DESC:BACKGROUND OF THE INVENTION
Diabetes is a disease characterized by abnormally high levels of blood glucose and inadequate level of insulin. There are two types of diabetes- Type I and Type II diabetes. In Type I diabetes, also known as Diabetes Insipidus or Insulin dependent diabetes mellitus (IDDM), body produces no insulin. Insulin is a peptide hormone, produced by beta cells in the pancreas and is central to regulating carbohydrate and fat metabolism in the body. Since the introduction of insulin in the 1920’s, continuous efforts have been made to improve the treatment of diabetes mellitus. To avoid extreme glycemic levels, diabetic patients often practice multiple injection therapy, whereby insulin is administered with each meal. A number of insulin, insulin analogues and derivatives are available in the market. The commonly used types of insulin, insulin analogues or insulin derivatives are categorized as: (1) Rapid-acting Insulin analogs (Bolus): For example insulin aspart (Novolog®); insulin lispro (Humalog®); insulin glulisine (Aprida®), rapid acting human insulin (Viaject®); (2) Short-acting insulin (Bolus): For example regular insulin (Humulin® ar Novolin®); (3) Intermediate-acting insulin: For example as isophane insulin, aspart protamine, lispro protamine; (4) Long-acting insulin (Basal): For example insulin glargine, insulin degludec and insulin detemir; (5) Mixed Insulin: For example mixture of NPH and regular insulin.

When a patient takes food, there is a steep rise in the blood glucose level but due to the initial lag phase, basal insulin analogues are not able to control the initial spike in the post prandial blood glucose level. Therefore, doctors prescribe rapid insulin analogues injections along with the basal insulin analogues. As basal insulin analogues cannot be pre-mixed with rapid insulin analogues, basal insulin analogues and rapid insulin analogues are given in separate injections to the patient. With three meals a day, a patient needs to take three injections of rapid insulin analogues and one or two injections of basal insulin analogue. With four injections a day, hundred twenty injections a month and one thousand four hundred and forty injections in a year, repeated administration of insulin or insulin analogues separately many times a day is a major cause of patient non-compliance.

US Patent No. 8,575,096 discloses fast acting insulin analogues which can form soluble mixtures (pre-mixed or self-mixed) with long acting insulin analogues.
US Patent No. 8,084,420 discloses an injectable formulation containing a rapid acting insulin and a long acting insulin, wherein the pH of the fast or rapid acting insulin is adjusted so that both rapid and long acting insulin’s remain soluble when they are mixed together. US Patent No. 7,713,930 discloses a pharmaceutical formulation comprising insulin, insulin metabolite, insulin analogue, an insulin derivative and combinations thereof and pharmaceutically acceptable excipients. US Patent Application No. 2015/0126439A1 discloses a combination comprising at least a first insulin-like compound and a second insulin-like compound for treating a condition or a disease in a subject in need thereof. US Patent No. 7,476,652 discloses a pharmaceutical formulation comprising a polypeptide selected from the group consisting of insulin, an insulin metabolite, an insulin analog, an insulin derivative and combinations thereof, preservative and water, wherein the pharmaceutical formulation has a pH in the acidic range from 1 to 6.8. PCT Publication No. WO2015/044922 A1 discloses readily dissociable molecular aggregates formed by combining an insulin, insulin analogue, derivative or metabolite having isoelectric point between 5.8 to 8.5 in combination with one or more an insulin, insulin analogue, derivative or metabolite having isoelectric point between 4.0 to 5.7, wherein the said molecular aggregates have an average particle size between about 5 µm to about 20µm.

The most common problem with the mixing of basal insulin analogues and rapid insulin analogues is the precipitation of basal insulin analogues due to difference in their isoelectric points. Insulin glargine, a basal insulin analogue has an isoelectric point (pI) between 5.8 to 8.5 and thus it remains in solution form at a pH below 4.0. On the other hand the pI of rapid insulin analogues such as insulin aspart and insulin lispro is less than 5.7 and thus the formulations comprising rapid insulin analogues have neutral pH 7.0 to 7.4. Mixing of pharmaceutical compositions of insulin glargine (pH 3.8-4.2) and rapid acting insulin analogue (pH 7.2-7.4) leads to formation insulin glargine precipitates on which rapid acting insulin analogue gets absorbed. The formation of precipitates in the formulation during delivery or storage is unacceptable. Further, at acidic pH rapid acting insulin are soluble but not stable.

