DESC:FORM 2

THE PATENTS ACT, 1970
(39 of 1970)
&
The Patents Rules, 2003

COMPLETE SPECIFICATION
(Section 10 and Rule 13)

LIRAGLUTIDE FORMULATIONS FOR PARENTERAL USE

AUROBINDO PHARMA LTD HAVING CORPORATE OFFICE AT
THE WATER MARK BUILDING,
PLOT NO.11, SURVEY NO.9,
KONDAPUR, HITECH CITY,
HYDERABAD - 500 084,
TELANGANA, INDIA
AN INDIAN ORGANIZATION

The following specification particularly describes the invention and the manner in which it is to be performed:

FIELD OF THE INVENTION

The present invention relates to a stable injectable formulation comprising a biologically active peptide Liraglutide, an isotonic agent, a buffering agent, and a preservative. The present invention also relates to processes for preparation of such stable formulation of Liraglutide with reduced deposits on production equipment and in the final product. The invention further relates to pharmaceutical formulations for subcutaneous administration of multiple doses with reduced clogging of injecting device. Furthermore the present invention is related to use of such formulations in the treatment of diseases and conditions for which such formulations are indicated.

BACKGROUND OF THE INVENTION

Liraglutide [Arg34, Lys26 (Ne-(?-Glu(Na-hexadecanoyl)))-GLP-1 (7-37)] an analog of human GLP-1 acts as a GLP-1 receptor agonist. It binds to GLP-1 receptors and increases intracellular cyclic AMP (cAMP) which leads to stimulation of insulin secretion in a glucose-dependent manner. Liraglutide also lowers blood triglyceride levels, decreases appetite and inhibits body weight gain. Liraglutide can be used clinically as an adjunct to diet and exercise to improve glycemic control, in chronic weight management and to reduce the risk of major adverse cardiovascular events in adults with type 2 diabetes mellitus and established cardiovascular disease. Liraglutide structurally represented as:

Therapeutic peptides are widely used in medical practise. Pharmaceutical formulations of such therapeutic peptides are required to have a good stability in order to be suitable for common use. However, peptide formulations are inherently unstable due to sensitivity towards chemical and physical degradation. Chemical degradation involves change of covalent bonds, such as oxidation, hydrolysis, racemization or crosslinking and deamindation. Physical degradation involves conformational changes relative to the native structure of the peptide, which may lead to oligomerization, aggregation, precipitation or adsorption to surfaces. These consequently lead to loss in biological activity of molecule.

In general, peptides encompassing 29-39 amino acids have a high degree of homology and they share a number of properties, notably their tendency to aggregate and formation of insoluble fibrils. This property seems to encompass a transition from a predominant alpha-helix conformation to beta-sheets. Aggregation of the glucagon-like peptides are mainly seen when solutions of the peptides are stirred or shaken, at the interface between solution and gas phase (air), and at contact with hydrophobic surfaces such as Teflon®. The inclusion of isotonicity agents in peptide containing pharmaceutical formulations is widely known and one of the more common isotonic agents used in such formulations is mannitol. It is known that mannitol causes problems during the production of peptide formulations as it crystallizes resulting in deposits in the production equipment or in the final product. Such deposits increase the need to clean the filling equipment during production of the formulation and this results in reduced production capability. In addition, such deposits may also result in reduced yield of the final product since vials/cartridges containing the peptide formulation may need to be discarded if particles are present. Further it was observed that in peptide formulations to be administered by injection, the presence of mannitol results in clogging of injection devices. Thus, various treatments and addition of excipients must often be applied to pharmaceutical formulations of the glucagon-like peptides including Liraglutide, in order to improve their stability. The in-use period where the product may be transported and shaken daily at ambient temperature preferably should be several weeks. Thus, there is a need for pharmaceutical formulations of Liraglutide which have improved stability.

