EP2451472A1 - Heat- and vibration-stable insulin preparations - Google Patents

Heat- and vibration-stable insulin preparations

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Publication number
EP2451472A1
EP2451472A1 EP10730441A EP10730441A EP2451472A1 EP 2451472 A1 EP2451472 A1 EP 2451472A1 EP 10730441 A EP10730441 A EP 10730441A EP 10730441 A EP10730441 A EP 10730441A EP 2451472 A1 EP2451472 A1 EP 2451472A1
Authority
EP
European Patent Office
Prior art keywords
arg
asp
insulin
glu
pro
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Withdrawn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
EP10730441A
Other languages
German (de)
French (fr)
Inventor
Anja Pfenninger
Norbert Tennagels
Christiane Fuerst
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 EP2451472A1 publication Critical patent/EP2451472A1/en
Withdrawn legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/28—Insulins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K47/02—Inorganic compounds
    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • 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

Definitions

  • the invention relates to a process for the preparation of an aqueous, pharmaceutical formulation containing an insulin, an insulin analog or an insulin derivative, wherein the ready-to-use formulation is obtained directly by dissolving the insulin, the
  • 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 can be life threatening in their most severe forms.
  • years of high blood glucose levels without initial symptoms represent a significant health risk.
  • the large-scale DCCT study in the USA The Diabetes Control and Complications Trial Research Group (1993) N. Engl. J. Med. 329, 977-986) clearly demonstrated that chronically elevated blood glucose levels are significantly responsible for the development of late diabetic lesions.
  • Diabetic late damage is microvascular and macrovascular damage, which may manifest as retinopathy, nephropathy, or neuropathy, leading to blindness, kidney failure, and loss of extremities, as well as an increased risk of cardiovascular disease
  • 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 obtained by chemical modification.
  • the chemical modification can eg in the addition of one or more certain consist of chemical groups to one or more amino acids.
  • insulin derivatives and insulin analogues have a slightly altered effect on human insulin. Insulin analogs with accelerated onset of action are described in EP 0 214 826,
  • 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
  • Liquid insulin preparations have a shelf life of approximately 2 years when stored at 2-8 ° C.
  • the shelf life in use allows storage at up to 25 ° C and is given with 4 weeks, mechanical stress (shaking) is too avoid.
  • mechanical stress shocking
  • patients must be careful to ensure that insulin preparation remains a clear solution, as in exceptional cases precipitation of insulin may occur, in part through the formation of so-called “fibrils", which may result in the risk of insufficient dosage of the medicine.
  • the 2-component insulin preparations of the present invention differ from conventional ones in their heat stability and mechanical shock resistance.
  • the heat and shake stability is based on the storage of insulin as a solid until shortly before administration. It has been shown that solid insulin is more stable to degradation (change of molecular structure) than to dissolved heat stress. Furthermore, dissolved insulin precipitates; this represents a biophysical process which can not take place in the solid state. As a result, with comparable heat stress, more bioavailable insulin is available to an insulin preparation when the dissolution process takes place after the heat stress.
  • Aqueous insulin preparations also show a tendency to precipitate insulin upon mechanical stress (shaking).
  • the amount of bioavailable insulin after shaking stress is thus unknown and represents an impairment of the
  • solvent surfactants
  • the solid insulin dissolution process may be performed on-site by the patient or his or her caregiver, for example in a two-chamber vial or other suitable device.
  • Corresponding attempts to completely dissolve insulins in the appropriate solvents and concentrations for an adequate duration ( ⁇ 10min) have been successful. Essentially, success depends on the appropriate pH of the solvent.
  • Solvents are not or not necessarily different from those already on the market. Heat and mech. Stress-resistant and insulins thus offer the following advantages:
  • the 2-component insulin preparations according to the invention offer the above-mentioned advantages. Patients with no or poor access to suitable refrigerators can thus keep appropriate amounts of insulin in stock; also is the
  • the amount of insulin solution in the case of patiens can also be reduced so that the necessary storage period in use can be reduced to a minimum. This may be the addition of antimicrobial
  • Additives such as m-cresol or other phenols are reduced or even obsolete.
  • the preparations can be used for all known insulins, insulin analogues and
  • Insulin derivatives are produced. These include preparations with desirable basal time /
  • the B chain end consists of an amidated basic amino acid residue such as lysine or
  • Argininamide exists, 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 one
  • 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 a process for the preparation of an aqueous, pharmaceutical formulation containing an insulin, an insulin analog or an insulin derivative, or a pharmacologically tolerable salt thereof, wherein the
  • Solvent mixture is carried out, preferably in which the composition of the suitable solvent mixture is determined by
  • Excipients are prepared which correspond to the final concentration of the excipients of the formulation containing an insulin, an insulin analog or an insulin derivative, and
  • Another object of the invention is a method as described above, wherein the insulin, the insulin analogue or the insulin derivative is present as a crystalline or amorphous solid.
  • An object of the invention is a method as described above, wherein the
  • Insulin is selected from a group containing human insulin, porcine insulin and bovine insulin.
  • the invention further provides a method as described above, wherein the insulin analog is selected from a group comprising Gly (A21), Arg (B31), Arg (B32) -human insulin, Lys (B3), Glu (B29) -human insulin, Asp (B28) human insulin, Lys (B28) Pro (B29) human insulin and Des (B30) human insulin.
  • Another object of the invention is a method according to one or more of claims 1 to 3, wherein the insulin analogue is selected from a group containing an insulin analog of the formula I.
  • insulin analog is selected from a group comprising:
  • Another object of the invention is a method as described above, wherein the insulin analogue is selected from a group containing
  • A1 Arg or Gly A5 Asp, Glu or GIn;
  • B-1 Asp Glu or an amino group
  • BO Asp Glu or a chemical bond
  • a further subject of the invention is a method according to one or more of claims 1 to 3, wherein the insulin derivative is selected from a group containing B29-N-myristoyl des (B30) human insulin, B29-N-palmitoyl des (B30) human insulin , B29-N-myhstoyl human insulin, B29-N-palmitoyl human insulin, B28-N-myristoyl Lys B28 Pro B29 human insulin, B28-N-palmitoyl-Lys B28 Pro B29 human insulin, B30-N-myhstoyl-Thr B29 Lys B3 ° human insulin , B30-N-palmitoyl-Thr B29 Lys B30
  • Another object of the invention is a method as described above, wherein in the formulation a preservative selected from a group containing phenol, m-cresol, chlorocresol, benzyl alcohol, parabens is present.
  • Another object of the invention is a method as described above, wherein in the formulation an isotonizing agent selected from a group containing mannitol, sorbitol, lactose, dextrose, trehalose, sodium chloride, glycerol is present.
  • Another object of the invention is a method as described above, wherein the insulin, the insulin analogue and / or the insulin derivative is present in a concentration of 240-3000 nmol / ml
  • Another object of the invention is a method as described above, wherein in the formulation additionally a glucagon-like peptide-1 (GLP1) or an analog or derivative thereof, or exendin-3 or -4 or an analog or derivative thereof is included , preferably in which an analog of exendin-4 is selected from a group comprising
  • Another object of the invention is a method as described above, wherein an analog of exendin-4 is selected from a group containing the Pro 36 [Asp 28 ] exend in-4 (1 -39),
  • 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 method as described above, wherein in the formulation additionally 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 method as described above, wherein in the formulation nor a zinc salt is included.
  • Another object of the invention is the use of a method as described above in the large-scale production of an insulin, insulin analog or insulin derivative.
  • Another object of the invention is a two-part set of containers, in which one of the containers an insulin, an insulin analogue or an insulin derivative as a solid and the other container a solvent mixture of a certain pH with the final concentration of the excipients of a desired formulation of an insulin, a Insulin analogues or an insulin derivative; for heat and shake-stable storage of insulin, the insulin analog or the
  • Insulin derivative for later preparing a ready-to-use formulation by dissolving the solid in the solvent mixture as described above.
  • a further subject of the invention is a two-chamber injection system in which one chamber contains an insulin, an insulin analog or an insulin derivative as solid and the other chamber
  • formulation and “preparation” are used interchangeably.
  • Fig. 1 Hypoglycemic effect of new insulin analogues according to formula I in rats
  • 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
  • Fig. 5 Hypoglycemic effect of insulin analogues of the invention
  • Fig. 6 Hypoglycemic effect of insulin glargine in rats
  • Example 1 Simplified dissolution of insulins in one step (dissolution test)
  • Composition of the market formulation i. taking into account the concentration of all auxiliaries and additives and the pH.
  • Lantus ® auxiliaries and additives zinc chloride, m-cresol, glycerin, pH 4
  • Insuman ® auxiliaries and additives Nathumhydrogenphosphat, m-cresol, glycerin, pH 7.3
  • Dissolution rate (final volume 1 mL) was less than 10 minutes.
  • Insuman ® required a pH of 7.6 to reach a complete solution with the required final pH of 7.3.
  • the dissolution process for the insulins could be realized within a reasonable time frame ( ⁇ 10 min).
  • additives contribute differently to the thermal stability of the dissolved insulin, e.g. the zinc concentration or the pH (N.R.
  • Containers for 14 days at 60 ° C.
  • sources production
  • containers from the same production batch were stored at 4 ° C. and analyzed together with the stressed samples.
  • Insulin preparations, heat- and shake-stable dosage form in a two-component system Large-scale production of insulin preparations:
  • Single-step resolution is a simplified process with the following advantages: fewer steps with fewer intermediate analyzes (e.g., pH) must be made, i.
  • the production of insulin preparations can be done faster.
  • the manufacture is less complicated, resulting in a simplified training of the employees (less complicated SOPs).
  • fewer containers are to be cleaned, which in turn saves working time and material.
  • Heat stress are more stable than dissolved. Degradation took place to a small extent, but loss by precipitation was eliminated, so that more bioavailable insulin was available with comparable heat stress if the dissolution process took place after the heat stress. Precipitation and degradation induced by vigorous shaking of the solid insulins could not be observed.
  • a presentation in solid form can thus be described as generally more temperature-stable and more robust to shaking and consequently safer.
  • administration of the insulins in solid form offers the additional advantage that accidental freezing by the patient can not lead to complication; Solid insulins are known to be stored in the frozen state.
  • Insulin powders instead of dissolved insulins are treated with suitable solvents (generally aqueous systems containing auxiliary agents and additives such as
  • solvents are offered in a two-component system
  • a two-chamber system is described, for example, in WO2007 / 038773 A1.
  • This increases the tolerable temperature range (both towards lower and higher temperatures).
  • the stability to mechanical stress such as strong vibrations increases. As a result, longer shelf lives, lower storage and transportation costs, and safer drug use are expected.
  • Simplified dissolution in one step ensures applicability by the patient.
  • 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). The following 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 novel insulin analogues in the rat The hypoglycemic effect of selected new insulin analogues 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 (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 it determines the blood sugar content.
  • the experiment clearly shows (see Fig. 2) that the insulin analog according to the invention used is a clear one
  • 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-response 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 Formulation of the amidated insulin derivatives
  • 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.
  • no optimization of the formulations e.g. with regard to an addition of isotonic agents, preservatives or buffer substances.
  • Example 10 Evaluation of the hypoglycemic effect of novel 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. 5) 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.
  • 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.

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Abstract

The invention relates to a method for producing an aqueous, pharmaceutical formulation comprising an insulin, an insulin analog, or an insulin derivative, wherein the ready-made formulation takes place directly by dissolving the insulin, the insulin analog, or the insulin derivative as a solid in a suitable solvent mixture.

Description

Beschreibung  description
Hitze- und schüttelstabile Insulinzubereitungen Die Erfindung betrifft ein Verfahren zur Herstellung einer wässrigen, pharmazeutischen Formulierung enthaltend ein Insulin, ein Insulinanalogon oder ein Insulinderivat, wobei die gebrauchsfertige Formulierung unmittelbar durch Lösen des Insulins, des The invention relates to a process for the preparation of an aqueous, pharmaceutical formulation containing an insulin, an insulin analog or an insulin derivative, wherein the ready-to-use formulation is obtained directly by dissolving the insulin, the
Insulinanalogons oder des Insulinderivats als Feststoff mit einem geeigneten Insulin analogues or the insulin derivative as a solid with a suitable
Lösemittelgemisch erfolgt. Solvent mixture takes place.
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 subcutane 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 Complications 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 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 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 can be life threatening in their most severe forms. In addition, however, years of high blood glucose levels without initial symptoms represent a significant health risk. The large-scale DCCT study in the USA (The Diabetes Control and Complications Trial Research Group (1993) N. Engl. J. Med. 329, 977-986) clearly demonstrated that chronically elevated blood glucose levels are significantly responsible for the development of late diabetic lesions. Diabetic late damage is microvascular and macrovascular damage, which may manifest as retinopathy, nephropathy, or neuropathy, leading to blindness, kidney failure, and loss of extremities, as well as 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. Insulinanaloga mit beschleunigtem Wirkungseintritt werden in EP 0 214 826, Insulin analog obtained by chemical modification. The chemical modification can eg in the addition of one or more certain consist of chemical groups to one or more amino acids. In general, insulin derivatives and insulin analogues have a slightly altered effect on human insulin. 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. Protected modifications are modified in the asparagine in B3 and at least one other amino acid in positions A5, A15, A18 or A21.
