Anti-IL-13R antibody preparations

Abstract

A stable formulation of an anti-IL13R antibody. Also provided are uses of the formulation for therapy, methods of therapy using the formulation, and processes for producing the formulation.

Description

[Technical field]

[0001] The present disclosure relates to formulations of anti-IL13R antibodies and uses thereof, methods of treatment using said formulations, particularly for treating the conditions disclosed herein, as well as processes for producing said formulations. [Background technology]

[0002] IL-13 has been implicated in a variety of pathologies, including, but not limited to, various respiratory and allergic disorders, fibrosis, scleroderma, atopic dermatitis, inflammatory bowel disease, and certain cancers; see, for example, Wynn, TA, 2003 Annu.Rev.Immunol.21:425-456; Terabe et al., 2000 Nat.Immunol.1(6):515-520; Fuss et al., 2004 J.Clin.Invest.113(10):1490-1497; Simms et al., 2002 Curr.Opin.Rheumatol.14(6):717-722; and Hasegawa et al., 1997 J.Rheumatol.24(2):328-332. Thus, IL-13 is an attractive target for the treatment of such diseases.

[0003] One possible means of inhibiting the activity of IL-13 would be to interfere with the binding of IL-13 to its receptor IL-13R, for example by using an antibody specific for IL-13R, such as an antibody specific for IL-13Rα1. An effective antibody antagonist against IL-13Rα1 could interfere with the binding of IL-13 and prevent the heterodimerization between IL-4Rα and IL-13Rα1. Such an antibody would inhibit both IL-13 and IL-4 signaling through the type II receptor, while retaining IL-4 signaling through the type I receptor. Signaling through the type I receptor is essential for the induction phase of the immune response during which Th2 cells differentiate. Since T cells do not express IL-13Rα1, the type II receptor plays no role in Th2 differentiation. Therefore, IL-13Rα1 antibodies should not affect the overall Thl / Th2 balance. Signaling through the type II IL-4 / IL-13 receptor is important during the effector A stage of the immune response during established allergic inflammation. Thus, inhibition of the type II receptor should have beneficial effects on many of the symptoms of asthma and other IL-13R-mediated pathologies and would therefore be an effective disease-modifying drug.

[0004] Antibodies (both monoclonal and polyclonal) against IL-13Rα1 have been reported in the art; see, e.g., WO97 / 15663, WO03 / 80675; WO03 / 46009; WO06 / 072564; Gauchat et al., 1998 Eur. J. Immunol. 28:4286-4298; Gauchat et al., 2000 Eur. J. Immunol. 30:3157-3164; Clement et al., 1997 Cytokine 9(11):959(Meeting Abstract); Ogata et al., 1998 J. Biol. Chem. 273:9864-9871; Graber et al., 1998 Eur. J. Immunol. 28:4286-4298; C. Vermot-Desroches et al., 2000 Tissue See Antigens 5(Supp.l):52-53(Meeting Abstract); Poudrier et al., 2000 Eur. J. Immunol. 30:3157-3164; Akaiwa et al., 2001 Cytokine 13:75-84; Cancino-Diaz et al., 2002 J. Invest. Dermatol. 119:1114-1120; and Krause et al., 2006 MoI. Immunol. 43:1799-1807.

[0005] One particularly promising anti-IL-13Rα1 antibody is CSL334 (now called ASLAN004 / Ebrasakimab), described as antibody 10G5-6 in WO2008 / 060813. Ebrasakimab has been shown to bind to human IL-13Rα1 with high affinity (e.g., Kd may be as low as 500pM). Ebrasakimab has been shown to effectively antagonize IL-13 function by inhibiting the binding of IL-13 to its receptor IL-13Rα1, and inhibits IL-13- and IL-4-induced eotaxin release in NHDF cells, IL-13- and IL-4-induced STAT6 phosphorylation in NHDF cells, and IL-13-stimulated release of TARC in blood or peripheral blood mononuclear cells.

[0006] However, an optimized formulation of ebrasakimab is needed because it can be difficult to handle and manufacture. Creating a highly concentrated antibody formulation, for example with an antibody / fragment concentration of more than 150 mg / ml, would be beneficial for patients as it would allow for smaller injection volumes, but is not easy to achieve. The formation of aggregates and particles in parenteral formulations is very dangerous for patients and must be avoided. Therefore, an optimized formulation is needed to address these issues and / or to maximize the shelf life, delivery, potency, and efficacy of the formulation. Summary of the Invention

