Rapid-acting insulin compositions

Abstract

To provide a composition of human insulin or insulin analog that has faster pharmacokinetic and / or pharmacodynamic action than commercial formulations of existing insulin analog products.SOLUTION: A pharmaceutical composition comprises (a) insulin, (b) citrate in a concentration of about 10 to about 30 mM, (c) zinc in a concentration of about 0.2 to about 2 mM, (d) magnesium in a concentration of about 1 to about 15 mM, (e) total chloride in a concentration of about 10 to about 60 mM, (f) a surfactant in a concentration of about 0.001 to about 0.2% w / v, and (g) a preservative.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a method for treating diabetes mellitus by combating postprandial glycemic excursions and treating the acute phase of hyperglycemia. A pharmaceutical composition for the treatment of insulin. The composition optionally contains sodium chloride. , zinc and optionally magnesium chloride and / or a surfactant. The composition contains a phosphorus or insulin analogue, and specific concentrations of citrate and chloride. The insulin composition has a faster uptake of insulin from the injection site than existing commercially available insulin compositions. The composition is used to reduce insulin activity during meals or to reduce hyperglycemia when insulin is needed. It is useful in providing acute care. [Background technology]

[0002] The time / action profile of insulin is important in controlling postprandial blood glucose levels. In healthy people, the pancreas responds to absorbed food by secreting spikes of insulin, which This causes a rise in blood insulin levels within minutes. In some people, insulin must be administered. Phosphorus enters the blood slowly. Inadequate release of insulin and too late at the start of a meal Onset of effect is hyperglycemia during or immediately after meals. Excessive insulin during meals can cause postprandial hypoglycemia.

[0003] There have been previous efforts to reduce the time / action of insulin products. Early efforts to address this issue include insulin lispro (HUMALOG®), insulin Aspart (NOVOLOG®) and insulin glulisine (APID These included the development of novel "fast-acting" insulin analogs such as insulin RA. Insulin lyspro and insulin aspart are similar to the amino acid chain arrangement in human insulin. It achieves its immediate effect through a change in the amino acid sequence from the insulin glulisine. It contains a change in the amino acid sequence of human insulin, but also lacks zinc and is stable. Rapid-acting insulin analogs were developed in the 1990s and early 2000s. However, even the so-called rapid-acting insulin analogues have Maximum insulin levels are not reached until 50-90 minutes after the pancreas is functioning normally. slower than insulin released by the liver, matching the carbohydrate absorption profile. is not limited to this.

[0004] Another means of achieving rapid action that is being explored is to provide it in combination with insulin. One such approach is the use of ingredients or excipients that improve the time-action profile of insulin.

[0005] For example, US2013 / 0231281 describes The present invention discloses an aqueous composition containing insulin and specific oligosaccharides in combination. The active compounds are dextran methyl carboxylic acid, polyglutamic acid, polyaspartic acid, Based on PAA (polyacrylic acid), alginate, hyaluronic acid, glucuronic acid or An anionic polymer selected from the group consisting of polymers based on galacturonic acid and its salts. -, or citric acid, aspartic acid, glutamic acid, malic acid, tartaric acid, succinic acid, From the group consisting of dipic acid, oxalic acid, triphosphonates, polyphosphonates and their salts In one embodiment, the polyanionic compound is selected from the group consisting of citric acid, glycerol, tert-butyl ether ... The polyanionic compound is a single oligosaccharide or a polyanionic compound. Improves the performance of oligosaccharide-containing compositions even when the compound alone has no effect on time-dependent effects. It is said that the insulin concentration in the composition containing citrate but lacking oligosaccharides is No time-effect results for phosphorus were disclosed.

[0006] In addition, US2012 / 0178675 and US2014 / 0113856, even if For example, zinc chelators such as ethylenediaminetetraacetic acid (EDTA) and citric acid In combination with what are described as "solubilizing / stabilizing" agents such as acid or sodium citrate In addition, US2014 / 0113856 discloses a composition containing insulin. "Early clinical trials of such compositions have shown discomfort at the injection site," Magne said. It is claimed that the addition of a calcium-containing compound increases tolerance at the injection site. Magnesium compounds include inorganic salts such as magnesium hydroxide, magnesium sulfate, Magnesium chloride, magnesium bromide, magnesium iodide, magnesium pyrophosphate, Magnesium sulfate heptahydrate and magnesium oxide; and organic salts such as magnesium EDTA, magnesium lactate, magnesium aspartate, magnesium acetate, magnesium carbonate US2 includes magnesium, magnesium citrate, and magnesium gluconate. US2012 / 0178675 and US2014 / 0113856 are directed to citric acid or The insulin time action performance for the salt-containing but EDTA-free composition was However, a 2012 poster by the designated attorneys for those applications Time action of insulin lispro compositions containing citrate but not EDTA The profile was compared to compositions containing various concentrations of EDTA and citrate. Starr said, "The citrate salt alone in the formulation does not alter the absorption profile of [insulin lispro]. "The chelation of zinc with EDTA (which is not very stable) is insufficient to improve (accelerates the disassembly of insulin analog hexamers) and citrate (masks the surface charge and prevents reaggregation) "Both the stimulant and the inhibitory effect (which inhibits absorption) are required above a threshold concentration that enhances the subcutaneous rate of absorption," he said. http: / / files.shareholder.com / download s / BIOD / 3315705516x0x602912 / 3C955886-6AA4 -4D66-BD33-3FFB4C906B25 / EASD_Poster_Sept Roderike Poh, available at ember_2012_FINAL.pdf l et al., “Development of Ultra-Rapid-Acting P randial Insulin Analogs Requires Chelati on of Zinc Ions and Charge Masking to In crease the Rate of Subcutaneous Absorpti on”.

[0007] US2015 / 0065423 discloses a method for treating a patient suffering from a vascular endothelial disorder comprising administering to said patient a peptide and a vasodilator (e.g., nitroglycerin). and optionally what is described as a "charge masking agent," e.g., citrate, diketone, Piperazine, diketopiperazine derivatives, EDTA, diarginine piperazine, diarginine Diarginine piperazine salts, diarginine piperazine isomers, diarginine piperazine esters and The present invention describes compositions containing any combination thereof. Compositions lacking an extender are not disclosed.

[0008] The prior art collectively describes ultrafast acting insulin preparations and their use with excipients included in the present invention. does not teach a specific concentration of

[0009] Despite these efforts, there is still a high risk of infection from injection sites due to the fact that these drugs are intended for use with meals. More rapid uptake of insulin; faster onset of action than existing insulin products; and There remains a need for insulin compositions that have chemical and physical stability during storage. The present invention seeks to provide compositions that meet these needs. Summary of the Invention

[0010] The inventors have surprisingly found that compositions containing specific concentrations of citrate are more effective than existing commercially available and more rapid uptake and / or onset of action of insulin in the blood than the insulin composition of Without excluding the improvement of time action, for example, zinc, Certain concentrations of sodium chloride, and optionally magnesium chloride and / or surfactants By incorporating a stabilizer into the composition, the chemical and physical properties of the composition can be improved under certain conditions. Stability is maintained.

[0011] Thus, the present invention relates to a method for treating a patient having an insulin deficiency, comprising administering to said patient an insulin deficiency at a concentration of about 100 to about 200 IU / mL and administering to said patient an insulin deficiency at a concentration of about 15 to about 200 IU / mL. citrate at a concentration of about 35 mM, zinc at a concentration of about 0.2 to about 0.8 mM, and a preservative In certain embodiments, the pharmaceutical composition further comprises up to about 5 mM The composition further comprises magnesium chloride at a concentration of

[0012] In another embodiment, the present invention generally relates to a pharmaceutical composition comprising insulin, a particular concentration of citrate, and a stabilizer. A composition comprising:

[0013] In a particular embodiment, the present invention provides a method for the preparation of a medicament for the treatment of a disease comprising administering to said patient insulin and citric acid at a concentration of about 10 to about 30 mM. a salt of zinc at a concentration of about 0.2 to about 2 mM and a salt of magnesium at a concentration of about 1 to about 15 mM. and total chloride at a concentration of about 10 to about 60 mM and about 0.001 to about 0.2% w / v. and a preservative.

[0014] In a particular embodiment, the present invention provides a method for the preparation of a medicament for the preparation of a pharmaceutical composition comprising insulin and citric acid at a concentration of about 10 to about 30 mM. a salt of zinc at a concentration of about 0.2 to about 2 mM and a salt of magnesium at a concentration of about 1 to about 15 mM. and sodium chloride at a concentration of about 1 to about 50 mM and about 0.001 to about 0.2% w / v and a preservative.

[0015] In certain embodiments, the pharmaceutical compositions of the present invention are provided at a concentration of about 100 to about 500 IU / mL. of insulin lispro, citrate at a concentration of about 10 to about 25 mM, and about 2 to about 9 mM magnesium at a concentration of about 0.3 to about 1.3 mM, zinc at a concentration of about 15 to about 35 mM, Total chloride at a concentration of 1 mM and poloxamer 1 at a concentration of about 0.03 to about 0.12% w / v. 88 and meta-cresol at a concentration of about 2.8 to about 3.5 mg / mL, The pH of the product is approximately 7.0 to 7.8.

[0016] In certain embodiments, the pharmaceutical compositions of the present invention are provided at a concentration of about 100 to about 500 IU / mL. of insulin lispro, citrate at a concentration of about 10 to about 25 mM, and about 2 to about 9 mM magnesium at a concentration of about 0.3 to about 1.3 mM, zinc at a concentration of about 1 to about 25 mM Sodium chloride at a concentration of about 0.03 to about 0.12% w / v and poloxamer at a concentration of about 0.03 to about 0.12% w / v. and meta-cresol at a concentration of about 2.8 to about 3.5 mg / mL, The pH of the composition is about 7.0 to 7.8.

[0017] In certain embodiments, the pharmaceutical compositions of the present invention are provided at a concentration of about 100 to about 200 IU / mL. of insulin lispro, citrate at a concentration of about 15 to about 25 mM, and about 3 to about 8 mM magnesium chloride at a concentration of about 0.3 to about 1.1 mM, zinc at a concentration of about 20 to about Total chloride at a concentration of 25 mM and poloxamer at a concentration of about 0.06 to about 0.09% w / v. and meta-cresol at a concentration of about 2.8 to about 3.5 mg / mL, The pH of the composition is about 7.0 to 7.8.

[0018] In certain embodiments, the pharmaceutical compositions of the present invention are provided at a concentration of about 100 to about 200 IU / mL. of insulin lispro, citrate at a concentration of about 15 to about 25 mM, and about 3 to about 8 mM magnesium chloride at a concentration of about 0.3 to about 1.1 mM, zinc at a concentration of about 1 to about 2 Sodium chloride at a concentration of about 0.001 mM and poloxamer at a concentration of about 0.06 to about 0.09% w / v. and meta-cresol at a concentration of about 2.8 to about 3.5 mg / mL. At this time, the pH of the composition is about 7.0 to 7.8.

[0019] In certain embodiments, the pharmaceutical compositions of the present invention are provided at a concentration of about 100 to about 200 IU / mL. of insulin lispro, citrate at a concentration of about 15 to about 25 mM, and about 3 to about 8 mM magnesium chloride at a concentration of about 0.6 to about 1.1 mM, zinc at a concentration of about 20 to about Total chloride at a concentration of 25 mM and poloxamer at a concentration of about 0.06 to about 0.09% w / v. and meta-cresol at a concentration of about 2.8 to about 3.5 mg / mL, The pH of the composition is about 7.0 to 7.8.

