FR2978918A1 - Composition, useful for treating diabetes, comprises basal insulin, and dextran substituted by carboxylate charge carrier radicals and hydrophobic radicals, where composition is in the form of an injectable aqueous solution - Google Patents

Abstract

Composition comprises at least one basal insulin having an isoelectric point of 5.8-8.5, and at least one dextran substituted by carboxylate charge carrier radicals and hydrophobic radicals (I), where the composition is in the form of an injectable aqueous solution, and the pH is 6-8. Composition comprises at least one basal insulin having an isoelectric point of 5.8-8.5, and at least one dextran substituted by carboxylate charge carrier radicals and hydrophobic radicals of formula (I), where the composition is in the form of an injectable aqueous solution, and the pH is 6-8. R : -OH, -(f-[A]-COOH) nor -(g-[B1]-k-[D1]) m; D1 : at least one alkyl chain containing at least 8C; n : 0.1-2; m : 0-0.5; q : 3-50; A : linear or branched 1-4C chain, where the point of attachment is linked to a glucoside unit by a function-f comprising ether, ester and carbamate functions; B1 : linear or branched 1-4C divalent group, where the point of attachment is linked to a glycoside unit by a function-g comprising ether, ester and carbamate functions, and -D1 radical by function-k comprising ether, ester and carbamate functions, and the -D1 radical is -X(-/-Y1) p; X : 1-12C divalent group comprising C, N or O, optionally having amino or carboxyl functions and/or derived from a mono- or diamino amino acid or a dialcohol, or mono- or diamino polyethylene glycol, where the free acid functions are in the form of alkali metal cation salts comprising sodium cation salts or potassium cation salts; Y1 : linear or cyclic 8-20C alkyl, alkylaryl or arylalkyl (optionally substituted by 1-3C alkyl); and p : >= 1. Provided that: When p is 1, if Y1 is 8-14C alkyl, then qx m is >= 2, if Y1 is 15C alkyl, then qx m is >= 2, and if Y1 is 16-20C alkyl, then qx m is >= 1; and when p is >= 2, if Y1 is 8-11C alkyl, then qx m is >= 2, and if Y1 is 12-16C alkyl, then qx m is >= 0.3. An independent claim is included for an unidose formulation comprising the composition at a pH of 7-7.8 and prandial insulin. [Image] ACTIVITY : Antidiabetic. MECHANISM OF ACTION : None given.

Description

PH 7 INJECTABLE SOLUTION COMPRISING AT LEAST ONE BASIC INSULIN OF WHICH THE PI IS BETWEEN 5.8 AND 8.5 [0001] The invention relates to insulin (s) injection therapies for treating diabetes. Insulin therapy, or diabetes therapy by insulin injection, has in recent years remarkable progress thanks to the development of new insulins providing a correction of blood glucose patients, which allows 10 to better simulate physiological activity of the pancreas. To cover his daily insulin requirements, a diabetic patient currently has, schematically, two types of insulins with complementary actions: prandial insulins (or so-called fast-acting insulins) and basal insulins (or so-called slow acting insulins). [0004] Prandial insulins allow rapid management (metabolization and / or storage) of glucose provided during meals and snacks. The patient should inject mealtime insulin before each food intake, ie approximately 2 to 3 injections per day. The most widely used prandial insulins are recombinant human insulin, NovoLog® (insulin aspart NOVO NORDISK), Humalog® (insulin lispro ELI LILLY) and Apidra® (insulin glulisine SANOFI-AVENTIS). [0005] Basal insulins ensure the maintenance of glycemic homeostasis of the patient, outside the periods of food intake. They act essentially to block the endogenous production of glucose (hepatic glucose). The daily dose of basal insulin is generally 40-50% of the total daily insulin requirement. Depending on the basal insulin used, this dose is given in 1 or 2 injections, regularly distributed during the day. The most commonly used basal insulins are Levemir (NOVO NORDISK insulin detemir) and Lantus® (insulin glargine from SANOFI-AVENTIS). [0006] It should be noted that NPH (NPH insulin for Neutral Protamine Hagedorn, Humulin NPH®, Insulatard®) is the oldest basal insulin. This formulation is the result of a precipitation of human insulin (anionic at neutral pH) by a cationic protein, protamine. These microcrystals are dispersed in an aqueous suspension and dissolve slowly after subcutaneous injection. This slow dissolution ensures prolonged release of insulin. However this release does not ensure a constant concentration of insulin over time. The release profile is bell-shaped and lasts only 12 to 16 hours. It is injected twice a day. This NPH basal insulin is much less effective than modern basal insulins, Levernir® and Lantus®. NPH is an intermediate-acting basal insulin. The principle of the NPH has evolved with the appearance of fast analog insulins to give products called "Premix" offering both a fast action and an intermediate action. NovoLog Mix® (NOVO NORDISK) and Humalog Mixe (ELI LILLY) are formulations comprising fast-acting insulin, NovoLog® and Humalog®, partially complexed with protamine. These formulations thus contain insulin microcrystals whose action is said to be intermediate and a part of insulin which remains soluble and whose action is rapid. These formulations offer the advantage of fast insulin but they also have the defect of NPH, ie. a limited duration of action between 12 and 16 hours and insulin released in "bell". However, these products allow the patient to inject at one time an intermediate-acting basal insulin with a fast-acting prandial insulin. But many patients are anxious to reduce their number of injections. The basal insulins currently marketed and currently in clinical development can be classified according to the technical solution that allows to obtain the prolonged action and to date two approaches are used. The first, that of insulin detemir is the binding to albumin in vivo. It is an analogue, soluble at pH 7, which comprises an amino acid side chain attached to position B29 which, in vivo, allows this insulin to associate with albumin. Its prolonged action is mainly due to this affinity for albumin after subcutaneous injection. However, its pharmacokinetic profile does not cover a day, which is why it is most often used in two injections per day. [00011] Other basal insulins soluble at pH 7, such as Degludec® are currently under development. Degludec® has a fatty acid side chain attached to insulin. The second insulin glargine, is the precipitation at physiological pH. It is an analogue of human insulin obtained by elongation of the C-terminal part of the B-chain of human insulin by two arginine residues, and by substitution of the asparagine residue A21, by a glycine residue (US 5,656,722). The addition of the two arginine residues was designed to adjust the pI (isoelectric point) of insulin glargine to physiological pH, and thus make this insulin analogue insoluble in a physiological medium. The substitution of A21 was designed to make insulin glargine stable at acidic pH and thus be formulated as injectable solution at acidic pH.

During subcutaneous injection, the passage of insulin glargine from an acid pH (pH 4-4.5) to a physiological pH (neutral pH) causes its precipitation under the skin. The slow redissolution of insulin glargine micro-particles ensures a slow and prolonged action. The hypoglycemic effect of insulin glargine is almost constant over a period of 24 hours which allows most patients to be limited to a single injection per day. [00015] Insulin glargine is considered today as the best basal insulin marketed. [00016] However, the necessarily acidic pH of the insulin glargine formulation is a real disadvantage. This acidic pH of the insulin glargine formulation sometimes causes patients to have injection pain. To solve this side effect, no one has so far sought to solubilize these basal insulins whose isoelectric point is between 5.8 and 8.5 of insulin glargine type at neutral pH while maintaining a difference of solubility between the in-vitro medium (the container) and the in-vivo medium (under the skin), regardless of the pH. Indeed, the very principle of basal insulins, whose isoelectric point is between 5.8 and 8.5, insulin glargine type, explained above namely that they are soluble at acidic pH and precipitate at Physiological pH diverts the person skilled in the art from any solution in which insulin glargine insulin would be solubilized at pH 6-8 while retaining its essential property which is to precipitate in a subcutaneous environment. The Applicant has also noted that this acidic pH of the formulations of the basal insulins, whose isoelectric point is between 5.8 and 8.5, insulin glargine type, prevents any pharmaceutical combination with other proteins and in particular prandial insulins because the latter are not stable at acidic pH. The impossibility of formulating a prandial insulin at acidic pH is due to the fact that a mealtime insulin undergoes, under these conditions, a secondary reaction of deamidation in the A21 position; which does not meet the requirement of the US Pharmacopoeia, namely less than 5% of secondary product after 4 weeks at 30 ° C. In addition, this acidic pH of the formulations of basal insulins, whose isoelectric point is between 5.8 and 8.5, insulin glargine type, even prevents any extemporaneous combination with prandial insulins at neutral pH. [00023] A recent clinical study, presented at the 69th Scientific Sessions of the American Diabetes Association, New Orleans, Louisiana, 5-9 June 2009, made it possible to verify this limitation of use of insulin glargine. A dose of insulin glargine and a dose of mealtime insulin (in this case, insulin lispro) were mixed just prior to injection (E. Cengiz et al., 2010; Diabetes care - 33 (5): 1009- 12). This experiment revealed a significant delay in the pharmacokinetic and pharmacodynamic profiles of mealtime insulin, which may lead to postprandial hyperglycaemia and nocturnal hypoglycaemia. This study confirms the incompatibility of insulin glargine with fast-acting insulins currently on the market.

