EP2451471A1 - Slow-acting insulin preparations - Google Patents

Slow-acting insulin preparations

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Publication number
EP2451471A1
EP2451471A1 EP10726993A EP10726993A EP2451471A1 EP 2451471 A1 EP2451471 A1 EP 2451471A1 EP 10726993 A EP10726993 A EP 10726993A EP 10726993 A EP10726993 A EP 10726993A EP 2451471 A1 EP2451471 A1 EP 2451471A1
Authority
EP
European Patent Office
Prior art keywords
arg
pro
asp
lys
glu
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP10726993A
Other languages
German (de)
French (fr)
Inventor
Isabell Schoettle
Christiane Fuerst
Verena Siefke-Henzler
Gerrit Hauck
Walter Kamm
Julia Schnieders
Jutta Carls
Gert Wolf
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Sanofi Aventis Deutschland GmbH
Original Assignee
Sanofi Aventis Deutschland GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Sanofi Aventis Deutschland GmbH filed Critical Sanofi Aventis Deutschland GmbH
Publication of EP2451471A1 publication Critical patent/EP2451471A1/en
Withdrawn legal-status Critical Current

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Classifications

    • 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
    • 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
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
    • A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the invention relates to an aqueous pharmaceutical formulation with a
  • Type II diabetes is not generally deficient in insulin, but in a large number of cases, especially in advanced stages, treatment with insulin may be indicated
  • the replacement of the body's insulin secretion by exogenous, usually subcutaneous administration of insulin generally does not approach the above-described quality of the physiological regulation of blood glucose.
  • derailments of blood glucose go up or down, which in their most severe forms can be life-threatening could be.
  • increased blood glucose levels without initial symptoms represent a considerable health risk over years.
  • the large-scale DCCT study in the USA The Diabetes Control and Compounds Trial Research Group (1993) N. Engl. J. Med. 329, 977-986) showed clearly that chronically elevated blood glucose levels are significantly responsible for the development of late diabetic damage. Diabetic late damage is microvascular and macrovascular
  • Insulin preparations are achieved. Fast-acting formulations are given at meal times to compensate for the postprandial increase in blood glucose. Slow-acting basal insulins should ensure the basic supply of insulin, especially at night, without leading to hypoglycaemia.
  • Insulin is a 51 amino acid polypeptide distributed among 2 amino acid chains: the 21 amino acid A chain and the 30 amino acid B chain. The chains are linked by 2 disulfide bridges. Insulin preparations have been used for diabetes therapy for many years. Not only naturally occurring insulins are used, but more recently also insulin derivatives and analogues.
  • Insulin analogs are analogues of naturally occurring insulins, viz
  • Human insulin or animal insulins which differ in substitution of at least one naturally occurring amino acid residue with other amino acids and / or addition / removal of at least one amino acid residue from the corresponding otherwise identical naturally occurring insulin. These may also be amino acids that are not naturally occurring.
  • Insulin derivatives are derivatives of naturally occurring insulin or a
  • Insulin analog obtained by chemical modification.
  • the chemical modification may e.g. in the addition of one or more particular chemical groups to one or more amino acids.
  • insulin derivatives and insulin analogues have a slightly altered effect on human insulin.
  • EP 0 375 437 and EP 0 678 522 EP 0 124 826 relates inter alia. on substitutions of B27 and B28.
  • EP 0 678 522 describes insulin analogues which have different amino acids in position B29, preferably proline, but not glutamic acid.
  • EP 0 375 437 comprises insulin analogues with lysine or arginine in B28, which may optionally be additionally modified in B3 and / or A21.
  • Protected modifications are modified in the asparagine in B3 and at least one other amino acid in positions A5, A15, A18 or A21.
  • insulin derivatives and insulin analogues have a slightly altered effect on human insulin.
  • insulin analogs are described in which at least one amino acid of positions B1-B6 is replaced by lysine or arginine. Such insulins have a prolonged action according to WO 92/00321.
  • the insulin analogues described in EP-A 0 368 187 also have a delayed action.
  • the concept of intensified insulin therapy seeks to reduce the health risk by aiming for a stable control of blood sugar levels by early administration of basal insulin.
  • the insulin preparations on the market of naturally occurring insulin for insulin substitution differ in the source of insulin (e.g., beef, pork, human insulin) and the composition with which the profile of action (onset and duration of action) can be affected.
  • Insulins include insulin glargine (Gly (A21) Arg (B31) Arg (B32) human insulin) with a prolonged duration of action. Insulin glargine is injected as an acidic, clear solution and due to its solubility properties it falls in the physiological pH range of the
  • Insulin glargine is injected once daily and is distinguished from other long-acting insulins by its low serum profile and the associated reduction in the risk of nocturnal hypoglycemia (Schubert-Zsilavecz et al., 2: 125-130 (2001)).
  • the specific preparation of insulin glargine, which leads to the prolonged duration of action, is in
  • the B chain end consists of an amidated basic amino acid residue such as lysine or argininamide, i. in the amidated basic amino acid residue at the B chain end, the carboxyl group of the terminal amino acid is in its amidated form, and
  • the N-terminal amino acid residue of the insulin A chain is a lysine or arginine residue
  • the amino acid position A21 is occupied by a glycine residue
  • An object of the invention is therefore an aqueous pharmaceutical
  • a further subject of the invention is a pharmaceutical formulation as described above, in which the insulin analogue is selected from a group comprising:
  • Another object of the invention is a pharmaceutical formulation as described above, wherein the isotonizing agent is selected from a group containing mannitol, sorbitol, lactose, dextrose, trehalose, sodium chloride, glycerol.
  • Another object of the invention is a pharmaceutical formulation as described above, having a pH in the range of pH 2.5 to 4.5, preferably in the range of pH 3.0 to 4.0, particularly preferably in the range of pH. 3 , 75th
  • Another object of the invention is a pharmaceutical formulation as described above, wherein the insulin, the insulin analogue and / or the insulin derivative is present in a concentration of 60-6000 nmol / ml
  • a further subject of the invention is a pharmaceutical formulation as described above in which glycerol is present in a concentration of 20 to 30 mg / ml, preferably in a concentration of 25 mg / ml.
  • a further subject of the invention is a pharmaceutical formulation as described above in which m-cresol is present in a concentration of 1 to 3 mg / ml, preferably in a concentration of 2 mg / ml.
  • Another object of the invention is a pharmaceutical formulation as described above in which zinc is present in a concentration of 0.01 or 0.03 or 0.08 mg / ml.
  • a further subject of the invention is a pharmaceutical formulation as described above, which additionally contains a glucagon-like peptide-1 (GLP1) or an analogue or derivative thereof, or exendin-3 or -4 or an analogue or derivative thereof ,
  • GLP1 glucagon-like peptide-1
  • Another object of the invention is a pharmaceutical formulation as described above, which additionally contains exendin-4.
  • Another object of the invention is a pharmaceutical formulation as described above in which an analog of exendin-4 is selected from a group containing
  • Another object of the invention is a pharmaceutical formulation as described above in which an analog of exendin-4 is selected from a group containing
  • Another object of the invention is a pharmaceutical formulation as described above, in which the peptide -LySo-NH 2 is added to the C-termini of the analogues of exendin-4.
  • Another object of the invention is a pharmaceutical formulation as described above in which an analog of exendin-4 is selected from a group containing
  • Another object of the invention is a pharmaceutical formulation as described above, in which in addition Arg 34 , Lys 26 (N ⁇ ( ⁇ -glutamyl (N ⁇ -hexadecanoyl))) GLP-1 (7-37) [liraglutide] or a pharmacologically tolerable salt thereof is included.
  • Another object of the invention is a pharmaceutical formulation as described above, in which the amino acid methionine is present, preferably in the concentration range of up to 10 mg / ml, more preferably of up to 3 mg / ml.
  • Another object of the invention is a method for producing a
  • Another object of the invention is a use of a formulation as described above for the treatment of diabetes mellitus.
  • Another object of the invention is a medicament for the treatment of
  • Diabetes mellitus consisting of a formulation as described above.
  • the preparation may further contain preservatives (e.g., phenol, cresol,
  • isotonizing agents e.g., mannitol, sorbitoi, lactose, dextrose,
  • Trehalose, sodium chloride, glycerol Buffer substances, salts, acids and alkalis and other excipients. These substances may be present individually or as mixtures.
  • Giycerol, dextrose, lactose, sorbitoi and mannitol are usually present in the pharmaceutical preparation in a concentration of 100-250 mM, NaCl in a concentration up to 150 mM.
  • Buffer substances e.g. Phosphate, acetate, citrate, arginine, glycylglycine or TRIS (i.e., 2-amino-2-hydroxymethyl-1, 3, -
  • Propandiol buffers and corresponding salts may be present in a concentration of 5-250 mM, preferably 10-100 mM.
  • Other excipients may include salts or arginine.
  • a further subject of the invention is a pharmaceutical formulation as described above in which the insulin analogue is in a concentration of 60-6000 nmol / ml (this corresponds approximately to a concentration of 0.35-70 mg / ml or 10-1000 Units / ml), preferably in a concentration of 240-3000 nmol / ml (this corresponds approximately to a concentration of 1.4-35 mg / ml or 40-500
  • the surfactant is present in a concentration of 5-200 ⁇ g / ml, preferably of 5-120 ⁇ g / ml and more preferably of 20-75 ⁇ g / ml.
  • Another object of the invention is a pharmaceutical formulation as stated above, in which glycerol and / or mannitol in a concentration of 100-250 mM, and / or NaCl preferably in a concentration up to 150 mM.
  • Another object of the invention is a pharmaceutical formulation as stated above, in which a buffer substance in a concentration of 5 - 250 mM is present.
  • Another object of the invention is a pharmaceutical insulin formulation, the further additives such. Contains salts which retard the release of insulin. Also mixtures of such sustained-release insulins described above
  • Formulations are included. Another object of the invention is a method for producing such pharmaceutical formulations. Likewise, another object of the invention is the use of such formulations for the treatment of diabetes mellitus.
  • Another object of the invention is the use or the addition of surfactants as a stabilizer during the production process of insulin,
  • Insulin analogues or insulin derivatives or their preparations are insulin analogues or insulin derivatives or their preparations.
  • Fig. 1 Hypoglycemic effect of new insulin analogues according to formula I in
  • Fig. 2 blood sugar lowering effect of new insulin analogues according to formula I in
  • Fig. 3 Hypoglycemic effect of YKL205 in the dog
  • Fig. 4 Zinc dependence of the hypoglycemic effect of YKL205 in the dog
  • the solution is prepared by adding about 25% water for injection.
  • One after the other are SAR161271 and the zinc chloride
  • the tonicity agent Glyceroi in the concentration 2.5% was selected.
  • the formulation is more stable compared to 0.8% NaCl as a tonicity agent.
  • precipitation was detected using NaCl during the preparation which was insoluble.
  • the osmolarity was 290 ⁇ 30 m ⁇ smol / kg for both substances.
  • m-cresol was chosen as a preservative. The concentration of 2 mg / ml was chosen, although 1.5 mg / ml would have been sufficient for preservation. Nevertheless, the higher m-cresol concentration was chosen due to microbiological safety aspects and specification specification. In addition, the formulations (except 2.1 mg / ml m-cresol) were placed on stability (3 months).
  • Example 4 Formulation of the amidated insulin derivatives
  • Examples 4 to 8 serve only to determine the biological, pharmacological and physicochemical properties of insulin analogues according to formula I by firstly providing formulations thereof (example 4) and then carrying out corresponding tests (examples 5 to 8).
  • a solution was prepared of the compounds as follows: The insulin analog according to the invention was dissolved at a target concentration of 240 ⁇ 5 ⁇ M in 1 mM hydrochloric acid with 80 ⁇ g / ml zinc (as zinc chloride).
  • compositions were used as solvent medium:
  • freeze-dried material was first about a 30% higher amount than due to the molecular weight and the desired
  • Target concentration of 240 ⁇ 5 ⁇ M the final solution was syringed with a 0.2 ⁇ m filter attachment into a with a septum and a crimp cap
  • Example 5 Evaluation of the hypoglycemic effect of new insulin analogues in the rat
  • the hypoglycemic effect of selected new insulin analogues is tested in male, healthy, normoglycemic Wistar rats.
  • Male rats are injected subcutaneously with a dose of 9 nmol / kg of an insulin analogue.
  • blood samples are taken from the animals and the blood sugar content is determined therein. The experiment clearly shows (see Fig. 1) that the insulin analog used according to the invention significantly delayed
  • Example 6 Evaluation of the hypoglycaemic effect of new insulin analogues in the dog
  • the hypoglycemic effect of selected new insulin analogues is tested in male, healthy, normoglycemic beagle dogs.
  • Male animals are injected subcutaneously with a dose of 6 nmol / kg of an insulin analogue.
  • blood samples are taken from the animals and the blood sugar content is determined therein.
  • the experiment clearly shows (see Fig. 2) that the insulin analog according to the invention used is a clear one
  • the hypoglycemic effect of selected new insulin analogues is tested in male, healthy, normoglycemic beagle dogs.
  • Male animals are injected subcutaneously with a dose of 6 nmol / kg and 12 nmol / kg of an insulin analogue.
  • blood samples are taken from the animals and the blood sugar content is determined therein.
  • Example 8 Evaluation of the hypoglycemic effect in dogs at different zinc concentrations in the formulation
  • FIG. 4 shows the result. Thereafter, the time-effect curve of the insulin analog according to the invention by the content of zinc ions in the formulation at the same concentration of insulin influence in such a way that at zero or low zinc content observed a rapid onset and the effect persists over 24 hours, while at higher zinc content low onset of action is observed and the insulin action persists well longer than 24 hours.
  • Example 9 The insulin analog according to the invention was dissolved at a target concentration of 240 + 5 ⁇ M in 1 mM hydrochloric acid with 80 ⁇ g / ml zinc (as zinc chloride).
  • a target concentration 240 + 5 ⁇ M in 1 mM hydrochloric acid with 80 ⁇ g / ml zinc (as zinc chloride).
  • the freeze-dried material was first weighed about 30% higher amount than required due to the molecular weight and the desired concentration. Thereafter, the present concentration was determined by means of analytical HPLC and the solution was then filled to the required volume to achieve the target concentration with 5 mM hydrochloric acid with 80 ug / mL zinc. If necessary, the pH was readjusted to 3.5 + 0.1.
  • the final solution was transferred via syringe with a 0.2 ⁇ m filter attachment to a sterile vial sealed with a septum and crimp cap.
  • the formulations eg with regard to an addition of isotonic agents, preservatives or buffer substances, was carried out.
  • Example 10 Evaluation of the hypoglycemic effect of novel insulin analogues in the rat
  • the hypoglycemic effect of selected new insulin analogs is tested in male, normal, normoglycemic Wistar rats.
  • Male rats are injected subcutaneously with a dose of 9 nmol / kg of an insulin analogue.
  • blood samples are taken from the animals and the blood sugar content is determined therein.
  • the experiment clearly shows (compare Fig. 4) that the insulin analog according to the invention leads to a significantly delayed onset of action and a longer, uniform duration of action.