Several attempts have been made in the prior art to provide optimal and stable injection formulations of Liraglutide. U.S. Patent No. 8,114,833 discloses a pharmaceutical formulation comprising a Liraglutide, a disodium phosphate dihydrate buffer, and propylene glycol, wherein said propylene glycol is present in said formulation in a final concentration of from about 1 mg/mL to about 100 mg/mL, and wherein said formulation has a pH of from about 7.0 to about 10.0. U.S. Patent No. 8,846,618 discloses a pharmaceutical formulation comprising a Liraglutide, an isotonic agent, a buffer, and a preservative, wherein said pharmaceutical formulation has a pH of from 7.5 to 9.4. U.S. Patent No. 8,614,181 discloses a method for increasing the shelf-life of a pharmaceutical formulation comprising a Liraglutide, a buffer and a preservative, by adjusting pH (8.5 to 11.5) of bulk peptide prior to freeze drying or formulation. U.S. Patent No. 8,114,959 discloses a method for increasing the shelf-life of a pharmaceutical formulation comprising a glucagon-like peptide, by the preparing a peptide solution having pH from 9.0 to 12.5 and drying it (prior to dissolving the peptide in buffer). U.S. Patent No. 8,748,376 discloses a method for preparation of a shelf-stable pharmaceutical formulation comprising a glucagon-like peptide, by preparing a peptide solution having pH from 8.0 to 10.5 and heating it, to form stable formulation. PCT Publication No. WO2016038521A1 discloses a pharmaceutical formulation comprising: (i) Liraglutide; (ii) a buffer selected from the group consisting of dipotassium phosphate, sodium bicarbonate, and disodium phosphate anhydrous; (iii) propylene glycol; and (iv) a preservative. PCT Publication No. WO2017147783A1 discloses a pharmaceutical formulation comprising Liraglutide, which is prepared by mixing Liraglutide and other excipients in a solvent until homogeneous, with a stirring speed of 500 ~ 1100rpm stirred, and adjusting the pH 7.5 to 9.5.

Currently Liraglutide is marketed under brand name VICTOZA® which is given by subcutaneous injection. It is indicated as an adjunct to diet and exercise in patients with type 2 diabetes, developed by Novo Nordisk. In the United States, it is being marketed under the brand name SAXENDA® as an adjunct to diet and exercise in patients with obesity. However, the currently available formulations of Liraglutide are associated with gastrointestinal side effects and the high clearance limits its usefulness. Further VICTOZA®, & SAXENDA® require storage at refrigerated temperatures prior to first use and should be thrown away after 30 days of the first use. Further the recommended dosing schedule for Liraglutide (Victoza) is to initiate therapy with the 0.6 mg dose daily for 1 week and then titrate to 1.2 mg; After 1 week of taking 1.2 mg, the dose can be increased to 1.8 mg if the 1.2 mg dose does not result in the desired glycemic control. Given the delicate stability and complex dosage regimen of Liraglutide, there remains a need to develop alternative therapeutically effective formulations of Liraglutide that are simple, easy to manufacture, and stable. Therefore, the inventors of the present invention have developed pharmaceutical formulations of Liraglutide using alternative buffers. Further, the formulations of the present invention are stable and easy to manufacture. The alternate buffers used in the present invention stabilizes the pH of the formulation between about 7.0 and 10.0 and alleviates the limitations in the art by providing better stable Liraglutide injectable preparations with reduced clogging of injection devices

SUMMARY OF INVENTION

Aspects of the present invention relates to stable pharmaceutical formulations comprising: (i) Liraglutide; (ii) a buffer selected from the group consisting of CAPS, CAPSO, CHES, AMP, AMPSO, AMPD, Ammonia, Methyl amine, Taurine, Tris, Boric acid, Monoethanolamine, Meglumine, Diethanolamine, Histidine, Aspartic acid, Glutamic acid, L- arginine, Glycine, Potassium chloride, Sodium Lactate, Sodium Borate, Calcium Lactate, and Calcium Carbonate; (iii) propylene glycol; and (iv) a preservative.