[0007] The present disclosure is summarized in the following items. [Item 1] 1. A highly concentrated antibody formulation, the formulation comprising: 150-210 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof, for example 150, 155, 160, 165, 170, 175, 180, 185, 190, 195 or 200 mg / ml, in particular 150 mg / ml, 175 mg / ml or 200 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof; 170-250 mM arginine (such as Arg-HCl or Arg-Glu), for example 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245 or 250 mM, in particular 150 mM, 175 mM or 250 mM arginine; 20-50 mM histidine buffer, for example 20, 25, 30, 35, 40, 45 or 50 mM (such as 20 mM or 50 mM) histidine buffer; 0.01-0.03% non-ionic surfactant, for example 0.01-0.03% w / w (such as 0.02% w / w) non-ionic surfactant; wherein the pH of the formulation is in the range of 6.0 to 7.0 (such as in the range of 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9 or 7.0), in particular pH 6.5; and wherein the anti-IL-13R antibody or antigen-binding fragment thereof comprises a VH CDR1 comprising the amino acid sequence as set forth in SEQ ID NO:1, a VH CDR2 comprising the amino acid sequence as set forth in SEQ ID NO:2, and a VH CDR3 comprising the amino acid sequence as set forth in SEQ ID NO:3; and a VL CDR1 comprising the amino acid sequence as set forth in SEQ ID NO:4, a VL CDR2 comprising the amino acid sequence as set forth in SEQ ID NO:5, and a VL CDR3 comprising the amino acid sequence as set forth in SEQ ID NO:6. Highly concentrated antibody preparation. [Item 2] 2. The formulation according to item 1, wherein the formulation comprises 150-200 mg / ml (such as 150, 155, 160, 165, 170, 175, 180, 185, 190, 195 or 200 mg / ml) of an anti-IL13R antibody or binding fragment thereof. [Item 3] 3. A formulation according to item 1 or 2, wherein the formulation comprises 150, 175 or 200 mg / ml of the antibody. [Item 4] A formulation according to item 3, containing 150 mg / ml. [Item 5] A formulation according to item 3, containing 175 mg / ml. [Item 6] A formulation according to item 3, containing 200 mg / ml. [Item 7] 7. A formulation according to any one of items 1 to 6, wherein the formulation comprises 175, 200 or 250 mM arginine. [Item 8] 8. A formulation according to any one of items 1 to 7, wherein arginine is Arg-HCl. [Item 9] 9. A formulation according to any one of items 1 to 8, wherein the formulation comprises a 20 mM histidine buffer. [Item 10] 10. A formulation according to any one of items 1 to 9, wherein the formulation further comprises phenylalanine (such as 45 to 85 mM phenylalanine). [Item 11] 11. The formulation according to item 10, wherein the formulation comprises 50, 75 or 80 mM phenylalanine. [Item 12] 12. The formulation according to any one of items 1 to 11, wherein the formulation further comprises CaCl2 (e.g., 30, 35, 40, 45, 50, 55, 60, 65 or 70 mM CaCl2). [Item 13] 13. A formulation according to item 12, wherein the formulation contains 50 mM CaCl2. [Item 14] 14. A formulation according to any one of items 1 to 13, wherein the osmolality of the formulation is in the range of 350 to 550 mOsmo / kg (e.g. 350, 355, 365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495, 500, 505, 515, 520, 525, 530, 535, 540, 545, 550 mOsmo / kg (such as 405 to 435 mOsmo / kg)). [Item 15] 15. A formulation according to any one of items 1 to 14, further comprising 50 to 200 mM sugar (e.g. 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200 mM sugar (such as 180 mM sugar)). [Item 16] 16. A formulation according to item 15, wherein the formulation comprises 180 mM sugar. [Item 17] 17. The formulation according to item 15 or 16, wherein the sugar is selected from mannitol, sorbitol, dextrose, galactose, fructose, lactose, trehalose and sucrose. [Item 18] 18. A formulation according to item 17, wherein the sugar is sucrose. [Item 19] A formulation according to any one of items 1 to 18 comprising 0.02% w / w of a non-ionic surfactant. [Item 20] A formulation according to any one of items 1 to 19, wherein the non-ionic surfactant is a polysorbate, e.g. polysorbate 20, 40, 60, or 80, polysorbate 20. [Item 21] 21. The formulation according to item 20, wherein the non-ionic surfactant is polysorbate 20. [Item 22] 22. A formulation according to any one of items 1 to 21, wherein the formulation does not contain NaCl. [Item 23] 22. The formulation according to any one of items 1 to 21, wherein the formulation comprises 50 to 150 mM NaCl (e.g. 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150 mM NaCl (such as 62.5 or 140 mM NaCl)). [Item 24] 24. Formulation according to any one of items 1 to 23, wherein the formulation has a viscosity in the range of 10 to 30 cP (mPa.s), such as 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 cP (such as 15 to 25 cP, in particular 20 cP). [Item 25] 25. The formulation according to any one of items 1 to 24, wherein the anti-IL-13R antibody is an anti-IL13Rα1 antibody. [Item 26] 26. The formulation according to any one of items 1 to 25, wherein the anti-IL-13R antibody binds to the epitope FFYQ. [Item 27] 27. The formulation according to any one of items 1 to 26, wherein the anti-IL-13R antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO:7, or a sequence at least 95% identical thereto. [Item 28] 28. The formulation according to any one of items 1 to 27, wherein the anti-IL-13R antibody comprises a VL domain comprising the amino acid sequence shown in SEQ ID NO:8 or a sequence at least 95% identical thereto. [Item 29] 29. The formulation according to any one of items 1 to 28, wherein the anti-IL13R antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 7, or a sequence at least 95% identical thereto, and a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 8, or a sequence at least 95% identical thereto. [Item 30] 30. The formulation according to any one of items 1 to 29, wherein the anti-IL13R antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO:7 and a VL domain comprising the amino acid sequence set forth in SEQ ID NO:8. [Item 31] The formulation according to any one of items 1 to 30, wherein the anti-IL13R antibody is a human antibody. [Item 32] 32. The formulation according to any one of items 1 to 31, wherein the formulation comprises 175 mg / ml of anti-IL-13R antibody or antigen-binding fragment thereof; 250 mM arginine; 20 mM histidine buffer; 0.02% non-ionic surfactant, and wherein the pH of the formulation is 6.5. [Item 33] 33. The formulation according to any one of items 1 to 32, comprising: 175 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof; 250 mM Arg-HCl; 20 mM histidine buffer; 0.02% polysorbate 20, and wherein the pH of the formulation is 6.5. [Item 34] 34. The formulation according to any one of items 1 to 33, comprising 175 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof; 250 mM Arg-HCl; 20 mM histidine buffer; 0.02% polysorbate 20; phenylalanine (such as 45 to 85 mM phenylalanine), and wherein the pH of the formulation is 6.5. [Item 35] 35. The formulation according to any one of items 1 to 34, comprising: 175 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof; 250 mM Arg-HCl; 20 mM histidine buffer; 0.02% polysorbate 20; 75 mM phenylalanine, and wherein the pH of the formulation is 6.5. [Item 36] 36. The formulation according to any one of items 1 to 35, comprising 175 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof, wherein the anti-IL13R antibody comprises a VL domain comprising the amino acid sequence set forth in SEQ ID NO:8, or a sequence at least 95% identical thereto, and a VH domain comprising the amino acid sequence set forth in SEQ ID NO:7, or a sequence at least 95% identical thereto; the formulation comprises 250 mM Arg-HCl; 20 mM histidine buffer; 0.02% polysorbate 20; 75 mM phenylalanine, and wherein the pH of the formulation is 6.5. [Item 37] 37. A formulation according to any one of items 1 to 36 for use in therapy. [Item 38] 38. A formulation according to item 37 for use in the treatment of an inflammatory or autoimmune disease, such as chronic inflammation. [Item 39] 39. The formulation according to item 38, wherein the inflammation is selected from the group comprising fibrosis (including pulmonary fibrosis (such as cystic fibrosis, idiopathic pulmonary fibrosis, progressive widespread fibrosis); liver fibrosis (such as liver cirrhosis); heart diseases (such as atrial fibrosis, endomyocardial fibrosis, old myocardial infarction); articular fibrosis; Dupuytren's contracture; keloid fibrosis; mediastinal fibrosis; myelofibrosis; nephrogenic systemic fibrosis; retroperitoneal fibrosis; and scleroderma), Hodgkin's disease, ulcerative colitis, Crohn's disease, atopic dermatitis, eosinophilic esophagitis, allergic rhinitis, asthma and chronic lung diseases (including chronic obstructive pulmonary disease), in particular asthma. [Item 40] 37. Use of a formulation according to any one of items 1 to 36 for use in the manufacture of a therapeutic medicament for the treatment of an inflammatory or autoimmune disease, such as chronic inflammation. [Item 41] 40. Use of a formulation according to item 40 in the manufacture of a therapeutic medicament for the treatment of a condition selected from the group consisting of fibrosis (including pulmonary fibrosis (such as cystic fibrosis, idiopathic pulmonary fibrosis, progressive widespread fibrosis); liver fibrosis (such as liver cirrhosis); heart diseases (such as atrial fibrosis, endomyocardial fibrosis, old myocardial infarction); articular fibrosis; Dupuytren's contracture; keloid fibrosis; mediastinal fibrosis; myelofibrosis; nephrogenic systemic fibrosis; retroperitoneal fibrosis; and scleroderma), Hodgkin's disease, ulcerative colitis, Crohn's disease, atopic dermatitis, eosinophilic esophagitis, allergic rhinitis, asthma and chronic lung diseases (including chronic obstructive pulmonary disease), Sézary syndrome, in particular asthma. [Item 42] 37. A method of treatment comprising administering an effective amount of a formulation according to any one of items 1 to 36. [Item 43] A method for treating inflammation (such as chronic inflammation) or an autoimmune disease, comprising administering an effective amount of a formulation according to any one of items 1 to 36. [Item 44] A method according to item 43, comprising: Fibrosis (pulmonary fibrosis (such as cystic fibrosis, idiopathic pulmonary fibrosis, progressive widespread fibrosis); liver fibrosis (such as cirrhosis); heart disease (such as atrial fibrosis, endomyocardial fibrosis, old myocardial infarction); articular fibrosis; Dupuytren's contracture; keloid fibrosis; mediastinal fibrosis; myelofibrosis; nephrogenic systemic fibrosis; retroperitoneal fibrosis; and scleroderma). Treatment of a condition selected from the group including Hodgkin's disease, ulcerative colitis, Crohn's disease, atopic dermatitis, eosinophilic esophagitis, allergic rhinitis, asthma and chronic lung disease (including chronic obstructive pulmonary disease), in particular asthma.

[0008] Antibodies such as Ebrasakimab need to be formulated at high concentrations to allow the desired dose in humans to be administered in the smallest possible volume. High concentration formulations pose inherent challenges since phase separation-like phenomena can be observed. Aggregation is also a common feature with high concentration antibodies. However, the formulation needs to contain very high levels of antibody molecules as "monomers" (e.g., 99% or more monomers). Furthermore, the formulation needs to be stable upon storage. Ebrasakimab appears to have a hydrophobic moiety in its protein; it interacts, for example, with hydrophobic interaction columns in the absence of high salt concentrations. This hypothetical hydrophobic moiety adds an additional complication when formulating the antibody while preventing aggregation. Thus, the antibodies of the present disclosure are particularly difficult to formulate.

[0009] Furthermore, a challenging problem presented by more concentrated antibody formulations is the tendency of such formulations to be excessively viscous, and therefore the formulation needs to be optimized to keep the viscosity at an acceptably low level, i.e., close to the target viscosity range of 20-25 cP (e.g., 20 cP).

[0010] The inventors have optimized the formulations of the present disclosure and established that IL-13R antibodies (such as ebrasakimab) are most suitable for formulation within a narrow set of parameters. The formulations of the present disclosure are highly monomeric, e.g., at least 98% monomeric (e.g., 98-99.5% monomeric), even when formulated at high antibody concentrations. The formulations of the present disclosure also have good viscosity at high antibody concentrations. Furthermore, the formulations are suitably stable, e.g., in some embodiments, when stored at 4° C. or 25° C. for 90 days, there is no change in monomer content or less than a 0.5% decrease in monomer. Accelerated "stress testing" at 40° C. also shows that the formulations of the present disclosure are stable over a 60-day period, e.g., using potency measurements.

[0011] A combination of characteristics of the formulations of the present disclosure, including pH, contribute to stabilizing the IL-13 receptor antibody or binding fragment thereof.

[0012] In one embodiment, a formulation of the disclosure has a viscosity in the range of 10 to 30 cP (centipoise), such as 20 cP, e.g., 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 cP, e.g., at ambient temperature. Surprisingly, the viscosity of the formulation of the disclosure is relatively low, even at high antibody concentrations.

[0013] In one embodiment, the viscosity is measured using a viscometer such as a rotational viscometer, an electromagnetic ball rotational (EMS) viscometer, or a Stabinger viscometer. In one embodiment, the viscosity is measured using a rheometer such as a shear rheometer, a dynamic shear rheometer, an extensional rheometer, or a capillary rheometer. In one embodiment, the viscosity is measured using a Kinexus-ultra+ rheometer (Netzsch).

[0014] In one embodiment, the osmolality of the formulation is in the range 350-450 mOsmo / kg (such as 390-430 mOsmo / kg, in particular 410+ / -5 mOsmo / kg).

[0015] In one embodiment, the formulation comprises 150-210 mg / ml of anti-IL13R antibody, for example 150-175 mg / ml (such as 150, 155, 160, 165, 170, 175 mg / ml) of anti-IL13R antibody. In one embodiment, the formulation comprises 175 mg / ml to 210 mg / ml (such as 175, 180, 185, 190, 195, 200, 205 or 210 mg / ml). In one embodiment, the formulation comprises 150 mg / ml of anti-IL13R antibody. In another embodiment, the formulation comprises 175 mg / ml of anti-IL13R antibody. In one embodiment, the formulation comprises 200 mg / ml.