In der Regel haben Insulinderivate und Insulinanaloga gegenüber humanem Insulin eine etwas veränderte Wirkung. 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) 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 In general, insulin derivatives and insulin analogues have a slightly altered effect on human insulin. 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 medicine Lantus ® (active substance: insulin glargine = Gly (A21), Arg (B31), Arg (B32) 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 will certainly work in every patient for 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 by a delayed, ie 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)). Precipitations (particle formation) (Brange et al., J. Ph. Sei 86: 517-525 (1997)).
Flüssige Insulinzubereitungen haben bei einer Lagerung bei 2-8°C eine Haltbarkeitsdauer von ca. 2 Jahren. Die Haltbarkeit in Gebrauch erlaubt eine Lagerung bei bis zu 25°C und wird mit 4 Wochen angegeben, mechanischer Streß (Schütteln) ist zu vermeiden. Die Patienten müssen hierbei generell darauf achten, dass die Insulinzubereitung eine klare Lösung bleibt, da es in Ausnahmefällen zu Präzipitation des Insulins kommen kann, teilweise durch Bildung sogenannter„Fibrillen". Dadurch besteht die Gefahr, dass eine hinreichende Dosierung des Arzneimittels nicht gewährleistet ist. Liquid insulin preparations have a shelf life of approximately 2 years when stored at 2-8 ° C. The shelf life in use allows storage at up to 25 ° C and is given with 4 weeks, mechanical stress (shaking) is too avoid. In general, patients must be careful to ensure that insulin preparation remains a clear solution, as in exceptional cases precipitation of insulin may occur, in part through the formation of so-called "fibrils", which may result in the risk of insufficient dosage of the medicine.
Es besteht also der Bedarf, hitze- und schüttelstabilen stabile Insuline bzw. hitze- und schüttelstabilen stabile Insulinzubereitungen zur Behandlung des Typ 2 und Typ 1 Diabetes zu entwickeln. Dies ist im Rahmen der hier beschriebenen Erfindung erfolgt. Die erfindungsgemäßen 2-Komponenten-lnsulinzubereitungen unterscheiden sich von herkömmlichen durch ihre Hitzestabilität und ihrer Stabilität gegenüber mechanischem Steß. Im Gegensatz zu auf den Markt befindlichen flüssigen Insulinformulierungen beruht die Hitze- und Schüttelstabilität auf der Aufbewahrung von Insulin als Feststoff bis kurz vor der Darreichung. Es konnte gezeigt werden, daß festes Insulin bei Hitzestress gegenüber Degradation (Veränderung der Molekülstruktur) stabiler ist als gelöstes. Weiterhin präzipitiert gelöstes Insulin; dies stellt einen biophysikalischen Vorgang dar, welcher im festen Zustand nicht stattfinden kann. In der Folge steht bei vergleichbarem Hitzestress mehr bioverfügbares Insulin einer Insulinzubereitung zur Verfügung, wenn der Lösevorgang nach dem Hitzestress stattfindet. Thus, there is a need to develop stable insulins resistant to heat and shaking, and stable insulin preparations resistant to heat and shaking, for the treatment of type 2 and type 1 diabetes. This has been done within the scope of the invention described herein. The 2-component insulin preparations of the present invention differ from conventional ones in their heat stability and mechanical shock resistance. In contrast to liquid insulin formulations on the market, the heat and shake stability is based on the storage of insulin as a solid until shortly before administration. It has been shown that solid insulin is more stable to degradation (change of molecular structure) than to dissolved heat stress. Furthermore, dissolved insulin precipitates; this represents a biophysical process which can not take place in the solid state. As a result, with comparable heat stress, more bioavailable insulin is available to an insulin preparation when the dissolution process takes place after the heat stress.
Wäßrige Insulinzubereitungen zeigen weiterhin bei mechanischem Streß (Schütteln) die Neigung, Insulin zu präzipitieren. Die Menge des bioverfügbaren Insulins nach Schüttelstreß ist somit unbekannt und stellt eine Beeinträchtigung der Aqueous insulin preparations also show a tendency to precipitate insulin upon mechanical stress (shaking). The amount of bioavailable insulin after shaking stress is thus unknown and represents an impairment of the
Patientensicherheit dar. Zudem besteht somit außer hitzestabilen Insulinzubereitungen Bedarf an solchen, die auch gegenüber mechanischem Streß (z.B. Schütteln) stabil sind. Es konnte gezeigt werden, daß festes Insulin gegenüber Schütteln stabiler ist als gelöstes Insulin. Der biophysikalische Vorgang des Präzipitierens bei Schüttelstreß findet ausschließlich in gelöstem Insulin statt. In der Folge steht bei vergleichbarem Schüttelstreß bioverfügbares Insulin in unbeeinträchtiger Menge zur Verfügung, wenn der Lösevorgang nach dem Schüttelstreß stattfindet. Üblicherweise werden Insuline in wäßrigen Systemen, die Hilfs-und Zusatzstoffe wie beispielsweise Antibakterizide, isotonisierende Agenzien, Puffersubstanzen, und/oder Tenside enthalten (= im folgenden„Lösemittel" genannt) auf dem Markt angeboten. Das Auflösen von Insulinen zur Herstellung pharmakologischer Zubereitung wird im Allgemeinen dadurch gewährleistet, daß die Insuline zunächst sauer angelöst werden, um dann in weiteren Schritten auf die erforderliche Konzentration und pH-Wert der Lösung eingestellt zu werden (The Wellcome Foundation Limited te London, In addition to heat-stable insulin preparations there is thus a need for those which are also stable to mechanical stress (eg shaking). It has been shown that solid insulin is more stable to shaking than dissolved insulin. The biophysical process of precipitation on shaking occurs only in dissolved insulin. As a result, with comparable shaking stress, bioavailable insulin is available in an unremarkable amount if the dissolution process takes place after the shaking stress. Usually, insulins are available on the market in aqueous systems containing adjuvants and additives such as antibacterial agents, isotonizing agents, buffer substances, and / or surfactants (= "solvent") In general, it is ensured that the insulins are initially dissolved in acid, in order then to be adjusted in further steps to the required concentration and pH of the solution (The Wellcome Foundation Limited, London,
Octrooiraad Nederland, Octrooiannnvrage No. 6506714, 26.05.1965 "Werkwijze voor het bereiden van insulinepreparaten"). Octrooiraad Nederland, Octrooiannnvrage No. 6506714, 26.05.1965 "Werkwijze voor het bereiden van insulinepreparaten").
Alternativ ist ein Anlösen von Insulinen im Alkalischen und anschließender pH-Wert- Einstellung einhergehend mit einer leicht erhöhten Stabilität der Insulinzubereitungen beschrieben (WO 2004/096266). Somit ist jeweils ein mehrstufiger Prozeß zur Herstellung einer gebrauchsfertigen Insulinzubereitung nötig, der u.a. die Einstellung des pH-Werts umfasst. Alternatively, a solubilization of insulins in alkaline and subsequent pH adjustment is described along with a slightly increased stability of the insulin preparations (WO 2004/096266). Thus, in each case a multi-stage process for the preparation of a ready-insulin preparation is necessary, the u.a. the pH adjustment includes.
Es wurde nun überraschenderweise gefunden, dass sich gebrauchsfertige It has now surprisingly been found that ready to use
Insulinzubereitungen durch ein einstufiges Verfahren herstellen lassen, bei dem das Insulin (oder ein Analog oder Derivat davon) mit einer Pufferlösung Prepare insulin preparations by a one-step procedure in which the insulin (or analog or derivative thereof) is buffered
zusammengegeben wird und durch Auflösen des Insulins (oder des Analogs oder des Derivats davon) die gebrauchsfertige Insulinzubereitung innerhalb weniger Minuten entsteht. Somit kann das Auflöseverfahren des festen Insulins vor Ort durch den Patienten bzw. seiner betreuenden Personen erfolgen, beispielsweise in einer Zwei-Kammer-Ampulle oder anderen geeigneten Vorrichtungen. Entsprechende Versuche, Insuline in den geeigneten Lösemitteln und Konzentrationen in einer adäquaten Dauer (~10min) komplett zu lösen, sind erfolgreich verlaufen. Im wesentlichen hängt hierfür der Erfolg vom geeigneten pH-Wert des Lösemittels ab. Die prinzipielle Komposition der and, by dissolving the insulin (or the analog or derivative thereof), the ready-to-use insulin preparation is produced within a few minutes. Thus, the solid insulin dissolution process may be performed on-site by the patient or his or her caregiver, for example in a two-chamber vial or other suitable device. Corresponding attempts to completely dissolve insulins in the appropriate solvents and concentrations for an adequate duration (~ 10min) have been successful. Essentially, success depends on the appropriate pH of the solvent. The principal composition of
Lösemittel (Hilfsmittel, Zusatzstoffe etc.) unterscheidet sich dabei von den bereits auf dem Markt befindlichen Formulierungen nicht oder nicht notwendigerweise. Hitze- und mech. streßstabile und Insuline bieten somit folgende Vorteile:Solvents (auxiliaries, additives, etc.) are not or not necessarily different from those already on the market. Heat and mech. Stress-resistant and insulins thus offer the following advantages:
- Notwendige Kühlkette wird obsolet - Necessary cold chain becomes obsolete
- Lagervorhaltung in warmen Ländern vereinfacht  - Warehousing in warm countries simplified
- Verbesserte Arzneimittelsicherheit beim Patienten  - Improved drug safety in the patient
- Generelle verlängerte Haltbarkeitsdauer des Arzneimittels  - General prolonged shelf life of the drug
- Verringerter Rücklauf von Ampullen mit präzipitiertem Insulin  - Reduced return of ampoules of precipitated insulin
Die erfindungsgemäßen 2-Komponenten-lnsulinzubereitungen bieten oben genannte Vorteile. Patienten mit keinem oder mangelhaftem Zugang zu geeigneten Kühlgeräten können auf diese Weise geeignete Mengen Insulin vorrätig halten; auch ist die The 2-component insulin preparations according to the invention offer the above-mentioned advantages. Patients with no or poor access to suitable refrigerators can thus keep appropriate amounts of insulin in stock; also is the
Verwendung solcher Zubereitungen in Ländern mit warmem Klima besonders vorteilhaft. Durch geeignete Ampullengrößen im Sinne der erfindungsgemäßen 2-Komponenten- Insulinzubereitungen kann auch die Menge der Insulinlösung beim Patienen so verringert werden, daß die notwendige Lagerungsdauer in Gebrauch auf ein minimales Maß reduziert werden kann. Dadurch kann der Zusatz von antimikrobiellen Use of such preparations in countries with a warm climate particularly advantageous. By means of suitable ampule sizes in the sense of the 2-component insulin preparations according to the invention, the amount of insulin solution in the case of patiens can also be reduced so that the necessary storage period in use can be reduced to a minimum. This may be the addition of antimicrobial
Zusatzstoffen wie beispielsweise m-Kresol oder anderen Phenolen verringert oder sogar obsolet werden. Additives such as m-cresol or other phenols are reduced or even obsolete.
Die Zubereitungen können für alle bekannten Insuline, Insulinanaloga und The preparations can be used for all known insulins, insulin analogues and
Insulinderivate hergestellt werden. Dazu zählen auch Zubereitungen mit wünschenswerten basalen Zeit- / Insulin derivatives are produced. These include preparations with desirable basal time /
Wirkungsprofilen, bei denen die Insulinanaloga durch die Merkmale charakterisiert sind, dass  Activity profiles in which the insulin analogues are characterized by the features that
• das B- Kettenende aus einem amidierten basischen Aminosäurerest wie Lysin bzw. The B chain end consists of an amidated basic amino acid residue such as lysine or
Argininamid besteht, d.h. bei dem amidierten basischen Aminosäurerest am B- Kettenende liegt die Carboxylgruppe der endständigen Aminosäure in ihrer amidiertenArgininamide exists, 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 one
Form vor, und • der N-terminale Aminosäurerest der Insulin A-Kette ein Lysin- oder Argininrest ist, und Form before, and 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 und B4 erfolgt sind. Ein Gegenstand der Erfindung ist daher ein Verfahren zur Herstellung einer wässrigen, pharmazeutischen Formulierung enthaltend ein Insulin, ein Insulinanalogon oder ein Insulinderivat, oder ein pharmakologisch tolerierbares Salz davon, wobei die  Additions of negatively charged amino acid residues or such a substitution and such an addition in each case in the positions A5, A15, A18, B-1, BO, B1, B2, B3 and B4 are carried out. An object of the invention is therefore a process for the preparation of an aqueous, pharmaceutical formulation containing an insulin, an insulin analog or an insulin derivative, or a pharmacologically tolerable salt thereof, wherein the
gebrauchsfertige Formulierung unmittelbar durch Lösen des Insulins, des ready to use formulation immediately by dissolving the insulin, the
Insulinanalogons oder des Insulinderivats als Feststoff mit einem geeigneten Insulin analogues or the insulin derivative as a solid with a suitable
Lösemittelgemisch erfolgt, vorzugsweise bei dem die Zusammensetzung des geeigneten Lösemittelgemischs ermittelt wird, indem Solvent mixture is carried out, preferably in which the composition of the suitable solvent mixture is determined by
(a) Lösemittelgemische unterschiedlichen pH-Werts mit Konzentrationen von  (a) solvent mixtures of different pH with concentrations of
Exzipienten zubereitet werden, welche der Endkonzentration der Exzipienten der Formulierung enthaltend ein Insulin, ein Insulinanalogon oder ein Insulinderivat entsprechen und Excipients are prepared which correspond to the final concentration of the excipients of the formulation containing an insulin, an insulin analog or an insulin derivative, and
(b) durch Lösen des gewünschten Insulins, Insulinanalogons oder Insulinderivats dasjenige Lösemittelgemisch ermittelt wird, welches nach Lösen des Feststoffes von Insulin, Insulinanalogon oder Insulinderivat den gewünschten pH-Wert der  (B) by dissolving the desired insulin, insulin analogue or insulin derivative that solvent mixture is determined which, after dissolving the solid of insulin, insulin analogue or insulin derivative, the desired pH of the
gebrauchsfertigen Formulierung hervorbringt. produces ready-to-use formulation.
