FORM 2
THE PATENTS ACT, 1970
(39 OF 1970)
As amended by the Patents (Amendment) Act, 2005 5
&
THE PATENTS RULES, 2003
As amended by the Patents (Amendment) Rules, 2005
COMPLETE SPECIFICATION
(Section 10 and rule 13) 10
TITLE OF THE INVENTION
MEDICAL INFUSION PUMP SYSTEM FOR THE DELIVERY OF AN 15 INSULIN COMPOUND
APPLICANTS 20
Arecor Limited, a Great Britain company, having its address at Chesterford Research Park, Little Chesterford, Saffron Walden CB10 1XL, Great Britain
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PREAMBLE TO THE DESCRIPTION
The following specification particularly describes this invention and the manner in which it is to be performed: 40
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FIELD OF THE INVENTION
This invention relates inter alia to an injection pen system for the delivery of an insulin compound, particularly rapid acting aqueous liquid pharmaceutical compositions of insulin and insulin analogues. Such a system is suitable for the treatment of subjects suffering from diabetes mellitus, especially Type 1 diabetes 5 mellitus.
BACKGROUND OF THE INVENTION
Diabetes mellitus (“diabetes”) is a metabolic disorder associated with poor control of blood sugar levels leading to hypo or hyperglycaemia. Untreated diabetes 10 can lead to serious microvascular and macrovascular complications including coronary artery disease, peripheral artery disease, stroke, diabetic nephropathy, neuropathy and retinopathy. The two main types of diabetes are (i) Type 1 diabetes resulting from the pancreas not producing insulin for which the usual treatment is insulin replacement therapy and (ii) Type 2 diabetes where patients either produce 15 insufficient insulin or have insulin resistance and for which treatments include insulin sensitising agents (such as metformin or pioglitazone), traditional insulin secretagogues (such as sulfonylureas), SGLT2 inhibitors (such as dapagliflozin, canagliflozin and empagliflozin) which reduce glucose absorption in the kidneys and so promote glucose excretion, GLP-1 agonists (such as exenatide and dulaglutide) 20 which stimulate insulin release from pancreatic beta cells and DPPIV inhibitors (such as sitagliptin or vildagliptin) which inhibit breakdown of GLP-1 leading to increased insulin secretion. Patients with Type 2 diabetes may eventually require insulin replacement therapy.
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For patients requiring insulin replacement therapy, a range of therapeutic options are possible. The use of recombinant human insulin has in recent times been overtaken by use of insulin analogues which have modified properties, for example, are longer acting or faster acting than normal insulin. Thus, a common regimen for a patient involves receiving a long acting basal insulin supplemented by a rapid acting 30 insulin around mealtimes.
Insulin is a peptide hormone formed of two chains (A chain and B chain, respectively 21 and 30 amino acids in length) linked via disulfide bridges. Insulin normally exists at neutral pH in the form of a hexamer, each hexamer comprising 35 three dimers bound together by zinc ions. Histidine residues on the insulin are known to be involved in the interaction with the zinc ions. Insulin is stored in the body in the hexameric form but the monomer form is the active form. Traditionally, therapeutic compositions of insulin have also been formulated in hexameric form in the presence of zinc ions. Typically, there are approximately three zinc cations per one insulin 40
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hexamer. It has been appreciated that the hexameric form is absorbed from the injection site considerably more slowly than the monomeric and dimeric forms. Therefore, a faster onset of insulin action can be achieved if the hexameric form is destabilised allowing a more rapid dissociation of the zinc-bound hexamer into dimers and monomers in the subcutaneous space following injection. Three insulin 5 analogues have been genetically engineered with this principle in mind. A first is insulin lispro (HUMALOG®) in which residues 28 and 29 of the B chain (Pro and Lys respectively) are reversed, a second is insulin aspart (NOVORAPID®) in which residue 28 of the B chain, normally Pro, is replaced by Asp, and a third is insulin glulisine (APIDRA®) in which residue 3 of the B chain, normally Asn is replaced by 10 Lys and residue 29 of the B chain, normally Lys, is replaced by Glu.
Whilst the existing rapid acting insulin analogues can achieve a more rapid onset of action, it has been appreciated that even more rapid acting (“ultra rapid acting”) insulins can be achieved by removing the zinc cations from insulin 15 altogether. Unfortunately, the consequence of the hexamer dissociation is typically a considerable impairment in insulin stability both with respect to physical stability (e.g. stability to aggregation) and chemical stability (e.g. stability to deamidation). For example, monomeric insulin or insulin analogues having a rapid onset of action are known to aggregate and become physically unstable very rapidly because the 20 formation of insoluble aggregates proceeds via monomers of insulin. Various approaches to addressing this problem have been described in the art:
US5,866,538 (Norup) describes insulin preparations of superior chemical stability comprising human insulin or an analogue or derivative thereof, glycerol 25 and/or mannitol and 5 mM to 100 mM of a halogenide (e.g. NaCl).
US7,205,276 (Boderke) addresses the stability problems associated with preparing zinc-free formulations of insulin and insulin derivatives and analogues and describes an aqueous liquid formulation comprising at least one insulin derivative, at 30 least one surfactant, optionally at least one preservative and optionally at least one of an isotonicizing agent, a buffer and an excipient, wherein the formulation is stable and free from or contains less than 0.4% (e.g. less than 0.2%) by weight of zinc based on the insulin content of the formulation. The preferred surfactant appears to be polysorbate 20 (polyoxyethylene (20) sorbitan monolaurate). 35
US2008/0194461 (Maggio) describes formulations of peptides and polypeptides including insulin which contain an alkyl glycoside, which component is said to reduce aggregation and immunogenicity.
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WO2012/006283 (Pohl) describes formulations containing insulin together with a zinc chelator such as ethylenediaminetetraacetate (EDTA). Modulating the type and quantity of EDTA is said to change the insulin absorption profile. Calcium EDTA is the preferred form of EDTA since it is said to be associated with reduced pain at the injection site and is less likely to remove calcium from the body. 5 Preferred formulations also contain citrate which is said to further enhance absorption and to improve the chemical stability of the formulation.
US2010/0227795 (Steiner) describes a composition comprising insulin, a dissociating agent such as citric acid or sodium citrate, and a zinc chelator such as 10 EDTA wherein the formulation has a physiological pH and is a clear aqueous solution. The formulations are said to have improved stability and rapid onset of action.
WO2015/120457 (Wilson) describes stabilized ultra-rapid acting insulin 15 formulations comprising insulin in combination with a zinc chelator such as EDTA, a dissolution/stabilization agent such as citric acid, a magnesium salt, a zinc compound and optionally additional excipients.
Further approaches to accelerating the absorption and effect of insulin 20 through the use of specific accelerating additives have been described:
WO91/09617 (Jørgensen) reports that nicotinamide or nicotinic acid or a salt thereof increases the speed of absorption of insulin from aqueous preparations administered parenterally. 25
WO2010/149772 (Olsen) describes a formulation comprising insulin, a nicotinic compound and arginine. The presence of arginine is said to improve the chemical stability of the formulation.
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WO2015/171484 (Christe) describes rapid-acting formulations of insulin wherein onset of action and/or absorption of insulin is faster due to the presence of treprostinil.
US2013/0231281 (Soula) describes an aqueous solution composition 35 comprising insulin or an insulin analogue and at least one oligosaccharide whose average degree of polymerisation is between 3 and 13 and whose polydispersity index is above 1.0, said oligosaccharide having partially substituted carboxyl functional groups, the unsubstituted carboxyl functional groups being salifiable. Such a formulation is said to be rapid acting. 40
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WO2017/191464 (Arecor Limited) describes an aqueous liquid pharmaceutical formulation comprising insulin or an insulin analogue, ionic zinc, a chelating agent and polysorbate 80.
WO2016/100042 (Eli Lilly and Company) describes a composition of human 5 insulin or insulin analogue that includes specific concentrations of citrate, chloride, in some cases including the addition of sodium chloride, zinc and, optionally magnesium chloride and/or surfactant, said to have faster pharmacokinetic and/or pharmacodynamic action than commercial formulations of existing insulin analogue products. 10
Commercially available rapid-acting insulin formulations are available as 100 U/ml formulations (HUMALOG® (insulin lispro), NOVORAPID® (also known as NOVOLOG®, insulin aspart) and APIDRA® (insulin glulisine)) and 200 U/ml formulations (HUMALOG®). 15
There are a number of devices that can be used to deliver insulin, including syringes, insulin pumps and insulin pens. Syringes can typically be used to deliver basal (long-acting) insulins, typically 20 as one injection per day. Whilst syringes are still used, they are gradually being replaced by more convenient insulin pens. Insulin pens are a very convenient way of delivering both basal and prandial insulin. Insulin pens contain a cartridge that is filled with insulin and an apparatus for 25 dispensing a required amount of insulin, as needed by the user. The required amount is first selected (this often referred to as being “dialed”) using a specifically designed mechanism and then dispensed via a very small retractable needle whilst holding the pen against the body (typically the abdomen).
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It would be desirable if an injection pen system were available which can deliver compositions of insulin or insulin analogues from a reservoir, which are rapid or ultra-rapid acting, and which remain stable upon storage and in-use. It would be particularly desirable, particularly for diabetic patients that require large doses of insulin, if an injection pen system were available which can deliver high strength 35 compositions of insulin or insulin analogues that are rapid or ultra-rapid acting, and which remain stable upon storage and in-use.
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SUMMARY OF THE INVENTION
According to the invention there is provided an injection pen system comprising an injector mechanism and a reservoir comprising an aqueous liquid pharmaceutical composition for delivery by means of said injector mechanism to a mammal wherein the composition comprises (i) an insulin compound, (ii) ionic zinc 5 and (iii) an alkyl glycoside as a non-ionic surfactant. The compositions of the system of the invention provide insulin in a form with good physical and chemical stability, preferably in a form which is rapid or ultra-rapid acting. The present inventors have importantly identified that use of an alkyl glycoside as a non-ionic surfactant increases the storage stability of insulin compositions, which is expected to permit 10 the use of an injection pen based system to deliver aqueous liquid pharmaceutical compositions of insulin to the body of a mammal from one or more reservoirs with good in-use stability.
