Anti-il17 antibody formulations

Abstract

: The present disclosure relates to the field of pharmaceutical products, and in particular relates to anti-IL17 antibody formulations and the processes of making 5 such pharmaceutical products, more particularly, the present disclosure relates to antibody formulation comprising a high concentration of anti-IL17 antibody and pharmaceutically acceptable excipients selected from the group comprising of buffer(s), stabilizer(s), viscosity modifier(s), antioxidants and surfactant(s).

Description

[0001] ANTI-IL17 ANTIBODY FORMULATIONS

[0002] Field of Invention:

[0003] The present disclosure relates to the field of antibody formulations, in particular relates to antiInterleukin- 17 antibody formulations and the process for preparation thereof.

[0004] Background of Invention:

[0005] Long-term storage of antibodies and proteins has been linked to challenges in stability, particularly at ambient temperatures. The antibody and / or proteins often face issues from various physical and chemical reactions like aggregation, deamidation, oxidation, clipping, isomerization, denaturation, leading to an increase in degradation product levels and / or loss of biological activity. Furthermore, high concentration formulations of antibody or protein (typically containing 25 to 300 mg / ml of active molecule) often lead to increased solution viscosities, further leading to stability, manufacturing and delivery challenges. To mitigate these issues, it is essential to develop effective formulation strategies that enhances the stability and preserve integrity of the biologies during storage.

[0006] There are few anti-Interleukin-17 (anti-IL-17) antibodies approved for therapeutic use and several are under exploration for the therapeutic use. At least three anti-IL-17 antibodies viz. Secukinumab, Ixekizumab, and Bimekizumab are approved for therapeutic indications such as treatment of dermatological and autoimmune disorders like rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis, chronic plaque-type psoriasis, and hidradenitis suppurativa.

[0007] Secukinumab is first disclosed by Novartis in W02006013107. It is a recombinant human monoclonal IgGl / K antibody, has molecular mass of approximately 151 kDa, and is expressed in a recombinant Chinese hamster ovary (CHO) cell line. The said antibody encompasses a free cysteine at 97thposition on light chain which tends to oxidize and thereby leads to loss of stability and biological activity.

[0008] In past, efforts have been made to stabilize secukinumab using various pharmaceutically acceptable excipient. International patent application WO2012059598 discloses secukinumab formulation comprising of histidine buffer, surfactant, and sucrose. Another international patent application W02016103153 discloses a liquid formulation of secukinumab comprising of a stabilizer i.e. methionine, wherein said formulation is stored in a container having a headspace comprising less than about 12% oxygen. A Chinese patent application, CN118056572A disclosesa liquid secukinumab-hyaluronidase formulation comprising of excipients methionine and trehalose as stabilizers. Further, few more international patent applications, viz. W02020233540, WO2024196220 & WO2022135395 disclose liquid secukinumab formulations comprising of methionine and / or sugar alcohol as stabilizer, while WO2024156841 and W02022113105A1 use various buffers to stabilize the liquid formulations of secukinumab. Still, there is a need in the art for stable and delivery friendly secukinumab formulation(s).

[0009] Summary of invention:

[0010] The present disclosure relates to a stable antibody formulation comprising anti-IL17 antibody and one or more stabilizers selected from the group comprising of pentetic acid, pyridoxine, proline, phenylalanine, and pharmaceutically acceptable salts thereof.

[0011] In one embodiment, the present disclosure relates to a stable anti-IL17 antibody formulation comprising one or more buffers, surfactant and two stabilizers, wherein the first stabilizer is selected from the group comprising of sugars and sugar alcohols; and the second stabilizer is selected from the group comprising of pentetic acid, pyridoxine, proline, phenylalanine, and pharmaceutically acceptable salts thereof.

[0012] In another embodiment, the present disclosure relates to a stable secukinumab formulation comprising one or more buffers, surfactant and two stabilizers, wherein the first stabilizer is selected from the group comprising of sugars and sugar alcohols; and the second stabilizer is selected from the group comprising of pentetic acid, pyridoxine, proline, phenylalanine, and pharmaceutically acceptable salts thereof.

[0013] In yet another embodiment, the present disclosure relates to a stable high concentration formulation of secukinumab comprising one or more buffers, surfactant and at least two stabilizers, wherein the first stabilizer is selected from the group comprising of sugars and sugar alcohols comprising of sucrose, trehalose, raffinose, maltose, sorbitol, xylitol, mannitol; and the second stabilizer is selected from the group comprising of pentetic acid, pyridoxine, proline, phenylalanine, and pharmaceutically acceptable salts thereof; wherein the formulation is devoid of methionine.

[0014] Definitions:

[0015] The terms "comprises," "comprising," “including,” and “having,” are open ended transitional phrases and therefore specify the presence of stated features, integers, steps, operations, elements, modules, units and / or components, but do not forbid the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof.The term as used in the present disclosure, the forms "a,” "an," and "the" may be intended to include the plural forms as well, unless the context clearly suggests otherwise.