  • Example 11 Evaluation of the hypoglycemic effect of new insulin analogues in the dog The hypoglycemic effect of selected new insulin analogues is tested in male, healthy, normoglycemic beagle dogs. Male animals are injected subcutaneously with a dose of 6 nmol / kg of an insulin analogue. Immediately before injecting the insulin analogue and at regular intervals up to forty-eight hours after the injection, blood samples are taken from the animals and the blood sugar content is determined therein. The experiment clearly shows that the insulin analog according to the invention leads to a markedly delayed, shallow onset of action and a longer, uniform duration of action.

Abstract

The invention relates to aqueous pharmaceutical formulations having an insulin analog, said formulations comprising 0.001 to 0.2 mg/ml of zinc, 0.1 to 5.0 mg/ml of a preservative, and 5.0 to 100 mg/ml of an isotonic agent, and the pH value of which is 5 or less. The invention further relates to the production thereof, the use thereof for treating diabetes mellitus, and a medication for treating diabetes mellitus.

Description

Beschreibung  description
Langsamwirkende Insulinzubereitungen Die Erfindung betrifft eine wässrige pharmazeutische Formulierungen mit einem Slow-acting insulin preparations The invention relates to an aqueous pharmaceutical formulation with a
Insulinanalogon, die Insulin analog, the
0,001 bis 0,2 mg/ml Zink, From 0.001 to 0.2 mg / ml of zinc,
0,1 bis 5,0 mg/ml eines Konservierungsmittels und  0.1 to 5.0 mg / ml of a preservative and
5,0 bis 100 mg/ml eines Isotonisierungsmittels enthält und Contains 5.0 to 100 mg / ml of an isotonizing agent and
deren pH-Wert 5 oder kleiner ist; sowie deren Herstellung, Verwendung zur whose pH is 5 or less; as well as their production, use for
Behandlung von Diabetes mellitus und ein Arzneimittel zur Behandlung von Diabetes Mellitus.  Treatment of diabetes mellitus and a drug for the treatment of diabetes mellitus.
Weltweit leidet eine zunehmende Zahl an Menschen an Diabetes mellitus. Darunter sind viele sogenannte Typ I-Diabetiker, für die die Substitution der fehlenden An increasing number of people worldwide suffer from diabetes mellitus. Among them are many so-called type I diabetics, for whom the substitution of the missing
endokrinen Insulinsekretion die einzige derzeit mögliche Therapie darstellt. Die endocrine insulin secretion represents the only therapy currently possible. The
Betroffenen sind lebenslang, in der Regel mehrmals täglich, auf Insulininjektionen angewiesen. Im Gegensatz zum Typ I-Diabetes besteht beim Typ Il-Diabetes nicht grundsätzlich ein Mangel an Insulin, jedoch wird in einer Vielzahl von Fällen, vor allem im fortgeschrittenen Stadium, die Behandlung mit Insulin, gegebenenfalls in Patients are lifelong, usually several times a day, dependent on insulin injections. In contrast to type I diabetes, type II diabetes is not generally deficient in insulin, but in a large number of cases, especially in advanced stages, treatment with insulin may be indicated
Kombination mit einem oralen Antidiabetikum, als günstigste Therapieform angesehen. Beim Gesunden ist die Insulinfreisetzung durch den Pankreas strikt an die  Combination with an oral antidiabetic, considered as the most beneficial form of therapy. In the healthy the insulin release by the pancreas is strictly to the
Konzentration der Blutglucose gekoppelt. Erhöhte Blutglucosespiegel, wie sie nach Mahlzeiten auftreten, werden durch eine entsprechende Steigerung der Concentration of blood glucose coupled. Increased blood glucose levels, as they occur after meals, are boosted by a corresponding increase
Insulinsekretion rasch kompensiert. Im nüchternen Zustand sinkt der Insulin secretion rapidly compensated. In the fasting state of the sinks
Plasmainsulinspiegel auf einen basalen Wert ab, der ausreicht, eine kontinuierliche Versorgung insulinsensitiver Organe und Gewebe mit Glucose zu gewährleisten und die hepatische Glucoseproduktion in der Nacht niedrig zu halten. Der Ersatz der körpereigenen Insulinsekretion durch exogene, meist subkutane Applikation von Insulin erreicht in der Regel die oben beschriebene Qualität der physiologischen Regulation der Blutglucose nicht annähernd. Häufig kommt es zu Entgleisungen der Blutglucose nach oben oder unten, die in ihren schwersten Formen lebensbedrohlich sein können. Daneben stellen jedoch auch über Jahre erhöhte Blutglucosespiegel ohne anfängliche Symptome ein erhebliches Gesundheitsrisiko dar. Die großangelegte DCCT-Studie in den USA (The Diabetes Control and Compiications Trial Research Group (1993) N. Engl. J. Med. 329, 977-986) wies eindeutig nach, daß chronisch erhöhte Blutglucosespiegel wesentlich für die Entwicklung diabetischer Spätschäden verantwortlich sind. Diabetische Spätschäden sind mikro- und makrovaskuläre Plasma insulin levels to a basal level sufficient to ensure a continuous supply of insulin-sensitive organs and tissues with glucose and to keep the hepatic glucose production at night low. The replacement of the body's insulin secretion by exogenous, usually subcutaneous administration of insulin generally does not approach the above-described quality of the physiological regulation of blood glucose. Often, derailments of blood glucose go up or down, which in their most severe forms can be life-threatening could be. In addition, however, increased blood glucose levels without initial symptoms represent a considerable health risk over years. The large-scale DCCT study in the USA (The Diabetes Control and Compounds Trial Research Group (1993) N. Engl. J. Med. 329, 977-986) showed clearly that chronically elevated blood glucose levels are significantly responsible for the development of late diabetic damage. Diabetic late damage is microvascular and macrovascular
Schädigungen, die sich u.U. als Retino-, Nephro-, oder Neuropathie manifestieren und zu Erblindung, Nierenversagen sowie dem Verlust von Extremitäten führen und darüber hinaus mit einem erhöhten Risiko für Herz/Kreislauferkrankungen Damage that may u.U. manifest as retinopathy, nephropathy or neuropathy and lead to blindness, renal failure and loss of extremities and, moreover, an increased risk of cardiovascular disease
einhergehen. Daraus ist abzuleiten, daß eine verbesserte Therapie des Diabetes in erster Linie darauf abzielen muß, die Blutglucose möglichst eng im physiologischen Bereich zu halten. Nach dem Konzept der intensivierten Insulintherapie soll dies durch mehrmals tägliche Injektionen von schnell und langsam wirkenden accompanied. It can be deduced from this that an improved therapy of diabetes must primarily aim to keep the blood glucose as close as possible to the physiological range. According to the concept of intensified insulin therapy, this should be done by injecting fast and slow acting injections several times a day
Insulinzubereitungen erreicht werden. Rasch wirkende Formulierungen werden zu den Mahlzeiten gegeben, um den postprandialen Anstieg der Blutglucose auszugleichen. Langsam wirkende Basalinsuline sollen die Grundversorgung mit Insulin insbesondere während der Nacht sicherstellen, ohne zu einer Hypoglykämie zu führen. Insulin preparations are achieved. Fast-acting formulations are given at meal times to compensate for the postprandial increase in blood glucose. Slow-acting basal insulins should ensure the basic supply of insulin, especially at night, without leading to hypoglycaemia.
Insulin ist ein Polypeptid aus 51 Aminosäuren, die sich auf 2 Aminosäureketten verteilen: die A Kette mit 21 Aminosäuren und die B-Kette mit 30 Aminosäuren. Die Ketten sind durch 2 Disulfidbrücken miteinander verbunden. Insulinzubereitungen werden seit vielen Jahren zur Diabetestherapie eingesetzt. Dabei werden nicht nur natürlich vorkommende Insuline verwendet, sondern neuerdings auch Insulinderivate und -analoga. Insulin is a 51 amino acid polypeptide distributed among 2 amino acid chains: the 21 amino acid A chain and the 30 amino acid B chain. The chains are linked by 2 disulfide bridges. Insulin preparations have been used for diabetes therapy for many years. Not only naturally occurring insulins are used, but more recently also insulin derivatives and analogues.
Insulinanaloga sind Analoga von natürlich vorkommenden Insulinen, nämlich Insulin analogs are analogues of naturally occurring insulins, viz
Humaninsulin oder tierischen Insulinen, welche sich durch Substitution wenigstens eines natürlich auftretenden Aminosäurerestes mit anderen Aminosäuren und/oder Addition/Entfernen wenigstens eines Aminosäurerestes von dem entsprechenden, ansonsten gleichen natürlich vorkommenden Insulin unterscheiden. Es kann sich dabei auch um Aminosäuren handeln, die nicht natürlich vorkommen. Insulinderivate sind Derivate von natürlich vorkommendem Insulin oder einem Human insulin or animal insulins which differ in substitution of at least one naturally occurring amino acid residue with other amino acids and / or addition / removal of at least one amino acid residue from the corresponding otherwise identical naturally occurring insulin. These may also be amino acids that are not naturally occurring. Insulin derivatives are derivatives of naturally occurring insulin or a
Insulinanalogon, welche durch chemische Modifizierung erhalten werden. Die chemische Modifikation kann z.B. in der Addition einer oder mehrerer bestimmter chemischer Gruppen an eine oder mehrere Aminosäuren bestehen. In der Regel haben Insulinderivate und Insulinanaloga gegenüber humanem Insulin eine etwas veränderte Wirkung. Insulin analog obtained by chemical modification. The chemical modification may e.g. in the addition of one or more particular chemical groups to one or more amino acids. In general, insulin derivatives and insulin analogues have a slightly altered effect on human insulin.
Insulinanaloga mit beschleunigtem Wirkungseintritt werden in EP 0 214 826, Insulin analogs with accelerated onset of action are described in EP 0 214 826,
EP 0 375 437 und EP 0 678 522 beschrieben. EP 0 124 826 bezieht sich u.a. auf Substitutionen von B27 und B28. EP 0 678 522 beschreibt Insulinanaloga, die in der Position B29 verschiedene Aminosäuren, vorzugsweise Prolin, aufweisen, jedoch nicht Glutaminsäure. EP 0 375 437 and EP 0 678 522. EP 0 124 826 relates inter alia. on substitutions of B27 and B28. EP 0 678 522 describes insulin analogues which have different amino acids in position B29, preferably proline, but not glutamic acid.
EP 0 375 437 umfaßt Insulinanaloga mit Lysin oder Arginin in B28, die optional zusätzlich in B3 und/oder A21 modifiziert sein können.  EP 0 375 437 comprises insulin analogues with lysine or arginine in B28, which may optionally be additionally modified in B3 and / or A21.
In der EP 0 419 504 werden Insulinanaloga offenbart, die gegen chemische In EP 0 419 504 insulin analogs are disclosed which are resistant to chemical
Modifikationen geschützt sind, in dem Asparagin in B3 und wenigstens eine weitere Aminosäure in den Positionen A5, A15, A18 oder A21 verändert sind. In der Regel haben Insulinderivate und Insulinanaloga gegenüber humanem Insulin eine etwas veränderte Wirkung. Protected modifications are modified in the asparagine in B3 and at least one other amino acid in positions A5, A15, A18 or A21. In general, insulin derivatives and insulin analogues have a slightly altered effect on human insulin.
In der WO 92/00321 werden Insulinanaloga beschrieben, bei denen wenigstens eine Aminosäure der Positionen B1-B6 durch Lysin oder Arginin ersetzt ist. Derartige Insuline weisen gemäß WO 92/00321 eine verlängerte Wirkung auf. Eine verzögerte Wirkung weisen auch die in der EP-A 0 368 187 beschriebenen Insulinanaloga auf. Das Konzept der intensivierten Insulintherapie versucht das Gesundheitsrisiko abzumindern, indem eine stabile Kontrolle des Blutzuckerspiegels durch frühe Gabe von Basalinsulinen angestrebt wird. Ein Beispiel für ein gängiges Basalinsulin ist das Medikament Lantus® (Wirkstoff: Insulin Glargin = GIy (A21 ), Arg (B31 ), Arg (B32)In WO 92/00321 insulin analogs are described in which at least one amino acid of positions B1-B6 is replaced by lysine or arginine. Such insulins have a prolonged action according to WO 92/00321. The insulin analogues described in EP-A 0 368 187 also have a delayed action. The concept of intensified insulin therapy seeks to reduce the health risk by aiming for a stable control of blood sugar levels by early administration of basal insulin. An example of a common basal insulin is the drug Lantus ® (active ingredient: insulin glargine = Gly (A21), Arg (B31), Arg (B32)
Humaninsulin). Generell gilt es, bei der Entwicklung neuer, verbesserter Basalinsuline die Zahl hypoglykämischer Ereignisse zu minimieren. Ein ideales Basalinsulin wirkt dabei sicher in jedem Patienten mindestens 24 Stunden. Idealerweise setzt die Human insulin). In general, it is important to minimize the number of hypoglycemic events in the development of new, improved basal insulins. An ideal basal insulin works while sure in every patient at least 24 hours. Ideally, the
Insulinwirkung verzögert und mit einem möglichst flachen Zeit- / Wirkungsprofil ein, so dass die Gefahr einer kurzfristigen Unterzuckerung deutlich minimiert ist und die Applikation sogar ohne vorherige Einnahme von Nahrungsmitteln erfolgen kann. Eine gute Versorgung mit Basalinsulin ist dann gegeben, wenn die Insulinwirkung möglichst lange gleichbleibend anhält, d.h. der Körper mit einer konstanten Menge Insulin versorgt wird. Damit ist die Gefahr hypoglykämischer Ereignisse gering und eine patienten- und tagesspezifische Variabilität minimiert. Das pharmakokinetische Profil eines idealen Basalinsulins sollte also durch einen verzögerten Wirkeintritt und durch eine verzögerte, d.h. lang anhaltende und gleichmäßige Wirkung gekennzeichnet sein. Delayed insulin action and with a flat as possible time / effect profile, so that the risk of short-term hypoglycemia is significantly minimized and the application can take place even without prior ingestion of food. A good supply of basal insulin is given when the insulin effect persists as long as possible, ie. the body is supplied with a constant amount of insulin. This minimizes the risk of hypoglycemic events and minimizes patient and tag-specific variability. The pharmacokinetic profile of an ideal basal insulin should therefore be characterized by a delayed onset of action and a delayed, i.e. long-lasting and uniform effect.
