2226575C (en) 1995-07-27 2011-10-18 Genentech, Inc. Stabile isotonic lyophilized protein formulation
EP0938502B1 (en) 1996-07-11 2004-10-06 Novo Nordisk A/S Selective acylation method
US5763401A (en) 1996-07-12 1998-06-09 Bayer Corporation Stabilized albumin-free recombinant factor VIII preparation having a low sugar content
ES2218622T3 (es) 1996-07-26 2004-11-16 Aventis Pharma Deutschland Gmbh Derivados de insulina con actividad de union al zinc incrementada.
WO1998042368A1 (en) 1997-03-20 1998-10-01 Novo Nordisk A/S Therapeutic powder formulation for pulmonary administration, containing crystalline insulin
EP0969860B1 (en) * 1997-03-20 2002-07-17 Novo Nordisk A/S Method for preparation of a therapeutic powder through coprecipitation of insulin and absorption enhancer
US7097845B2 (en) 1997-04-23 2006-08-29 Jacob Sten Petersen Combinations of antigen and mucosal binding component for inducing specific immunological tolerance
EP1283051B1 (en) 1997-06-13 2006-06-14 Eli Lilly And Company Stable insulin formulations
ZA984697B (en) 1997-06-13 1999-12-01 Lilly Co Eli Stable insulin formulations.
PL195845B1 (pl) 1997-10-24 2007-10-31 Novo Nordisk As Rozpuszczalny w wodzie agregat pochodnych insuliny
EP1039920A4 (en) 1997-10-24 2003-05-28 Lilly Co Eli INSULIN ANALOGS ACYLATED BY FATTY ACID
CN1278737A (zh) 1997-11-12 2001-01-03 阿尔萨公司 减少多肽自结合的方法
DK1121144T3 (da) * 1998-10-16 2002-09-23 Novo Nordisk As Stabile koncentrerede insulinpræparater til pulmonar indgivelse
ES2180511T3 (es) * 1999-01-26 2003-02-16 Lilly Co Eli Formulaciones monodispersas de analogos de insulina acilados hexamericos.
PT2130554E (pt) 1999-02-22 2012-11-19 Univ Connecticut Formulações de factor viii isentas de albumina
AU4450700A (en) 1999-04-27 2000-11-10 Eli Lilly And Company Insulin crystals for pulmonary administration
GB9930882D0 (en) 1999-12-30 2000-02-23 Nps Allelix Corp GLP-2 formulations
DE10022092A1 (de) 2000-05-08 2001-11-15 Aventis Behring Gmbh Stabilisiertes Protein-Präparat und Verfahren zu seiner Herstellung
JP4624600B2 (ja) * 2001-06-08 2011-02-02 トヨタ自動車株式会社 焼結合金、その製造方法およびバルブシート
DE60224284T2 (de) 2001-06-28 2008-12-18 Novo Nordisk A/S Stabile formulierung von modifiziertem glp-1
JP2004537580A (ja) 2001-08-10 2004-12-16 エピックス メディカル， インコーポレイテッド 延長された循環半減期を有するポリペプチド結合体
US20030068361A1 (en) 2001-10-09 2003-04-10 Rimona Margalit Liposome-encapsulated insulin formulations
EP1545460A4 (en) 2001-12-20 2005-11-16 Lilly Co Eli INSULIN MOLECULE WITH TEMPORARY EFFECT
AU2003218635A1 (en) 2002-05-07 2003-11-11 Novo Nordisk A/S Soluble formulations comprising insulin aspart and insulin detemir
US20050232899A1 (en) 2002-05-31 2005-10-20 Aradigm Corporation Compositions methods and systems for pulmonary delivery of recombinant human interferon alpha-2b
US20040002451A1 (en) 2002-06-20 2004-01-01 Bruce Kerwin Compositions of pegylated soluble tumor necrosis factor receptors and methods of preparing
KR100615389B1 (ko) 2002-08-23 2006-08-25 (주)헬릭서 다래 추출물을 함유하는 알러지성 질환 및 비알러지성염증 질환의 예방 및 개선용 건강 기능 식품
JP2006504772A (ja) 2002-10-29 2006-02-09 アルザ・コーポレーション 安定化された固体ポリペプチド粒子
MXPA05008363A (es) 2003-02-07 2005-12-06 Ajinomoto Kk Agentes terapeuticos para diabetes.
