DESC:PRIORITY DOCUMENTS
This application claims priority from the Indian provisional application number IN 201621032421; filed on Sep 22, 2016.

TECHNICAL FIELD OF THE INVENTION
The invention relates to an aqueous pharmaceutical composition comprising: (a) from about 1 IU/ml to about 500 IU/ml of buffered insulin glargine, (b) therapeutically effective amount of a GLP-1 analogue, and (c) one or more pharmaceutically acceptable excipients. In particular, the invention relates to an aqueous pharmaceutical composition comprising: (a) liraglutide and buffered insulin glargine in a weight ratio of about 1:1 to about 1:20, and (b) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 6 to about 9.

BACKGROUND OF THE INVENTION
Diabetes mellitus is a metabolic disorder in which the ability to utilize glucose is partly or completely lost. Since the introduction of insulin in the 1920's, continuous efforts have been made to improve the treatment of diabetes mellitus. Since people suffering from diabetes are subject to chronic treatment over several decades, there is a major need for safe, convenient and life quality improving insulin formulations.
In the treatment of diabetes mellitus, many varieties of insulin formulations have been suggested and used, such as regular insulin, isophane insulin (designated NPH), insulin zinc suspensions (such as SEMILENTE®, LENTE®, and ULTRALENT®), and biphasic isophane insulin. Some of the commercial available insulin formulations are characterized by a fast onset of action and other formulations have a relatively slow onset but show a more or less prolonged action. Fast-acting insulin formulations are usually solutions of insulin, while retarded acting insulin formulations can be suspensions containing insulin in crystalline and/or amorphous form precipitated by addition of zinc salts alone or by addition of protamine or by a combination of both. Within the two decade a number of human insulin analogues have been developed. They are designed for particular profiles of action, i.e. fast acting or prolonged action.
Another peptide which has very important in the treatment of diabetes is glucagon-like peptide-1 (GLP-1). Human GLP-1 is a 37 amino acid residue peptide originating from preproglucagon which is synthesized in the L-cells in the distal ileum, in the pancreas and in the brain. GLP-1 is an important gut hormone with regulatory function in glucose metabolism and gastrointestinal secretion and metabolism. GLP-1 stimulates insulin secretion in a glucose-dependant manner, stimulates insulin biosynthesis, and promotes beta cell rescue, decreases glucagon secretion, gastric emptying and food intake. As the type 2 diabetes population is rapidly increasing in the world there is a much larger need for simpler administration of more effective drugs.

US Patent Application No. 20120021978 A1 discloses a composition of GLP-1 agonist and Insulin peptide. US Patent Application No. 20120295846 A1 discloses a composition of GLP-1 agonist and Insulin with Methionine. US Patent Application No. 20120122774 A1 discloses composition comprising insulin glargine and analogue of exedin-4. PCT publication WO 2006/051103 A2 discloses pharmaceutical composition comprising insulinotropic peptide and basal insulin. PCT Publication No. WO 1995031214 A1 discloses a treatment of insulin requiring diabetes comprising administration of insulin and GLP-1. PCT Publication No. WO 2003020201 A3 discloses pre-mixed formulation of GLP-1 compounds and basal insulin. PCT Publication No. WO 2017145104 A1 discloses a pharmaceutical composition of insulin glargine with two amino acids and another insulin analogue.
There is a need to develop a pharmaceutical composition of insulin glargine with liraglutide which can provide alternate treatment option to diabetic patients.

SUMMARY OF THE INVENTION
In one aspect of the invention, there is provided an aqueous pharmaceutical composition comprising: (a) from about 1 IU/ml to about 500 IU/ml of buffered insulin glargine, (b) therapeutically effective amount of a GLP-1 analogue, and (c) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 3 to about 9. In an embodiment, the GLP-1 analogue comprises exenatide, liraglutide, lixisenatide, semaglutide, dulaglutide and albiglutide. Preferably, the GLP-1 analogue is liraglutide, present in the amount from about 0.3 mg/ml to about 2 mg/ml. In another embodiment, the buffered insulin glargine is present in about 40IU/mL to about 500IU/mL. In another embodiment, the one or more pharmaceutically acceptable excipients comprise a pH maintaining agent, solubilising agent, isotonic agent, preservative, pH modifying agent or combination thereof. In another embodiment, the composition is stable for at least 24 hours when stored at 25 °C and relative humidity of 60%. In another embodiment, the composition is stable for at least 6 months when stored at 25 °C and relative humidity of 60%. In still another embodiment, the buffered insulin glargine comprises insulin glargine and two amino acids viz., arginine and isoleucine. In yet another embodiment, the two amino acids, arginine and isoleucine are present in a weight ratio of about 1:1 to about 1:5, preferably about 1:2.