[0020] In certain embodiments, the pharmaceutical compositions of the present invention are provided at a concentration of about 100 to about 200 IU / mL. of insulin lispro, citrate at a concentration of about 15 to about 25 mM, and about 3 to about 8 mM magnesium chloride at a concentration of about 0.6 to about 1.1 mM, zinc at a concentration of about 1 to about 2 Sodium chloride at a concentration of about 0.001 mM and poloxamer at a concentration of about 0.06 to about 0.09% w / v. and meta-cresol at a concentration of about 2.8 to about 3.5 mg / mL. At this time, the pH of the composition is about 7.0 to 7.8.

[0021] In a particular embodiment, the pharmaceutical composition of the present invention contains insulin at a concentration of 100 IU / mL. Lispro, citrate at a concentration of about 25 mM, and subtilisin at a concentration of about 0.6 to about 0.9 mM. Lead, magnesium chloride at a concentration of about 5 mM, and total chloride at a concentration of about 20 to about 25 mM and poloxamer 188 at a concentration of about 0.09% w / v and poloxamer 188 at a concentration of about 3.15 mg / mL. and meta-cresol at 100° C., wherein the pH of the composition is about 7.4.

[0022] In a particular embodiment, the pharmaceutical composition of the present invention contains insulin at a concentration of 100 IU / mL. Lispro, citrate at a concentration of about 25 mM, and subtilisin at a concentration of about 0.6 to about 0.9 mM. Lead, magnesium chloride at a concentration of about 5 mM, and sodium chloride at a concentration of about 1 to about 20 mM. poloxamer 188 at a concentration of about 0.09% w / v and poloxamer 188 at a concentration of about 3.15 mg / mL. and meta-cresol in a concentration, wherein the pH of the composition is about 7.4.

[0023] In a particular embodiment, the pharmaceutical composition of the present invention contains insulin at a concentration of 100 IU / mL. Lispro, citrate at a concentration of about 25 mM, zinc at a concentration of about 0.9 mM, and Magnesium chloride at a concentration of about 100 mM, total chloride at a concentration of about 20 to about 25 mM, and about 0. Poloxamer 188 at a concentration of 0.09% w / v and methacrylamide at a concentration of approximately 3.15 mg / mL. and a resole, wherein the pH of the composition is about 7.4.

[0024] In a particular embodiment, the pharmaceutical composition of the present invention contains insulin at a concentration of 100 IU / mL. Lispro, citrate at a concentration of about 25 mM, zinc at a concentration of about 0.9 mM, and magnesium chloride at a concentration of about 1 to about 20 mM, sodium chloride at a concentration of about 1 to about 20 mM, and Poloxamer 188 at a concentration of 0.09% w / v and methymol at a concentration of approximately 3.15 mg / mL. and tacresol, wherein the pH of the composition is about 7.4.

[0025] In a particular embodiment, the pharmaceutical composition of the present invention contains insulin at a concentration of about 200 IU / mL. and citrate at a concentration of about 15 to about 25 mM and citrate at a concentration of about 3 to about 8 mM. Magnesium chloride, zinc at a concentration of about 0.8 to about 1.1 mM, and about 20 to about 25 mM and poloxamer 188 at a concentration of about 0.06 to about 0.09% w / v. and meta-cresol in a concentration of about 2.8 to about 3.5 mg / mL, wherein the composition The pH is approximately 7.0 to 7.8.

[0026] In a particular embodiment, the pharmaceutical composition of the present invention contains insulin at a concentration of about 200 IU / mL. and citrate at a concentration of about 15 to about 25 mM and citrate at a concentration of about 3 to about 8 mM. Magnesium chloride, zinc at a concentration of about 0.8 to about 1.1 mM, and zinc at a concentration of about 1 to about 20 mM. and poloxamer 18 at a concentration of about 0.06 to about 0.09% w / v. 8 and meta-cresol at a concentration of about 2.8 to about 3.5 mg / mL, The pH of the substance is approximately 7.0 to 7.8.

[0027] In addition, the present invention also provides a method for administering an effective dose of the pharmaceutical composition of the present invention to a human in need thereof. Also provided is a method for treating diabetes comprising:

[0028] In addition, the present invention provides pharmaceutical compositions of the present invention for use in therapy. In particular, the present invention provides a pharmaceutical composition of the present invention for use in the treatment of diabetes. Ming also discloses the use of the pharmaceutical composition of the present invention in producing a drug for the treatment of diabetes. Also provided.

[0029] Additionally, the present invention provides an article of manufacture comprising the pharmaceutical composition of the present invention. An article of manufacture is provided that includes the pharmaceutical composition of the present invention, wherein the article of manufacture is a multi-use vial. In particular, the present invention relates to a pharmaceutical composition comprising the pharmaceutical composition of the present invention, wherein the article of manufacture is a reusable pen-type injector. In particular, the present invention provides an article of manufacture comprising a poultice for continuous subcutaneous insulin infusion therapy. An article of manufacture comprising the pharmaceutical composition of the invention is provided which is a pump device.

[0030] In embodiments, the insulin is human insulin or, for example, insulin lysprine. b) Rapid-acting human insulin, such as insulin aspart or insulin glulisine A variant, mutant or analogue of the above structure is selected from the group consisting of: In some embodiments, the insulin is insulin lispro.

[0031] In an embodiment, the preservative is selected from the group consisting of phenol and meta-cresol. Preferably, the preservative is meta-cresol. In an embodiment, the concentration of meta-cresol is about 2 Preferably, the concentration of meta-cresol is 3.5 mg / mL to about 3.8 mg / mL. It is 15mg / mL.

[0032] In an embodiment, the composition further comprises a tonicity agent. In an embodiment, the tonicity agent is glycerol. In certain embodiments, the concentration of glycerol is about 1 to about 16 mg / mL.

[0033] In embodiments, the composition further comprises a buffer. In certain embodiments, the buffer is sodium phosphate. It is thorium.

[0034] In certain embodiments, the composition further comprises an additional stabilizer. The article further comprises a surfactant.

[0035] In certain embodiments, the composition may further comprise an additional chelating agent, such as, for example, EDTA, nitrile, It does not contain additional vasodilators such as roglycerin and / or oligosaccharides.

[0036] In certain preferred embodiments, the pH of the composition is about 7.0 to about 7.8. In a particularly preferred embodiment, the pH of the composition is about 7.3 to about 7.5. The pH is approximately 7.4.

[0037] In certain embodiments, one or more pharmacokinetic or pharmacodynamic parameters related to time action are For example, time to maximum insulin concentration (Tmax); Half-reach time (initial 1 / 2Tmax); the time to half-reach of insulin during the downward phase of the concentration versus time curve. Time to reach half of maximum concentration (late 1 / 2Tmax); between early and late 1 / 2Tmax time (Tmax spread); different time points based on the minute area under the insulin concentration curve The proportion of the total insulin dose absorbed over time (e.g., AUC 0-30分 , AUC 0- 60分 , AUC 0-120分 , AUC 0-180分 ); half of the total insulin concentration Time to reach maximum glucose infusion rate (GIRmax); time to reach maximum glucose infusion rate (T50); time to reach half of the infusion rate (initial 1 / 2 GIRmax); time to reach the peak Time to reach half of the maximum glucose infusion rate (late 1 / 2GIRmax); GIR curve The proportion of total glucose infused at different times based on the minute area under the line (e.g. , G.I.R. 0-30分 , G.I.R. 0-60分 , G.I.R. 0-120分 , G.I.R. 0-180分 ) When measured in this way, the pharmaceutical composition is a citrate-free, fast-acting insulin analog. At least 10%, at least 20%, at least 30%, at least 40% or at least 50% more rapid uptake of insulin into the blood and / or Provides onset of action.

[0038] In certain embodiments, the pharmaceutical composition comprises a compound having a time-action profile, such as those described above. citrate as measured by one or more associated pharmacokinetic or pharmacodynamic parameters Between 10% and 50% faster than salt-free rapid-acting insulin analogue compositions Provides insulin uptake into the blood and / or onset of action.

[0039] In certain embodiments, the pharmaceutical composition is capable of being stored at 2-8° C. for at least 24 months. In certain embodiments, the pharmaceutical composition is in a reusable pen injection device. For vials or cartridges up to 28 days of use at temperatures up to 30°C In certain embodiments, the pharmaceutical composition is stable for at least 36 months at 2-8°C. In certain embodiments, the pharmaceutical composition is reusable. For vials or cartridges in pen injectors, the product can be stored for up to 32 days at temperatures up to 30°C. In certain embodiments, the composition is stable enough to allow for use at room temperature for up to 7 days. It is stable enough to allow its use in a pump device for continuous subcutaneous insulin infusion therapy.

[0040] In certain embodiments, the pharmaceutical composition comprises an insulin inhibitor at a concentration of about 100 IU / mL. citrate at a concentration of about 15 mM; zinc at a concentration of about 0.3 mM; sodium chloride at a concentration of about 5 mM, magnesium chloride at a concentration of about 3.15 mg glycerol at a concentration of about 7.6 mg / mL. .

[0041] In another embodiment, the pharmaceutical composition comprises insulin lisprosulfone at a concentration of about 100 IU / mL. citrate at a concentration of about 25 mM; zinc at a concentration of about 0.3 mM; sodium chloride at a concentration of about 5 mM, magnesium chloride at a concentration of about 3.15 mg / The mixture contains metacresol at a concentration of about 1000 mg / mL and glycerol at a concentration of about 4.5 mg / mL.

[0042] In another embodiment, the pharmaceutical composition comprises insulin lisprosulfone at a concentration of about 100 IU / mL. citrate at a concentration of about 35 mM; zinc at a concentration of about 0.3 mM; sodium chloride at a concentration of about 5 mM, magnesium chloride at a concentration of about 3.15 mg / and meta-cresol at a concentration of 1.0 mL.

[0043] In another embodiment, the pharmaceutical composition comprises insulin lisprosulfone at a concentration of about 100 IU / mL. citrate at a concentration of about 25 mM; zinc at a concentration of about 0.3 mM; and meta-cresol at a concentration of about 3.15 mg / mL.

[0044] In another embodiment, the pharmaceutical composition comprises insulin lisprosulfone at a concentration of about 100 IU / mL. citrate at a concentration of about 15 mM; zinc at a concentration of about 0.3 mM; sodium chloride at a concentration of about 5 mM, magnesium chloride at a concentration of about 3.15 mg / 6.6 mg / mL of metacresol and glycerol at a concentration of about 7.6 mg / mL, In this regard, the pharmaceutical composition may comprise one or more time-related agents, such as those described above. Citrate-free as measured by pharmacokinetic or pharmacodynamic parameters Between 10% and 20%, between 20% and 30%, and between 3% and 4% of the composition of fast-acting insulin analogues. Insulin uptake into the blood is between 0% and 40% or between 40% and 50% more rapid and / or provide an onset of action.