Moreover, the instructions for use of Lantus®, the commercial product based on insulin glargine company SANOFI-AVENTIS, explicitly tells users not to make a mixture with a mealtime insulin solution, which whether because of the serious risk of altering the pharmacokinetics and pharmacodynamics of insulin glargine and / or mixed prandial insulin. [00025] However, from a therapeutic point of view, clinical studies made public at the 70th Annual Scientific Sessions of the American Diabetes Association (ADA) of 2010. abstract 2163-PO and abstract number 0001-LB, in particular those conducted by the company SANOFI-AVENTIS, have shown that treatments that combine Lantus®, insulin glargine, and mealtime insulin are much more effective than treatments based on products of the "Premix" type, Novolog Mix® or Huma 10g Mix® . Regarding the combinations of insulin glargine and fast insulin, the company Biodel, described in particular in the patent application US 7,718,609 compositions comprising a basal insulin and prandial insulin at a pH of between 3.0 and 4.2 in the presence of a chelating agent and polyacids. This patent teaches how to make acid-prandial insulin compatible with insulin glargine. He does not teach how to prepare a combination of insulin glargine insulin and prandial insulin at neutral pH. From the analysis of the compositions described in the literature and patents it appears that insolubility at pH 7 basal insulin insulin glargine type is a prerequisite for its slow action. As a result, all the solutions proposed for combining them with other products, such as prandial insulins, are based on tests for solubilization or stabilization of prandial insulins at acidic pH, see for example WO 2007/121256 and WO 2009/021955. The present invention, by solving this problem allows: to propose an injectable composition for the treatment of diabetes, comprising a basal insulin whose isoelectric point is between 5.8 and 8.5, in the form of a homogeneous solution at pH 7, while maintaining its biological activity and its action profile; to propose a composition in the form of a formulation comprising a combination of a basal insulin whose isoelectric point is between 5.8 and 8.5 and of a mealtime insulin without modifying the activity profile of the prandial insulin soluble at pH 6-8 and unstable at acidic pH while maintaining the basal insulin-specific basal action profile; patients to reduce the number of their injections; said compositions to comply with the requirements of the American and European Pharmacopoeias. Surprisingly the compositions according to the invention allow maintenance of the duration of the hypoglycemic activity of basal insulin whose isoelectric point is between 5.8 and 8.5 despite its solubilization at pH 7 before injection . This remarkable property comes from the fact that insulin glargine insulin solubilized at pH 7 in the composition of the invention precipitates subcutaneously by a change in composition of the medium. The element triggering the precipitation of insulin-like insulin glargine is no longer the change in pH but a change in the composition of the environment during the passage of the pharmaceutical composition to the physiological medium. [00030] Surprisingly, the means found for solubilizing insulin glargine insulin at pH 7 fully preserves its biological activity. This maintenance observed in vivo can also be found in vitro by the preservation of the tertiary and quaternary structure of this basal insulin. For example, insulin glargine insulin at a concentration of 100 IU / ml is maintained in its hexameric form. In the combinations, this composition also makes it possible to preserve the tertiary and quaternary structure of insulin glargine and other prandial insulins. [00031] Surprisingly, in the combinations of insulin-like insulin glargine with mealtime insulin, objects of the invention, the rapid action of prandial insulin is preserved despite the precipitation of insulin-like insulin. insulin glargine. The invention relates to a composition in the form of an injectable aqueous solution, the pH of which is between 6.0 and 8.0, comprising at least: a) a basal insulin whose isoelectric point p1 is included between 5.8 and 8.5; b) a dextran substituted with radicals carrying carboxylate charges and hydrophobic radicals of formula I: ## STR2 ## in which: - R is -Ol-1 or chosen from the group consisting of the radicals: o - (f [A] -000H) no - (g- [B] -k- [D]), D having at least one alkyl chain having at least 8 carbon atoms, - n represents the degree of substitution of the glucosidic units by -f- [ A] -COOH and 0.1 S n S 2; m represents the degree of substitution of the glucoside units by -G- [B] -k- [D] and 0 <m 0.5; q represents the degree of polymerisation in glucosidic units, that is to say the average number of glucosidic units per polysaccharide chain; and 3 q - - (f- [AJ-COOH) no -A- is a linear radical or branched having 1 to 4 carbon atoms; said radical -A-: o being bound to a glucosidic unit by a function f selected from the group consisting of ether, ester and carbamate functions. - (g- [B] -k- [DJ] m, o -B- is an at least divalent linear or branched radical comprising 1 to 4 carbon atoms; said radical -B- o being bound to a glucosidic unit by a function g selected from the group consisting of ether, ester and carbamate functions; o being bound to a radical -D by a function k; k selected from the group consisting of ester, amide, and carbamate; said radical - D: - being a radical -X (-1-Y) p, X being an at least divalent radical comprising from 1 to 12 atoms selected from the group consisting of C, N or O atoms, optionally carrying carboxyl or amine functions and / or derived from an amino acid, a dihydric alcohol, a diamine or a mono- or polyethylene glycol mono- or diamine; Y being a linear or cyclic alkyl group, a C8 to C20 alkylaryl or arylalkyl, optionally substituted by one or more C1 to C3 alkyl groups; p? 1 and 1 a function selected from the group consisting of ester, amide and carbamate functions. f, g and k being identical or different. the acidic fiber functions being in the form of salts of alkaline cations chosen from the group consisting of Na and K +. and when p = 1, if Y is a C8 to C14 alkyl, then q * m? 2, if Y is C15 alkyl, then q * m? 2; and if Y is C16-C20 alkyl, then q * m? 1 - and when p 2, if Y is C8-C11 alkyl, then q * m? 2 and, if Y is C12-C16 alkyl, then q * m? 0.3.

In one embodiment, the radical - (f- [A] -COOH) 'is such that 25 - -A- is a radical comprising 1 carbon atom; said radical -A- being linked to a glucoside unit by an ether function.

In one embodiment, the radical - (g- [B] -k- [D]) is such that - -B- is a radical comprising 1 carbon atom; said radical -B- being linked to a glucoside unit by a g ether function, and, - X is a radical derived from an amino acid.

In one embodiment, the radical - (f [A] -COOH) E is such that - -A- is a radical comprising 1 carbon atom; said radical -A- being linked to a glucosidic unit by a function of ether, and - the radical - (g- [B] -k- [D]), is such that: - -B- is a radical comprising 1 carbon atom; said radical -B- being linked to a glucoside unit by a g ether function, and, - X is a radical derived from an amino acid. k is an amide function.