The pharmaceutical composition of the present invention is suitable for parenteral administration such as intramuscular, subcutaneous, intradermal and intravenous administration. Preferably, the pharmaceutical composition is suitable for subcutaneous injection administration.

In another aspect of the invention, there is provided an aqueous pharmaceutical composition comprising: (a) from about 100 IU/ml of buffered insulin glargine, (b) therapeutically effective amount of liraglutide, (c) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 6 to about 9. In one embodiment, the buffered insulin glargine and liraglutide are present separated from each other or as a mixture. In another embodiment, the buffered insulin glargine and liraglutide are formulated as a fixed dose composition.

In one embodiment of first aspect, there is provided an aqueous pharmaceutical composition comprising: (a) about 100 IU/ml of buffered insulin glargine, (b) about 0.6 mg/mL of liraglutide, and (c) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 3 to about 9.

In another aspect of the invention, there is provided an aqueous pharmaceutical composition comprising: (a) liraglutide and buffered insulin glargine in a weight ratio of about 1:1 to about 1:20, and (b) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 6 to about 9.

In one embodiment, the one or more pharmaceutically acceptable excipients comprise a pH maintaining agent, solubilising agent, isotonic agent, preservative, pH modifying agent or combination thereof.
In yet another embodiment, there is provided an aqueous pharmaceutical composition comprising: (a) about 100 IU/ml of buffered insulin glargine, (b) about 0.3 mg/mL to about 2 mg/mL of liraglutide, and (c) one or more pharmaceutically acceptable excipients selected from a pH maintaining agent, solubilising agent, isotonic agent, preservative, pH modifying agent or combination thereof, wherein the pharmaceutical composition has a pH between about 3 to about 9. In another embodiment, amount of liraglutide is about 0.6 mg/mL or about 1.2 mg/mL or about 1.8 mg/mL.

In yet another embodiment, there is provided an aqueous pharmaceutical composition comprising: (a) 3.64 mg of buffered insulin glargine, (b) about 0.3 mg of liraglutide, (c) 10 mg of 85% glycerol, (d) 30 µg of zinc Chloride, (e) 2.7 mg of m-cresol, (f) 5mM of L-arginine and 10 mM of isoleucine and (g) 1 mg of dipotassium phosphate, and wherein the pharmaceutical composition has a pH of 8.2 ± 0.2.

In yet another embodiment, there is provided an aqueous pharmaceutical composition comprising: (a) 3.64 mg of buffered insulin glargine, (b) about 0.6 mg of liraglutide, (c) 10 mg of 85% glycerol, (d) 30 µg of zinc Chloride, (e) 2.7 mg of m-cresol, (f) 5mM of L-arginine and 10 mM of isoleucine and (g) 1 mg of dipotassium phosphate, and wherein the pharmaceutical composition has a pH of 8.2 ± 0.2.

In another aspect of this invention, there is provided an aqueous pharmaceutical composition comprising first component and second component, wherein said first component comprises (a) buffered insulin glargine, and (b) a pharmaceutically acceptable excipient, and said second component comprises (a) liraglutide, and (b) a pharmaceutically acceptable excipient. In one embodiment, the first component and second component can be administered as mixture or separately. In another embodiment, the composition is formulated as a fixed dose composition. In another embodiment, pharmaceutically acceptable excipients from first and second component are selected from a pH maintaining agent, solubilising agent, isotonic agent, preservative, pH modifying agent or combination thereof

In another aspect of the invention, there is provided a kit comprising an aqueous composition comprising first component and second component, wherein said first component comprises (a) buffered insulin glargine, and (b) a pharmaceutically acceptable excipient, and said second component comprises (a) liraglutide, and (b) a pharmaceutically acceptable excipient. In another embodiment, the composition is formulated as a fixed dose composition. In another embodiment, pharmaceutically acceptable excipients from first and second component are selected from a pH maintaining agent, solubilising agent, isotonic agent, preservative, pH modifying agent or combination thereof.

In another aspect of this invention, there is provided a method of treating diabetes mellitus in a subject, said method comprises parenterally administering to said subject an aqueous pharmaceutical composition comprising: (a) from about 1 IU/ml to about 500 IU/ml of buffered insulin glargine, (b) therapeutically effective amount of a GLP-1 analogue, and (c) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 3 to about 9. In an embodiment, said method comprises parenterally administering to said subject an aqueous pharmaceutical composition comprising: (a) liraglutide and buffered insulin glargine in a weight ratio of about 1:1 to about 1:20, and (b) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 6 to about 9.