Aspects of the present invention relates to a multi-step process with tightly controlled by in-process controls for preparing the said Liraglutide formulations that exhibit reduced deposits on production equipment and in the final product.
Aspects of the present invention relates to method of safe and controlled delivery of said Liraglutide formulation with reduced clogging, from a pen injection device which is specifically adapted to provide for multiple measured injections. An another aspect of the present invention relates to the said stable Liraglutide formulation, wherein the formulation is present as a reservoir in a multiple dose pen injection device, and the device being adapted to subcutaneously inject the said reservoir in multiple doses, while the reservoir remains stable.

Aspects of the present invention relates to a method of using the said stable Liraglutide formulations as an adjunct to diet and exercise to improve glycemic control, in chronic weight management and to reduce the risk of major adverse cardiovascular events in adults with type 2 diabetes mellitus and established cardiovascular disease. The present invention further relates to methods of treatment using the said pharmaceutical formulations of the invention by administering an amount effective to combat the disease, condition, or disorder for which administration of Liraglutide in the formulation is indicated.

In an aspect of the present invention, the buffering agents stabilizes the pH of the Liraglutide formulation, wherein the preferred buffering agents are selected from CAPS (N-cyclohexyl-3-aminopropanesulfonic acid) buffer, Calcium Carbonate buffer, Calcium Lactate buffer, Tris buffer, CAPSO (N-cyclohexyl-2-hydroxyl-3-aminopropanesulfonic acid) buffer, Sodium Borate buffer, Sodium Lactate buffer, CHES (N-Cyclohexyl-2-aminoethanesulfonic acid) buffer, Histidine, Aspartic acid, Glutamic acid, L- arginine, Glycine, Potassium chloride buffer, Diethanolamine buffer, AMP (2-Amino-2-methyl-1-propanol) buffer, Ammonia buffer, Methyl amine buffer, AMPSO (N-(1,1-Dimethyl-2-hydroxyethyl)-3-amino-2-hydroxypropanesulfonic acid) buffer, Taurine buffer, Boric acid buffer, AMPD (aminomethyl propanediol) buffer, Monoethanolamine buffer, Meglumine buffer, buffer and mixtures thereof.

The formulations of the present invention remain stable for significant time and at different temperatures/conditions, during storage/transport and in-use period, after exposure to multiple freeze-thaw cycles, or after being subjected to different production process (including stirring, filtration and pH adjustment etc) which can cause aggregation or damage to their secondary structure.

DETAILED DESCRIPTION OF THE INVENTION

In embodiments, the pharmaceutical formulation provided herein comprises (i) Liraglutide; (ii) propylene glycol; (iii) preservative; and (iv) buffer selected from the group consisting of CAPS, CAPSO, CHES, AMP, AMPSO, AMPD, TAPS, Ammonia, Methyl amine, Taurine, Tris, Boric acid, Monoethanolamine, Meglumine, Diethanolamine, Histidine, Aspartic acid, Glutamic acid, L- arginine, Glycine, Potassium chloride, Sodium Lactate, Sodium Borate, Calcium Lactate, Calcium Carbonate and mixtures thereof.

In embodiments, the preservative is selected from the group consisting of phenol, m-cresol, thiomerosal, methyl paraben, propyl paraben, butyl paraben, chlorobutanol, 2-phenylethanol, benzyl alcohol and phenoxyethanol. In a preferred embodiment, the preservative is phenol.

In embodiments, the formulation of the present invention has a pH of between about 7.0 to about 11. Suitable pH-adjusting agents that can be used in the formulation of the present invention is selected from but not limited to a group comprising hydrochloric acid, N-methyl glucamine, sodium hydroxide, acetic acid, potassium hydroxide, phosphoric acid, lactic acid, and mixtures thereof. The pH of the solution is adjusted to about 7.0 to about 11.0. Preferably, the pH is adjusted to about 7.0 to about 10.0. More preferably the pH is adjusted to about one of the following values, i.e., 7.0, 7.2, 7.8, 7.9, 8.1, 8.2, 8.3, 8.4 or 9.8. The term "about" as used herein in relation to pH means ±0.1 pH units from the specified value.