Die auf dem Markt befindlichen Insulinzubereitungen von natürlich vorkommenden Insulinen zur Insulinsubstitution unterscheiden sich in der Herkunft des Insulins (z.B. Rind, Schwein, Humaninsulin), sowie der Zusammensetzung, womit das Wirkprofil (Wirkeintritt und Wirkdauer) beeinflusst werden kann. Durch Kombination The insulin preparations on the market of naturally occurring insulin for insulin substitution differ in the source of insulin (e.g., beef, pork, human insulin) and the composition with which the profile of action (onset and duration of action) can be affected. By combination
verschiedener Insulinpräparate lassen sich unterschiedlichste Wirkprofile erzielen und möglichst physiologische Blutzuckerwerte einstellen. Die rekombinante DNA Various insulin preparations can be used to achieve a wide variety of activity profiles and to adjust physiological blood sugar levels as far as possible. The recombinant DNA
Technologie ermöglicht heutzutage die Herstellung von solchen modifizierten Technology today makes it possible to produce such modified ones
Insulinen. Hierzu zählt Insulin Glargin (Gly(A21 )-Arg(B31 )-Arg(B32)-Humaninsulin) mit einer verlängerten Wirkdauer. Insulin Glargin wird als saure, klare Lösung injiziert und fällt aufgrund seiner Lösungseigenschaften im physiologischen pH-Bereich des Insulins. These include insulin glargine (Gly (A21) Arg (B31) Arg (B32) human insulin) with a prolonged duration of action. Insulin glargine is injected as an acidic, clear solution and due to its solubility properties it falls in the physiological pH range of the
Subkutangewebes als stabiles Hexamerassoziat aus. Insulin Glargin wird einmal täglich injiziert und zeichnet sich gegenüber anderen langwirksamen Insulinen durch sein flaches Serumprofil und die damit verbundene Reduktion der Gefahr nächtlicher Hypoglykämien aus (Schubert-Zsilavecz et al., 2:125-130(2001 )). Die spezifische Zubereitung des Insulin Glargins, die zur verlängerten Wirkdauer führt, ist im Subcutaneous tissue as a stable hexamer associate. Insulin glargine is injected once daily and is distinguished from other long-acting insulins by its low serum profile and the associated reduction in the risk of nocturnal hypoglycemia (Schubert-Zsilavecz et al., 2: 125-130 (2001)). The specific preparation of insulin glargine, which leads to the prolonged duration of action, is in
Gegensatz zu bisher beschriebenen Zubereitungen durch einen klare Lösung mit saurem pH-Wert gekennzeichnet. Gerade bei saurem pH-Wert zeigen Insuline jedoch eine verringerte Stabilität und eine erhöhte Aggregationsneigung bei thermischer und physikalisch-mechanischer Belastung, die sich in Form von Trübungen und Contrary to previously described preparations characterized by a clear solution with acidic pH. However, especially at acidic pH insulins show a reduced stability and an increased tendency to aggregate under thermal and physical-mechanical stress, resulting in turbidity and
Ausfällungen (Partikelbildungen) bemerkbar machen kann (Brange et al., J. Ph. Sei 86:517-525(1997)). Der vorliegenden Erfindung lag somit die Aufgabe zugrunde, weitere Formulierungen für im Sauren lösliche Insulinanaloga zu finden, die einen verzögerten Wirkeintritt und eine längere Wirkdauer, d.h. einen extrem flachen und lang andauernden, Precipitations (particle formation) (Brange et al., J. Ph. Sei 86: 517-525 (1997)). It is an object of the present invention to find further formulations for acid-soluble insulin analogues which have a delayed onset of action and a longer duration of action, ie an extremely shallow and long lasting,
gleichmäßigen Wirkungsverlauf aufweisen. Damit wird die Gefahr von have a uniform course of action. This will increase the risk of
hypoglykämischen Ereignissen nochmals deutlich minimiert. Hypoglycemic events significantly minimized again.
Es wurde überraschend gefunden, dass solche Formulierungen zu dem It has surprisingly been found that such formulations to the
beschriebenen wünschenswerten basalen Zeit- / Wirkungsprofil führen, wenn die Insulinanaloga durch die Merkmale charakterisiert sind, dass described desirable basal time / activity profile, when the insulin analogues are characterized by the features that
• das B- Kettenende aus einem amidierten basischen Aminosäurerest wie Lysin bzw. Argininamid besteht, d.h. bei dem amidierten basischen Aminosäurerest am B- Kettenende liegt die Carboxylgruppe der endständigen Aminosäure in ihrer amidierten Form vor, und  • the B chain end consists of an amidated basic amino acid residue such as lysine or argininamide, i. in the amidated basic amino acid residue at the B chain end, the carboxyl group of the terminal amino acid is in its amidated form, and
• der N-terminale Aminosäurerest der Insulin A-Kette ein Lysin- oder Argininrest ist, und The N-terminal amino acid residue of the insulin A chain is a lysine or arginine residue, and
• die Aminosäureposition A8 durch einen Histidinrest besetzt wird, und  • the amino acid position A8 is occupied by a histidine residue, and
• die Aminosäureposition A21 durch einen Glycinrest besetzt wird, und  The amino acid position A21 is occupied by a glycine residue, and
• zwei Substitutionen neutraler Aminosäuren durch saure Aminosäuren, zwei  • two substitutions of neutral amino acids by acidic amino acids, two
Additionen negativ geladener Aminosäurereste oder je eine solche Substitution und eine solche Addition jeweils in den Positionen A5, A15, A18, B-1 , BO, B1 , B2, B3 undAdditions of negatively charged amino acid residues or such a substitution and such addition in the positions A5, A15, A18, B-1, BO, B1, B2, B3 and
B4 erfolgt sind; und B4 are done; and
0,001 bis 0,2 mg/ml Zink,  From 0.001 to 0.2 mg / ml of zinc,
0,1 bis 5,0 mg/ml eines Konservierungsmittels und  0.1 to 5.0 mg / ml of a preservative and
5,0 bis 100 mg/ml eines Isotonisierungsmittels enthalten sind und 5.0 to 100 mg / ml of an isotonizing agent are contained and
deren pH-Wert 5 oder kleiner ist. whose pH is 5 or less.
Ein Gegenstand der Erfindung ist daher eine wässrige pharmazeutische An object of the invention is therefore an aqueous pharmaceutical
Formulierungen mit einem Insulinanalogon der Formel I S S Formulations with an insulin analog of the formula I. SS
1 5 | 10j 15 20  1 5 | 10j 15 20
AO G I V E A5 C C H S I C S L Y A15 L E A18 Y C G  AO G I V E A5 C C H S I C S L Y A15 L E A18 Y C G
! (SEQ ID NO: 1) \ A-Kette ! (SEQ ID NO: 1) \ A chain
B-I BO Bl B2 B3 B4 H L C G S H L V E A L Y L V C G E R G F F Y B-I BO B1 B2 B3 B4 H L C G S H L V E A L Y L V C G E R G F F Y
1 5 10 15 20 25 1 5 10 15 20 25
T P B29 B30 B31 B32 (SEQ ID NO: 2) T P B29 B30 B31 B32 (SEQ ID NO: 2)
B-Kette B chain
30 wobei  30 where
AO Lys oder Arg; A5 Asp, GIn oder GIu; AO Lys or Arg; A5 Asp, GIn or GIu;
A15 Asp, GIu oder GIn; A15 Asp, Glu or GIn;
A18 Asp, GIu oder Asn; A18 Asp, Glu or Asn;
B-1 Asp, GIu oder eine Aminogruppe; B-1 Asp, Glu or an amino group;
BO Asp, GIu oder eine chemische Bindung; B1 Asp, GIu oder Phe; BO Asp, Glu or a chemical bond; B1 Asp, Glu or Phe;
B2 Asp, GIu oder VaI; B2 Asp, Glu or VaI;
B3 Asp, GIu oder Asn; B3 Asp, Glu or Asn;
B4 Asp, GIu oder GIn; B29 Lys oder einer chemischen Bindung; B4 Asp, Glu or GIn; B29 Lys or a chemical bond;
B30 Thr oder einer chemischen Bindung; B31 Arg, Lys oder einer chemischen Bindung; B30 Thr or a chemical bond; B31 Arg, Lys or a chemical bond;
B32 Arg-Amid, Lys-Amid oder einer Aminogruppe entspricht, wobei zwei Aminosäurereste der Gruppe enthaltend A5, A15, A18, B-1 , BO, B1 , B2, B3 und B4 gleichzeitig und unabhängig voneinander Asp oder GIu B32 Arg amide, Lys amide or an amino group, wherein two amino acid residues of the group containing A5, A15, A18, B-1, BO, B1, B2, B3 and B4 are simultaneously and independently Asp or GIu
entsprechen, oder ein pharmakologisch tolerierbares Salz davon; und die or a pharmacologically tolerable salt thereof; and the
0,001 bis 0,2 mg/ml Zink, From 0.001 to 0.2 mg / ml of zinc,
0,1 bis 5,0 mg/ml eines Konservierungsmittels und  0.1 to 5.0 mg / ml of a preservative and
5,0 bis 100 mg/ml eines Isotonisierungsmittels enthält und Contains 5.0 to 100 mg / ml of an isotonizing agent and
deren pH-Wert 5 oder kleiner ist. whose pH is 5 or less.
Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, bei der das Insulinanalogon ausgewählt ist aus einer Gruppe enthaltend: A further subject of the invention is a pharmaceutical formulation as described above, in which the insulin analogue is selected from a group comprising:
Arg (AO), His (A8), GIu (A5), Asp (A18), GIy (A21 ), Arg (B31), Arg (B32) - NH2 Humaninsuiin, Arg (AO), His (A8), GIu (A5), Asp (A18), GIy (A21 ), Arg (B31 ), Lys (B32) - NH2 Humaninsuiin, Arg (AO), His (A8), GIu (A15), Asp (A18), GIy (A21 ), Arg (B31 ), Arg (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIu (A15), Asp (A18), GIy (A21 ), Arg (B31 ), Lys (B32) - NH2 Humaninsulin, Arg (AO), His (A8), Glu(A5), GIu (A15), GIy (A21 ), Arg (B31 ), Arg (B32) - NH2 Humaninsulin, Arg (AO)1 His (A8), GIu (A5), GIu (A15), GIy (A21 ), Arg (B31 ), Lys (B32) - NH2 Humaninsulin, Arg (AO), His(A8), GIu (A5), GIy (A21 ), Asp (B3), Arg (B31), Arg (B32) - NH2 Humaninsu!in, Arg (AO), His(A8), GIu (A5), GIy (A21 ), Asp (B3), Arg (B31 ), Lys (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIu (A15), GIy (A21 ), Asp (B3), Arg (B31 ), Arg (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIu (A15), GIy (A21 ), Asp (B3), Arg (B31 ), Lys (B32) - NH2 Humaninsuiin, Arg (AO), His (A8), Asp (A18), GIy (A21 ), Asp (B3), Arg (B31 ), Arg (B32) - NH2 Humaninsuiin, Arg (AO), His (A8), Asp (A18), GIy (A21 ), Asp (B3), Arg (B31 ), Lys (B32) - NH2 Humaninsulin, Arg (AO), His(A8), GIy (A21 ), Asp (B3), GIu (B4), Arg (B31 ), Arg (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIy (A21), Asp (B3), GIu (B4), Arg (B31 ), Lys (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIu (A5), GIy (A21 ), GIu (B4), Arg (B31 ), Arg (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIu (A5), GIy (A21 ), GIu (B4), Arg (B31 ), Lys (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIu (A15), GIy (A21 ), GIu (B4), Arg (B31 ), Arg (B32) - NH2 Humaninsulin, Arg (AO)1 His (A8), GIu (A15), GIy (A21 ), Giu (B4), Arg (B31 ), Lys (B32) - NH2 Humaninsuiin, Arg (AO), His (A8), Asp (A18), GIy (A21), Giu (B4), Arg (B31 ), Arg (B32) - NH2 Humaninsuiin, Arg (AO), His (A8), Asp (A18), GIy (A21), Giu (B4), Arg (B31 ), Lys (B32) - NH2 Humaninsuiin, Arg (AO), His (A8), GIu (A5), GIy (A21 ), GIu (BO)1 Arg (B31 ), Arg (B32) - NH2 Humaninsulin, Arg (AO)1 His (A8), GIu (A5), GIy (A21 ), GIu (BO)1 Arg (B31 ), Lys (B32) - NH2 Humaninsulin, Arg (AO)1 His (A8), GIu (A15), GIy (A21 ), Giu (BO), Arg (B31 ), Arg (B32) - NH2 Humaninsuiin, Arg (AO)1 His (A8), GIu (A15), GIy (A21 ), GIu (BO), Arg (B31 ), Lys (B32) - NH2 Humaninsulin, Arg (AO), His (A8), Asp (A18), GIy (A21), GIu (BO), Arg (B31 ), Arg (B32) - NH2 Humaninsulin, Arg (AO)1 His (A8), Asp (A18) ,GIy (A21 ), Giu (BO)1 Arg (B31 ), Lys (B32) - NH2 Humaninsuiin, Arg (AO), His (A8), GIu (A5), GIy (A21), Asp (B1 ), Arg (B31 ), Arg (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIu (A5), GIy (A21), Asp (B1 ), Arg (B31 ), Lys (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIu (A15), GIy (A21 ), Asp (B1 ), Arg (B31 ), Arg(B32) - NH2 Humaninsulin, Arg (AO)1 His (A8), GIu (A15), GIy (A21 ), Asp (B1 ), Arg (B31 ), Lys (B32) - NH2 Humaninsulin, Arg (AO)1 His (A8), Asp (A18), GIy (A21 ), Asp (B1 ), Arg (B31 ), Arg (B32) - NH2 Humaninsuiin, Arg (AO)1 His (A8), Asp (A18), GIy (A21 ), Asp (B1), Arg (B31 ), Lys (B32) - NH2 Humaninsuiin, Arg (AO)1 His (A8), GIy (A21 ), GIu (BO)1 Asp (B1 ), Arg (B31 ), Arg (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIy (A21 ), GIu (BO)1 Asp (B1 ), Arg (B31 ), Lys (B32) - NH2 Humaninsulin, Arg (AO), His (A8), Asp (A18), GIy (A21 ), Asp (B3), Arg (B30), Arg (B31) - NH2 Humaninsuiin, Arg (AO), His (A8), Asp (A18), GIy (A21 ), Asp (B3), Arg (B30), Lys (B31) - NH2 Humaninsuiin. Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, wobei das Konservierungsmittel ausgewählt wird aus einer Gruppe enthaltend Phenol, m-Cresol, Chlorkresoi, Benzylalkohoi, Parabene. Arg (AO), His (A8), Glu (A5), Asp (A18), Gly (A21), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5), Asp (A18), Gly (A21), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A15), Asp (A18), Gly ( A21), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A15), Asp (A18), Gly (A21), Arg (B31), Lys (B32 ) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5), Glu (A15), Gly (A21), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO) 1 His (A8), Glu (A5), Glu (A15), Gly (A21), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5), Gly (A21), Asp (B3), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5), Gly (A21), Asp (B3), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A15), Gly (A21), Asp (B3), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A15), Gly (A21), Asp (B3), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8 ), Asp (A18), Gly (A21), A sp (B3), Arg (B31), Arg (B32) - NH 2 Humaninsuiin, Arg (AO), His (A8), Asp (A18), Gly (A21), Asp (B3), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Gly (A21), Asp (B3), Glu (B4), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Gly (A21), Asp (B3), Glu (B4), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5), Gly (A21), Glu (B4), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5), Gly (A21), Glu ( B4), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A15), Gly (A21), Glu (B4), Arg (B31), Arg (B32 ) - NH 2 human insulin, Arg (AO) 1 His (A8), Glu (A15), Gly (A21), Giu (B4), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Asp (A18), Gly (A21), Giu (B4), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Asp (A18), Gly (A21), Giu (B4), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5), Gly (A21), Glu (BO) 1 Arg ( B31), Arg (B32) - NH 2 human insulin, Arg (AO) 1 His (A8), Glu (A5), Gly (A21), Glu (BO) 1 Arg (B31), Lys (B32) - NH 2 human insulin , Arg (AO) 1 His (A8), Glu (A15), Gly (A21), Giu (BO), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO) 1 His (A8), GIu (A15), GIy (A21), Glu (BO), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Asp (A18), Gly (A21), Glu (BO), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO) 1 His (A8), Asp (A18), Gly (A21), Giu (BO) 1 Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO ), His (A8), Glu (A5), Gly (A21), Asp (B1), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5) , Gly (A21), Asp (B1), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A15), Gly (A21), Asp (B1), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO) 1 His (A8), Glu (A15), Gly (A21), Asp (B1), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO) 1 His (A8), Asp (A18), Gly (A21), Asp (B1), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO) 1 His (A8 ), Asp (A18), Gly (A21), Asp (B1), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO) 1 His (A8), Gly (A21), Glu (BO) 1 Asp (B1), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Gly (A21), Glu (BO) 1 Asp (B1), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Asp (A18), Gly (A21), Asp (B3), Arg (B30), Arg (B31) - NH 2 human insulin, Arg (AO), His (A8), Asp (A18), Gly (A21), Asp (B3), Arg (B30), Lys (B31) - NH 2 Humaninsuiin. Another object of the invention is a pharmaceutical formulation as described above, wherein the preservative is selected from a group containing phenol, m-cresol, Chlorkresoi, Benzylalkohoi, parabens.
Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, wobei das Isotonisierungsmittel ausgewählt wird aus einer Gruppe enthaltend Mannitol, Sorbitol, Lactose, Dextrose, Trehalose, Natriumchlorid, Glycerol. Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, mit einem pH-Wert im Bereich von pH 2,5 - 4,5, bevorzugt im Bereich von pH 3,0 - 4,0, besonders bevorzugt im Bereich von pH 3,75. Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, wobei das Insulin, das Insulinanalogon und/oder das Insulinderivat in einer Konzentration von 60 - 6000 nmol/ml vorliegt Another object of the invention is a pharmaceutical formulation as described above, wherein the isotonizing agent is selected from a group containing mannitol, sorbitol, lactose, dextrose, trehalose, sodium chloride, glycerol. Another object of the invention is a pharmaceutical formulation as described above, having a pH in the range of pH 2.5 to 4.5, preferably in the range of pH 3.0 to 4.0, particularly preferably in the range of pH. 3 , 75th Another object of the invention is a pharmaceutical formulation as described above, wherein the insulin, the insulin analogue and / or the insulin derivative is present in a concentration of 60-6000 nmol / ml
Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, bei der Glycerol in einer Konzentration von 20 bis 30 mg/ml, bevorzugt in einer Konzentration von 25 mg/ml vorliegt. A further subject of the invention is a pharmaceutical formulation as described above in which glycerol is present in a concentration of 20 to 30 mg / ml, preferably in a concentration of 25 mg / ml.
Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, bei der m-Cresol in einer Konzentration von 1 bis 3 mg/ml, bevorzugt in einer Konzentration von 2 mg/ml vorliegt. A further subject of the invention is a pharmaceutical formulation as described above in which m-cresol is present in a concentration of 1 to 3 mg / ml, preferably in a concentration of 2 mg / ml.
Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, bei der Zink in einer Konzentration 0,01 oder 0,03 oder 0,08 mg/ml vorliegt. Another object of the invention is a pharmaceutical formulation as described above in which zinc is present in a concentration of 0.01 or 0.03 or 0.08 mg / ml.
Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, bei der noch zusätzlich ein Glucagon-Like Peptide-1 (GLP1 ) oder ein Analogon oder Derivat davon, oder Exendin-3 bzw. -4 oder ein Analogon oder Derivat davon enthalten ist. A further subject of the invention is a pharmaceutical formulation as described above, which additionally contains a glucagon-like peptide-1 (GLP1) or an analogue or derivative thereof, or exendin-3 or -4 or an analogue or derivative thereof ,
Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, bei dem zusätzlich Exendin-4 enthalten ist. Another object of the invention is a pharmaceutical formulation as described above, which additionally contains exendin-4.
Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, bei dem ein Analogon von Exendin-4 ausgewählt wird aus einer Gruppe enthaltend Another object of the invention is a pharmaceutical formulation as described above in which an analog of exendin-4 is selected from a group containing
H-desPro36-Exendin-4-Lys6-NH2, H-des(Pro3637)-Exendin-4-Lys4-NH2 und H-desPro 36 -Exendin-4-Lys 6 -NH 2 , H-des (Pro 3637 ) -Exendin-4-Lys 4 -NH 2 and
H-des(Pro3637)-Exendin-4-Lys5-NH2, H des (Pro 3637 ) -Exendin-4-Lys 5 -NH 2 ,
oder ein pharmakologisch tolerierbares Salz davon. Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, bei dem ein Analogon von Exendin-4 ausgewählt wird aus einer Gruppe enthaltend or a pharmacologically tolerable salt thereof. Another object of the invention is a pharmaceutical formulation as described above in which an analog of exendin-4 is selected from a group containing
desPro36 [Asp28jExendin-4 (1-39), desPro 36 [Asp 28 jExendin-4 (1-39),
desPro36 [lsoAsp2δjExendin-4 (1-39), desPro 36 [lsoAsp 2δ jExendin-4 (1-39),
desPro36 [Met(O)14, Asp28jExendin-4 (1-39), desPro 36 [Met (O) 14 , Asp 28, jExendin-4 (1-39),
desPro36 [Met(O)14, lsoAsp28]Exendin-4 (1-39), desPro 36 [Met (O) 14 , IsoPs 28 ] Exendin-4 (1-39),
desPro36 [Trp(O2)25, Asp2δ]Exendin-2 (1-39), desPro36 [Trp(O2)25, lsoAsp28]Exendin-2 (1-39), desPro 36 [Trp (O 2 ) 25 , Asp 2δ ] Exendin -2 (1-39), the Pro 36 [Trp (O 2 ) 25 , IsoPs 28 ] Exendin-2 (1-39),
desPro36 [Met(O)14Trp(O2)25, Asp28]Exendin-4 (1-39) und desPro 36 [Met (O) 14 Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1-39) and
desPro36 [Met(O)14Trp(O2)25, lsoAsp28]Exendin-4 (1-39), desPro 36 [Met (O) 14 Trp (O 2 ) 25 , IsoPs 28 ] Exendin-4 (1-39),
oder ein pharmakologisch tolerierbares Salz davon. or a pharmacologically tolerable salt thereof.
Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, bei denen an die C-Termini der Analoga von Exendin-4 das Peptid -LySo-NH2 angefügt ist. Another object of the invention is a pharmaceutical formulation as described above, in which the peptide -LySo-NH 2 is added to the C-termini of the analogues of exendin-4.
Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, bei dem ein Analogon von Exendin-4 ausgewählt wird aus einer Gruppe enthaltend Another object of the invention is a pharmaceutical formulation as described above in which an analog of exendin-4 is selected from a group containing
H-(LyS)6- des Pro36 [Asp28]Exendin-4(1-39)-Lys6-NH2 H- (LyS) 6 - of Pro 36 [Asp 28 ] Exendin-4 (1-39) -Ly 6 -NH 2
des Asp28Pro36, Pro37, Pro38 Exendin-4(1-39) -NH2, Asp 28 Pro 36 , Pro 37 , Pro 38 Exendin-4 (1-39) -NH 2 ,
H-(LyS)6- des Pro36, Pro37, Pro38 [Asp28]Exendin-4(1-39) -NH2, H- (LyS) 6 - of Pro 36 , Pro 37 , Pro 38 [Asp 28 ] Exendin-4 (1-39) -NH 2 ,
H-Asn-(Glu)5 des Pro36, Pro37, Pro38 [Asp28]Exendin-4(1-39) -NH2, H-Asn- (Glu) 5 of Pro 36 , Pro 37 , Pro 38 [Asp 28 ] Exendin-4 (1-39) -NH 2 ,
des Pro36, Pro37, Pro38 [Asρ28]Exendin-4(1-39)-(Lys)6-NH2, Pro 36 , Pro 37 , Pro 38 [Asρ 28 ] Exendin-4 (1-39) - (Lys) 6 -NH 2 ,
H-(LyS)6- des Pro36, Pro37, Pro38 [Asp28jExendin-4(1-39)-(Lys)6-NH2, H- (LyS) 6 - of Pro 36 , Pro 37 , Pro 38 [Asp 28 jExendin-4 (1-39) - (Lys) 6 -NH 2 ,
H-Asn-(Glu)5- des Pro36, Pro37, Pro38 [Asp28]Exendin-4(1-39)-(Lys)6-NH2, H-(LyS)6- des Pro36 [Trp(O2)25, Asp28]Exendin-4(1-39)-Lys6-NH2, H-Asn- (Glu) 5 - of Pro 36 , Pro 37 , Pro 38 [Asp 28 ] Exendin-4 (1-39) - (Lys) 6 -NH 2 , H- (LyS) 6 - of Pro 36 [Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1-39) -Lys 6 -NH 2 ,
H- des Asp28 Pro36, Pro37, Pro38 [Trp(O2)25]Exendin-4(1-39) -NH2, H- the Asp 28 Pro 36, Pro 37, Pro 38 [Trp (O2) 25] Exendin-4 (1-39) -NH 2,
H-(LyS)6- des Pro36, Pro37, Pro38 [Trp(O2)25, Asp28]Exendin-4(1-39) -NH2, H- (LyS) 6 - of Pro 36 , Pro 37 , Pro 38 [Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1-39) -NH 2 ,
H-ASn-(GIu)5- des Pro36, Pro37, Pro38 [Trp(O2)25, Asp28]Exendin-4(1 -39) -NH2, des Pro36, Pro37, Pro38 [Trp(O2)25, Asp28]Exendin-4(1-39)-(Lys)6-NH2, H-ASn (Glu) 5 - of Pro 36 , Pro 37 , Pro 38 [Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1 -39) -NH 2 , Pro 36 , Pro 37 , Pro 38 [Trp (O 2 ) 25 , Asp 28 ] exendin-4 (1-39) - (Lys) 6 -NH 2 ,
H-(LyS)6- des Pro36, Pro37, Pro38 [Trp(O2)25, Asp28]Exendin-4(1-39)-(Lys)6-NH2, H-Asn-(Glu)5- des Pro36, Pro37, Pro38 [Trp(O2)25, Asp28]Exendin-4(1-39)-(Lys)6-NH2, H-(LyS)6- des Pro36 [Met(O)14, Asρ28]Exendin-4(1 -39)-Lys6-NH2, H- (LyS) 6 - of Pro 36 , Pro 37 , Pro 38 [Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1-39) - (Lys) 6 -NH 2 , H-Asn- (Glu ) 5 - of Pro 36 , Pro 37 , Pro 38 [Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1-39) - (Lys) 6 -NH 2 , H- (LyS) 6 - of Pro 36 [Met (O) 14 , Asρ 28 ] exendin-4 (1 -39) -Lys 6 -NH 2 ,
des Met(O)14 Asp28 Pro 36, Pro37, Pro38 Exendin-4(1-39) -NH2, the Met (O) 14 Asp 28 Pro 36, Pro 37, Pro 38 exendin-4 (1-39) -NH 2,
H-(LyS)6- des Pro36, Pro 37, Pro38 [Met(O)14, Asp28]Exendin-4(1-39) -NH2, H- (LyS) 6 - of Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Asp 28 ] Exendin-4 (1-39) -NH 2 ,
H-ASn-(GIu)5- des Pro36, Pro37, Pro38 [Met(O)14, Asp28] Exendin-4(1 -39) -NH2, des Pro36, Pro37, Pro38 [Met(O)14, Asp28]Exendin-4(1-39)-(Lys)6-NH2, H-ASn (Glu) 5 - of Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Asp 28 ] Exendin-4 (1 -39) -NH 2 , of Pro 36 , Pro 37 , Pro 38 [ Met (O) 14 , Asp 28 ] Exendin-4 (1-39) - (Lys) 6 -NH 2 ,
H-(LyS)6- des Pro36, Pro37, Pro38 [Met(O)14, Asp28jExendin-4(1-39)-Lys6-NH2, H- (LyS) 6 - of Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Asp 28, jExendin-4 (1-39) -Lys 6 -NH 2 ,
H-Asn-(Glu)5 des Pro36, Pro37, Pro38 [Met(O)14, Asp28] Exendin-4(1-39)-(Lys)6-NH2, H-Asn- (Glu) 5 of Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Asp 28 ] Exendin-4 (1-39) - (Lys) 6 -NH 2 ,
H-(LyS)6- des Pro36 [Met(O)14, Trp(O2)25, Asp28jExendin-4(1-39)-Lys6-NH2, H- (LyS) 6 - of Pro 36 [Met (O) 14 , Trp (O 2 ) 25 , Asp 28 ] exendin-4 (1-39) -Lys 6 -NH 2 ,
des Asp28 Pro36, Pro37, Pro38 [Met(O)14, Trp(O2)25]Exendin-4(1-39) -NH2, Asp 28 Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Trp (O 2 ) 25 ] Exendin-4 (1-39) -NH 2 ,
H-(LyS)6- des Pro36' Pro37, Pro38 [Met(O)14, Trp(O2)25, Asp28]Exendin-4(1-39) -NH2, H-Asn-(Glu)5- des Pro36, Pro37, Pro38 [Met(O)14, Asp28] Exendin-4(1-39) -NH2, des Pro36, Pro37, Pro38 [Met(O)14, Trp(O2)25, Asp28]Exendin-4(1-39)-(Lys)6-NH2, H-(LyS)6- des Pro36' Pro37, Pro38 [Met(O)14, Trp(O2)25, Asp28]Exendin-4(1 -39)-(Lys)6- NH2, H- (LyS) 6 - of Pro 36 'Pro 37 , Pro 38 [Met (O) 14 , Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1-39) -NH 2 , H-Asn- ( Glu) 5 - Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Asp 28 ] Exendin-4 (1-39) -NH 2 , Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1-39) - (Lys) 6 -NH 2 , H- (LyS) 6 - of Pro 36 'Pro 37 , Pro 38 [Met (O) 14 , Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1 -39) - (Lys) 6 - NH 2 ,
H-ASn-(GIu)5- des Pro36, Pro37, Pro38 [Met(O)14, Trp(O2)25, Asp28] Exendin-4(1-39)- (LyS)6-NH2, H-ASn- (Glu) 5 - of Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1-39) - (LyS) 6 -NH 2 ,
oder ein pharmakologisch tolerierbares Salz davon. or a pharmacologically tolerable salt thereof.
Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, bei dem zusätzlich Arg34, Lys26 (Nε(γ-glutamyl(Nα-hexadecanoyl))) GLP-1 (7-37) [liraglutide] oder ein pharmakologisch tolerierbares Salz davon enthalten ist. Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, bei der die Aminosäure Methionin vorhanden ist, vorzugsweise im Konzentrationsbereich von bis zu 10 mg/mi, besonders bevorzugt von bis zu 3 mg/ml. Ein weiterer Gegenstand der Erfindung ist ein Verfahren zur Herstellung einer Another object of the invention is a pharmaceutical formulation as described above, in which in addition Arg 34 , Lys 26 (N ε (γ-glutamyl (N α -hexadecanoyl))) GLP-1 (7-37) [liraglutide] or a pharmacologically tolerable salt thereof is included. Another object of the invention is a pharmaceutical formulation as described above, in which the amino acid methionine is present, preferably in the concentration range of up to 10 mg / ml, more preferably of up to 3 mg / ml. Another object of the invention is a method for producing a
Formulierung wie oben beschrieben, bei dem Formulation as described above, in which
(a) die Komponenten in eine wässrige Lösung eingebracht werden und  (a) the components are introduced into an aqueous solution and
(b) der pH-Wert eingestellt wird. Ein weiterer Gegenstand der Erfindung ist eine Verwendung einer Formulierung wie oben beschrieben zur Behandlung von Diabetes Mellitus.  (b) the pH is adjusted. Another object of the invention is a use of a formulation as described above for the treatment of diabetes mellitus.