There is a need to develop a pharmaceutical composition comprising insulin, insulin analogue, insulin metabolite or derivative thereof, which does not precipitate and provide adequate stability on mixing with formulation of basal insulin analogue in a single injection.

SUMMARY OF THE INVENTION
In general aspect of this invention, there is provided a ready-to-use pharmaceutical composition comprising: (a) insulin glargine, (b) Gly(A21), Lys(B28), Pro(B29) human insulin, (c) 0.2mg/ml to 10mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In first aspect of the invention, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5. In one embodiment of this aspect, the insulin glargine is present in a concentration of about 70IU/mL and the Gly(A21), Lys(B28), Pro(B29) human insulin is present in the concentration of about 30IU/mL. In another embodiment of this aspect, the pharmaceutical composition further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition. In still another embodiment, the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof. In still another embodiment, the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition. In still another embodiment, the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof. In still another embodiment, the surfactant is present in the concentration from about 5mM to about 100mM.

In one embodiment, there is provided a stable ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In second aspect of this invention, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, (d) about 0.1 mg/mL to about 2 mg/mL of protamine salt, and (e) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5. In one embodiment of this aspect, the insulin glargine is present in a concentration of about 70IU/mL and the Gly(A21), Lys(B28), Pro(B29) human insulin is present in the concentration of about 30IU/mL. In another embodiment of this aspect, the protamine salt is present in the concentration of about 0.5mg/mL. In still another embodiment, the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof. In still another embodiment, the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition. In still another embodiment, the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof. In still another embodiment, the surfactant is present in the concentration from about 5mM to about 100mM.

In one embodiment, there is provided a stable ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, (d) about 0.1 mg/mL to about 2 mg/mL of protamine salt, and (e) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In third aspect of this invention, there is provided a process of preparing a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the process comprises steps of:
a) preparing an aqueous solution comprising insulin glargine and glycine having the pH of about 3.5 to about 4.5,
b) preparing an aqueous solution comprising Gly(A21), Lys(B28), Pro(B29) human insulin having the pH of about 7.2 to about 8.0, and
c) adding the solution of step (b) in the solution of step (a) such that the final pH of the composition is in the range of about 3.5 to about 4.5.

In fourth aspect of this invention, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, (d) about 0.5 mg/mL of protamine salt, and (e) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In fifth aspect of this invention, there is provided ready-to-use pharmaceutical composition comprising: (a) about 70IU/mL of insulin glargine, (b) about 30IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In sixth aspect of this invention, there is provided a kit comprising a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5. In one embodiment of this aspect, the insulin glargine is present in a concentration of about 70IU/mL and the Gly(A21), Lys(B28), Pro(B29) human insulin is present in the concentration of about 30IU/mL. In another embodiment of this aspect, the pharmaceutical composition further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition. In still another embodiment, the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof. In still another embodiment, the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition. In still another embodiment, the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof. In still another embodiment, the surfactant is present in the concentration from about 5mM to about 100mM.

In last aspect of this invention, there is provided use of a ready-to-use pharmaceutical composition for treatment of type I or type II diabetes, said composition comprises: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5. In one embodiment of this aspect, the insulin glargine is present in a concentration of about 70IU/mL and the Gly(A21), Lys(B28), Pro(B29) human insulin is present in the concentration of about 30IU/mL. In another embodiment of this aspect, the pharmaceutical composition further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition. In still another embodiment, the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof. In still another embodiment, the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition. In still another embodiment, the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof. In still another embodiment, the surfactant is present in the concentration from about 5mM to about 100mM.