[0016] The term “buffer” (or “buffer system”) is well known in the art. Herein, the terms “buffer” and “buffer system” may be used interchangeably. Abuffered solution comprises buffering species (i.e. the species in dynamic equilibrium with one another, e.g. acetate / acetic acid for an acetate buffer system or histidine / histidine hydrochloride for a histidine buffer system) that contribute to the buffer / buffer system, and thus comprises a mixture of an acid (usually a weak acid, e.g. acetic acid or histidine) and its conjugate base (e.g. sodium acetate or histidine hydrochloride).

[0017] The term “surfactant” is well known in the art. Herein, the term surfactant refers to a chemical compound that decrease the surface tension or interfacial tension between two liquids, a liquid and a gas, or a liquid and a solid.

[0018] The term “stabilizer” (or stabilizers) is well known in the art. Herein the term stabilizer refers to a compound that provides stability to the antibody or protein molecule by reducing the oxidation and / or aggregation of antibody molecules in a formulation. The stabilizer may have additional properties like antioxidant, viscosity modifier and / or chelating agent.

[0019] The term “pharmaceutically acceptable salt” is well known in the art. Herein the term “pharmaceutically acceptable salt” refers to a compound prepared from an acid which form nontoxic acid anions such as the hydrochloride, hydrobromide, sulphate, phosphate or acid phosphate, acetate, maleate, fumarate, lactate, tartrate, citrate and gluconate salts.

[0020] Figures:

[0021] Fig. 1 (a) is a size exclusion - HPLC chromatogram studied on Day 0 representing the amount of high molecular weight species, low molecular weight species and the monomeric concentration of secukinumab present in a formulation.

[0022] Fig. 1 (b) is a size exclusion - HPLC chromatogram of samples stored at 40 °C for 2 weeks, representing the amount of high molecular weight species, low molecular weight species and the monomeric concentration of secukinumab present in a formulation.

[0023] Fig. 1 (c) is a size exclusion - HPLC chromatogram studied of samples stored at 2-8 °C for 3 months, representing the amount of high molecular weight species, low molecular weight species and the monomeric concentration of secukinumab present in a formulation.Fig. 2 (a) is an Ion Exchange - High-Performance Liquid (IEX-HPLC) Chromatogram on Day 0 representing the concentration of acidic, basic and main species of secukinumab present in a formulation.

[0024] Fig. 2 (b) is an Ion Exchange - High-Performance Liquid (IEX-HPLC) chromatogram of samples stored at 40 °C for 2 weeks, representing the concentration of acidic, basic and main species of secukinumab present in a formulation.

[0025] Fig. 2 (c) is an Ion Exchange - High-Performance Liquid (IEX-HPLC) chromatogram studied of samples stored at 2-8 °C for 3 months, representing the concentration of acidic, basic and main species of secukinumab present in a formulation.

[0026] Description:

[0027] The present disclosure provides an antibody formulation and a process for preparing the said formulation. It is understood that each feature or embodiment, or combination, described herein is a non-limiting, illustrative example of any of the embodiments of the disclosure and, as such, is meant to be interchangeable and combinable with any other embodiments and / or features as disclosed herein.

[0028] In one of the embodiments, the present disclosure relates to anti-IL-17 antibody formulation comprising a high concentration antibody and one or more buffer(s), surfactant(s), and one or more stabilizers; wherein the stabilizer is selected from the group comprising of sucrose, trehalose, raffinose, maltose, sorbitol, xylitol, mannitol, pentetic acid , pyridoxine or its salts, proline or its salts, phenylalanine or its salts, and mixtures thereof.

[0029] In one of the embodiments, anti-IL-17 antibody is secukinumab.

[0030] In another embodiment of the present disclosure, the high concentration antibody formulation comprises of a secukinumab at a concentration of about 25 mg / ml to 300 mg / ml and one or more buffer(s), surfactant(s) and one or more stabilizers, wherein the stabilizer is selected from the group comprising of sucrose, trehalose, raffinose, maltose, sorbitol, xylitol, mannitol, pentetic acid, pyridoxine or its salts, proline or its salts, phenylalanine or its salts, and mixtures thereof.

[0031] In one more embodiment of the present disclosure, formulation according to present invention comprises of secukinumab and a combination of one or more buffers selected from histidine buffer, citrate buffer, acetate buffer, phosphate buffer, tris buffer and mixtures thereof; one or more surfactant selected from non-ionic surfactants, ionic surfactants, zwitterionic surfactants andmixtures thereof; and one or more stabilizers selected from the sucrose, trehalose, raffinose, maltose, sorbitol, xylitol, mannitol, pentetic acid, pyridoxine, proline, phenylalanine, and combinations thereof.