The structure drawn corresponds to the representation commonly used to represent dextran which is a polysaccharide consisting mainly of sequences (1,6) between glucoside units which is the representation adopted. Dextran also contains chains (1,3) to about 5% in general which are not deliberately represented, but which are of course included in the scope of the invention. By basal insulin whose isoelectric point is between 5.8 and 8.5 insulin insulin at pH 7 and whose duration of action is between 8 and 24 hours or higher in the standard models of diabetes. These basal insulins whose isoelectric point is between 5.8 and 8.5 are recombinant insulins whose primary structure has been modified mainly by introduction of basic amino acids such as Arginine or Lysine. They are described for example in the following patents, patent applications or publications WO 2003/053339, WO 2004/096854, US 5,656,722 and US 6,100,376. In one embodiment, the basal insulin whose isoelectric point is between 5.8 and 8.5 is insulin glargine. In one embodiment, the compositions according to the invention comprise 100 IU / ml (ie approximately 3.6 mg / ml) of basal insulin whose isoelectric point is between 5.8 and 8.5. In one embodiment, the compositions according to the invention comprise 40 IU / ml of basal insulin whose isoelectric point is between 5.8 and 8.5. In one embodiment, the compositions according to the invention comprise 200 IU / ml of basal insulin whose isoelectric point is between 5.8 and 8.5. In one embodiment, the mass ratio between the basal insulin, whose isoelectric point is between 5.8 and 8.5, and the substituted dextran, ie substituted dextran / basal insulin, is between 0.5. and 5. [00044] In one embodiment, the mass ratio between the basal insulin, whose isoelectric point is between 5.8 and 8.5, and the substituted dextran, ie substituted dextran / basal insulin, is included. between 0.5 and 3. In one embodiment, the mass ratio between the basal insulin, whose isoelectric point is between 5.8 and 8.5, and the substituted dextran, is substituted dextran / basal insulin. is in the range of 1 to 3. [00046] In one embodiment, the polysaccharide concentration is between 5 and 20 mg / ml. In one embodiment, the concentration of polysaccharide is between 5 and 10 mg / ml. [00048] In one embodiment, the compositions according to the invention further comprise a mealtime insulin. The prandial insulins are soluble at pH 7. [00049] "Prandial insulin" is understood to mean so-called fast or "regular" insulin. The so-called fast prandial insulins are insulins that must meet the needs caused by ingestion of proteins and carbohydrates during a meal, they must act in less than 30 minutes. [00051] In one embodiment, the so-called "regular" prandial insulins are chosen from the group comprising Humulin® (human insulin) and Novolin® (human insulin). The so-called fast acting mimic insulins are insulins which are obtained by recombination and which are modified to reduce their time of action. In one embodiment, the so-called fast acting (fast acting) prandial insulins are chosen from the group comprising Humalog® (insulin lispro), Apidra® (insulin glulisine) and NovoLog® (insulin aspart). [00054] In one embodiment, the compositions according to the invention comprise a total of 100 IU / ml (or about 3.6 mg / ml) of insulin with a combination of mealtime insulin and basal insulin whose point isoelectric is between 5.8 and 8.5. In one embodiment, the compositions according to the invention comprise a total of 40 IU / ml of insulin with a combination of prandial insulin and basal insulin whose isoelectric point is between 5.8 and 8, 5. In one embodiment, the compositions according to the invention comprise a total of 200 IU / ml of insulin with a combination of mealtime insulin and basal insulin whose isoelectric point is between 5.8 and 8.5. . The proportions between the basal insulin whose isoelectric point is between 5.8 and 8.5 and the mealtime insulin are, for example, in IU / ml of 25/75, 30/70, 40/60, 50/50, 60/40, 70/30. However any other proportion can be realized. In one embodiment, the compositions according to the invention also comprise zinc salts at a concentration of between 0 and 1000 μM. In one embodiment, the compositions according to the invention also comprise zinc salts at a concentration of between 200 and 600 μM. [00060] In one embodiment, the compositions according to the invention further comprise zinc salts at a concentration of between 50 and 500 μM. In one embodiment, the compositions according to the invention comprise buffers chosen from the group comprising Tris and phosphates at concentrations of between 0 and 100 mM, preferably between 15 and 50 mM. [00062] In one embodiment, the compositions according to the invention further comprise preservatives. In one embodiment, the preservatives are selected from the group consisting of m-cresol and phenol alone or in admixture. [00064] In one embodiment, the concentration of the preservatives is between 10 and 40 mM. The compositions according to the invention may further comprise additives such as tonicity agents such as glycerine, NaCl, mannitol and glycine. [00066] The compositions according to the invention may further comprise pharmacopoeial additives such as surfactants, for example polysorbate. The compositions according to the invention may further comprise all the excipients according to the pharmacopoeia and compatible with the insulins used at the concentrations of use. [00068] In general, the pH of the compositions is adjusted by addition of 10% HCl solution or 10% NaOH solution. In one embodiment, 0.3. . In one embodiment, 0.75 n 1.5. In one embodiment, 0.9 5 n 1.2. In one embodiment, 0.01 5 m <0.5. In one embodiment, 0.02 s m s 0.4. In one embodiment, 0.03 5 m 0.3. In one embodiment, 0.05 <m <0.2. In one embodiment, in one embodiment, in one embodiment, the radical - (f- [A] -COOH ) n is selected from the group consisting of the following sequences, f having the meaning given above:

~~ pH fOH 20

In one embodiment, the radical - (g- [B] -k- [D]) m is chosen from the group consisting of the following sequences; g, k and D having the meanings given above: [00080] In one embodiment, D is such that the radical X is an at least divalent radical derived from an amino acid selected from the group consisting of glycine, phenylalanine, lysine and aspartic acid. [00081] The radicals derived from amino acids can be either levorotatory or dextrorotatory. In one embodiment, D is such that the radical X is an at least divalent radical derived from ethylene glycol. [00083] In one embodiment, D is such that the radical X is an at least divalent radical derived from ethylenediamine. [00084] In one embodiment, D is such that the radical X is an at least divalent radical derived from a polyethylene glycol chosen from the group consisting of diethylene glycol and triethylene glycol. In one embodiment, D is such that the radical X is an at least divalent radical derived from a mono- or polyethylene glycol amine chosen from the group consisting of ethanolamine, diethylene glycol amine and triethylene. glycol amine. [00086] In one embodiment, D is such that the radical X is an at least divalent radical derived from a mono- or polyethylene glycol diamine chosen from the group consisting of diethylene glycol diamine and triethylene glycol diamine. In one embodiment, D is such that the group Y is an alkyl group derived from a hydrophobic alcohol selected from the group consisting of octanol (caprylic alcohol), 3,7-dimethyloctan-1 -ol, decanol (decyl alcohol), dodecanol (lauryl alcohol), tetradecanol (meringylic alcohol) and hexadecanol (cetyl alcohol). In one embodiment, D is such that the group Y is an alkyl group derived from a hydrophobic acid selected from the group consisting of decanoic acid, dodecanoic acid, tetradecanoic acid and hexadecanoic acid. [00089] In one embodiment, D is such that the X radical is derived from glycine, p = 1, the Y group is derived from octanol and the function is an ester function. In one embodiment, D is such that the radical X is derived from glycine, p = 1, the Y group is derived from dodecanol and the function 1 is an ester function. In one embodiment, D is such that the radical X is derived from glycine, p = 1, the Y group is derived from hexadecanol and the function 1 is an ester function. In one embodiment, D is such that the radical X is derived from phenylalanine, p = 1, the Y group is derived from octanol and the function 1 is an ester function. In one embodiment, D is such that the radical X is derived from phenylalanine, p = 1, the group Y is derived from 3,7-dimethyloctan-1-ol and the function is an ester function. . In one embodiment, D is such that the radical X is derived from aspartic acid, p = 2, the Y groups are derived from octanol and the functions 1 are ester functions. In one embodiment, D is such that the radical X is derived from aspartic acid, p = 2, the Y groups are derived from decanol and the functions 1 are ester functions. In one embodiment, D is such that the radical X is derived from aspartic acid, p = 2, the Y groups are derived from dodecanol and the functions 1 are ester functions. In one embodiment, D is such that the radical X is derived from ethylenediamine, the group Y is derived from dodecanoic acid and the function is an amide function. In one embodiment, D is such that the radical X is derived from diethylene glycol amine, p = 1, the group Y is derived from dodecanoic acid and the function is an ester function. [00099] In one embodiment, D is such that the radical X is derived from triethylene glycol diamine, p = 1, the group Y is derived from dodecanoic acid and the function is an amide function. In one embodiment, D is such that the radical X is derived from triethylene glycol diamine, p = 1, the group Y is derived from hexadecanoic acid and the function / is an amide function. [000101] In one embodiment: ## STR5 ## is such that A is the radical -CH 2 - and f is an ether function, o - (g- [B] -k - [D]) m is such that g is an ether function, B is the radical -CH2-, k is an amide function and D is such that X is derived from glycine, l is an ester function and Y is from octanol; Q = 38, n = 0.9 and m = 0.2,

[000102] In one embodiment: - (f- [A] -COOH) 'is such that A is the -CH2- radical and f is an ether function; where g is an ether function, B is the radical -CH 2 - glycine, p = 1, I is an ester function and Y is derived from hexadecanol; o q = 19, n = 1.0 and m = 0.1.

[000103] In one embodiment; Wherein A is -CH 2 - and f is an ether function; o - (g- [B] -k- [D]) m is such that g is an ether function, B is the radical -CH2-, k is an amide function and D is such that X is derived from phenylalanine, 5 p = 1, 1 is an ester function and Y is derived from octanol; o q = 38, n = 1.1 and m = 0.2.

In one embodiment: - (f- [A] -COOH) 'is such that A is the radical -CH2- and f is an ether function; O - (g- [B] -k- [D]) rr, is such that g is an ether function, B is the radical -CH2-, k is an amide function and D is such that X is derived from phenylalanine, p = 1, 1 is an ester function and Y is derived from octanol; o q = 19, n = 1.1 and m = 0.2.

In one embodiment o - (f [A] -COOH) n is such that A is the radical -CH2- and f is an ether function; o - (g- [B] -k- [D]) m is such that g is an ether function, B is the radical -CH2-, k is an amide function and D is such that X is derived from phenylalanine, p = 1, I is an ester function and Y is derived from 3,7-dimethyloctan-1-ol; O = 38, n = 1.0 and m = 0.1.

In one embodiment: - (f- [A] -COOH) n is such that A is the radical -CH2- and f is an ether function; o - (g- [B] -k- [D]) n, is such that g is an ether function, B is the radical -CH2-, k is an amide function and D is such that X is derived from aspartic acid, p = 2.1, are ester functions and Y are derived from octanol; o q = 38, n = 1.05 and m = 0.05.

In one embodiment: - (f [A] -COOH) 'is such that A is the radical -CH2- and f is an ether function; o - (g- [B] -k- [D]) ,,, is such that g is an ether function, B is the radical -CH2-, k is an amide function and D is such that X is derived from aspartic acid, p = 2.1, are ester functions and Y are derived from decanol; o q = 38, n = 1.05 and m = 0.05.

In one embodiment: - (f- [A] -COOH) 'is such that A is the radical -CH2- and f is an ether function, o - (g- [B] -k- [D]) m is such that g is an ether function, B is the radical -CH2-, k is an amide function and D is such that X is derived from aspartic acid, p = 2, 1 are ester functions and Y are derived from dodecanol oq = 19, n = 1.05 and m = 0.05.