In another aspect of this invention, there is provided a use of the aqueous composition of any aspect of this invention for the preparation of medicament for treating diabetes, for adjusting the fasting, postprandial and/or post absorptive blood glucose concentration, for improving glucose tolerance, for preventing hypoglycemia, for preventing loss of function of the pancreatic ß-cells and for weight loss and/or for preventing weight gain in a patient in need thereof.

In another aspect of this invention, there is provided a process of using the kit comprising aqueous composition comprising first component and second component, wherein said first component comprises (a) buffered insulin glargine, and (b) a pharmaceutically acceptable excipient, and said second component comprises (a) liraglutide, and (b) a pharmaceutically acceptable excipient, wherein the process comprises the steps of:
(i) selecting a dose of the buffered insulin glargine that is to be administered,
(ii) selecting a dose of the liraglutide that is to be administered,
(iii) determining and administering an amount which corresponds to the doses of buffered insulin glargine and liraglutide.

In another aspect of this invention, there is provided a method of preparing a pharmaceutical composition in the form of a solution, said method comprising the steps of:
(i) preparing a solution comprising insulin glargine in acidic water for injection,
(ii) preparing a solution of at least two amino acids,
(iii) adding the solution of at least two amino acids in the solution of step (i) and adjusting the pH to about 3 to about 4 using a pH modifying agent,
(iv) preparing a solution comprising liraglutide, and
(v) mixing the solutions of step (iii) and (iv) and adjusting the pH to about 8.2± 0.2 to obtain the pharmaceutical composition.

In another embodiment, there is provided a method of preparing a pharmaceutical composition in the form of a suspension, said method comprising the steps of:
(i) preparing a solution comprising insulin glargine in acidic water for injection,
(ii) preparing a solution of at least two amino acids,
(iii) adding the solution of at least two amino acids in the solution of step (i) and adjusting the pH to about 6 to about 9 using a pH modifying agent,
(iv) preparing a solution comprising liraglutide, and
(v) mixing the solutions of step (iii) and (iv) and adjusting the pH to about 8.2± 0.2 to obtain the pharmaceutical composition.

DETAILED DESCRIPTION OF THE INVENTION

The general aspect of the invention relates to an aqueous pharmaceutical composition comprising: (a) buffered insulin glargine, (b) a GLP-1 analogue, and (c) a pharmaceutically acceptable excipient.

The term “aqueous pharmaceutical composition” herein relates to a liquid formulation suitable for parenteral administration such as intramuscular, subcutaneous, intradermal and intravenous administration.

As used herein the term “buffered insulin glargine” refers to insulin glargine and two amino acids in weight ratio of about 1:1 to about 1:5, wherein two amino acids selected from the group consisting of arginine, lysine, betaine, norleucine, aspartic acid, glutamic acid, asparagines, proline, glutamine, histidine, serine, threonine, glycine, alanine, methionine, leucine, isoleucine, valine, phenylalanine, tryptophan, phosphatidylethanolamine, phosphatidylserine and tyrosine. Preferably, the buffered insulin glargine contains insulin glargine and two amino acids viz., arginine and isoleucine in a weight ratio of about 1:2.

The amount of 100 IU/mL of insulin glargine corresponds to 3.64 mg of insulin glargine.

As used herein the term “GLP-1 analogue” refers to glucagon-like peptide-1 receptor agonists also known as GLP-1 receptor agonists or incretin mimetics. GLP-1 analogue comprises exenatide, liraglutide, lixisenatide, semaglutide, dulaglutide and albiglutide.

The term “pharmaceutically acceptable excipients” herein relates to non-active pharmaceutical ingredients which are within the scope of sound medical judgment suitable for use in pharmaceutical products.

The term “stable” herein relates to a physical and/or chemical stability of pharmaceutical composition of insulin and/or Liraglutide, when stored at certain conditions of temperature and relative humidity.

In one aspect of the invention, there is provided an aqueous pharmaceutical composition comprising: (a) from about 1 IU/ml to about 500 IU/ml of buffered insulin glargine, (b) therapeutically effective amount of a GLP-1 analogue, and (c) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 3 to about 9. In an embodiment, the GLP-1 analogue comprises exenatide, liraglutide, lixisenatide, semaglutide, dulaglutide and albiglutide. Preferably, the GLP-1 analogue is liraglutide, present in the amount from about 0.3 mg/ml to about 2 mg/ml. In another embodiment, the buffered insulin glargine is present in about 40IU/mL to about 500IU/mL. In another embodiment, the one or more pharmaceutically acceptable excipients comprise a pH maintaining agent, solubilising agent, isotonic agent, preservative, pH modifying agent or combination thereof. In another embodiment, the composition is stable for at least 24 hours when stored at 25 °C and relative humidity of 60%. In another embodiment, the composition is stable for at least 6 months when stored at 25 °C and relative humidity of 60%. In still another embodiment, the buffered insulin glargine comprises insulin glargine and two amino acids viz., arginine and isoleucine. In yet another embodiment, the two amino acids, arginine and isoleucine are present in a weight ratio of about 1:1 to about 1:5, preferably about 1:2.