[0032] In one more embodiment of the disclosure, formulation according to present invention comprises of secukinumab and a combination of one or more buffers selected from histidine buffer, citrate buffer, acetate buffer, phosphate buffer, tris buffer and mixtures thereof; one or more surfactant selected from non-ionic surfactants, ionic surfactants, zwitterionic surfactants and mixtures thereof; and two stabilizers. One of the two stabilizers is selected from the sucrose, trehalose, raffinose, maltose, sorbitol, xylitol, mannitol; and another stabilizer is selected from the group comprising of pentetic acid, pyridoxine, proline, phenylalanine, and combinations thereof.

[0033] In one embodiment of the present disclosure, formulation according to present invention comprises of secukinumab and a combination of one or more excipients. In some embodiments pH of the formulation is in the range of about 5.0 to about 7.0; preferably pH of the formulation is in the range of 5.5 to 6.5, and more preferably pH is between 5.8 and 6.0. In some embodiments the pH of the formulation is about 5, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, or about 7. In some embodiments the pH of the formulation is 5 ± 0.5, 5.1± 0.5, 5.2± 0.5, 5.3± 0.5, 5.4± 0.5, 5.5± 0.5, 5.6± 0.5, 5.7± 0.5, 5.8± 0.5, 5.9± 0.5, 6± 0.5, 6.1± 0.5, 6.2± 0.5, 6.3± 0.5, 6.4± 0.5, 6.5± 0.5, 6.6± 0.5, 6.7± 0.5, 6.8± 0.5, 6.9± 0.5, or 7± 0.5. In one more embodiment of the present disclosure, the formulation comprises of pharmaceutically acceptable excipient(s), wherein the pharmaceutically acceptable excipient is one or more buffer(s) having optimal buffering capacity between pH of 5 and 7. In preferred embodiments, one or more buffer is selected from the group comprising of histidine buffer, citrate buffer, acetate buffer, phosphate buffer, tris buffer, and combinations thereof, at a concentration of 1 mM to 50 mM and a pH of 5.8 ±0.5. In some embodiments, the concentration of buffer is between 1 mM to 10 mM, 10 mM to 20 mM, 20 mM - 30 mM, 30 mM - 40 mM, or 40 mM to 50 mM. In some embodiments, the concentration of buffer is 1 mM ± 10%, 10 mM ± 10%, 15 mM ± 10%, 20 mM ± 10%, 25 mM ± 10%, 30 mM ± 10%, 35 mM ± 10%, 40 mM ± 10%, 45 mM ± 10% or 50 mM ± 10%. In another aspect of the embodiment, the formulation comprises of pharmaceutically acceptable excipient(s), wherein the pharmaceutically acceptable excipients are devoid of a buffer (buffer-free formulation).In another embodiment, the formulation comprises of pharmaceutically acceptable excipients, wherein the pharmaceutically acceptable excipient is one or more surfactant at a concentration of about 0.01% to about 0.4%. In preferred embodiments, one or more surfactant is selected from the group comprising of non-ionic surfactants, ionic surfactants, zwitterionic surfactants and combinations thereof. In a more preferred embodiment, the surfactant is a non-ionic surfactant and is selected from the group comprising of pol oxamer (e.g., pol oxamer 188) or polyoxyethylene sorbitan fatty acid esters (e.g. polysorbate 20, polysorbate 40, polysorbate 60 and / or polysorbate 80); preferably the surfactant is polysorbate 20. In one of the embodiments concentration of poloxamer or polyoxyethylene sorbitan fatty acid esters is about 0.01% to about 0.04% (w / v), about 0.04% to about 0.08% (w / v), about 0.08% to about 0.12% (w / v), about 0.12% to about 0.2% (w / v), about 0.01% to about 0.4% (w / v), about 0.2% to about 0.4% (w / v). about 0.01%, about 0.02%, about 0.04%, about 0.06%, about 0.08%, about 0.1%, about 0.2%, about 0.3%, about 0.4%,). In one embodiment, the formulation comprises about 0.01 to about 0.03% (w / v) polysorbate-20, more particularly 0.02% (w / v) polysorbate-20.

[0034] In one more embodiment of the present disclosure, the formulation comprises of pharmaceutically acceptable excipients, wherein the pharmaceutically acceptable excipient is one or more stabilizers selected from sugar, sugar alcohols, amino acids, salts, antioxidants, chelating agents, and combinations thereof, wherein (a) sugar and / or sugar alcohol stabilizers are selected from the group comprising of mono-, di- and tri-saccharides e.g. sucrose, trehalose, raffinose, maltose, sorbitol, xylitol, mannitol and combinations thereof; (b) amino acid stabilizers are selected from the group comprising of proline or its salts, phenylalanine or its salts, and combinations thereof; and (c) antioxidant or chelating agent stabilizers are selected from the group comprising of pentetic acid, pyridoxine or its salts.