As noted in the background discussion above, use of EDTA to chelate zinc 15 ions in hexameric insulin does increase the rapidity of action but at the cost of greatly reduced stability. Without being limited by theory, the present inventors have also appreciated that the use in certain embodiments of the invention of zinc together with species which bind zinc less strongly can achieve similar effects in terms of speed of action and their moderately destabilising effects can be reduced or eliminated by 20 using a non-ionic surfactant. The present inventors have further appreciated that the presence of such a zinc binding species accelerates the onset of action of a high concentration (high strength) insulin compound composition thereby mitigating the delaying effect on insulin onset of action which has been observed when the concentration of insulin compound in a composition is increased. 25
Compositions of the system of the invention may be used in the treatment of subjects suffering from diabetes mellitus, particularly Type 1 diabetes mellitus especially for administration at meal times.
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As can be seen from the accompanying examples, example compositions of the system of the invention are significantly more stable than compositions without alkyl glycoside as non-ionic surfactant including under stress conditions that model those of an injection pen system. The example compositions achieve a rapid speed of action of insulin and are more stable than prior art rapid acting insulin formulations 35 containing EDTA. Furthermore, example compositions of the system of the invention contain high concentrations of insulin compound while maintaining a rapid onset of action.
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DESCRIPTION OF THE SEQUENCE LISTING
SEQ ID NO: 1: A chain of human insulin
SEQ ID NO: 2: B chain of human insulin
SEQ ID NO: 3: B chain of insulin lispro
SEQ ID NO: 4: B chain of insulin aspart 5
SEQ ID NO: 5: B chain of insulin glulisine
FIGURES
Fig. 1. Pharmacodynamic profiles of Formulations 4A-4C of Example 4 in a validated diabetic Yucatan miniature pig model. 10
Fig. 2. Pharmacodynamic profile of Formulations 13A and 13B of Example 13 in a validated diabetic Yucatan miniature pig model.
Fig. 3. Pharmacodynamic profiles of formulations 14A-14D of Example 14 in a 15 validated diabetic Yucatan miniature pig model.
Fig. 4. Pharmacokinetic profiles of formulations 14A-14C of Example 14 in a validated diabetic Yucatan miniature pig model.
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Fig. 5. Pharmacodynamic profiles of formulations 15A-15D of Example 15 in a validated diabetic Yucatan miniature pig model.
Fig. 6. Pharmacokinetic profiles of formulations 15A, 15B and 15D of Example 15 in a validated diabetic Yucatan miniature pig model. 25
DETAILED DESCRIPTION OF THE INVENTION
As used herein, “insulin compound” refers to insulin and insulin analogues.
As used herein, “insulin” refers to native human insulin having an A chain and a B chain as set out in SEQ ID NOS: 1 and 2 and containing and connected by 30 disulfide bridges as in the native molecule (Cys A6-Cys A11, Cys B7 to Cys A7 and Cys-B19-Cys A20). Insulin is suitably recombinant insulin.
“Insulin analogue” refers to an analogue of insulin which is an insulin receptor agonist and has a modified amino acid sequence, such as containing 1 or 2 amino 35 acid changes in the sequence of the A or B chain (especially the B chain). Desirably such amino acid modifications are intended to reduce affinity of the molecule for zinc and thus increase speed of action. Thus, desirably an insulin analogue has a speed of action which is the same as or preferably greater than that of insulin. The speed of action of insulin or an insulin analogue may be determined in the Diabetic Pig 40
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Pharmacokinetic/Pharmacodynamic Model (see Examples, General Methods (c)). Exemplary insulin analogues include faster acting analogues such as insulin lispro, insulin aspart and insulin glulisine. These forms of insulin have the human insulin A chain but variant B chains – see SEQ ID NOS: 3-5. Further faster acting analogues are described in EP0214826, EP0375437 and EP0678522 the contents of which are 5 herein incorporated by reference in their entirety. Suitably, the insulin compound is not insulin glargine. Suitably, the insulin compound is not insulin degludec. Suitably, the insulin compound is a rapid-acting insulin compound, wherein “rapid-acting” is defined as an insulin compound which has a speed of action which is greater than that of native human insulin, e.g. as measured using the Diabetic Pig 10 Pharmacokinetic/Pharmacodynamic Model (see Examples, General Methods (c)).
In one embodiment, the insulin compound is recombinant human insulin. In another embodiment, it is insulin lispro. In another embodiment, it is insulin aspart. In another embodiment, it is insulin glulisine. In another embodiment, the insulin 15 compound is not recombinant human insulin.
The term “aqueous liquid pharmaceutical composition”, as used herein, refers to a composition suitable for therapeutic use in which the aqueous component is or comprises water, preferably distilled water, deionized water, water for injection, 20 sterile water for injection or bacteriostatic water for injection. The aqueous liquid pharmaceutical compositions of the system of the invention are solution compositions in which all components are dissolved in water.
The concentration of insulin compound in the composition is in the range 10-25 1000 U/ml e.g. 50-1000 U/ml, e.g. 400-1000 U/ml, e.g. 500-1000 U/ml, e.g. 600-1000 U/ml, e.g. 700-1000 U/ml, e.g. 800-1000 U/ml, e.g. 900-1000 U/ml, e.g. 1000U/ml. In one embodiment, the concentration of insulin compound in the composition is 10-250 U/ml.
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“U/ml” as used herein describes the concentration of insulin compound in terms of a unit per volume, wherein “U” is the international unit of insulin activity (see e.g. European Pharmacopoeia 5.0, Human Insulin, pp 1800-1802).
The compositions of the system of the invention contain ionic zinc i.e. Zn2+ 35 ions. The source of the ionic zinc will typically be a water-soluble zinc salt such as ZnCl2, ZnO, ZnSO4, Zn(NO3)2 or Zn(acetate)2 and most suitably ZnCl2 or ZnO.
The ionic zinc in the composition is typically present at a concentration of more than 0.05% e.g. more than 0.1% e.g. more than 0.2%, more than 0.3% or more 40
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than 0.4% by weight of zinc based on the weight of insulin compound in the composition. Thus, the concentration of the ionic zinc in the composition may be more than 0.5% by weight of zinc based on the weight of insulin compound in the composition, for example 0.5-1%, e.g. 0.5-0.75%, e.g. 0.5-0.6% by weight of zinc based on the weight of insulin compound in the composition. For the purpose of the 5 calculation the weight of the counter ion to zinc is excluded.
In a composition e.g. containing 1000 U/ml of insulin compound the concentration of the ionic zinc will typically be more than 0.15 mM e.g. more than 0.3 Mm, e.g. more than 0.6 mM, more than 0.9 mM or more than 1.2 mM. Thus, the 10 concentration of the ionic zinc in the composition may be more than 1.5 mM, for example 1.5-6.0 mM, e.g. 2.0-4.5 mM, e.g. 2.5-3.5 mM.
The compositions of the system of the invention may optionally comprise a zinc binding species e.g. at a concentration of 1 mM or more and, for example, 15 selected from species having a logK with respect to zinc ion binding in the range 4.5-12.3 at 25 °C. Suitably, the zinc binding species is selected from species having a logK with respect to zinc ion binding in the range 4.5-10 at 25 °C. Metal binding stability constants listed in the National Institute of Standards and Technology reference database 46 (Critically Selected Stability Constants of Metal Complexes) 20 can be used. The database typically lists logK constants determined at 25 °C. Therefore, the suitability of a zinc binding species for the present invention can be determined based on its logK metal binding stability constant with respect to zinc binding, as measured at 25 °C and as quoted by the database. The zinc binding species may also be described as an “accelerator” in the compositions according to 25 the invention. Exemplary zinc binding species include polydendate organic anions. Thus, in a preferred embodiment, the zinc binding species is citrate (logK = 4.93) which can, for example, be employed as trisodium citrate or citric acid. Further examples include pyrophosphate (logK = 8.71), aspartate (logK = 5.87), glutamate (logK = 4.62), cysteine (logK = 9.11), cystine (logK = 6.67) and glutathione (logK = 30 7.98). Other possible zinc binding species include substances that can contribute a lone pair of electrons or electron density for interaction with ionic zinc such as polydendate amines including ethylenediamine (logK = 5.69), diethylenetriamine (DETA, logK = 8.88) and triethylenetetramine (TETA, logK = 11.95); and aromatic or heteroaromatic substances that can contribute a lone pair of electrons especially 35 those comprising an imidazole moiety such as histidine (logK = 6.51). Thus, in one embodiment, the zinc binding species having a logK with respect to zinc ion binding in the range 4.5-12.3 is selected from citrate, pyrophosphate, aspartate, glutamate, cysteine, cystine, glutathione, ethylenediamine, histidine, DETA and TETA.
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The most suitable concentration of the zinc binding species will depend on the agent and its logK value and will typically be in the range 1-100 mM. The concentration of zinc binding species can be adjusted according to the particular concentration of insulin compound present in the composition, in order to provide the desired accelerating effect. 5
For example, the zinc binding species having a logK with respect to zinc ion binding in the range 4.5-12.3 may be present at a concentration of 1-60 mM. Suitably the concentration of the zinc binding species in the composition is 5-60 mM e.g. 5-60 mM, e.g. 10-60 mM, e.g. 20-60 mM, e.g. 30-60 mM, e.g. 40-60 mM, e.g. 40-50 mM, 10 more preferably around 44 mM when the zinc binding species is citrate or histidine for insulin compound 1000 U/ml compositions.
Anionic zinc binding species may be employed as the free acid or a salt form, such as a salt form with sodium or calcium ions, especially sodium ions. 15
A mixture of zinc binding species may be employed, although a single zinc binding species is preferred.
Suitably the molar ratio of ionic zinc to zinc binding species in the 20 composition is 1:3 to 1:175.
The following ranges are particularly of interest especially for citrate or histidine as zinc binding species: e.g. 1:10-1:175, e.g. 1:10 to 1:100, e.g. 1:10-1:50, e.g. 1:10 to 1:30, e.g. 1:10 to 1:20 (especially for insulin compound 1000 U/ml 25 composition).
For example, a composition containing 1000 U/ml of insulin compound may contain around 3 mM of ionic zinc (i.e. around 197 μg/ml of ionic zinc, i.e. around 0.54% by weight of zinc based on the weight of insulin compound in the composition) 30 and around 30-60 mM e.g. 40-60 mM e.g. 40-50 mM zinc binding species (especially citrate).
In one embodiment, the ratio of insulin compound concentration (U/ml) to zinc binding species (mM) in the composition is in the range 100:1 to 2:1 e.g. 50:1 to 35 2:1, e.g. 40:1 to 2:1.