DETAILED DESCRIPTION OF THE INVENTION
In general aspect of this invention, there is provided a ready-to-use pharmaceutical composition comprising: (a) insulin glargine, (b) Gly(A21), Lys(B28), Pro(B29) human insulin, (c) 0.2mg/ml to 10mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5. In one embodiment, there is provided a stable ready-to-use pharmaceutical composition comprising: (a) insulin glargine, (b) Gly(A21), Lys(B28), Pro(B29) human insulin, (c) 0.2mg/ml to 10mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

For the clarity, insulin glargine in the present invention has the amino acid sequence as:
A chain : GIVEQCCTSICSLYQLENYCG
B chain : FVNQHLCGSHLVEALYLVCGERGFFYTPKTRR
For the clarity, Gly(A21), Lys(B28), Pro(B29) human insulin of the invention composition has the amino acid sequence as:
A chain : GIVEQCCTSICSLYQLENYCG
B chain : FVNQHLCGSHLVEALYLVCGERGFFYTKPT

In one general embodiment, the invention compositions are for parenteral administration i.e., by intramuscular, subcutaneous, intravenous or intradermal route.

In one general embodiment, the invention compositions are formulated as solution, suspension or in the form of a lyophilized powder ready for reconstitution.

Ready-to-use composition relates to a premixed fixed dose composition of insulin glargine and Gly(A21), Lys(B28), Pro(B29) human insulin and can be administered directly to patient in a required dosage.

The invention compositions are physical and chemically stable on long storage i.e., 12 to 24 months. The amount of impurity generation (high molecular weight impurities and related impurities) is relatively lower in invention composition on long storage. Further, the invention compositions remain clear and colorless over a long period of time i.e., 12 to 24 months.

In first aspect of the invention, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In one embodiment, there is provided a stable ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In one embodiment, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL of insulin glargine, (b) about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In one embodiment, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 80IU/mL of insulin glargine, (b) about 20IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In one embodiment, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 70IU/mL of insulin glargine, (b) about 30IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In another embodiment, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5; and wherein the pharmaceutical composition further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition; and wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof; and wherein the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition; and wherein the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof; and wherein the surfactant is present in the concentration from about 5mM to about 100mM.

In another embodiment, there is provided a stable ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5; and wherein the pharmaceutical composition further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition; and wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof; and wherein the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition; and wherein the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof; and wherein the surfactant is present in the concentration from about 5mM to about 100mM.

In another embodiment, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL of insulin glargine, (b) about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5; and wherein the pharmaceutical composition further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition; and wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof; and wherein the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition; and wherein the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof; and wherein the surfactant is present in the concentration from about 5mM to about 100mM.

In another embodiment, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 80IU/mL of insulin glargine, (b) about 20IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5; and wherein the pharmaceutical composition further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition; and wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof; and wherein the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition; and wherein the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof; and wherein the surfactant is present in the concentration from about 5mM to about 100mM.

In another embodiment, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 70IU/mL of insulin glargine, (b) about 30IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5; and wherein the pharmaceutical composition further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition; and wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof; and wherein the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition; and wherein the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof; and wherein the surfactant is present in the concentration from about 5mM to about 100mM.

In another embodiment, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5; and wherein the pharmaceutical composition further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition; and wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof; and wherein the solubilizing agent is trehalose present in the concentration ranging from about 1.5% by weight of composition; and wherein the surfactant is n-octyl-ß-d-glucopyranoside present in about 10mM and n-dodecyl-ß-d-maltoside present in about 15mM.

In one embodiment, the concentration of insulin glargine or Gly(A21), Lys(B28), Pro(B29) human insulin ranges from about 10IU/mL to about 500IU/mL. The proportions between insulin glargine and Gly(A21), Lys(B28), Pro(B29) human insulin are, for example, as percentage, 25/75, 30/70, 40/60, 50/50, 60/40, 70/30, 80/20 or 90/10 for the invention compositions comprising about 10IU/mL to about 500IU/mL of insulin glargine or Gly(A21), Lys(B28), Pro(B29) human insulin.

In one embodiment, the invention composition comprises glycine in the concentration range of 0.2mg/mL to about 10mg/mL. Alternatively, the concentration of glycine ranges from about 0.4mg/mL to about 8mg/mL, or 0.6mg/mL to about 6mg/mL, or 0.8mg/mL to about 4mg/mL, or 1mg/mL to about 2mg/mL. Alternatively, the concentration of glycine is about 0.3mg/mL, or about 0.4mg/mL, or about 0.5mg/mL, or about 0.6g/mL, or about 0.7mg/mL, or about 0.8mg/mL, or about 0.9mg/mL, or about 1mg/mL, or about 1.5mg/mL, or about 2mg/mL, or about 4mg/mL, or about 6mg/mL, or about 8mg/mL.