In one embodiment: - (f- [A] -COOH) 'is such that A is the radical -CH2- and f is an ether function; o - (g- [B] -k- [D]) m is such that g is an ether function, B is the radical -CH2-, k is an amide function and D is such that X is derived from ethylenediamine, p = 1, is an amide function and Y is derived from dodecanoic acid; o q = 38, n = 1.0 and m = 0.1.

[000110] In one embodiment: - (f- [A] -COOH) 'is such that A is the -CH2-CH2- radical and f is an ester function; o - (g- [B] -k- [D]) m is such that g is an ester function, B is the radical -CH2-CH2-, k is an amide function and D is such that X is derived from glycine, p = 1.1 is an ester function and Y is derived from dodecanol; o q = 38, n = 1.3 and m = 0.1.

In one embodiment: - (f- [A] -COOH) 'is such that A is the radical -CH2- and f is a carbamate function; 0 - (g- [B] -k- [D]) m is such that g is a carbamate function, B is the radical -CH2-, k is an amide function and D is such that X is derived from aspartic acid, p = 2.1, are ester functions and Y are derived from octanol; o q = 38, n = 1.3 and m = 0.1. [000112] In one embodiment: - (f [A] -COOH) 'is such that A is the -CH2- radical and f is an ether function; o - (g- [B] -k- [D]), m is such that g is an ether function, B is the radical -CH2-, k is an amide function and D is such that X is derived from aspartic acid, p = 2, I are ester functions and Y are derived from dodecanol; o q = 4, n = 0.96 and m = 0.07.

In one embodiment: - (f- [A] -COOH) 'is such that A is the radical -CH2- and f is an ether function; o - (g- [B] -k- [D]) ,,, is such that g is an ether function, B is the radical -CH2-, k is an amide function and D is such that X is derived from diethylene glycol amine, p = 1, 1 is an ester function and Y is derived from dodecanoic acid; Where q = 38, n = 1.0 and m = 0.1.

In one embodiment: - (f- [A] -COOH), is such that A is the radical -CH2- and f is an ether function; O - (g- [B] -k- [D]), is such that g is an ether function, B is the radical -CH2-, k is an amide function and D is such that X is from triethylene glycol diamine, p = 1, 1 is an amide function and Y is derived from dodecanoic acid; o q = 38, n = 1.0 and m = 0.1,

In one embodiment: - (f [A] -COOH) 'is such that A is -CH2- and f is an ether function; o - (g- [B] -k- [D]), r, is such that g is an ether function, B is the radical -CH2-, k is an amide function and D is such that X is derived triethylene glycol diamine, p = 1, I is an amide function and Y is derived from hexadecanoic acid; oq = 38, n = 1.05 and m = 0.05. In one embodiment: o - (f- [A] -COOH) n is such that A is the radical -CH2- and f is an ether function; o - (g- [B] -k- [D]), m is such that g is an ether function, B is the radical -CH2-, k is an amide function and D is such that X is derived from glycine, p = 1, is ester functional and Y is hexadecanol; o q = 19, n = 1.05 and m = 0.05.

In one embodiment: - (f- [A] -COOH) 'is such that A is the radical -CH2- and f is an ether function; o - (g- [B] -k- [D]) m is such that g is an ether function, B is the radical -CH2-, k is an amide function and D is such that X is derived from glycine, p = 1, is ester functional and Y is hexadecanol; o q = 38, n = 0.37 and m = 0.05.

[000118] The invention also relates to single-dose formulations having a pH of between 7 and 7.8, comprising a basal insulin whose isoelectric point is between 5.8 and 8.5 and a prandial insulin. [000119] In one embodiment, the basal insulin whose isoelectric point is between 5.8 and 8.5 is insulin glargine. [000120] In one embodiment, the mealtime insulin is selected from the group consisting of Humulin® (human insulin) and Novolin® (human insulin). In one embodiment, the prandial insulin is chosen from the group comprising Humalog® (insulin lispro), Apidra® (insulin glulisine) and NovoLog® (insulin aspart). The solubilization at pH between 7 and 7,8 of the basal insulins whose isoelectric point is between 5.8 and 8.5 by the polysaccharides of formula I can be ascertained and controlled in a simple way, at the naked eye, thanks to a change of appearance of the solution. Moreover, and just as importantly, the Applicant has been able to verify that a basal insulin whose isoelectric point is between 5.8 and 8.5, solubilized in the presence of a polysaccharide of formula I had lost none of his slow insulin action. The preparation of a composition according to the invention has the advantage of being able to be achieved by simply mixing an aqueous solution of basal insulin whose isoelectric point is between 5.8 and 8.5, a solution of meal insulin, and a polysaccharide of formula I, in aqueous solution or in freeze-dried form. If necessary, the pH of the preparation is adjusted to pH 7. [000125] Examples Part A Polysaccharides

Table 1 below shows, without limitation, examples of polysaccharides used in the compositions according to the invention. SUBSTITUTE POLYSACCHARIDES USUAL NAME α-COONa-gBkD Polysaccharide 1 O o Dextranemethylcarboxylate of q: 38 OO sodium modified with n 0.9 Oxytyl glycinate m: 0.2 0 ON ~~ OG o Polysaccharide 2 q 19 n . 1,0 0 m: 0.1 O Na Polysaccharide 16 Dextranethylcarboxylate C: 19 n: 1.05 HO sodium modified with m: 0.05 14 cetyl glycinate Polysaccharide 17 pq: 38 n: 0.37 m: 0 Polysaccharide 3 0 ~ / 0 Na O 6 Dextranmethylcarboxylate of q: 38 ON 0 sodium modified with octyl n: 1,0 y phenylalaninate m: 0,1 0 Polysaccharide 4 q: 19 n: 1,1 m: 0.2% Na 2 O-Polysaccharide 0.02% dimethylmethylcarboxylate 0.01% sodium modified with n-phenylalaninate 3.7: 0.1 dimethyl-1-octyl 0 ~ / 0 Na O Polysaccharide 6 0 q: 38 Aspartate Modified Dextranmethylcarboxylate m: 0.05 g dioctyl O 0 '0Na e Polysaccharide 7 OO Dextranemethylcarboxylate II q: 38' 0 ~ / sodium modified with aspartate n: 1.05 m: 0.05 0 `, didecyl ~ / s O e 0 Na O Polysaccharide 8 N_ p Dextranemethylcarboxylate q: 19 0 Off / sodium modified by aspartate n: 1.05 of dilauryl m: o, a5 0 ~ \ ~ o 0 0 O Na Polysaccharide 9 0 ~~ p Dextra N-methyl-2-amino-2-methyl-dodecaned amine N-methyl-carboxylate Modified: N-O-NH 2 O-N-O-Polysaccharide 10 0 0 0 Sodium Dextransuccinate q: 38 Modified by the glycinate of n: 1, 3 lauryl m: 0.1 Polysaccharide 11 0 oo No o N-methylcarboxylate of q: 1.3 H o sodium dextran carbamate m: 0.1 0 H modified by the aspartate of N o dioctyl H Polysaccharide 12 -0 ~ / 0 ~ \ / p Dextranmethylcarboxylate q: 4 e sodium modified by aspartate n: 0.96 p Na O dilauryl m: 0.07 OHN 0 p ~~ o Na Polysaccharide 13 b Dextranmethylcarboxylate of q: 38 oo sodium modified with 2- (2- n: 1.0 amino-ethoxy) ethyl m: 0.1 dodecanoate Polysaccharide 14 o opo ~ o o Dextranemethylcarboxylate of q: 38 o - sodium modified by the 2- (2- {2- n: 1.0 Na [dodecanoylamino] ethoxy} ethm: 0.1 o oxy) ethylamine o Polysaccharide 15 -'-o ~ / Na o1 ~ o Dextranemethylcarboxylate q: 38 δ - sodium modified with 2- (2- {2- n: 1,05 o [hexadecanoylamino] thoxy} m: 0.05 thoxy) ethylamine Table 1