[0016] In one embodiment, the formulation comprises 170-260 mM arginine, such as 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250 or 260 mM arginine. In one embodiment, the formulation comprises 150 mM, 175 mM, 200 mM or 250 mM arginine. In one embodiment, the formulation comprises 150 mM arginine. In one embodiment, the formulation comprises 175 mM arginine. In one embodiment, the formulation comprises 200 mM arginine. In one embodiment, the formulation comprises 250 mM arginine.

[0017] In one embodiment, the arginine is Arg-HCl. In another embodiment, the arginine is Arg-Glu. In one embodiment, the arginine is L-arginine. Thus, in one embodiment, the formulation comprises 150 mM Arg-HCl. In one embodiment, the formulation comprises 175 mM Arg-HCl. In one embodiment, the formulation comprises 200 mM Arg-HCl. In one embodiment, the formulation comprises 250 mM Arg-HCl.

[0018] In one embodiment, the formulation comprises a 20-50 mM histidine buffer, such as a 20, 25, 30, 35, 40, 45 or 50 mM histidine buffer, such as 20 mM or 50 mM. In one embodiment, the formulation comprises a 20 mM histidine buffer. In another embodiment, the formulation comprises a 50 mM histidine buffer.

[0019] In one embodiment, the formulation comprises 0.01-0.03% non-ionic surfactant (especially 0.02%, such as 0.01, 0.015, 0.02, 0.025, or 0.030%). In one embodiment, the formulation comprises 0.01-0.03% (especially 0.02%, such as 0.01, 0.015, 0.02, 0.025, or 0.030%) volume / volume (v / v) non-ionic surfactant. In one embodiment, the formulation comprises 0.01-0.03% (especially 0.02%, such as 0.01, 0.015, 0.02, 0.025, or 0.030%) weight / volume (w / v) non-ionic surfactant. In one embodiment, the formulation comprises 0.01 to 0.03% (especially 0.02%) weight / weight (w / w) of non-ionic surfactant, such as 0.01, 0.015, 0.02, 0.025 or 0.030%. In one embodiment, the formulation comprises 0.02% w / w of non-ionic surfactant.

[0020] In one embodiment, the non-ionic surfactant is a polysorbate (such as polysorbate 20, 40, 60, or 80). In one embodiment, the non-ionic surfactant is polysorbate 20. Thus, in one embodiment, the formulation comprises 0.01-0.03% (such as 0.02%) polysorbate 20 (e.g., as % w / w, as % w / v, as % v / w, or as % v / v). In one embodiment, the formulation comprises 0.02% w / w polysorbate 20.

[0021] In one embodiment, the pH of the formulation is in the range of 6.0 to 7.0 (such as 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, or 7.0). In one embodiment, the pH is 6.0, 6.5 or 7.0. In one embodiment, the pH is 6.5.

[0022] In one embodiment, the formulation further comprises phenylalanine (such as 45-90 mM phenylalanine, for example 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90 mM phenylalanine). In one embodiment, the formulation comprises 50, 75, or 80 mM phenylalanine. Thus, in one embodiment, the formulation comprises 50 mM phenylalanine. In one embodiment, the formulation comprises 75 mM phenylalanine. In one embodiment, the formulation comprises 80 mM phenylalanine.

[0023] In one embodiment, the formulation further comprises CaCl2 (e.g., 10, 20, 30, 40, 50, or 60 mM CaCl2). In one embodiment, the formulation comprises 50 mM CaCl2.

[0024] In one embodiment, the formulation further comprises 50-200 mM sugar (such as 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 mM sugar). In one embodiment, the formulation comprises 180 mM sugar. In one embodiment, the sugar is selected from mannitol, sorbitol, dextrose, galactose, fructose, lactose, trehalose, and sucrose. In one embodiment, the sugar is sucrose. Thus, in one embodiment, the formulation comprises 180 mM sucrose.

[0025] In one embodiment, certain formulations of the present disclosure have less than 1% protein aggregation when stored for 90 days at a temperature ranging from, for example, 2-25°C.

[0026] The anti-IL13R antibody formulations of the present disclosure are particularly suitable for stable long-term storage of anti-IL13R antibodies. In one embodiment, the formulation is stored at a temperature in the range of 2-8° C. (such as 2, 3, 4, 5, 6, 7, or 8° C., such as 4° C.).

[0027] In one embodiment, a parenteral formulation (particularly a liquid formulation) is provided, for example, for infusion or injection. In one embodiment, a parenteral liquid formulation is provided as a concentrate for dilution with a liquid for injection, such as glucose, saline or water for injection. In one embodiment, a parenteral liquid formulation is provided at a final concentration for administration without dilution, for example, for injection or infusion.

[0028] In one embodiment, the formulation comprises 175 mg / ml anti-IL-13R antibody or antigen-binding fragment thereof; 250 mM arginine; 20 mM histidine buffer; 0.02% non-ionic surfactant, and wherein the pH of the formulation is 6.5.

[0029] In one embodiment, the formulation comprises 175 mg / ml anti-IL-13R antibody or antigen-binding fragment thereof; 250 mM Arg-HCl; 20 mM histidine buffer; 0.02% polysorbate 20, and wherein the pH of the formulation is 6.5.

[0030] In one embodiment, the formulation comprises 175 mg / ml anti-IL-13R antibody or antigen-binding fragment thereof; 250 mM arginine; 20 mM histidine buffer; 0.02% non-ionic surfactant; phenylalanine (such as 45-85 mM phenylalanine), and wherein the pH of the formulation is 6.5.

[0031] In one embodiment, the formulation comprises 175 mg / ml anti-IL-13R antibody or antigen-binding fragment thereof; 250 mM arginine; 20 mM histidine buffer; 0.02% non-ionic surfactant; 75 mM phenylalanine, and wherein the pH of the formulation is 6.5.

[0032] In one embodiment, the formulation comprises 175 mg / ml anti-IL-13R antibody or antigen-binding fragment thereof; 250 mM Arg-HCl; 20 mM histidine buffer; 0.02% polysorbate 20; 75 mM phenylalanine, and wherein the pH of the formulation is 6.5.

[0033] In one embodiment, the antibodies or binding fragments used in the formulations of the present disclosure are monoclonal.

[0034] In one embodiment, the antibodies or binding fragments used in the formulations of the present disclosure are human. In one embodiment, the antibodies or binding fragments used in the formulations of the present disclosure are chimeric or humanized.

[0035] In one embodiment, the antibody or binding fragment thereof comprises a heavy chain variable region comprising a CDRH having the sequence set forth in SEQ ID NO:1, a CDRH2 having the sequence set forth in SEQ ID NO:2, and a CDRH3 having the sequence set forth in SEQ ID NO:3; and a light chain variable region comprising a CDRL1 having the sequence set forth in SEQ ID NO:4, a CDRL2 having the sequence set forth in SEQ ID NO:5, and a CDRL3 having the sequence set forth in SEQ ID NO:6.

[0036] In one embodiment, the antibody or binding fragment thereof comprises a VH domain comprising the sequence shown in SEQ ID NO: 7, or a sequence at least 95% identical thereto. In one embodiment, the antibody or binding fragment thereof comprises a VL domain comprising the sequence shown in SEQ ID NO: 8, or a sequence at least 95% identical thereto.

[0037] In one embodiment, the antibody or binding fragment thereof comprises a VH domain comprising the sequence shown in SEQ ID NO:7 or a sequence at least 95% identical thereto, and a VL domain comprising the sequence shown in SEQ ID NO:8 or a sequence at least 95% identical thereto.

[0038] In one embodiment, the antibody or binding fragment thereof comprises a VH domain comprising the sequence shown in SEQ ID NO:7 and a VL domain comprising the sequence shown in SEQ ID NO:8.

[0039] In one embodiment, the antibody or binding fragment thereof is ebrasakimab.

[0040] In one embodiment, the formulation comprises 175 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof comprising a VH domain comprising the sequence set forth in SEQ ID NO:7, or a sequence at least 95% identical thereto, and a VL domain comprising the sequence set forth in SEQ ID NO:8, or a sequence at least 95% identical thereto; 250 mM arginine; 20 mM histidine buffer; 0.02% non-ionic surfactant, and wherein the pH of the formulation is 6.5.

[0041] In one embodiment, the formulation comprises 175 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof comprising a VH domain comprising the sequence set forth in SEQ ID NO:7, or a sequence at least 95% identical thereto, and a VL domain comprising the sequence set forth in SEQ ID NO:8, or a sequence at least 95% identical thereto; 250 mM Arg-HCl; 20 mM histidine buffer; 0.02% polysorbate 20, and wherein the pH of the formulation is 6.5.

[0042] In one embodiment, the formulation comprises 175 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof comprising a VH domain comprising the sequence set forth in SEQ ID NO:7, or a sequence at least 95% identical thereto, and a VL domain comprising the sequence set forth in SEQ ID NO:8; 250 mM arginine; 20 mM histidine buffer; 0.02% non-ionic surfactant; phenylalanine (such as 45-85 mM phenylalanine), and wherein the pH of the formulation is 6.5.

[0043] In one embodiment, the formulation comprises 175 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof comprising a VH domain comprising the sequence set forth in SEQ ID NO:7, or a sequence at least 95% identical thereto, and a VL domain comprising the sequence set forth in SEQ ID NO:8; 250 mM arginine; 20 mM histidine buffer; 0.02% non-ionic surfactant; 75 mM phenylalanine, and wherein the pH of the formulation is 6.5.