In embodiments, Propylene glycol is preferably used as an isotonic agent in the pharmaceutical formulations of the present invention, where the propylene glycol is present in a concentration of 1-100 mg/ml. However, it can be replaced or used in combination with sodium chloride, potassium chloride, mannitol, sorbitol, trehalose, lactitol, xylitol, glycerol, sucrose, glycine, lactose, glucose, maltose, lysine, isoleucine, aspartic acid, L- glycine, L-histidine, arginine, myo-inositol, polyethylene glycol, or combinations thereof.

In embodiments, the pharmaceutical formulation provided herein further comprises a stabilizers and other excipients including a surfactant, and a chelating agent. Suitable examples of stabilizers include polyethylene glycol or its derivatives, polyvinyl alcohol, polyvinylpyrrolidone, carboxymethyl cellulose, sodium chloride, L- glycine, L-histidine, imidazole, arginine, lysine, isoleucine, aspartic acid, tryptophan, threonine, or mixtures thereof. Suitable examples of surfactants include polysorbate, poloxamers, ethylene/polypropylene block polymers, lecithins, alcohols, sodium lauryl sulfate, bile acids and salts thereof, polymeric surfactants, long-chain fatty acids, phospholipids, ethoxylated castor oil, polyglycolyzed glycerides, acetylated monoglycerides, sorbitan fatty acid esters, polyoxyethylene sorbitan fatty acid esters, monoglycerides, diglycerides, glycerol, glycerophospholipids, glyceroglycolipids, sphingophospholipids, sphingoglycolipids, docusate sodium, docusate calcium, docusate potassium, and mixtures thereof. Suitable examples of chelating agents include ethylenediaminetetraacetic acid or its salts, and mixtures thereof.

In one of the embodiments, the stable pharmaceutical formulation provided herein comprises (i) Liraglutide, (ii) Tromethamine as buffering agent, (iii) propylene glycol and (iv) phenol, wherein the formulation has a pH of 8.0 to 8.3.
In one of the embodiments, the stable pharmaceutical formulation provided herein comprises (i) Liraglutide (ii) Aspartic acid as buffering agent (iii) propylene glycol and (iv) phenol, wherein the formulation has a pH of 8.0 to 8.3.

In one of the embodiments, the stable pharmaceutical formulation provided herein comprises (i) Liraglutide, (ii) Glutamic acid as buffering agent (iii) propylene glycol and (iv) phenol, wherein the formulation has a pH of 8.0 to 8.3.

In one of the embodiments, the stable pharmaceutical formulation provided herein comprises (i) Liraglutide, (ii) L- arginine as buffering agent (iii) propylene glycol and (iv) phenol, wherein the formulation has a pH of 8.0 to 8.3.

In one of the embodiments, the stable pharmaceutical formulation provided herein comprises (i) Liraglutide (ii) Glycine as buffering agent (iii) propylene glycol and (iv) phenol, wherein the formulation has a pH of 8.0 to 8.3.

In one of the embodiments, the stable pharmaceutical formulation provided herein comprises (i) Liraglutide (ii) Histidine as buffering agent (iii) propylene glycol and (iv) phenol, wherein the formulation has a pH of 8.0 to 8.3.

In one of the embodiments, the stable pharmaceutical formulation provided herein comprises (i) Liraglutide (ii) Meglumine as buffering agent (iii) propylene glycol and (iv) phenol, wherein the formulation has a pH of 8.0 to 8.3.