[0045] In another embodiment, the pharmaceutical composition comprises insulin lisprosulfone at a concentration of about 100 IU / mL. citrate at a concentration of about 25 mM; zinc at a concentration of about 0.3 mM; sodium chloride at a concentration of about 5 mM, magnesium chloride at a concentration of about 3.15 mg / The mixture contained metacresol at a concentration of about 4.5 mg / mL and glycerol at a concentration of about 4.5 mg / mL. wherein the pharmaceutical composition comprises one or more of the above-described time-related actions. citrate-free as measured by pharmacokinetic or pharmacodynamic parameters The insulin analogues are absorbed into the bloodstream 10% to 50% more quickly than the fast-acting insulin analogues. The present invention provides for the uptake and / or onset of action of thorin.

[0046] In another embodiment, the pharmaceutical composition comprises insulin lisprosulfone at a concentration of about 100 IU / mL. citrate at a concentration of about 35 mM; zinc at a concentration of about 0.3 mM; sodium chloride at a concentration of about 5 mM, magnesium chloride at a concentration of about 3.15 mg / and meta-cresol in a concentration of 100 mL / ml, wherein the pharmaceutical composition is as described above. by one or more pharmacokinetic or pharmacodynamic parameters related to time action, such as When measured, it was 10 times more effective than a non-citrate-containing rapid-acting insulin analogue composition. 100%-50% faster insulin uptake into the bloodstream and / or onset of action do.

[0047] In another embodiment, the pharmaceutical composition comprises insulin lisprosulfone at a concentration of about 100 IU / mL. citrate at a concentration of about 25 mM; zinc at a concentration of about 0.3 mM; and meta-cresol at a concentration of about 3.15 mg / mL, In this regard, the pharmaceutical composition may comprise one or more time-related agents, such as those described above. Citrate-free as measured by pharmacokinetic or pharmacodynamic parameters The insulin is released into the bloodstream 10% to 50% more quickly than fast-acting insulin analogues. Provides phosphorus uptake and / or onset of action. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0048] As used herein, the term "composition" refers to a mixture of insulin and other ingredients or excipients. Combinations of insulin and other ingredients or excipients in a single combined preparation It refers to the combination.

[0049] As used herein, "insulin" refers to human insulin or human insulin A rapid-acting form of human insulin that has the functional activity of but a faster onset of action. A structural variant, mutant or analogue of human insulin. The bodies are insulin lispro, insulin aspart, and insulin glulisine. Insulin for commercial products is produced using recombinant DNA technology or by chemical synthesis. Recombinant methods are well known and are highly preferred. The molecule of human insulin (CAS no. 11061-68-0) consists of two amino acid chains A and B, the sequence of which is known. .

[0050] The human insulin A chain has the following sequence of amino acids: GlyIleValGluGlnCysCysThrSerIleCysSerLeuT yrGlnLeuGluAsnTyrCysAsn (SEQ ID NO: 1)

[0051] The human insulin B chain has the following sequence of amino acids: PheValAsnGlnHisLeuCysGlySerHisLeuValGluA laLeuTyrLeuValCysGlyGluArgGlyPhePheTyrTh rProLysThr (SEQ ID NO: 2)

[0052] The chain is bounded by two disulfide bonds: CysA7-CysB7 and CysA20-CysB1. 9. The A chain is connected by an intrachain disulfide bond at CysA6-CysA11. Human insulin has the empirical formula C 257 H 383 N 65 O 77 S 6 and 5808 minutes It has a molecular weight.

[0053] Insulin lispro, the drug substance in HUMALOG®, is ranked 28th and and the natural proline-lysine inversion on the B chain at position 29 (28 B -L-lysine-29 B -L -proline Human insulin) in terms of its primary amino acid sequence Insulin lispro (CAS number 133107-64-9) is the same as It has been shown to be equipotent to human insulin at baseline, but its effectiveness after subcutaneous injection is It is more rapid and shorter-lasting than injected soluble human insulin. The registered trademark contains m-cresol as a preservative, stabilizer, and tonicity adjuster (glycerol). ), buffering agent (dibasic sodium phosphate), stabilizer (zinc oxide) and pH adjustment for the vehicle Contains conditioners.

[0054] The insulin lispro molecule consists of human insulin A chains crosslinked to insulin lispro B chains. (SEQ ID NO: 1), and the amino acid sequence of the insulin lispro B chain is set forth in SEQ ID NO: 3 below. is given by: PheValAsnGlnHisLeuCysGlySerHisLeuValGluA laLeuTyrLeuValCysGlyGluArgGlyPhePheTyrTh rLysProThr (SEQ ID NO: 3)

[0055] The chemical formula for insulin lispro is C 257 H 383 N 65 O 77 S 6 and its molecular weight is approximately 5808. One unit of insulin lispro contains 0.0347 mg of insulin. Equivalent to Lispro.

[0056] The drug substance in NOVOLOG®, insulin aspart (CAS no. No. 116094-23-6) is another rapid-acting insulin analogue. Its structure is The A chain of insulin (SEQ ID NO:1) and B28 as reflected in the amino acid sequence Pro is replaced by Asp (Pro-B28-Asp-human insulin) The B chain consists of: PheValAsnGlnHisLeuCysGlySerHisLeuValGluA laLeuTyrLeuValCysGlyGluArgGlyPhePheTyrTh rAspLysThr (SEQ ID NO: 4)

[0057] Insulin Aspart (28 B Aspartic acid-human insulin) has the empirical formula C 256 H 381 N 65 O79 S 6 and has a molecular weight of about 5826. The concentration is 6 nmoles, which corresponds to 0.035 mg of salt-free anhydrous insulin aspart. Equivalent.

[0058] Insulin glulisine (CAS no. 207748-29-6) is yet another rapid-acting insulin analogue. The lysine molecule is reflected in the human insulin A chain (SEQ ID NO:1) and the following amino acid sequence: The B chain is modified (Asn-B3-Lys, Lys-B2 9-Glu) consisting of: PheValLysGlnHisLeuCysGlySerHisLeuValGluA laLeuTyrLeuValCysGlyGluArgGlyPhePheTyrTh rProGluThr (SEQ ID NO:5)

[0059] Insulin glulisine (3 B -Lysine-29 B -glutamic acid-human insulin) Experiment C 258 H 384 N 64 O 78 S 6 and has a molecular weight of 5823. One unit of lysine approximately corresponds to 0.0349 mg of insulin glulisine.

[0060] The following schemes are currently approved for use in treating prandial variations in blood glucose. The amino acid sequences and disulfide bonds of human insulin and rapid-acting insulin analogs Indicates the combination. Human insulin A chain [ka]

[0061] In a particular embodiment, the compositions of the invention contain about 100 to about 500 IU / mL of insulin. In certain embodiments, the compositions of the present invention have a concentration of about 100 to about 300 IU / mL. In a specific embodiment, the composition of the present invention has a concentration of insulin of about 100 to about 200 IU / mL of insulin. A particular composition contains about 100 IU / mL. A particular composition contains about 200 IU / mL.

[0062] The time-action profiles of the above-referenced insulin analogs demonstrated in this invention The improvement is achieved through the use of certain specific concentrations of citrate, US2012 / 017867 5 and those described in US 2014 / 0113856 (e.g., EDTA). Additional suitable chelating agents include those described in US 2015 / 0065423 (e.g., Additional vasodilators such as glycerin, or This is achieved without the need for other oligosaccharides such as those described.

[0063] Citrate ion has the chemical name 2-hydroxypropane-1,2,3-tricarboxylate , molecular formula C 6 H 5 O 7 -3 Citrate ions are found in plants and animals. It is widely distributed in food and is a naturally occurring component of the diet. It is a common metabolic regulator of oxidative metabolism. It is a product of lactic acid bacteria and an important component of bone. Many citrates have dietary uses, including: It has been designated GRAS (Generally Regarded as Safe) by the U.S. Food and Drug Administration. There are. [Table 1]

[0064] For example, citric acid, citric acid monohydrate, anhydrous citric acid, sodium citrate, anhydrous citric acid Various citric acid-containing compounds, including trisodium citric acid and trisodium citrate dihydrate, are also available. As an ingredient in parenteral drug products according to the U.S. Food and Drug Administration's Inactive Ingredients Database The particular citric acid compounds used in the compositions of the present invention may be in acidic form or in various forms. salt forms, in particular the alkali (e.g., sodium and potassium) salts thereof and / or It may be monohydrate or dihydrate. Of course, sodium citrate is preferred. In embodiments, the concentration of citrate in the compositions of the present invention is from about 10 to about 35 mM or In a preferred embodiment, the citrate concentration ranges from about 10 to about 35 mM. In certain preferred embodiments, the concentration ranges from about 15 mM to about 30 mM. The citrate concentration ranges from about 15 to about 25 mM. In certain embodiments, the citrate concentration In certain embodiments, the concentration of citrate ranges from about 15 to about 25 mM. 20 mM or about 20 mM to about 25 mM. Particular compositions include about 15, about 20, about 2 5, about 30 or about 35 mM citrate concentration.

[0065] Although the addition of citrate has been found to result in an improvement in time response, The addition of citrate also poses a greater disadvantage from a stability standpoint. In order to have sufficient chemical and physical stability for long-term storage and use, The composition may further comprise stabilizing agents such as zinc, magnesium, chloride, and surfactants. It contains or comprises an agent.

[0066] Zinc oxide may be added to provide the desired stoichiometry of zinc ions. It has two specific, high-affinity zinc-binding sites. The ion is sometimes called "bound" zinc. Currently available zinc-containing preparations include There are approximately 2-4 zinc ions per insulin hexamer. The insulin composition contains 6 molecules of insulin (HUMULIN® RU-500) It contains about 2.4 ions of zinc, some of which is equivalent to six molecules of insulin (HUMALOG 3.0 ions of zinc per unit of Insulin. Insulin aspart (HUMALOG®) and insulin aspart (NOVOLO®) The 100 U / mL formulation of insulin (G) contains approximately 3.0 mg of zinc per 6 molecules of insulin. ions, which corresponds to a concentration of about 0.3 mM. Currently available 200 U / mL formulations have approximately 3.5 in / mL of zinc per 6 molecules of insulin. on, which corresponds to a zinc concentration of about 0.7 mM. Eli Lilly and Human insulin sold by the Company (HUMULIN®) The currently available 100 U / mL formulation of insulin contains approximately 2.3 units of zinc per 6 molecules of insulin. ions, which corresponds to a zinc concentration of about 0.23 mM.

[0067] The compositions of the invention contain at least enough insulin molecules to form stable hexamers. The composition of the present invention has a zinc concentration sufficient to provide sufficient zinc ions. The supplement must contain sufficient zinc to provide at least two ions of zinc per insulin hexamer. Certain embodiments of the composition of the present invention contain about 0.2 mM to about 0.8 mM zinc. A specific embodiment of the present invention is when the insulin concentration is 100 IU / mL. The zinc concentration in insulin is about 0.3 mM (about 3.0 Zn / 6 insulin molecules). , about 100 U / mL, about 200 U / mL, about 300 U / mL, or about 500 U / mL In a particular embodiment of the invention having insulin concentrations of 2% zinc per insulin hexamer. The minimum zinc concentrations required to provide the ions were about 0.2 mM, about 0.4 mM, and about 0.5 mM, respectively. This would be about 0.6 mM or about 1 mM.