[0044] In one embodiment, the formulation comprises 175 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof comprising a VH domain comprising the sequence set forth in SEQ ID NO:7, or a sequence at least 95% identical thereto, and a VL domain comprising the sequence set forth in SEQ ID NO:8; 250 mM Arg-HCl; 20 mM histidine buffer; 0.02% polysorbate 20; 75 mM phenylalanine, and wherein the pH of the formulation is 6.5.

[0045] In one embodiment, the formulation comprises 175 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof comprising a VH domain comprising the sequence set forth in SEQ ID NO:7, or a sequence at least 95% identical thereto, and a VL domain comprising the sequence set forth in SEQ ID NO:8; 250 mM Arg-HCl; 20 mM histidine buffer; 0.02% polysorbate 20; 75 mM phenylalanine, and wherein the pH of the formulation is 6.5. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0046] As used herein, "long term" refers to a period of at least 6 months (such as 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 months). In one embodiment, the storage of the formulations of the present disclosure is at least 12 months (such as 12, 18, and 24 months).

[0047] As used herein, "nonionic surfactant" refers to a surfactant having a covalently attached oxygen-containing hydrophilic group attached to a hydrophobic parent structure. Examples of nonionic surfactants include: fatty alcohol ethoxylates (such as narrow range ethoxylates, octaethylene glycol monododecyl ether and pentaethylene glycol monododecyl ether), alkylphenol ethoxylates (nonoxynols and Triton). X-100), ethoxylates such as fatty acid ethoxylates, ethoxylated amines and / or fatty acid amides (such as polyethoxylated tallow amine, cocamide monoethanolamine and cocamide diethanolamine), endblocked ethoxylates (such as poloxamers); fatty acid esters of polyhydroxy compounds; fatty acid esters of glycerol (such as glycerol monostearate and glycerol monolaurate); fatty acid esters of sorbitol (such as sorbitan monolaurate, sorbitan monostearate and sorbitan tristearate); Tweens (such as Tween 20, 40, 60 or 80); fatty acid esters of sucrose; alkyl polyglucosides (such as decyl glucoside, lauryl glucoside and octyl glucose); and polysorbates (such as polysorbate 20, 40, 60 or 80).

[0048] Thus, in one embodiment, the nonionic surfactant is selected from the group including ethoxylates; fatty acid esters of polyhydroxy compounds; fatty acid esters of glycerol; fatty acid esters of sorbitol; Tweens; fatty acid esters of sucrose; alkyl polyglucosides; and polysorbates.

[0049] As used herein, "parenteral formulation" refers to a formulation that is designed not to be delivered via the digestive tract. Typical parenteral delivery routes include injection (including bolus injection), implantation, or infusion. In one embodiment, the formulation is provided in the form of bolus delivery.

[0050] In one embodiment, the parenteral formulation is administered intravenously. In one embodiment, the parenteral formulation is administered subcutaneously.

[0051] As used herein, "injection" refers to administration of a liquid formulation into the body via a syringe or syringe pump. Injections include intravenous, subcutaneous, intratumoral, or intramuscular administration. Injections are generally over a short period of time, such as 5 minutes or less. However, injections may be administered slowly or continuously, for example, using a syringe pump. Injections generally involve administration of smaller volumes than infusions. In one embodiment, injections are administered as slow injections, for example, over a period of 1.5 to 30 minutes. As used herein, slow injections are manual injections using a syringe. In one embodiment, a dose of the formulation is less than 100 ml, for example 30 ml, such as administered by a syringe pump.

[0052] As used herein, "infusion" refers to the administration of a liquid by drip, infusion pump, or equivalent device. In one embodiment, the infusion is administered over a period ranging from 1 to 120 minutes (e.g., 1 to 5 minutes), such as about 1, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 65, 80, 85, 90, 95, 100, 105, 110, 115, or 120 minutes.

[0053] Anti-IL13R antibody Interleukin 13 receptor (IL-13R) as used herein is a type I cytokine receptor that binds interleukin 13. This receptor consists of two subunits, encoded by IL13Rα1 and IL4R, respectively. These two genes code for the proteins IL-13Rα1 and IL-4Rα, which form a dimer with IL-13 binding to the IL-13Rα1 chain, and IL-4Rα stabilizing this interaction. Due to the presence of the IL4R subunit, IL13R can also trigger IL-4 signaling. In either case, this occurs via activation of the Janus kinase (JAK) / signal transducer and activator of transcription (STAT) pathway, leading to phosphorylation of STAT6. Human IL-13Rα1 has the Uniprot number: P3597.

[0054] IL-13Rα2 (previously called IL-13R and IL-13Rα) is another receptor that can bind IL-13. However, in contrast to IL-13Rα1, this protein binds IL-13 with high affinity but does not bind IL-4. Human IL-13Rα2 has the Uniprot number: Q14627.

[0055] An anti-IL13R antibody herein refers to an antibody having specificity for IL13R, for example, IL13Rα1 or IL13Rα2.

[0056] In one embodiment, the anti-IL13R antibody of the present disclosure is specific for IL13Rα 1. In one embodiment, the anti-IL13R antibody binds to an epitope comprising the amino acid sequence FFYQ.

[0057] The anti-IL13R antibodies of the present disclosure may include complete antibody molecules having full-length heavy and light chains or binding fragments thereof, including, but not limited to, Fab, modified Fab, Fab', F(ab')2, Fv, single domain antibodies (such as VH, VL, VHH, V domains of IgNAR), scFv, bivalent, trivalent or tetravalent antibodies, Bis-scFv, diabodies, triabodies, tetrabodies, and epitope-binding fragments of any of the above (see, e.g., Holliger and Hudson, 2005, Nature Biotech. 23(9):1126-1136; Adair and Lawson, 2005, Drug Design Reviews - Online 2(3), 209-217).

[0058] Methods for making and producing these antibody fragments are known in the art (see, for example, Verma et al., 1998, Journal of Immunological Methods, 216, 165-181). Other antibody fragments for use in the present invention include the Fab and Fab' fragments described in WO2005 / 003169, WO2005 / 003170 and WO2005 / 003171. Other antibody fragments for use in the present invention include the Fab-Fv and Fab-dsFv fragments described in WO2010 / 035012 and antibody fragments comprising such fragments. Multivalent antibodies may comprise multiple specificities or may be monospecific (see, for example, WO92 / 22853 and WO05 / 113605).

[0059] Antibodies and fragments thereof for use in this disclosure may be from any species, including, for example, mouse, rat, shark, rabbit, pig, hamster, camel, llama, goat, or human. Chimeric antibodies have non-human variable regions and human constant regions.

[0060] The antibodies or binding fragments for use in the present invention may be from any class (e.g., IgG, IgE, IgM, IgD, or IgA) or subclass of immunoglobulin molecule. In one embodiment, the antibody used in this disclosure is an IgG4 or an IgG4 with a 241P mutation.

[0061] In one embodiment, the antibody or binding fragment used in the formulation of the present disclosure has an affinity of 5 nM or higher (higher affinities are lower numerical values), for example 500 pM, 250 pM or higher, particularly 125 pM or lower.

[0062] A sequence listing is provided as an annex hereto. GYSFTSYWIG (SEQ ID NO:1) VIYPGDSYTR (SEQ ID NO:2) MPNWGSLDH (SEQ ID NO:3) RASQSISSSYLA (SEQ ID NO:4) GASSRAT (SEQ ID NO:5) QQYAS (SEQ ID NO:6) SEQ ID NO:7 EVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGVIYPGDSYTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARMPNWGSLDHWGQGTLVTVSS SEQ ID NO:8 EIVLTQSPGTLSLSPGERATLSCRASQSISSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYASFGQGTKVEI* (*K is deleted in post-translational modification).

[0063] Anti-IL13R antibodies or binding fragments used in this disclosure include CDRH1 comprising the amino acid sequence set forth in SEQ ID NO:1, CDRH2 comprising the amino acid sequence set forth in SEQ ID NO:2, and CDRH3 comprising the amino acid sequence set forth in SEQ ID NO:3.

[0064] Anti-IL13R antibodies or binding fragments used in the present disclosure include CDRL1 comprising the amino acid sequence set forth in SEQ ID NO:4, CDRL2 comprising the amino acid sequence set forth in SEQ ID NO:5, and CDRL3 comprising the amino acid sequence set forth in SEQ ID NO:6.

[0065] In one embodiment, the VH sequence comprises SEQ ID NO: 7, or a sequence at least 95% identical thereto. In one embodiment, the VL sequence comprises SEQ ID NO: 8, or a sequence at least 95% identical thereto.

[0066] In one embodiment, the VH sequence comprises SEQ ID NO:7, or a sequence at least 95% identical thereto, and the VL sequence comprises SEQ ID NO:8, or a sequence at least 95% identical thereto.

[0067] In one embodiment, the VH sequence is SEQ ID NO:7 and the VL sequence is SEQ ID NO:8.

[0068] As used herein, a "variable region" refers to the region of an antibody chain that contains the CDRs and suitable frameworks.

[0069] In one embodiment, the heavy chain comprises a sequence independently selected from the group consisting of SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, and a sequence at least 95% identical to any of the above.

[0070] In one embodiment, the light chain comprises the amino acid sequence shown in SEQ ID NO:14, or a sequence at least 95% identical thereto.