In another embodiment, buffered insulin glargine is present in about 40IU/mL to about 500IU/mL. Alternatively, buffered insulin glargine is present in about 80IU/mL to about 400IU/mL, or about 100IU/mL to about 300IU/mL, or about 200IU/mL to about 300IU/mL. Alternatively, buffered insulin glargine is present in about 50IU/mL, or about 60IU/mL, or about 70IU/mL, or about 80IU/mL, or about 90IU/mL, or about 100IU/mL, or about 110IU/mL, or about 120IU/mL, or about 130IU/mL, or about 140IU/mL, or about 150IU/mL, or about 160IU/mL, or about 170IU/mL, or about 180IU/mL, or about 190IU/mL, or about 200IU/mL, or about 250IU/mL, or about 300IU/mL, or about 350IU/mL, or about 400IU/mL, or about 450IU/mL. Each of this concentration constitutes an alternate embodiment of the invention.

In another embodiment, the GLP-1 analogue is liraglutide, present in the amount from about 0.3 mg/ml to about 2 mg/ml or about 06 mg/mL to about 1.8 mg/mL or about 1.2 mg/mL. Each of these amounts constitutes an alternate embodiment of the invention.

In one another embodiment, the pharmaceutical composition has a pH in between about 3 to about 9. Alternatively, the composition has pH in between about 4 to about 8, or about 5 to about 7, or about 6 to about 7, or about 7 to about 8, or about 8 to about 9. Each of the pH range constitutes an alternate embodiment of the invention.

In another embodiment, the one or more pharmaceutically acceptable excipients comprise pH maintaining agent, solubilising agent, isotonic agent, preservative, pH modifying agent or combination thereof.

The pH maintaining agent as used herein include, but are not limited to, phosphate, acetate, citrate or TRIS (i.e. 2-amino-2-hydroxymethyl-1,3-propanediol) and corresponding salts.

The solubilizing agent are selected from the group consisting of partial and fatty acid esters and ethers of polyhydric alcohols such as of glycerol, sorbitol, glycine and the like (Span®, Tween®, in particular Tween® 20 and Tween®80, Myrj®, Brij®, Cremophore® or poloxamers, Pluronics® and Tetronics®), polysorbates (Tween™), sodium dodecyl sulfate (sodium lauryl sulfate), lauryl dimethyl amine oxide, cetyltrimethylammonium bromide (CTAB), polyethoxylated alcohols polyoxyethylene sorbitan, Octoxynol (Triton X100™), N, N - dimethyldodecylamine-N-oxide, hexadecyltrimethylammonium bromide (HTAB), polyoxyl 10 lauryl ether, Brij 721™, bile salts (sodium deoxycholate, sodium cholate), polyoxyl castor oil (Cremophor™), nonylphenol ethoxylate (Tergitol™), cyclodextrins, lecithin and methylbenzethonium chloride (Hyamine™). The concentration of solubilizing agent is about 1mM to 200mM or about 70mM, or about 100mM.

The isotonic agent as used herein includes salts, e.g., sodium chloride, dextrose, lactose or combination thereof.

The preservative as used herein include, but are not limited to, benzoic acid, butylparaben, ethyl paraben, methyl paraben, propylparaben, sodium benzoate, sodium propionate, benzalkonium chloride, benzethonium chloride, benzyl alcohol, cetypyridinium chloride, chlorobutanol, phenol, phenylethyl alcohol, 2-penoxyethanol, phenyl mercuric nitrate, thimerosal, metacresol or combinations thereof. The concentration of preservative is about 0.1mg/mL to about 5mg/mL, or about 1mg/ml, or about 2 mg/ml or about 2.5 mg/mL or about 3 mg/mL.