Ein weiterer Gegenstand der Erfindung ist ein Verfahren wie oben beschrieben, wobei das Insulin, das Insulinanalogon oder das Insulinderivat als kristalliner oder amorpher Feststoff vorliegt. Ein Gegenstand der Erfindung ist ein Verfahren wie oben beschrieben, wobei dasAnother object of the invention is a method as described above, wherein the insulin, the insulin analogue or the insulin derivative is present as a crystalline or amorphous solid. An object of the invention is a method as described above, wherein the
Insulin ausgewählt wird aus einer Gruppe enthaltend menschliches Insulin, Schweine- Insulin und Insulin des Rindes. Ein weiterer Gegenstand der Erfindung ist ein Verfahren wie oben beschrieben, wobei das Insulinanalogon ausgewählt wird aus einer Gruppe enthaltend Gly(A21), Arg(B31), Arg(B32)-Humaninsulin, Lys(B3), Glu(B29)-Humaninsulin, Asp(B28)-Humaninsulin, Lys(B28) Pro(B29)-Humaninsulin und Des(B30)-Humaninsulin. Insulin is selected from a group containing human insulin, porcine insulin and bovine insulin. The invention further provides a method as described above, wherein the insulin analog is selected from a group comprising Gly (A21), Arg (B31), Arg (B32) -human insulin, Lys (B3), Glu (B29) -human insulin, Asp (B28) human insulin, Lys (B28) Pro (B29) human insulin and Des (B30) human insulin.
Ein weiterer Gegenstand der Erfindung ist ein Verfahren gemäß einem oder mehreren der Ansprüche 1 bis 3, wobei das Insulinanalogon ausgewählt wird aus einer Gruppe enthaltend ein Insulinanalogon der Formel I Another object of the invention is a method according to one or more of claims 1 to 3, wherein the insulin analogue is selected from a group containing an insulin analog of the formula I.
S S S S
1 5 | 10| 15 20  1 5 | 10 | 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
I (SEQ ID NO: 1) \ A-Kette I (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; B30 Thr oder einer chemischen Bindung; B4 Asp, Glu or GIn; B29 Lys or a chemical bond; 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, vorzugsweise, bei der das Insulinanalogon ausgewählt ist aus einer Gruppe enthaltend: or a pharmacologically tolerable salt thereof, preferably 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 Humaninsulin, Arg (AO), His (A8), GIu (A5), Asp (A18), GIy (A21), Arg (B31), Lys (B32)- NH2 Humaninsulin, 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), 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 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)- NH2 Humaninsulin, Arg (AO), His (A8), GIu (A15), GIy (A21), Asp (B3), Arg (B31), Lys (B32)- NH2 Humaninsulin, Arg (AO), 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 Humaninsulin, 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), 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 Humaninsulin, 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 Humaninsulin, 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 Humaninsulin, Arg (AO), His (A8), Asp (A18), GIy (A21), Asp (B1), Arg (B31), Arg (B32)- NH2 Humaninsulin, Arg (AO), His (A8), Asp (A18), GIy (A21), Asp (B1), Arg (B31), Lys (B32)- NH2 Humaninsulin, 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 Humaninsulin, 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), His (A8), Glu (A5), Glu (A15), Gly (A21), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), 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 (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), His (A8), Asp (A18), Gly (A21), Asp (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 (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 (AO), 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), Glu (BO), Arg (B31), Arg (B3 2) - 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), Glu (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) , GIu (A15), Gly (A21), Asp (B1), Arg (B31), Lys (B32) - NH 2 human insulin, Arg (AO), His (A8), Asp (A18), Gly (A21), Asp (B1), Arg (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 human insulin, Arg (AO), His (A8), Asp (A18), Gly (A21), Asp (B3), Arg (B30), Lys (B31) - NH 2 human insulin.
Ein weiterer Gegenstand der Erfindung ist ein Verfahren wie oben beschrieben, wobei das Insulinanalogon ausgewählt wird aus einer Gruppe enthaltend Another object of the invention is a method as described above, wherein the insulin analogue is selected from a group containing
ein Insulinanalogon der Formel Il S S an insulin analog of formula II SS
5 I 10 I 15 20  5 I 10 I 15 20
A-I AO Al I V E A5 C C H S I C S L Y A15 L E Al 8 Y C A21  A-I AO Al I V E A5 C C H S I C S L Y A15 L E Al 8 Y C A21
A-Kette A chain
I I  I i
B-I BO Bl V 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 Bl V 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: 4) B-Kette T P B29 B30 B31 B32 (SEQ ID NO: 4) B chain
30 wobei  30 where
A-1 Lys, Arg oder einer Aminogruppe; A-1 Lys, Arg or an amino group;
AO Lys, Arg oder einer chemischen Bindung; AO Lys, Arg or a chemical bond;
A1 Arg oder GIy; A5 Asp, GIu oder GIn; A1 Arg or Gly; A5 Asp, Glu or GIn;
A15 Asp, GIu oder GIn; A15 Asp, Glu or GIn;
A18 Asp, GIu oder Asn; A18 Asp, Glu or Asn;
A21 AIa, Ser, Thr oder GIy; A21 AIa, Ser, Thr or GIy;
B-1 Asp, GIu oder eine Aminogruppe; BO Asp, GIu oder eine chemische Bindung; B-1 Asp, Glu or an amino group; BO Asp, Glu or a chemical bond;
B1 Asp, GIu, Phe oder eine chemische Bindung; B3 Asp, GIu oder Asn; B1 Asp, Glu, Phe or a chemical bond; B3 Asp, Glu or Asn;
B4 Asp, GIu oder GIn; B4 Asp, Glu or GIn;
B29 Arg, Lys oder einer Aminosäure ausgewählt aus einer Gruppe enthaltend die Aminosäuren Phe, AIa, Thr, Ser, VaI, Leu, GIu oder Asp, oder einer chemischen Bindung; B29 Arg, Lys or an amino acid selected from a group containing the amino acids Phe, Ala, Thr, Ser, VaI, Leu, Glu or Asp, or a chemical bond;
B30 Thr oder einer chemischen Bindung; B31 Arg, Lys oder einer chemischen Bindung; B32 Arg-Amid oder Lys-Amid entspricht, wobei nicht mehr als ein Aminosäurerest der Gruppe enthaltend A5, A15, A18, B-1 , BO, B1 , B2, B3 und B4 gleichzeitig und unabhängig voneinander Asp oder GIu entsprechen, vorzugsweise bei der das Insulinanalogon ausgewählt ist aus einer Gruppe enthaltend: B30 Thr or a chemical bond; B31 Arg, Lys or a chemical bond; B32 is Arg-amide or Lys-amide, wherein no more than one amino acid residue of the group containing A5, A15, A18, B-1, BO, B1, B2, B3 and B4 simultaneously and independently of one another correspond to Asp or Glu, preferably in the the insulin analog is selected from a group comprising:
Arg (A-1 ), Arg (AO), GIu (A5), His (A8), GIy (A21 ), Arg (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), GIu (A5), His (A8), GIy (A21 ), Lys (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), GIu (A15), His (A8), GIy (A21 ), Arg (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), GIu (A15), His (A8), GIy (A21 ), Lys (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), Asp (A18), His (A8), GIy (A21 ), Arg (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), Asp (A18), His (A8), GIy (A21 ), Arg (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), His (A8), GIy (A21 ), GIu (BO), Arg (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), His (A8), GIy (A21 ), GIu (BO), Lys (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), His (A8), GIy (A21 ), Asp (B3), Arg (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), His (A8), GIy (A21 ), Asp (B3), Lys (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), His (A8), GIy (A21 ), GIu (B4), Arg (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), His (A8), GIy (A21 ), GIu (B4), Lys (B30) - NH2 Humaninsulin, Arg (AO), His (A8), GIy (A21 ), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin, Arg (A-1), Arg (AO), Glu (A5), His (A8), Gly (A21), Arg (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), Glu ( A5), His (A8), Gly (A21), Lys (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), Glu (A15), His (A8), Gly (A21), Arg (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), Glu (A15), His (A8), Gly (A21), Lys (B30) - NH 2 human insulin, Arg (A-1) , Arg (AO), Asp (A18), His (A8), Gly (A21), Arg (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), Asp (A18), His (A8 ), Gly (A21), Arg (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), His (A8), Gly (A21), Glu (BO), Arg (B30) - NH 2 Human insulin, Arg (A-1), Arg (AO), His (A8), Gly (A21), Glu (BO), Lys (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), His (A8), Gly (A21), Asp (B3), Arg (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), His (A8), Gly (A21), Asp (B3) , Lys (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), His (A8), Gly (A21), Glu (B4), Arg (B30) - NH 2 human insulin, Arg (A-) 1), Arg (AO), His (A8), Gly (A21), Glu (B4), Lys (B3 0) - NH 2 human insulin, Arg (AO), His (A8), Gly (A21), Arg (B31), Arg (B32) - NH 2 - human insulin,
Arg (AO), His (A8), GIy (A21 ), Arg (B31 ), Lys (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), Gly (A21), Arg (B31), Lys (B32) - NH 2 - human insulin,
Arg (AO), GIu (A5), His (A8), GIy (A21 ), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin, Arg (AO), GIu (A5), His (A8), GIy (A21 ), Arg (B31 ), Lys (B32) - NH2 - Humaninsulin, Arg (AO), Asp (A18), His (A8), GIy (A21 ), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin, Arg (AO), Asp (A18), His (A8), GIy (A21 ), Arg (B31 ), Lys (B32) - NH2 - Humaninsulin, Arg (AO), GIu (A15), His (A8), GIy (A21 ), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin, Arg (AO), GIu (A15), His (A8), GIy (A21 ), Arg (B31 ), Lys (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), GIy (A21 ), Asp (B3), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), GIy (A21 ), Asp (B3), Arg (B31 ), Lys (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), GIy (A21 ), GIu (B4), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), GIy (A21 ), GIu (B4), Arg (B31 ), Lys (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), GIy (A21 ), GIu (BO), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), GIy (A21 ), GIu (BO), Arg (B31 ), Lys (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), GIy (A21 ), Arg (B30) - NH2 - Humaninsulin, Arg (AO), Glu (A5), His (A8), Gly (A21), Arg (B31), Arg (B32) - NH 2 - human insulin, Arg (AO), Glu (A5), His (A8), Gly (A21), Arg (B31), Lys (B32) - NH 2 - human insulin, Arg (AO), Asp (A18), His (A8), Gly (A21), Arg (B31), Arg (B32) - NH 2 - human insulin, Arg (AO), Asp (A18), His (A8), Gly (A21), Arg (B31), Lys (B32) - NH 2 - human insulin, Arg (AO), Glu (A15), His (A8), Gly (A21), Arg (B31), Arg (B32) - NH 2 - human insulin, Arg (AO), Glu (A15), His (A8), Gly (A21), Arg (B31), Lys (B32) - NH 2 - human insulin, Arg (AO), His (A8), Gly (A21), Asp (B3), Arg (B31), Arg (B32) - NH 2 - human insulin, Arg (AO), His (A8), Gly (A21), Asp (B3), Arg (B31), Lys (B32) - NH 2 - human insulin, Arg (AO), His (A8), Gly (A21), Glu (B4), Arg (B31), Arg (B32) - NH 2 - human insulin, Arg (AO), His (A8), Gly (A21), Glu (B4), Arg (B31), Lys (B32) - NH 2 - human insulin, Arg (AO), His (A8), Gly (A21), Glu (BO), Arg (B31), Arg (B32) - NH 2 - human insulin, Arg (AO), His (A8), Gly (A21), Glu (BO), Arg (B31), Lys (B32) - NH 2 - human insulin, Arg (AO), His (A8), Gly (A21), Arg (B3 0) - NH 2 - human insulin,
Arg (AO), His (A8), GIy (A21 ), Lys (B30) - NH2 - Humaninsulin, Arg (AO), His (A8), Gly (A21), Lys (B30) - NH 2 - human insulin,
Arg (A-1 ), Arg (AO), His (A8), GIy (A21 ), Arg (B30) - NH2 - Humaninsulin, Arg (A-1), Arg (AO), His (A8), Gly (A21), Arg (B30) - NH 2 - human insulin,
Arg (A-1 ), Arg (AO), His (A8), GIy (A21 ), Lys (B30) - NH2 - Humaninsulin, Arg (A-1), Arg (AO), His (A8), Gly (A21), Lys (B30) - NH 2 - human insulin,
Arg (AO), Arg (A1 ), His (A8), GIy (A21 ), Arg (B30) - NH2 - Humaninsulin, Arg (AO), Arg (A1), His (A8), Gly (A21), Arg (B30) - NH 2 - human insulin,
Arg (AO), Arg (A1 ), His (A8), GIy (A21 ), Lys (B30) - NH2 - Humaninsulin, Arg (AO), Arg (A1), His (A8), Gly (A21), Lys (B30) - NH 2 - human insulin,
His (A8), GIy (A21 ), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin. Ein weiterer Gegenstand der Erfindung ist ein Verfahren gemäß einem oder mehreren der Ansprüche 1 bis 3, wobei das Insulinderivat ausgewählt wird aus einer Gruppe enthaltend B29-N-myristoyl-des(B30) Humaninsulin, B29-N-palmitoyl-des(B30) Humaninsulin, B29-N-myhstoyl Humaninsulin, B29-N-palmitoyl Humaninsulin, B28-N- myristoyl LysB28ProB29 Humaninsulin, B28-N-palmitoyl-LysB28ProB29 Humaninsulin, B30-N-myhstoyl-ThrB29LysB3° Humaninsulin, B30-N-palmitoyl- ThrB29LysB30 His (A8), Gly (A21), Arg (B31), Arg (B32) - NH 2 - human insulin. A further subject of the invention is a method according to one or more of claims 1 to 3, wherein the insulin derivative is selected from a group containing B29-N-myristoyl des (B30) human insulin, B29-N-palmitoyl des (B30) human insulin , B29-N-myhstoyl human insulin, B29-N-palmitoyl human insulin, B28-N-myristoyl Lys B28 Pro B29 human insulin, B28-N-palmitoyl-Lys B28 Pro B29 human insulin, B30-N-myhstoyl-Thr B29 Lys B3 ° human insulin , B30-N-palmitoyl-Thr B29 Lys B30
Humaninsulin, B29-N-(N-palmitoyl-Y-glutamyl)-des(B39) Humaninsulin, B29-N-(N- lithocholyl-Y-glutamyl)-des(B30) Humaninsulin, B29-N-(ω-carboxyheptadecanoyl)- des(B30) Humaninsulin and B29-N-(ω-carboxyheptadecanoyl) Humaninsulin. Ein weiterer Gegenstand der Erfindung ist ein Verfahren wie oben beschrieben, wobei in der Formulierung ein Konservierungsmittel ausgewählt aus einer Gruppe enthaltend Phenol, m-Cresol, Chlorkresol, Benzylalkohol, Parabene vorhanden ist. Human insulin, B29-N- (N-palmitoyl-Y-glutamyl) -des (B39) human insulin, B29-N- (N-lithocholyl-Y-glutamyl) des (B30) human insulin, B29-N- (ω-carboxyheptadecanoyl ) - the (B30) human insulin and B29-N- (ω-carboxyheptadecanoyl) human insulin. Another object of the invention is a method as described above, wherein in the formulation a preservative selected from a group containing phenol, m-cresol, chlorocresol, benzyl alcohol, parabens is present.