Ein weiterer Gegenstand der Erfindung ist ein Arzneimittel zur Behandlung von Another object of the invention is a medicament for the treatment of
Diabetes Mellitus bestehend aus einer Formulierung wie oben beschrieben. Diabetes mellitus consisting of a formulation as described above.
Die Zubereitung kann desweiteren Konservierungsmittel (z.B. Phenol, Kresol, The preparation may further contain preservatives (e.g., phenol, cresol,
Parabene), Isotonisierungsmittel (z.B. Mannitol, Sorbitoi, Lactose, Dextrose, Parabens), isotonizing agents (e.g., mannitol, sorbitoi, lactose, dextrose,
Trehalose, Natriumchlorid, Glycerol). Puffersubstanzen, Salze, Säuren und Laugen sowie weitere Hilfstoffe enthalten. Diese Substanzen können jeweils einzeln oder auch als Mischungen vorliegen. Trehalose, sodium chloride, glycerol). Buffer substances, salts, acids and alkalis and other excipients. These substances may be present individually or as mixtures.
Giycerol, Dextrose, Lactose, Sorbitoi und Mannitol liegen üblicherweise in der pharmazeutischen Zubereitung in einer Konzentration von 100 - 250 mM, NaCI in einer Konzentration bis zu 150 mM vor. Puffersubstanzen, wie z.B. Phosphat-, Acetat-, Citrat, Arginin , Glycylglycin oder TRIS (d.h. 2-Amino-2-Hydroxymethyi-1 ,3,-Giycerol, dextrose, lactose, sorbitoi and mannitol are usually present in the pharmaceutical preparation in a concentration of 100-250 mM, NaCl in a concentration up to 150 mM. Buffer substances, e.g. Phosphate, acetate, citrate, arginine, glycylglycine or TRIS (i.e., 2-amino-2-hydroxymethyl-1, 3, -
Propandiol) Puffer sowie entsprechende Salze, können in einer Konzentration von 5 - 250 mM, bevorzugt 10 - 100 mM vorhanden sein. Weitere Hilfstoffe können unter anderem sein Salze oder Arginin. Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben beschrieben, bei der das Insulinanalogon in einer Konzentration von 60 - 6000 nmol/ml (dies entspricht etwa einer Konzentration von 0,35 - 70 mg/ml oder 10 - 1000 Einheiten/ml), vorzugsweise in einer Konzentration von 240 - 3000 nmol/mi vorliegt (dies entspricht etwa einer Konzentration von 1 ,4 - 35 mg/ml oder 40 - 500 Propandiol) buffers and corresponding salts may be present in a concentration of 5-250 mM, preferably 10-100 mM. Other excipients may include salts or arginine. A further subject of the invention is a pharmaceutical formulation as described above in which the insulin analogue is in a concentration of 60-6000 nmol / ml (this corresponds approximately to a concentration of 0.35-70 mg / ml or 10-1000 Units / ml), preferably in a concentration of 240-3000 nmol / ml (this corresponds approximately to a concentration of 1.4-35 mg / ml or 40-500
Einheiten/ml); Units / ml);
bei der das Tensid in einer Konzentration von 5 - 200 μg/ml, bevorzugt von 5 - 120μg/ml und besonders bevorzugt von 20 - 75 μg/ml vorliegt. in which the surfactant is present in a concentration of 5-200 μg / ml, preferably of 5-120 μg / ml and more preferably of 20-75 μg / ml.
Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben ausgeführt, bei der Glycerol und/oder Mannitol in einer Konzentration von 100 - 250 mM, und/oder NaCI vorzugsweise in einer Konzentration bis zu 150 mM vorliegt. Another object of the invention is a pharmaceutical formulation as stated above, in which glycerol and / or mannitol in a concentration of 100-250 mM, and / or NaCl preferably in a concentration up to 150 mM.
Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Formulierung wie oben ausgeführt, bei der eine Puffersubstanz in einer Konzentration von 5 - 250 mM vorliegt. Ein weiterer Gegenstand der Erfindung ist eine pharmazeutische Insulinformulierung, die weitere Zusätze wie z.B. Salze enthält, welche die Freigabe von Insulin retardieren. Auch Mischungen aus solchen Retardinsulinen mit oben beschriebenen Another object of the invention is a pharmaceutical formulation as stated above, in which a buffer substance in a concentration of 5 - 250 mM is present. Another object of the invention is a pharmaceutical insulin formulation, the further additives such. Contains salts which retard the release of insulin. Also mixtures of such sustained-release insulins described above
Formulierungen sind darin eingeschlossen. Ein weiterer Gegenstand der Erfindung ist eine Methode zur Herstellung solcher pharmazeutischer Formulierungen. Ebenso ist ein weiterer Gegenstand der Erfindung die Verwendung solcher Formulierungen zur Behandlung von Diabetes mellitus. Formulations are included. Another object of the invention is a method for producing such pharmaceutical formulations. Likewise, another object of the invention is the use of such formulations for the treatment of diabetes mellitus.
Ein weiterer Gegenstand der Erfindung ist die Verwendung bzw. der Zusatz von Tensiden als Stabilisator während des Herstellungsprozesses von Insulin, Another object of the invention is the use or the addition of surfactants as a stabilizer during the production process of insulin,
Insulinanaloga oder Insulinderivaten oder deren Zubereitungen. Insulin analogues or insulin derivatives or their preparations.
Im folgenden wird die Anmeldung anhand einiger Beispiele beschrieben, die  In the following the application will be described by means of some examples which
keinesfalls beschränkend wirken sollen. by no means restrictive.
Beispiele: Die folgenden Beispiele sollen den Erfindungsgedanken näher erläutern, ohne beschränkend zu wirken. Examples: The following examples are intended to explain the concept of the invention in more detail without being restrictive.
Figurenlegende: Figure legend:
Fig. 1 : Blutzuckersenkende Wirkung von neuen Insulinanaloga gemäß Formel I inFig. 1: Hypoglycemic effect of new insulin analogues according to formula I in
Ratten rats
Fig. 2: Blutzuckersenkende Wirkung von neuen Insulinanaloga gemäß Formel I imFig. 2: blood sugar lowering effect of new insulin analogues according to formula I in
Hund dog
Fig. 3: Blutzuckersenkende Wirkung von YKL205 im Hund  Fig. 3: Hypoglycemic effect of YKL205 in the dog
Fig. 4: Zinkabhängigkeit der hypoglykämischen Wirkung von YKL205 im Hund Fig. 4: Zinc dependence of the hypoglycemic effect of YKL205 in the dog
Beispiel 1 : Studien zur Evaluierung des optimalen pH Werts Example 1: Studies to evaluate the optimal pH value
Die Herstellung der Lösung erfolgt indem ca. 25 % Wasser für Injektionszwecke vorgelegt werden. Nacheinander werden SAR161271 und die Zinkchlorid The solution is prepared by adding about 25% water for injection. One after the other are SAR161271 and the zinc chloride
Stammlösung hinzugefügt und gerührt. Durch Zugabe von 1 M HCl bei einem pH Wert von pH 2 wird SAR161271 gelöst. Die Lösung wird gerührt, dann erfolgt die Zugabe von 1 M NaOH zur Einstellung des pH Werts auf pH 3.75 (3.8). Mit Wasser für  Stock solution added and stirred. By adding 1 M HCl at a pH of pH 2, SAR161271 is dissolved. The solution is stirred, then the addition of 1 M NaOH to adjust the pH to pH 3.75 (3.8). With water for
Injektionszwecke wird auf 90 % der Ansatzgröße aufgefüllt. Zu der Lösung Injections are made up to 90% of the batch size. To the solution
nacheinander Glycerol 85 % und m-Cresol unter Rühren dazugeben. Mit Wasser für Injektionszwecke auf das gewünschte Endgewicht auffüllen. Lösung mittels successively add glycerol 85% and m-cresol with stirring. Make up to the desired final weight with water for injections. Solution by means of
Spritzenvorsatzfilter filtrieren. Mittels dieser Art der Lösungsherstellung wurden Formulierungen hergestellt, die auf folgende pH Werte eingestellt wurden: pH 3.0, 3.25, 3.5, 3.75, 4.0 und 4.5. Mit diesen Formulierungen wurde eine 3 Monats Filter syringe filter. By means of this type of solution preparation, formulations were prepared which were adjusted to the following pH values: pH 3.0, 3.25, 3.5, 3.75, 4.0 and 4.5. With these formulations was a 3 month
Stabilitäts-Studie durchgeführt. Im Folgenden werden die 2 Monats Stabilitäts-Studien Ergebnisse der Formulierungen von pH 3.0, 4.0 und 4.5 aufgeführt. t o τ2 Monate; 5°C t 2 Monate: 25°C 60% rH pH 3.5 pH 4.0 pH 4.5 pH 3.5 pH 4.0 pH 4.5 pH 3.5 pH 4.0 pH 4.5 pH 3.5 4.0 4.5 3.5 4.0 4.5 3.5 4.0 4.5Stability study conducted. In the following, the 2 month stability studies results of the formulations of pH 3.0, 4.0 and 4.5 are listed. to τ2 months; 5 ° C t 2 months: 25 ° C 60% RH pH 3.5 pH 4.0 pH 4.5 pH 3.5 pH 4.0 pH 4.5 pH 3.5 pH 4.0 pH 4.5 pH 3.5 4.0 4.5 3.5 4.0 4.5 3.5 4.0 4.5
Λssay 3.87 3.81 3.68 3.71 3.80 3.69 3.76 3.72 3.53Λssay 3.87 3.81 3.68 3.71 3.80 3.69 3.76 3.72 3.53
SΛR 171271 SΛR 171271
[mg/ml]  [Mg / ml]
Andere älinlichc 0.6 / 2.8 0.6 / 2.8 0.6 / 2.9 0.7 / 2.7 0.7 / 2.9 1.0 / 3.4 1.0 / 3.1 1.1 / 4. 1 1.8 / Other älinlichc 0.6 / 2.8 0.6 / 2.8 0.6 / 2.9 0.7 / 2.7 0.7 / 2.9 1.0 / 3.4 1.0 / 3.1 1.1 / 4. 1 1.8 /
Verunreinigungc 7.2 n / älinlichc Contamination 7.2
Veranreiniguiigc  Veranreiniguiigc
ii gesamt [%] ii total [%]
Assay m-Crcsol 2.8 2.5 2.9 2.7 2.5 2.8 2.7 2.4 2.8 Assay m-Crcsol 2.8 2.5 2.9 2.7 2.5 2.8 2.7 2.4 2.8
[mg/ml] [Mg / ml]
HMWP [%] 0.2 0.2 0.2 0.3 0.3 0.3 0.3 0.4 1.0 HMWP [%] 0.2 0.2 0.2 0.3 0.3 0.3 0.3 0.4 1.0
Trübung klar klar klar klar klar klar klar klar klar Cloudy clear clear clear clear clear clear clearly clear
Die Ergebnisse zeigen, dass je aeider der pH Wert der Lösung ist, umso stabiler ist diese. The results show that the more the pH of the solution is, the more stable it is.
Beispiel 2: Studien zur Wahl des Tonizitäts Agens Example 2: Studies on the choice of the tonicity agent
Die Herstellung der Lösung wurde, wie in Beispiel 1 beschrieben, durchgeführt. Die Ergebnisse der 2 Monats Stabilitäts-Studie 2.5 % Glycerol vs. 0.8 % NaCI sind wie folgt. The preparation of the solution was carried out as described in Example 1. The results of the 2 month stability study 2.5% glycerol vs. 0.8% NaCl is as follows.
2.5 % Glycerol 2.5% glycerol
Gehalt SAR161271  Salary SAR161271
2 M + 5° C: 3.56 mg/ml  2 M + 5 ° C: 3.56 mg / ml
2 M + 25° C: 3.46 mg/ml  2 M + 25 ° C: 3.46 mg / ml
Verunreinigungen  impurities
2 M + 5° C: 2.6 %  2 M + 5 ° C: 2.6%
2 M + 25° C: 3.8 %  2 M + 25 ° C: 3.8%
Höher molekulargewichtige Proteine  Higher molecular weight proteins
2 M + 5° C: 0.2 %  2 M + 5 ° C: 0.2%
2 M + 25° C: 0.4 %  2 M + 25 ° C: 0.4%
0.8 % NaCI 0.8% NaCl
Gehalt SAR161271  Salary SAR161271
2 M + 5° C: 3.52 mg/ml 2 M + 25° C: 3.49 mg/ml 2 M + 5 ° C: 3.52 mg / ml 2 M + 25 ° C: 3.49 mg / ml
Verunreinigungen  impurities
2 M + 5° C: 2.7 %  2 M + 5 ° C: 2.7%
2 M + 25° C: 4.8 %  2 M + 25 ° C: 4.8%
Höher molekulargewichtige Protein Higher molecular weight protein
2 M + 5° C: 0.2 %  2 M + 5 ° C: 0.2%
2 M + 25° C: 1.1 %  2 M + 25 ° C: 1.1%
Mit Hiife der Ergebnisse dieser Studie wurde das Tonizitätsagens Glyceroi in der Konzentration 2.5 % ausgewählt. Die Formulierung ist stabiler verglichen mit 0.8 % NaCI als Tonizitätsagens. Zusätzlich wurde eine Präzipitation bei Verwendung von NaCI während der Herstellung festgestellt, die unlöslich war. Die Osmolarität betrug bei beiden Substanzen 290 ± 30 mθsmol/kg.  With the help of the results of this study, the tonicity agent Glyceroi in the concentration 2.5% was selected. The formulation is more stable compared to 0.8% NaCl as a tonicity agent. In addition, precipitation was detected using NaCl during the preparation which was insoluble. The osmolarity was 290 ± 30 mθsmol / kg for both substances.