The pH modifying agents as used herein refers to a combination of acid and alkali. The pH modifying agents can be selected from the group comprising of hydrochloric acid, o-phosphoric acid, citric acid, acetic acid, succinic acid, lactic acid, gluconic acid, tartaric acid, 1,2,3,4-butane tetracarboxylic acid, fumaric acid and malic acid. Alkali is selected from the group comprising of sodium hydroxide, potassium hydroxide, sodium hydroxide, ammonium hydroxide, magnesium oxide, calcium hydroxide, calcium carbonate, magnesium carbonate, magnesium aluminum silicates, diethanolamine, monoethanolamine, sodium carbonate, sodium bicarbonate, triethanolamine and combination thereof.

In another embodiment, the composition is stable for at least 24 hours when stored in glass vials at 25 °C and relative humidity of 60%. In another embodiment, the composition is stable for at least 6 months when stored in glass vials at 25 °C and relative humidity of 60%. In another embodiment, the stability studies were performed as per standard in-house method.

In yet another embodiment, the two amino acids, arginine and isoleucine are present in a weight ratio of about 1:1 to about 1:5, or about 1:2 to about 1:3, or about 1:3 to about 1:4, or about 1:4 to about 1:5, preferably about 1:2. In an embodiment, the concentration of arginine or isoleucine ranges from about 1mM to 100mM, or about 2mM to 50mM, or about 3mM to 25mM. In an embodiment, the concentration of arginine is about 2mM to 20mM, or about 2mM to 10mM. Alternatively, the concentration of arginine is about 3mM, or about 4mM, or about 5mM, or about 6mM, or about 7mM, or about 8mM, or about 9mM. In an embodiment, the concentration of isoleucine is about 2mM to 20mM, or about 2mM to 15mM. Alternatively, the concentration of isoleucine is about 3mM, or about 4mM, or about 5mM, or about 6mM, or about 7mM, or about 8mM, or about 9mM, or about 10mM, or about 11mM, or about 12mM, or about 13mM, or about 14mM. Alternatively, the concentration of arginine is about 7mM and the concentration of isoleucine is about 14mM.

The pharmaceutical composition of the present invention is suitable for parenteral administration such as intramuscular, subcutaneous, intradermal and intravenous administration. Preferably, the pharmaceutical composition is suitable for subcutaneous injection administration.

In another aspect of the invention, there is provided an aqueous pharmaceutical composition comprising: (a) from about 100 IU/ml of buffered insulin glargine, (b) therapeutically effective amount of liraglutide, (c) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 6 to about 9. In one embodiment, the buffered insulin glargine and liraglutide are present separated from each other or as a mixture. In another embodiment, the buffered insulin glargine and liraglutide are formulated as a fixed dose composition.

In one embodiment, there is provided an aqueous pharmaceutical composition comprising: (a) about 100 IU/ml of buffered insulin glargine, (b) about 0.6 mg/mL of liraglutide, and (c) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 3 to about 9.

In one embodiment, there is provided an aqueous pharmaceutical composition comprising: (a) about 100 IU/ml of buffered insulin glargine, (b) about 0.3 mg/mL of liraglutide, and (c) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 3 to about 9. In another embodiment, amount of liraglutide is about 0.6 mg/mL or about 1.2 mg/mL or about 1.8 mg/mL.

In yet another embodiment, there is provided an aqueous pharmaceutical composition comprising: (a) 3.64 mg of buffered insulin glargine, (b) about 0.3 mg of liraglutide, (c) 10 mg of 85% glycerol, (d) 30 µg of zinc Chloride, (e) 2.7 mg of m-cresol, (f) 5mM of L-arginine and 10 mM of isoleucine and (g) 1 mg of dipotassium phosphate, and wherein the pharmaceutical composition has a pH of 8.2 ± 0.2.

In yet another embodiment, there is provided an aqueous pharmaceutical composition comprising: (a) 3.64 mg of buffered insulin glargine, (b) about 0.6 mg of liraglutide, (c) 10 mg of 85% glycerol, (d) 30 µg of zinc Chloride, (e) 2.7 mg of m-cresol, (f) 5mM of L-arginine and 10 mM of isoleucine and (g) 1 mg of dipotassium phosphate, and wherein the pharmaceutical composition has a pH of 8.2 ± 0.2.

In another aspect, there is provided an aqueous pharmaceutical composition comprising: (a) liraglutide and buffered insulin glargine in a weight ratio of about 1:1 to about 1:20, and (b) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 6 to about 9. In another embodiment, liraglutide and buffered insulin glargine present in a weight ratio of about 1:2 to about 1:18, or about 1:3 to about 1:15, or about 1:6 to about 1:12, wherein the one or more pharmaceutically acceptable excipients comprise a pH maintaining agent, solubilising agent, isotonic agent, preservative, pH modifying agent or combination thereof.