Ein weiterer Gegenstand der Erfindung ist ein Verfahren wie oben beschrieben, wobei in der Formulierung ein Isotonisierungsmittel ausgewählt aus einer Gruppe enthaltend Mannitol, Sorbitol, Lactose, Dextrose, Trehalose, Natriumchlorid, Glycerol vorhanden ist. Another object of the invention is a method as described above, wherein in the formulation an isotonizing agent selected from a group containing mannitol, sorbitol, lactose, dextrose, trehalose, sodium chloride, glycerol is present.
Ein weiterer Gegenstand der Erfindung ist ein Verfahren wie oben beschrieben, wobei das Insulin, das Insulinanalogon und/oder das Insulinderivat in einer Konzentration von 240 - 3000 nmol/ml vorliegt Another object of the invention is a method as described above, wherein the insulin, the insulin analogue and / or the insulin derivative is present in a concentration of 240-3000 nmol / ml
Ein weiterer Gegenstand der Erfindung ist ein Verfahren wie oben beschrieben, wobei in der Formulierung 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, vorzugsweise bei dem ein Analogon von Exendin-4 ausgewählt wird aus einer Gruppe enthaltend Another object of the invention is a method as described above, wherein in the formulation additionally a glucagon-like peptide-1 (GLP1) or an analog or derivative thereof, or exendin-3 or -4 or an analog or derivative thereof is included , preferably in which an analog of exendin-4 is selected from a group comprising
H-desPro36-Exendin-4-Lys6-NH2, H-desPro 36 -Exendin-4-Lys 6 -NH 2 ,
H-des(Pro36 37)-Exendin-4-l_ys4-NH2 und H des (Pro 36 37 ) -Exendin-4-l_ys 4 -NH 2 and
H-des(Pro36 37)-Exendin-4-Lys5-NH2, H des (Pro 36 37 ) -Exendin-4-Lys 5 -NH 2 ,
oder ein pharmakologisch tolerierbares Salz davon. or a pharmacologically tolerable salt thereof.
Ein weiterer Gegenstand der Erfindung ist ein Verfahren wie oben beschrieben, bei dem ein Analogon von Exendin-4 ausgewählt wird aus einer Gruppe enthaltend desPro36 [Asp28] Exend in-4 (1 -39), Another object of the invention is a method as described above, wherein an analog of exendin-4 is selected from a group containing the Pro 36 [Asp 28 ] exend in-4 (1 -39),
desPro36 [IsoAsp28] Exend in-4 (1 -39), desPro 36 [IsoAsp 28 ] Exend in-4 (1 -39),
desPro36 [Met(O)14, Asp28] Exend in-4 (1 -39), DESpro 36 [Met (O) 14, Asp 28] exend in-4 (1 -39)
desPro36 [Met(O)14, IsoAsp28] Exend in-4 (1 -39), desPro 36 [Met (O) 14 , IsoAsp 28 ] Exend in-4 (1 -39),
desPro36 [Trp(O2)25, Asp28] Exend in-2 (1 -39), desPro 36 [Trp (O 2 ) 25 , Asp 28 ] Exend in-2 (1 -39),
desPro36 [Trp(O2)25, IsoAsp28] Exend in-2 (1 -39), desPro 36 [Trp (O 2 ) 25 , IsoAsp 28 ] Exend in-2 (1 -39),
desPro36 [Met(O)14Trp(O2)25, Asp28] Exend in-4 (1 -39) und desPro 36 [Met (O) 14 Trp (O 2 ) 25 , Asp 28 ] Exend in-4 (1 -39) and
desPro36 [Met(O)14Trp(O2)25, IsoAsp28] Exend in-4 (1-39), oder ein pharmakologisch tolerierbares Salz davon, vorzugsweise bei denen an die C- Termini der Analoga von Exendin-4 das Peptid -Lys6-NH2 angefügt ist. desPro 36 [Met (O) 14 Trp (O 2 ) 25 , IsoAsp 28 ] Exend in-4 (1-39), or a pharmacologically tolerable salt thereof, preferably wherein the peptide -Lys6-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) -Lys 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] Exend in-4(1 -39) -NH2, H-Asn- (Glu) 5 of the Pro 36 , Pro 37 , Pro 38 [Asp 28 ] Exend in-4 (1 -39) -NH 2 ,
des Pro36, Pro37, Pro38 [Asp28]Exendin-4(1 -39)-(Lys)6-NH2, Pro 36 , Pro 37 , Pro 38 [Asp 28 ] Exendin-4 (1 -39) - (Lys) 6 -NH 2 ,
H-(LyS)6- des Pro36, Pro37, Pro38 [Asp28]Exendin-4(1 -39)-(Lys)6-NH2, H- (LyS) 6 - of Pro 36 , Pro 37 , Pro 38 [Asp 28 ] Exendin-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] Exend in-4(1 -39) -NH2, H- the Asp 28 Pro 36, Pro 37, Pro 38 [Trp (O2) 25] exend in-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-(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, 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, Asp28]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 , 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-(Glu)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 the Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Asp 28 ] Exendin-4 (1 -39) -NH 2 , of the 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- (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 des Pro36, Pro37, Pro38 [Met(O)14, Asp28] Exendin-4(1 -39)-(Lys)6-NH2, H-(LyS)6- des Pro36 [Met(O)14, Trp(O2)25, Asp28]Exendin-4(1 -39)-Lys6-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 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, of Asp 28 Pro 36, Pro 37, Pro 38 [Met (O) 14, Trp (O2) 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] Exend in-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 ] Exend in-4 (1 -39) - (Lys) 6 - NH 2 .
H-Asn-(Glu)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 ein Verfahren wie oben beschrieben, wobei in der Formulierung zusätzlich Arg34, Lys26 (Nε(γ-glutamyl(Nα-hexadecanoyl))) GLP-1 (7-37) [liraglutide] oder ein pharmakologisch tolerierbares Salz davon enthalten ist. Another object of the invention is a method as described above, wherein in the formulation additionally Arg 34 , Lys 26 (N ε (γ-glutamyl (N α -hexadecanoyl))) GLP-1 (7-37) [liraglutide] or a pharmacologically tolerable salt thereof is included.
Ein weiterer Gegenstand der Erfindung ist ein Verfahren wie oben beschrieben, wobei in der Formulierung noch ein Zinksalz enthalten ist. Another object of the invention is a method as described above, wherein in the formulation nor a zinc salt is included.
Ein weiterer Gegenstand der Erfindung ist die Verwendung eines Verfahrens wie oben beschrieben bei der großtechnischen Herstellung eines Insulins, Insulinanalogs oder Insulinderivats. Another object of the invention is the use of a method as described above in the large-scale production of an insulin, insulin analog or insulin derivative.
Ein weiterer Gegenstand der Erfindung ist ein zweiteiliger Satz von Behältern, in denen einer der Behälter ein Insulin, ein Insulinanalogon oder ein Insulinderivat als Feststoff und der andere Behälter ein Lösemittelgemisch eines bestimmten pH-Werts mit der Endkonzentration der Exzipienten einer gewünschten Formulierung eines Insulins, eines Insulinanalogons oder eines Insulinderivats enthält; zur hitze- und schüttelstabilen Aufbewahrung des Insulins, des Insulinanalogons oder des Another object of the invention is a two-part set of containers, in which one of the containers an insulin, an insulin analogue or an insulin derivative as a solid and the other container a solvent mixture of a certain pH with the final concentration of the excipients of a desired formulation of an insulin, a Insulin analogues or an insulin derivative; for heat and shake-stable storage of insulin, the insulin analog or the
Insulinderivats für das spätere Herstellen einer gebrauchsfertigen Formulierung durch Lösen des Feststoffs in dem Lösemittelgemisch wie oben beschrieben. Insulin derivative for later preparing a ready-to-use formulation by dissolving the solid in the solvent mixture as described above.
Ein weiterer Gegenstand der Erfindung ist ein Zweikammerinjektionssystem, bei dem eine Kammer ein Insulin, ein Insulinanalogon oder ein Insulinderivat als Feststoff und die andere Kammer A further subject of the invention is a two-chamber injection system in which one chamber contains an insulin, an insulin analog or an insulin derivative as solid and the other chamber
ein Lösemittelgemisch eines bestimmten pH-Werts mit der Endkonzentration der Exzipienten einer gewünschten Formulierung eines Insulins, eines Insulinanalogons oder eines Insulinderivats enthält; zur hitze- und schüttelstabilen Aufbewahrung des Insulins, des Insulinanalogons oder des Insulindehvats für das spätere Herstellen einer gebrauchsfertigen Formulierung durch Lösen des Feststoffs in dem Lösemittelgemisch wie oben beschrieben. In dieser Beschreibung werden die Begriffe„Formulierung" und„Zubereitung" synonym verwendet. a solvent mixture of a certain pH containing the final concentration of the excipients of a desired formulation of an insulin, an insulin analog or an insulin derivative; for heat- and shake-stable storage of Insulin, the insulin analog or the insulin dilate for later preparing a ready-to-use formulation by dissolving the solid in the solvent mixture as described above. In this description, the terms "formulation" and "preparation" are used interchangeably.