EP1597367B1 (en) 2003-02-19 2010-07-07 Novartis AG Glycoprotein antigen sima135 expressed in metastatic human tumor cells
ES2382157T3 (es) 2003-06-25 2012-06-05 Novo Nordisk Health Care Ag Composición líquida de polipépttidos del factor VII
US20050054818A1 (en) 2003-07-02 2005-03-10 Brader Mark Laurence Crystalline compositions for controlling blood glucose
WO2005005477A2 (en) 2003-07-11 2005-01-20 Novo Nordisk A/S Stabilised insulin compositions
PT2107069E (pt) 2003-08-05 2013-04-23 Novo Nordisk As Novos derivados de insulina
ES2574581T3 (es) 2003-08-14 2016-06-20 Novo Nordisk Health Care Ag Composición farmacéutica líquida acuosa de polipéptidos de tipo Factor VII
WO2005021022A2 (en) 2003-09-01 2005-03-10 Novo Nordisk A/S Stable formulations of peptides
ES2229931B1 (es) 2003-10-03 2006-01-16 Grifols, S.A. Composicion liquida bilogicamente estable de fviii, de fvw o del complejo fviii/fvw humanos.
ES2383691T3 (es) 2003-12-23 2012-06-25 Pharmacia Corporation Formulación líquida estable de hormonas de crecimiento
PL1740154T3 (pl) 2004-03-12 2009-11-30 Biodel Inc Kompozycje insuliny o ulepszonym wchłanianiu
WO2005117948A1 (en) 2004-06-01 2005-12-15 Ares Trading S.A. Method of stabilizing proteins
WO2006008238A1 (en) 2004-07-16 2006-01-26 Novo Nordisk A/S Method for selective acylation
DE602005022895D1 (de) 2004-08-12 2010-09-23 Schering Corp Stabile pegylierte interferon-formulierung
PT1778723E (pt) 2004-08-17 2013-01-25 Regeneron Pharma Formulações de antagonista de il-1
ES2575984T3 (es) 2004-11-12 2016-07-04 Novo Nordisk A/S Formulaciones estables de péptidos que contienen un análogo de GLP-1 acilado y una insulina basal
JP4874989B2 (ja) 2004-11-22 2012-02-15 ノヴォ ノルディスク アー／エス 可溶性で安定なインスリン含有調合物
EP1838290A2 (en) 2005-01-21 2007-10-03 Alza Corporation Therapeutic peptide formulations for coating microneedles with improved stability containing at least one counterion
US20080171695A1 (en) 2005-02-02 2008-07-17 Novo Nordisk A/S Insulin Derivatives
US20070086952A1 (en) 2005-09-29 2007-04-19 Biodel, Inc. Rapid Acting and Prolonged Acting Inhalable Insulin Preparations
CA2626357A1 (en) 2005-10-20 2007-04-26 Nastech Pharmaceutical Company Inc. Intranasal administration of rapid acting insulin
EP2505593A1 (en) 2005-12-28 2012-10-03 Novo Nordisk A/S Compositions comprising an acylated insulin and zinc and method of making the said compositions
MX2008013165A (es) 2006-04-12 2009-01-29 Biodel Inc Formulaciones de combinacion de insulina de accion rapida y accion larga.
CN101454019A (zh) 2006-04-12 2009-06-10 百达尔公司 速效和长效胰岛素联合制剂
US8796205B2 (en) 2006-05-09 2014-08-05 Novo Nordisk A/S Insulin derivative
WO2007135117A2 (en) 2006-05-24 2007-11-29 Novo Nordisk A/S Soluble, stable insulin-containing formulations
KR101699370B1 (ko) 2006-09-22 2017-02-14 노보 노르디스크 에이/에스 프로테아제 내성 인슐린 유사체
ES2526924T3 (es) 2007-08-15 2015-01-16 Novo Nordisk A/S Insulinas con una fracción acilo que comprende unidades repetitivas de aminoácidos que contienen alquilenglicol
EP2036539A1 (en) 2007-09-11 2009-03-18 Novo Nordisk A/S Stable formulations of amylin and its analogues
US8710000B2 (en) 2007-11-08 2014-04-29 Novo Nordisk A/S Insulin derivative
WO2009115469A1 (en) 2008-03-18 2009-09-24 Novo Nordisk A/S Protease stabilized, acylated insulin analogues
TWI451876B (zh) 2008-06-13 2014-09-11 Lilly Co Eli 聚乙二醇化之離脯胰島素化合物
ES2607003T3 (es) 2008-10-30 2017-03-28 Novo Nordisk A/S Tratamiento de diabetes mellitus utilizando inyecciones de insulina con una frecuencia de inyección inferior a la diaria
WO2011141407A1 (en) 2010-05-10 2011-11-17 Novo Nordisk A/S Process for the preparation of insulin-zinc complexes
AU2011202239C1 (en) 2010-05-19 2017-03-16 Sanofi Long-acting formulations of insulins
EP2389945A1 (en) 2010-05-28 2011-11-30 Sanofi-Aventis Deutschland GmbH Pharmaceutical composition comprising AVE0010 and insulin glargine
PL2632478T3 (pl) 2010-10-27 2020-03-31 Novo Nordisk A/S Leczenie cukrzycy z zastosowaniem zastrzyków insuliny podawanych w różnych odstępach czasu
RU2013123515A (ru) 2010-10-27 2014-12-10 Ново Нордиск А/С Лечение сахарного диабета с помощью инъекций инсулина, вводимых с различными интервалами
AU2012213432B2 (en) 2011-02-01 2016-10-13 Novo Nordisk A/S Purification of insulin
MX344925B (es) 2011-03-01 2017-01-11 N21 Acquisition Holding Llc Composiciones de insulina y cromo para el tratamiento y prevención de diabetes, hipoglucemia y alteraciones relacionadas.