In yet another embodiment, there is provided an aqueous pharmaceutical composition comprising: (a) about 100 IU/ml of buffered insulin glargine, (b) about 0.3 mg/mL to about 2 mg/mL of liraglutide, and (c) one or more pharmaceutically acceptable excipients selected from a pH maintaining agent, solubilising agent, isotonic agent, preservative, pH modifying agent or combination thereof, wherein the pharmaceutical composition has a pH between about 3 to about 9. In another embodiment, wherein the pharmaceutical composition comprises about 0.6 mg/mL of liraglutide, and wherein the pharmaceutical composition has a pH between about 6 to about 9. In another embodiment, wherein the pharmaceutical composition comprises about 1.2 mg/mL of liraglutide, and wherein the pharmaceutical composition has a pH between about 6 to about 9. In another embodiment, wherein the pharmaceutical composition comprises about 1.8 mg/mL of liraglutide, and wherein the pharmaceutical composition has a pH between about 6 to about 9.

In another aspect of this invention, there is provided an aqueous pharmaceutical composition comprising first component and second component, wherein said first component comprises (a) buffered insulin glargine, and (b) a pharmaceutically acceptable excipient, and said second component comprises (a) liraglutide, and (b) a pharmaceutically acceptable excipient. In one embodiment, the first component and second component can be administered as mixture or separately. In another embodiment, the composition is formulated as a fixed dose composition. In another embodiment, pharmaceutically acceptable excipients from first and second component are selected from a pH maintaining agent, solubilising agent, isotonic agent, preservative, pH modifying agent or combination thereof

In another aspect of the invention, there is provided a kit comprising an aqueous composition comprising first component and second component, wherein said first component comprises (a) buffered insulin glargine, and (b) a pharmaceutically acceptable excipient, and said second component comprises (a) liraglutide, and (b) a pharmaceutically acceptable excipient. In another embodiment, the composition is formulated as a fixed dose composition. In another embodiment, pharmaceutically acceptable excipients from first and second component are selected from a pH maintaining agent, solubilising agent, isotonic agent, preservative, pH modifying agent or combination thereof.

In another aspect of this invention, there is provided a method of treating diabetes mellitus in a subject, said method comprises parenterally administering to said subject an aqueous pharmaceutical composition comprising: (a) from about 1 IU/ml to about 500 IU/ml of buffered insulin glargine, (b) therapeutically effective amount of a GLP-1 analogue, and (c) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 3 to about 9. In an embodiment, said method comprises parenterally administering to said subject an aqueous pharmaceutical composition comprising: (a) liraglutide and buffered insulin glargine in a weight ratio of about 1:1 to about 1:20, and (b) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 6 to about 9.

In another aspect of this invention, there is provided a use of the aqueous composition of any aspect of this invention for the preparation of medicament for treating diabetes, for adjusting the fasting, postprandial and/or post absorptive blood glucose concentration, for improving glucose tolerance, for preventing hypoglycemia, for preventing loss of function of the pancreatic ß-cells and for weight loss and/or for preventing weight gain in a patient in need thereof.

In another aspect of this invention, there is provided a process of using the kit comprising aqueous composition comprising first component and second component, wherein said first component comprises (a) buffered insulin glargine, and (b) a pharmaceutically acceptable excipient, and said second component comprises (a) liraglutide, and (b) a pharmaceutically acceptable excipient, wherein the process comprises the steps of:
(i) selecting a dose of the buffered insulin glargine that is to be administered,
(ii) selecting a dose of the liraglutide that is to be administered,
(iii) determining and administering an amount which corresponds to the doses of buffered insulin glargine and liraglutide.

In another aspect of this invention, there is provided a method of preparing a pharmaceutical composition in the form of solution, said method comprising the steps of:
(i) preparing a solution comprising insulin glargine in acidic water for injection,
(ii) preparing a solution of at least two amino acids,
(iii) adding the solution of at least two amino acids in the solution of step (i) and adjusting the pH to about 3 to about 4 using a pH modifying agent,
(iv) preparing a solution comprising liraglutide, and
(v) mixing the solutions of step (iii) and (iv) and adjusting the pH to about 8.2± 0.2 to obtain the pharmaceutical composition.

In another embodiment, there is provided a method of preparing a pharmaceutical composition in the form of suspension, said method comprising the steps of:
(i) preparing a solution comprising insulin glargine in acidic water for injection,
(ii) preparing a solution of at least two amino acids,
(iii) adding the solution of at least two amino acids in the solution of step (i) and adjusting the pH to about 6 to about 9 using a pH modifying agent,
(iv) preparing a solution comprising liraglutide, and
(v) mixing the solutions of step (iii) and (iv) and adjusting the pH to about 8.2± 0.2 to obtain the pharmaceutical composition.