Beispiel 3: Studien zur Auswahl des Konservierungsmittels Example 3: Studies on Preservative Selection
Die im Folgenden beschriebenen Formulierungen wurden, wie in Beispiel 1 The formulations described below were as in Example 1
Beschrieben, hergestellt. Formulierungen mit verschiedenen Konservierungsmitteln wurden zur mikrobiellen Qualitätskontrolle gegeben und dort einem Described, made. Formulations with different preservatives were given for microbial quality control and there one
Konservierungsmittelbelastungstest unterzogen (entsprechend Ph. Eur. 5.5 Kriterium A und USP 29).  Subject to preservative loading test (according to Ph. Eur. 5.5 Criterion A and USP 29).
m-Cresol: 1.5, 1.8, 2.1 und 2.7 mg/ml m-cresol: 1.5, 1.8, 2.1 and 2.7 mg / ml
Phenol: 2.7 mg/ml Phenol: 2.7 mg / ml
m-Cresol: 1.5 mg/ml and Phenol: 0.6 mg/ml I m-cresol: 1.5 mg / ml and phenol: 0.6 mg / ml I
Benzyl alcohol: 15 mg/ml Benzyl alcohol: 15 mg / ml
m-Cresol wurde als Konservierungsmittel ausgewählt. Die Konzentration von 2 mg/ml wurde gewählt, obwohl bereits 1.5 mg/ml zur Konservierung ausreichend gewesen wären. Dennoch wurde die höhere m-Cresol Konzentration gewählt aufgrund mikrobiologischer Sicherheitsaspekte und der Spezifikationfestlegung. Zusätzlich wurden die Formulierungen (außer 2.1 mg/ml m-Cresol) auf Stabilität (3 Monate) gelegt. Beispiel 4: Formulierung der amidierten Insulinderivate m-cresol was chosen as a preservative. The concentration of 2 mg / ml was chosen, although 1.5 mg / ml would have been sufficient for preservation. Nevertheless, the higher m-cresol concentration was chosen due to microbiological safety aspects and specification specification. In addition, the formulations (except 2.1 mg / ml m-cresol) were placed on stability (3 months). Example 4: Formulation of the amidated insulin derivatives
Die Beispiele 4 bis 8 dienen nur zur Bestimmung der biologischen, pharmakologischen und physikalisch-chemischen Eigenschaften von Insulinanaloga gemäß Formel I indem zunächst Formulierungen davon bereitgestellt wurden (Beispiel 4) und dann entsprechende Tests vorgenommen wurden (Beispiele 5 bis 8). Es wurde von den Verbindungen wie folgt eine Lösung hergestellt: Das erfindungsgemäße Insulinanalog wurde mit einer Zielkonzentration von 240 ± 5 μM in 1 mM Salzsäure mit 80 μg/mL Zink (als Zinkchlorid) aufgelöst. Examples 4 to 8 serve only to determine the biological, pharmacological and physicochemical properties of insulin analogues according to formula I by firstly providing formulations thereof (example 4) and then carrying out corresponding tests (examples 5 to 8). A solution was prepared of the compounds as follows: The insulin analog according to the invention was dissolved at a target concentration of 240 ± 5 μM in 1 mM hydrochloric acid with 80 μg / ml zinc (as zinc chloride).
Als Lösungsmedium wurden folgende Zusammensetzungen verwendet: The following compositions were used as solvent medium:
a) 1 mM Salzsäure a) 1 mM hydrochloric acid
b) 1 mM Salzsäure, 5 μg/mL Zink (als Zinkchlorid oder Salzsäure zugesetzt) c) 1 mM Salzsäure, 10 μg/mL Zink (als Zinkchlorid oder Salzsäure zugesetzt) d) 1 mM Salzsäure, 15 μg/mL Zink (als Zinkchlorid oder Salzsäure zugesetzt) e) 1 mM Salzsäure, 30 μg/mL Zink (als Zinkchlorid oder Salzsäure zugesetzt) f) 1 mM Salzsäure, 80 μg/mL Zink (als Zinkchlorid oder Salzsäure zugesetzt) g) 1 mM Salzsäure, 120 μg/mL Zink (als Zinkchlorid oder Salzsäure zugesetzt) b) 1 mM hydrochloric acid, 5 μg / ml zinc (added as zinc chloride or hydrochloric acid) c) 1 mM hydrochloric acid, 10 μg / ml zinc (added as zinc chloride or hydrochloric acid) d) 1 mM hydrochloric acid, 15 μg / ml zinc (as zinc chloride or hydrochloric acid added) e) 1 mM hydrochloric acid, 30 μg / ml zinc (added as zinc chloride or hydrochloric acid) f) 1 mM hydrochloric acid, 80 μg / ml zinc (added as zinc chloride or hydrochloric acid) g) 1 mM hydrochloric acid, 120 μg / ml Zinc (added as zinc chloride or hydrochloric acid)
Hierzu wurde von dem gefriergetrockneten Material zunächst eine um etwa 30% höhere Menge als aufgrund des Molekulargewichts und der angestrebten For this purpose, of the freeze-dried material was first about a 30% higher amount than due to the molecular weight and the desired
Konzentration benötigt eingewogen. Danach wurde die vorliegende Konzentration mittels analytischer HPLC bestimmt und die Lösung anschließend auf das zur Concentration needed weighed. Thereafter, the present concentration was determined by analytical HPLC and the solution was then added to the
Erreichung der Zielkonzentration erforderliche Volumen mit 5 mM Salzsäure mit 80 μg/mL Zink aufgefüllt. Falls erforderlich, wurde dabei der pH-Wert auf 3,5 ± 0,1 nachjustiert. Nach der endgültigen Analyse durch HPLC zur Absicherung der Achievement of the target concentration volume required with 5 mM hydrochloric acid replenished with 80 μg / mL zinc. If necessary, the pH was readjusted to 3.5 ± 0.1. After the final analysis by HPLC to secure the
Zielkonzentration von 240 ± 5 μM wurde die fertige Lösung mittels einer Spritze mit einem 0,2 μm Filtervorsatz in ein mit einem Septum und einer Bördelkappe Target concentration of 240 ± 5 μM, the final solution was syringed with a 0.2 μm filter attachment into a with a septum and a crimp cap
verschlossenes steriles Fläschchen überführt. Für die kurzfristige, einmalige Testung der erfindungsgemäßen Insulinderivate wurde keine Optimierung der Formulierungen, z.B. hinsichtlich eines Zusatzes isotonischer Agentien, Konservierungsmittel oder Puffersubstanzen, vorgenommen. transferred sterile sterile vial. For the short-term, one-time testing of the insulin derivatives according to the invention no optimization of the formulations, eg with regard to an addition of isotonic agents, preservatives or buffer substances.
Beispiel 5: Evaluierung der blutzuckersenkenden Wirkung von neuen Insulinanaloga in der Ratte Example 5: Evaluation of the hypoglycemic effect of new insulin analogues in the rat
Die blutzuckersenkende Wirkung von ausgewählten neuen Insulinanaloga wird in männlichen, gesunden, normoglykämischen Wistarratten geprüft. Männlichen Ratten wird eine Dosis von 9 nmol/kg eines Insulinanalogons subkutan injiziert. Unmittelbar vor der Injektion des Insulinanalogons und in regelmäßigen Abständen bis zu acht Stunden nach der Injektion werden den Tieren Blutproben entnommen und darin der Blutzuckergehalt bestimmt. Das Experiment zeigt deutlich (vgl. Fig. 1 ), dass das eingesetzte erfindungsgemäße Insulinanalogon zu einem deutlich verzögerten The hypoglycemic effect of selected new insulin analogues is tested in male, healthy, normoglycemic Wistar rats. Male rats are injected subcutaneously with a dose of 9 nmol / kg of an insulin analogue. Immediately before injecting the insulin analogue and periodically up to eight hours after the injection, blood samples are taken from the animals and the blood sugar content is determined therein. The experiment clearly shows (see Fig. 1) that the insulin analog used according to the invention significantly delayed
Wirkeintritt und einer längeren, gleichmäßigen Wirkdauer führt. Effective and a longer, uniform duration of action leads.
Beispiel 6: Evaluierung der blutzuckersenkenden Wirkung von neuen Insulinanaloga im Hund  Example 6: Evaluation of the hypoglycaemic effect of new insulin analogues in the dog
Die blutzuckersenkende Wirkung von ausgewählten neuen Insulinanaloga wird in männlichen, gesunden, normoglykämischen Beaglehunden geprüft. Männlichen Tieren wird eine Dosis von 6 nmol/kg eines Insulinanalogons subkutan injiziert. Unmittelbar vor der Injektion des Insulinanalogons und in regelmäßigen Abständen bis zu achtundvierzig Stunden nach der Injektion werden den Tieren Blutproben entnommen und darin der Blutzuckergehalt bestimmt. Das Experiment zeigt deutlich (vgl. Fig. 2), dass das eingesetzte erfindungsgemäße Insulinanalogon zu einem deutlich The hypoglycemic effect of selected new insulin analogues is tested in male, healthy, normoglycemic beagle dogs. Male animals are injected subcutaneously with a dose of 6 nmol / kg of an insulin analogue. Immediately before injecting the insulin analogue and at regular intervals up to forty-eight hours after the injection, blood samples are taken from the animals and the blood sugar content is determined therein. The experiment clearly shows (see Fig. 2) that the insulin analog according to the invention used is a clear one
verzögerten Wirkeintritt und einer längeren, gleichmäßigen Wirkdauer führt. delayed onset of action and a longer, uniform duration of action leads.
Beispiel 7: Evaluierung der blutzuckersenkenden Wirkung am Hund bei Example 7: Evaluation of the hypoglycemic effect in dogs
zweifach erhöhter Dosis Die blutzuckersenkende Wirkung von ausgewählten neuen Insulinanaloga wird in männlichen, gesunden, normoglykämischen Beaglehunden geprüft. Männlichen Tieren wird eine Dosis von 6 nmol/kg und 12nmol/kg eines Insulinanalogons subkutan injiziert. Unmittelbar vor der Injektion des Insulinanalogons und in regelmäßigen Abständen bis zu achtundvierzig Stunden nach der Injektion werden den Tieren Blutproben entnommen und darin der Blutzuckergehalt bestimmt. twice the dose The hypoglycemic effect of selected new insulin analogues is tested in male, healthy, normoglycemic beagle dogs. Male animals are injected subcutaneously with a dose of 6 nmol / kg and 12 nmol / kg of an insulin analogue. Immediately before injecting the insulin analogue and at regular intervals up to forty-eight hours after the injection, blood samples are taken from the animals and the blood sugar content is determined therein.
Das Experiment zeigt deutlich (vgl. Fig. 3), dass das eingesetzte erfindungsgemäße Insulinanalogon dosisabhängig wirkt, dass aber trotz zweifach erhöhter Dosis der Wirkungsverlauf flach verläuft, d.h. kein ausgeprägter Tiefpunkt (Nadir) beobachtet wird. Daraus lässt sich ableiten, dass die erfindungsgemäßen Insuline im Vergleich zu bekannten Verzögerungsinsulinen zu deutlich weniger hypoglykämischen Ereignissen führen. The experiment clearly shows (see Fig. 3) that the insulin analogue according to the invention used has a dose-dependent effect, but that despite a doubly increased dose, the course of action is flat, ie. no pronounced nadir is observed. It can be deduced from this that the insulins according to the invention lead to significantly fewer hypoglycemic events in comparison with known delay insulins.
Beispiel 8: Evaluierung der blutzuckersenkenden Wirkung am Hund bei verschiedenen Zinkkonzentrationen in der Formulierung Example 8: Evaluation of the hypoglycemic effect in dogs at different zinc concentrations in the formulation
Die Experimente wurden, wie in Beispiel 35 beschrieben, durchgeführt. Figur 4 zeigt das Ergebnis. Danach lässt sich die Zeit - Wirkungskurve des erfindungsgemäßen Insulinanalogons durch den Gehalt an Zinkionen in der Formulierung bei gleicher Insulinkonzentration in der Weise beeinflussen, dass man bei null oder geringem Zinkgehalt einen schnellen Wirkungseintritt beobachtet und die Wirkung über 24 Stunden anhält, während bei höherem Zinkgehalt ein flacher Wirkungseintritt beobachtet wird und die Insulinwirkung deutlich länger als 24 Stunden anhält. The experiments were carried out as described in Example 35. FIG. 4 shows the result. Thereafter, the time-effect curve of the insulin analog according to the invention by the content of zinc ions in the formulation at the same concentration of insulin influence in such a way that at zero or low zinc content observed a rapid onset and the effect persists over 24 hours, while at higher zinc content low onset of action is observed and the insulin action persists well longer than 24 hours.
Beispiel 9: Formulierung der amidierten Insulinderivate Example 9: Formulation of the amidated insulin derivatives
Die Beispiele 9 bis 11 dienen nur zur Bestimmung der biologischen, Examples 9 to 11 are only for the determination of biological,
pharmakologischen und physikalisch-chemischen Eigenschaften von Insulinanaloga gemäß Formel Il indem zunächst Formulierungen davon bereitgestellt wurden pharmacological and physicochemical properties of insulin analogues according to formula II by first providing formulations thereof
(Beispiel 9) und dann entsprechende Tests vorgenommen wurden (Beispiele 10 und 11 ). Das erfindungsgemäße Insulinanalog wurde mit einer Zielkonzentration von 240 + 5 μM in 1 mM Salzsäure mit 80 μg/mL Zink (als Zinkchlorid) aufgelöst. Hierzu wurde von dem gefriergetrockneten Material zunächst eine um etwa 30% höhere Menge als aufgrund des Molekulargewichts und der angestrebten Konzentration benötigt eingewogen. Danach wurde die vorliegende Konzentration mittels analytischer HPLC bestimmt und die Lösung anschließend auf das zur Erreichung der Zielkonzentration erforderliche Volumen mit 5 mM Salzsäure mit 80 μg/mL Zink aufgefüllt. Falls erforderlich, wurde dabei der pH-Wert auf 3,5 + 0,1 nachjustiert. Nach der endgültigen Analyse durch HPLC zur Absicherung der Zielkonzentration von 240 ± 5 μM wurde die fertige Lösung mittels einer Spritze mit einem 0,2 μm Filtervorsatz in ein mit einem Septum und einer Bördelkappe verschlossenes steriles Fläschchen überführt. Für die kurzfristige, einmalige Testung der erfindungsgemäßen Insulinderivate wurde keine Optimierung der Formulierungen, z.B. hinsichtlich eines Zusatzes isotonischer Agentien, Konservierungsmittel oder Puffersubstanzen, vorgenommen. (Example 9) and then appropriate tests were made (Examples 10 and 11). The insulin analog according to the invention was dissolved at a target concentration of 240 + 5 μM in 1 mM hydrochloric acid with 80 μg / ml zinc (as zinc chloride). For this purpose, of the freeze-dried material was first weighed about 30% higher amount than required due to the molecular weight and the desired concentration. Thereafter, the present concentration was determined by means of analytical HPLC and the solution was then filled to the required volume to achieve the target concentration with 5 mM hydrochloric acid with 80 ug / mL zinc. If necessary, the pH was readjusted to 3.5 + 0.1. After final analysis by HPLC to ensure the target concentration of 240 ± 5 μM, the final solution was transferred via syringe with a 0.2 μm filter attachment to a sterile vial sealed with a septum and crimp cap. For the short-term, one-time testing of the insulin derivatives according to the invention, no optimization of the formulations, eg with regard to an addition of isotonic agents, preservatives or buffer substances, was carried out.
Beispiel 10: Evaluierung der blutzuckersenkenden Wirkung von neuen Insulinanaloga in der Ratte Die blutzuckersenkende Wirkung von ausgewählten neuen Insulinanaloga wird in männlichen, gesunden, normoglykämischen Wistarratten geprüft. Männlichen Ratten wird eine Dosis von 9 nmol/kg eines Insulinanalogons subkutan injiziert. Unmittelbar vor der Injektion des Insulinanalogons und in regelmäßigen Abständen bis zu acht Stunden nach der Injektion werden den Tieren Blutproben entnommen und darin der Blutzuckergehalt bestimmt. Das Experiment zeigt deutlich (vgl. Fig. 4), dass das erfindungsgemäße Insulinanalogon zu einem deutlich verzögerten Wirkeintritt und einer längeren, gleichmäßigen Wirkdauer führt. Example 10: Evaluation of the hypoglycemic effect of novel insulin analogues in the rat The hypoglycemic effect of selected new insulin analogs is tested in male, normal, normoglycemic Wistar rats. Male rats are injected subcutaneously with a dose of 9 nmol / kg of an insulin analogue. Immediately before injecting the insulin analogue and periodically up to eight hours after the injection, blood samples are taken from the animals and the blood sugar content is determined therein. The experiment clearly shows (compare Fig. 4) that the insulin analog according to the invention leads to a significantly delayed onset of action and a longer, uniform duration of action.