Example A1: Preparation of Polysaccharide 1 [000127] 16 g (ie 296 mmol of hydroxyl) of dextran with a weight average molar mass of approximately 10 kg / mol (q = 38, PHARMACOSMOS) are solubilized in water at room temperature. 42 g / L. To this solution are added 30 ml of 10 N NaOH (296 mmol). The mixture is brought to 35 ° C and then 46 g (396 mmol) of sodium chloroacetate are added. The temperature of the reaction medium is brought to 60 ° C. at 0.5 ° C./min, then maintained at 60 ° C. for 100 minutes. The reaction medium is diluted with 200 ml of water, neutralized with acetic acid and purified by ultrafiltration on PES membrane 5 kDa against 6 volumes of water. The final solution is assayed by dry extract to determine the concentration of polysaccharide; then assayed by acid / base assay in water / acetone 50/50 (V / V) to determine the average number of methylcarboxylate units per glucosidic unit. According to the dry extract: [polysaccharide] = 31.5 mg / g [000129] According to the acid / base assay: the average number of methylcarboxylate units per glucoside unit is 1.05. The sodium dextranmethylcarboxylate solution is passed through a Purolite (anionic) resin to obtain the acidic dextran-methylcarboxylic acid which is then lyophilized for 18 hours. The octyl glycinate, para-toluenesulfonic acid salt is obtained according to the process described in US Patent 4,826,818. 10 g of dextranmethylcarboxylic acid (44.86 mmol methyl carboxylic acid) are solubilized in DMF at 60 g / L and then cooled to 0 ° C. 3.23 g of octyl glycinate, paratoluenesulfonic acid salt (8.97 mmol) is suspended in DMF at 100 g / l. 0.91 g of triethylamine (8.97 mmol) is then added to this suspension. Once the polysaccharide solution at 0 ° C., a solution of NMM (5.24 g, 51.8 mmol) in DMF (530 g / L) and 5.62 g (51.8 mmol) of EtOCOCI are then added. added. After 10 minutes of reaction, the octyl glycinate suspension is added. The medium is then maintained at 10 ° C. for 45 minutes. The medium is then heated to 30 ° C. A solution of imidazole (10.38 g in 17 ml of water) and 52 ml of water are added to the reaction medium. The polysaccharide solution is ultrafiltered on a 10 kDa PES membrane against 15 volumes of 0.9% NaCl solution and 5 volumes of water. The concentration of the polysaccharide solution is determined by dry extract. A fraction of solution is lyophilized and analyzed by 11-I NMR in D20 to determine the degree of substitution of methylcarboxylate octyl glycinate per glucosidic unit. [000133] According to the dry extract: [Polysaccharide 1] = 36.4 mg / g [000134] According to the acid / base assay: n = 0.9 [000135] According to the 1H NMR: m = 0.2.

Example A2 Preparation of Polysaccharide 2 [000136] Cetyl glycinate, a para-toluenesulfonic acid salt, is obtained according to the process described in US Pat. No. 4,826,818. By a method similar to that described in Example A1, a sodium dextranemethylcarboxylate, synthesized according to the process described in Ex. Al using a dextran with a weight average molecular weight of about 5 kg / mol (g). = 19, PHARMACOSMOS), modified with cetyl glycinate is obtained. According to the dry extract: [Polysaccharide 2] = 15.1 mg / g [000139] According to the acid / base assay: n = 1.05 [000140] According to the 1H NMR: m = 0.05.

Example A3: Preparation of Polysaccharide 3 [000141] Octyl phenylalaninate, para-toluenesulfonic acid salt is obtained according to the process described in US Pat. No. 4,826,818. By a method similar to that described in Example A1, a sodium dextranemethylcarboxylate, synthesized according to the process described in Example A1 using a dextran with a weight average molecular weight of about 10 kg / mol (g). = 38, PHARMACOSMOS), modified with octyl phenylalaninate is obtained. [000143] From the dry extract: [Polysaccharide 3] = 27.4 mg / g [000144] According to the acid / base assay: n = 1.0 [000145] According to 1 H NMR: m = 0.1.

Example A4: Preparation of Polysaccharide 4 [000146] By a process similar to that described in Example A3, a sodium dextranemethylcarboxylate, synthesized according to the method described in Example A1 using a dextran of average molecular weight by weight of about 5 kg / mol (q = 19, PHARMACOSMOS), modified with octyl phenylalaninate is obtained. [000147] From the dry extract: [Polysaccharide 4] = 21.8 mg / g [000148] According to the acid / base assay: n = 1.0 [000149] According to 1 H NMR: m = 0.1. Example A5: Preparation of Polysaccharide [000150] 3,7-Dimethyl-1-octyl phenylalaninate, para-toluenesulfonic acid salt is obtained according to the process described in US Pat. No. 4,826,818. By a method similar to that described in Example A1, a sodium dextranemethylcarboxylate, synthesized according to the process described in Example A1 using a dextran with a weight average molecular weight of about 10 kg / mol (q). = 38, PHARMACOSMOS), modified with 3,7-dimethyl-1-octyl phenylalaninate is obtained. [000152] From the dry extract: [Polysaccharide 5] = 24.3 mg / g [000153] According to the acid / base assay: n = 1.0 [000154] According to the "H: m = 0.1 Example A6: Preparation of Polysaccharide 6 [000155] Dioctyl aspartate, para-toluenesulphonic acid salt, is obtained according to the process described in US Patent 4,826,818. [000156] By a process similar to that described in US Pat. Example A1, a sodium dextranemethylcarboxylate, synthesized according to the method described in Example AI using a dextran of weight average molecular weight of about 10 kg / mol (q = 38, PHARMACOSMOS), modified by aspartate of dioctyl is obtained. [000157] From the dry extract: [Polysaccharide 6] = 22.2 mg / g [000158] From the acid / base assay: n = 1.05 [000159] Based on NMR H: m = 0.05.

Example A7: Preparation of Polysaccharide 7 [000160] Didecyl aspartate, para-toluenesulfonic acid salt is obtained according to the process described in US Pat. No. 4,826,818. By a method similar to that described in Example A1, a sodium dextranemethylcarboxylate, synthesized according to the process described in Example A1 using a dextran with a weight average molecular weight of about 10 kg / mol (g). = 38, PHARMACOSMOS), modified with didecyl aspartate is obtained. [000162] According to the dry extract; [Polysaccharide 7] = 19.8 mg / g [000163] Based on acid / base assay: n = 1.05 [000164] From 1 H NMR: m = 0.05.

Example A8: Preparation of Polysaccharide 8 [000165] Dilauryl aspartate, paratoluenesulphonic acid salt is obtained according to the process described in US Pat. No. 4,826,818. By a method similar to that described in Example A1, a sodium dextranemethylcarboxylate, synthesized according to the method described in Example A1 using a dextran with a weight average molecular weight of about 5 kg / mol (g). = 19, PHARMACOSMOS), modified by dilauryl aspartate is obtained. [000167] From the dry extract: [Polysaccharide 8] = 22.8 mg / g [000168] According to the acid / base assay: n = 1.05 [000169] According to the 1H NMR: m = 0.05.

Example A9: Preparation of Polysaccharide 9 [000170] N- (2-aminoethyl) dodecanamide is obtained according to the process described in US Pat. No. 2,387,201 from the methyl ester of dodecanoic acid (SIGMA) and ethylenediamine (ROTH ). By a method similar to that described in Example A1, a sodium dextranemethylcarboxylate, synthesized according to the method described in Example A1 using a dextran with a weight average molecular weight of about 10 kg / mol (g). = 38, PHARMACOSMOS), modified with N- (2-aminoethyl) dodecanamide is obtained. [000172] According to the dry extract: [Polysaccharide 9] = 23.8 mg / g. According to the acid / base assay: n = 1.0 [000174] From 1 H NMR = 0.1.

Example A10: Preparation of Polysaccharide [000175] Sodium dextransuccinate is obtained from a dextran of average molar mass by weight of approximately 10 kg / mol (q = 38, PHARMACOSMOS) according to the method described in the article de Sanchez-Chaves et al., 1998 (Manuel et al., Polymer 1998, 39 (13), 2751-2757). According to 1 H NMR in D 2 0 / NaOD, the average number of succinic groups per glucosidic unit is 1.4. [000176] Lauryl glycinate, a para-toluenesulfonic acid salt, is obtained according to the process described in US Pat. No. 4,826,818. By a method similar to that described in Example A1, a sodium dextransuccinate modified with lauryfe glycinate is obtained. According to the dry extract: [Polysaccharide 10] = 16.1 mg / g [000179] According to the acid / base assay: n = 1.05 [000180] According to the 1 H NMR: m = 0.05.

Example A11: Preparation of Polysaccharide 11 [000181] Dioctyl aspartate, para-toluenesulphonic acid salt is obtained according to the process described in US Pat. No. 4,826,818. 12 g (ie 0.22 mol of hydroxyl) of dextran with a weight average molar mass of approximately 10 kg / mol (q = 38, PHARMACOSMOS) are solubilized in a DMF / DMSO mixture. The mixture is heated to 80 ° C with stirring. 3.32 g (0.03 mol) of 1,4-diazabicyclo [2.2.2] octane then 14.35 g (0.11 mol) of ethyl isocyanatoacetate are progressively introduced. After 5 hours of reaction, the medium is diluted with water and purified by diafiltration on a 5 kD PES membrane against 0.1N NaOH, 0.9% NaCl and water. The final solution is assayed by dry extract to determine the concentration of polysaccharide; then assayed by acid / base assay in 50/50 (V / V) water / acetone to determine the average number of N-methyfcarboxylate carbamate units per glucosidic unit. According to the dry extract [polysaccharide] = 30.5 mg / g [000184] According to the acid / base assay: the average number of N-methylcarboxylate carbamate units per glucoside unit is 1.4. By a method similar to that described in Example A1, a sodium N-methylcarboxylate dextran carbamate modified with dioctyl aspartate is obtained. According to the dry extract [Polysaccharide 11] = 17.8 mg / g [000187] According to the acid / base assay: n = 1.3 [000188] According to the 1H NMR: m = 0.1.