EXAMPLES
EXAMPLE 1: Aqueous composition of buffered glargine and liraglutide
Part A. Preparation of buffered insulin glargine
S. No. Ingredients Quantity/mL
1. Insulin Glargine 100IU (3.64mg)
2. Zinc Chloride 30 µg
3. L- Arginine 5.0mM
4. Isoleucine 10.0mM
5. Dipotassium phosphate 1 mg
6. Sodium Hydroxide Q.S.
7. Hydrochloric Acid Q.S.
8. Water for Injection Q.S. to 1.0 mL

Process: Zinc-containing crystals of insulin glargine were dissolved in water for injection with the help of 1M HCl. To this solution dipotassium phosphate, L-arginine and isoleucine in water for injection were added to get final concentration. The pH was adjusted to 4.0±0.2 with 1M HCl or 1M NaOH. The solution was filtered with 0.2 micron filter in a sterile container.
Part B. Preparation of liraglutide solution
S. No. Ingredients Quantity/mL
1. Liraglutide 0.6 mg
2. Glycerol 10 mg
3. m-cresol 2.7 mg
4. Dipotassium phosphate 1 mg
5. Sodium Hydroxide Q.S.
6. Hydrochloric Acid Q.S.
7. Water for Injection Q.S. to 1.0 mL
Process: Glycerol, m-cresol and dipotassium phosphate were dissolved in water for injection. To this solution liraglutide was added and the final volume was made. The pH was adjusted to 8.0±0.2 with 1M HCl or 1M NaOH. The solution was filtered with 0.2 micron filter in a sterile container.
Part C: Preparation of composition of buffered glargine and liraglutide
The solution of part B was added slowly to solution of part A, mixed thoroughly and adjusted the volume. The pH was adjusted to 8.2±0.2.

EXAMPLE 2: Fixed dose compositions of buffered insulin glargine and liraglutide
S. No. Ingredients Compositions
Quantity/mL
A B C D
1. Insulin Glargine 100IU
(3.64 mg) 150 IU
(5.46 mg) 200 IU
(7.28 mg) 300 IU
(10.92 mg)
2. Liraglutide 0.3 mg 0.6 mg 1.2 mg 1.8 mg
3. 85% glycerol 10 mg 11 mg 12 mg 15 mg
4. Zinc Chloride 30 µg 32 µg 35µg 40 µg
5. m-cresol 2.7 mg 2.8 mg 2.9 mg 3 mg
6. L- Arginine 5 mM 10 mM 15 mM 20 mM
7. Isoleucine 10.0 mM 15 mM 25 mM 40 mM
8. Dipotassium phosphate 1 mg 1.5 mg 2 mg 4 mg
9. Sodium Hydroxide Q.S. Q.S. Q.S. Q.S.
10. Hydrochloric Acid Q.S. Q.S. Q.S. Q.S.
11. Water for Injection Q.S. to 1.0 mL Q.S. to 1.0 mL Q.S. to 1.0 mL Q.S. to 1.0 mL

Manufacturing process:
1. Zinc-containing crystals of insulin glargine were dissolved in water for injection with the help of 1M HCl. The endogenous zinc level was supplemented by adding appropriate volume of zinc chloride solution (1% w/v). To this L-arginine and isoleucine was added with mixing. The pH was adjusted to 4.0±0.2 with 1M HCl or 1M NaOH.
2. A solution was prepared by dissolving m-cresol, glycerol and dipotassium phosphate in water for injection to get final concentration.
3. Liraglutide was dissolved in the solution of step 2 and pH was adjusted to 8.0±0.2 with 1M HCl or 1M NaOH.
4. Both solution of step 1 and solution of step 3 were diluted to final concentration after mixing and pH was adjusted to 8.2±0.2 with 1M HCl or 1M NaOH.