Beispiel 11 : Evaluierung der blutzuckersenkenden Wirkung von neuen Insulinanaloga im Hund Die blutzuckersenkende Wirkung von ausgewählten neuen Insulinanaloga wird in männlichen, gesunden, normoglykämischen Beaglehunden geprüft. Männlichen Tieren wird eine Dosis von 6 nmol/kg eines Insulinanalogons subkutan injiziert. Unmittelbar vor der Injektion des Insulinanalogons und in regelmäßigen Abständen bis zu achtundvierzig Stunden nach der Injektion werden den Tieren Blutproben entnommen und darin der Blutzuckergehalt bestimmt. Das Experiment zeigt deutlich, dass das erfindungsgemäße Insulinanalogon zu einem deutlich verzögerten, flachen Wirkeintritt und einer längeren, gleichmäßigen Wirkdauer führt. Example 11: Evaluation of the hypoglycemic effect of new insulin analogues in the dog The hypoglycemic effect of selected new insulin analogues is tested in male, healthy, normoglycemic beagle dogs. Male animals are injected subcutaneously with a dose of 6 nmol / kg of an insulin analogue. Immediately before injecting the insulin analogue and at regular intervals up to forty-eight hours after the injection, blood samples are taken from the animals and the blood sugar content is determined therein. The experiment clearly shows that the insulin analog according to the invention leads to a markedly delayed, shallow onset of action and a longer, uniform duration of action.

Claims

Patentansprüche: claims:
1. Wässrige pharmazeutische Formulierungen enthaltend ein Insulinanalogon der Formel I s c 1. Aqueous pharmaceutical formulations containing an insulin analog of the formula I s c
1 5 | 101 15 2u  1 5 | 101 15 2u
AO G I V E A5 C C H S I C S L Y A15 L E A18 Y C G  AO G I V E A5 C C H S I C S L Y A15 L E A18 Y C G
(SEQ ID NO: I) \ A-Kette (SEQ ID NO: I) \ A chain
S x S S x S
-S S -S S
I I  I i
B-I BO Bl B2 B3 B4 H L C G S H L V E A L Y L V C G E R G F F Y B-I BO B1 B2 B3 B4 H L C G S H L V E A L Y L V C G E R G F F Y
1 5 10 15 20 25 1 5 10 15 20 25
T P B29 B30 B31 B32 (SEQ ID NO: 2) T P B29 B30 B31 B32 (SEQ ID NO: 2)
B-Kette B chain
30 wobei  30 where
AO Lys oder Arg; AO Lys or Arg;
A5 Asp, GIn oder GIu; A5 Asp, GIn or GIu;
A15 Asp, GIu oder GIn; A18 Asp, GIu oder Asn; A15 Asp, Glu or GIn; A18 Asp, Glu or Asn;
B- 1 Asp, GIu oder eine Aminogruppe; B-1 Asp, Glu or an amino group;
BO Asp, GIu oder eine chemische Bindung; BO Asp, Glu or a chemical bond;
B1 Asp, GIu oder Phe; B2 Asp, GIu oder VaI; B1 Asp, Glu or Phe; B2 Asp, Glu or VaI;
B3 Asp, GIu oder Asn; B4 Asp, GIu oder GIn; B3 Asp, Glu or Asn; B4 Asp, Glu or GIn;
B29 Lys oder einer chemischen Bindung; B29 Lys or a chemical bond;
B30 Thr oder einer chemischen Bindung; B30 Thr or a chemical bond;
B31 Arg, Lys oder einer chemischen Bindung; B31 Arg, Lys or a chemical bond;
B32 Arg-Amid, Lys-Amid oder einer Aminogruppe entspricht, wobei zwei Aminosäurereste der Gruppe enthaltend A5, A15, A18, B-1 , BO, B1 , B2, B3 und B4 gleichzeitig und unabhängig voneinander Asp oder GIu B32 Arg amide, Lys amide or an amino group, wherein two amino acid residues of the group containing A5, A15, A18, B-1, BO, B1, B2, B3 and B4 are simultaneously and independently Asp or GIu
entsprechen, oder ein pharmakologisch tolerierbares Salz davon; und die or a pharmacologically tolerable salt thereof; and the
0,001 bis 0,2 mg/ml Zink, From 0.001 to 0.2 mg / ml of zinc,
0,1 bis 5,0 mg/ml eines Konservierungsmittels und 0.1 to 5.0 mg / ml of a preservative and
5,0 bis 100 mg/ml eines Isotonisierungsmittels enthält und  Contains 5.0 to 100 mg / ml of an isotonizing agent and
deren pH-Wert 5 oder kleiner ist. whose pH is 5 or less.
2. Pharmazeutische Formulierung gemäß Anspruch 1 , bei der das Insulinanalogon ausgewählt ist aus einer Gruppe enthaltend: 2. A pharmaceutical formulation according to claim 1, wherein the insulin analog is selected from a group comprising:
Arg (AO), His (A8), GIu (A5), Asp (A18), GIy (A21), Arg (B31), Arg (B32) - NH2 Humaninsuiin, Arg (AO), His (A8), GIu (A5), Asp (A18), GIy (A21 ), Arg (B31), Lys (B32) - NH2 Humaninsuiin, Arg (AO), His (A8), GIu (A15), Asp (A18), GIy (A21), Arg (B31 ), Arg (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIu (A15), Asp (A18), GIy (A21), Arg (B31), Lys (B32) - NH2 Humaninsulin, Arg (AO)1 His (A8), Glu(A5), GIu (A15), GIy (A21), Arg (B31), Arg (B32) - NH2 Humaninsulin, Arg (AO), His (A8), Giu (A5), GIu (A15), GIy (A21), Arg (B31 ), Lys (B32) - NH2 Humaninsuiin, Arg (AO), His(A8), GIu (A5), GIy (A21), Asp (B3), Arg (B31), Arg (B32) - NH2 Humaninsulin, Arg (AO), His(A8), GIu (A5), GIy (A21), Asp (B3), Arg (B31), Lys (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIu (A15), GIy (A21), Asp (B3), Arg (B31), Arg (B32)- NH2Humaninsuiin( Arg (AO), His (A8), GIu (A15), GIy (A21), Asp (B3), Arg (B31), Lys (B32)- NH2Humaninsuiin( Arg (AO)1 His (A8), Asp (A18), GIy (A21), Asp (B3), Arg (B31), Arg (B32)- NH2 Humaninsulin, Arg (AO), His (A8), Asp (A18), GIy (A21), Asp (B3), Arg (B31), Lys (B32)- NH2 Humaninsulin, Arg (AO), His(A8), GIy (A21 ), Asp (B3), GIu (B4), Arg (B31 ), Arg (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIy (A21), Asp (B3), GIu (B4), Arg (B31), Lys (B32)- NH2 Humaninsulin, Arg (AO), His (A8), GIu (A5), GIy (A21), GIu (B4), Arg (B31), Arg (B32)- NH2 Humaninsulin, Arg (AO), His (A8), GIu (A5), GIy (A21), GIu (B4), Arg (B31), Lys (B32)- NH2 Humaninsulin, Arg (AO), His (A8), GIu (A15), GIy (A21 ), GIu (B4), Arg (B31 ), Arg (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIu (A15), GIy (A21 ), GIu (B4), Arg (B31 ), Lys (B32) - NH2 Humaninsuiin, Arg (AO), His (A8), Asp (A18), GIy (A21), GIu (B4), Arg (B31), Arg (B32)- NH2 Humaninsulin, Arg (AO), His (A8), Asp (A18), GIy (A21), GIu (B4), Arg (B31), Lys (B32) - NH2 Humaninsulin, Arg (AO)1 His (A8), GIu (A5), GIy (A21 ), GIu (BO), Arg (B31), Arg (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIu (A5), GIy (A21), GIu (BO), Arg (B31), Lys (B32)- NH2 Humaninsulin, Arg (AO), His (A8), GIu (A15), GIy (A21), Giu (BO), Arg (B31), Arg (B32)- NH2 Humaninsuiin, Arg (AO), His (A8), GIu (A15), GIy (A21), GIu (BO), Arg (B31 ), Lys (B32) - NH2 Humaninsulin, Arg (AO), His (A8), Asp (A18), GIy (A21), GIu (BO), Arg (B31), Arg (B32) - NH2 Humaninsulin, Arg (AO), His (A8), Asp (A18) ,GIy (A21), Giu (BO), Arg (B31), Lys (B32)- NH2 Humaninsuiin, Arg (AO), His (A8), GIu (A5), GIy (A21), Asp (B1), Arg (B31), Arg (B32)- NH2 Humaninsulin, Arg (AO), His (A8), GIu (A5), Giy (A21 ), Asp (B1 ), Arg (B31 ), Lys (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIu (A15), GIy (A21), Asp (B1), Arg (B31), Arg(B32)- NH2 Humaninsulin, Arg (AO), His (A8), GIu (A15), GIy (A21), Asp (B1), Arg (B31), Lys (B32)- NH2 Humaninsuiin, Arg (AO), His (A8), Asp (A18), GIy (A21), Asp (B1), Arg (B31), Arg (B32)- NH2 Humaninsuiin, Arg (AO), His (A8), Asp (A18), GIy (A21), Asp (B1), Arg (B31), Lys (B32)- NH2 Humaninsuiin, Arg (AO), His (A8), GIy (A21 ), GIu (BO), Asp (B1 ), Arg (B31 ), Arg (B32) - NH2 Humaninsulin, Arg (AO), His (A8), GIy (A21 ), GIu (BO), Asp (B1 ), Arg (B31), Lys (B32) - NH2 Humaninsulin, Arg (AO), His (A8), Asp (A18), GIy (A21), Asp (B3), Arg (B30), Arg (B31)- NH2 Humaninsuiin, Arg (AO), His (A8), Asp (A18), GIy (A21), Asp (B3), Arg (B30), Lys (B31)- NH2 Humaninsulin. Arg (AO), His (A8), Glu (A5), Asp (A18), Gly (A21), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5), Asp (A18), Gly (A21), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A15), Asp (A18), Gly ( A21), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A15), Asp (A18), Gly (A21), Arg (B31), Lys (B32 ) - NH 2 human insulin, Arg (AO) 1 His (A8), Glu (A5), Glu (A15), Gly (A21), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Giu (A5), Glu (A15), Gly (A21), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5), Gly (A21), Asp (B3), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5), Gly (A21), Asp (B3), Arg (A21) B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A15), Gly (A21), Asp (B3), Arg (B31), Arg (B32) - NH 2 Human insulin ( Arg (AO), His (A8), Glu (A15), Gly (A21), Asp (B3), Arg (B31), Lys (B32) - NH 2 human insulin ( Arg (AO) 1 His (A8), Asp (A18), Gly (A21), Asp (A) B3), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Asp (A18), Gly (A21), Asp (B3), Arg (B31), Lys (B32 ) - NH 2 human insulin, Arg (AO), His (A8), Gly (A21), Asp (B3), Glu (B4), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Gly (A21), Asp (B3), Glu (B4), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5), Gly (A21), Glu (B4), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5), Gly (A21), Glu (B4), Arg (A21) B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A15), Gly (A21), Glu (B4), Arg (B31), Arg (B32) - NH 2 human insulin , Arg (AO), His (A8), Glu (A15), Gly (A21), Glu (B4), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Asp (A18), Gly (A21), Glu (B4), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Asp (A18), Gly (A21), Glu (B4), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (A) AO) 1 His (A8), Glu (A5), Gly (A21), Glu (BO), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5 ), Gly (A21), Glu (BO), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A15), Gly (A21), Giu (BO) , Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A15), Gly (A21), Glu (BO), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Asp (A18), Gly (A21), Glu (BO), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His ( A8), Asp (A18), Gly (A21), Giu (BO), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5), Gly (A21 ), Asp (B1), Arg (B31) Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A5), Gly (A21), Asp (B1), Arg (B31) , Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A15), Gly (A21), Asp (B1), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Glu (A15), Gly (A21), Asp (B1), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Asp (A18), Gly (A21), Asp (B1), Arg (A21) B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Asp (A18), Gly (A21), Asp (B1), Arg (B31), Lys (B32) - NH 2 human insulin , Arg (AO), His (A8), Gly (A21), Glu (BO), Asp (B1), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), His (A8), Gly (A21), Glu (BO), Asp (B1), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Asp (A18), Gly (A21), Asp (B3), Arg (B30), Arg (B31) - NH 2 Humaninsuiin, Arg (AO), His (A8), Asp (A18), Gly (A21), Asp (B3), Arg (B30), Lys ( B31) - NH 2 human insulin.
3. Pharmazeutische Formulierung gemäß einem der Ansprüche 1 oder 2, wobei das Konservierungsmittel ausgewählt wird aus einer Gruppe enthaltend Phenol, m-Cresol, Chlorkresol, Benzylalkohol, Parabene. 3. A pharmaceutical formulation according to any one of claims 1 or 2, wherein the preservative is selected from a group containing phenol, m-cresol, chlorocresol, benzyl alcohol, parabens.
4. Pharmazeutische Formulierung gemäß einem der Ansprüche 1 bis 3, wobei das Isotonisierungsmittel ausgewählt wird aus einer Gruppe enthaltend Mannitol, Sorbitol, Lactose, Dextrose, Trehalose, Natriumchlorid, Glycerol. 5. Pharmazeutische Formulierung gemäß einem der Ansprüche 1 bis 4, mit einem pH- Wert im Bereich von pH 2,5 - 4,4. A pharmaceutical formulation according to any one of claims 1 to 3, wherein the isotonizing agent is selected from a group containing mannitol, sorbitol, lactose, dextrose, trehalose, sodium chloride, glycerol. 5. A pharmaceutical formulation according to any one of claims 1 to 4, having a pH in the range of pH 2.5-4,
5. 5th
6. Pharmazeutische Formulierung gemäß einem der Ansprüche 1 bis 5, mit einem pH- Wert im Bereich von pH 3,0 - 4,0. 6. A pharmaceutical formulation according to any one of claims 1 to 5, having a pH in the range of pH 3.0-4.0.
7. Pharmazeutische Formulierung gemäß einem der Ansprüche 1 bis 6, mit einem pH- Wert im Bereich von pH 3,75. 7. A pharmaceutical formulation according to any one of claims 1 to 6, having a pH in the range of pH 3.75.
8. Pharmazeutische Formulierung gemäß einem der Ansprüche 1 bis 7, wobei das Insulin, das Insulinanalogon und/oder das Insulinderivat in einer Konzentration von 240 - 3000 nmol/ml vorliegt 8. A pharmaceutical formulation according to any one of claims 1 to 7, wherein the insulin, the insulin analogue and / or the insulin derivative is present in a concentration of 240-3000 nmol / ml
9. Pharmazeutische Formulierung gemäß einem oder mehreren der Ansprüche 1 bis 8, bei der Glycerol in einer Konzentration von 20 bis 30 mg/ml vorliegt. 9. Pharmaceutical formulation according to one or more of claims 1 to 8, in which glycerol is present in a concentration of 20 to 30 mg / ml.
10. Pharmazeutische Formulierung gemäß einem oder mehreren der Ansprüche 1 bis 8, bei der Glycerol in einer Konzentration von 25 mg/ml vorliegt. 10. Pharmaceutical formulation according to one or more of claims 1 to 8, in which glycerol is present in a concentration of 25 mg / ml.