Figurenlegende: Figure legend:
Fig. 1 : Blutzuckersenkende Wirkung von neuen Insulinanaloga gemäß Formel I in Ratten  Fig. 1: Hypoglycemic effect of new insulin analogues according to formula I in rats
Fig. 2: Blutzuckersenkende Wirkung von neuen Insulinanaloga gemäß Formel I im Fig. 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
Fig. 5: Blutzuckersenkende Wirkung von erfindungsgemäßen Insulinanaloga derFig. 5: Hypoglycemic effect of insulin analogues of the invention
Formel Il in Ratten Formula II in rats
Fig. 6: Blutzuckersenkende Wirkung von Insulin Glargin in Ratten  Fig. 6: Hypoglycemic effect of insulin glargine in rats
Beispiele: Examples:
Die folgenden Beispiele sollen den Erfindungsgedanken näher erläutern, ohne beschränkend zu wirken. Beispiel 1 : Vereinfachtes Auflösen von Insulinen in einem Schritt (Auflösetest) The following examples are intended to explain the concept of the invention in more detail without being restrictive. Example 1: Simplified dissolution of insulins in one step (dissolution test)
Üblicherweise werden Insuline in wäßrigen Systemen, die Hilfs-und Zusatzstoffe wie beispielsweise Antibakterizide, isotonisierende Agenzien, Puffersubstanzen, und/oder Tenside enthalten (= im folgenden„Lösemittel" genannt) auf dem Markt angeboten. Das Auflösen von Insulinen zur Herstellung pharmakologischer Zubereitung wird im Allgemeinen dadurch gewährleistet, daß die Insuline zunächst sauer angelöst werden, um dann in weiteren Schritten auf die erforderliche Konzentration und pH-Wert der Lösung eingestellt zu werden (The Wellcome Foundation Limited te London, Usually, insulins are available on the market in aqueous systems containing adjuvants and additives such as antibacterial agents, isotonizing agents, buffer substances, and / or surfactants (= "solvent") Generally ensures that the insulins are first dissolved sour, to then in further steps to the required concentration and pH of the Solution (The Wellcome Foundation Limited, London,
Octrooiraad Nederland, Octrooiannnvrage No. 6506714, 26.05.1965 "Werkwijze voor het bereiden van insulinepreparaten"). Alternativ ist ein Anlösen von Insulinen im Alkalischen einhergehend mit einer leicht erhöhten Stabilität der Insulinzubereitungen beschrieben (WO 2004/096266 Octrooiraad Nederland, Octrooiannnvrage No. 6506714, 26.05.1965 "Werkwijze voor het bereiden van insulinepreparaten"). Alternatively, a solubilization of insulins in the alkaline is described along with a slightly increased stability of insulin preparations (WO 2004/096266
PCT/DK2004/000300, "Improved Physical Stability of Insulin Formulations"). PCT / DK2004 / 000300, "Improved Physical Stability of Insulin Formulations").
Auflösetest: Dissolution test:
Um der komplizierten Prozedur des Auflösens von Insulinen zu entgehen (siehe oben), wurde getestet, ob sich Insuline als Feststoffe so auflösen lassen, daß ihre auf dem Markt befindlichen Formulierung in einem Schritt erreicht wird. Ziel war es, die To avoid the complicated procedure of dissolving insulins (see above), it was tested whether insulins can be solubilized as solids so that their formulation on the market is achieved in one step. The goal was, the
Komposition der Marktformulierung beizubehalten, d.h. unter Berücksichtigung der Konzentration aller Hilfs-und Zusatzstoffe und des pH-Wertes. Composition of the market formulation, i. taking into account the concentration of all auxiliaries and additives and the pH.
Es wurden untersucht: It was investigated:
a. Lantus® (Hilfs-und Zusatzstoffe Zinkchlorid, m-Kresol, Glyzerin, pH 4) b. Insuman® (Hilfs-und Zusatzstoffe Nathumhydrogenphosphat, m-Kresol, Glyzerin, pH 7.3) a. Lantus ® (auxiliaries and additives zinc chloride, m-cresol, glycerin, pH 4) b. Insuman ® (auxiliaries and additives Nathumhydrogenphosphat, m-cresol, glycerin, pH 7.3)
Es wurden verschiedene Lösemittelgemische zubereitet, welche der Endkonzentration der Exzipienten für die beiden Insuline Lantus® und Insuman® entsprachen. Hierbei wurde der pH-Wert des Lösemittels variiert. a. Lantus® erforderte einen pH-Wert des Lösemittels von pH 2.85, um eine komplette Auflösung mit dem erforderlichen End-pH-Wert von pH 4 zu erreichen. Die There were prepared various solvent mixtures, which corresponded to the final concentration of the excipient for the two Lantus ® and Insuman ®. In this case, the pH of the solvent was varied. a. Lantus ® required a pH of the solvent of pH 2.85 to achieve complete dissolution with the required final pH of pH 4. The
Auflösegeschwindigkeit (Endvolumen 1 mL) lag unter 10min. b. Insuman® erforderte einen pH-Wert des Lösemittels von pH 7.6, um eine komplette Auflösung mit dem erforderlichen End-pH-Wert von pH 7.3 zu erreichen. Die Dissolution rate (final volume 1 mL) was less than 10 minutes. b. Insuman ® required a pH of 7.6 to reach a complete solution with the required final pH of 7.3. The
Auflösegeschwindigkeit (Endvolumen 1 mL) lag unter 10min. LC-UV/MS-Analyse der Proben im Anschluß an die vereinfachte Auflösung mit anschließender Auswertung des UV-Chromatograms ergab mit den Dissolution rate (final volume 1 mL) was less than 10 minutes. LC-UV / MS analysis of the samples following the simplified resolution followed by evaluation of the UV chromatogram revealed
Marktformulierungen identische Spektren (UV-Signal des m-Kresols und des jeweiligen Insulins hinsichtlich Retentionszeit und Peakfläche). Market formulations identical spectra (UV signal of the m-cresol and the respective insulin in terms of retention time and peak area).
Schlußfolgerung: Es ist möglich, Insuline in einem Schritt zu lösen, d.h. ohne vorheriges Anlösen im sauren oder alkalischen pH. Als Schlüsselfaktor wurde die Anpassung des pH-Werts des Lösemittels auf die jeweils zu erzielende Konzentration sowie den pH-Wert des fertigen Produktes in Abhängigkeit der intrinsischen, den pH- Wert verändernden Eigenschaften der zu lösenden Insuline identifiziert. Unter Conclusion: It is possible to solve insulins in one step, i. without previous dissolution in acidic or alkaline pH. As a key factor, the adaptation of the pH of the solvent to the particular concentration to be achieved and the pH of the finished product were identified as a function of the intrinsic, pH-changing properties of the insulins to be dissolved. Under
Anpassung dieser empirisch ermittelten Parameter konnte der Auflöseprozeß für die Insuline in einem angemessenen Zeitrahmen (<10 min) realisiert werden.  Adapting these empirically determined parameters, the dissolution process for the insulins could be realized within a reasonable time frame (<10 min).
Beispiel 2: Hitzestabilität von Feststoffen (Hitzestresstest, Schütteltest, Amorphe Feststoffe) Example 2: Heat Stability of Solids (Heat Stress Test, Shake Test, Amorphous Solids)
Die Hitzestabilität von wäßrigen Insulinzubereitungen ist Gegenstand zahlreicher Untersuchungen (Literatur: A. Krogh, A. M. Hemmingsen, Biochem. J. (1928) 22, 1231 -1238 "The destructive action of heat on insulin Solutions"). The heat stability of aqueous insulin preparations has been the subject of numerous studies (literature: A. Krogh, A.M. Hemmingsen, Biochem J. (1928) 22, 1231-1238 "The destructive action of heat on insulin solutions").
Es ist bekannt, daß Zusatzstoffe unterschiedlich zur thermischen Stabilität des gelösten Insulins beitragen, z.B. die Zinkkonzentration oder der pH-Wert (N. R. It is known that additives contribute differently to the thermal stability of the dissolved insulin, e.g. the zinc concentration or the pH (N.R.
Stephenson, R. G. Romans; Journal of Pharmacy and Pharmacology (1960), 12 372- 376 "Thermal Stability of Insulin made from Zinc Insulin Crystals"). Stephenson, R.G. Romans; Journal of Pharmacy and Pharmacology (1960), 12 372-376 "Thermal Stability of Insulin Made from Zinc Insulin Crystals").
Bei der Untersuchung von Hitzestabilität von hauseigenen, marktfertigen Lösungen, welche alle Hilfs-und Zusatzstoffe enthielten, konnte auch in eigenen Studien die Präzipitation - neben der Bildung von Degradationsprodukten - als Hauptursache für den Verlust von biologisch aktivem Insulinmaterial identifiziert werden (B.V. Fisher, P.B. Porter; J. Pharm. Pharmacol. (1981 ), 33 203-206 "Stability of bovine Insulin"). Aufgrund dieser Beobachtungen wurde die Hitzestabiliät des Insulinfeststoffes überprüft, wobei im folgenden die verwendeten Markennamen der fertigen, wäßrigen Formulierungen auch für die jeweiligen Insulinfeststoffe (Wirkstoffe) verwendet wurden: In the study of heat stability of in-house market solutions containing all excipients and additives, the precipitation - in addition to the formation of degradation products - could be identified as the main cause of loss of biologically active insulin material in own studies (BV Fisher, PB Porter J. Pharm. Pharmacol. (1981), 33 203-206 "Stability of bovine insulin"). On the basis of these observations, the heat stability of the insulin solid was checked, the brand names of the finished, aqueous formulations used being also used for the respective insulin solids (active substances) in the following:
Hitzestresstest: Heat stress test:
I) Verschiedene Insulinfeststoffe wurden bei unterschiedlichen Temperaturen (500C, 600C und 80°C) (Ausschluß von Licht) zwei Wochen lang gelagert. a. Lantus® (kristallin) ; b. Insuman® (kristallin); c. Apidra® (amporph) I) insulin Various solids were stored for two weeks at different temperatures (50 0 C, 60 0 C and 80 ° C) (absence of light). a. Lantus ® (crystalline); b. Insuman ® (crystalline); c. Apidra ® (amporph)
II) Verschiedene Insulinfeststoffe wurden bei 600C unterschiedlich lang II) Different insulin solids were at 60 0 C different lengths
gelagert (14 Tage, 1 Monat, 2 Monate, 3 Monate).  stored (14 days, 1 month, 2 months, 3 months).
a. Lantus® (kristallin) ; b. Insuman® (kristallin); c. Apidra® (amporph) a. Lantus ® (crystalline); b. Insuman ® (crystalline); c. Apidra ® (amporph)
III) Feststoffe von Arg (AO), His (A8), GIu (A15), Asp (A18), GIy (A21 ), Arg III) solids of Arg (AO), His (A8), Glu (A15), Asp (A18), Gly (A21), Arg
(B31 ), Arg (B32) Humaninsulinamid und Lantus®, beide in amorphem Zustand, wurden zwei Wochen lang bei 60°C gelagert. Nach Beendigung der Hitzestressphasen wurden die Feststoffe aufgelöst, so daß sich eine identische Konzentration wie die Marktformulierungen ergab. (B31), Arg (B32) Humaninsulinamid and Lantus ®, both in the amorphous state were stored at 60 ° C for two weeks. Upon completion of the heat stress phases, the solids were dissolved to give an identical concentration as the market formulations.
Als Vergleichsproben wurden Insulinlösungen mit gleicher Konzentration direkt vor der Messung gelöst. As comparative samples, insulin solutions with the same concentration were dissolved immediately before the measurement.
Zusätzlich wurden Lantus®, Insuman® und Apidra® in marktfertigen Zubereitungen/In addition, Lantus ® , Insuman ® and Apidra ® were marketed preparations /
Behältnissen (Quelle: Produktion) für 14 Tage bei 60°C gestreßt. Als Vergleichsproben wurden Behältnisse aus der gleichen Produktionscharge bei 4°C gelagert und zusammen mit den gestreßten Proben analysiert. Schlußfolgerung: Die Hitzestabilität von Insulinen bei Lagerung als Feststoff für 14 Tage erwies sich als temperaturabhängig. Der Grad der Stabilität war insulinabhängig. Die Lagerung als Feststoff erwies sich als hitzestabiler als die Lagerung in marktfertiger Lösung. Containers (source: production) for 14 days at 60 ° C. As comparison samples, containers from the same production batch were stored at 4 ° C. and analyzed together with the stressed samples. Conclusion: The heat stability of insulins when stored as a solid for 14 days proved to be temperature dependent. The degree of stability was insulin dependent. Storage as a solid proved to be more heat stable than storage in market ready solution.
(I) LC-UV/MS-Analyse der Proben mit anschließender Auswertung des UV-Chroma- tograms ergab folgende Werte: (I) LC-UV / MS analysis of the samples with subsequent evaluation of the UV chromatogram yielded the following values:
Schlußfolgerung: Die Hitzestabilität von Insulinen bei Lagerung als Feststoff für 14 Tage erwies sich als temperaturabhängig. Der Grad der Stabilität warinsulinabhängig. Die Lagerung als Feststoff erwies sich als hitzestabiler als die Lagerung in Conclusion: The heat stability of insulins when stored as a solid for 14 days proved to be temperature dependent. The degree of stability was insulin-dependent. Storage as a solid proved to be more heat stable than storage in
marktfertiger Lösung. (II) LC-UV/MS-Analyse der Proben mit anschließender Auswertung des UV-Chroma- tograms ergab folgende Werte:  Market ready solution. (II) LC-UV / MS analysis of the samples with subsequent evaluation of the UV chromatogram revealed the following values:
Lantus®: Lantus ®:
Insuman ® :. Insuman ®:.