[0035] In one embodiments, the concentration of first stabilizer (one or more selected from the group comprising of sucrose, trehalose, raffinose, maltose, sorbitol, xylitol, mannitol and combinations thereof) in formulation is in the range of about 10 mM to about 50 mM, about 50 mM too about 100 mM, about 100 mM to about 150 mM, about 150 mM to about 200 mM, about 200 mM to about 250 mM, about 250 mM to about 300 mM, about 100 mM, about 110 mM, about 120 mM, about 130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about 180 mM, about 190 mM, about 200 mM, about 210 mM, about 220 mM, about 230 mM, about 240 mM, about 250 mM, about 260 mM, about 270 mM, about 280 mM, about 290 mM, or about 300 mM. In one embodiment, the second stabilizer in formulation is pentetic acid at a concentration of about 1 pM to about 100 pM, about 1 pM to about 10 pM, about 10 pM to about 20 pM, about 20 pM to about 50 pM, about 50 pM to about 100 pM, about 1 pM± 10%, about 2 pM± 10%, about 3pM± 10%, about 4 pM± 10%, about 5 pM± 10%, about 6 pM± 10%, about 7 pM± 10%, about 8 pM± 10%, about 9 pM± 10%, about 10 pM± 10%, about 15 pM± 10%, about 20 pM± 10%. In another embodiment the second stabilizer in formulation is phenylalanine at a concentration of about 10 mM to about 100 mM, about 10 mM ± 10%, about 20 mM ± 10%, about 25 mM ± 10%, about 30 mM ± 10%, about 35 mM ± 10%, about 40 mM ± 10%, about 45 mM ± 10%, about 50 mM ± 10%, about 60 mM ± 10%, about 70 mM ± 10%, about 80 mM ± 10%, about 90 mM ± 10%, about 100 mM ± 10%.

[0036] In yet another embodiment the second stabilizer in formulation is proline at a concentration of about 10 mM to about 100 mM, about 10 mM ± 10%, about 20 mM ± 10%, about 25 mM ± 10%, about 30 mM ± 10%, about 35 mM ± 10%, about 40 mM ± 10%, about 45 mM ± 10%, about 50 mM ± 10%, about 60 mM ± 10%, about 70 mM ± 10%, about 80 mM ± 10%, about 90 mM ± 10%, about 100 mM ± 10%.

[0037] In yet another embodiment the second stabilizer in formulation is pyridoxine or its pharmaceutically acceptable salts at a concentration of about 10 mM to about 100 mM, about 10 mM ± 10%, about 20 mM ± 10%, about 25 mM ± 10%, about 30 mM ± 10%, about 35 mM ± 10%, about 40 mM ± 10%, about 45 mM ± 10%, about 50 mM ± 10%, about 60 mM ± 10%, about 70 mM ± 10%, about 80 mM ± 10%, about 90 mM ± 10%, about 100 mM ± 10%.

[0038] In another embodiment of the present disclosure, the formulation comprises of a buffer at a concentration of about 1 mM to about 50 mM; surfactant at a concentration of about 0.01% to about 0.4%; first stabilizer (stabilizer- 1) is one or more selected from the group comprising of sucrose, trehalose, raffinose, maltose, sorbitol, xylitol, mannitol and combinations thereof, wherein concentration of first stabilizer is about 10 mM to about 300 mM; and second stabilizer (stabilizer-2) is one or more selected from the group comprising of pentetic acid, pyridoxine, proline, phenylalanine, and combinations thereof, wherein concentration of second stabilizer is about 1 pM to about 100 mM. In one embodiment, the second stabilizer is pentetic acid at a concentration of about 1 pM to about 100 pM. In another embodiment the second stabilizer is phenylalanine at a concentration of about 10 mM to about 100 mM. In yet another embodiment the second stabilizer is proline at a concentration of about 10 mM to about 100 mM. In yet another embodiment the second stabilizer is pyridoxine or its pharmaceutically acceptable salts at a concentration of about 10 mM to about 100 mM.

[0039] In yet another embodiment of the present disclosure, the formulation comprises of (i) secukinumab at a concentration of about 25 mg / ml to about 300 mg / ml, more preferably 25 mg / ml or 150 mg / ml; (ii) one or more buffer at a concentration of about 1 mM to about 50 mM, wherein the buffer isselected from the group comprising of histidine buffer, citrate buffer, acetate buffer, phosphate buffer, tris buffer and combinations thereof; (iii) one or more surfactant at a concentration of about 0.01% to about 0.4%, wherein the surfactant is selected from the group comprising of non-ionic surfactants, ionic surfactants, zwitterionic surfactants and combinations thereof; (iv) one or more stabilizer-1 at a concentration of about 10 mM to about 300 mM, wherein stabilizer-1 is selected from the group comprising of sucrose, trehalose, raffinose, maltose, sorbitol, xylitol, mannitol and combinations thereof; (v) one or more stabilizer-2 at a concentration of about IpM to about 100 mM, wherein stabilizer-2 is selected from the group comprising of pentetic acid, pyridoxine, proline, phenylalanine, and / or pharmaceutically acceptable salts thereof.