[0068] Excess zinc inclusion - i.e., more zinc than can be bound by insulin hexamers Such zinc may be present in the form of "free" or "unbound" zinc. In certain compositions of the present invention, excess free or unbound zinc is Inclusion of zinc in the diet has been found to have a stabilizing effect. Compositions having sulinspro and a zinc concentration of up to about 1 mM include those having about 0.2 mM bound zinc. and approximately 0.8 mM of unbound or free zinc - fast acting and stable However, too much free or unbound zinc has been found to be involved. Inclusion of insulin at about 100 U / mL may attenuate the improvement in time response. A composition having a zinc concentration of about 5 mM with spro constitutes about 4.8 mM of unbound zinc. - but without the improvement in time action seen with compositions with lower zinc concentrations. In a particular embodiment of the invention, the concentration of zinc is from about 0.2 to about 2 mM, about 0.4 to about 1 mM, or about 0.6 to about 0.9 mM. , the concentration of zinc is about 0.6, about 0.7, about 0.8 or about 0.9 mM.

[0069] The magnesium (Mg +2 ) also has a stabilizing effect. Magnesium ions can be obtained in a variety of ways, for example, by using MgCl 2 minutes This may be provided through the addition of magnesium chloride having the formula and a molecular weight of 95.211. stomach.

[0070] While magnesium may have a stabilizing effect in certain compositions, the concentration of citrate Concentrations above this level will result in insulin precipitation. The maximum amount of ammonium is limited by the amount of citrate included. MgCl as a stabilizer in 2 When used, the ratio of magnesium citrate to The molar ratio of magnesium chloride to citrate ranges from about 1:10 to about 1:2. The molar ratio of citrate to acetic acid ranges from about 1:5 to about 1:3. Thus, for example, Magnesium citrate in a ratio of about 1:10 to about 1:2 in a composition that is between 10 to about 30 mM To achieve this molar ratio of magnesium to salt, the concentration is between about 1 and about 15 mM. Similarly, in a composition having a citrate concentration of 25 mM, the magnesium The magnesium concentration ranges from 5 to 8.3 mM to achieve a molar ratio of magnesium to citrate. In certain embodiments, the magnesium concentration ranges from about 1 to about 15 mM. In embodiments, the magnesium concentration is about 1 to about 5 mM, about 5 to about 10 mM, or The concentration of magnesium chloride in certain embodiments of the present invention ranges from about 10 to about 15 mM. ranges from 1 to about 5 mM (about 0.48 mg / mL). In certain embodiments, magnesium The concentration is about 5 mM.

[0071] The chloride ions (Cl - The inclusion of a certain concentration of ) also has a stabilizing effect. Chloride ions have been found to provide magnesium as described above. MgCl 2 or through the addition of sodium chloride. Certain embodiments of the composition of the present invention include sodium chloride. The molecular formula of thorium is NaCl and its molecular weight is 58.44. For example, APIDRA® (insulin) containing 5 mg / mL sodium chloride inguridine, NOVOLOG® 0.58 mg / mL sodium chloride Some currently available preparations of rapid-acting insulin analogues, such as insulin aspart, It is used in medicines.

[0072] For example, MgCl 2 or the inclusion in the composition of a chloride-containing excipient such as NaCl. The addition of a certain amount of chloride via the cellulose acetate solution has been found to have a stabilizing effect, while If the total chloride content of the composition is too high, the insulin in the composition may crystallize at low temperatures. Thus, the total chloride content of the composition must be taken into consideration.

[0073] Magnesium chloride and / or chlorine as stabilizers to determine the total chloride content of the composition. The chloride ions, which can be added via the addition of sodium chloride, as well as the insulin ballast, Addition of small amounts of HCl that may be necessary for pH adjustment with the active pharmaceutical ingredient (API) A solution prepared by solubilization of zinc oxide (ZnO) through addition of HCl and / or by Chloride ions through the addition of other ingredients together with the provision of Zn, which may be added in liquid form. must also be taken into consideration.

[0074] In certain embodiments, sodium chloride is used to reach a target chloride concentration or concentration range. Provide the amount of additional chloride needed to make the solution—i.e., the amount of sodium chloride added. The amount of insulin can vary from the target chloride concentration to pH adjustment, for example, via the addition of insulin API. Magnesium chloride and / or H2O may be required for stabilization of zinc oxide and / or By subtracting the amount of chloride provided through the addition of other components, For example, if the target chloride concentration in the formulation is about 20 mM and M chloride is used in bulk insulin API, magnesium chloride and pH adjustment If the drug is provided via a combination of 5 mM sodium chloride, then 5 mM sodium chloride must be added. Those skilled in the art will appreciate that the amount of sodium chloride added to such formulations will depend on (1) other sources The additional chloride that would be required based on the theoretical chloride content added via the source (2) Prepare the formulation with all excipients except sodium chloride in advance. The chloride content of the formulation is then measured and compared with the target chloride concentration or concentration range. It will be appreciated that the amount of water present in the aqueous formulation may be determined by calculating the difference between the amount of water present in the aqueous formulation and the amount of water present in the aqueous formulation. The chloride content of the solution can be determined by various known techniques, such as, for example, titration or the ion-selective electrode method. It may be measured using

[0075] In addition, the cold crystallization challenges associated with relatively high chloride concentrations are sensitive to citrate concentrations. Thus, citric acid at the lower end of the ranges provided herein has been found to be Compositions of the invention having salt concentrations include citrate concentrations at the upper end of the ranges provided herein. For example, a composition having 25 m The addition of high concentrations of sodium chloride, such as 50-75 mM, to formulations containing 10 mM citrate It has been observed that low-temperature crystallization challenges can be achieved with 50 mM When sodium chloride is added to a 15 mM citrate formulation or a 25 mM sodium chloride formulation, This was not consistently observed when 25 mM citrate was added to the formulation.

[0076] For purposes of clarity, as used herein, the term "chloride" or "total chloride" refers to With respect to the addition of any ingredient, a composition that is stated to contain, for example, 25 mM chloride A source of chloride ions in the preparation, or required for pH adjustment or ZnO solubilization. MgCl 2 , including chloride provided via addition of NaCl and / or HCl Total chloride refers to the total amount of chloride ions in the composition provided. Magnesium chloride, {MgCl 2 ", "Sodium chloride" and "NaCl" are used in the composition The amounts of these particular salts added to the solution refer to the amount of magnesium chloride added to the solution. Thus, 5 mM magnesium chloride and 10 mM In a composition described as containing sodium chloride, for example, chloride or total Chloride concentrations include magnesium chloride, sodium chloride, and other salts such as HCl. The combined amount of chloride ions provided by other sources is included.

[0077] In certain embodiments of the present invention, the total chloride concentration ranges from about 10 to about 60 mM. In embodiments, the total chloride concentration ranges from about 15 to about 35 mM. The chloride concentration ranges from about 20 to about 25 mM. In certain embodiments, the total chloride concentration is about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM or about 25 mM.

[0078] In certain embodiments of the present invention, the composition comprises sodium chloride at a concentration ranging from about 1 to about 50 mM. In certain embodiments, the concentration of sodium chloride is about 1 to about 25 mM. In embodiments, the concentration of sodium chloride ranges from about 1 to about 20 mM. The concentration of sodium chloride in the form is about 15 mM (about 0.88 mg / mL) to about 25 mM M (about 2.0 mg / mL). In certain embodiments, the concentration of sodium chloride ranges from about 1 , about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13 , about 14, about 15, about 16, about 17, about 18, about 19 or about 20 mM.

[0079] Optionally, to achieve sufficient chemical and physical stability, the composition may further comprise one or more The composition may contain additional stabilizers as described above. Exemplary stabilizers include surfactants. In particular, the combination of a surfactant and magnesium as a stabilizer in the composition of the present invention It has been discovered that these compounds can have greater than additive or synergistic stabilizing effects.

[0080] Examples of surfactants disclosed for use in parenteral pharmaceutical compositions include polysorbates Polysorbates such as 20 (TWEEN® 20), PEG 400, PEG 3 000, polyethylene glycol such as TRITON (trademark) X-100, polyethylene (23) Lauryl ether (CAS number: 9002-92-0, product name BRIJ (registered polyethylene glycols, such as those sold under the trademark MYRJ Alkoxylated fatty acids such as polypropylene glycols, block copolymers, e.g. For example, poloxamer 188 (CAS number 9003-11-6, trade name PLURONIC (registered trademark) F-68) and Poloxamer 407 (PLURONIC® ) F127), sorbitan alkyl esters (e.g., SPAN®), poly Ethoxylated castor oil (e.g., KOLLIPHOR®, CREMOPHOR®) (registered trademark) and trehalose and its derivatives, such as trehalose laurate. In certain embodiments, the surfactant is a polyoxyethylene (2 3) Consists of lauryl ether, poloxamer 188 and trehalose laurate A preferred surfactant is poloxamer 188. For example, the concentration of a surfactant such as Poloxamer 188 ranges from about 0.001 to about 2% w / v, about 0.001 to about 0.2% w / v, about 0.03 to about 0.12% w / v, Or about 0.06 to about 0.09% w / v. In certain embodiments, poloxamer 1 The concentration of 88 is about 0.06, about 0.07, about 0.08 or about 0.09% w / v.

[0081] In addition, the compositions of the present invention may include one or more preservatives, which provide antimicrobial properties, and further The composition may provide a stability benefit when first manufactured. If the composition is provided in a vial or cartridge for use, compatibility with other ingredients of the formulation Antimicrobial preservative compounds or mixtures of compounds that are The strength of the additive is sufficient to meet the requirements of the US Pharmacopeia Mo nographs.Insulin lispro injection.USP29- NF24;British Pharmacopeia Monographs 200 8 Volume III:Insulin aspart injection;U. S.Pharmacopeia Monographs.Insulin assays ; and US Pharmacopeia general chapters.USP 29-NF24.Rockville,MD:USPharmacopeial C onvention;2005.Antimicrobial efficacy ss testing; see pp. 2499-2500. The preferred preservative is ant The fatty acids are selected from the group consisting of carboxylic acids and phenolic compounds, or mixtures of such compounds.

[0082] Thus, the compositions of the present invention contain one or more preservatives. The concentrations of preservatives commonly used in insulin products are , Phenol (CAS number 108-95-2, molecular formula C 6 H 5 OH, molecular weight 94.11, ), and m-cresol (CAS number 108-39-4, molecular formula C 7 H 8 O, molecular weight 10 8.14). Current commercially available compositions include, for example, 3.15 mg / mL m- Cresols (HUMALOG® and APIDRA®), 1.72 m g / mL m-cresol and 1.50 mg / mL phenol (NOVOLOG Trademark), and 2.5 mg / mL m-cresol (HUMULIN® The compositions of the present invention contain one or more preservatives. The agent is selected from the group consisting of phenol and meta-cresol and mixtures thereof. Alternatively, the preservative is meta-cresol. In certain embodiments, the concentration of meta-cresol is The concentration of meta-cresol is about 2.5 mg / mL to about 3.8 mg / mL. Preferably, the concentration of meta-cresol is Approximately 3.15 mg / mL.