Apidra®: Apidra ®:
Schlußfolgerung: Die Hitzestabilität von Insulinen bei Lagerung als Feststoff bei 600C erwies sich als zeitabhängig. Der Grad der Stabilität war insulinabhängig. (III) LC-UV/MS-Analyse der Proben mit anschließender Auswertung des UV-Chroma- tograms ergab folgende Werte: Conclusion: The heat stability of insulins when stored as a solid at 60 0 C proved to be time-dependent. The degree of stability was insulin dependent. (III) LC-UV / MS analysis of the samples with subsequent evaluation of the UV chromatogram gave the following values:
Schlussfolgerung: Die Hitzestabilität von Insulinen bei Lagerung als Feststoff bei 60°C erwies sich als zeitabhängig. Der Grad der Stabilität war insulinabhängig. Beide untersuchten langwirksamen Insuline erwiesen sich auch in amorphem Zustand als relativ hitzestabil. Conclusion: The heat stability of insulins when stored as solid at 60 ° C proved to be time-dependent. The degree of stability was insulin dependent. Both Long-acting insulins were found to be relatively heat-stable even in an amorphous state.
Schütteltest: shaking test:
Es ist bekannt, daß wäßrige Insulinzubereitungen gegenüber Schütteln instabil sind (A. Oliva, J.B. Farina, M. Llabres; Int. J. Of Pharmaceutics (1996), 143 143-170 It is known that aqueous insulin preparations are unstable to shaking (A. Oliva, J.B. Farina, M. Llabres, Int J. Of Pharmaceutics (1996) 143 143-170
"Influence of temperature and shaking on stability of insulin preparations: degradation kinetics"). Verschiedene Insulinformulierungen wurden bei Raumtemperatur (Ausschluß von Licht) 100h lang bei 1400rpm geschüttelt. a) Lantus®, Insuman® und Apidra® in marktfertigen Zubereitungen/Behältnissen (Quelle: Produktion) "Influence of temperature and shaking on the stability of insulin preparations: degradation kinetics"). Various insulin formulations were shaken at room temperature (exclusion of light) for 100 h at 1400 rpm. a) Lantus ®, Insuman ® and Apidra ® into marketable finished preparations / containers (Source: Production)
Lantus (Kartusche ohne Polysorbat 20, vial ohne Polysorbat 20, vial mit Lantus (cartridge without polysorbate 20, vial without polysorbate 20, vial with
Polysorbat 20), Insuman® (Kartusche, vial), Apidra® (Kartusche, vial) b) Lantus®, Insuman® und Apidra® als Feststoff (separat geschüttelt: Lösemittel zum direkten Auflösen in einem Schritt, siehe„Direktes Auflösen von Polysorbate 20), Insuman ® (cartridge, vial), Apidra ® (cartridge, vial) b) Lantus ® , Insuman ® and Apidra ® as solid (shaken separately: solvent for direct dissolution in one step, see "Direct dissolution of
Insulinfeststoff").  Insulin solid ").
Nach Beendigung der Schüttelphase wurden die Feststoffe (Proben b) aufgelöst, so daß sich eine identische Konzentration wie die Marktformulierungen ergab. c) Als Vergleichsproben wurden die entsprechenden Marktformulierungen auch 100h bei Raumtemperatur unter Ausschluß von Licht gelagert. After completion of the shaking phase, the solids (samples b) were dissolved to give an identical concentration as the market formulations. c) As comparative samples, the corresponding market formulations were also stored for 100 h at room temperature in the absence of light.
LC-UV/MS-Analyse der Proben mit anschließender Auswertung des UV-Chroma- tograms ergab folgende Werte (Referenz: Proben c): LC-UV / MS analysis of the samples followed by evaluation of the UV chromatogram yielded the following values (reference: samples c):
Schlußfolgerung: Mechanischer Streß durch Schütteln bewirkte auf Insulinlösungen eine Prezipitation von Insulin. Das Ausmaß ist insulinabhängig: Insuman® zeigte hierbei die größte Instabilität. Conclusion: Mechanical stress caused by shaking caused insulin solutions to precipitate insulin. The extent is insulin-dependent: Insuman ® showed the greatest instability.
Mechanischer Streß zeigte auf Insulin-Feststoff keine negative Auswirkungen: der Feststoff ließ sich anschließend zu 100% auflösen, auch wenn die Lösungen separat mitgeschüttelt wurden. Beispiel 3: Mögliche Anwendungen (großtechnische Herstellung von Mechanical stress did not show any negative effects on insulin solid: the solid could then be 100% dissolved, even if the solutions were shaken separately. Example 3: Possible applications (large-scale production of
Insulinzubereitungen, hitze- und schüttelstabile Darreichungsform in einem Zwei- Komponenten-System) Großtechnische Herstellung von Insulinzubereitungen: Insulin preparations, heat- and shake-stable dosage form in a two-component system) Large-scale production of insulin preparations:
Bei einer Auflösung in einem Schritt handelt es sich um ein vereinfachtes Verfahren mit folgenden Vorteilen: Es sind weniger Schritte mit weniger Zwischenanalysen (z.B. pH-Wert) zu erstellen, d.h. die Herstellung der Insulinzubereitungen kann schneller erfolgen. Außerdem ist die Herstellung weniger kompliziert, was eine vereinfachte Einarbeitung der Mitarbeiter mit sich bringt (weniger komplizierte SOPs). Zudem sind weniger Behälter zu reinigen, was wiederum Arbeitszeit und -material spart.  Single-step resolution is a simplified process with the following advantages: fewer steps with fewer intermediate analyzes (e.g., pH) must be made, i. The production of insulin preparations can be done faster. In addition, the manufacture is less complicated, resulting in a simplified training of the employees (less complicated SOPs). In addition, fewer containers are to be cleaned, which in turn saves working time and material.
Hitze- und Schüttelstabile Darreichungsform in einem Zwei-Komponenten-System: Es zeigte sich, daß feste Insuline - auch über einen längeren Zeitraum - bei Heat- and Shake-Stable Dosage Form in a Two-Component System: It has been shown that solid insulins - even over a longer period of time - are included
Hitzestress stabiler sind als gelöste. Es fand in geringem Ausmaß Degradationen statt, ein Verlust durch Präzipitation entfiel jedoch, so daß in der Folge bei vergleichbarem Hitzestress mehr bioverfügbares Insulin zur Verfügung stand, wenn der Lösevorgang erst nach dem Hitzestress stattfand. Präzipitation und Degradation, induziert durch starkes Schütteln der festen Insuline, konnte nicht beobachtet werden.  Heat stress are more stable than dissolved. Degradation took place to a small extent, but loss by precipitation was eliminated, so that more bioavailable insulin was available with comparable heat stress if the dissolution process took place after the heat stress. Precipitation and degradation induced by vigorous shaking of the solid insulins could not be observed.
Eine Darreichung in fester Form kann somit als generell temperaturstabiler sowie robuster gegenüber Schütteln und in Folge dessen als sicherer beschrieben werden. Darüberhinaus bietet eine Darreichung der Insuline in fester Form (Pulver) den zusätzlichen Vorteil, daß ein versehentliches Einfrieren durch den Patienten zu keiner Komplikation führen kann; feste Insuline werden bekanntermaßen in gefrorenem Zustand gelagert.  A presentation in solid form can thus be described as generally more temperature-stable and more robust to shaking and consequently safer. Moreover, administration of the insulins in solid form (powder) offers the additional advantage that accidental freezing by the patient can not lead to complication; Solid insulins are known to be stored in the frozen state.
Mögliche Anwendung: Insulinpulver statt gelöste Insuline werden mit geeignetem Lösemittel (im allgemeinen wäßrige Systeme, die Hilfs-und Zusatzstoffe wie  Possible use: Insulin powders instead of dissolved insulins are treated with suitable solvents (generally aqueous systems containing auxiliary agents and additives such as
beispielsweise Antibakterizide, isotonisierende Agenzien, Puffersubstanzen, und/oder Tenside enthalten = im folgenden„Lösemittel" genannt) in einem Zwei-Komponenten- System angeboten. Ein Zwei-Kammer-System ist beispielsweise beschrieben in WO2007/038773 A1. Dadurch erhöht sich der tolerierbare Temperaturbereich (sowohl in Richtung tieferer als auch höherer Temperaturen). Außerdem erhöht sich die Stabilität gegenüber mechanischem Streß wie zum Beispiel starke Erschütterungen. In der Folge sind längere Haltbarkeitsdauern, geringere Kosten für Lagerung und Transport und eine sicherere Arzneimittelanwendung zu erwarten. For example, antibacterial agents, isotonizing agents, buffering agents, and / or surfactants = hereinafter referred to as "solvents") are offered in a two-component system A two-chamber system is described, for example, in WO2007 / 038773 A1. This increases the tolerable temperature range (both towards lower and higher temperatures). In addition, the stability to mechanical stress such as strong vibrations increases. As a result, longer shelf lives, lower storage and transportation costs, and safer drug use are expected.
Durch das vereinfachte Auflösen in einem Schritt ist die Anwendbarkeit durch den Patienten gewährleistet.  Simplified dissolution in one step ensures applicability by the patient.
Beispiel 4: Formulierung der amidierten Indulinderivate Example 4: Formulation of the amidated induline 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. Als Lösungsmedium wurden folgende Zusammensetzungen verwendet: 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). 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 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 Concentration needed weighed. Thereafter, the present concentration was determined by analytical HPLC and the solution was then added to the 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 of the invention, no optimization of the formulations, e.g. with regard to an addition of isotonic agents, preservatives or buffer substances.
Beispiel 5: 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. 1 ), dass das eingesetzte erfindungsgemäße Insulinanalogon zu einem deutlich verzögerten Example 5: Evaluation of the hypoglycemic effect of novel insulin analogues in the rat The hypoglycemic effect of selected new insulin analogues 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 (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 prior to injection of the insulin analog and at regular intervals up to forty-eight hours after injection, blood samples are taken from the animals and it determines the blood sugar content. 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 twice the dose
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. 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. Beispiel 9: Formulierung der amidierten Insulinderivate The experiments were carried out as described in Example 35. FIG. 4 shows the result. Thereafter, the time-response 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: 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 of the invention, no optimization of the formulations, e.g. with regard to an addition of isotonic agents, preservatives or buffer substances.
Beispiel 10: Evaluierung der blutzuckersenkenden Wirkung von neuen Insulinanaloga in der Ratte Example 10: Evaluation of the hypoglycemic effect of novel 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. 5), dass das erfindungsgemäße Insulinanalogon zu einem deutlich verzögerten Wirkeintritt und einer längeren, gleichmäßigen Wirkdauer führt. 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. 5) 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 Example 11: Evaluation of the hypoglycemic 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, dass das erfindungsgemäße Insulinanalogon zu einem deutlich verzögerten, flachen Wirkeintritt und einer längeren, gleichmäßigen Wirkdauer führt. 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. Verfahren zur Herstellung einer wässrigen, pharmazeutischen Formulierung enthaltend ein Insulin, ein Insulinanalogon oder ein Insulinderivat, oder ein A process for the preparation of an aqueous pharmaceutical formulation comprising an insulin, an insulin analog or an insulin derivative, or a
pharmakologisch tolerierbares Salz davon, wobei die gebrauchsfertige Formulierung unmittelbar durch Lösen des Insulins, des Insulinanalogons oder des Insulinderivats als Feststoff mit einem geeigneten Lösemittelgemisch erfolgt. pharmacologically tolerable salt thereof, wherein the ready-to-use formulation is effected directly by dissolving the insulin, the insulin analogue or the insulin derivative as a solid with a suitable solvent mixture.
2. Verfahren gemäß Anspruch 1 , bei dem die Zusammensetzung des geeigneten Lösem ittelgemischs ermittelt wird, indem 2. The method according to claim 1, wherein the composition of the appropriate Lösel ittelgemischs is determined by
(a) Lösemittelgemische unterschiedlichen pH-Werts mit Konzentrationen von  (a) solvent mixtures of different pH with concentrations of
Exzipienten zubereitet werden, welche der Endkonzentration der Exzipienten der Formulierung enthaltend ein Insulin, ein Insulinanalogon oder ein Insulinderivat entsprechen und Excipients are prepared which correspond to the final concentration of the excipients of the formulation containing an insulin, an insulin analog or an insulin derivative, and
(b) durch Lösen des gewünschten Insulins, Insulinanalogons oder Insulinderivats dasjenige Lösemittelgemisch ermittelt wird, welches nach Lösen des Feststoffes von Insulin, Insulinanalogon oder Insulinderivat den gewünschten pH-Wert der (B) by dissolving the desired insulin, insulin analogue or insulin derivative that solvent mixture is determined which, after dissolving the solid of insulin, insulin analogue or insulin derivative, the desired pH of the
gebrauchsfertigen Formulierung hervorbringt. produces ready-to-use formulation.