In one embodiment, the composition is substantially free of EDTA and any other zinc binding species having a logK with respect to zinc binding of more than 12.3 as determined at 25 °C. In an embodiment, the formulations of the invention 40
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are substantially free of EDTA (logK = 14.5). Further examples of zinc binding species which have a logK metal binding stability constant with respect to zinc binding of more than 12.3 to be avoided include EGTA (logK = 12.6). In general, the composition of the system of the invention will be substantially free of tetradentate ligands or ligands of higher denticity. In an embodiment, the composition of the 5 system of the invention is substantially free of zinc binding species having a logK with respect to zinc ion binding of 10-12.3 at 25 °C. “Substantially free” means that the concentration of zinc binding species which have a logK metal binding stability constant with respect to zinc binding as specified (such as EDTA) is less than 0.1 mM, such as less than 0.05 mM, such as less than 0.04 mM or less than 0.01 mM. 10
Where present, zinc ion binding species which have acid forms (e.g. citric acid) may be introduced into the aqueous compositions of the system of the invention in the form of a salt of the acid, such as a sodium salt (e.g. trisodium citrate). Alternatively, they can be introduced in the form of the acid with subsequent 15 adjustment of pH to the required level. The present inventors have found that in some circumstances introducing the acid form (such as citric acid) into the composition instead of the salt form (e.g. trisodium citrate) may have advantages in terms of providing superior chemical and physical stability. Thus, in an embodiment, the source of the citrate as zinc ion binding species is citric acid. 20
In an embodiment, the composition comprises (i) an insulin compound (e.g. an insulin compound other than insulin glargine), (ii) ionic zinc, (iii) a zinc binding species selected from diethylenetriamine (DETA) and triethylenetetramine (TETA), and (iv) an alkyl glycoside as non-ionic surfactant. Such a composition may, for 25 example. be substantially free of ethylenediaminetetraacetate (EDTA) and any other zinc binding species having a logK with respect to zinc ion binding of more than 12.3 at 25 °C. The zinc binding species may, for example, be present at a concentration of about 0.05 mM or more e.g. 0.05-5 mM e.g. 0.05-2 mM. The molar ratio of ionic zinc to the zinc binding species in the composition may, for example, be 2:1 to 1:10. 30
In an embodiment, the composition comprises (i) an insulin compound,
(ii) ionic zinc, (iii) a zinc binding species at a concentration of 1 mM or more selected from species having a logK with respect to zinc ion binding in the range 4.5-10 at 25 °C, (iv) a zinc binding species selected from species having a logK with respect to 35 zinc ion binding of more than 12.3 at 25 °C at a concentration of less than about 0.3 mM, and (v) an alkyl glycoside as non-ionic surfactant. In an embodiment, the zinc binding species having a logK with respect to zinc ion binding of more than 12.3 at 25 °C is present in the composition at a concentration of between about 0.01 mM and about 0.3 mM. In an embodiment, the zinc binding species having a logK with 40
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respect to zinc ion binding of more than 12.3 at 25 °C is selected from ethylenediaminetetraacetate (EDTA), ethyleneglycoltetraacetate (EGTA), tetraethylenepentamine, N-(2-hydroxyethyl)ethylenedinitrilotriacetate (HEDTA), 1-methyl-ethylenedinitrilotriacetate (PDTA), 1-ethyl-ethylenedinitrilotriacetate, 1-propyl- thylenedinitrilotriacetate, 1-carboxyethylene-ethylenedinitrilotriacetate, 5 triethylenetetranitrilohexaacetate, tetraethylenepentanitriloheptaacetate (TPHA) and tris(2-aminoethyl)amine (Tren), and especially is EDTA. For example, the molar ratio of ionic zinc to EDTA as zinc binding species having a logK with respect to zinc ion binding of more than 12.3 at 25 °C is 2:1 to 25:1. In an embodiment, the zinc binding species having a logK with respect to zinc ion binding in the range 4.5-10 at 10 25 °C is selected from citrate, pyrophosphate, aspartate, glutamate, cysteine, cystine, glutathione, ethylenediamine and histidine and especially is citrate. In an embodiment, the zinc binding species having a logK with respect to zinc ion binding in the range 4.5-10 at 25 °C is present at a concentration of 1-50 mM. In an embodiment, the molar ratio of ionic zinc to zinc binding species having a logK with 15 respect to zinc ion binding in the range 4.5-10 at 25 °C is 1:3 to 1:500.
The compositions of the system of the invention contain an alkyl glycoside as a non-ionic surfactant. In one embodiment, the alkyl glycoside is selected from the group consisting of dodecyl maltoside, dodecyl glucoside, octyl glucoside, octyl 20 maltoside, decyl glucoside, decyl maltoside, decyl glucopyranoside, tridecyl glucoside, tridecyl maltoside, tetradecyl glucoside, tetradecyl maltoside, hexadecyl glucoside, hexadecyl maltoside, sucrose monooctanoate, sucrose monodecanoate, sucrose monododecanoate, sucrose monotridecanoate, sucrose monotetradecanoate and sucrose monohexadecanoate. In one embodiment, the 25 alkyl glycoside is dodecyl maltoside or decyl glucopyranoside. In one preferred embodiment, the alkyl glycoside is dodecyl maltoside.
The concentration of the alkyl glycoside in the composition will typically be in the range 1-1000 μg/ml, e.g. 5-500 μg/ml, e.g. 10-200 μg/ml, such as 10-100 μg/ml 30 or around 50 μg/ml. In one embodiment, the non-ionic surfactant is present at a concentration of 10-400 μg/ml e.g. 20-400 μg/ml, 50-400 μg/ml, 10-300 μg/ml, 20-300 μg/ml, 50-300 μg/ml, 10-200 μg/ml, 20-200 μg/ml, 50-200 μg/ml, 10-100 μg/ml, 20-100 μg/ml or 50-100 μg/ml.
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In another embodiment, the concentration of insulin compound is 800-1000 U/ml and the non-ionic surfactant is present at a concentration of 50-200 μg/ml. In this embodiment, suitably the non-ionic surfactant is dodecyl maltoside.
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In one embodiment, the composition of the system of the invention comprises (i) an insulin compound at a concentration of 50-500 U/ml (ii) ionic zinc, (iii) optionally citrate as a zinc binding species at a concentration of 1 mM or more, and (iv) a non-ionic surfactant which is an alkylglycoside; and wherein the composition is substantially free of EDTA and any other zinc binding species having a logK with 5 respect to zinc ion binding of more than 12.3 at 25 ° C. Suitably, the citrate may be present in the composition at a concentration of 10-30 mM e.g. 10-20 mM e.g. 15-25 mM e.g. 20-30 mM.
Suitably the pH of the composition of the system of the invention is in the 10 range 5.5-9.0 e.g. in the range 7.0-7.5. In order to minimise injection pain, the pH is preferably close to physiological pH (around pH 7.4). In one embodiment of the system of the invention, the pH is in the range 7.0-8.0 e.g. 7.5. In another embodiment of the system, the pH is in the range 7.6-8.0 e.g. 7.8.
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Suitably, the composition of the system of the invention comprises a buffer (e.g. one or more buffers) in order to stabilise the pH of the composition, which can also be selected to enhance protein stability. In one embodiment, a buffer is selected to have a pKa close to the pH of the composition; for example, histidine is suitably employed as a buffer when the pH of the composition is in the range 5.0-7.0. 20 Such a buffer may be employed in a concentration of 0.5-20 mM e.g. 2-5 mM. If histidine is included in the composition as a zinc binding species it will also have a buffering role at this pH. In another embodiment, the composition comprises a phosphate buffer. Sodium phosphate is suitably employed as a buffer when the pH of the composition is in the range 6.1-8.1. Such a buffer may be employed in a 25 concentration of 0.5-20 mM e.g. 2-5 mM, e.g. 2 mM. Alternatively, in another embodiment, the composition of the system of the invention is further stabilised as disclosed in WO2008/084237 (herein incorporated by reference in its entirety), which describes a composition comprising a protein and one or more additives, characterised in that the system is substantially free of a conventional buffer, i.e. a 30 compound with an ionisable group having a pKa within 1 unit of the pH of the composition at the intended temperature range of storage of the composition, such as 25 °C. In this embodiment, the pH of the composition is set to a value at which the composition has maximum measurable stability with respect to pH; the one or more additives (displaced buffers) are capable of exchanging protons with the insulin 35 compound and have pKa values at least 1 unit more or less than the pH of the composition at the intended temperature range of storage of the composition. The additives may have ionisable groups having pKa between 1 to 5 pH units, preferably between 1 to 3 pH units, most preferably from 1.5 to 2.5 pH units, of the pH of the aqueous composition at the intended temperature range of storage of the 40
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composition (e.g. 25 °C). Such additives may typically be employed at a concentration of 0.5-10 mM e.g. 2-5 mM.
The compositions of the system cover a wide range of osmolarity, including hypotonic, isotonic and hypertonic compositions. Preferably, the composition of the 5 system of the invention is substantially isotonic. Suitably the osmolarity of the composition is selected to minimize pain according to the route of administration e.g. upon injection. Preferred compositions have an osmolarity in the range of about 200 to about 500 mOsm/L. Preferably, the osmolarity is in the range of about 250 to about 350 mOsm/L. More preferably, the osmolarity is about 300 mOsm/L. 10
Tonicity of the composition may be adjusted with a tonicity modifying agent (e.g. one or more tonicity modifying agents). Thus, the composition of the system of the invention may further comprise a tonicity modifying agent (e.g. one or more tonicity modifying agents). The tonicity modifying agent may be charged or 15 uncharged. Examples of charged tonicity modifying agents include salts such as a combination of sodium, potassium, magnesium or calcium ions, with chloride, sulfate, carbonate, sulfite, nitrate, lactate, succinate, acetate or maleate ions (especially sodium chloride or sodium sulphate, particularly sodium chloride).
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In one embodiment, the charged tonicity modifying agent is sodium chloride. The insulin compound compositions of the system of the invention may contain a residual NaCl concentration of 2-4 mM as a result of the use of standard acidification and subsequent neutralization steps employed in preparing insulin compositions. Amino acids such as arginine, glycine or histidine may also be used for this purpose. 25 Charged tonicity modifying agent (e.g. NaCl) may be used at a concentration of 100–300 mM, e.g. around 150 mM. Preferably, the chloride is present at a concentration of >60 mM e.g. >65 mM, >75 mM, >80 mM, >90 mM, >100 mM, >120 mM or >140 mM.
30
Suitably an uncharged rather than a charged tonicity modifying agent is used when the concentration of insulin compound in the composition is 400 U/ml or more.