In another embodiment, the invention composition comprises pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof.

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 or malic acid. Alkali is selected form 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 or triethanolamine and combination thereof.

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, trehalose 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), glycerol, 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 (preferred is Trehalose) is ranges from about 0.5% to about 20% by weight of composition. Alternatively, the concentration of solubilizing agent is about 1%, or about 1.5%, or about 2%, or about 3%, or about 4%, or about 6%, or about 8 %, or about 12%, or about 14%, or about 16%, or about 18% by weight of composition.

The surfactant as used herein includes, but is not limited to, n-octyl-ß-d-glucopyranoside, n-dodecyl-ß-d-maltoside or combination thereof. The concentration of n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside is about 5mM to about 100mM. The preferred concentration of n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof is about 10mM, or 15mM, or 20mM, or 25mM, or 30mM, or 35mM, or 40mM, or 45mM, or 50mM, or 60mM, or 70mM, or 80mM, or 90mM.

The isotonic agent as used herein include salts, e.g., sodium chloride, dextrose, lactose and 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 and combinations thereof. The concentration of preservative is about 1mg/mL to about 5mg/mL. Preferred concentration of preservative is about 2.7 mg/mL.

The antioxidant as used herein can be selected from the group comprising of ascorbate (sodium/acid), bisulite sodium, butylated hydroxy anisole (BHA), butylated hydroxy toluene (BHT), cysteine / cysteinate HCl, dithionite sodium, gentisic acid, gentisic acid ethanolamine, glutamate monosodium, glutathione, formaldehyde sulfoxylate sodium, metabisulite potassium, metabisulite sodium, methionine, monothioglycerol (thioglycerol), propyl gallate, sulfite sodium, tocopherol alpha, alpha tocopherol hydrogen succinate, thioglycolate sodium and combination thereof.
Solvent used in the invention composition is water for injection.

In second aspect of this invention, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, (d) about 0.1 mg/mL to about 2 mg/mL of protamine salt, and (e) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In one embodiment, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL of insulin glargine, (b) about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, (d) about 0.1 mg/mL to about 2 mg/mL of protamine salt, and (e) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5; and wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof; and wherein the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition; and wherein the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof; and wherein the surfactant is present in the concentration from about 5mM to about 100mM.

In one embodiment, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 70IU/mL of insulin glargine, (b) about 30IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, (d) about 0.1 mg/mL to about 2 mg/mL of protamine salt, and (e) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5; and wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof; and wherein the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition; and wherein the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof; and wherein the surfactant is present in the concentration from about 5mM to about 100mM.

In one embodiment, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 80IU/mL of insulin glargine, (b) about 20IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, (d) about 0.1 mg/mL to about 2 mg/mL of protamine salt, and (e) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5; and wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof; and wherein the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition; and wherein the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof; and wherein the surfactant is present in the concentration from about 5mM to about 100mM.

In one embodiment, the protamine salt is protamine sulphate and is present in the concentration of about 0.1 mg/mL to about 2 mg/mL. Alternatively, the concentration of protamine salt is about 0.2 mg/mL, about 0.3 mg/mL, or about 0.4 mg/mL, about 0.5 mg/mL, or about 0.6 mg/mL, about 0.7 mg/mL, or about 0.8 mg/mL, or about 1 mg/mL, or about 1.2 mg/mL, or about 1.4 mg/mL, or about 1.6 mg/mL, or about 1.8 mg/mL.

In third aspect of this invention, there is provided a process of preparing a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the process comprises steps of:
a) preparing an aqueous solution comprising insulin glargine and glycine having the pH of about 3.5 to about 4.5,
b) preparing an aqueous solution comprising Gly(A21), Lys(B28), Pro(B29) human insulin having the pH of about 7.2 to about 8.0, and
c) adding the solution of step (b) in the solution of step (a) such that the final pH of the composition is in the range of about 3.5 to about 4.5.