[0083] Solutions that are not nearly isotonic with body fluids may cause a painful or stinging sensation when administered, so When administering the composition, it should be administered as close as possible to the isotonicity (i.e., osmotic pressure) of the body fluids at the injection site. It is therefore desirable for the composition to be approximately isotonic with the body fluids at the site of injection. It is preferable that the osmotic pressure of the composition in the absence of an isotonicity agent is less than the osmotic pressure of the tissue (30% for blood). 0 mOsmol / kg; the European Pharmacopoeia requirement for osmolality is >240 mOsmol / k g), then a tonicity agent is generally added to improve the isotonicity of the composition. The typical tonicity agent is glycerol. (glycerin) and sodium chloride. If the addition of an isotonicity agent is required, Glycerol is preferred. The amount of tonicity agent to be added is readily determined using standard techniques. Ru. Remington:The Science and Practice of Pharmacy,David B.Troy and Paul Beringer, eds., Lippincott Williams & Wilkins, 2006, pp.257-259;Remington:Essentials of Pharm aceutics,Linda Ed Felton,Pharmaceutical Press, 2013, pp. 277-300. In certain embodiments, the concentration of glycerol The concentration is about 1 to about 16 mg / mL.

[0084] In certain embodiments, the pharmaceutical compositions are stable under the conditions of storage and use. As used, the term "stable" refers to, for example, less than 5% loss of insulin potency, 1. Formation of A-21 desamido less than 5%, insulin and A-21 desamido less than 4% Occurrence of other substances (collectively called other related substances (ORS)), less than 1.5% macromolecule The chemistry as indicated by high molecular weight (HMW) formation and maintenance of a clear, colorless solution without precipitation. Such properties refer to both chemical and physical stability. In certain embodiments, the ion exchange rate may be measured by known techniques, including those summarized in The pharmaceutical composition is stable enough to allow storage at 2-8°C for at least 24 months. In an embodiment, the pharmaceutical composition is a vial or cartridge for a reusable pen injector. It is stable enough to allow up to 28 days of use at temperatures up to 30°C for cartridges. In this embodiment, the pharmaceutical composition has a shelf life sufficient to permit storage at 2-8° C. for at least 36 months. In certain embodiments, the pharmaceutical composition is a reusable pen injector. Allows up to 32 days of use at temperatures up to 30°C for vials or cartridges In certain embodiments, the composition is stable for up to 7 days of continuous subcutaneous insulin infusion therapy. The composition is stable enough to allow its use in pumping devices for

[0085] As mentioned above, citric acid is added to contribute to the improvement in time action. Salts are also known to have buffering properties at certain pH levels, but additional salts can be added if desired. Examples of such buffer compounds include, for example, sodium phosphate dibasic. Phosphate buffers such as sodium thorium, or sodium acetate and tris(hydroxymethyl) The preferred buffer is phosphate or TRIS. The pH of insulin compositions is usually in the range of 7.2 to 7.6, with typical target pH values ​​being The pH of the composition of the present invention is usually 7.0 to 7.8, which is It is prepared using a physiologically appropriate acid and base, usually 10% hydrochloric acid and 10% sodium hydroxide. Preferably, the pH is about 7.4.

[0086] The route of administration of the composition of the present invention may be intravenous, intradermal or intraperitoneal. However, it is usually administered subcutaneously, for example by self-administration with a syringe or pen. or by continuous subcutaneous insulin infusion therapy via an insulin pump device. Preferably, the route of administration is by self-administered subcutaneous injection. The dose of active agent injected is determined by the patient's The decision regarding whether or not to discontinue use of the treatment will be made by the patient in consultation with their doctor.

[0087] Additional embodiments of the invention include those described below. Contains insulin, citrate, zinc, magnesium, chloride, surfactants and preservatives Pharmaceutical composition.

[0088] Insulin may be human insulin, insulin lispro, insulin aspart or insulin The pharmaceutical composition of any of the preceding embodiments, wherein the compound is selected from the group consisting of slingular lysine.

[0089] The pharmaceutical composition of any of the preceding embodiments, wherein the insulin is insulin lispro.

[0090] The pharmaceutical composition of any of the above-mentioned embodiments, wherein the insulin concentration is about 100 to 500 U / mL. composition.

[0091] The pharmaceutical composition of any of the above-mentioned embodiments, wherein the insulin concentration is about 100 to 300 U / mL. composition.

[0092] The pharmaceutical composition of any of the above-mentioned embodiments, wherein the insulin concentration is about 100 to 200 U / mL. composition.

[0093] The pharmaceutical composition of any of the preceding embodiments, wherein the insulin concentration is about 100 U / mL. The pharmaceutical composition of any of the preceding embodiments, wherein the insulin concentration is about 100 U / mL.

[0094] The pharmaceutical composition of any of the above embodiments, wherein the concentration of citrate is about 10 to about 35 mM. thing.

[0095] The pharmaceutical composition of any of the above embodiments, wherein the concentration of citrate is about 10 to about 30 mM. thing.

[0096] The pharmaceutical composition of any of the preceding embodiments, wherein the concentration of citrate is about 15 to about 35 mM. thing.

[0097] The pharmaceutical composition of any of the above embodiments, wherein the concentration of citrate is about 15 to about 30 mM. thing.

[0098] The pharmaceutical composition of any of the above embodiments, wherein the concentration of citrate is about 15 to about 25 mM. thing.

[0099] The pharmaceutical composition of any of the preceding embodiments, wherein the concentration of citrate is about 15 to about 20 mM. thing.

[0100] The pharmaceutical composition of any of the above embodiments, wherein the concentration of citrate is about 20 to about 25 mM. thing.

[0101] The pharmaceutical composition of any of the preceding embodiments, wherein the concentration of zinc is about 0.2 to about 2 mM.

[0102] The pharmaceutical composition of any of the preceding embodiments, wherein the concentration of zinc is about 0.3 to about 1 mM.

[0103] The pharmaceutical composition of any of the preceding embodiments, wherein the concentration of zinc is about 0.4 to about 1 mM.

[0104] The pharmaceutical composition of any of the above embodiments, wherein the concentration of zinc is about 0.6 to about 0.9 mM. .

[0105] The molar ratio of magnesium to citrate is about 1:10 to about 1:2. 2. The pharmaceutical composition of any of the following forms:

[0106] The above embodiment, wherein the molar ratio of magnesium to citrate is about 1:5 to about 1:3. The pharmaceutical composition according to any one of claims 1 to 4,

[0107] The pharmaceutical composition of any of the above embodiments, wherein the concentration of magnesium is about 1 to about 15 mM. thing.

[0108] The pharmaceutical composition of any of the above embodiments, wherein the concentration of magnesium is about 1 to about 5 mM. .

[0109] The pharmaceutical composition of any of the above embodiments, wherein the concentration of magnesium is about 5 to about 10 mM. thing.

[0110] The pharmaceutical composition of any of the above-mentioned embodiments, wherein the concentration of magnesium is about 10 to about 15 mM. Composition.

[0111] The pharmaceutical composition of any of the above embodiments, wherein the concentration of magnesium is about 3 to about 8 mM. .

[0112] The above embodiment, wherein the concentration of magnesium is provided through the inclusion of magnesium chloride. Any pharmaceutical composition.

[0113] The pharmaceutical composition of any of the preceding embodiments, wherein the total chloride concentration is about 10 to about 60 mM. .

[0114] The pharmaceutical composition of any of the preceding embodiments, wherein the total chloride concentration is about 15 to about 35 mM. .

[0115] The pharmaceutical composition of any of the preceding embodiments, wherein the total chloride concentration is about 15 to about 30 mM. .

[0116] The pharmaceutical composition of any of the above embodiments, wherein the total chloride concentration is about 20 to about 25 mM. .

[0117] Any of the above embodiments, wherein the composition comprises sodium chloride at a concentration ranging from about 1 to about 50 mM. Any pharmaceutical composition.

[0118] Any of the above embodiments, wherein the composition comprises sodium chloride at a concentration ranging from about 1 to about 25 mM. Any pharmaceutical composition.

[0119] Any of the above embodiments, wherein the composition comprises sodium chloride at a concentration ranging from about 1 to about 20 mM. Any pharmaceutical composition.

[0120] Any of the above-mentioned embodiments, wherein the concentration of the surfactant is about 0.001 to about 0.2% w / v. A pharmaceutical composition.

[0121] The pharmaceutical composition of any of the preceding embodiments, wherein the surfactant is poloxamer 188.

[0122] The above embodiment, wherein the concentration of poloxamer 188 is about 0.003 to about 0.2% w / v. Any of the pharmaceutical compositions.

[0123] The above embodiment, wherein the concentration of poloxamer 188 is about 0.03 to about 0.12% w / v. Any of the pharmaceutical compositions.

[0124] The above embodiment, wherein the concentration of poloxamer 188 is about 0.06 to about 0.09% w / v. Any of the pharmaceutical compositions.

[0125] The method of any one of the preceding embodiments, wherein the preservative is selected from the group consisting of phenol and meta-cresol. Any pharmaceutical composition.

[0126] The pharmaceutical composition of any of the previous embodiments, wherein the preservative is meta-cresol.

[0127] Any of the above embodiments, wherein the concentration of meta-cresol is about 2.5 to about 3.8 mg / mL. Any of the pharmaceutical compositions.

[0128] The pharmaceutical composition of any of the preceding embodiments, further comprising a tonicity agent.

[0129] The pharmaceutical composition of any of the previous embodiments, comprising glycerol as a tonicity agent.

[0130] The above embodiment includes glycerol as a tonicity agent at a concentration of about 1 to about 16 mg / mL. Any of the pharmaceutical compositions.

[0131] The pharmaceutical composition of any of the preceding embodiments, further comprising an additional buffer.

[0132] The pharmaceutical composition of any of the preceding embodiments, wherein the pH of the composition is about 7.0 to about 7.8.

[0133] The composition is less effective than a composition containing the same insulin but without citrate. Any of the above embodiments provides insulin uptake into the blood that is 20% more rapid. Any of the pharmaceutical compositions.

[0134] The composition is less effective than a composition containing the same insulin but without citrate. The pharmaceutical composition of any of the preceding embodiments, providing an onset of action that is 20% more rapid than either of the preceding embodiments.

[0135] The composition is less effective than a composition containing the same insulin but without citrate. Any of the above embodiments provides insulin uptake into the blood that is 30% more rapid. Any of the pharmaceutical compositions.

[0136] The composition is less effective than a composition containing the same insulin but without citrate. The pharmaceutical composition of any of the preceding embodiments, providing an onset of action that is 30% more rapid than either of the preceding embodiments.

[0137] The composition is stable enough to allow storage at 2-8°C for at least 24 months. The pharmaceutical composition of any of the embodiments.

[0138] The composition is suitable for use in reusable pen-type injector vials or cartridges at 30°C. Any of the above embodiments is stable enough to allow up to 28 days in use at temperatures up to 280°C. A pharmaceutical composition.

[0139] The composition is stable enough to allow storage at 2-8°C for at least 36 months. The pharmaceutical composition of any of the embodiments.

[0140] The composition is suitable for use in reusable pen-type injector vials or cartridges at 30°C. Any of the above embodiments is stable enough to allow up to 32 days in use at temperatures up to 400°C. A pharmaceutical composition.

[0141] The composition allows for use in a pump device for continuous subcutaneous insulin infusion therapy for up to 7 days. The pharmaceutical composition of any of the preceding embodiments, wherein the pharmaceutical composition is stable to

[0142] The pharmaceutical composition of any of the preceding embodiments, wherein the composition does not include an additional chelating agent.