Example Al2: Preparation of Polysaccharide 12 [000189] Dilauryl aspartate, paratoluenesulphonic acid salt is obtained according to the process described in US Pat. No. 4,826,818. By a method similar to that described in Example A1, a sodium dextranemethylcarboxylate, synthesized according to the process described in Ex. Al using a dextran with a weight average molecular weight of about 1 kg / mol (g). = 4, PHARMACOSMOS), modified with dilauryl aspartate is obtained. [000191] According to the dry extract: [Polysaccharide 12] = 12.3 mg / g [000192] According to the acid / base assay: n = 0.95 [000193] According to the 1H NMR: m = 0.07.

Example A13: Preparation of Polysaccharide 13 [000194] 2- (2-Aminoethoxy) ethyl dodecanoate, para-toluenesulphonic acid salt is obtained according to the process described in US Pat. No. 4,826,818. By a method similar to that described in Example A1, a sodium dextranemethylcarboxylate, synthesized according to the process described in Example A1 using a dextran with a weight average molecular weight of about 10 kg / mol (g). = 38, PHARMACOSMOS), modified with 2- (2-aminoethoxy) ethyl dodecanoate is obtained. According to the dry extract: [Polysaccharide 13] = 25.6 mg / g [000197] According to the acid / base assay: n = 1.0 [000198] According to the 1 H NMR m = 0.1.

Example A14: Preparation of Polysaccharide 14 [000199] 2- (2- {2- [dodecanoylamino] ethoxy} ethoxy) ethylamine is obtained according to the process described in US Pat. No. 2,387,201 from methyl ester of dodecanoic acid ( SIGMA) and triethylene glycol diamine (HUNSTMAN). By a method similar to that described in Example A1, a sodium dextranemethylcarboxylate, synthesized according to the method described in Example Al using a dextran with a weight average molecular weight of about 10 kg / mol (g). = 38, PHARMACOSMOS), modified with 2- (2- {2- [2-dodecanoylamino] ethoxy} ethoxy) ethylamine is obtained. According to the dry extract [Polysaccharide 14] = 24.9 mg / g [000202] According to the acid / base assay: n = 1.0 [000203] According to the 1H NMR: m = 0.1.

Example A15: Preparation of Polysaccharide [000204] 2- (2- {2- [hexadecanoylamino] ethoxy} ethoxy) ethylamine is obtained according to the method described in US Pat. No. 2,387,201 from palmitic acid methyl ester (SIGMA ) and triethylene glycol diamine (HUNSTMAN). By a method similar to that described in Example A1, a sodium dextranemethylcarboxylate, synthesized according to the process described in Ex. Al using a dextran with a weight average molecular weight of about 10 kg / mol (g). 38, PHARMACOSMOS), modified with 2- (2- {2- [hexadecanoylamino] ethoxy} ethoxy) ethylamine is obtained. According to the dry extract: [Polysaccharide 15] = 22.2 mg / g [000207] According to the acid / base assay: n = 1.05 [000208] According to the 1H NMR: m = 0.05.

Example A16: Preparation of Polysaccharide 16 [000209] Cetyl glycinate, paratoluenesulfonic acid salt is obtained according to the process described in US Pat. No. 4,826,818. By a method similar to that described in Example A1, a sodium dextranemethylcarboxylate, synthesized according to the process described in Exeple A1 using a dextran with a weight average molecular weight of about 5 kg / mol (g). = 19, PHARMACOSMOS), modified with cetyl glycinate is obtained. According to the dry extract: [Polysaccharide 16] = 23 mg / g [000212] According to the acid / base assay: n = 1.05 [000213] According to the 1H NMR: m = 0 .05.

Example A17 Preparation of Polysaccharide 17 [000214] 10 g (ie 185 mmol of hydroxyls) of dextran with a weight average molar mass of approximately 10 kg / mol (q = 38, PHARMACOSMOS) are solubilized in water at 42 ° C. g / L. To this solution are added 19 ml of 10 N NaOH (185 mmol). The mixture is brought to 35 ° C and then 7.6 g (74 ram) of sodium chloroacetate are added. The temperature of the reaction medium is brought to 60 ° C. at 0.5 ° C./min, then maintained at 60 ° C. for 100 minutes. The reaction medium is diluted with 200 ml of water, neutralized with acetic acid and purified by ultrafiltration on PES membrane 5 kDa against 6 volumes of water. The final solution is assayed by dry extract to determine the concentration of polysaccharide; then assayed by acid / base assay in water / acetone 50/50 (V / V) to determine the average number of methylcarboxylate units per glucosidic unit. According to the dry extract [polysaccharide] = 35.1 mg / g [000216] According to the acid / base assay: the average number of methylcarboxylate units per glucosidic unit is 0.42. The sodium dextranmethylcarboxylate solution is passed through a Purolite resin (anionic) to obtain the acid dextran-methylcarboxylic acid which is then lyophilized for 18 hours. [000218] Cetyl glycinate, para-toluenesulfonic acid salt is obtained according to the process described in US Pat. No. 4,826,818. By a method similar to that described in Example A1, a cetyl glycinate modified sodium dextranemethylcarboxylate is obtained. [000220] According to the dry extract: [Polysaccharide 17] = 18 mg / g [000221] According to the acid / base assay: n = 0.37 [000222] According to the 1H NMR: m = 0.05.

[000223] Examples Part B Demonstration of the Properties of the Compositions According to the Invention Example BI: Rapid Analogous Insulin Solution (Novolog®) at 100 IU / mL [000224] This solution is a commercially available commercial insulin aspart solution by Novo Nordisk under the name of Novolog® in the USA and Novorapid® in Europe. This product is a fast analog insulin.

Example B2: Fast Analogous Insulin Solution (Humalog) at 100 IU / mL [000225] This solution is a commercial solution of Lispro insulin marketed by Eli Lilly under the name Humalog®. This product is a fast analog insulin.

Example B3: Rapid Analogue Insulin Solution (Apidra) at 100 IU / mL [000226] This solution is a commercial solution of insulin glulisine marketed by Sanofi-Aventis under the name Apidra® This product is a rapid analog insulin .

Example B4: Slow Analogous Insulin Solution (Lantus) at 100 IU / mL [000227] This solution is a commercial solution of insulin Glargine marketed by Sanofi-Aventis under the name of Lantus. This product is a slow analog insulin.

Example B5: Human Insulin Solution (ActRapid) at 100 IU / mL [000228] This solution is a commercial solution of Novo Nordisk sold under the name Actrapid®. This product is a human insulin. Example B6 Solubilization of Lantus at 100 IU / mL and pH 7 with dextran substituted [000229] 20 mg of polysaccharide 4 described in Example A4 are accurately weighed. This lyophilizate is taken up in 2 ml of Lantus® in its commercial formulation. A temporary precipitate appears but the solution becomes clear after about 30 minutes. The pH of this solution is 6.3. The pH is adjusted to 7 with a 0.1N sodium hydroxide solution. This clear solution is filtered over 0.22 μm and is then placed at + 4 ° C.

Example B7: Preparation of a Substituted Dextran / Lantus® / Apidra® 75/25 Composition at pH 7 [000230] To 0.75 ml of the polysaccharide 4 / Lantus® solution prepared in Example B6 is added 0, 25 ml of Apidra® (in its commercial formulation) to form 1 ml of a composition at pH 7. The composition is clear attesting to the good solubility of Lantus® and Apidra® under these formulation conditions. This clear solution is filtered through 0.22 μm and then placed at + 4 ° C.

Example B8: Preparation of a substituted dextran / Lantus® / Humalog® 75/25 composition at pH 7 [000231] To 0.75 mL of the polysaccharide 4 / Lantus® solution prepared in Example B6 is added 0, 25 ml of Humalog® (in its commercial formulation) to form 1 ml of a composition at pH 7. The composition is clear attesting to the good solubility of Lantus® and Humalog® under these formulation conditions. This clear solution is filtered over 0.22 μm and is then placed at + 4 ° C.

Example B9: Preparation of a substituted dextran / Lantus® / Novolog® 75/25 composition at pH 7 [000232] To 0.75 mL of the polysaccharide 4 / Lantus® solution prepared in Example B6 is added 0.25 mL of Novolog® (in its commercial formulation) to form 1 mL of a composition at pH 7. The composition is clear attesting the good solubility of Lantus® and Novolog® under these conditions of formulation. This clear solution is filtered over 0.22 μm and is then placed at + 4 ° C.

Example B10 Preparation of a Substituted Dextran / Lantus® / ActRapid® 75/25 Composition at pH 7 [000233] To 0.75 mL of the polysaccharide 4 / Lantus® solution prepared in Example B6 was added 0.25 mL ActRapid® (in its commercial formulation) to form 1 mL of a composition at pH 7. The composition is clear attesting to the good solubility of Lantus® and ActRapid® under these formulation conditions. This clear solution is filtered over 0.22 μm and is then placed at + 4 ° C.