In one embodiment, there is provided a process of preparing a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL of insulin glargine, (b) about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the process comprises steps of:
a) preparing an aqueous solution comprising insulin glargine and glycine having the pH of about 3.5 to about 4.5,
b) preparing an aqueous solution comprising Gly(A21), Lys(B28), Pro(B29) human insulin having the pH of about 7.2 to about 8.0, and
c) adding the solution of step (b) in the solution of step (a) such that the final pH of the composition is in the range of about 3.5 to about 4.5.

In another embodiment, there is provided a process of preparing a ready-to-use pharmaceutical composition comprising: (a) about 70IU/mL of insulin glargine, (b) about 30IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and the process comprises steps of:
a) preparing an aqueous solution comprising insulin glargine and glycine having the pH of about 3.5 to about 4.5,
b) preparing an aqueous solution comprising Gly(A21), Lys(B28), Pro(B29) human insulin having the pH of about 7.2 to about 8.0, and
c) adding the solution of step (b) in the solution of step (a) such that the final pH of the composition is in the range of about 3.5 to about 4.5.

In another embodiment, there is provided a process of preparing a ready-to-use pharmaceutical composition comprising: (a) about 80IU/mL of insulin glargine, (b) about 20IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the process comprises steps of:
a) preparing an aqueous solution comprising insulin glargine and glycine having the pH of about 3.5 to about 4.5,
b) preparing an aqueous solution comprising Gly(A21), Lys(B28), Pro(B29) human insulin having the pH of about 7.2 to about 8.0, and
c) adding the solution of step (b) in the solution of step (a) such that the final pH of the composition is in the range of about 3.5 to about 4.5.

In fourth aspect of this invention, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, (d) about 0.5 mg/mL of protamine salt, and (e) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In one embodiment, there is provided a stable ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, (d) about 0.5 mg/mL of protamine salt, and (e) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In one embodiment, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL of insulin glargine, (b) about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, (d) about 0.5 mg/mL of protamine salt, and (e) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In one embodiment, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 70IU/mL of insulin glargine, (b) about 30IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, (d) about 0.5 mg/mL of protamine salt, and (e) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In one embodiment, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 80IU/mL of insulin glargine, (b) about 20IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, (d) about 0.5 mg/mL of protamine salt, and (e) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In fifth aspect of this invention, there is provided a ready-to-use pharmaceutical composition comprising: (a) about 70IU/mL of insulin glargine, (b) about 30IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In one embodiment, there is provided a stable ready-to-use pharmaceutical composition comprising: (a) about 70IU/mL of insulin glargine, (b) about 30IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In sixth aspect of this invention, there is provided a kit comprising a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In one embodiment, there is provided a kit comprising a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5; and wherein the pharmaceutical composition further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition; and wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof; and wherein the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition; and wherein the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof; and wherein the surfactant is present in the concentration from about 5mM to about 100mM.

In one embodiment, there is provided a kit comprising a ready-to-use pharmaceutical composition comprising: (a) about 70IU/mL of insulin glargine, (b) about 30IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5; and wherein the pharmaceutical composition further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition; and wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof; and wherein the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition; and wherein the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof; and wherein the surfactant is present in the concentration from about 5mM to about 100mM.

In last aspect of this invention, there is provided use of a ready-to-use pharmaceutical composition for treatment of type I or type II diabetes, said composition comprises: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In one embodiment, there is provided use of a stable ready-to-use pharmaceutical composition for treatment of type I or type II diabetes, said composition comprises: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

In one embodiment, there is provided use of a ready-to-use pharmaceutical composition for treatment of type I or type II diabetes, said composition comprises: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5; and wherein the pharmaceutical composition further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition; and wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof; and wherein the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition; and wherein the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof; and wherein the surfactant is present in the concentration from about 5mM to about 100mM.

In one embodiment, there is provided use of a ready-to-use pharmaceutical composition for treatment of type I or type II diabetes, said composition comprises: (a) about 60IU/mL of insulin glargine, (b) about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5; and wherein the pharmaceutical composition further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition; and wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof; and wherein the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition; and wherein the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof; and wherein the surfactant is present in the concentration from about 5mM to about 100mM.