Examples of uncharged tonicity modifying agents include sugars, sugar alcohols and other polyols, such as trehalose, sucrose, mannitol, glycerol, 1,2-35 propanediol, raffinose, lactose, dextrose, sorbitol or lactitol (especially trehalose, mannitol, glycerol or 1,2-propanediol, particularly glycerol). In one embodiment, the uncharged tonicity modifying agent is selected from the group consisting of trehalose, mannitol, glycerol and 1,2-propanediol. In another embodiment, the uncharged tonicity modifying agent is glycerol. Uncharged tonicity modifying agent is 40
15
preferably used at a concentration of 200–500 mM, e.g. around 300 mM. Another range of interest is 100-500 mM. In one embodiment, the uncharged tonicity modifying agent in the composition is at a concentration of 100–300 mM, e.g. 150–200 mM, 170-180 mM or around 174 mM. In one embodiment, the uncharged tonicity modifying agent in the composition is glycerol at a concentration of 100–300 5 mM, e.g. 150-200 mM, 170-180 mM or around 174 mM.
In one embodiment, the composition of the system of the invention comprises <10 mM chloride (e.g. sodium chloride), for example <9 mM, <8 mM, <7 mM, <6 mM or <5 mM, or is substantially free of chloride (e.g. sodium chloride) i.e. no chloride is 10 added to the composition beyond any chloride that may be contributed as part of pH adjustment.
When the insulin compound is insulin lispro, the tonicity is suitably adjusted using an uncharged tonicity modifying agent, preferably at a concentration of 200–15 500 mM, e.g. around 300 mM. In this embodiment, the uncharged tonicity modifying agent is suitably selected from the group consisting of trehalose, mannitol, glycerol and 1,2-propanediol (most suitably glycerol). In another embodiment, the uncharged tonicity modifying agent is used at a concentration of 100–300 mM, e.g. 150–200 mM, 170-180 mM or around 174 mM. In one embodiment, the uncharged tonicity 20 modifying agent is glycerol at a concentration of 100–300 mM, e.g. 150-200 mM, 170-180 mM or around 174 mM.
When the insulin compound is insulin aspart, the tonicity is suitably adjusted using an uncharged tonicity modifying agent, preferably at a concentration of 200–25 500 mM, e.g. around 300 mM. In this embodiment, the uncharged tonicity modifying agent is suitably selected from the group consisting of trehalose, mannitol, glycerol and 1,2-propanediol (most suitably glycerol). In another embodiment, the uncharged tonicity modifying agent is used at a concentration of 100–300 mM, e.g. 150–200 mM, 170-180 mM or around 174 mM. In one embodiment, the uncharged tonicity 30 modifying agent is glycerol at a concentration of 100–300 mM, e.g. 150-200 mM, 170-180 mM or around 174 mM.
When the insulin compound is insulin glulisine, the tonicity is suitably adjusted using an uncharged tonicity modifying agent, preferably at a concentration 35 of 200–500 mM, e.g. around 300 mM. In this embodiment, the uncharged tonicity modifying agent is suitably selected from the group consisting of trehalose, mannitol, glycerol and 1,2-propanediol (most suitably glycerol). In another embodiment, the uncharged tonicity modifying agent is used at a concentration of 100–300 mM, e.g. 150–200 mM, 170-180 mM or around 174 mM. In one embodiment, the uncharged 40
16
tonicity modifying agent is glycerol at a concentration of 100–300 mM, e.g. 150-
200 mM, 170-180 mM or around 174 mM.
The ionic strength of a composition of the system of the invention may be
calculated according to the formula I:
=
= 
n
X 1
2
x x I 0.5 c z
5
in which cx is molar concentration of ion x (mol L-1), zx is the absolute value of the
charge of ion x and the sum covers all ions (n) present in the composition, wherein
the contribution of the insulin compound and zinc binding species (if present) should
be ignored for the purposes of the calculation. The contribution of ionic zinc should
10 be included for the purposes of the calculation. For zwitterions, the absolute value of
the charge is the total charge excluding polarity, e.g. for glycine the possible ions
have absolute charge of 0, 1 or 2 and for aspartate the possible ions have absolute
charge of 0, 1, 2 or 3.
15 In an embodiment, and particularly when the concentration of insulin
compound in the composition is 400 U/ml or more, the ionic strength of the
composition is suitably less than 40 mM, less than 30 mM, less than 20 mM or less
than 10 mM.
20 In one embodiment the composition of the system of the invention comprises
(i) an insulin compound at a concentration of 400-1000 U/ml e.g. 500-1000 U/ml (ii)
ionic zinc, (iii) optionally citrate as a zinc binding species at a concentration of 1 mM
or more, and (iv) an alkyl glycoside as a non-ionic surfactant; wherein the
composition is substantially free of EDTA and any other zinc binding species having
25 a logK with respect to zinc ion binding of more than 12.3 at 25 °C, and wherein the
ionic strength of the composition is less than 40 mM, said ionic strength being
calculated using the formula I:
=
= 
n
X 1
2
x x I 0.5 c z
in which cx is molar concentration of ion x (mol L-1), zx is the absolute value of the
30 charge of ion x and the sum covers all ions (n) present in the composition, wherein
the contribution of the insulin compound and zinc binding species (if present) should
be ignored for the purposes of the calculation. The contribution of ionic zinc should
be included for the purposes of the calculation. Suitably, the citrate is present in the
composition at a concentration of 30-60 mM e.g. 30-50 mM e.g. 40-50 mM.
35
17
In another embodiment, the composition of the system of the invention comprises (i) an insulin compound at a concentration of 400-1000 U/ml e.g. 500-1000 U/ml (ii) ionic zinc, (iii) optionally citrate as a zinc binding species at a concentration of 1 mM or more, and (iv) a non-ionic surfactant which is an alkyl glycoside; and wherein the composition is substantially free of EDTA and any other 5 zinc binding species having a logK with respect to zinc ion binding of more than 12.3 at 25 °C. Suitably, the citrate may be present in the composition at a concentration of 30-60 mM e.g. 30-50 mM, e.g. 30-40 mM, e.g. 35-45 mM, e.g. 40-50 mM. Suitably the ionic strength of the composition is less than 40 mM calculated using Formula I. Suitably, the formulation of the invention comprises <10 mM chloride 10 (e.g. sodium chloride), for example <9 mM, <8 mM, <7 mM, <6 mM or <5 mM, or is substantially free of chloride (e.g. sodium chloride) i.e. no chloride is added to the formulation beyond any chloride that may be contributed as part of pH adjustment. In one embodiment, the composition comprises an uncharged tonicity modifying agent. 15
In one embodiment, the insulin compound is present at a concentration of 400-1000 U/ml e.g. >400-1000 U/ml, 500-1000 U/ml, >500-1000 U/ml, 600-1000 U/ml, >600-1000 U/ml, 700-1000 U/ml, >700-1000 U/ml, 750-1000 U/ml, >750-1000 U/ml, 800-1000 U/ml, >800-1000 U/ml, 900-1000 U/ml, >900-1000 U/ml or 1000 20 U/ml, and the ionic strength taking account of ions in the composition except for the zinc binding species, the insulin compound and the ionic zinc is less than 30 mM, e.g. less than 20 mM, e.g. less than 10 mM such as 1-10 mM. In a further embodiment, the ionic strength taking account of ions in the composition except for the zinc binding species, the insulin compound and the ionic zinc is less than 25 mM, 25 less than 20 mM, less than 15 mM, or less than 10 mM, or is in the range 5-<30 mM, 5-30 mM, 5-20 mM, 2-20 mM, 1-10 mM, 2-10 mM or 5-10 mM.
When the insulin compound is insulin lispro a concentration of 400-1000 U/ml e.g. >400-1000 U/ml, 500-1000 U/ml, >500-1000 U/ml, 600-1000 U/ml, >600-1000 30 U/ml, 700-1000 U/ml, >700-1000 U/ml, 750-1000 U/ml, >750-1000 U/ml, 800-1000 U/ml, >800-1000 U/ml, 900-1000 U/ml, >900-1000 U/ml or 1000 U/ml, the ionic strength of the composition is suitably kept to a minimum level since higher ionic strength compositions are less stable than lower ionic strength compositions, particularly at high concentrations of insulin. Suitably the ionic strength taking 35 account of ions in the composition except for the zinc binding species, the insulin compound and the ionic zinc is less than 30 mM, e.g. less than 20 mM, e.g. less than 10 mM such as 1-10 mM. In particular, the ionic strength taking account of ions in the composition except for the zinc binding species, the insulin compound and the ionic zinc is less than 25 mM, less than 20 mM, less than 15 mM, or less than 10 40
18
mM, or is in the range 5-<30 mM, 5-30 mM, 5-20 mM, 2-20 mM, 1-10 mM, 2-10 mM or 5-10 mM.
When the insulin compound is insulin aspart at a concentration of 400-1000 U/ml e.g. >400-1000 U/ml, 500-1000 U/ml >500-1000 U/ml, 600-1000 U/ml, >600-5 1000 U/ml, 700-1000 U/ml, >700-1000 U/ml, 750-1000 U/ml, >750-1000 U/ml, 800-1000 U/ml, >800-1000 U/ml, 900-1000 U/ml, >900-1000 U/ml or 1000 U/ml, the ionic strength of the composition is suitably kept to a minimum level since higher ionic strength compositions are less stable than lower ionic strength compositions. Suitably the ionic strength taking account of ions in the composition except for the 10 zinc binding species, the insulin compound and the ionic zinc is less than 30 mM, e.g. less than 20 mM, e.g. less than 10 mM. In particular, the ionic strength taking account of ions in the composition except for the zinc binding species, the insulin compound and the ionic zinc is less than 25 mM, less than 20 mM, less than 15 mM, or less than 10 mM, or is in the range 5-<30 mM, 5-30 mM, 5-20 mM, 2-20 mM, 1-10 15 mM, 2-10 mM or 5-10 mM. The tonicity may suitably be adjusted using an uncharged tonicity modifying agent.