[0040] In yet another embodiment of the present disclosure, the secukinumab formulation comprises buffer at a concentration of about 1 mM to about 50 mM, wherein the buffer is histidine buffer; surfactant at a concentration of about 0.01% to about 0.4%, wherein the surfactant is a non-ionic surfactant selected from the group comprising of poloxamer (e.g., poloxamer 188) or polyoxyethylene sorbitan fatty acid esters, e.g. polysorbate 20, polysorbate 40, polysorbate 60 or polysorbate 80; stabilizer- 1 at a concentration of about 10 mM to about 300 mM, wherein stabilizer- 1 is trehalose; and stabilizer-2 at a concentration of about 1 pM to about 1 mM, wherein stabilizer is pentetic acid.

[0041] In yet another embodiment of the present disclosure, the secukinumab formulation comprises buffer at a concentration of about 1 mM to about 50 mM, wherein the buffer is histidine buffer; surfactant at a concentration of about 0.01% to about 0.4%, wherein the surfactant is a non-ionic surfactant selected from the group comprising of poloxamer (e.g., poloxamer 188) or polyoxyethylene sorbitan fatty acid esters, e.g. polysorbate 20, polysorbate 40, polysorbate 60 or polysorbate 80; stabilizer- 1 at a concentration of about 10 mM to about 300 mM, wherein stabilizer- 1 is trehalose; and stabilizer-2 at a concentration of about 1 mM to about 100 mM, wherein stabilizer-2 is phenylalanine and / or pharmaceutically acceptable salts thereof.

[0042] In a preferred embodiment, formulation of the present invention comprises of 150 mg / ml of secukinumab; 20 mm of histidine buffer; 10 mM to 50 mM of phenylalanine; 200 mM trehalose; 0.02% polysorbate 20; water for injection; and pH 5.8 ± 0.5. In a more preferred embodiment, formulation of the present invention comprises of 150 mg / ml of secukinumab; 20 mm of histidine buffer; 37 mM of phenylalanine; 200 mM trehalose; 0.02% polysorbate 20; water for injection; and pH 5.8 ± 0.5.

[0043] In yet another embodiment of the present disclosure, the secukinumab formulation comprises of buffer at a concentration of about 1 mM to about 50 mM, wherein the buffer is histidine buffer oracetate buffer; surfactant at a concentration of about 0.01% to about 0.4%, wherein the surfactant is a non-ionic surfactant selected from the group comprising of poloxamer (e.g., poloxamer 188) or polyoxyethylene sorbitan fatty acid esters, e.g. polysorbate 20, polysorbate 40, polysorbate 60 or polysorbate 80; stabilizer- 1 at a concentration of about 10 mM to about 300 mM, wherein stabilizer- 1 is trehalose and stabilizer-2 at a concentration of about 1 mM to about 100 mM, wherein stabilizer-2 is pyridoxine and / or its pharmaceutically acceptable salts such as pyridoxine hydrochloride.

[0044] In yet another embodiment of the present disclosure, the secukinumab formulation comprises of buffer at a concentration of about 1 mM to about 50 mM, wherein the buffer is acetate buffer or histidine buffer; surfactant at a concentration of about 0.01% to about 0.4%, wherein the surfactant is a non-ionic surfactant selected from the group comprising of poloxamer (e.g., poloxamer 188) or polyoxyethylene sorbitan fatty acid esters, e.g. polysorbate 20, polysorbate 40, polysorbate 60 or polysorbate 80; stabilizer- 1 at a concentration of about 10 mM to about 300 mM, wherein stabilizer- 1 is trehalose or sorbitol; and stabilizer-2 at a concentration of about 1 mM to about 100 mM, wherein stabilizer-2 is proline and / or pharmaceutically acceptable salts thereof.