3. Verfahren gemäß einem oder mehreren der Ansprüche 1 oder 2, wobei das Insulin, das Insulinanalogon oder das Insulinderivat als kristalliner oder amorpher Feststoff vorliegt. 3. The method according to one or more of claims 1 or 2, wherein the insulin, the insulin analogue or the insulin derivative is present as a crystalline or amorphous solid.
4. Verfahren gemäß einem oder mehreren der Ansprüche 1 bis 3, wobei das Insulin ausgewählt wird aus einer Gruppe enthaltend menschliches Insulin, Schweine-Insulin und Insulin des Rindes. 4. The method according to one or more of claims 1 to 3, wherein the insulin is selected from a group containing human insulin, porcine insulin and bovine insulin.
5. Verfahren gemäß einem oder mehreren der Ansprüche 1 bis 3, wobei das 5. The method according to one or more of claims 1 to 3, wherein the
Insulinanalogon ausgewählt wird aus einer Gruppe enthaltend Gly(A21 ), Arg(B31 ), Arg(B32)-Humaninsulin, Lys(B3), Glu(B29)-Humaninsulin, Asp(B28)-Humaninsulin, Lys(B28) Pro(B29)-Humaninsulin und Des(B30)-Humaninsulin. Insulin analog is selected from a group containing Gly (A21), Arg (B31), Arg (B32) human insulin, Lys (B3), Glu (B29) human insulin, Asp (B28) human insulin, Lys (B28) Pro (B29 ) Human insulin and Des (B30) human insulin.
6. Verfahren gemäß einem oder mehreren der Ansprüche 1 bis 3, wobei das Insulinanalogon ausgewählt wird aus einer Gruppe enthaltend 6. The method according to one or more of claims 1 to 3, wherein the insulin analogue is selected from a group containing
ein Insulinanalogon der Formel I  an insulin analog of the formula I.
s s
1 5 | 10| 15 20  1 5 | 10 | 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
I (SEQ ID NO: 1) \ A-Kette I (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  30
wobei in which
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; B1 Asp, Glu or Phe;
B2 Asp, GIu oder VaI; B3 Asp, GIu oder Asn; B2 Asp, Glu or VaI; B3 Asp, Glu or Asn;
B4 Asp, GIu oder GIn; B4 Asp, Glu or GIn;
B29 Lys oder einer chemischen Bindung; B29 Lys or a chemical bond;
B30 Thr oder einer chemischen Bindung; B31 Arg, Lys oder einer chemischen Bindung; 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 B30 Thr or a chemical bond; B31 Arg, Lys or a chemical bond; 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. or a pharmacologically tolerable salt thereof.
7. Verfahren gemäß Anspruch 6, bei der das Insulinanalogon ausgewählt ist aus einer Gruppe enthaltend: 7. The method of claim 6, 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 Humaninsulin, Arg (AO), His (A8), GIu (A5), Asp (A18), GIy (A21), Arg (B31), Lys (B32)- NH2 Humaninsulin, 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), 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 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)- NH2 Humaninsulin, Arg (AO), His (A8), GIu (A15), GIy (A21), Asp (B3), Arg (B31), Lys (B32)- NH2 Humaninsulin, Arg (AO), 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 Humaninsulin, 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), 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 Humaninsulin, 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 Humaninsulin, 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 Humaninsulin, Arg (AO), His (A8), Asp (A18), GIy (A21), Asp (B1), Arg (B31), Arg (B32)- NH2 Humaninsulin, Arg (AO), His (A8), Asp (A18), GIy (A21), Asp (B1), Arg (B31), Lys (B32)- NH2 Humaninsulin, 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 Humaninsulin, 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), His (A8), Glu (A5), Glu (A15), Gly (A21), Arg (B31), Arg (B32) - NH 2 human insulin, Arg (AO), 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 (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), His (A8), Asp (A18), Gly (A21), Asp (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 (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 (AO), 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), Glu (BO), Arg (A21) 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), Glu (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 (A) 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 (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 human insulin, Arg (AO), His (A8), Asp (A18), Gly (A21), Asp (B3), Arg (B30), Lys (B31) - NH 2 human insulin.
8. Verfahren gemäß einem oder mehreren der Ansprüche 1 bis 3, wobei das 8. The method according to one or more of claims 1 to 3, wherein the
Insulinanalogon ausgewählt wird aus einer Gruppe enthaltend Insulin analog is selected from a group containing
ein Insulinanalogon der Formel Il S S an insulin analog of formula II SS
1 5 | 10 I 15 20 A-I AO Al I V E A5 C C H S I C S L Y A15 L E A18 Y C A21  1 5 | 10 I 15 20 A-I AO Al I V E A5 C C H S I C S L Y A15 L E A18 Y C A21
A-Kette A chain
I I  I i
B-I BO Bl V 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 Bl V 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 T P B29 B30 B31 B32 (SEQ ID NO 4) B-Kette  SEQ ID NO 4) B chain
30 V V - ;  30 V V -;
wobei in which
A-1 Lys, Arg oder einer Aminogruppe;  A-1 Lys, Arg or an amino group;
AO Lys, Arg oder einer chemischen Bindung;  AO Lys, Arg or a chemical bond;
A1 Arg oder GIy; A1 Arg or Gly;
A5 Asp, GIu oder GIn;  A5 Asp, Glu or GIn;
A15 Asp, GIu oder GIn;  A15 Asp, Glu or GIn;
A18 Asp, GIu oder Asn;  A18 Asp, Glu or Asn;
A21 AIa, Ser, Thr oder GIy;  A21 AIa, Ser, Thr or GIy;
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, Phe oder eine chemische Bindung; B3 Asp, GIu oder Asn; B4 Asp, GIu oder GIn; BO Asp, Glu or a chemical bond; B1 Asp, Glu, Phe or a chemical bond; B3 Asp, Glu or Asn; B4 Asp, Glu or GIn;
B29 Arg, Lys oder einer Aminosäure ausgewählt aus einer Gruppe enthaltend die Aminosäuren Phe, AIa, Thr, Ser, VaI, Leu, GIu oder Asp, oder einer chemischen Bindung; B29 Arg, Lys or an amino acid selected from a group containing the amino acids Phe, Ala, Thr, Ser, VaI, Leu, Glu or Asp, or a chemical bond;
B30 Thr oder einer chemischen Bindung; B30 Thr or a chemical bond;
B31 Arg, Lys oder einer chemischen Bindung; B32 Arg-Amid oder Lys-Amid entspricht, wobei nicht mehr als ein Aminosäurerest der Gruppe enthaltend A5, A15, A18, B-1 , BO, B1 , B2, B3 und B4 gleichzeitig und unabhängig voneinander Asp oder GIu entsprechen. B31 Arg, Lys or a chemical bond; B32 is Arg amide or Lys amide, wherein no more than one amino acid residue of the group containing A5, A15, A18, B-1, BO, B1, B2, B3 and B4 simultaneously and independently corresponds to Asp or Glu.
9. Verfahren gemäß Anspruch 6, bei der das Insulinanalogon ausgewählt ist aus einer Gruppe enthaltend: 9. The method of claim 6, wherein the insulin analog is selected from a group comprising:
Arg (A-1 ), Arg (AO), GIu (A5), His (A8), GIy (A21 ), Arg (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), GIu (A5), His (A8), GIy (A21 ), Lys (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), GIu (A15), His (A8), GIy (A21 ), Arg (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), GIu (A15), His (A8), GIy (A21 ), Lys (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), Asp (A18), His (A8), GIy (A21 ), Arg (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), Asp (A18), His (A8), GIy (A21 ), Arg (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), His (A8), GIy (A21 ), GIu (BO), Arg (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), His (A8), GIy (A21 ), GIu (BO), Lys (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), His (A8), GIy (A21 ), Asp (B3), Arg (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), His (A8), GIy (A21 ), Asp (B3), Lys (B30) - NH2 Humaninsulin, Arg (A- 1 ), Arg (AO), His (A8), GIy (A21 ), GIu (B4), Arg (B30) - NH2 Humaninsulin, Arg (A-1 ), Arg (AO), His (A8), GIy (A21 ), GIu (B4), Lys (B30) - NH2 Humaninsulin, Arg (AO), His (A8), GIy (A21 ), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin, Arg (A-1), Arg (AO), Glu (A5), His (A8), Gly (A21), Arg (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), Glu ( A5), His (A8), Gly (A21), Lys (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), Glu (A15), His (A8), Gly (A21), Arg (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), Glu (A15), His (A8), Gly (A21), Lys (B30) - NH 2 human insulin, Arg (A-1) , Arg (AO), Asp (A18), His (A8), Gly (A21), Arg (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), Asp (A18), His (A8 ), Gly (A21), Arg (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), His (A8), Gly (A21), Glu (BO), Arg (B30) - NH 2 Human insulin, Arg (A-1), Arg (AO), His (A8), Gly (A21), Glu (BO), Lys (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), His (A8), Gly (A21), Asp (B3), Arg (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), His (A8), Gly (A21), Asp (B3) , Lys (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), His (A8), Gly (A21), Glu (B4), Arg (B30) - NH 2 human insulin, Arg (A-1), Arg (AO), His ( A8), Gly (A21), Glu (B4), Lys (B30) - NH 2 human insulin, Arg (AO), His (A8), Gly (A21), Arg (B31), Arg (B32) - NH 2 - human insulin,
Arg (AO), His (A8), GIy (A21 ), Arg (B31 ), Lys (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), Gly (A21), Arg (B31), Lys (B32) - NH 2 - human insulin,
Arg (AO), GIu (A5), His (A8), GIy (A21 ), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin, Arg (AO), GIu (A5), His (A8), GIy (A21 ), Arg (B31 ), Lys (B32) - NH2 - Humaninsulin, Arg (AO), Asp (A18), His (A8), GIy (A21 ), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin, Arg (AO), Asp (A18), His (A8), GIy (A21 ), Arg (B31 ), Lys (B32) - NH2 - Humaninsulin, Arg (AO), GIu (A15), His (A8), GIy (A21 ), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin, Arg (AO), GIu (A15), His (A8), GIy (A21 ), Arg (B31 ), Lys (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), GIy (A21 ), Asp (B3), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), GIy (A21 ), Asp (B3), Arg (B31 ), Lys (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), GIy (A21 ), GIu (B4), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), GIy (A21 ), GIu (B4), Arg (B31 ), Lys (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), GIy (A21 ), GIu (BO), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), GIy (A21 ), GIu (BO), Arg (B31 ), Lys (B32) - NH2 - Humaninsulin, Arg (AO), His (A8), GIy (A21 ), Arg (B30) - NH2 - Humaninsulin, Arg (AO), Glu (A5), His (A8), Gly (A21), Arg (B31), Arg (B32) - NH 2 - human insulin, Arg (AO), Glu (A5), His (A8), Gly (A21), Arg (B31), Lys (B32) - NH 2 - human insulin, Arg (AO), Asp (A18), His (A8), Gly (A21), Arg (B31), Arg (B32) - NH 2 - human insulin, Arg (AO), Asp (A18), His (A8), Gly (A21), Arg (B31), Lys (B32) - NH 2 - human insulin, Arg (AO), Glu (A15), His (A8), Gly (A21), Arg (B31), Arg (B32) - NH 2 - human insulin, Arg (AO), Glu (A15), His (A8), Gly (A21), Arg (B31), Lys (B32) - NH 2 - human insulin, Arg (AO), His (A8), Gly (A21), Asp (B3), Arg (B31), Arg (B32) - NH 2 - human insulin, Arg (AO), His (A8), Gly (A21), Asp (B3), Arg (B31), Lys (B32) - NH 2 - human insulin, Arg (AO), His (A8), Gly (A21), Glu (B4), Arg (B31), Arg (B32) - NH 2 - human insulin, Arg (AO), His (A8), Gly (A21), Glu (B4), Arg (B31), Lys (B32) - NH 2 - human insulin, Arg (AO), His (A8), Gly (A21), Glu (BO), Arg (B31), Arg (B32) - NH 2 - human insulin, Arg (AO), His (A8), Gly (A21), GIu (BO ), Arg (B31), Lys (B32) - NH 2 - human insulin, Arg (AO), His (A8), Gly (A21), Arg (B30) - NH 2 - human insulin,
Arg (AO), His (A8), GIy (A21 ), Lys (B30) - NH2 - Humaninsulin, Arg (AO), His (A8), Gly (A21), Lys (B30) - NH 2 - human insulin,
Arg (A-1 ), Arg (AO), His (A8), GIy (A21 ), Arg (B30) - NH2 - Humaninsulin, Arg (A-1), Arg (AO), His (A8), Gly (A21), Arg (B30) - NH 2 - human insulin,
Arg (A-1 ), Arg (AO), His (A8), GIy (A21 ), Lys (B30) - NH2 - Humaninsulin, Arg (A-1), Arg (AO), His (A8), Gly (A21), Lys (B30) - NH 2 - human insulin,
Arg (AO), Arg (A1 ), His (A8), GIy (A21 ), Arg (B30) - NH2 - Humaninsulin, Arg (AO), Arg (A1), His (A8), Gly (A21), Arg (B30) - NH 2 - human insulin,
Arg (AO), Arg (A1 ), His (A8), GIy (A21 ), Lys (B30) - NH2 - Humaninsulin, Arg (AO), Arg (A1), His (A8), Gly (A21), Lys (B30) - NH 2 - human insulin,
His (A8), GIy (A21 ), Arg (B31 ), Arg (B32) - NH2 - Humaninsulin. His (A8), Gly (A21), Arg (B31), Arg (B32) - NH 2 - human insulin.