When the insulin compound is insulin glulisine at a concentration of 400-1000 U/ml e.g. >400-1000 U/ml, 500-1000 U/ml >500-1000 U/ml, 600-1000 U/ml, >600-20 1000 U/ml, 700-1000 U/ml, >700-1000 U/ml, 750-1000 U/ml, >750-1000 U/ml, 800-1000 U/ml, >800-1000 U/ml, 900-1000 U/ml, >900-1000 U/ml or 1000 U/ml, the ionic strength of the composition is suitably kept to a minimum level since higher ionic strength compositions may be less stable than lower ionic strength compositions. Suitably the ionic strength taking account of ions in the composition except for the 25 zinc binding species, the insulin compound and the ionic zinc is less than 30 mM, e.g. less than 20 mM, e.g. less than 10 mM. In particular, the ionic strength taking account of ions in the composition except for the zinc binding species, the insulin compound and ionic zinc is less than 25 mM, less than 20 mM, less than 15 mM, or less than 10 mM, or is in the range 5-<30 mM, 5-30 mM, 5-20 mM, 2-20 mM, 1-10 30 mM, 2-10 mM or 5-10 mM.
The composition of the system of the invention may optionally further comprise a preservative (e.g. one or more preservatives). One or more preservatives may be employed. In one embodiment, the preservative is selected 35 from the group consisting of phenol, m-cresol, chlorocresol, benzyl alcohol, propylparaben, methylparaben, benzalkonium chloride and benzethonium chloride.
The composition of the system of the invention may optionally further comprise nicotinamide. The presence of nicotinamide may further increase the 40
19
speed of onset of action of insulin formulated in compositions of the system of the invention. Suitably, the concentration of nicotinamide is in the range 10-150 mM, preferably in the range 20-100 mM, such as around 80 mM.
The composition of the system of the invention may optionally further 5 comprise nicotinic acid or a salt thereof. The presence of nicotinic acid or a salt thereof may also further increase the speed of onset of action of insulin formulated in compositions of the system of the invention. Suitably, the concentration of nicotinic acid or a salt thereof is in the range 10-150 mM, preferably in the range 20-100 mM, such as around 80 mM. Example salts include metal salts such as sodium, 10 potassium and magnesium salts.
Typically, one of nicotinamide and nicotinic acid (or as salt thereof) may be included in the composition but not both.
15
In an embodiment, the composition comprises (i) an insulin compound, (ii) ionic zinc, (iii) a nicotinic compound, (iv) an alkyl glycoside as a non-ionic surfactant; and (v) a salt selected from the salts formed between Group 1 metals and a mono or divalent anion. In an embodiment, the nicotinic compound is nicotinamide or nicotinic acid or a salt thereof. In an embodiment, the nicotinic compound is present in the 20 composition at a concentration of 10-150 mM. In an embodiment, the Group 1 metal is sodium. In an embodiment, the salt is the sodium salt of a mono or divalent anion. In an embodiment, the anion is chloride or acetate. Thus, for example, the salt is sodium chloride or sodium acetate. In an embodiment, the salt is present in the composition at a concentration of 30-200 mM. 25
The composition of the system of the invention may optionally further comprise treprostinil or a salt thereof. The presence of the treprostinil may further increase the speed of onset of action of insulin formulated in compositions of the system of the invention. Suitably, the concentration of treprostinil in the composition 30 is in the range of 0.1-12 μg/ml e.g. 0.1-10 μg/ml, 0.1-9 μg/ml, 0.1-8 μg/ml, 0.1-7 μg/ml, 0.1-6 μg/ml, 0.1-5 μg/ml, 0.1-4 μg/ml, 0.1-3 μg/ml, 0.1-2 μg/ml, 0.5-2 μg/ml e.g. about 1 μg/ml.
In one embodiment, the composition does not contain a vasodilator. In a 35 further embodiment, the composition does not contain treprostinil, nicotinamide, nicotinic acid or a salt thereof.
Compositions of the system may optionally include other beneficial components including stabilising agents. For example, amino acids such as arginine 40
20
or proline may be included which may have stabilising properties. Thus, in one embodiment, the compositions of the system comprise arginine.
In an embodiment of the invention the compositions are free of acids selected from glutamic acid, ascorbic acid, succinic acid, aspartic acid, maleic acid, fumaric 5 acid, adipic acid and acetic acid and are also free from the corresponding ionic forms of these acids.
In an embodiment of the invention the compositions of the system are free of arginine. 10
In an embodiment of the invention the compositions of the system are free of protamine and protamine salts.
In an embodiment of the invention the compositions of the system are free of 15 magnesium ions.
The addition of magnesium ions e.g. in the form of magnesium chloride may provide a stabilising effect. Thus, in an embodiment of the invention the composition contains magnesium ions e.g. MgCl2. 20
In an embodiment of the invention the compositions of the system are free of calcium ions.
Compositions of the system may further comprise an additional 25 therapeutically active agent (an “active agent”), in particular an agent of use in the treatment of diabetes (i.e. in addition to the insulin compound in particular the rapid-acting insulin compound) e.g. an amylin analogue or a GLP-1 agonist. In one embodiment, the composition further comprises an amylin analogue such as pramlintide, suitably at a concentration of 0.1-10 mg/ml e.g. 0.2-6 mg/ml. In one 30 embodiment, the composition further comprises a GLP-1 agonist such as liraglutide, dulaglutide, albiglutide, exenatide or lixisenatide, suitably at a concentration of 10 μg/ml to 50 mg/ml e.g. 200 μg/ml to 10 mg/ml or 1 mg/ml to 10 mg/ml.
Suitably the compositions of the system are sufficiently stable that the 35 concentration of high molecular weight species remains low upon extended storage. The term “high molecular weight species” as used herein, refers to any irreversibly formed component of the protein content which has an apparent molecular weight at least about double the molecular weight of the parent insulin compound, as detected by a suitable analytical method, such as size-exclusion chromatography. That is, 40
21
high molecular weight species are multimeric aggregates of the parent insulin compound. The multimeric aggregates may comprise the parent protein molecules with considerably altered conformation or they may be an assembly of the parent protein units in the native or near-native conformation. The determination of high molecular weight species can be done using methods known in the art, including 5 size exclusion chromatography, electrophoresis, analytical ultracentrifugation, light scattering, dynamic light scattering, static light scattering and field flow fractionation.
Suitably the compositions of the system are sufficiently stable that they remain substantially free of visible particles after storage at 30°C for at least one 10 month or more, two months or more, or three months or more. Visible particles are suitably detected using the 2.9.20. European Pharmacopoeia Monograph (Particulate Contamination: Visible Particles). For example, a composition is substantially free of visible particles if it has a Visual score according to Visual Assessment Scoring Method B of 1, 2 or 3, especially 1 or 2 according to the 15 definition given in the Examples section.
Suitably the compositions of the system are sufficiently stable that there is minimal increase in soluble aggregates such as <0.5%, <0.2% or <0.1% increase after storage at 30°C for one month or more, two months or more or three months or 20 more. Soluble aggregates are suitable detected using SEC (see General Methods).
Suitably the compositions of the system are sufficiently stable that the concentration of related species remains low upon extended storage. The term “related species” as used herein, refers to any component of the protein content 25 formed by a chemical modification of the parent insulin compound, particularly desamido or cyclic imide forms of insulin. Related species are suitably detected by RP-HPLC.
In a preferred embodiment, the composition of the system of the invention 30 retains at least 95%, e.g. at least 96%, e.g. at least 97%, e.g. at least 98%, e.g. at least 99% parent insulin compound (by weight of total protein) after storage at 30°C for one, two or three months. The percentage of insulin compound (by weight of total protein) may be determined by size-exclusion chromatography or RP-HPLC.
35
In a preferred embodiment, the composition of the system of the invention comprises no more than 4% (by weight of total protein), preferably no more than 2% high molecular weight species (e.g. visible particles and/or soluble aggregates) after storage at 30°C for one, two or three months.
22
In a preferred embodiment, the composition of the system of the invention comprises no more than 4% (by weight of total protein), preferably no more than 2%, preferably no more than 1% A-21 desamido form of the insulin compound after storage at 30°C for one, two or three months.
5
In preferred embodiments, a composition of the system of the invention should exhibit an increase in high molecular weight species (e.g. visible particles and/or soluble aggregates) during storage which is at least 10% lower, preferably at least 25% lower, more preferably at least 50% lower, than a composition lacking the alkyl glycoside as non-ionic surfactant but otherwise identical, following storage 10 under the same conditions (e.g. 30°C) and length of time (e.g. one, two or three months).
In preferred embodiments, a composition of the system of the invention should exhibit an increase in related species during storage which is at least 10% 15 lower, preferably at least 25% lower, more preferably at least 50% lower, than a composition lacking the alkyl glycoside as non-ionic surfactant but otherwise identical, following storage under the same conditions (e.g. 30°C) and length of time (e.g. one, two or three months).
20
The speed of action of a composition of the system of the invention may be determined in the Diabetic Pig Pharmacokinetic/Pharmacodynamic Model (see Examples, General Methods (c)). In preferred embodiments, a composition of the present invention exhibits a Tmax (i.e. time to peak insulin concentration) that is at least 20% shorter, preferably at least 30% shorter than a composition lacking the 25 zinc binding species having a logK with respect to zinc ion binding in the range 4.5-12.3 (e.g. in the range 4.5-10) at 25 °C but otherwise identical, using the model. In preferred embodiments, a composition of the present invention exhibits an area under the curve on the pharmacodynamics profile within the first 45 minutes after injection that is at least 20% greater, preferably at least 30% greater than a 30 composition lacking the zinc binding species having a logK with respect to zinc ion binding in the range 4.5-12.3 (e.g. in the range 4.5-10) at 25 °C but otherwise identical, using the model.