[0045] In another preferred embodiment, formulation of the present invention comprises of 150 mg / ml of secukinumab; 20 mM of acetate buffer; 10 mM to 50 mM of proline; 200 mM trehalose; 0.02% polysorbate 20; water for injection; and pH 5.8 ± 0.5. In a more preferred embodiment, formulation of the present invention comprises of 150 mg / ml of secukinumab; 20 mM of acetate buffer; 25 mM of proline; 200 mM trehalose; 0.02% polysorbate 20; water for injection; and pH 5.8 ± 0.5. In yet another preferred embodiment, formulation of the present invention comprises of 150 mg / ml of secukinumab; 20 mm of histidine buffer; 10 mM to 50 mM of proline; 200 mM trehalose; 0.02% polysorbate 20; water for injection; and pH 5.8 ± 0.5. In a more preferred embodiment, formulation of the present invention comprises of 150 mg / ml of secukinumab; 20 mM of histidine buffer; 25 mM of proline; 200 mM trehalose; 0.02% polysorbate 20; water for injection; and pH 5.8 ± 0.5. The formulation were prepared as per any of the embodiments of the present invention, stored and analysed for various pharmaceutical quality attributes of injectable formulation like pH, osmolality, protein content, free thiol ratio, low aggregates, High Molecular Weight species (% HMW), % Monomer using Size-Exclusion High-Performance Liquid Chromatography (SE-HPLC), Total Low Molecular Weight species (% LMW) using Capillary Electrophoresis-Sodium Dodecyl Sulfate (CE-SDS (NR)), Variants (acidic / main / basic ) were analysed using Ion Exchange - High-Performance Liquid Chromatography (IEX-HPLC), and Melting Temperature (Tm) analysis using Differential Scanning Fluorimetry (DSF)]. Capillary electrophoresis as perthe present embodiment may be carried out using Beckman Coulter (AB Sciex) PA-800 plus / CESI 8000 plus as per standard operating procedure provided by the equipment manufacturer. High performance liquid chromatography (HPLC) as per the present embodiment may be carried out using Arc HPLC system with PDA detector as per standard operating procedure provided by the equipment manufacturer.

[0046] Examples:

[0047] Example 1: Selection of excipients

[0048] Based on initial formulation excipients screening studies, phenylalanine and proline were selected for formulation stability studies, and further its stability effect on secukinumab (active molecule) were assessed.

[0049] Other excipients like buffer(s), surfactant(s) and solvents for the formulation were selected based on the pharmacopeial recommendations and achieving desired quality attributes for injectable formulation like pH, osmolality, protein content, free thiol ratio, low aggregates.

[0050] Surfactants like Polysorbate 20 and polysorbate 80 were found to be effective in controlling aggregation of secukinumab in formulation .

[0051] Buffers like Histidine buffer and Acetate buffer were found to be effective in stabilizing secukinumab in formulation.

[0052] Additional stabilizers like sugars and / or sugar alcohols like sucrose, trehalose, sorbitol, xylitol, etc. were found to be effective in providing additional stabilization effect on secukinumab in formulation.

[0053] Example 2: Optimization studies for stabilizer concentrations

[0054] Varied concentration of proline and phenylalanine were studied & compared for stability parameters under stress conditions [40 °C] to identify the effective range concentrations of stabilizers, and to study its effect on improvement of formulation stability.

[0055] Table 1

[0056]

[0057]

[0058] Results:

[0059] Effect of above formulations were studied under stress conditions [40 °C] for 2 weeks for stability [High Molecular Weight species (% HMW), % Monomer using Size-Exclusion High-Performance Liquid Chromatography (SE-HPLC) and Melting Temperature (Tm) analysis using Differential Scanning Fluorimetry (DSF)].

[0060] Table 2

[0061]

[0062] % HMW and % Monomers was found to be within desired limits in all formulations excluding control; the concentration variation had minimal impact on aggregation and Tm and thus all formulations comprising proline or phenylalanine studied as stabilizer were found to be stable.

[0063] Example 3: Formulation Design

[0064] Below mentioned formulation were prepared in 1 ml solution as per standard pharmaceutical procedures, and further were tested for quality attributes like pH, osmolality, protein content, and free thiol ratio. [F0 was used as control]

[0065] Table 3.1

[0066]

[0067]

[0068] Table 3.2

[0069]

[0070] Results:

[0071] Quality attributes of prepared formulation like pH, Osmolality, Protein Content, Free Thiol ratio and Potency of Fl, F2, F3, F4, F5, and F6 were found to be within pharmaceutically acceptable limits as shown below in table 4.

[0072] Table 4

[0073]

[0074] Example 4 : Stability studies for Fl formulation

[0075] The formulation Fl (prepared as detailed in Example 3) was studied for stability parameters including % HMW, % Monomer were analysed using SE-HPLC, Total Low Molecular Weight species (% LMW) using Capillary Electrophoresis- Sodium Dodecyl Sulfate (CE-SDS (NR)), Variants (acidic / main / basic ) were analysed using Ion Exchange - High-Performance Liquid Chromatography (IEX-HPLC), and Tm analysis was performed using Differential Scanning Fluorimetry (DSF).