In one embodiment, there is provided use of a ready-to-use pharmaceutical composition for treatment of type I or type II diabetes, said composition comprises: (a) about 70IU/mL of insulin glargine, (b) about 30IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5; and wherein the pharmaceutical composition further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition; and wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof; and wherein the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition; and wherein the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof; and wherein the surfactant is present in the concentration from about 5mM to about 100mM.

In one embodiment, there is provided use of a ready-to-use pharmaceutical composition for treatment of type I or type II diabetes, said composition comprises: (a) about 80IU/mL of insulin glargine, (b) about 20IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5; and wherein the pharmaceutical composition further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition; and wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof; and wherein the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of composition; and wherein the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof; and wherein the surfactant is present in the concentration from about 5mM to about 100mM.

While the invention has been described in term of its specific embodiments, certain modification and equivalents will be apparent to those skilled in the art and are intended to be included within the scope of the invention.

EXAMPLES
EXAMPLE 1: Pharmaceutical compositions of insulin glargine and Gly(A21), Lys(B28), Pro(B29) human insulin
S.No. Ingredients Composition
1A 1B 1C
1 Insulin Glargine (r-DNA origin) 70 IU/mL 60 IU/mL 80 IU/mL
2 Gly(A21), Lys(B28), Pro(B29) human insulin (r-DNA origin) 30 IU/mL 40 IU/mL 20 IU/mL
3 Glycine 1 mg/mL 1 mg/mL 1 mg/mL
4 Glycerol, 85% 20 mg/mL 20 mg/mL 20 mg/mL
5 Zinc oxide 0.30 mg/mL 0.30 mg/mL 0.30 mg/mL
6 m-cresol 2.7 mg/mL 2.7 mg/mL 2.7 mg/mL
7 Trehalose 1.5 % 1.5 % 1.5 %
8 n-octyl-ß-d-glucopyranoside 10mM 10mM 10mM
9 n-dodecyl-ß-d-maltoside 15mM 15mM 15mM
11 Sodium Hydroxide Q.S. to adjust pH Q.S. to adjust pH Q.S. to adjust pH
12 Hydrochloric acid Q.S. to adjust pH Q.S. to adjust pH Q.S. to adjust pH
13 Water for injection Q.S. to 1.0 mL Q.S. to 1.0 mL Q.S. to 1.0 mL
pH 4.0 ± 0.5 4.0 ± 0.5 4.0 ± 0.5

Manufacturing Procedure:
A. Preparation of solution of insulin glargine (100 IU)
Step 1: Preparation of insulin glargine Solution
Insulin glargine 100IU solution was prepared by dissolving accurately weighed zinc-containing crystals of insulin glargine in Water for Injection with the help of 1M HCl. The pH of this solution was adjusted to 4.0 ± 0.5 with 1M NaOH.
Step 2: Preparation of stabilizer solution
Stabilizer solution was prepared by dissolving the glycine, trehalose, n-octyl-ß-d-glucopyranoside, n-dodecyl-ß-d-maltoside and other excipients to get final concentration as per unit composition formula. The pH of this solution was adjusted to 4.0 ± 0.5 with 1M NaOH.
Step 3: Preparation of final solutions of desired concentrations
Both insulin glargine solution of step 1 and stabilizer solution of step 2 were mixed and pH was further adjusted to 4.0 ± 0.5.

B: Preparation of Gly(A21), Lys(B28), Pro(B29) human insulin solution (100IU)
Gly(A21), Lys(B28), Pro(B29) human insulin 100IU solution was prepared by dissolving accurately weighed zinc-containing crystals of Gly(A21), Lys(B28), Pro(B29) human insulin in water for Injection with 1M HCl. The pH of this solution was adjusted to 7.2-8.0 with 1M NaOH.

C: Mixture of insulin glargine and Gly(A21), Lys(B28), Pro(B29) human insulin
The solution of Gly(A21), Lys(B28), Pro(B29) human insulin was added slowly to the insulin glargine solution so as to obtain the different proportions of insulin glargine and Gly(A21), Lys(B28), Pro(B29) human insulin i.e., 70:30, 60:40 and 80:20. The solution was mixed thoroughly. The pH of the solution was adjusted to 4.0 ± 0.5.