In embodiments, the pharmaceutical formulation provided herein can be prepared by multi step process which is tightly controlled by in-process controls. In one embodiment, the process comprises the following steps:
a. Prepare a first solution by dissolving buffer in aqueous medium;
b. Add and dissolve propylene glycol to the solution prepared in step a);
c. Add and dissolve preservative to the solution prepared in step b);
d. Add and dissolve Liraglutide to the solution prepared in step c);
e. Check and adjust the pH of the solution prepared in step d) and it make it up to 100% of the batch size with aqueous medium; and
f. Filter the solution prepared in step e) to fill the filtrated solution into cartridges.

In another embodiment, the process comprises the following steps:
a. Mix all the ingredients (Except Liraglutide) in aqueous medium and the pH was adjusted to 7.0 with sodium hydroxide/Hydrochloric acid.
b. Add and dissolve Liraglutide in the solution prepared in step a).
c. Adjust the pH of above the solution prepared in step b) to final value using pH adjusting agent (Sodium hydroxide/ Hydrochloric acid).
d. The resultant bulk solution was filtered using 0.2 micron filter and filled in cartridges.

In another embodiment, the method for preparing a peptide formulation comprises:
a. Prepare a first solution by dissolving preservative and buffer in water;
b. Add propylene glycol to the first solution;
c. Mix the first solution with a second solution containing Liraglutide dissolved in water; and
d. Adjust the pH of the mixture in c) to the desired pH.
e. The resultant bulk solution was filled in cartridges

In another embodiment, the method for preparing a peptide formulation comprises:
a. Collect the water for injection required for batch size in compounding vessel;
b. Cool the water for injection to 2-8°C with nitrogen purging and chill water circulation through jacketed vessel;
c. Collect 80% of WFI required for batch size in compounding vessel and transfer remaining WFI to make up vessel;
d. To the above step, add Propylene Glycol under continuous to get clear colourless solution.
e. To the above step, add Phenol under continuous to get clear colourless solution;
f. To the above step, add a buffering agent selected from Tromethamine, Aspartic acid, Glutamic acid, L- arginine, Glycine, Histidine and Meglumine under continuous to get clear colourless solution;
g. To the above step, add Liraglutide API under continuous to get clear colourless solution;
h. Check the pH of the solution and adjust the pH of the solution with HCl or Sodium Hydroxide to 8.00 - 8.30;
i. Finally make up the volume to 100% of batch size.

In embodiments, the pharmaceutical formulation provided herein is prepared by a multi-step process that is tightly controlled by in-process controls, the suitable process controls that can be employed in the multistep processes of the present invention are selected from the following parameters but not limited to a) stirring at speed of 100 to 1400 rpm, b) stirring until homogeneous mixture is formed, c) filtration with 0.2µm membrane at a pressure of 0.05 to 0.18mpa and combinations thereof. In one embodiment, the stabilized formulation process may optionally comprise steps including but not limited to a) heating, b) drying, c) protection from light i.e., formulation under sodium vapor lamp d) Nitrogen purging and pH adjustment.

In one embodiment, the stabilized formulation of the present invention can be sterilized. Non-limiting examples of sterilization techniques include filtration through a bacterial-retaining filter, terminal sterilization, and incorporation of sterilizing agents, irradiation, and heating.

The term "buffer," as used herein, refers to a chemical compound added to a Liraglutide formulation to prevent the pH from changing with time. Certain non-limiting examples of buffering agents include CAPS, CAPSO, CHES, AMP, AMPSO, AMPD, TAPS, Ammonia, Methyl amine, Aspartic acid, Glutamic acid, L- arginine or its salt, Glycine or its salt, Histidine, glycylglycine, Taurine, Boric acid, Monoethanolamine, Meglumine, Diethanolamine, Potassium chloride, Sodium Lactate, Sodium Borate, Calcium Lactate, and Calcium Carbonate, Sodium Acetate, Sodium phosphate, Dipotassium phosphate, Sodium bicarbonate, & Disodium phosphate anhydrous, Sod ium Bicarbonate, Arginin, Sodium Citrate, Lysine, Sodium Carbonate, Disodium hydrogen phosphate, Dibasic Potassium phosphate, Sodium Citrate Dihydrate, Sodium dihydrogen phosphate, Citrate, HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid)), disodium phosphate dehydrate, Bicine (N,N-Bis(2-hydroxyethyl)glycine), Glycine, disodium phosphate dihydrate or their salts or combinations thereof. More preferably Diethanolamine buffer, Tris Buffer, Monoethanolamine Buffer, Meglumine Buffer, Aspartic acid, Glutamic acid, L- arginine, Glycine, Histidine, Ammonia Buffer and mixtures thereof.