In one embodiment, the composition of the system of the invention comprises 35 (i) insulin lispro at a concentration of 400-1000 U/ml e.g. 500-1000 U/ml, (ii) ionic zinc, (iii) optionally a zinc binding species at a concentration of 1 mM or more selected from species having a logK with respect to zinc ion binding in the range 4.5-12.3 at 25 °C e.g. citrate, and (iv) a non-ionic surfactant which is an alkyl glycoside; and wherein the composition is substantially free of EDTA and any other zinc binding 40
23
species having a logK with respect to zinc ion binding of more than 12.3 at 25 °C, which exhibits a Tmax (i.e. time to peak insulin concentration) that is at least 20% shorter, preferably at least 30% shorter than an aqueous composition consisting of: insulin lispro (100 U/ml), sodium phosphate (13.2 mM), glycerol (174 mM), m-cresol (29 mM), ionic zinc (19.7 μg/ml, excluding counter-ion) adjusted to pH 7.3, using the 5 Diabetic Pig Pharmacokinetic/Pharmacodynamic Model (see Examples, General Methods (c)). In another embodiment, the present invention provides a composition comprising (i) insulin lispro at a concentration of 400-1000 U/ml e.g. 400-1000 U/ml e.g. 500-1000 U/ml, (ii) ionic zinc, (iii) optionally a zinc binding species at a concentration of 1 mM or more selected from species having a logK with respect to 10 zinc ion binding in the range 4.5-12.3 at 25 °C e.g. citrate, and (iv) a non-ionic surfactant which is an alkyl glycoside; and wherein the composition is substantially free of EDTA and any other zinc binding species having a logK with respect to zinc ion binding of more than 12.3 at 25 °C, which exhibits an area under the curve on the pharmacodynamics profile within the first 45 minutes after injection that is at least 15 20% greater, preferably at least 30% greater than an aqueous composition consisting of: insulin lispro (100 U/ml), sodium phosphate (13.2 mM), glycerol (174 mM), m-cresol (29 mM), ionic zinc (19.7 μg/ml, excluding counter-ion) adjusted to pH 7.3, using the Diabetic Pig Pharmacokinetic/Pharmacodynamic Model (see Examples, General Methods (c)). 20
In one embodiment, the composition of the system of the invention comprises (i) insulin aspart at a concentration of 400-1000 U/ml e.g. 400-1000 U/ml e.g. 500-1000 U/ml, (ii) ionic zinc, (iii) optionally a zinc binding species at a concentration of 1 mM or more selected from species having a logK with respect to zinc ion binding in 25 the range 4.5-12.3 at 25 °C e.g. citrate, and (iv) a non-ionic surfactant which is an alkyl glycoside; and wherein the composition is substantially free of EDTA and any other zinc binding species having a logK with respect to zinc ion binding of more than 12.3 at 25 °C, which exhibits a Tmax (i.e. time to peak insulin concentration) that is at least 20% shorter, preferably at least 30% shorter than an aqueous composition 30 consisting of: insulin aspart (100 U/ml), sodium phosphate (7 mM), glycerol (174 mM), sodium chloride (10 mM), phenol (15.9 mM), m-cresol (15.9 mM) and ionic zinc (19.7 μg/ml, excluding counter-anion) adjusted to pH 7.4, using the Diabetic Pig Pharmacokinetic/Pharmacodynamic Model (see Examples, General Methods (c)).
35
In another embodiment, the present invention provides a composition comprising (i) insulin aspart at a concentration of 400-1000 U/ml e.g. 400-1000 U/ml e.g. 500-1000 U/ml, (ii) ionic zinc, (iii) optionally a zinc binding species at a concentration of 1 mM or more selected from species having a logK with respect to zinc ion binding in the range 4.5-12.3 at 25 °C e.g. citrate, and (iv) a non-ionic 40
24
surfactant which is an alkyl glycoside; and wherein the composition is substantially free of EDTA and any other zinc binding species having a logK with respect to zinc ion binding of more than 12.3 at 25 °C, which exhibits an area under the curve on the pharmacodynamics profile within the first 45 minutes after injection that is at least 20% greater, preferably at least 30% greater than an aqueous composition 5 consisting of: insulin aspart (100 U/ml), sodium phosphate (7 mM), glycerol (174 mM), sodium chloride (10 mM), phenol (15.9 mM), m-cresol (15.9 mM) and ionic zinc (19.7 μg/ml, excluding counter-anion) adjusted to pH 7.4, using the Diabetic Pig Pharmacokinetic/Pharmacodynamic Model (see Examples, General Methods (c)).
10
In preferred embodiments, a composition of the system of the invention is bioequivalent to a standard composition comprising the insulin compound at 100 U/ml.
As used herein, “bioequivalent” means that the composition of the system of 15 the invention has an equivalent or similar pharmacokinetic/pharmacodynamic (PK/PD) profile to a standard composition. For example, the composition of the system of the invention exhibits a TMAX or T½MAX (measured in accordance with the Diabetic Pig Pharmacokinetic/Pharmacodynamic Model described in section (c) of General Methods) which is substantially the same as (e.g. within ±20% of, e.g. within 20 ±10% of) that of the standard composition. Bioequivalence can also be established by applying the Student’s t-test to the pharmacokinetic/pharmacodynamics results achieved using two different compositions as described in the diabetic pig pharmacokinetic/pharmacodynamic model described in section (c) of General Methods. 25
By “standard composition” is meant a commercially available composition of the same insulin compound at a concentration of 100 U/ml such as HUMALOG® (for insulin lispro) or NOVORAPID® (for insulin aspart) or APIDRA® (for insulin glulisine).
30
In one embodiment, the composition of the system of the invention comprises an insulin compound at a concentration of 400-1000 U/mL e.g. 500-1000 U/mL and wherein the composition is bioequivalent to a standard composition comprising the insulin compound at a concentration of 100 U/mL. In another embodiment, the absorption of insulin compound into the blood stream of the mammal after 35 administration using the system is bioequivalent to a standard composition at a concentration comprising the insulin compound at a concentration of 100 U/mL. In another embodiment, the glucose reduction response caused by administration of a given amount of insulin compound to the mammal using the system is bioequivalent
25
to a standard composition comprising the insulin compound at a concentration of 100 U/mL.
In one embodiment, a composition of the system of the invention wherein the insulin compound is insulin lispro is bioequivalent to a commercial composition of 5 insulin lispro at a concentration of 100 U/ml e.g. an aqueous composition consisting of: insulin lispro (100 U/ml), sodium phosphate (13.2 mM), glycerol (174 mM), m-cresol (29 mM), ionic zinc (19.7 μg/ml, excluding counter-ion) adjusted to pH 7.3 (i.e. the composition of HUMALOG®).
10
In one embodiment, a composition of the system of the invention wherein the insulin compound is insulin aspart is bioequivalent to a commercial composition of insulin aspart at a concentration of 100 U/ml e.g. an aqueous composition consisting of: insulin aspart (100 U/ml), sodium phosphate (7 mM), glycerol (174 mM), sodium chloride (10 mM), phenol (15.9 mM), m-cresol (15.9 mM) and ionic zinc (19.7 μg/ml, 15 excluding counter-anion) adjusted to pH 7.4 (i.e. the composition of NOVORAPID®).
According to further aspects of the invention, there is provided a composition of the system of the invention for use in the treatment of a subject suffering from diabetes mellitus. There is also provided a method of treatment of diabetes mellitus 20 which comprises administering to a subject in need thereof an effective amount of a composition of the system of the invention.
A typical insulin dose of the composition of the system of the invention is 2-100U e.g. 2-30 U, e.g. 5-15 U. Administration should suitably occur in the window 25 between 15 minutes before eating (i.e. before start of a meal) and 15 minutes after eating (i.e. after end of a meal).
In one embodiment, the composition of the system of the invention is co-administered with a long acting insulin such as insulin glargine or insulin degludec, 30 suitably at a concentration of 50-1000 U/ml e.g. 100-500 U/ml or 100-200 U/ml.
The composition of the system of the invention is for administration by injection, preferably by subcutaneous injection.
35
The system may comprise a dial mechanism enabling selection of a specific desired volume of the composition for delivery to the mammal. In one embodiment, the volume of composition selected for delivery is between 0.1-100 μL e.g. 0.25-50 μL, e.g. 0.50-20 μL. Suitably, the selected volume is determined by the dial mechanism in increments of 0.1-10 μL e.g. 0.25-5 μL e.g. 0.5-2 μL. Preferably, the 40
26
ratio between the delivered dose of insulin compound delivered (U) and the delivered volume (μL) is at least 0.4:1 e.g. at least 0.5: 1 e.g. at least 0.6:1.
The reservoir of the system which comprises the aqueous liquid pharmaceutical composition for delivery by means of said injection pen will typically 5 have a total volume of up to 3 mL e.g. 3 mL, e.g. 2 mL, e.g. 1 mL
The reservoir of the system will be retained in a container e.g. a cartridge. The containers may be a replaceable or refillable component of the system.
10
Injection pen systems provide a demanding environment for preserving the activity of insulin. For example, the reservoirs of such systems are exposed to warmth (particularly if carried close the body) and agitation (due to movement of the body).
15
Suitably the compositions of the system are sufficiently stable that they remain substantially free of visible particles after use for 4 weeks or more, 8 weeks or more, or 12 weeks or more. Visible particles are suitably detected using the 2.9.20. European Pharmacopoeia Monograph (Particulate Contamination: Visible Particles) combined with Scoring Method B (see General Methods). For example, a 20 composition is substantially free of visible particles if it has a Visual score according to Visual Assessment Scoring Method B of 1, 2 or 3, especially 1 or 2 according to the definition given in the Examples section.
Suitably the compositions of the system are sufficiently stable that they show 25 a minimal increase in soluble aggregates, such as less than 1% e.g. less than 0.5% e.g. less than 0.2%, following the use of the system of the invention for 4 weeks or more, 8 weeks or more, or 12 weeks or more. Soluble aggregates are suitably detected using Size Exclusion Chromatography (see General Methods).
30
In an embodiment, a composition of the system of the invention is more stable than in the absence of alkyl glycoside during operation of the injection pen for 4 weeks or more e.g. 8 weeks or more e.g. 12 weeks or more. For example, a composition of the system of the invention forms fewer visible particles and/or soluble aggregates than an identical composition in the absence of alkyl glucoside 35 during operation of the injection pen for 4 weeks or more e.g. 8 weeks or more, e.g. 12 weeks or more. Visual particles and soluble aggregates can be determined by Visual Assessment Scoring Method B and SEC (see General Methods).
27
In an embodiment, said in-use stability is indicated by the presence of fewer visible particles and/or soluble aggregates in the reservoir after use for the said number of weeks. In an embodiment, said in-use stability is indicated by the presence of fewer visible particles and/or soluble aggregates in a delivered dose after use for the said number of weeks. Visual particles and soluble aggregates can 5 be determined by Visual Assessment Scoring Method B and SEC (see General Methods).
In an embodiment, the injection pen of the system of the invention is disposable. Suitably, the injection pen is to be disposed of either when the insulin is 10 used up or after a specified time, such as after 2 weeks of use e.g. after 4 weeks of use, e.g. after 8 weeks of use.
In another embodiment, the injection pen is reusable and the reservoir is replaced as needed. Suitably, the reservoir is to be disposed of either when the 15 insulin is used up or after a specified time, such as after 2 weeks of use e.g. after 4 weeks of use, e.g. after 8 weeks of use.
The reservoir (often called “cartridge”) is typically in the shape of a syringe and the pen comprises a piston mechanism designed to apply the appropriate 20 pressure to dispense the required amount of insulin based on the values set (dialled) by the user.