[0071] In one embodiment, the heavy chain comprises SEQ ID NO:9, or a sequence at least 95% identical thereto, and the light chain comprises SEQ ID NO:14, or a sequence at least 95% identical thereto.

[0072] In one embodiment, the heavy chain comprises SEQ ID NO:10, or a sequence at least 95% identical thereto, and the light chain comprises SEQ ID NO:14, or a sequence at least 95% identical thereto.

[0073] In one embodiment, the heavy chain comprises SEQ ID NO:11, or a sequence at least 95% identical thereto, and the light chain comprises SEQ ID NO:14, or a sequence at least 95% identical thereto.

[0074] In one embodiment, the heavy chain comprises SEQ ID NO:12, or a sequence at least 95% identical thereto, and the light chain comprises SEQ ID NO:14, or a sequence at least 95% identical thereto.

[0075] In one embodiment, the heavy chain comprises SEQ ID NO:13, or a sequence at least 95% identical thereto, and the light chain comprises SEQ ID NO:14, or a sequence at least 95% identical thereto.

[0076] In one embodiment, the heavy chain is SEQ ID NO:9 and the light chain is SEQ ID NO:14.

[0077] In one embodiment, the heavy chain is SEQ ID NO:10 and the light chain is SEQ ID NO:14.

[0078] In one embodiment, the heavy chain is SEQ ID NO:11 and the light chain is SEQ ID NO:14.

[0079] In one embodiment, the heavy chain is SEQ ID NO:12 and the light chain is SEQ ID NO:14.

[0080] In one embodiment, the heavy chain is SEQ ID NO:13 and the light chain is SEQ ID NO:14.

[0081] As used herein, "derived from" refers to the fact that the sequence used, or a sequence closely similar to the sequence used, is obtained from the genetic material of a source, such as the light or heavy chain of an antibody.

[0082] As used herein, "at least 95% identical" is intended to refer to an amino acid sequence that is 95% or more identical to a reference sequence over its entire length, such as 96, 97, 98, or 99% identical. Software programs can be used to calculate percent identity.

[0083] In one embodiment, the antibody or binding fragment thereof used in the formulation of the present disclosure is humanized.

[0084] As used herein, "humanized" (which includes CDR-grafted antibodies) refers to a molecule having one or more complementarity determining regions (CDRs) from a non-human species and a framework region from a human immunoglobulin molecule (see, e.g., U.S. Pat. No. 5,585,089; WO 91 / 09967). It will be understood that it is only necessary to graft the specificity determining residues of the CDRs and not the entire CDR (see, e.g., Kashmiri et al., 2005, Methods, 36, 25-34). A humanized antibody may optionally further comprise one or more framework residues of the non-human species from which the CDRs are derived. For a review, see Vaughan et al., Nature Biotechnology, 16, 535-539, 1998.

[0085] When grafting CDRs or specificity determining residues, any suitable acceptor variable region framework sequence may be used, taking into account the class / type of the donor antibody from which the CDRs are derived, including mouse, primate and human framework regions. Examples of human frameworks that can be used in the present invention are KOL, NEWM, REI, EU, TUR, TEI, LAY and POM (Kabat et al., supra). For example, KOL and NEWM can be used for the heavy chain; REI can be used for the light chain; EU, LAY and POM can be used for both the heavy and light chains. Alternatively, human germline sequences can be used; these are available at http: / / vbase.mrc-cpe.cam.ac.uk / .

[0086] In the humanized antibodies used in the present invention, the acceptor heavy and light chains are not necessarily derived from the same antibody and may, if desired, comprise composite chains having framework regions derived from different chains.

[0087] The framework regions need not have exactly the same sequence as that of the acceptor antibody. For example, unusual residues may be exchanged for residues occurring more frequently in that class or type of acceptor chain. Alternatively, selected residues in the acceptor framework regions may be substituted such that they correspond to residues present at the same position in the donor antibody (see Reichmann et al., 1998, Nature, 332, 323-324). Such changes should be kept to the minimum necessary to restore the affinity of the donor antibody. Protocols for selecting residues in acceptor framework regions that may need to be changed are described in WO91 / 09967.

[0088] In one embodiment, the anti-IL13R antibodies of the disclosure are fully human, in particular one or more of the variable domains are fully human.

[0089] A fully human molecule is one in which the variable and constant regions (if present) of both the heavy and light chains are all of human origin or substantially identical to sequences of human origin, but not necessarily derived from the same antibody. Examples of fully human antibodies include, for example, antibodies made by the phage display method described above, as well as antibodies made by mice in which the mouse immunoglobulin variable and optionally constant region genes have been replaced by their human counterparts, such as those broadly described in European Patent Application No. 0546073B1, U.S. Patent No. 5,545,806, U.S. Patent No. 5,569,825, U.S. Patent No. 5,625,126, U.S. Patent No. 5,633,425, U.S. Patent No. 5,661,016, U.S. Patent No. 5,770,429, European Patent No. 0438474, and European Patent No. 0463151.

[0090] As used herein, "constant region" is intended to refer to the portion of the constant region located between two variable domains in a heavy chain, e.g., between noncognate variable domains. Thus, an anti-IL13R antibody of the disclosure may comprise one or more constant regions, such as a naturally occurring constant domain or a derivative of a naturally occurring domain.

[0091] As used herein, a "derivative of a naturally occurring domain" is intended to refer to one, two, three, four, or five amino acids in the naturally occurring sequence having been substituted or deleted to optimize the properties of the domain, such as by removing undesirable properties, while retaining the characteristics of the domain.

[0092] If necessary, the antibody for use in the present invention may be conjugated to one or more effector molecules. It will be understood that the effector molecule may comprise a single effector molecule, or two or more such molecules linked to form a single moiety that can be attached to the antibody of the present invention. If it is desired to obtain an antibody fragment linked to an effector molecule, this may be prepared by standard chemical or recombinant DNA methods in which the antibody fragment is linked to the effector molecule either directly or via a coupling agent. Techniques for conjugating such effector molecules to antibodies are known in the art (see Hellstrom et al., Controlled Drug Delivery, 2nd Edition, Robinson et al., eds., 1987, pp.623-53; Thorpe et al., 1982, Immunol.Rev., 62:119-58, and Dubowchik et al., 1999, Pharmacology and Therapeutics, 83, 67-123). Particular chemical means include, for example, those described in WO 93 / 06231, WO 92 / 22583, WO 89 / 00195, WO 89 / 01476, and WO 03031581. Alternatively, where the effector molecule is a protein or polypeptide, linkage may be achieved using recombinant DNA methods, for example as described in WO 86 / 01533 and EP 0 392 745.

[0093] The term "effector molecule" as used herein includes, for example, biologically active proteins, such as enzymes, other antibodies or antibody fragments, synthetic or natural polymers, nucleic acids and fragments thereof, such as DNA, RNA and fragments thereof, radionuclides, in particular radioactive iodides, radioisotopes, chelating metals, nanoparticles and reporter groups (such as fluorescent compounds or compounds which may be detected by NMR or ESR spectroscopy).

[0094] Other effector molecules can include detectable substances that are useful, for example, in diagnosis.Exemplary detectable substances include various enzymes, prosthetic groups, fluorescent substances, luminescent substances, bioluminescent substances, radionuclides, positron-emitting metals (for use in positron emission tomography), and non-radioactive paramagnetic metal ions.For metal ions that can be conjugated to antibodies for use as diagnostic methods, see U.S. Patent No. 4,741,900. Suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; suitable prosthetic groups include streptavidin, avidin, and biotin; suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride, and phycoerythrin; suitable luminescent materials include luminol; suitable bioluminescent materials include luciferase, luciferin, and aequorin; and suitable radionuclides include 125I, 131I, 111In, and 99Tc.

[0095] In another example, the effector molecule may extend the half-life of the antibody in vivo and / or reduce the immunogenicity of the antibody and / or facilitate delivery of the antibody across an epithelial barrier to the immune system. Examples of suitable effector molecules of this type include polymers, albumin, albumin-binding proteins, or albumin-binding compounds (such as those described in WO05 / 117984). When the effector molecule is a polymer, it may generally be a synthetic or natural polymer, such as an optionally substituted linear or branched polyalkylene, polyalkenylene, or polyoxyalkylene polymer, or a branched or unbranched polysaccharide, such as a homopolysaccharide or a heteropolysaccharide.

[0096] Particular optional substituents that may be present on the synthetic polymer include one or more hydroxy, methyl, or methoxy groups.

[0097] Specific examples of synthetic polymers include optionally substituted linear or branched polyethylene glycol, polypropylene glycol, polyvinyl alcohol or derivatives thereof, in particular optionally substituted polyethylene glycol (such as methoxypolyethylene glycol) or derivatives thereof.Specific natural polymers include lactose, amylose, dextran, glycogen or derivatives thereof.

[0098] As used herein, "derivative" is intended to include reactive derivatives, e.g., thiol-selective reactive groups such as maleimides. The reactive group may be linked to the polymer directly or via a linker segment. It will be understood that the residue of such a group will in some cases form part of the product as the linking group between the antibody fragment and the polymer.

[0099] Suitable polymers include polyalkylene polymers such as polyethylene glycol or in particular methoxypolyethylene glycol or derivatives thereof, and in particular those having a molecular weight in the range of about 15,000 Da to about 40,000 Da.