11. Pharmazeutische Formulierung gemäß einem oder mehreren der Ansprüche 1 bis 10, bei der m-Cresol in einer Konzentration von 1 bis 3 mg/ml vorliegt. 11. Pharmaceutical formulation according to one or more of claims 1 to 10, wherein m-cresol is present in a concentration of 1 to 3 mg / ml.
12. Pharmazeutische Formulierung gemäß einem oder mehreren der Ansprüche 1 bis12. Pharmaceutical formulation according to one or more of claims 1 to
11 , bei der m-Cresol in einer Konzentration von 2 mg/ml vorliegt. 11, in which m-cresol is present at a concentration of 2 mg / ml.
13. Pharmazeutische Formulierung gemäß einem oder mehreren der Ansprüche 1 bis13. Pharmaceutical formulation according to one or more of claims 1 to
12, bei der Zink in einer Konzentration 0,01 oder 0,03 oder 0,08 mg/ml vorliegt. 12, in which zinc is present in a concentration of 0.01 or 0.03 or 0.08 mg / ml.
14. Pharmazeutische Formulierung nach einem oder mehreren der Ansprüche 1 bis 13, bei der noch zusätzlich ein Glucagon-Like Peptide-1 (GLP1 ) oder ein Analogon oder Derivat davon, oder Exendin-3 bzw. -4 oder ein Analogon oder Derivat davon enthalten ist. 14. Pharmaceutical formulation according to claim 1, further comprising a glucagon-like peptide-1 (GLP1) or an analogue or derivative thereof, or exendin-3 or -4 or an analogue or derivative thereof is.
15. Pharmazeutische Formulierung nach Anspruch 14, bei dem zusätzlich Exendin-4 enthalten ist. 15. A pharmaceutical formulation according to claim 14, which additionally contains exendin-4.
16. Pharmazeutische Formulierung gemäß Anspruch 14, bei dem ein Analogon von Exendin-4 ausgewählt wird aus einer Gruppe enthaltend A pharmaceutical formulation according to claim 14, wherein an analog of exendin-4 is selected from a group comprising
H-desPro3δ-Exendin-4-Lys6-NH2, H-desPro 3δ -exendin-4-Lys 6 -NH 2 ,
H-des(Pro3637)-Exendin-4-Lys4-NH2 und H-des (Pro 3637 ) -Exendin-4-Lys 4 -NH 2 and
H-des(Pro36'37)-Exendin-4-I_ys5-NH2, H des (Pro 36 '37 ) -Exendin-4-I_ys 5 -NH 2 ,
oder ein pharmakologisch tolerierbares Salz davon. or a pharmacologically tolerable salt thereof.
17. Pharmazeutische Formulierung gemäß Anspruch 14, bei dem ein Analogon von Exendin-4 ausgewählt wird aus einer Gruppe enthaltend 17. A pharmaceutical formulation according to claim 14, wherein an analog of exendin-4 is selected from a group comprising
desPro36 [Asp28jExendin-4 (1-39), desPro 36 [Asp 28 jExendin-4 (1-39),
desPro36 [lsoAsp2δ]Exendin-4 (1-39), desPro 36 [lsoAsp 2δ ] exendin-4 (1-39),
desPro36 [Met(O)14, Asp28]Exendin-4 (1-39), DESpro 36 [Met (O) 14, Asp 28] Exendin-4 (1-39),
desPro36 [Met(O)14, lsoAsp2δjExendin-4 (1-39), desPro 36 [Met (O) 14 , Isosp 2δ jExendin-4 (1-39),
desPro36 [Trp(O2)25, Asp28jExendin-2 (1-39), desPro36 [Trp(O2)25, lsoAsp28]Exendin-2 (1-39), desPro 36 [Trp (O 2 ) 25 , Asp 28, jExendin-2 (1-39), Pro 36 [Trp (O 2 ) 25 , IsoSp 28 ], Exendin-2 (1-39),
desPro36 [Met(O)14Trp(O2)25, Asp28JExendin-4 (1 -39) und desPro 36 [Met (O) 14 Trp (O 2 ) 25 , Asp 28 JExendin-4 (1 -39) and
desPro36 [Met(O)14Trp(O2)25, lsoAsp28]Exendin-4 (1-39), desPro 36 [Met (O) 14 Trp (O 2 ) 25 , IsoPs 28 ] Exendin-4 (1-39),
oder ein pharmakologisch tolerierbares Salz davon. or a pharmacologically tolerable salt thereof.
18. Pharmazeutische Formulierung gemäß Anspruch 17, bei denen an die C-Termini der Analoga von Exendin-4 das Peptid -Lys6-NH2 angefügt ist. A pharmaceutical formulation according to claim 17, wherein the peptide -Lys 6 -NH 2 is added to the C-termini of the analogues of exendin-4.
19. Pharmazeutische Formulierung gemäß Anspruch 14, bei dem ein Analogon von Exendin-4 ausgewählt wird aus einer Gruppe enthaltend 19. A pharmaceutical formulation according to claim 14, wherein an analog of exendin-4 is selected from a group comprising
H-(LyS)6- des Pro36 [Asp28]Exendin-4(1-39)-Lys6-NH2 H- (LyS) 6 - of Pro 36 [Asp 28 ] Exendin-4 (1-39) -Ly 6 -NH 2
des Asp28Pro3δ, Pro37, Pro38 Exendin-4(1-39) -NH2, Asp 28 Pro 3δ , Pro 37 , Pro 38 Exendin-4 (1-39) -NH 2 ,
H-(LyS)6- des Pro36, Pro37, Pro38 [Asp28]Exendin-4(1-39) -NH2, H- (LyS) 6 - of Pro 36 , Pro 37 , Pro 38 [Asp 28 ] Exendin-4 (1-39) -NH 2 ,
H-Asn-(Glu)5 des Pro36, Pro37, Pro38 [Asp28jExendin-4(1-39) -NH2, H-Asn- (Glu) 5 of the Pro 36 , Pro 37 , Pro 38 [Asp 28 jExendin-4 (1-39) -NH 2 ,
des Pro36, Pro37, Pro38 [Asρ28]Exendin-4(1 -39)-(Lys)6-NH2, Pro 36 , Pro 37 , Pro 38 [Asρ 28 ] Exendin-4 (1 -39) - (Lys) 6 -NH 2 ,
H-(LyS)6- des Pro36, Pro37, Pro38 [Asp28jExendin-4(1-39)-(Lys)6-NH2, H- (LyS) 6 - of Pro 36 , Pro 37 , Pro 38 [Asp 28 jExendin-4 (1-39) - (Lys) 6 -NH 2 ,
H-Asn-(Glu)5- des Pro36, Pro37, Pro38 [Asp28]Exendin-4(1 -39)-(Lys)6-NH2, H-Asn- (Glu) 5 - of the Pro 36 , Pro 37 , Pro 38 [Asp 28 ] Exendin-4 (1 -39) - (Lys) 6 -NH 2 ,
H-(LyS)6- des Pro36 [Trp(O2)25, Asp28]Exendin-4(1-39)-Lys6-NH2) H- (LyS) 6 - of Pro 36 [Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1-39) -Lys 6 -NH 2)
H- des Asp28 Pro36, Pro37, Pro38 [Trp(O2)25]Exendin-4(1-39) -NH2, H- the Asp 28 Pro 36, Pro 37, Pro 38 [Trp (O2) 25] Exendin-4 (1-39) -NH 2,
H-(LyS)6- des Pro36, Pro37, Pro38 [Trp(O2)25, Asp28jExendin-4(1-39) -NH2, H- (LyS) 6 - of Pro 36 , Pro 37 , Pro 38 [Trp (O 2 ) 25 , Asp 28 jExendin-4 (1-39) -NH 2 ,
H-Asn-(Glu)5- des Pro36, Pro37, Pro38 [Trp(O2)25, Asp28]Exendin-4(1-39) -NH2, des Pro36, Pro37, Pro38 [Trp(O2)25, Asp28]Exendin-4(1-39)-(Lys)6-NH2, H-Asn- (Glu) 5 - of Pro 36 , Pro 37 , Pro 38 [Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1-39) -NH 2 , of Pro 36 , Pro 37 , Pro 38 [Trp (O 2 ) 25 , Asp 28 ] exendin-4 (1-39) - (Lys) 6 -NH 2 ,
H-(LyS)6- des Pro36, Pro37, Pro38 [Trp(O2)25, Asp28jExendin-4(1-39)-(Lys)6-NH2, H-Asn-(Glu)5- des Pro36, Pro37, Pro38 [Trp(O2)25, Asp28]Exendin-4(1-39)-(Lys)6-NH2, H-(LyS)6- des Pro36 [Met(O)14, Asp28]Exendin-4(1-39)-Lys6-NH2, H- (LyS) 6 - of the Pro 36 , Pro 37 , Pro 38 [Trp (O 2 ) 25 , Asp 28, jExendin-4 (1-39) - (Lys) 6 -NH 2 , H-Asn- (Glu) 5 - of the Pro 36 , Pro 37 , Pro 38 [Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1-39) - (Lys) 6 -NH 2 , H- (LyS) 6 - of the Pro 36 [ Met (O) 14 , Asp 28 ] Exendin-4 (1-39) -Lys 6 -NH 2 ,
des Met(O)14 Asp28 Pro 36, Pro37, Pro38 Exendin-4(1-39) -NH2, the Met (O) 14 Asp 28 Pro 36, Pro 37, Pro 38 exendin-4 (1-39) -NH 2,
H-(LyS)6- des Pro36, Pro 37, Pro38 [Met(O)14, Asp28]Exendin-4(1-39) -NH2, H- (LyS) 6 - of Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Asp 28 ] Exendin-4 (1-39) -NH 2 ,
H-ASn-(GIu)5- des Pro36, Pro37, Pro38 [Met(O)14, Asp28] Exendin-4(1 -39) -NH2, des Pro36, Pro37, Pro38 [Met(O)14, Asp28]Exendin-4(1-39)-(Lys)6-NH2, H-ASn (Glu) 5 - of Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Asp 28 ] Exendin-4 (1 -39) -NH 2 , of Pro 36 , Pro 37 , Pro 38 [ Met (O) 14 , Asp 28 ] Exendin-4 (1-39) - (Lys) 6 -NH 2 ,
H-(LyS)6- des Pro36, Pro37, Pro38 [Met(O)14, Asp28]Exendin-4(1-39)-Lys6-NH2, H-Asn-(Glu)5 des Pro36, Pro37, Pro38 [Met(O)14, Asp28] Exendin-4(1-39)-(Lys)6-NH2, H- (LyS) 6 - of Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Asp 28 ] Exendin-4 (1-39) -Lys 6 -NH 2 , H-Asn- (Glu) 5 of Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Asp 28 ] Exendin-4 (1-39) - (Lys) 6 -NH 2 ,
H-(LyS)6- des Pro36 [Met(O)14, Trp(O2)25, Asp28]Exendin-4(1-39)-Lys6-NH2, des Asp28 Pro36, Pro37, Pro38 [Met(O)14, Trp(O2)25]Exendin-4(1-39) -NH2, H- (LyS) 6 - of Pro 36 [Met (O) 14 , Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1-39) -Lys 6 -NH 2 , of Asp 28 Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Trp (O 2 ) 25 ] Exendin-4 (1-39) -NH 2 ,
H-(LyS)6- des Pro36' Pro37, Pro38 [Met(O)14, Trp(O2)25, Asp28]Exendin-4(1-39) -NH2, H-Asn-(Glu)5- des Pro36, Pro37, Pro38 [Met(O)14, Asp28] Exendin-4(1-39) -NH2, des Pro36, Pro37, Pro38 [Met(O)14, Trp(O2)25, Asp2δ]Exendin-4(1-39)-(Lys)6-NH2,H- (LyS) 6 - of Pro 36 'Pro 37 , Pro 38 [Met (O) 14 , Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1-39) -NH 2 , H-Asn- ( Glu) 5 - Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Asp 28 ] Exendin-4 (1-39) -NH 2 , Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Trp (O 2 ) 25 , Asp 2δ ] exendin-4 (1-39) - (Lys) 6 -NH 2 ,
H-(LyS)6- des Pro36' Pro37, Pro38 [Met(O)14, Trp(O2)25, Asp28]Exendin-4(1 -39)-(Lys)6- NH2, H-ASn-(GIu)5- des Pro36, Pro37, Pro38 [Met(O)14, Trp(O2)25, Asp28] Exendin-4(1-39)-H- (LyS) 6 - of Pro 36 'Pro 37 , Pro 38 [Met (O) 14 , Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1 -39) - (Lys) 6 - NH 2 , H-ASn- (Glu) 5 - of Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Trp (O 2 ) 25 , Asp 28 ] Exendin-4 (1-39) -
(LyS)6-NH2, (LyS) 6 -NH 2 ,
oder ein pharmakologisch tolerierbares Salz davon. or a pharmacologically tolerable salt thereof.
20. Pharmazeutische Formulierung nach Anspruch 14, bei dem zusätzlich Arg34, Lys26 (Nε(γ-glutamyl(Nα-hexadecanoyl))) GLP-1 (7-37) [liraglutide] oder ein pharmakologisch tolerierbares Salz davon enthalten ist. A pharmaceutical formulation according to claim 14 further comprising Arg 34 , Lys 26 (N ε (γ-glutamyl (N α -hexadecanoyl)), GLP-1 (7-37) [liraglutide] or a pharmacologically tolerable salt thereof ,
21. Pharmazeutische Formulierung gemäß einem oder mehreren der Ansprüche 1 bis 20, bei der die Aminosäure Methionin vorhanden ist. 21. A pharmaceutical formulation according to one or more of claims 1 to 20, wherein the amino acid methionine is present.
22. Pharmazeutische Formulierung nach Anspruch 21 , bei der Methionin im 22. A pharmaceutical formulation according to claim 21, wherein the methionine im
Konzentrationsbereich von bis zu 10 mg/ml vorhanden ist. Concentration range of up to 10 mg / ml is present.
23. Pharmazeutische Formulierung nach Anspruch 22, bei der Methionin im 23. A pharmaceutical formulation according to claim 22, wherein the methionine im
Konzentrationsbereich von bis zu 3 mg/ml vorhanden ist. Concentration range of up to 3 mg / ml is present.
24. Verfahren zur Herstellung einer Formulierung gemäß einem oder mehreren der Ansprüche 1 bis 20, bei dem 24. A process for the preparation of a formulation according to one or more of claims 1 to 20, in which
(a) die Komponenten in eine wässrige Lösung eingebracht werden und (a) the components are introduced into an aqueous solution and
(b) der pH-Wert eingestellt wird. (b) the pH is adjusted.
25. Verwendung einer Formulierung gemäß einem oder mehreren der Ansprüche 1 bis 20 zur Behandlung von Diabetes mellitus. 25. Use of a formulation according to one or more of claims 1 to 20 for the treatment of diabetes mellitus.
26. Arzneimittel zur Behandlung von Diabetes mellitus bestehend aus einer 26. Medicaments for the treatment of diabetes mellitus consisting of a
Formulierung gemäß einem oder mehreren der Ansprüche 1 bis 20. A formulation according to one or more of claims 1 to 20.
EP10726993A 2009-07-06 2010-07-02 Slow-acting insulin preparations Withdrawn EP2451471A1 (en)

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