Suitably the injector mechanism comprises a needle. The needle gauge typically ranges from 29 to 32 and the needle length typically ranges from 5 to 12 25 mm. In one embodiment, a new needle needs to be attached to the pen prior to each injection, using a specific mechanism such as screw thread or a push-on thread, and disposed of after use. In one embodiment the needle is a retractable needle. In one embodiment the injector mechanism comprises a spring-loaded retractable needle. The needle can be equipped with a protective shield to reduce the risk of needle-30 stick injuries or to allay patient anxiety about the needle use. Examples of commercially available insulin pens include KWIKPEN®, SOLOSTAR®, FLEXPEN®, FLEXTOUCH®, HUMAPEN®, NOVOPEN®.
In an embodiment, the system administers the composition subcutaneously 35 to the mammal. In an aspect of the invention, there is provided use of the system in the treatment of diabetes mellitus in said mammal. In an embodiment, the mammal requires 200 U of insulin per day or more. In another embodiment, the mammal has developed insulin resistance. In an embodiment, the mammal is a human.
28
In another embodiment, there is provided a method of treatment of diabetes mellitus which comprises administering to a mammal in need thereof an effective amount of an insulin compound containing composition via an injection pen using the system of the invention. In an embodiment, the mammal requires 200 U of insulin per day or more. In another embodiment, the mammal has developed insulin 5 resistance. Suitably, the mammal is a human.
Compositions of the system of the invention may be prepared by mixing the ingredients. For example, the insulin compound may be dissolved in an aqueous composition comprising the other components. Alternatively, the insulin compound 10 may be dissolved in a strong acid (typically HCl), after dissolution diluted with an aqueous composition comprising the other components, and then pH adjusted to the desired pH with addition of alkali (e.g. NaOH). As a variation on this method, a step of neutralising the acid solution may be performed before the dilution step and it may then not be necessary to adjust the pH after the dilution step (or a small adjustment 15 only may be necessary).
In another aspect of the invention, there is provided the use of an alkyl glycoside as a non-ionic surfactant to improve the stability of an insulin compound in an aqueous liquid pharmaceutical composition in an injection pen system comprising 20 an injection pen and an aqueous composition for delivery by means of said injection pen to a mammal, wherein the composition comprises (i) an insulin compound, (ii) ionic zinc and (iii) an alkyl glycoside as a non-ionic surfactant.
In a further aspect of the invention, there is provided a method of improving 25 the stability of an insulin compound to be administered by an injection pen system, which comprises adding an alkyl glycoside to an aqueous liquid pharmaceutical composition comprising the insulin compound and ionic zinc.
Systems of the invention in at least some embodiments are expected to have 30 one or more of the following advantageous properties:
• The systems can deliver high strength insulin that is rapid acting or ultra-rapid acting;
• The systems can deliver larger quantity of insulin within the in-use period, which improves convenience for the user, particularly if the 35 user has developed insulin resistance or requires large quantities of insulin for a different reason;
• The systems can be used for extended periods of time, such as >4 weeks;
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• Compositions of the system have good physical stability during use, for example after use for a number of weeks or months;
• Compositions of the system have good physical stability upon storage, especially as measured by the amount of HMWS or visual detection of particles; 5
• Compositions of the system have good chemical stability upon storage, especially as measured by the amount of related products e.g. products of deamidation;
• Compositions of the system have rapid speed of action, typically faster than normal human insulin, upon administration to a subject; 10
• Compositions of the system have rapid speed of action, typically as fast as a standard composition with insulin compound concentration of 100 U/ml;
• Compositions of the system have high insulin concentration while maintaining a rapid speed of action. 15
ABBREVIATIONS
DETA diethylenetriamine
EDTA ethylenediaminetetraacetate
EGTA ethyleneglycoltetraacetate 20
HPLC high performance liquid chromatography
HMWS high molecular weight species
RP reverse phase
SEC size-exclusion chromatography
TETA triethylenetetramine 25
PD pharmacodynamic
PK pharmacokinetic
EXAMPLES
General Methods 30
(a) Size exclusion chromatography (SEC)
Ultra-high performance size exclusion chromatography of insulin preparations was performed using the Waters ACQUITY H-class Bio UPLC® system with a 1.7 μm Ethylene Bridged Hybrid 125 Å pore packing material in a 300 mm by 4.6 mm column. The column was equilibrated in 0.65 mg/ml L-arginine, 20% v/v acetonitrile, 35 15%v/v glacial acetic acid mobile phase and 10 μl of sample, acidified with 0.01M HCl, was analysed at 0.4 mL/min, with 276 nm UV detection. All analyses were performed at ambient temperature.
40
30
(b) Reversed-phase chromatography (RP-HPLC)
Ultra-high performance reverse phase chromatography was performed using the Waters ACQUITY H-class Bio UPLC® system with a 1.7 μm Ethylene Bridged Hybrid particle, 130 Å pore resin trifunctionally immobilised with a C18 ligand in a 50 mm by 2.1 mm column. Insulin samples were bound in a 82%w/v Na2SO4, 18% v/v 5 acetonitrile, pH 2.3 mobile phase and eluted in 50% w/v Na2SO4, 50% v/v acetonitrile gradient flow. 2 μl of sample was acidified with 0.01M HCl and analysed at 0.61 mL/min, with 214 nm UV detection. All analyses were performed at 40°C.
(c) The Diabetic Pig Pharmacokinetic/Pharmacodynamic Model: Method for determining speed of action 10
10 male diabetic Yucatan miniature pigs were used. Pigs were injected subcutaneously with a sample of the test formulation and blood was taken (1 or 2 ml) at various time-points (min) with respect to the injection up to around 240 min after the injection. For pharmacodynamics profile, serum was analysed for glucose (using a commercially available glucometer). For pharmacokinetic profile, insulin 15 concentration was determined in the serum using an immunoassay.
In order to evaluate the formulations for bioequivalence, mean values of TMAX (i.e. time to reach the maximum insulin concentration in serum) and corresponding standard deviation were calculated across the whole set of 10 pigs used in the study. Similarly, mean values of T½MAX (i.e. time to reach half of the maximum 20 concentration) and corresponding standard deviation were calculated across the whole set of 10 pigs used in the study. Student t-test (95% confidence interval) was subsequently applied to allow assessment of bioequivalence between any two formulations tested. If the p-value of the t-test applied to the results populations of two samples was ≥0.05 the samples were considered bioequivalent, if the result was 25 <0.05 then the samples were considered non-bioequivalent.
(d) Visual assessment
Visible particles are suitably detected using the 2.9.20. European Pharmacopoeia Monograph (Particulate Contamination: Visible Particles). The apparatus required consists of a viewing station comprising: 30
• a matt black panel of appropriate size held in a vertical position
• a non-glare white panel of appropriate size held in a vertical position next to the black panel
• an adjustable lampholder fitted with a suitable, shaded, white-light source and with a suitable light diffuser (a viewing illuminator containing two 13 W 35 fluorescent tubes, each 525 mm in length, is suitable). The intensity of illumination at the viewing point is maintained between 2000 lux and 3750 lux.
Any adherent labels are removed from the container and the outside washed and dried. The container is gently swirled or inverted, ensuring that air bubbles are not 40
31
introduced, and observed for about 5 s in front of the white panel. The procedure is repeated in front of the black panel. The presence of any particles is recorded.
The visual scores are ranked as follows:
Visual Assessment Scoring Method A 5
Visual score 1: clear solution free of visible particles
Visual score 2: slight particle formation
Visual score 3: more significant precipitation
Visual Assessment Scoring Method B
Visual score 1: Clear solution, virtually free of particles 10
Visual score 2: ~ 5 very small particles
Visual score 3: ~10-20 very small particles
Visual score 4: 20-50 particles, including larger particles
Visual score 5: >50 particles, including larger particles
Whilst the particles in samples with visual scores 4 and 5 are clearly detectable on 15 casual visual assessment under normal light, samples with visual score 1-3 generally appear as clear solutions on the same assessment. Samples with visual scores 1-3 are considered to be “Pass”; samples with visual score 4-5 are considered to be “Fail”.
20
Example 1 – Example formulations
The following example formulations may be prepared:
Example A:
Insulin lispro 100 U/ml 25
We claim:
1. An injection pen system comprising an injector mechanism and a reservoir comprising an aqueous liquid pharmaceutical composition for delivery by means of said injector mechanism to a mammal wherein the composition 5 comprises (i) an insulin compound, (ii) ionic zinc and (iii) an alkyl glycoside as a non-ionic surfactant.
2. A system according to claim 1, wherein the insulin compound is not insulin glargine.
3. A system according to claim 1, wherein the insulin compound is insulin lispro. 10
4. A system according to claim 1, wherein the insulin compound is insulin aspart.
5. A system according to claim 1, wherein the insulin compound is insulin glulisine.
6. A system according to claim 1, wherein the insulin compound is recombinant 15 human insulin.
7. A system according to claim 1, wherein the insulin compound is not recombinant human insulin.
8. The system according to any one of claims 1 to 6, wherein the insulin compound is present at a concentration of 10-1000 U/ml. 20
9. The system according to claim 8, wherein the insulin compound is present at a concentration of 50-1000 U/ml.
10. The system according to claim 8, wherein the insulin compound is present at a concentration of 10-250 U/ml.
11. The system according to claim 9, wherein the insulin compound is present at 25 a concentration of 400-1000 U/ml.
12. The system according to claim 11, wherein the insulin compound is present at a concentration of 400-1000 U/ml e.g. 500-1000 U/ml, e.g. 600-1000 U/ml, e.g. 700-1000 U/ml, e.g. 800-1000 U/ml, e.g. 900-1000 U/ml, e.g. 1000 U/ml.
13. The system according to any one of claims 1 to 12, wherein the ionic zinc is 30 present at a concentration of more than 0.05% by weight of zinc based on the weight of insulin compound in the composition.
14. The system according to claim 13, wherein the ionic zinc is present at a concentration of more than 0.5% by weight of zinc based on the weight of insulin compound in the composition. 35
15. The system according to claim 14, wherein the ionic zinc is present at a concentration of 0.5-1% by weight of zinc based on the weight of insulin compound in the composition.
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16. The system according to any one of claims 1 to 15, wherein the composition further comprises a zinc binding species at a concentration of 1 mM or more selected from species having a logK with respect to zinc ion binding in the range 4.5-12.3 at 25 °C.