[0076] Results obtained were as follows:

[0077] • Stress conditions ( 40 °C) at interval of 2 weeks

[0078] Table 5

[0079]

[0080] Table 6

[0081]

[0082] • Accelerated time conditions ( 25 ± 2 °C) at interval of 1 month for 3 months

[0083] Table 7

[0084]

[0085] Table 8

[0086]

[0087]

[0088] • Real time conditions (5 ± 3 °C) at interval of 1 month for 3 months

[0089] Table 9

[0090]

[0091] Table 10

[0092]

[0093] The Formulation Fl was found to be stable for stability parameters including %HMW, % Monomer using SE-HPLC, % Total LMW using CE-SDS (NR), Variants analysis (acidic / main / basic ) using IEX-HPLC, and Tm analysis using Differential Scanning Fluorimetry (DSF). [Refer table 5 - 10; Fig. 1(a), Fig. 1(b), Fig. 1(c), Fig. 2(a), Fig. 2(b) and Fig. 2(c)],

[0094] Example 5 : Stability studies for F2 formulation

[0095] The formulation F2 (prepared as detailed in Example 3) was studied for stability parameters including % HMW, % Monomer were analysed using SE-HPLC, % Total LMW using CE-SDS (NR), Variants (acidic / main / basic) were analysed using IEX-HPLC, and Tm analysis was performed using Differential Scanning Fluorimetry (DSF).

[0096] Results obtained were as follows:

[0097] • Stress conditions ( 40 °C) at interval of 2 weeks

[0098] Table 11

[0099]

[0100]

[0101] Table 12

[0102]

[0103] • Accelerated time conditions ( 25 ± 2 °C) at interval of 1 month for 3 months Table 13

[0104]

[0105] Table 14

[0106]

[0107] • Real time conditions (5 ± 3 °C) at interval of 1 month for 3 months

[0108] Table 15

[0109]

[0110] Table 16

[0111]

[0112] The Formulation F2 was found to be stable for stability parameters including %HMW, % Monomer using SE-HPLC, % Total LMW using CE-SDS (NR), Variants analysis (acidic / main / basic ) using IEX-HPLC, and Tm analysis using Differential Scanning Fluorimetry (DSF). [Refer table 11-16; Fig. 1(a), Fig. 1(b), Fig. 1(c), Fig. 2(a), Fig. 2(b) and Fig. 2(c)]

[0113] Example 6 : Stability studies for F3 formulation

[0114] The formulation F3 (prepared as detailed in Example 3) was studied for stability parameters including % HMW, % Monomer were analysed using SE-HPLC, % Total LMW using CE-SDS (NR), Variants (acidic / main / basic) were analysed using IEX-HPLC, and Tm analysis was performed using Differential Scanning Fluorimetry (DSF).

[0115] Results obtained were as follows:

[0116] • Stress conditions ( 40 °C) at interval of 2 weeks

[0117] Table 17

[0118]

[0119] Table 18

[0120]

[0121] • Accelerated time conditions ( 25 ± 2 °C) at interval of 1 month for 3 monthsTable 19

[0122]

[0123] Table 20

[0124]

[0125] • Real time conditions (5 ± 3 °C) at interval of 1 month for 3 months

[0126] Table 21

[0127]

[0128] Table 22

[0129]

[0130] The Formulation F3 was found to be stable for stability parameters including %HMW, % Monomer using SE-HPLC, % Total LMW using CE-SDS (NR), Variants analysis (acidic / main / basic ) using IEX-HPLC, and Tm analysis using Differential Scanning Fluorimetry (DSF). [Refer table 17 - 22; Fig. 1(a), Fig. 1(b), Fig. 1(c), Fig. 2(a), Fig. 2(b) and Fig. 2(c)]Example 7 : Stability studies for F4 formulation

[0131] The formulation F4 (prepared as detailed in Example 3) was studied for stability parameters including % HMW, % Monomer were analysed using SE-HPLC, and Tm analysis was performed using Differential Scanning Fluorimetry (DSF).

[0132] Results obtained were as follows:

[0133] • Stress conditions ( 40 °C) at interval of 2 weeks

[0134] Table 23

[0135]

[0136] The Formulation F4 was found to be stable for stability parameters including %HMW, % Monomer using SE-HPLC, and Tm analysis using Differential Scanning Fluorimetry (DSF).

[0137] [Refer table 23]

[0138] Example 8 : Stability studies for F5 formulation

[0139] The formulation F5 (prepared as detailed in Example 3) was studied for stability parameters including % HMW, % Monomer were analysed using SE-HPLC, and Tm analysis was performed using Differential Scanning Fluorimetry (DSF).

[0140] Results obtained were as follows:

[0141] • Stress conditions ( 40 °C) at interval of 2 weeks

[0142] Table 24

[0143]

[0144] The Formulation F5 was found to be stable for stability parameters including %HMW, % Monomer using SE-HPLC, and Tm analysis using Differential Scanning Fluorimetry (DSF).