[0100] In one example, the antibody for use in the present invention is linked to a polyethylene glycol (PEG) moiety. In a specific example, the antibody is an antibody fragment, and the PEG molecule may be linked via any available amino acid side chain or functional group of the terminal amino acid located in the antibody fragment (e.g., any free amino, imino, thiol, hydroxyl, or carboxyl group). Such amino acids may occur naturally in the antibody fragment or may be engineered into the fragment using recombinant DNA methods (see, e.g., U.S. Pat. No. 5,219,996; U.S. Pat. No. 5,667,425; WO98 / 25971, WO2008 / 038024). In one example, the antibody molecule of the present invention is a modified Fab fragment, where the modification is the addition of one or more amino acids to the C-terminus of its heavy chain to allow for the attachment of an effector molecule. Suitably, the additional amino acids form a modified hinge region that includes one or more cysteine ​​residues to which the effector molecule may be attached. Multiple sites can be used to attach two or more PEG molecules.

[0101] In patients with cancer, such as breast cancer, cancer-related lymphedema (BCRL), the formulations of the present disclosure may prevent lymphedema-related effects such as fibrosis, hyperkeratosis, fibroadipose tissue deposition, fluid accumulation, limb swelling, loss of skin elasticity, and pain. By reducing excess volume, the formulations may improve lymphatic function and, for example, limb function.

[0102] The development of lymphedema following lymphatic injury is associated with tissue inflammation, infiltration of CD4 positive cells, and their differentiation into the type 2 helper T cell (Th2) phenotype. Th2 cells produce IL-4 and IL-13, which play key roles in the development of lymphedema-associated symptoms as well as other Th2-mediated diseases.

[0103] In one embodiment, the formulations described herein are administered in combination with another therapy.

[0104] As used herein, "in combination" is intended to include when an anti-IL13R antibody is administered prior to, simultaneously with, or following another treatment.

[0105] As used herein, a "therapeutic dose" refers to an amount of an anti-IL13R antibody (such as ebrasakimab) suitable for achieving an intended therapeutic effect, e.g., when used in a suitable treatment regimen that ameliorates a disease symptom or pathology, particularly without inducing dose-limiting side effects. A suitable therapeutic dose is generally a balance between therapeutic effect and tolerable toxicity, e.g., where side effects and toxicity are acceptable in view of the benefit achieved by the therapy.

[0106] In one embodiment, a formulation according to the present disclosure (including formulations comprising same) is administered once a month, for example in a treatment cycle or as a maintenance therapy.

[0107] In the context of this specification, "comprising" is interpreted as "including." Embodiments of the invention comprising a particular feature / element are also intended to extend to other embodiments "consisting of" or "consisting essentially of" the associated element / feature. Where technically appropriate, multiple embodiments of the invention may be combined.

[0108] Any amendments herein may be based on a priority document.

[0109] The "Background" section can be used as a basis for making corrections.

[0110] Technical references, such as patents and applications, are incorporated herein by reference.

[0111] Any embodiment specifically and explicitly recited herein may form the basis of a waiver either alone or in combination with one or more additional embodiments.

[0112] Implementations may be combined where technically appropriate.

[0113] The headings herein are used to divide the specification into sections and are not intended to be used to interpret the meaning of the disclosure provided herein.

[0114] The invention is further described, by way of illustration only, in the following examples. [Brief description of the drawings]

[0115] [Figure 1A] A schematic diagram of the Kinexus ultra+ rheometer used for viscosity measurements is shown. [Figure 1B] 1 shows a graph of the viscosity of the bulk drug product (BDP) of ebrasakimab. [Diagram 2] 1 is a series of graphs showing the viscosities of Formulations 1-15. [Diagram 3] 1 shows a graph comparing the viscosity of formulations containing different concentrations of Arg-HCl. [Figure 4] 1 shows a graph showing the viscosity of a formulation containing 150 mM Arg-HCl plus additional additives. [Diagram 5] 1 shows a graph comparing the viscosities of formulations 1-15. [Figure 6] 1 shows a comparison of viscosity measurements of ebrasakimab BDP obtained from stage 1 and stage 2 screening. [Figure 7] 1 shows a series of graphs comparing the viscosity of Formulations 3 and 6. [Figure 8] 1 shows a graph of repeated viscosity measurements of Formulation 3 to confirm reproducibility of viscosity measurements. [Figure 9] 1 shows a series of graphs showing the viscosity of formulations 16-30. [Figure 10] 1 shows a graph of the viscosity of Arg-Glu containing formulations versus Arg-HCl formulations. [Figure 11] 1 shows a graph showing the viscosity of a formulation containing 150 mM Arg-HCl plus additional additives. [Figure 12]1 shows a graph comparing the viscosity of formulations containing 150 mM Arg-HCl at different pH. [Figure 13] 1 shows a graph comparing the viscosity of 20 mM vs. 50 mM His-buffered formulations. [Figure 14] A graph comparing the viscosities of formulations 16 to 30 is shown. [Figure 15] 1 shows a series of graphs of viscosity measurements for formulations 31 and 32 in comparison with formulations 3 and 16. [Figure 16] A series of graphs of viscosity measurements for formulations 16-30 (formulation 40 = 200 mg / ml, all other formulations = 175 mg / ml) are shown. [Figure 17] A comparison of viscosity measurements for formulations 33-45 is shown. [Figure 18] 1 shows a graph of viscosity measurements for a 175 mg / ml ebrasakimab formulation. [Figure 19] 1 shows a graph of viscosity measurements for a 150 mg / ml ebrasakimab formulation. [Figure 20] 1 shows a graph of the results of a three-month stability study on viscosity. [Figure 21] 1 shows a graph of the results of a three-month stability study on osmolality. [Figure 22] A graph of the results of the 3-month stability study with respect to protein concentration (soloVPE) is shown for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 23] A graph of the results of the 3-month stability study with respect to pH is shown, for each formulation from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 24] 1 shows a table of the results of a three-month stability study based on visual inspection of the formulations. [Diagram 25]A graph of the results of the 3-month stability study for turbidity is shown, for each formulation from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 26] A table showing the results of the 3-month stability study on color is shown for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 27A] Graph showing the results of a 3-month stability study for subvisible particle (≧2 μm) content (MFI) using microflow imaging, for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 27B] A graph of the results of a 3-month stability study for subvisible particle (≧10 μm) content (MFI) for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 27C] Graph showing 3-month stability results for subvisible particle (≧25 μm) content (MFI) for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 28] A table showing the results of a 3-month stability study on protein mass recovery (HP-SEC) for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 29A] Graph of 3-month stability study results in terms of % aggregate content (HP-SEC) for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 29B]Graph of 3-month stability study results in terms of % monomer content (HP-SEC) for each formulation from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 29C] Graph of 3-month stability study results in terms of % fragment content (HP-SEC) for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 29D] 1 shows a summary table showing a comparison of % aggregate, monomer, and fragment content in a 100 mg / ml Ebrasakimab formulation (HP-SEC). [Figure 30A] Graph of 3-month stability study results for % non-reduced aggregate content (cGE) for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 30B] Graph of 3-month stability study results for % non-reduced monomer content (cGE) for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 30C] A graph of the results of a 3-month stability study on the % non-reduced intact protein content (cGE) for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 30D] 1 shows a comparative summary table of % intact protein content in 100 mg / ml Ebrasakimab formulations. [Figure 31A] Graph showing 3-month stability study results for reduced non-glycosylated heavy chain (NGHC) content % (cGE) for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 31B]Graph showing 3-month stability study results for % reduced heavy chain (HC) content (cGE) for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 31C] Graph of 3-month stability study results for reduced light chain (LC) content % (cGE) for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 31D] A graph of the results of a 3-month stability study on the % reduced LC+HC content (cGE) for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Figure 31E] Graph of 3-month stability study results for % content of reduced impurities (cGE) for each formulation, from left to right: t0; t1m, 05C; t1m, 25C; t1m, 40C; t3m, 05C; t3m, 25C; and t3m, 40C. [Fig. 31F] 1 shows a summary table comparing the % reduced LC+HC content and % NGHC content in the 100 mg / ml Ebrasakimab formulation (cGE). [Figure 32A] 1 shows a graph of the results of a 3-month stability study on % Acidic Peak Content (IEX). [Figure 32B] 1 shows a graph of the results of a 3-month stability study on % Native Peak Content (IEX). [Figure 32C] 1 shows a graph of the results of a 3-month stability study on Basic Peak Content % (IEX). [Fig. 32D] 1 shows a summary table comparing the % content of acidic peak, neutral peak, and basic peak in 100 mg / ml ASLAN004 formulation (IEX).

[0116] [Table 1]

[0117] Working Example Example 1: First Stage (1st) Screening: Formulations 1-15 Fifteen different formulations were initially created.

[0118] [Table 2]

[0119] The formulations were made as follows. (1) The ebrasakimab BDP sample was concentrated 2-fold to approximately 7.5 ml and diluted with 7.5 ml of formulation buffer. (2) The formulation was homogenized using a forced displacement pipette. (3) Steps 1 and 2 were repeated eight times. (4) After the eighth dilution step, each formulation was concentrated to obtain target concentrations of 150 mg / ml, 175 mg / ml, or 200 mg / ml. (5) Each formulation was sterile filtered. (6) Polysorbate-20 was added to achieve a target concentration of 0.02% (w / w). (7) Protein concentration (Solo-VPE), osmolality, and pH were measured for each formulation. (8) Vials were filled with formulation (1x1ml). (9) Any remaining samples were stored at 2-8 °C (CW18 viscosity / HP-SEC).