EXAMPLE 2: Pharmaceutical compositions of insulin glargine and Gly(A21), Lys(B28), Pro(B29) human insulin (70/30)
S.No. Ingredients Composition
2A 2B 2C
1 Insulin Glargine (r-DNA origin) 70 IU/mL 70 IU/mL 70 IU/mL
2 Gly(A21), Lys(B28), Pro(B29) human insulin (r-DNA origin) 30 IU/mL 30 IU/mL 30 IU/mL
3 Glycine 1 mg/mL 1.5 mg/mL 2 mg/mL
4 Glycerol, 85% 20 mg/mL 20 mg/mL 20 mg/mL
5 Zinc oxide 0.30 mg/mL 0.30 mg/mL 0.30 mg/mL
6 m-cresol 2.7 mg/mL 2.7 mg/mL 2.7 mg/mL
7 Trehalose 1.5 % 1 % 2 %
8 n-octyl-ß-d-glucopyranoside 10mM 15mM 20mM
9 n-dodecyl-ß-d-maltoside 15mM 20mM 25mM
11 Sodium Hydroxide Q.S. to adjust pH Q.S. to adjust pH Q.S. to adjust pH
12 Hydrochloric acid Q.S. to adjust pH Q.S. to adjust pH Q.S. to adjust pH
13 Water for injection Q.S. to 1.0 mL Q.S. to 1.0 mL Q.S. to 1.0 mL
pH 4.0 ± 0.5 4.0 ± 0.5 4.0 ± 0.5

Manufacturing Procedure: The pharmaceutical compositions presented in example 2A to 2C were prepared by a similar process as described for examples 1A to 1C.
,CLAIMS:1. A ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; and wherein the composition has pH of about 3.5 to about 4.5.

2. The pharmaceutical composition of claim 1, wherein the insulin glargine is present in a concentration of about 70IU/mL and the Gly(A21), Lys(B28), Pro(B29) human insulin is present in the concentration of about 30IU/mL.

3. The pharmaceutical composition of claim 1, further comprises protamine salt present in the concentration of about 0.1mg/mL to about 2mg/mL by weight of composition.

4. The pharmaceutical composition of claim 1, wherein the pharmaceutically acceptable excipient comprises pH modifying agent, solubilizing agent, surfactant, isotonic agent, preservative, antioxidant or mixture thereof.

5. The pharmaceutical composition of claim 4, wherein the solubilizing agent is trehalose present in the concentration ranging from about 0.5% to about 20% by weight of the composition.

6. The pharmaceutical composition of claim 4, wherein the surfactant is n-octyl-ß-d-glucopyranoside or n-dodecyl-ß-d-maltoside or combination thereof.

7. The pharmaceutical composition of claim 4, wherein the surfactant is present in the concentration from about 5mM to about 100mM.

8. A process of preparing a ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient, wherein the process comprises steps of:
a) preparing an aqueous solution comprising insulin glargine and glycine having the pH of about 3.5 to about 4.5,
b) preparing an aqueous solution comprising Gly(A21), Lys(B28), Pro(B29) human insulin having the pH of about 7.2 to about 8.0, and
c) adding the solution of step (b) in the solution of step (a) such that the final pH of the composition is in the range of about 3.5 to about 4.5.

9. A ready-to-use pharmaceutical composition comprising: (a) about 60IU/mL to about 80IU/mL of insulin glargine, (b) about 20IU/mL to about 40IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, (d) about 0.5 mg/mL of protamine salt, and (e) a pharmaceutically acceptable excipient; wherein the composition has pH of about 3.5 to about 4.5.

10. A ready-to-use pharmaceutical composition comprising: (a) about 70IU/mL of insulin glargine, (b) about 30IU/mL of Gly(A21), Lys(B28), Pro(B29) human insulin, (c) about 1mg/mL of glycine, and (d) a pharmaceutically acceptable excipient; wherein the composition has pH of about 3.5 to about 4.5.