Within the scope of the description of the invention, Liraglutide and any of its addition salts are referred to as "Liraglutide". Also, unless otherwise stated, the terms "active ingredient" and "active substance" refer to Liraglutide and any of its pharmaceutically salts. The effective therapeutic amount of Liraglutide is advantageously about 0.4mg/day to 3 mg /day. When the active principle is a Liraglutide pharmaceutically acceptable salt, the salt content in the formulation is such that the latter comprises from 6 mg/ml of Liraglutide base.

In embodiments, the expression "stable pharmaceutical formulation" is intended to mean stable during different temperatures for a long period of time, shelf stable formulation, stable in the device. As used herein, the term "Stabilized formulation” further refers to a stable solution for therapeutic use, containing Liraglutide.

The term ‘injection device’ as used herein means an injection assembly which can house a reservoir of a multiple sterile doses of Liraglutide in solution. Preferably the reservoir is in a cartridge. Further the multiple dose injection device includes a pen with dose dialing facility that allows the user to select and inject a desired dose of Liraglutide formulation. The device includes an injection assembly in which the reservoir of Liraglutide is in a cartridge which is in-built or replaceable in the multiple dose pen injection device. The device does not involve the withdrawal of the drug solution from a separate storage container as is the case in conventional syringe-vial assembly. That is, the sterile solution of Liraglutide can be directly administered from the pen device, such that there is no requirement of withdrawal of the Liraglutide solution from a separate storage container as is the case in a conventional syringe-vial assembly.
The following examples are intended to serve as illustrations of the present invention and not intended to be limiting in any manner.

EXAMPLES
Example 1:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Diethanolamine Quantity sufficient (0.01 to 25 mg)
3 Propylene glycol 14 mg
4 Phenol 5.5 mg
5 Water for Injection q.s to 1 mL

Example 2:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Tromethamine (Tris) Quantity sufficient (0.01 to 30 mg)
3 Propylene glycol 14 mg
4 Phenol 5.5 mg
5 Water for Injection q.s to 1 mL

Example 3:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Monoethanolamine Quantity sufficient (0.01 to 10 mg)
3 Propylene glycol 14 mg
4 Phenol 5.5 mg
5 Water for Injection q.s to 1 mL

Example 4:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Meglumine Quantity sufficient (0.01to 10 mg)
3 Propylene glycol 14 mg
4 Phenol 5.5 mg
5 Water for Injection q.s to 1 mL

Example 5:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Ammonia Quantity sufficient (0.01 to 10 mg)
3 Propylene glycol 14 mg
4 Phenol 5.5 mg
5 Water for Injection q.s to 1 mL

Example 6:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Calcium Carbonate 0.05 mg to 100 mg (pH 9.0)
3 Propylene glycol 4.66 mg
4 Phenol 1.83 mg
5 Water for Injection q.s to 1 mL

Example 7:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Calcium Lactate 0.05 mg to 80 mg (pH 6.0–8.5)
3 Propylene glycol 4.66 mg
4 Phenol 1.83 mg
5 Water for Injection q.s to 1 mL

Example 8:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Sodium Borate 0.05 mg to 60 mg (pH 9.0–9.6)
3 Propylene glycol 4.66 mg
4 Phenol 1.83 mg
5 Water for Injection q.s to 1 mL