[0143] The pharmaceutical composition of any of the preceding embodiments, wherein the composition does not contain EDTA.

[0144] The composition is EDTA, ethylene glycol tetraacetic acid (EGTA), alginic acid, alpha Lipoic acid, dimercaptosuccinic acid (DMSA), or 1,2-diaminocyclohexane tetrahydrofuran The pharmaceutical composition of any of the preceding embodiments, which does not contain acetic acid (CDTA).

[0145] The pharmaceutical composition of any of the preceding embodiments, wherein the composition does not include any vasodilators.

[0146] The pharmaceutical composition of any of the above embodiments, wherein the composition does not include any nitroglycerin. .

[0147] The composition comprises adenosine, endothelium-derived hyperpolarizing factor, and phosphodiesterase type 5 (PDES). Inhibitors, potassium channel openers, prostacyclin, forskolin, nitroglycerin The above-mentioned embodiment does not contain serine, nitric oxide formers, amyl nitrite, or nitroprusside. 2. The pharmaceutical composition of any of the following forms:

[0148] The pharmaceutical composition of any of the preceding embodiments, wherein the composition does not comprise any oligosaccharides.

[0149] Administering an effective dose of the pharmaceutical composition of any of the above embodiments to a human in need thereof. 46. ​​A method for treating diabetes comprising:

[0150] The pharmaceutical composition of any of the above embodiments for use in a method of treatment.

[0151] The pharmaceutical composition of any of the above embodiments for use in treating diabetes.

[0152] Any of the above embodiments for use in the manufacture of a medicament for the treatment of diabetes. Pharmaceutical compositions.

[0153] A multi-use vial comprising any one of the pharmaceutical compositions of any of the above embodiments.

[0154] A reusable injection pen comprising any one of the pharmaceutical compositions of any of the above embodiments. vessel.

[0155] Continuous subcutaneous insulin infusion comprising any one of the pharmaceutical compositions of any of the above embodiments. Pumping equipment for therapy.

[0156] This invention is further illustrated by the following examples, which should not be construed as limiting. It's not good. EXAMPLES

[0157] Pharmacokinetic and Pharmacodynamic Studies 35 mM citrate, 0.3 mM zinc, and 5 mM MgCl 2 and 23 mM NaCl Coformulated insulin lispro A diabetic (alloxan-induced) castrated male with a previously secured vascular access port Yucatan miniature pigs were used under the supervision of staff and veterinarians. Diabetic animals were kept individually They were kept in a cage and allowed free access to fresh water. They were fed a standard diet twice daily and an appropriate diet twice daily. Maintenance baseline and prandial insulin is given to manage the diabetic condition.

[0158] The test article (Composition A in the table below) was formulated and sent to the testing site. The og® insulin control was a commercially available vial available locally at the study site. Table 1 [Table 2]

[0159] On the day before the study, animals were fed half the daily allotment of chow and given a morning maintenance dose of 0.2 U / kg of Humalog® Mix 75 / 25 insulin. All test animals The mice were fasted overnight and were deprived of evening insulin or food prior to drug administration on the test day. Ta.

[0160] On the morning of the study, all animals were placed in a sling for restraint and the vascular access port (for blood sampling) was inserted. The endoscopic catheter (provided for this purpose) was accessed and checked for patency. Animals were randomly assigned to treatment groups. This was a complete crossover design with n=13. One animal participated in the Composition A treatment group. No additional treatment was added, so n=12 in that treatment group.

[0161] Two baseline blood samples (-30 and -20 min) were taken, after which the animals were returned to their cages. The rats were fed approximately 300 g of food. 20 min after presentation of complete food consumption, the rats were given a Terumo insulin injection. The test article was administered subcutaneously into the animal's flank using a syringe (0.3 or 0.5 mL with a 1 / 2" needle). All test animals had access to clean, fresh water throughout the remaining blood collection period. Seth.

[0162] The following time points: −30, −20 (followed immediately by food), 0 (just before dosing), s.c. After 5, 10, 15, 30, 45, 60, 75, 90, 105, 120, 150, 180, Serial blood samples (2.0 mL each) were taken from each animal at 240 and 360 min.

[0163] Blood samples (anticoagulant: none [serum]) were incubated for at least 30 minutes, but not more than 2 hours. The serum was then separated by centrifugation and divided into two aliquots. The mixture was divided into 10 portions and stored frozen at approximately -70°C.

[0164] Serum glucose concentrations were measured using an automated AU480 clinical laboratory analyzer (Beckman Coult The measurements were performed using a PK aliquot. The package will be shipped via overnight delivery service on dry ice to EMD Millipore Cor , St. Charles, MO, and included a detailed sample receipt.

[0165] Serum glucose data are presented as mean (mg / dL) ± SEM unless otherwise specified. As shown in Table 2 below. Table 2 [Table 3]

[0166] Microanalysis was performed to statistically compare serum glucose changes at different time points and under different formulations. The change in blood glucose at each time point was calculated as the first derivative (instantaneous differential) at the current time point. Polynomials of degree 10 or less are fitted to the time course data for each individual animal. The Bayesian Information Criterion (BIC) is used to select the optimal order of the polynomial. The rate of change in the curve is calculated as the derivative of the fitted polynomial curve at that point. Once the derivatives were obtained, an ANOVA model was fitted to compare the different formulations at each time point. Animal-to-animal variability is accounted for in the ANOVA model. Contrasts are constructed to compare the different formulations. The p-values ​​are adjusted using the R package “multcomp.” For time points beyond 150 min, the derivatives are As there were not enough time points to make a robust estimate, these time points were excluded from the statistical analysis.

[0167] Composition A was significantly lower than Humalog® at 5, 10, and 15 minutes after administration. (p≦0.01) resulted in a faster decrease in serum glucose.

[0168] Using the total insulin RIA, the Composition A and Humalog® treated groups Insulin levels were measured in serum PK samples for each of the following: The upper and lower limits of quantification are 20 pM and 5000 pM, respectively. The value below the lower limit of quantification is 20 pM. It is considered that the non-compute Conduct treatment pharmacokinetic analysis. Table 3 [Table 4] Abbreviations: Tmax = time to maximum concentration, Cmax = maximum concentration, AUCINF = 0 to infinity Area under the curve up to 100 mg / kg, CL / F = clearance / bioavailability The mean and median Tmax were 55% and 55% lower, respectively, for Composition A than for Humalog. and 72% faster.

[0169] 25-35 mM citrate, 0.285 mM zinc, and 5 mM MgCl 2 and 15 to 23 Insulin Lispro formulated with 1 mM NaCl

[0170] Generally, diabetic (alloxan-induced) castrated male Yucatan militants were treated according to the procedure described above. Various concentrations of citrate, zinc, magnesium chloride and sodium chloride were added to the nibachi. The composition comprising the compound is tested.

[0171] Test articles (compositions A' and B in the table below) are formulated and shipped to the testing site and The alog® insulin control is a commercially available vial on-site at the study site. Table 4 [Table 5]

[0172] The study was a complete crossover study of 21 pigs, with all pigs examined across all treatments. Designed as planned (n=21). One animal had a baseline blood glucose of <200 mg / dl. One animal was removed from the Humalog® treatment group due to a non-patented port As a result, the Humalog treatment group was excluded, so n=19 in that treatment group. One animal was removed from the Composition A' treatment group due to a misreporting of the dose at injection. Therefore, n=20 in that treatment group. One animal had a baseline of <200 mg / dl. Glycemic control excluded Composition B treatment group, so n=20 in that treatment group.

[0173] Serum glucose data are presented in Table 5 below as mean (mg / dL) ± SEM. can be. Table 5 [Table 6]

[0174] To statistically compare the changes in serum glucose at different time points and under different formulations, Generally, differential analysis was performed according to the procedure described above. Compositions A' and B were each Significantly lower serum glucose (p<0.01) was observed at 5, 10, and 15 minutes after administration compared to log ≦0.01) rapid decline occurred.

[0175] Serum insulin concentrations and PK parameters were generated and analyzed generally as described above, and PK parameters were analyzed. The results are provided in Table 6 below. Table 6 [Table 7] The median Tmax was 33% higher for compositions A' and B than for Humalog®. and 67% faster, and the mean Tmax was 48% faster for composition B than for Humalog.

[0176] 15 mM citrate, 0.285 mM zinc, and 5 mM MgCl 2 and 15 mM NaCl Insulin lispro formulated with l

[0177] Generally, diabetic (alloxan-induced) castrated male Yucatan militants were treated according to the procedure described above. In the nibachi, a composition containing citrate, zinc, magnesium chloride and sodium chloride was tested. Composition C will be formulated and shipped to the testing site, and the Humalog® control will be This is a vial for sale. Table 7 [Table 8]

[0178] The study was a complete crossover study of 13 pigs, with all pigs tested across all treatments. Designed as planned (n=13). One animal had a baseline blood glucose of <200 mg / dl. One animal was removed from the Composition C treatment group due to a misreported dose at injection. Composition C was excluded from the treatment group, resulting in n=11 in that treatment group.

[0179] Serum glucose data are presented in Table 8 below as mean (mg / dL) ± SEM. do. Table 8 [Table 9]

[0180] Composition C improved glucose proteolysis compared to the Humalog® control profile. A shift occurred in the file.

[0181] Serum insulin concentrations and PK parameters were generated and analyzed generally as described above, and PK parameters were analyzed. The results are provided in Table 9 below. Table 9 [Table 10] The mean and median Tmax were 28% and 29%, respectively, lower for Composition C than for Humalog. 25% faster.

[0182] Formulated with 25 mM citrate, 0.285 mM zinc, and 25 mM NaCl. Insulin lispro

[0183] Generally, diabetic (alloxan-induced) castrated male Yucatan militants were treated according to the procedure described above. A test was performed on a composition containing 25 mM citrate, zinc, and sodium chloride in nibachi. cormorant.

[0184] The test article (composition D in the table below) was formulated and shipped to the testing site and administered to Humalog. The ® Insulin control was a commercially available vial available on-site at the study site. Table 10 [Table 11]

[0185] The study included three doses (0. The study consisted of composition D at 1, 0.2 and 0.4 units / kg. All pigs were treated. The study was designed as a complete crossover design with 21 pigs (n=21) to be studied. For the 0.4 unit / kg dose, two animals were excluded from the Composition D treatment group (one 1 due to disease and 1 due to port nonpatency, so n = 19 in that treatment group. One animal was excluded from the Humalog® 0.4 units / kg treatment group ( Because of port non-patentability, n=20 in that treatment group. All other doses were The result was n=21.

[0186] Serum glucose data are presented in Table 11 as mean (mg / dL)±SEM. Table 11 [Table 12]

[0187] To statistically compare the changes in serum glucose at different time points and under different formulations, Generally, derivative analysis was used according to the procedure described above. When compared to the 3 doses of Composition D (0.1, 0.2 and 0.4 U / mL) kg) all showed significantly (p≦0.01) faster blood glucose levels at 5, 10, and 15 minutes. This results in a decrease.