10. Verfahren gemäß einem oder mehreren der Ansprüche 1 bis 3, wobei das 10. The method according to one or more of claims 1 to 3, wherein the
Insulinderivat ausgewählt wird aus einer Gruppe enthaltend B29-N-myristoyl-des(B30) Humaninsulin, B29-N-palmitoyl-des(B30) Humaninsulin, B29-N-myhstoyl Insulin derivative is selected from a group containing B29-N-myristoyl-des (B30) human insulin, B29-N-palmitoyl-des (B30) human insulin, B29-N-myhstoyl
Humaninsulin, B29-N-palmitoyl Humaninsulin, B28-N-myhstoyl LysB28ProB29 Human insulin, B29-N-palmitoyl human insulin, B28-N-myhstoyl Lys B28 Pro B29
Humaninsulin, B28-N-palmitoyl-LysB28ProB29 Humaninsulin, B30-N-myristoyl-Human insulin, B28-N-palmitoyl-Lys B28 Pro B29 human insulin, B30-N-myristoyl-
ThrB29LysB30 Humaninsulin, B30-N-palmitoyl- ThrB29LysB30 Humaninsulin, B29-N-(N- palmitoyl-Y-glutamyl)-des(B39) Humaninsulin, B29-N-(N-lithocholyl-Y-glutamyl)- des(B30) Humaninsulin, B29-N-(ω-carboxyheptadecanoyl)-des(B30) Humaninsulin and B29-N-(ω-carboxyheptadecanoyl) Humaninsulin. Thr B29 Lys B30 human insulin, B30-N-palmitoyl-Thr B29 Lys B30 human insulin, B29-N- (N-palmitoyl-Y-glutamyl) -des (B39) human insulin, B29-N- (N-lithocholyl-Y-glutamyl ) - of (B30) human insulin, B29-N- (ω-carboxyheptadecanoyl) des (B30) human insulin and B29-N- (ω-carboxyheptadecanoyl) human insulin.
11. Verfahren gemäß einem oder mehreren der Ansprüche 1 bis 10, wobei in der Formulierung ein Konservierungsmittel ausgewählt aus einer Gruppe enthaltend Phenol, m-Cresol, Chlorkresol, Benzylalkohol, Parabene vorhanden ist. 11. The method according to one or more of claims 1 to 10, wherein in the formulation a preservative selected from a group containing phenol, m-cresol, chlorocresol, benzyl alcohol, parabens is present.
12. Verfahren gemäß einem der Ansprüche 1 bis 11 , wobei in der Formulierung ein Isotonisierungsmittel ausgewählt aus einer Gruppe enthaltend Mannitol, Sorbitol,12. The method according to any one of claims 1 to 11, wherein in the formulation an isotonizing agent selected from a group containing mannitol, sorbitol,
Lactose, Dextrose, Trehalose, Natriumchlorid, Glycerol vorhanden ist. Lactose, dextrose, trehalose, sodium chloride, glycerol is present.
13. Verfahren gemäß einem der Ansprüche 1 bis 12, wobei das Insulin, das 13. The method according to any one of claims 1 to 12, wherein the insulin, the
Insulinanalogon und/oder das Insulinderivat in einer Konzentration von 240 - 3000 nmol/ml vorliegt Insulin analog and / or the insulin derivative in a concentration of 240-3000 nmol / ml is present
14. Verfahren gemäß einem oder mehreren der Ansprüche 1 bis 13, wobei in der Formulierung 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. The method according to one or more of claims 1 to 13, wherein in the formulation additionally a glucagon-Like Peptide-1 (GLP1) or an analog or derivative thereof, or exendin-3 or -4 or an analog or derivative thereof is.
15. Verfahren gemäß Anspruch 14, bei dem ein Analogon von Exendin-4 ausgewählt wird aus einer Gruppe enthaltend 15. The method of claim 14, wherein an analog of exendin-4 is selected from a group comprising
H-desPro36-Exendin-4-Lys6-NH2, H-desPro 36 -Exendin-4-Lys 6 -NH 2 ,
H-des(Pro36 37)-Exendin-4-l_ys4-NH2 und H des (Pro 36 37 ) -Exendin-4-l_ys 4 -NH 2 and
H-des(Pro36 37)-Exendin-4-Lys5-NH2, H des (Pro 36 37 ) -Exendin-4-Lys 5 -NH 2 ,
oder ein pharmakologisch tolerierbares Salz davon. or a pharmacologically tolerable salt thereof.
16. Verfahren gemäß Anspruch 14, bei dem ein Analogon von Exendin-4 ausgewählt wird aus einer Gruppe enthaltend The method of claim 14, wherein an analog of exendin-4 is selected from a group comprising
desPro36 [Asp28] Exend in-4 (1 -39), desPro 36 [Asp 28 ] Exend in-4 (1 -39),
desPro36 [IsoAsp28] Exend in-4 (1 -39), desPro36 [Met(O)14, Asp28] Exend in-4 (1 -39), desPro 36 [IsoAsp 28 ] Exend in-4 (1 -39), DESpro 36 [Met (O) 14, Asp 28] exend in-4 (1 -39)
desPro36 [Met(O)14, IsoAsp28] Exend in-4 (1 -39), desPro 36 [Met (O) 14 , IsoAsp 28 ] Exend in-4 (1 -39),
desPro36 [Trp(O2)25, Asp28] Exend in-2 (1 -39), desPro 36 [Trp (O 2 ) 25 , Asp 28 ] Exend in-2 (1 -39),
desPro36 [Trp(O2)25, IsoAsp28] Exend in-2 (1 -39), desPro 36 [Trp (O 2 ) 25 , IsoAsp 28 ] Exend in-2 (1 -39),
desPro36 [Met(O)14Trp(O2)25, Asp28] Exend in-4 (1 -39) und desPro 36 [Met (O) 14 Trp (O 2 ) 25 , Asp 28 ] Exend in-4 (1 -39) and
desPro36 [Met(O)14Trp(O2)25, IsoAsp28] Exend in-4 (1 -39), desPro 36 [Met (O) 14 Trp (O 2 ) 25 , IsoAsp 28 ] Exend in-4 (1 -39),
oder ein pharmakologisch tolerierbares Salz davon. or a pharmacologically tolerable salt thereof.
17. Verfahren gemäß Anspruch 16, bei denen an die C-Termini der Analoga von Exendin-4 das Peptid -LyS6-NH2 angefügt ist. 17. The method according to claim 16, wherein to the C-termini of the analogs of exendin-4 the peptide -LyS 6 -NH 2 is added.
18. 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-(LyS)6- des Pro36 [Asp28]Exendin-4(1 -39)-Lys6-NH2 H- (LyS) 6 - of Pro 36 [Asp 28 ] Exendin-4 (1 -39) -Lys 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] Exend in-4(1 -39) -NH2, H-Asn- (Glu) 5 of the Pro 36 , Pro 37 , Pro 38 [Asp 28 ] Exend in-4 (1 -39) -NH 2 ,
des Pro36, Pro37, Pro38 [Asp28]Exendin-4(1 -39)-(Lys)6-NH2, Pro 36 , Pro 37 , Pro 38 [Asp 28 ] Exendin-4 (1 -39) - (Lys) 6 -NH 2 ,
H-(LyS)6- des Pro36, Pro37, Pro38 [Asp28]Exendin-4(1 -39)-(Lys)6-NH2, H- (LyS) 6 - of Pro 36 , Pro 37 , Pro 38 [Asp 28 ] Exendin-4 (1 -39) - (Lys) 6 -NH 2 ,
H-Asn-(Glu)5- des Pro36, Pro37, Pro38 [Asp28] Exend in-4(1 -39)-(Lys)6-NH2, H-Asn- (Glu) 5 - of the Pro 36 , Pro 37 , Pro 38 [Asp 28 ] Exend in-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] Exend in-4(1 -39) -NH2, H- the Asp 28 Pro 36, Pro 37, Pro 38 [Trp (O2) 25] exend in-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-(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, 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, Asp28]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 , 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-(Glu)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 , 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- (LyS) 6 - of Pro 36 , Pro 37 , Pro 38 [Met (O) 14 , Asp 28 ] Exendin-4 (1 -39) -Lys 6 -NH 2 ,
H-ASn-(GIu)5 des Pro36, Pro37, Pro38 [Met(O)14, Asp28] Exendin-4(1 -39)-(Lys)6-NH2, H-(LyS)6- des Pro36 [Met(O)14, Trp(O2)25, Asp28]Exendin-4(1 -39)-Lys6-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 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, of Asp 28 Pro 36, Pro 37, Pro 38 [Met (O) 14, Trp (O2) 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 - 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- des Pro36' Pro37, Pro38 [Met(O)14, Trp(O2)25, Asp28] Exend in-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 ] Exend in-4 (1 -39) - (Lys) 6 - NH 2 .
H-Asn-(Glu)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.
19. Verfahren nach Anspruch 14, wobei in der Formulierung zusätzlich Arg34, Lys26 (Nε(γ-glutamyl(Nα-hexadecanoyl))) GLP-1 (7-37) [liraglutide] oder ein pharmakologisch tolerierbares Salz davon enthalten ist. The method of claim 14, wherein the formulation additionally contains Arg 34 , Lys 26 (N ε (γ-glutamyl (N α -hexadecanoyl)), GLP-1 (7-37) [liraglutide] or a pharmacologically tolerable salt thereof is.
20. Verfahren nach einem oder mehreren der Ansprüche 1 bis 19, wobei in der Formulierung noch ein Zinksalz enthalten ist. 20. The method according to one or more of claims 1 to 19, wherein in the formulation nor a zinc salt is included.
21. Verwendung eines Verfahrens nach einem oder mehreren der Ansprüche 1 bis 20 bei der großtechnischen Herstellung eines Insulins, Insulinanalogs oder 21. Use of a method according to one or more of claims 1 to 20 in the large-scale production of an insulin, insulin analog or
Insulinderivats. Insulin derivative.
22. Zweiteiliger Satz von Behältern, in denen einer der Behälter ein Insulin, ein 22. Two-part set of containers in which one of the containers contains an insulin
Insulinanalogon oder ein Insulinderivat als Feststoff und der andere Behälter ein Lösemittelgemisch eines bestimmten pH-Werts mit der Endkonzentration der Exzipienten einer gewünschten Formulierung eines Insulins, eines Insulinanalogons oder eines Insulinderivats enthält; zur hitze- und schüttelstabilen Aufbewahrung des Insulins, des Insulinanalogons oder des Insulinderivats für das spätere Herstellen einer gebrauchsfertigen Formulierung durch Lösen des Feststoffs in dem Lösemittelgemisch gemäß einem oder mehreren der Ansprüche 1 bis 20. Insulin analogue or an insulin derivative as solid and the other container containing a solvent mixture of a particular pH with the final concentration of the excipients of a desired formulation of an insulin, an insulin analog or an insulin derivative; for heat- and shake-stable storage of insulin, the insulin analog or the insulin derivative for the subsequent production of a ready-to-use formulation by dissolving the solid in the solvent mixture according to one or more of claims 1 to 20.
23. Zweikammerinjektionssystem bei dem eine Kammer ein Insulin, ein 23. Two-chamber injection system in which a chamber insulin, a
Insulinanalogon oder ein Insulinderivat als Feststoff und die andere Kammer ein Lösemittelgemisch eines bestimmten pH-Werts mit der Endkonzentration der Exzipienten einer gewünschten Formulierung eines Insulins, eines Insulinanalogons oder eines Insulinderivats enthält; zur hitze- und schüttelstabilen Aufbewahrung des Insulins, des Insulinanalogons oder des Insulinderivats für das spätere Herstellen einer gebrauchsfertigen Formulierung durch Lösen des Feststoffs in dem Lösemittelgemisch gemäß einem oder mehreren der Ansprüche 1 bis 20. Insulin analogue or an insulin derivative as a solid and the other chamber containing a solvent mixture of a particular pH with the final concentration of the excipients of a desired formulation of an insulin, an insulin analog or an insulin derivative; for the heat- and shake-stable storage of the insulin, the insulin analog or the insulin derivative for the subsequent production of a ready-to-use formulation by dissolving the solid in the solvent mixture according to one or more of claims 1 to 20.
EP10730441A 2009-07-06 2010-07-02 Heat- and vibration-stable insulin preparations Withdrawn EP2451472A1 (en)

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