Example 9:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Sodium Lactate 0.01 to 48 mg (pH 7)
3 Propylene glycol 4.66 mg
4 Phenol 1.83 mg
5 Water for Injection q.s to 1 mL

Example 10:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Boric acid 0.01 to 10 mg
3 Propylene glycol 4.66 mg
4 Phenol 1.83 mg
5 Water for Injection q.s to 1 mL

Example 11:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Taurine 0.01 to 10 mg
3 Propylene glycol 4.66 mg
4 Phenol 1.83 mg
5 Water for Injection q.s to 1 mL

Example 12:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Methyl amine 0.01 to 10 mg
3 Propylene glycol 4.66 mg
4 Phenol 1.83 mg
5 Water for Injection q.s to 1 mL

Example 13:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 AMPD 0.01 to 50 mg
3 Propylene glycol 4.66 mg
4 Phenol 1.83 mg
5 Water for Injection q.s to 1 mL

Example 14:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 AMPSO 0.01 to 50 mg
3 Propylene glycol 4.66 mg
4 Phenol 1.83 mg
5 Water for Injection q.s to 1 mL

Example 15:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 AMP 0.01 to 50 mg
3 Propylene glycol 4.66 mg
4 Phenol 1.83 mg
5 Water for Injection q.s to 1 mL

Example 16:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 CHES 0.01 to 50 mg
3 Propylene glycol 4.66 mg
4 Phenol 1.83 mg
5 Water for Injection q.s to 1 mL

Example 17:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 CAPSO 0.01 to 50 mg
3 Propylene glycol 4.66 mg
4 Phenol 1.83 mg
5 Water for Injection q.s to 1 mL

Example 18:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 CAPS 0.01 to 50 mg
3 Propylene glycol 4.66 mg
4 Phenol 1.83 mg
5 Water for Injection q.s to 1 mL

Example 19:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Histidine 0.1mg to 15mg
3 Propylene glycol 14 mg
4 Phenol 5.5 mg
5 Sodium Hydroxide QS to pH
6 Hydrochloric acid QS to pH
7 Water for Injection q.s to 1 mL

Example 20:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Aspartic acid 0.1mg to 20mg
3 Propylene glycol 14 mg
4 Phenol 5.5 mg
5 Sodium Hydroxide QS to pH
6 Hydrochloric acid QS to pH
7 Water for Injection q.s to 1 mL

Example 21:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Glutamic acid 0.1mg to 20mg
3 Propylene glycol 14 mg
4 Phenol 5.5 mg
5 Sodium Hydroxide QS to pH
6 Hydrochloric acid QS to pH
7 Water for Injection q.s to 1 mL

Example 22:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 L-arginine 0.1mg to 30mg
3 Propylene glycol 14 mg
4 Phenol 5.5 mg
5 Sodium Hydroxide QS to pH
6 Hydrochloric acid QS to pH
7 Water for Injection q.s to 1 mL

Example 23:
S.No Ingredients Qty/mL
1 Liraglutide (free-base, anhydrous) 6 mg
2 Glycine 0.1mg to 30mg
3 Propylene glycol 14 mg
4 Phenol 5.5 mg
5 Sodium Hydroxide QS to pH
6 Hydrochloric acid QS to pH
7 Water for Injection q.s to 1 mL

Stability can be measured in many ways, including measuring osmolarity or by visually inspecting the formulations in daylight with a dark background for any signs of turbidity, changes in color or clarity, or any other visible precipitates.

Example No. Buffering agent used Description pH Osmolarity
Example-1 Diethanolamine buffer Clear, colorless solution 8.35
265 mOsm/Kg
Example-2 Tris buffer Clear, colorless solution 7.88 272 mOsm/Kg
Example-3 Monoethanolamine buffer Clear, colorless solution 8.12 252 mOsm/Kg
Example-4 Meglumine buffer Clear, colorless solution 8.40
256 mOsm/Kg
Example-5 Ammonia Buffer Clear, colorless solution 7.89
245 mOsm/Kg

Based on the study, it can be concluded that the formulation maintains the stability of the drug product. ,CLAIMS:WE CLAIM:

1. A stable pharmaceutical formulation comprising Liraglutide, a buffering agent, propylene glycol and a preservative, wherein the formulation has a pH of 7.0 to 10.0.