[0120] Viscosity Test The viscosity of each formulation was examined using a Kinexus ultra+ rheometer, see Figure 1A. Measurements were performed at 1000 rpm / sec at 20°C. The cone plates used were: 40mm, 1° angle, 24μm gap for samples <10cP(mPas) For samples >10cP (mPas), 20mm, 1° angle, 24μm gap

[0121] The average dynamic viscosity was calculated from measurements in the range of 540 to 600 seconds.

[0122] The ebrasakimab samples likely behave as non-Newtonian fluids, and the applied shear rate is constant. Therefore, in this disclosure, dynamic viscosity and shear viscosity are considered to be the same, and these terms are used synonymously in the examples.

[0123] FIG. 1B shows viscosity measurements for ebrasakimab BDP with added polysorbate 20.

[0124] The results of the viscosity measurements for formulations 1 to 15 are shown in FIG.

[0125] Table 2 below shows the formulations ranked by viscosity at different target concentration ranges.

[0126] [Table 3]

[0127] The above results show that the majority of the 150 mg / ml formulations and several of the 175 mg / ml formulations are close to the target viscosity of 20-25 cP.

[0128] Figure 3 shows the relationship between Arg-HCl concentration and viscosity. The results show that a minimum viscosity is reached at 150 mM Arg-HCl, and that higher Arg-HCl concentrations do not further reduce the viscosity. However, this was investigated further as follows.

[0129] Figure 4 shows the effect of different additives on viscosity. The results show that Arg-Glu reduces the viscosity similarly to Arg-HCl, and that the addition of additional additives to 150 mM Arg-HCl did not further reduce the viscosity. This was further investigated as follows.

[0130] FIG. 5 shows the correlation between viscosity and ASLAN004 concentration.

[0131] The results suggest that formulation 3 had the best overall viscosity across the three antibody concentrations.

[0132] The data herein is not a complete data set, but rather exemplary data to illustrate trends.

[0133] Example 2: Second Stage (2nd) Screening: Formulations 16-30 Using formulation 3 as a starting point, an additional 15 formulations were made in an attempt to develop a suitable 200 mg / ml ASLAN004 formulation. See Table 3 below.

[0134] [Table 4]

[0135] The above formulations were prepared as follows: (1) 10 ml of ebrasakimab BDP + 5 ml of formulation buffer was placed in an Amicon 15 unit (MWCO 100 kDa). (2) The formulation was concentrated 2-fold to approximately 7.5 ml and diluted with 7.5 ml of formulation buffer. (3) The formulation was homogenized using a forced displacement pipette. (4) Steps 2 and 3 were repeated four times. (5) After the fifth dilution step, each formulation was concentrated to obtain target concentrations of 150 mg / ml, 175 mg / ml, or 200 mg / ml. (6) Each formulation was sterile filtered (0.8 / 0.2 μm PES). (7) Polysorbate-20 was added to achieve a target concentration of 0.02% (w / w). (8) Protein concentration (Solo-VPE), osmolality, and pH were measured for each formulation. (9) Any remaining samples were stored at 2-8 °C (for viscosity determination).

[0136] Viscosity Test The viscosity of each formulation was examined using the same method as described in Example 1 above.

[0137] Figure 6 shows a comparison of viscosity measurements for ebrasakimab BDP from the first screen (Example 1) and the second screen (Example 2). As can be seen, the viscosity values ​​are very similar, suggesting that the ASLAN004 BDP used in both screens was comparable.

[0138] Figure 7 shows a comparison of formulations 3 and 16. Note that Figure 8 shows that the viscosity measurements were reproducible for formulation 3. These results demonstrated that the method is reliable and that the viscosity measurements are useful for evaluating formulations.

[0139] The results of the viscosity measurements for Preparations 16 to 30 are shown in FIG.

[0140] Table 4 below shows the formulations ranked by viscosity at different target concentration ranges.

[0141] [Table 5]

[0142] The results show that all 150 mg / ml formulations and several 175 mg / ml formulations were close to the target viscosity of 20 cP. Unexpectedly, many of the 200 mg / ml formulations had viscosity values ​​significantly lower than the predicted viscosity. Higher concentrations of Arg-HCl (such as 250 Mm) significantly reduced the viscosity in the 200 mg / ml formulations. 200 mg / ml formulations with higher concentrations of Arg-HCl (such as 250 Mm) in combination with salts such as CaCl2 (especially 50 Mm) and / or amino acids such as phenylalanine (especially 50-75 mM) can provide a viscosity of approximately 20 cP.

[0143] Figure 10 shows the viscosity of Arg-HCl and Arg-Glu formulations at various concentrations. The results suggest that there was a more significant decrease in viscosity for the Arg-HCl compared to the Arg-Glu formulation. In particular, increasing Arg-HCl from 175 mM to 250 mM appeared to significantly decrease the viscosity.

[0144] Figure 11 shows the effect of different additives on viscosity. The results show that the addition of phenylalanine and CaCl2 helps reduce the viscosity.

[0145] Figure 12 shows the effect of pH on viscosity for the 150 mM Arg-HCl formulation. The results show that increasing the pH from 6.5 to 7 or decreasing the pH from 6.5 to 6.0 did not improve the viscosity. Therefore, any pH between 6.0 and 7.0 is considered suitable.

[0146] Figure 13 shows the results of an experiment evaluating the effect of increasing concentrations of His buffer. The results suggest that increasing the histidine concentration from 20 mM to 50 mM for 175 mg / ml and 200 mg / ml ASLAN004 formulations reduces viscosity.

[0147] FIG. 14 shows the correlation between viscosity and SLAN004 concentration.

[0148] The results suggest that formulation 29 had the best overall viscosity across the three antibody concentrations.

[0149] Example 3: Third stage screening The objective of the third stage screening was to gain a better understanding of the viscosity differences observed between the first and second stage screening, as well as to confirm the viscosity range of the formulation for formulation stress testing.

[0150] Based on the first and second stage screening, formulations 3 / 16 and 29 had the best overall viscosity. Two additional formulations were prepared based on these formulations.

[0151] [Table 6]

[0152] The formulations were prepared as follows: (1) 20 ml of ASLAN004 BDP (100 mg / ml) was diluted to 63 ml (62 mg / ml). (2) Buffer exchange was performed with 8 dilution volumes (run time of approximately 24 hours). (3) The formulation was concentrated to 166 mg / ml by tangential flow filtration (TFF). (4) The formulation was concentrated to 208 mg / ml in an Amicon unit. (5) Each formulation was sterile filtered (0.8 / 0.2 μm PES). (6) Finally, polysorbate-20 was added. (7) For formulation 32, 100 mM Arg-HCl was added.

[0153] Viscosity Test The viscosity of each formulation was examined using the same method as described in Example 1 above.

[0154] The results are shown in FIG. 15 and Table 5 below.

[0155] [Table 7]

[0156] There was good agreement between the viscosities of formulations 29 and 32.

[0157] The results suggest that increasing the arginine concentration from 150 mM to 250 mM resulted in a significant and consistent improvement in the viscosity of 200 mg / ml ASLAN004.

[0158] Example 4: Fourth stage screening Using formulation 29 as a starting point, an additional 13 formulations were made to further improve the 175 mg / ml formulation, see Table 6 below.

[0159] [Table 8]

[0160] Formulations were prepared using the same method as described above for the second screen (Example 2). Formulations 41-45 were prepared 2 weeks after Formulations 33-40.

[0161] Viscosity Test The viscosity of each formulation was examined using the same method as described in Example 1 above.

[0162] The results of the viscosity measurements for preparations 33 to 45 are shown in FIG.

[0163] Table 7 below shows the formulations ranked by viscosity.

[0164] [Table 9]

[0165] Figure 17 shows the correlation between viscosity and Ebrasakimab concentration. The phenylalanine formulations showed a tendency to cluster at lower viscosity values, suggesting that the addition of phenylalanine to the formulation aids in viscosity reduction.

[0166] Example 5: Formulation Selection Viscosity results from the second through fourth screens were compared to determine the best formulation to proceed to stability testing.

[0167] Figure 18 shows a summary of viscosity measurements for the second, third, and fourth screens for 175 mg / ml Ebrasakimab formulations. The results suggest that the formulations tested in the fourth screen that contained Arg-HCl and phenylalanine had the lowest viscosities; namely, formulations 35, 38, 39, and 41.

[0168] Figure 19 shows a summary of the viscosity measurements of the second and third screens for the 150 mg / ml Ebrasakimab formulations. The results suggest that formulations 21 and 29 had the lowest viscosity.

[0169] Based on these results, the final list of formulations shown in Table 8 was selected.

[0170] [Table 10]

[0171] These formulations will be developed and advanced into stability / stress testing studies.

[0172] Example 6: Stability Study 3 Month Preliminary Results Six formulations in Table 8 were advanced into a stability testing study whereby the formulations were stored at 2-8° C., 25° C. or 40° C. Interim results over a three month period are shown in Figures 20-32.

[0173] Figure 20 shows the viscosity results for the six formulations. The results suggest that the viscosity increased slightly after three months. Notably, the viscosity was similar for samples stored at 5°C and 25°C, but a more significant increase in viscosity was observed for samples stored at 40°C.