[0145] [Refer table 24]Example 9 : Stability studies for F6 formulation

[0146] The formulation F6 (prepared as detailed in Example 3) was studied for stability parameters including % HMW, % Monomer were analysed using SE-HPLC, and Tm analysis was performed using Differential Scanning Fluorimetry (DSF).

[0147] Results obtained were as follows:

[0148] • Stress conditions ( 40 °C) at interval of 2 weeks

[0149] Table 25

[0150]

[0151] The Formulation F6 was found to be stable for stability parameters including %HMW, % Monomer using SE-HPLC, and Tm analysis using Differential Scanning Fluorimetry (DSF).

[0152] [Refer table 25]

[0153] Although the present disclosure may be susceptible to different embodiments, and the present disclosure can be considered an exemplification of the principles of the disclosure and is not intended to limit the scope of disclosure to which it is illustrated and disclosed in this description. Embodiments are provided so as to thoroughly and fully convey the scope of the present disclosure to the person skilled in the art. Numerous details are set forth, relating to specific components, and processes, to provide a complete understanding of embodiments of the present disclosure. It will be evident to the person skilled in the art that the details provided in the embodiments should not be construed to limit the scope of the present disclosure. In some embodiments, well-known composition, well-known processes, and well-known techniques are not described in detail. The terminology used, in the present disclosure, is only for the purpose of explaining a particular embodiment and such terminology shall not be considered to limit the scope of the present disclosure. The particular order of steps disclosed in the process of the present disclosure is not to be construed as necessarily requiring their performance as described or illustrated. It is also to be understood that additional or alternative steps may be employed.

Claims

Claims:

1. A liquid formulation comprisinga. secukinumab;b. optionally, one or more buffer(s);c. one or more surfactant(s);d. first stabilizer selected from group comprising of sugar(s), sugar alcohol, and combinations thereof; ande. second stabilizer selected from group comprising of pentetic acid, pyridoxine, proline, phenylalanine, and combinations thereof.

2. The formulation as claimed in claim 1, wherein secukinumab is at a concentration of about 25 mg / ml to about 300 mg / ml.

3. The formulation as claimed in claim 1, wherein one or more buffer(s) is at a concentration of about ImM to about 50 mM and is selected from group comprising of histidine buffer, citrate buffer, acetate buffer, phosphate buffer, tris buffer, and combinations thereof.

4. The formulation as claimed in claim 1, wherein one or more surfactant(s) is selected from group comprising of poloxamer and polyoxyethylene sorbitan fatty acid esters, and combinations thereof; and wherein concentration of surfactant is about 0.01% to about 0.4%.

5. The formulation as claimed in claim 1, wherein first stabilizer is selected from group comprising of sugar(s) and sugar alcohol including sucrose, trehalose, raffinose, maltose, sorbitol, xylitol, mannitol, and combinations thereof; and wherein concentration of first stabilizer is about 10 mM to about 300 mM.

6. The formulation as claimed in claim 1, wherein concentration of second stabilizer is about 1 pM to about 100 mM.

7. The formulation as claimed in claim 1, wherein the formulation is devoid of methionine.

8. The formulation as claimed in claim 1, wherein the formulation comprises:i. 25 mg / ml to 300 mg / ml of secukinumab;ii. at least one buffer(s) selected from histidine buffer and acetate buffer, at a concentration of about 1 mM to about 50 mM;iii. at least one stabilize^ s) selected from proline and phenylalanine, at a concentration of about 1 mM to 100 mM;iv. at least one sugar selected from sucrose and trehalose, at a concentration of about 10 mM to 300 mM;v. a surfactant selected from pol oxamer 188, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, and combinations thereof at a concentration of about 0.01% to about 0.4%; andvi . water for inj ection.

9. A liquid formulation comprising :i. 150 mg / ml of secukinumab;ii. about 10 mm to about 30 mm of histidine buffer or acetate buffer;iii. about 10 mm to about 50 mm of proline or phenylalanine;iv. about 200 mm trehalose;v. about 0.01% to about 0.4% polysorbate 20;vi. water for injection and pH 5.8 ± 0.5.

10. The formulation of claim 9, wherein the concentration of histidine buffer or acetate buffer is about 20 mM.

11. The formulation of claim 9, wherein the concentration of polysorbate 20 is about 0.02%.

12. The formulation of claim 9, wherein the proline is at a concentration of about 25 mM.

13. The formulation of claim 9, wherein the phenylalanine is at a concentration of about 37 mM.

14. The formulation as claimed in claim 9, wherein the formulation is devoid of methionine.

15. The formulation of claim 9, wherein the formulation comprises ofi. 150 mg / ml of secukinumab;ii. 20 mM of histidine buffer or acetate buffer;iii. about 25 mM proline or about 37 mM phenylalanine;iv. 200 mm trehalose; v. 0.2% polysorbate 20; vi. water for injection; and vii. pH 5.8 ± 0.5.

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