EP2798546B1 (en) 2011-09-13 2020-12-23 Novo Nordisk A/S Adaptive system for optimizing a drug dosage regimen over time
WO2013153000A2 (en) 2012-04-11 2013-10-17 Novo Nordisk A/S Insulin formulations
JP6262206B2 (ja) 2012-05-01 2018-01-17 ノヴォ ノルディスク アー／エス 医薬組成物
JP6755175B2 (ja) 2013-03-20 2020-09-16 ノヴォ ノルディスク アー／エス インスリン投薬レジメン
US9839579B2 (en) 2013-04-24 2017-12-12 Corning Incorporated Delamination resistant pharmaceutical glass containers containing active pharmaceutical ingredients
US10137172B2 (en) 2013-04-30 2018-11-27 Novo Nordisk A/S Administration regime
EP3668892A1 (en) 2017-08-17 2020-06-24 Novo Nordisk A/S Novel acylated insulin analogues and uses thereof

Patent Citations (41)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3528960A (en) * 1968-10-07 1970-09-15 Lilly Co Eli N-carboxyaroyl insulins
US3868358A (en) * 1971-04-30 1975-02-25 Lilly Co Eli Protamine-insulin product
US4476118A (en) * 1982-02-05 1984-10-09 Novo Industri A/S Stabilized insulin preparations
US5177058A (en) * 1988-08-13 1993-01-05 Hoechst Aktiengesellschaft Pharmaceutical formulation for the treatment of diabetes mellitus
US5382574A (en) * 1989-12-21 1995-01-17 Novo Nordisk A/S Insulin preparations containing nicotinic acid or nicotinamide
US6011007A (en) * 1993-09-17 2000-01-04 Novo Nordisk A/S Acylated insulin
US5750497A (en) * 1993-09-17 1998-05-12 Novo Nordisk A/S Acylated insulin
US6869930B1 (en) * 1993-09-17 2005-03-22 Novo Nordisk A/S Acylated insulin
US5646242A (en) * 1994-11-17 1997-07-08 Eli Lilly And Company Selective acylation of epsilon-amino groups
US5830999A (en) * 1995-01-26 1998-11-03 Regents Of The University Of California Stabilization of insulin through ligand binding interations
US6620780B2 (en) * 1995-03-17 2003-09-16 Novo Nordisk A/S Insulin derivatives
US6251856B1 (en) * 1995-03-17 2001-06-26 Novo Nordisk A/S Insulin derivatives
US20020045731A1 (en) * 1996-02-21 2002-04-18 Novo Nordisk Of North America, Inc. Peptide derivatives
US6451970B1 (en) * 1996-02-21 2002-09-17 Novo Nordisk A/S Peptide derivatives
US5866538A (en) * 1996-06-20 1999-02-02 Novo Nordisk A/S Insulin preparations containing NaCl
US5905140A (en) * 1996-07-11 1999-05-18 Novo Nordisk A/S, Novo Alle Selective acylation method
US6504005B1 (en) * 1996-08-07 2003-01-07 Yeda Research And Development Co. Ltd. Long-acting drugs and pharmaceutical compositions comprising them
US5898067A (en) * 1997-02-07 1999-04-27 Novo Nordisk A/S Crystallization of proteins
US6451762B1 (en) * 1997-10-24 2002-09-17 Novo Nordisk A/S Aggregates of human insulin derivatives
US20020155994A1 (en) * 1997-10-24 2002-10-24 Svend Havelund Aggregates of human insulin derivatives
US20050222006A1 (en) * 1997-10-24 2005-10-06 Svend Havelund Aggregates of human insulin derivatives
US6335316B1 (en) * 1997-10-31 2002-01-01 Eli Lilly And Company Method for administering acylated insulin
US6531448B1 (en) * 1997-12-23 2003-03-11 Eli Lilly And Company Insoluble compositions for controlling blood glucose
US6174856B1 (en) * 1998-01-09 2001-01-16 Novo Nordisk A/S Stabilized insulin compositions
US6211144B1 (en) * 1998-10-16 2001-04-03 Novo Nordisk A/S Stable concentrated insulin preparations for pulmonary delivery