[0174] Figure 21 shows the osmolality results for the six formulations. The differences observed were within the expected method variability.

[0175] Figure 22 shows the protein concentration results for the six formulations. There was no clear change observed in the 3 month samples compared to the day 0 samples.

[0176] Figure 23 shows the pH results for the six formulations. The differences observed were within the expected method variability.

[0177] Figure 24 shows the results of visual inspection of the six formulations. In general, the formulations had a milky appearance and particles were on the edge of visibility, with the exception of F49 at 40°C, suggesting that particle formation was minimal.

[0178] Figure 25 shows the turbidity results for the six formulations. The results indicate that the turbidity of the formulations was comparable between the day 0, 1 month, and 3 month samples. The lowest turbidity values ​​were observed for the samples stored at 25°C.

[0179] Figure 26 shows the color change results for six formulations. With the exception of F49 at 40°C, the remaining formulations were still in the brown spectrum (B5) after three months of storage. No differences in coloration were observed between samples stored at 2-8°C or 25°C for one and three months. Conversely, little change in coloration was observed after three months of storage at 40°C compared to one month at 40°C.

[0180] 27A-27C show the results of an experiment on the content of subvisible particles performed using microflow imaging (MFI). After 3 months, an increase in the content of particles with a size of ≧2 μm was observed in all samples, especially in samples stored at 40° C.

[0181] Figures 28 and 29A-29D show the results of experiments carried out using HP-SEC. Details of the equipment and parameters used are as follows: Instrument: Dionex Ultimate 3000; Column: Waters Xbridge protein BEH SEC, 7.8 x 300mm Detection: UV at 280 nm; Sample preparation: Diluted to 1 mg / ml (intermediate dilution, 50 mg / ml) in 20 mM histidine-HCl at pH 6.5; Measurements: n=1

[0182] FIG. 28 suggests that the % protein recovery for all formulations stored at 40° C. for 3 months was slightly lower compared to formulations stored at 5° C. or 25° C., however, did not appear to be significantly different between the formulations. Results in FIG. 29A-29C show a similar decrease in monomer content and increase in high molecular weight and fragment content across the six formulations. However, the loss of monomer was relatively low, with little loss after 3 months at 5° C., ≦1% loss after 3 months at 25° C., and ≦4% loss after 3 months at 40° C. A higher aggregate content was observed for F49 compared to the other formulations. FIG. 29D shows that the % intact protein concentration measured using SEC for formulation F49 was very similar to that of the 100 mg / ml ebrasakimab formulation.

[0183] Figures 30A-30D show the results of experiments performed using capillary gel electrophoresis (cGE) on non-reduced samples. Details of the equipment and parameters used are as follows: Instrument: SCIEX PA 800 plus with 30.2 cm exposed fused silica capillary. Sample preparation: Dilution of sample with 20 mM histidine in a two-step process (target concentration, 5 mg / ml). Measurement: Samples were split into two runs on different days (within precision, day-to-day reproducibility of duplicate measurements = 0.8%), n = 2.

[0184] Figures 30A-30C suggest that there was little loss of intact protein after 3 months at 5° C., mainly due to fragmentation, with less than 3% loss after 3 months at 25° C. and less than 15% loss after 3 months at 40° C. Figure 30D shows that the % intact protein concentration measured using cGE for formulation F49 was very similar to that of the 100 mg / ml ebrasakimab formulation.

[0185] 31A-31F show the results of experiments performed using capillary gel electrophoresis (cGE) on reduced samples. Figures 31A-31D show that there was no significant change in the relative content of light chain (LC). There was a slight decrease in the heavy chain (HC) content after 3 months at 25°C and 40°C. When both LC and HC content are taken into account simultaneously (Figure 31D), the results show that the LC+HC content was >99% after 3 months at 5°C and >98% after 3 months at 25°C. Figure 31E shows the impurity levels after 1 month and 3 months of storage. From these results, we believe that the decrease in the relative HC content after 3 months is partly due to deglycosylation of the reduced samples and also to an increase in the impurity levels over time. This trend was observed across the various formulations.

[0186] FIG. 31F shows that the LC+HC% and NGHC% measured for formulation F49 were very similar to those of the 100 mg / ml ebrasakimab formulation.

[0187] Figures 32A-32D show the results of an ion exchange chromatography (IEX) experiment. Details of the equipment and parameters used are as follows: Instrument: Vanquish UHPLC; Column: ProPac WCX-10, 4.0 x 250mm, 10μm; Detection: UV at 280nm; Sample preparation: Sample dilution to 1mg / ml (intermediate dilution, 50mg / ml) in 20mM MES, pH 7.0; Measurements: n=2

[0188] Figures 32A-32C show that there was similar neutral species loss, and similar acidic and basic species gain for all formulations over the 3 months of storage. There was little neutral species loss after 3 months at 5°C, less than 10% neutral species loss after 3 months at 25°C, and less than 45% neutral species loss after 3 months at 40°C. Figure 32D demonstrates that the acidic, neutral, and basic species content measured for formulation F49 was very similar to that of the 100 mg / ml ebrasakimab formulation.

[0189] In summary, the three-month interim stability data was very encouraging; the data suggested that all six formulations tested had good stability at the three-month time point. Based on this result, formulation F46 was selected as the first formulation to move forward.

Claims

1. A highly concentrated antibody formulation, 150-210 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof, for example 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205 or 210 mg / ml, particularly 150 mg / ml, 175 mg / ml or 200 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof; 170-250 mM arginine (such as Arg-HCl or Arg-Glu), for example 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245 or 250 mM, in particular 150 mM, 175 mM or 250 mM arginine; 20-50 mM histidine buffer, for example 20, 25, 30, 35, 40, 45 or 50 mM, such as 20 mM or 50 mM histidine buffer; 0.01-0.03% non-ionic surfactant, such as 0.02% w / w; Including; the pH of the formulation is in the range of 6.0 to 7.0, such as 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9 or 7.0, in particular pH 6.5; the anti-IL-13R antibody or antigen-binding fragment thereof VH CDR1 comprising SEQ ID NO: 1, VH CDR2 comprising SEQ ID NO: 2, VH CDR3 comprising SEQ ID NO: 3, VL CDR1 comprising SEQ ID NO: 4, a VL CDR2 comprising SEQ ID NO:5, and VL CDR3 comprising SEQ ID NO: 6, Including, formulation.

2. 2. The formulation of claim 1, wherein the anti-IL-13R antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 7 or a sequence at least 95% identical thereto.

3. 2. The formulation of claim 1, wherein the anti-IL-13R antibody comprises a VL domain comprising the amino acid sequence set forth in SEQ ID NO:8 or a sequence at least 95% identical thereto.

4. 10. The formulation of claim 1, comprising 150, 175, or 200 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof.

5. 10. The formulation of claim 1, comprising 175 mM, 200 mM, or 250 mM arginine, such as 250 mM arginine.

6. The formulation of claim 1, wherein the arginine is Arg-HCl or Arg-Glu, particularly Arg-HCl.

7. 10. The formulation of claim 1, comprising a 20 mM or 50 mM histidine buffer, such as a 20 mM histidine buffer.

8. 10. The formulation of claim 1, comprising 0.02% w / w of a non-ionic surfactant, such as 0.02% w / w of polysorbate 20.

9. 2. The formulation of claim 1, wherein the pH is 6.0, 6.5 or 7.0, such as 6.

5.

10. 2. The formulation of claim 1 further comprising a further amino acid, for example phenylalanine, 45 to 85 mM phenylalanine, such as 45, 50, 55, 60, 65, 70, 75 or 80 mM.

11. 11. The formulation of claim 10, comprising 50, 75 or 80 mM phenylalanine.

12. 10. The formulation of claim 1, further comprising CaCl2, e.g., 50 mM CaCl2.

13. 10. The formulation of claim 1, further comprising 50 to 200 mM of a sugar, such as sucrose.

14. 10. The formulation of claim 1, comprising 180 mM sugar, such as 180 mM sucrose.

15. 2. The formulation of claim 1, wherein the viscosity is less than 25 cP, in particular less than 20-25 cP, such as less than 20 cP.

16. 175 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof; 250 mM arginine, such as Arg-HCl; 20 mM histidine buffer; 0.02% non-ionic surfactant, such as 0.02% polysorbate 20; 75 mM phenylalanine, Including; the pH of the formulation is 6.5; The high-concentration antibody formulation according to claim 1 .

17. 175 mg / ml of an anti-IL-13R antibody or antigen-binding fragment thereof, comprising a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 7, or a sequence at least 95% identical thereto, and a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 8, or a sequence at least 95% identical thereto; 250 mM Arg-HCl; 20 mM histidine buffer; 0.02% polysorbate 20; 75 mM phenylalanine, Including; the pH of the formulation is 6.5; The high-concentration antibody formulation according to claim 1 .

18. A formulation according to any one of claims 1 to 17 for use in the treatment of an inflammatory disorder, for example atopic dermatitis, such as moderate to severe atopic dermatitis.

19. A formulation according to any one of claims 1 to 17 for use in therapy, in particular for the treatment of atopic dermatitis, such as moderate to severe atopic dermatitis.

20. 20. Use of a formulation according to any one of claims 1 to 17 in the manufacture of a medicament for use in the treatment of atopic dermatitis, such as moderate to severe atopic dermatitis.

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