Lyophilized formulation comprising GM-CSF neutralizing compound

Abstract

The present invention relates to aqueous formulations comprising at least 20 mg/ml of a compound neutralizing GM-CSF, a lyoprotectant and an amino acid and/or a buffer. The ingredients of the formulation preferably provide stability to the compound neutralizing GM-CSF in view of lyophilization, storage and reconstruction. In a preferred aspect, the formulation, e.g. after reconstruction, is for use in therapy, preferably for use in the treatment of inflammatory and autoimmune disorders, preferably including allergic and psoriatic disorders, as well as arthritic and asthmatic disorders. Furthermore, a kit comprising the formulation of the invention is provided.

Claims

1. A composition comprising (a) an antibody or a functional fragment thereof binding to GM-CSF or to the GM-CSF receptor and present in a concentration from about 50 mg/ml to about 150 mg/ml; (b) one or more lyo-protectants selected from one or more of sucrose, trehalose and mannitol and present in a concentration from about 50 mM to about 300 mM; (c) an amino acid selected from either or both histidine, and arginine, which amino acid is present in a concentration from about 10 mM to about 30 mM, and/or a buffer selected from one or more of histidine, arginine and citrate buffer, which buffer is present in a concentration from about 10 mM to about 30 mM; and (d) a surfactant in an amount from about 0.001% to about 1% (w/w), wherein the pH is between about 5 and, about 7, and wherein the composition does not form more than 8% aggregates relative to the amount of the antibody or functional fragment thereof at the beginning of storage, within a shelf life of at least 2 years at refrigerated or ambient conditions after lyophilization.

2. The composition according claim 1, wherein the surfactant is polysorbate 80.

3. A method of producing a lyophilized composition of the composition according to claim 1, in a lyophilization process comprising at least one of a freezing step, an annealing step and a drying step.

4. A lyophilized composition obtained by the method according to claim 3.

5. The composition according to claim 1, wherein the antibody or the functional fragment thereof is a human monoclonal antibody or a functional fragment thereof.

6. The composition according to claim 1, wherein the antibody is an IgG, an IgG1 or an IgG4 antibody.

7. The composition according to claim 1, wherein said antibody or functional fragment thereof comprises in its heavy chain variable region a CDR3 comprising an amino acid sequence chosen from the group consisting of those set out in any of the SEQ ID NOs: 1-13 and 56.

8. The composition according to claim 7, wherein any of said heavy chain variable region CDR3 sequences exists together with the heavy chain variable region CDR1 comprising the amino acid sequence set out in SEQ ID NO: 14 and heavy chain variable region CDR2 comprising the amino acid sequence set out in SEQ ID NO: 15.

9. The composition according to claim 1, wherein said antibody or functional fragment thereof comprises in its light chain variable region a CDR1 comprising the amino acid sequence set out in SEQ ID NO: 16, a CDR2 comprising the amino acid sequence set out in SEQ ID NO: 17, and a CDR3 comprising the amino acid sequence set out in SEQ ID NO: 18.

10. The composition according to claim 1, wherein said antibody or functional fragment thereof comprises in its light chain variable region an amino acid sequence as set out in any of SEQ ID NOs. 19, 54 and 55.

11. The composition according to claim 1, wherein said antibody or functional fragment thereof comprises a light chain amino acid sequence as set out in SEQ ID NO: 34 and a heavy chain amino acid sequence chosen from the group consisting of those as set out in any of SEQ ID NOs: 35-48.

12. A composition which comprises i) about 75 mg/ml of an antibody or a functional fragment thereof binding to GM-CSF or to the GM-CSF receptor, ii) about 62.5 mg/ml sucrose, iii) about 15.0 mM L-Histidine, iv) about 0.10 mg/ml polysorbate 80 and v) has a pH of about 5.8.

13. The A composition reconstituted after lyophilization-which comprises i) about 130 mg/ml of an antibody or a functional fragment thereof binding to GM-CSF or to the GM-CSF receptor which comprises, ii) about 108 mg/ml sucrose, iii) about 26.0 mM L-Histidine, iv) about 0.17 mg/ml polysorbate 80 and v) has a pH of about 5.8.

14. The composition according to claim 1 or claim 13, which is for intravenous and/or subcutaneous administration.

15. The composition according to any of claim 1, claim 4 or claim 13 for use in therapy.

16. The composition of claim 15 for use in the treatment of inflammatory and autoimmune disorders, preferably including allergic and psoriatic disorders, as well as arthritic and asthmatic disorders.

17. A kit comprising the composition according to any of claim 1, claim 4 or claim 13, wherein said composition may be is liquid, lyophilized or reconstituted.

18. The composition according to claim 1, wherein said antibody or functional fragment thereof comprises in its heavy chain variable region an amino acid sequence as set out in any of SEQ ID NOs: 20-33, 52 and 53.