[0188] For the pharmacokinetic analysis, 11 pigs were randomized using an online randomization tool. The insulin levels for serum PK samples for these pigs were selected as total insulin. The assay is measured using a thoracic RIA. The lower and upper limits of quantification for the assay are 20 Values ​​below the lower limit of quantification are considered to be 20 pM. Non-compartmental pharmacokinetic analysis using hoenix WinNonlin v6.3 Do the following. Table 12 [Table 13] The median Tmax value was 0.1 U / mL compared to the same dose with Humalog®. In the composition D group, the rate was 67%, 50%, and 40% at 0.2 U / kg, 0.2 U / kg, and 0.4 U / kg. The mean Tmax value was 0.1 times higher than the same dose with Humalog®. The total exposure and C max was higher than Humalog® at doses of 0.2U / kg and 0.4U / kg. It is also thought that the difference is larger for composition D.

[0189] 25 mM citrate and 5 mM MgCl 2 Human insulin, formulated with insulin Insulin aspart and insulin glulisine

[0190] Generally, diabetic (alloxan-induced) castrated male Yucatan militants were treated according to the procedure described above. The nibachi contained three different insulins, 25 mM citrate and magnesium chloride. The test is performed on a composition containing the above.

[0191] HUMULIN-R®, NOVOLOG® and APIDRA® Add enough citrate and magnesium chloride to a commercially available vial of 100 mM EDTA (100 mM NaCl) to make a 25 mM solution. The test article (Table 1 below) was added to the citrate and magnesium chloride to reach a concentration of 5 mM. Compositions E, F and G) are formulated. Table 13 [Table 14]

[0192] The study was designed as a complete crossover design with all pigs examined in all treatments. One animal had baseline blood glucose of <200 mg / dl and therefore did not tolerate the composition. F treatment group, resulting in n=19 in that group. Data are presented in Table 14 as mean (mg / dL) ± SEM.

[0193] Table 14 [Table 15]

[0194] Insulin aspart and citrate-containing formulations of human insulin are available from Novo Glucose profiles compared to log® and Humulin-R® controls Citrate-containing formulations of insulin glulisine produce a shift in the API profile. Less pronounced in the first approximately 90 minutes after administration compared to the dra® control A shift occurs.

[0195] Various concentrations of citrate and MgCl 2 and formulated with zinc, surfactants, and NaCl. Insulin lispro

[0196] Generally, diabetic (alloxan-induced) castrated male Yucatan militants were treated according to the procedure described above. In the nibita, various concentrations of citrate, magnesium chloride, zinc, surfactant and / or is tested on a composition containing sodium chloride.

[0197] Test compositions H and J in the following table are the active pharmaceutical ingredient ( The test articles I and K in the following table were prepared from Humalog® ) are prepared by adding citrate and the indicated stabilizers to vials of the formulation, The Humalog® insulin control was delivered to the study site on-site. This is a commercially available vial. Table 15 [Table 16]

[0198] The study was designed as a complete crossover design with all pigs examined in all treatments. One animal was not administered Composition I, J or any of the following due to port failure or the animal's condition: and K treatment groups, resulting in n = 18 for those treatment groups. Serum glucose data are presented in Table 16 as mean (mg / dL) ± SEM. .

[0199] Table 16 [Table 17]

[0200] The citrate-containing formulations improved glucose profiles compared to the Humalog® control. This causes a shift in the file.

[0201] Serum insulin concentrations and The PK parameters were generated and generally analyzed as described above, and the PK results are presented in Table 17 below. provide. Table 17 [Table 18]

[0202] The results show that compositions H, J and K have a faster PK onset than Humalog®. This shows that it has

[0203] Stability testing Insulin formulated with various concentrations of citrate, sodium chloride, and magnesium chloride Linris Pro

[0204] Accelerated shelf-life stability studies were performed to evaluate the To evaluate the stability of insulin lispro when co-formulated with insulin lispro and optionally magnesium chloride Stability samples having the compositions set forth in the following tables were prepared using the same formulations as the other excipients shown in the following tables. By formulating the active pharmaceutical ingredient insulin lispro together with and N), or Humalog® drug product vials containing citrate and sodium chloride Composition O is prepared by adding Table 18 [Table 19]

[0205] 0.22 micron PVDF membrane (Cat. No. SLGV013SL, Millex, Mi The solution was filtered using a 30 mL alipore filter with a screw top cap (Cat. No. C4015- 67A,National Scientific,Thermo Scientifi Glass vials (Cat. No. NC4015-1, National S Thermo Scientific, Inc. Incubate at 2-8°C and 30°C, respectively. Samples were taken at baseline, 8, 16, 27, 36, 44, 54, and 66 days. The incubation temperature for the analysis was 5°C from the start, 27, 36, 44, 54 and 66 Samples are removed at the day time points.

[0206] Size-exclusion high performance liquid chromatography (SEC-HPLC) analysis was performed to identify the protein Evaluate the efficacy of each composition at the stability time points using a UV detector at 276 nm. Quantify high molecular weight species. Isocratic elution of the mobile phase (0.1% TF) over a 25 min flow time. A, 50% ACN) at a flow rate of 1.0 mL / min on Sepax Zenix-C SEC- 80, 7.8 x 300 mm column of 3 μm particles (catalog number 233080-7 Separate each sample (10 μL) at room temperature using a 500 mL aliquot of insulin. The peak areas of insulin and insulin were compared by reversed-phase HPLC. The results reported in IU / mL are given below: show. Table 19 [Table 20] Table 20 [Table 21]

[0207] The insulin loss in the citrate-containing samples and the Humalog® control samples was It is less than 5% for all samples up to 66 days at 4°C and 30°C.

[0208] The percentage of high molecular weight (%HMW) is the total area percent of all peaks eluting prior to the main peak. The results (%HMW) are shown in the table below. Table 21 [Table 22] Table 22 [Table 23]

[0209] HMW formation was observed in all citrate-containing samples and the Humalog® control sample. The change was less than 1% up to 66 days at 4°C and 30°C for all samples.

[0210] Reverse-phase high-performance liquid chromatography (RP-HPLC) analysis was performed using UV at 214 nm. The detector is used to assess the purity of the protein in each composition at the stability time points. (50 mM sulfate, pH 2.3 + 20% acetonitrile (v / v)) and mobile phase B (5 0 mM sulfate, pH 2.3 + 50% acetonitrile (v / v)) at 0.6 mL / min Waters BioSuite C18 PA-B, 3.5 μm, 2.1× at a flow rate of 150 mm HPLC column (Part No. 186002435) or comparable Each sample (5 μL) was separated at 40°C using a column. The gradient of mobile phase B at 26, 27, 27.5 and 35.0 min was 21, 25, 25 , 30, 80, 80, 21 and 21%. Outside the main peak and the A-21 desamide The ratio of the sample to be analyzed is calculated by subtracting the ratio of the main peak and the ratio of A-21 desamido from 100 percent. Collectively, these peaks are associated with other related substances (ORS). The results (% outside the main peak and A-21 desamide) are shown below. Table 23 [Table 24] Table 24 [Table 25]

[0211] O for all citrate-containing samples and the Humalog® control sample RS was less than 1.5% after 66 days at 4°C and less than 3.0% after 66 days at 30°C. do. 25 mM citrate, 0.3 mM zinc, and 5 mM MgCl 2 and 0.09% poloxamer Physical stability of insulin lispro formulated with insulin and various concentrations of chloride.

[0212] 200 U / mL insulin lispros / mL and 32 mg insulin / mL in water for bulk sterile processing Lisp containing 6.30 mg of lysine / mL, 6.30 mg of metacresol / mL, and 0.6 mM zinc Prepare a concentrated stock preparation of the above. The pH of the preparation is adjusted with hydrochloric acid, and insulin lispro is added. Dissolve and then adjust to a pH of 7.45 with sodium hydroxide.

[0213] The insulin lispro concentrate was then dissolved in sodium citrate buffer, magnesium chloride 6 Hydrate powder, Poloxamer 188 solution, and granular sodium chloride diluted bulk sterile 23.2 mM (Composition P) or 17.7 mM (Composition Q) total salts in water at the appropriate volume for operation 25 mM citrate, 5 mM MgCl 2 , 0.09% poloxamer 1 88, 100 units / mL lispro, 16 mg / mL glycerin, 3.15 mg / mL A test formulation is prepared by making a composition of metacresol, and 0.3 mM zinc. .

[0214] The formulation is sterile filtered and then volumetrically transferred into 10 mL glass vials and filled to 7 mL. Then, plug and crimp seal.

[0215] Accelerated stability testing (30°C, shaking at 75 strokes per minute (spm)) was performed for 13 days. The vials of )) are visually inspected on days 2, 4, 8 and 13. They are clear and colorless with no precipitate. The vials containing the solution were considered scored as "passed," while the particulate matter Vials containing solutions that are not clear and / or discolored are classified as “incorrect.” The data is provided below in Table 25. Table 25 [Table 26]

[0216] 25 mM citrate and 0.3 mM zinc and 0.0 mM Zn with and without magnesium Physics of insulin lispro formulated with 9% poloxamer and 23 mM total chloride Stability

[0217] 200 U / mL insulin lispros / mL and 32 mg insulin / mL in water for bulk sterile processing Lisp containing 6.30 mg of lysine / mL, 6.30 mg of metacresol / mL, and 0.6 mM zinc Prepare a concentrated stock preparation of the above. The pH of the preparation is adjusted with hydrochloric acid, and insulin lispro is added. Dissolve and then adjust to a pH of 7.45 with sodium hydroxide.

[0218] The insulin lispro concentrate was then dissolved in sodium citrate buffer, poloxamer 188 Solution, granular sodium chloride, optionally diluted with magnesium chloride hexahydrate powder, bulk-free 25 mM citrate, 0.09% poloxamer 188, 10% ethanol in the appropriate amount of water for bacterial processing. 0 units / mL lispro, 16 mg / mL glycerin, 3.15 mg / mL metacrezo 0.3 mM zinc, 23 mM total chloride, and 0 mM (Composition R) or 5 mM (Composition Composition S) MgCl 2 A test formulation is prepared by making the composition as follows:

[0219] The formulation is sterile filtered and then volumetrically transferred into 10 mL glass vials and filled to 7 mL. Then, plug and crimp seal.

[0220] Generally, the above methods are followed by a 13-day accelerated stability test with periodic visual inspection. The data are provided below in Table 26. Table 26 [Table 27]

[0221] The results support that magnesium improved the stability of composition S compared to composition R. There are. 25 mM citrate and 5 mM MgCl 2 and 20 mM chloride, 0.6 mM zinc, and 0 Physical stability of insulin lispro formulated with .09% poloxamer.

[0222] 100 U / mL insulin lispro, 5 mM MgCl 2 , 20 mM chloride, 0. 6 mM zinc, 0.09% w / v poloxamer 188, 5 mg / mL glycerin, and and 3.15 mg / mL meta-cresol (Composition T), which has an accelerated storage period of 69 days. Conduct an interim test.

[0223] By lowering the pH by adding HCl, glycerin, metacresol, and lispro and zinc oxide in water for bulk sterile processing to prepare a bulk concentrate formulation. After adjusting the pH to 7.4-7.5 with NaOH, the final target excipient concentration of Composition T was achieved. Add a solution of citrate and additional stabilizers at the appropriate concentration to form a ballast. Sodium citrate buffer, zinc chloride and magnesium chloride in water for aseptic processing It is composed of poloxamer 188 and tetrahydrofuran. The pH can be adjusted to 7.4-7.5 as necessary. Afterwards, sodium chloride solution is added to achieve the target total chloride concentration. Bring formulation to volume with working water.