US6652886B2 (en) * 2001-02-16 2003-11-25 Expression Genetics Biodegradable cationic copolymers of poly (alkylenimine) and poly (ethylene glycol) for the delivery of bioactive agents
US20030004096A1 (en) * 2001-03-23 2003-01-02 Peter Boderke Zinc-free and low-zinc insulin preparations having improved stability
US8003605B2 (en) * 2002-03-13 2011-08-23 Novo Nordisk A/S Minimising body weight gain in insulin treatment
US20040006000A1 (en) * 2002-05-07 2004-01-08 Liselotte Langkjaer Novel formulations
US20040138099A1 (en) * 2002-11-29 2004-07-15 Draeger Eberhard Kurt Insulin administration regimens for the treatment of subjects with diabetes
US7544656B2 (en) * 2003-03-04 2009-06-09 The Technology Development Company, Ltd. Long acting injectable insulin composition and methods of making and using thereof
US20100009899A1 (en) * 2003-08-05 2010-01-14 Novo Nordisk A/S Novel Insulin Derivatives
US7615532B2 (en) * 2003-08-05 2009-11-10 Novo Nordisk A/S Insulin derivatives
US20060183668A1 (en) * 2003-08-05 2006-08-17 Novo Nordisk A/S Novel insulin derivatives
US7402565B2 (en) * 2003-11-14 2008-07-22 Novo Nordisk A/S Processes for making acylated insulin
US20080076705A1 (en) * 2005-02-02 2008-03-27 Kodra Janos T Insulin Derivatives
US8067362B2 (en) * 2005-02-02 2011-11-29 Novo Nordisk As Insulin derivatives
US20090137454A1 (en) * 2006-02-27 2009-05-28 Novo Nordisk A/S Insulin Derivatives
US20090239785A1 (en) * 2006-05-09 2009-09-24 Novo Nordisk A/S Insulin Derivative
US20100167990A1 (en) * 2007-06-13 2010-07-01 Novo Nordisk A/S Pharmaceutical Formulations Comprising an Insulin Derivative
US20110152185A1 (en) * 2007-11-16 2011-06-23 Novo Nordisk A/S Pharmaceutical Compositions Comprising GLP-1 Peptides or Extendin-4 and a Basal Insulin Peptide

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090011976A1 (en) * 2004-11-12 2009-01-08 Novo Nordisk A/S Stable Formulations Of Peptides
USRE46170E1 (en) 2005-02-02 2016-10-04 Novo Nordisk A/S Insulin derivatives
US8476228B2 (en) 2005-02-02 2013-07-02 Novo Nordisk A/S Insulin derivatives
US11167035B2 (en) 2005-12-28 2021-11-09 Novo Nordisk A/S Insulin compositions and method of making a composition
US20090137454A1 (en) * 2006-02-27 2009-05-28 Novo Nordisk A/S Insulin Derivatives
US8722620B2 (en) 2006-02-27 2014-05-13 Novo Nordisk A/S Insulin derivatives
US20090105121A1 (en) * 2006-05-09 2009-04-23 Ib Jonassen Insulin Derivative
US8796205B2 (en) 2006-05-09 2014-08-05 Novo Nordisk A/S Insulin derivative
US20100279931A1 (en) * 2007-11-08 2010-11-04 Novo Nordisk A/S Insulin Derivative
US8710000B2 (en) * 2007-11-08 2014-04-29 Novo Nordisk A/S Insulin derivative
US10596229B2 (en) 2010-10-27 2020-03-24 Novo Nordisk A/S Method of treating diabetes mellitus by administration, at specifically defined intervals, of a derivative of a naturally occurring insulin or insulin analogue, the derivative having a prolonged profile of action
US10137172B2 (en) 2013-04-30 2018-11-27 Novo Nordisk A/S Administration regime
US10040839B2 (en) 2014-02-28 2018-08-07 Novo Nordisk A/S Insulin derivatives and the medical uses hereof
US10596231B2 (en) 2016-12-16 2020-03-24 Novo Nordisk A/S Insulin containing pharmaceutical compositions
US10335464B1 (en) 2018-06-26 2019-07-02 Novo Nordisk A/S Device for titrating basal insulin

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