Example B11 Preparation of a Substituted Dextran / Lantus® / Apidra® 60/40 Composition at pH 7 [000234] To 0.6 ml of the polysaccharide 4 / Lantus® solution prepared in Example B6 is added 0.4 ml of Apidra® (in its commercial formulation) to form 1 ml of a composition at pH 7. The composition is clear attesting to the good solubility of Lantus® and Apidra® under these formulation conditions. This clear solution is filtered over 0.22 μm and is then placed at + 4 ° C.

Example B12: Preparation of a Substituted Dextran / Lantus / Apidra® 40/60 Composition at pH 7 [000235] To 0.4 ml of the polysaccharide 4 / Lantus ° solution prepared in Example B6 is added 0.6 ml of Apidra ° (in its commercial formulation) to form 1 ml of a composition at pH 7. The composition is clear attesting to the good solubility of Lantus® and Apidra ° under these conditions of formulation. This clear solution is filtered over 0.22 μm and is then placed at + 4 ° C.

Example B13: Precipitation of Lantus® [000236] 1 ml of Lantus® is added in 2 ml of a solution of PBS containing 20 mg / ml of BSA (bovine serum albumin). The PBS / BSA mixture simulates the composition of the subcutaneous medium. A precipitate appears which is in good agreement with Lantus' mechanism of operation (injection precipitation due to increased pH). [000237] Centrifugation at 4000 rpm is carried out to separate the precipitate from the supernatant. Then Lantus® is assayed in the supernatant. As a result, 86% of Lantus ° is found in a precipitated form.

Example B14: Precipitation of a Substituted Dextran / Lantus® Composition [000238] 1 ml of polysaccharide 4 / Lantus solution prepared in Example B6 is added in 2 ml of a PBS solution containing 20 mg / ml of BSA (bovine) albumin serum). The PBS / BSA mixture simulates the composition of the subcutaneous medium. A precipitate appears. Centrifugation at 4000 rpm is performed to separate the precipitate from the supernatant. Then Lantus® is assayed in the supernatant. As a result, 85% of Lantus ° is found in a precipitated form. This precipitation percentage of Lantus® is identical to that obtained for the control described in Example B13.

Solubilization and precipitation tests identical to those described in Examples B6 and B14 were carried out with other dextrans substituted at the same concentration of 10 mg / ml of polysaccharide per 100 IU / ml of Lantus®. 20 mg of polysaccharide in lyophilizate form are precisely weighed. This lyophilizate is taken up in 2 ml of Lantus® in its commercial formulation. A temporary precipitate appears but the solution becomes clear after about 30 minutes to a few hours (depending on the nature of the polysaccharide). The pH of this solution is 6.3. The pH is adjusted to 7 with 0.1 N sodium hydroxide solution. This clear solution is filtered through 0.22 μm and then placed at + 4 ° C. The results are summarized in Table 2.

Polysaccharide No. Solubilization Lantus® Precipitation% of Lantus® Precipitation 2 Yes Yes 85 1 Yes Yes No Measured 4 Yes Yes 87 3 Yes Yes No Measured Yes Yes 94 6 Yes Yes No Measured 7 Yes Yes No Measured 8 Yes Yes No Measured 9 Yes Yes 94 Yes Yes No measured Yes Yes No measured 14 Yes Yes No measured 13 Yes Yes No measured 12 Yes Yes No measured 11 Yes Yes No measured 16 Yes Yes No measured 17 Yes Yes No measured Table 2 5 Example B15: Precipitation d substituted dextran / Lantus® / Apidra® 75/25 composition at pH 7 1 mL of the composition Dextran substituted / Lantus® / Apidra® 75/25 (containing 7.5 mg / mL polysaccharide 75 IU / mL Lantus® and 25 IU / ml of Apidra®) prepared in Example B7 is added in 2 ml of a solution of PBS containing 20 mg / ml of BSA (bovine serum albumin). The PBS / BSA mixture simulates the composition of the subcutaneous medium. A precipitate appears. [000241] Centrifugation at 4000 rpm is carried out to separate the precipitate from the supernatant. Then Lantus® is assayed in the supernatant. The precipitation percentages of Lantus® are similar to the control described in Example B13. Example B16: Precipitation of Different Compositions by Varying the Nature of the Polysaccharide Other tests under the same conditions as those of Example B15 were carried out in the presence of other polysaccharides. The results are grouped in the following Table 3 and it is observed that the solubilization and the precipitation of Lantus® are preserved. Polysaccharide no. Solubilization Percentage of Lantus® / Apidra® precipitation 75/25 2 Yes 85 1 Yes No measured 4 Yes 87 3 Yes No measured 5 Yes 86 6 Yes No measured 7 Yes No measured 8 Yes No measured 9 Yes 86 Yes 85 Yes 87 14 Yes 86 13 Yes 88 12 Yes 91 10 Table 3

Example B17: Precipitation of Different Compositions by Varying the Nature of Meally Insulin [000244] Compositions are prepared by mixing 0.75 mL of the Polysaccharide 4 / Lantus® solution prepared in Example B6 with 0.25. mL of a mealtime insulin to form 1 mL of substituted dextran / Lantus® / mealtime insulin (containing 7.5 mg / mL polysaccharide, 75 IU / mL Lantus® and 25 IU / mL mealtime insulin) [000245 This composition is added in 2 ml of PBS containing 20 mg / ml of BSA (bovine serum albumin). The PBS / BSA mixture simulates the composition of the subcutaneous medium. A precipitate appears. Centrifugation at 4000 rpm is performed to separate the precipitate from the supernatant. Then Lantus® is assayed in the supernatant. In the presence of the 4 prandial insulins tested, Lantus® precipitates at about 90%. This percentage of precipitation of Lantus® is similar to the control described in Example B13, the results are summarized in Table 4. Nature of Prandial Insulin Solubilization Percentage of Lantus® Precipitation / Lantus® Prandial Insulin 75/25 Apidra® Yes 88 Novolog® Yes 92 Humalog® Yes 89 ActRapid® Yes 90 Table 4 Example B18: Chemical stability of the compositions [000247] The composition Dextran substituted / Lantus® / Apidra® 75/25 at pH 7 described in Example B7 as well as the corresponding controls are placed at 30 ° C for 4 weeks. The regulation requires 95% native (undegraded) insulin after 4 weeks at 30 ° C. After 4 weeks, the formulation meets the specifications defined by the regulations, the results are summarized in Table 5. Compositions Percentage of native Lantus® Percentage of Apidra® after 4 weeks at 30 ° C native after 4 weeks at 30 ° C. Lantus® (commercial formulation 97 na) Apidra® (commercial formulation n. 95) composition Dextran 96 substituted 98 / Lantus® / Apidra® 75/25 at pH 7 Table 5 [000249] A percentage of insulin is thus obtained native of 96.5%, which complies with the requirements of the regulations.

Example B19: Xnjectability of the Solutions [000250] All the compositions prepared are injectable with the usual insulin injection systems. The solutions described in Examples B7 to B13 and the compositions described in Examples B15 to B17 are injected without any difficulty with insulin syringes with 31 Gauge needles with Novo Nordisk insulin pens marketed under the named Novopen®, equipped with 31 Gauge needles. Example B20: Protocol for Measuring the Pharmacodynamics of Insulin Solutions Pre-clinical studies were carried out on pigs in order to evaluate two compositions according to the invention: Lantus® / Apidra® (75/25), formulated with Polysaccharide 4 (6 mg / mL), and [000252] Lantus® / Humalog® (75/25), formulated with Polysaccharide 4 (6 mg / mL).

[000253] The hypoglycemic effects of these compositions were compared with injections made with simultaneous but separate injections of Lantus® (pH 4) and then of a prandial insulin Apidra® or Humalog®.