[0224] The formulation is sterile filtered and then volumetrically transferred to a 10 mL glass vial. Fill the vials to 100 mL, stopper and crimp seal. Store the vials stationary and upright at 30°C. Visual inspection is generally performed at days 36 and 69 as described above. All of the samples passed the test (n=3 at 36 days, n=5 at 69 days). Supported by the fact that product T remains clear and colorless without precipitate during accelerated shelf life testing. There are.

[0225] Various concentrations of citrate, magnesium, chloride, zinc, surfactant, glycerin, and T Insulin lispro formulated with RIS

[0226] The physical stability of the following seven formulations is examined in a 32-day simulated patient use study: Table 27 [Table 28]

[0227] Adding HCl to lower the pH will allow the appropriate amount of glycerol to be added to water for bulk aseptic processing. Concentrated by dissolving phosphorus, metacresol, lispro, zinc oxide and optionally TRIS. Concentrated lispro stock solutions are prepared. The compositions of the two concentrated solutions are as follows: (1) 200 U / mL Lispro, 12 mM total chloride, 10 mg / mL glycerin, 6.3 mg / mL metacresol, 0.6 mM total zinc, and (2) 200 U / mL Lisp B, 51 mM total chloride, 2.6 mg / mL glycerin, 6.3 mg / mL methacrylamide sol, 0.6 mM total zinc, 80 mM TRIS. Then, Adjust the solution to a pH of 7.4-7.5 using 100% sodium dodecyl sulfate and make up the volume with bulk sterile water. .

[0228] Sodium citrate buffer, zinc HCl solution, sodium hydroxide to adjust pH , magnesium chloride hexahydrate solution, poloxamer 188 solution, and sodium chloride solution Prepare the test composition by diluting the Lispro stock solution by sequentially adding The composition is sterile filtered and then volumetrically measured and made up to volume with water for bulk aseptic processing. The composition is volumetrically transferred to a 10 mL glass vial and filled to 10.3 mL. Then plug and crimp seal.

[0229] The compositions were stored at 30° C. and maintained under simulated in-use dosing conditions as described in the following steps. Obtain an insulin syringe and draw in 8 units of air; Insert the needle into the vial while in an upright position; make sure the tip of the needle touches the insulin solution. inject air into the vial; while holding the needle in the vial, Turn the syringe over; withdraw 8 units of the composition; if there are any air bubbles in the syringe, Gently move the plunger to push the air bubbles back into the vial; note that the final dose is 8 units. Adjust the syringe plunger; remove the syringe and place the vial in a 30°C incubator. Return to.

[0230] Visual inspection is generally performed at 15, 20, 25, 28 and 32 days according to the method described above. Data is provided in Table 28. Table 28 [Table 29]

[0231] In summary, the above tests were carried out using specific concentrations of citric acid and, for example, zinc, magnesium, The composition of insulin with other excipients such as sodium, chloride and surfactants is and has a faster glucose lowering effect, in certain embodiments It has been demonstrated that it is chemically and physically stable.

[0232] array Human insulin A chain GlyIleValGluGlnCysCysThrSerIleCysSerLeuT yrGlnLeuGluAsnTyrCysAsn (SEQ ID NO: 1) Human insulin B chain PheValAsnGlnHisLeuCysGlySerHisLeuValGluA laLeuTyrLeuValCysGlyGluArgGlyPhePheTyrTh rProLysThr (SEQ ID NO:2). Insulin lispro B chain PheValAsnGlnHisLeuCysGlySerHisLeuValGluA laLeuTyrLeuValCysGlyGluArgGlyPhePheTyrTh rLysProThr (SEQ ID NO: 3) Insulin aspart B chain PheValAsnGlnHisLeuCysGlySerHisLeuValGluA laLeuTyrLeuValCysGlyGluArgGlyPhePheTyrTh rAspLysThr (SEQ ID NO: 4) Insulin glulisine B chain PheValLysGlnHisLeuCysGlySerHisLeuValGluA laLeuTyrLeuValCysGlyGluArgGlyPhePheTyrTh rProGluThr (SEQ ID NO:5)

Claims

1. 1. A pharmaceutical composition comprising: (a) insulin; (b) citrate at a concentration of about 10 to about 30 mM; (c) zinc at a concentration of about 0.2 to about 2 mM; (d) magnesium at a concentration of about 1 to about 15 mM; (e) total chloride at a concentration of about 10 to about 60 mM; (f) a surfactant at a concentration of about 0.001 to about 0.2% w / v; (g) a preservative.

2. 2. The pharmaceutical composition of claim 1, wherein the insulin is insulin lispro.

3. 3. The insulin lispro according to claim 1, wherein the concentration of said insulin lispro is about 100 to about 500 U / mL.

2. The pharmaceutical composition according to claim 1 ,

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the concentration of the citrate is about 15 to about 25 mM. Pharmaceutical composition.

5. The pharmaceutical composition according to any one of claims 1 to 4, wherein the concentration of zinc is about 0.3 to about 1.1 mM. composition.

6. 6. The method of claim 1, wherein the magnesium concentration is provided through the inclusion of magnesium chloride. Any of the pharmaceutical compositions described above.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the magnesium concentration is about 3 to about 8 mM. composition.

8. The pharmaceutical composition according to any one of claims 1 to 7, wherein the concentration of the chloride is about 15 to about 35 mM. Composition.

9. 9. The pharmaceutical composition according to claim 1, wherein the concentration of the chloride is about 20 to about 25 mM. Composition.

10. 10. The method according to claim 1, further comprising administering to said patient a solution containing sodium chloride at a concentration ranging from about 1 to about 50 mM. A pharmaceutical composition comprising:

11. 11. The method according to claim 1, further comprising the step of: The pharmaceutical composition described above.

12. 12. The method according to claim 1, further comprising the step of: The pharmaceutical composition described above.

13. The pharmaceutical composition according to any one of claims 1 to 12, wherein the surfactant is poloxamer 188. Composition.

14. 14. The method according to claim 13, wherein the concentration of the poloxamer 188 is about 0.003 to about 0.2% w / v. The pharmaceutical composition described.

15. 15. The method according to claim 14, wherein the concentration of the poloxamer 188 is about 0.03 to about 0.12% w / v. The pharmaceutical composition described.

16. 16. The method according to claim 15, wherein the concentration of the poloxamer 188 is about 0.06 to about 0.09% w / v. The pharmaceutical composition described.

17. The preservative is selected from the group consisting of phenol and meta-cresol and mixtures thereof. The pharmaceutical composition according to any one of claims 1 to 16.

18. The pharmaceutical composition according to any one of claims 1 to 17, wherein the preservative is meta-cresol.

19. 19. The method of claim 18, wherein the concentration of the meta-cresol is from about 2.5 to about 3.8 mg / mL. Pharmaceutical compositions.

20. The pharmaceutical composition according to any one of claims 1 to 19, further comprising an isotonicity agent.

21. 21. The pharmaceutical composition of claim 20, wherein the tonicity agent is glycerol.

22. The pharmaceutical composition according to any one of claims 1 to 21, further comprising an additional buffer.

23. The pharmaceutical composition according to any one of claims 1 to 22, wherein the pH of the composition is about 7.0 to about 7.

8. composition.

24. A pharmaceutical composition comprising: (a) insulin lispro at a concentration of about 100 to about 200 IU / mL; (b) citrate at a concentration of about 15 to about 25 mM; (c) magnesium chloride at a concentration of about 3 to about 8 mM; (d) zinc at a concentration of about 0.6 to about 0.9 mM; (e) total chloride at a concentration of about 20 to about 25 mM; (f) poloxamer 188 at a concentration of about 0.06 to about 0.09% w / v; (g) meta-cresol at a concentration of about 2.8 to about 3.5 mg / mL; The pharmaceutical composition, wherein the pH of the composition is about 7.0 to 7.

8.

25. A pharmaceutical composition comprising: (a) insulin lispro at a concentration of 100 IU / mL; (b) citrate at a concentration of about 25 mM; (c) zinc at a concentration of about 0.9 mM; (d) magnesium chloride at a concentration of about 5 mM; (e) total chloride at a concentration of about 20 to about 25 mM; (f) poloxamer 188 at a concentration of about 0.09% w / v; (g) meta-cresol at a concentration of about 3.15 mg / mL; The pharmaceutical composition, wherein the pH of the composition is about 7.

4.

26. A pharmaceutical composition comprising: (a) insulin lispro at a concentration of 200 IU / mL; (b) citrate at a concentration of about 15 to about 25 mM; (c) magnesium chloride at a concentration of about 3 to about 8 mM; (d) zinc at a concentration of about 0.6 to about 0.9 mM; (e) total chloride at a concentration of about 20 to about 25 mM; (f) poloxamer 188 at a concentration of about 0.06 to about 0.09% w / v; (g) meta-cresol at a concentration of about 2.8 to about 3.5 mg / mL; (h) The pharmaceutical composition, wherein the pH of the composition is about 7.0 to 7.

8.

27. A pharmaceutical composition comprising: (a) insulin at a concentration of about 100 to about 200 IU / mL; (b) citrate at a concentration of about 15 to about 35 mM; and (c) magnesium chloride at a concentration of up to about 5 mM; (d) zinc at a concentration of about 0.2 to about 0.8 mM; (e) sodium chloride at a concentration of about 15 to about 25 mM; (f) a surfactant; and (g) meta-cresol at a concentration of about 2.8 to about 3.5 mg / mL; (h) The pharmaceutical composition, wherein the pH of the composition is about 7.0 to 7.

8.

28. The composition is less effective than a composition containing the same insulin but without citrate.

28. Any of claims 1 to 27, which provides insulin uptake into the blood that is at least 20% more rapid. The pharmaceutical composition according to any one of the preceding claims.

29. The composition has a shelf life of at least 24 months at 2-8°C and up to 28 days at temperatures up to 30°C.

28. The pharmaceutical composition according to any one of claims 1 to 27, which is stable enough to permit its use in

30. The pharmaceutical composition according to any one of claims 1 to 27, wherein the composition does not contain EDTA.

31. A pharmaceutical composition according to any one of claims 1 to 27, wherein said composition does not contain any vasodilator. thing.

32. A pharmaceutical composition according to any one of claims 1 to 27, wherein said composition does not contain any oligosaccharides. 。

33. A pharmaceutical composition according to any one of claims 1 to 27 in an effective dose for a human in need thereof.

23. A method for treating diabetes comprising administering

34. A pharmaceutical composition according to any one of claims 1 to 27 for use in therapy.

35. A pharmaceutical composition according to any one of claims 1 to 27 for use in the treatment of diabetes.

36. A pharmaceutical agent according to any one of claims 1 to 27 in the manufacture of a medicament for the treatment of diabetes. Use of the composition.

37. An article of manufacture comprising any one of the pharmaceutical compositions of claims 1-27.

38. 38. The article of manufacture of claim 37, which is a multi-use vial.

39. 38. The article of manufacture of claim 37, which is a reusable pen injection device.

40. 38. The article of manufacture of claim 37 which is a pump device for continuous subcutaneous insulin infusion therapy.

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