2. The formulation as claimed in claim 1, wherein the buffering agent of the formulation can be selected from CAPS (N-cyclohexyl-3-aminopropanesulfonic acid) buffer, Calcium Carbonate buffer, Calcium Lactate buffer, Tromethamine (Tris) buffer, CAPSO (N-cyclohexyl-2-hydroxyl-3-aminopropanesulfonic acid) buffer, Sodium Borate buffer, Sodium Lactate buffer, CHES (N-Cyclohexyl-2-aminoethanesulfonic acid) buffer, Potassium chloride buffer, Diethanolamine buffer, AMP (2-Amino-2-methyl-1-propanol) buffer, Ammonia buffer, Methyl amine buffer, AMPSO (N-(1,1-Dimethyl-2-hydroxyethyl)-3-amino-2-hydroxypropanesulfonic acid) buffer, Aspartic acid, Glutamic acid, L- arginine, Glycine, Histidine, Taurine buffer, Boric acid buffer, AMPD (aminomethyl propanediol) buffer, Monoethanolamine buffer, Meglumine buffer, and mixtures thereof.

3. The formulation as claimed in claim 1, wherein the preservative is selected from the group consisting of phenol, m-cresol, thiomerosal, methyl paraben, propyl paraben, butyl paraben, chlorobutanol, 2-phenylethanol, benzyl alcohol and phenoxyethanol.

4. The formulation as claimed in claim 1, further comprises of pharmaceutical excipients such as pH adjusting agents, water, stabilizing agents or mixtures thereof.

5. The formulation as claimed in claim 1, wherein the formulation is present as a reservoir in a multiple dose pen injection device and the device being adapted to subcutaneously inject the said reservoir in multiple doses with reduced clogging, while the reservoir remains stable.

6. A stable pharmaceutical formulation comprising (i) Liraglutide, (ii) a buffering agent selected from Tromethamine, Aspartic acid, Glutamic acid, L- arginine, Glycine, Histidine and Meglumine, (iii) propylene glycol and (iv) phenol, wherein the formulation has a pH of 8.0 to 8.3.

7. A multi-step process with tight in-process controls for preparing stable Liraglutide formulations that exhibit reduced deposits on production equipment and in the injection device, wherein the formulation is prepared by mixing Liraglutide with excipients using a solvent.

8. The process of claim 7, wherein the multi-step process for preparing the said Liraglutide formulation comprises the following steps:
a) Add propylene glycol to cool water for injection
b) Add preservative to the solution of step a) and stir
c) Add buffering agent to the solution of step b) and stir
d) Add Liraglutide to the solution of step c) and stir
e) Adjust the pH of step d) solution to 7.0 - 10.0 by using a suitable pH adjusting agent
f) Make up the volume to required batch size and filter the solution through filter.

9. The process of claim 7, wherein the multi-step process for preparing of the said Liraglutide formulation comprises the following steps:
a) Add propylene glycol to cool water for injection
b) Add phenol to the solution of step a) and stir
c) Add buffering agent selected from Tromethamine, Aspartic acid, Glutamic acid, L- arginine, Glycine, Histidine and Meglumine, to the solution of step b) and stir
d) Add Liraglutide to the solution of step c) and stir
e) Adjust the pH of step d) solution to about 8.0 – about 8.3 by using HCl or Sodium Hydroxide
f) Make up the volume to required batch size and filter the solution through filter.

10. A method of using the said stable Liraglutide formulations as an adjunct to diet and exercise to improve glycemic control, in chronic weight management and to reduce the risk of major adverse cardiovascular events in adults with type 2 diabetes mellitus and established cardiovascular disease.