[000254] Six domestic pigs of about 50 kg, previously catheterized at the jugular, are fasted 2 to 3 hours before the start of the experiment. In the hour before the injection of insulin, 3 blood samples are taken to determine the basal glucose level. [000255] Insulin injection at a dose of 0.4 IU / kg is performed by subcutaneous injection in the neck, under the animal's ear using the Novopen® insulin pen equipped with of a G 31 needle. [000256] Blood samples are then taken after 4, 8, 12, 16, 20, 30, 40, 50, 60, 90, 120, 240, 360, 480, 600, 660 and 720 minutes. After each sample, the catheter is rinsed with a dilute solution of heparin. [000257] A drop of blood is taken to determine the blood glucose by means of a glucometer. The glucose pharmacodynamics curves are then plotted. The results obtained are presented in the form of the glucose pharmacodynamics curves shown in FIGS. 1-6. [000259] Lantus® / Apidra® (75/25), formulated with Polysaccharide 4 (6 mg / ml). [000260] FIG. 1: Average curves + standard deviation of the average for the sequential administrations of Apidra® and Lantus® in comparison with a composition according to the invention Polysaccharide 4 / Lantus® / Apidra® (75/25) [000261] Figure 2: Individual Apidra® Lantus® Curves [000262] Figure 3: Individual Polysaccharide 4 / Apidra® / Lantus® Curves [000263] Figure 1 shows the mean blood glucose drop curves as well as the standard deviations of the mean for blood glucose. pigs tested for each formulation. The drop in blood glucose in the first 30 minutes is similar for both formulations indicating that the presence of a polysaccharide does not interfere with the rapidity of Apidra®. On the other hand, between 90 min and 10 h (600 minutes) the sequential administration of Apidra® Lantus® induces a heterogeneous glucose drop with a homogeneous plateau response on three pigs and a heterogeneous response on the other three pigs ( Figure 2). On the contrary, the 6 pigs tested with the formulation Polysaccharide 4 / Apidra® / Lantus® have a homogeneous response (FIG. 3). This results in the analysis of the coefficients of variation (CV) between 60 min and 10 h which are on average 54% (between 21% and 113%) for the Apidra Lantus ° control and 12% (between 5% and 25%) for Polysaccharide 4 / Apidra® / Lantus®. [000265] Lantus® / Humalog® (75/25), formulated with Polysaccharide 4 (6 mg / ml). [000266] FIG. 4: Average Curves + Standard Deviation for the Sequential Administrations of Humalog and Lantus Compared with the Administration of a Composition According to the Invention Polysaccharide 4 / Humalog® / Lantus® [000267] FIG. Figure 6: Individual curves Polysaccharide 4 / Humalog® / Lantus® [000269] Figure 4 shows the mean curves of blood glucose drop as well as the standard deviations of the average for the pigs tested. on each formulation. The drop in blood glucose in the first 30 minutes is similar for both formulations indicating that the presence of Polysaccharide 4 does not interfere with the rapidity of Humalog®. In contrast, between 60 min and 8 h (480 min) Sequential administration of Humalog® and Lantus® induced a heterogeneous glucose drop with a homogeneous plateau response on four pigs and a heterogeneous response on the other two pigs (Figure 5). ). In contrast, the pigs tested with the Polysaccharide 4 / Humalog® / Lantus® formulation had a homogeneous response (Figure 6). This results in the analysis of the coefficients of variation (CV) on blood glucose drop data between 60 min and 8h, which averages 54% (between 31% and 72%) for the Humalog® Lantus® control and 15% (between 6% and 28%) for Polysaccharide 4 / Humalog® / Lantus®, The presence of polysaccharide 4 strongly reduces the variability of Lantus® on the drop in blood glucose. 15 20 25 30

Claims (22)

REVENDICATIONS1. A composition in the form of an injectable aqueous solution, the pH of which is between 6.0 and 8.0, comprising at least: a) a basal insulin whose isoelectric point pI is between 5.8 and 8.5; b) a dextran substituted with radicals carrying carboxylate charges and hydrophobic radicals of formula I: - Formula I in which - R is -OH or chosen from the group consisting of the radicals o - (f- [A] -COOH) o - (g- [B] -k- [D]), D having at least one alkyl chain having at least 8 carbon atoms, n represents the degree of substitution of glucosidic units by -F- [A] -COOH and 0.1 n 2; m represents the degree of substitution of the glucoside units by -G- [B] -k- [D] and 0 <m 0.5; q represents the degree of polymerization in glucosidic units, that is to say the average number of glucoside units per polysaccharide chain and g5. - - (f- [A] -COOH) r, o -A- is a linear or branched radical having 1 to 4 carbon atoms; said radical -A-: o being bound to a glucosidic unit by a function f selected from the group consisting of ether, ester and carbamate functions. - (g- [B] -k- []]) m, o -B- is an at least divalent linear or branched radical comprising 1 to 4 carbon atoms; said radical -B-: o being bound to a glucosidic unit by a function g selected from the group consisting of ether, ester and carbamate functions; o being bound to a radical -D by a function k; k selected from the group consisting of ester, amide, and carbamate; said radical - D: - being a radical -X (-IY) p, X being an at least divalent radical comprising from 1 to 12 atoms selected from the group consisting of C, N or O atoms, optionally carrying carboxyl functions or amine and / or derived from an amino acid, a dihydric alcohol, a diamine or a mono- or polyethylene glycol mono- or diamine; Y being a linear or cyclic alkyl group, a C8 to C20 alkylaryl or arylalkyl, optionally substituted by one or more C1 to C3 alkyl groups; p? 1 and / a function selected from the group consisting of ester, amide and carbamate functions. f, g and k being identical or different. the free acid functions being in the form of salts of alkaline cations selected from the group consisting of Na 'and K. - and when p = 1, if Y is C8-C14 alkyl, then q * m? 2, if Y is C15 alkyl, then q * m> 2; and if Y is C16-C20 alkyl, then q * m? 1 - and when p? 2, if Y is C8 to C11 alkyl, then q * m? 2 and, if Y is C12-C16 alkyl, then q * m? 0.3. 2. Composition according to claim 1, characterized in that the radical - (f- [A] -COOH) is chosen from the group consisting of the following sequences, (having the meaning given above). 3. Composition according to any one of the preceding claims, characterized in that the radical - (g- [B] -k- [D]) n, is chosen from the group consisting of the following sequences; g, k and D having the meanings given above k, 25 4. Composition according to any one of the preceding claims, characterized in that the radical - (f- [A] -COOH) 'is such that - -A- is a radical comprising 1 carbon atom; said radical -A- being attached to a glucosidic unit by a function of ether. 5. Composition according to any one of the preceding claims, characterized in that the radical - (g- [B] -k- [D]) m is such that - -B- is a radical comprising 1 carbon atom; said radical -B- being attached to a glucoside unit by a g ether function, and, - X is a radical derived from an amino acid. 6. Composition according to any one of the preceding claims, characterized in that the radical - (f- [A] -COOH) 'is such that - is a radical comprising 1 carbon atom; said radical -A- being linked to a glucoside unit by a function f ether, and, the radical - (g- [B] -k- [D]), is such that: -B- is a radical comprising 1 atom of carbon ; said radical -B- being linked to a glucoside unit by a g ether function, and, - X is a radical derived from an amino acid. k is an amide function. 7. Composition according to any one of the preceding claims, characterized in that the radical X is an at least divalent radical derived from an amino acid selected from the group consisting of glycine, phenylalanine, lysine and acid. aspartic. 8. Composition according to any one of claims 1 to 4, characterized in that the radical X is at least one radical cleaved from ethylene glycol. 9. Composition according to any one of claims 1 to 4, characterized in that the radical X is an at least divalent radical derived from ethylenediamine. 10. Composition according to any one of claims 1 to 4, characterized in that the radical X is an at least divalent radical derived from a polyethylene glycol selected from the group consisting of diethylene glycol and triethylene glycol. 11. Composition according to any one of claims 1 to 4, characterized in that the radical X is an at least divalent radical derived from a mono- or polyethylene glycol amine selected from the group consisting of ethanolamine, diethylene glycol amine and triethylene glycol amine. 12. Composition according to any one of claims 1 to 4, characterized in that the radical X is an at least divalent radical derived from a mono- or polyethylene glycol diamine selected from the group consisting of diethylene glycol diamine and triethylene glycol diamine. 13. Composition according to any one of the preceding claims, characterized in that the group Y is an alkyl group derived from a hydrophobic alcohol, selected from the group consisting of octanol (caprylic alcohol), 3,7-dimethyloctan -laugh out loud ; decanol (decyl alcohol), dodecanol (lauryl alcohol), tetradecanol (meryl alcohol) and hexadecanol (cetyl alcohol). 14. Composition according to any one of claims 1 to 12, characterized in that the group Y is an alkyl group derived from a hydrophobic acid, selected from the group consisting of decanoic acid, dodecanoic acid, tetradecanoic acid and hexadecanoic acid. 15. Composition according to any one of the preceding claims, characterized in that the basal insulin whose isoelectric point is between 5.8 and 8.5 is insulin glargine. 16. Composition according to any one of the preceding claims, characterized in that it further comprises a mealtime insulin. 17. Composition according to any one of the preceding claims, characterized in that it further comprises zinc salts at a concentration of between 0 and 1000 IJM. 18. Composition according to any one of the preceding claims, characterized in that it comprises buffers chosen from the group comprising Tris and phosphates at concentrations of between 0 and 100 mM, preferably between 15 and 50 mM. 19. Composition according to any one of claims 16 to 18, characterized in that said prandial insulin is selected from the group consisting of human insulin, insulin glulisine, insulin lispro and insulin aspart. 20. A one-dose formulation containing a composition according to any one of claims 1 to 15, at a pH of between 7 and 7.8 and a mealtime insulin. 21. Single-dose formulation according to claim 20, characterized in that the prandial insulin is selected from the group comprising human insulin. 22. Single-dose formulation according to claim 20, characterized in that the prandial insulin is selected from the group comprising insulin lispro, insulin glulisine and insulin aspart.