17. The system according to any one of claims 1 to 16, wherein the composition 5 is substantially free of EDTA and any other zinc binding species having a logK with respect to zinc ion binding of more than 12.3 at 25 °C.
18. The system according to claim 16 or 17, wherein the zinc binding species is selected from citrate, pyrophosphate, aspartate, glutamate, cysteine, cystine, glutathione, ethylenediamine, histidine, DETA and TETA. 10
19. The system according to claim 18, wherein the zinc binding species is citrate.
20. The system according to claim 19, wherein the source of the citrate is citric acid.
21. The system according to any one of claims 16 to 20, wherein the zinc binding species having a logK with respect to zinc ion binding in the range 4.5-12.3 is 15 present at a concentration of 1-60 mM.
22. The system according to any one of claims 16 to 21, wherein the molar ratio of ionic zinc to zinc binding species is 1:3 to 1:175.
23. The system according to claim 16 or claim 17, wherein the zinc binding at a concentration of 1 mM or more selected from species having a logK with 20 respect to zinc ion binding in the range 4.5-10 at 25 °C.
24. The system according to claim 16 or claim 17, which is substantially free of zinc binding species having a logK with respect to zinc ion binding of 10-12.3 at 25 °C.
25. The system according to any one of claims 1 to 24, wherein the alkyl 25 glycoside is selected from the group consisting of dodecyl maltoside, dodecyl glucoside, octyl glucoside, octyl maltoside, decyl glucoside, decyl maltoside, decyl glucopyranoside, tridecyl glucoside, tridecyl maltoside, tetradecyl glucoside, tetradecyl maltoside, hexadecyl glucoside, hexadecyl maltoside, sucrose monooctanoate, sucrose monodecanoate, sucrose 30 monododecanoate, sucrose monotridecanoate, sucrose monotetradecanoate and sucrose monohexadecanoate.
26. The system according to claim 25, wherein the alkyl glycoside is dodecyl maltoside or decyl glucopyranoside.
27. The system according to claim 26, wherein the alkyl glycoside is dodecyl 35 maltoside.
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28. The system according to any one of claims 1 to 27, wherein the alkyl
glycoside is present at a concentration of 1-1000 μg/ml e.g. 5-500 μg/ml, 10-
200 μg/ml, 10-100 μg/ml or around 50 μg/ml.
29. The system according to claim 28, wherein the alkyl glycoside is present at a
5 concentration of 10-400 μg/ml e.g. 20-400 μg/ml, 50-400 μg/ml, 10-300
μg/ml, 20-300 μg/ml, 50-300 μg/ml, 10-200 μg/ml, 20-200 μg/ml, 50-200
μg/ml, 10-100 μg/ml, 20-100 μg/ml or 50-100 μg/ml.
30. The system according to any one of claims 1 to 29, wherein the composition
further comprises a tonicity modifying agent.
10 31. The system according to claim 30, wherein the tonicity modifying agent is an
uncharged tonicity modifying agent.
32. The system according to claim 31, wherein the uncharged tonicity modifying
agent is selected from the group consisting of trehalose, mannitol, glycerol
and 1,2-propanediol.
15 33. The system according to claim 32, wherein the uncharged tonicity modifying
agent is glycerol.
34. The system according to claim 30, wherein the tonicity modifying agent is a
charged tonicity modifying agent.
35. The system according to claim 34, wherein the charged tonicity modifying
20 agent is sodium chloride.
36. The system according to any one of claims 1 to 33, wherein the ionic strength
of the composition excluding any zinc binding species and the insulin
compound is <40 mM, e.g. <30 mM, <20 mM or <10 mM, wherein ionic
strength is calculated according to the formula I:
=
= 
n
X 1
2
x x 25 I 0.5 c z
in which cx is molar concentration of ion x (mol L-1), zx is the absolute value of
the charge of ion x and the sum covers all ions (n) present in the
composition, wherein the contribution of the insulin compound and zinc
binding species (if present) should be ignored for the purposes of the
30 calculation.
37. The system according to any one of claims 1 to 36, wherein the composition
is substantially isotonic.
38. The system according to any one of claims 1 to 37, wherein the pH of the
composition is in the range 5.5 to 9.0.
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39. The system according to claim 38, wherein the pH of the composition is in the range 7.0 to 7.5 e.g. 7.4.
40. The system according to claim 38, wherein the pH of the composition is in the range 7.6 to 8.0 e.g. 7.8.
41. A system according to claim 39 or claim 40, wherein the composition 5 comprises a phosphate buffer e.g. sodium phosphate.
42. The system according to any of claims 1 to 41, wherein the composition further comprises a preservative.
43. The system according to claim 42, wherein the preservative is selected from the group consisting of phenol, m-cresol, chlorocresol, benzyl alcohol, 10 propylparaben, methylparaben, benzalkonium chloride and benzethonium chloride.
44. The system according to any one of claims 1 to 43, wherein the composition further comprises nicotinamide.
45. The system according to any one of claims 1 to 44, wherein the composition 15 further comprises nicotinic acid or a salt thereof.
46. The system according to any one of claims 1 to 45, wherein the composition further comprises treprostinil or a salt thereof.
47. The system according to claim 1, wherein the composition comprises (i) an insulin compound at a concentration of 50-500 U/ml (ii) ionic zinc, (iii) 20 optionally citrate as a zinc binding species at a concentration of 1 mM or more, and (iv) a non-ionic surfactant which is an alkylglycoside; and wherein the composition is substantially free of EDTA and any other zinc binding species having a logK with respect to zinc ion binding of more than 12.3 at 25 °C. 25
48. The system according to claim 47, wherein the citrate is present in the composition at a concentration of 10-30 mM.
49. The system according to claim 1, wherein the composition comprises (i) an insulin compound at a concentration of 400-1000 U/ml e.g. 500-1000 U/ml (ii) ionic zinc, (iii) optionally citrate as a zinc binding species at a concentration of 30 1 mM or more, and (iv) a non-ionic surfactant which is an alkyl glycoside; and wherein the composition is substantially free of EDTA and any other zinc binding species having a logK with respect to zinc ion binding of more than 12.3 at 25 °C.
50. The system according to claim 49, wherein the citrate is present in the 35 composition at a concentration of 30-60 mM.
51. The system of claim 49 or claim 50, wherein the ionic strength of the composition is less than 40 mM calculated using Formula I.
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52. The system of any one of claims 49-51, wherein the composition comprises <10 mM chloride.
53. The system of any one of claims 49-52, wherein the composition comprises an uncharged tonicity modifying agent.
54. The system according to any one of claims 1 to 53, wherein the composition 5 comprises an insulin compound at a concentration of 400-1000 U/mL e.g. 500-1000 U/mL and wherein the composition is bioequivalent to a standard composition comprising the insulin compound at a concentration of 100 U/mL.
55. The system according to any one of claims 1 to 53, wherein the absorption of 10 insulin compound into the blood stream of the mammal after administration using the system is bioequivalent to a standard composition comprising the insulin compound at a concentration of 100 U/mL.
56. The system according to any one of claims 1 to 53, wherein the glucose reduction response caused by administration of a given amount of insulin 15 compound to the mammal using the system is bioequivalent to a standard composition comprising the insulin compound at a concentration of 100 U/mL.
57. The system according to any of claims 1 to 56, comprising a dial mechanism enabling selection of a specific desired volume of the composition for delivery 20 to the mammal.
58. The system according to claim 57, wherein the volume of composition selected for delivery is between 0.1-100 μL e.g. 0.25-50 μL, e.g. 0.50-20 μL.
59. The system according to claim 58, wherein the selected volume is determined by the dial mechanism in increments of 0.1-10 μL e.g. 0.25-5 μL, e.g. 0.5-2 25 μL.
60. The system according to any one of claims 1 to 59, wherein the reservoir has a total volume of up to 3 mL e.g. 3 mL, e.g. 2 mL, e.g. 1 mL.
61. The system according to any one of claims 1 to 60, wherein the ratio between the delivered dose of insulin compound delivered (U) and the delivered 30 volume (μL) is at least 0.4:1 e.g. at least 0.5: 1, e.g. at least 0.6:1.
62. The system according to any one of claims 1 to 61, wherein the composition is more stable (e.g. forms fewer visible particles and/or soluble aggregates) than an identical composition in the absence of alkyl glycoside during operation of the pen for 4 weeks or more, or8 weeks or more, or 12 weeks or more. 35
63. The system according to any one of claims 1 to 62, wherein the injection pen is disposable.
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64. The system according to claim 63, wherein the injection pen is to be disposed of after use for up to 4 weeks e.g. up to 8 weeks e.g. up to 12 weeks.
65. The system according to any one of claims 1 to 64, wherein the injection pen system is reusable and the reservoir is replaced as needed.
66. The system according to claim 65, wherein the reservoir is to be disposed of 5 after the use for up to 4 weeks e.g. up to 8 weeks e.g. up to 12 weeks.
67. The system according to any one of claims 1 to 66, wherein the injector mechanism comprises a retractable needle.
68. The system according to claim 67, wherein the injector mechanism comprises a spring loaded retractable needle. 10
69. The system for use according to any one of claims 1 to 68, wherein the system is used to administer the composition subcutaneously to the mammal.
70. The system according to any one of claims 1 to 69, for use in the treatment of diabetes mellitus in said mammal.
71. The system according to claim 70, wherein the mammal requires 200 U of 15 insulin per day or more.
72. The system according to claim 71, wherein the mammal has developed insulin resistance.
73. The system for use according to claim 72, wherein the mammal is a human.
74. A method of treatment of diabetes mellitus which comprises administering to a 20 mammal in need thereof an effective amount of an insulin compound containing composition via an injection pen using a system according to any one of claims 1 to 73.
75. The method according to claim 74, wherein the mammal requires 200 U of insulin per day or more. 25
76. The method according to claim 74 or claim 75, wherein the mammal has developed insulin resistance.
77. The method according to any one of claims 74 to 76, wherein the mammal is a human.
78. Use of an alkyl glycoside as a non-ionic surfactant to improve the stability of 30 an insulin compound in an aqueous composition in an injection pen system comprising an injection pen and an aqueous liquid pharmaceutical composition for delivery by means of said injection pen to a mammal wherein the composition comprises (i) an insulin compound, (ii) ionic zinc and (iii) an alkyl glycoside as a non-ionic surfactant. 35
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79. A method of improving the stability of an insulin compound to be administered
by an injection pen system, which comprises adding an alkyl glycoside to an
aqueous liquid pharmaceutical composition comprising the insulin compound
and ionic zinc.