EXAMPLE 3: Fixed dose compositions of buffered insulin glargine and liraglutide
S. No. Ingredients Compositions
Quantity/mL
A B C D
1. Insulin Glargine 100IU
(3.64 mg) 100 IU
(3.64 mg) 100 IU
(3.64 mg) 100 IU
(3.64 mg)
2. Liraglutide 0.3 mg 0.6 mg 1.2 mg 1.8 mg
3. 85% glycerol 10 mg 10 mg 10 mg 10 mg
4. Zinc Chloride 30 µg 30 µg 30 µg 30 µg
5. m-cresol 2.7 mg 2.7 mg 2.7 mg 2.7 mg
6. L- Arginine 5 mM 5 mM 5 mM 5 mM
7. Isoleucine 10.0 mM 10.0 mM 10.0 mM 10.0 mM
8. Dipotassium phosphate 1 mg 1 mg 1 mg 1 mg
9. Sodium Hydroxide Q.S. Q.S. Q.S. Q.S.
10. Hydrochloric Acid Q.S. Q.S. Q.S. Q.S.
11. Water for Injection Q.S. to 1.0 mL Q.S. to 1.0 mL Q.S. to 1.0 mL Q.S. to 1.0 mL
Manufacturing process: Manufacturing process is same as mentioned in Example 2.
EXAMPLE 4: Stability Studies of compositions of buffered insulin glargine and liraglutide
A. Stability data of Example 1, and Examples 3A to 3D stored at 25°C and relative humidity of 60% for 24 Hours
Compositions Time in Hours High Molecular weight impurities Other Impurities



Example 1 Initial 0.01 0.09
2 0.01 0.14
4 0.03 0.29
8 0.04 0.53
24 0.12 0.86
Example 3A Initial 0.01 0.16
2 0.02 0.34
4 0.05 0.74
8 0.06 0.58
24 0.15 0.90
Example 3B Initial 0.01 0.09
2 0.01 0.14
4 0.03 0.29
8 0.04 0.53
24 0.12 0.86
Example 3C Initial 0.01 0.1
2 0.03 0.19
4 0.07 0.36
8 0.06 0.62
24 0.21 0.95
Example 3D Initial 0.01 0.16
2 0.05 0.21
4 0.06 0.46
8 0.09 0.71
24 0.19 0.87

B. Stability data of Example 1, and Examples 3A to 3D stored at 25°C and relative humidity of 60% for 6 months

Compositions Time in Months High Molecular weight impurities Other Impurities



Example 1 Initial 0.01 0.03
1 0.05 0.05
2 0.09 0.07
4 0.13 0.11
6 0.18 0.19
Example 3A Initial 0.01 0.05
1 0.12 0.15
2 0.23 0.42
4 0.35 0.89
6 0.42 0.64
Example 3B Initial 0.01 0.03
1 0.05 0.05
2 0.09 0.07
4 0.13 0.11
6 0.18 0.19
Example 3C Initial 0.01 0.05
1 0.1 0.11
2 0.23 0.21
4 0.4 0.46
6 0.49 0.73
Example 3D Initial 0.01 0.07
1 0.05 0.21
2 0.18 0.36
4 0.36 0.52
6 0.65 0.78

,CLAIMS:1. An aqueous pharmaceutical composition comprising: (a) from about 1 IU/ml to about 500 IU/ml of buffered insulin glargine, (b) therapeutically effective amount of a GLP-1 analogue, and (c) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 3 to about 9.

2. An aqueous pharmaceutical composition comprising: (a) liraglutide and buffered insulin glargine in a weight ratio of about 1:1 to about 1:20, and (b) one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition has a pH between about 6 to about 9.

3. The pharmaceutical composition of claim 2, wherein the liraglutide to the buffered insulin glargine weight ratio is about 1:2 to about 1:18.

4. The pharmaceutical composition of claim 2, the liraglutide to the buffered insulin glargine weight ratio is about 1:3 to about 1:15.

5. The pharmaceutical composition of claim 2, the liraglutide to the buffered insulin glargine weight ratio is about 1:6 to about 1:12.

6. The pharmaceutical composition of claim 1, wherein the GLP-1 analogue comprises exenatide, liraglutide, lixisenatide, semaglutide, dulaglutide and albiglutide.

7. The pharmaceutical composition of claim 6, wherein the GLP-1 analogue is liraglutide, present in the concentration from about 0.3 mg/ml to about 2 mg/ml.

8. The pharmaceutical composition of any one of claims 1-2, wherein the buffered insulin glargine is present in an amount of about 40IU/mL to about 500IU/mL.

9. The pharmaceutical composition of any one of claims 1-2, wherein the one or more pharmaceutically acceptable excipients comprise a pH maintaining agent, solubilising agent, isotonic agent, preservative, pH modifying agent or combination thereof.

10. An aqueous pharmaceutical composition comprising: (a) about 100 IU/ml to about 300 IU/ml of buffered insulin glargine, (b) about 0.3 mg/mL to about 2 mg/mL of liraglutide, and (c) one or more pharmaceutically acceptable excipients selected from a pH maintaining agent, solubilising agent, isotonic agent, preservative, pH modifying agent or combination thereof, wherein the pharmaceutical composition has a pH between about 3 to about 9.