Description

EXAMPLE 1

(1) Trehalose-Citrate Polysorbate Formulation

(2) TABLE-US-00007 ANTI-GM-CSF ANTIBODY 100 mg/mL Trehalose 100 mg/mL Citric acid 20 mM Polysorbate 80 0.2 mg/mL NaOH(2M) q.s. pH 5.8

(3) After six weeks storage at 40° C. the samples with a fill weight of 1.6 g were reconstituted with 1.0 mL of purified water, resulting in 120 mg/mL of anti-GM-CSF antibody, showed less than 3% aggregation.

EXAMPLE 2

(4) Sucrose-Mannitol-Histidine Formulation

(5) TABLE-US-00008 ANTI-GM-CSF ANTIBODY 90 mg/mL Sucrose 25 mg/mL Mannitol 12.5 mg/mL L-histidine 15 mM HCl (2M) q.s. pH 5.8

(6) After six weeks storage at 40° C. the samples with a fill weight of 2.0 g were reconstituted with 1.0 mL of purified water, resulting in 140 mg/mL of anti-GM-CSF antibody showed less than 3% aggregation.

EXAMPLE 3

(7) Sucrose-Histidine-Polysorbate Formulation

(8) TABLE-US-00009 ANTI-GM-CSF ANTIBODY antibody 75 mg/mL Sucrose 62.5 mg/mL L-Histidine 15 mM Polysorbate 80 0.1 mg/mL HCl (2M) q.s. pH 5.8

(9) After six weeks storage at 40° C. the samples with a fill weight of 2.5 g were reconstituted with 1.2 mL of purified water, resulting in 130 mg/mL of anti-GM-CSF antibody showed less than 2% aggregation.

(10) In-Use Stability

(11) The in-use stability of the reconstituted solution in the 10R glass vials was determined by size exclusion chromatography for the parameter aggregation and surface plasmon resonance for the parameter active concentration over 24 hours at ambient conditions. The active concentration of the reconstituted solution did not change over 24 hours at ambient conditions and the increase in aggregation was minor (from 0.64% to 0.76%).

(12) Short Term Stability

(13) After reconstitution with water for injection the samples are analysed. The key quality attribute is the degree of aggregation. This is determined by Size Exclusion Chromatography.

(14) TABLE-US-00010 Temperature Storage time [° C.] relative Aggregates Protein content intervall: humidity (r.h.) [%] [mg/ml] T.sub.0 5 0.5 130 +− 10% 1 months 5 0.5 130 +− 10% 3 months 5 0.6 130 +− 10% 6 months 5 0.6 130 +− 10% T.sub.0 25; 60% r.h. 0.5 130 +− 10% 1 months 25; 60% r.h. 0.6 130 +− 10% 3 months 25; 60% r.h. 0.9 130 +− 10% 6 months 25; 60% r.h. 1.1 130 +− 10% T.sub.0 40; 75% r.h. 0.5 130 +− 10% 1 months 40; 75% r.h. 1.1 130 +− 10% 2 months 40; 75% r.h. 1.6 130 +− 10% 3 months 40; 75% r.h. 1.9 130 +− 10%

(15) After 3 months at all storage conditions the aggregates are below 2%.

(16) Long Term Stability:

(17) The samples have been stored at 5° C.+−3° C. for the indicated time intervalls in the 10R vials. After reconstitution with water for injection the samples are analysed. The key quality attribute is the degree of aggregation. This is determined by Size Exclusion Chromatography.

(18) TABLE-US-00011 Storage time Aggregates Fragments Protein content intervall: [%] [%] [mg/ml] 0 0.5 <1.0 130 +− 10% 3 months 0.6 <1.0 130 +− 10% 6 months 0.6 <1.0 130 +− 10% 9 months 0.7 <1.0 130 +− 10% 12 months 0.7 <1.0 130 +− 10% 18 months 0.7 <1.0 130 +− 10%
Viscosity Evaluation:

(19) Viscosity is determined with a rheometer. A rheometer is used for those fluids which cannot be defined by a single value of viscosity and therefore require more parameters to be set and measured than is the case for a viscometer.

(20) For some fluids, viscosity is a constant over a wide range of shear rates (Newtonian fluids). The fluids without a constant viscosity (non-Newtonian fluids) cannot be described by a single number. Non-Newtonian fluids exhibit a variety of different correlations between shear stress and shear rate. Therefore, the viscosity is dependent on the temperature as well as the shear rate for non-Newtonian fluids.

(21) The viscosity is given in milli Pascal*seconds (mPa*s) at a given temperature and a given shear rate. An acceptable viscosity for the formulation to be lyophilized and for the reconstructed solution for administration is only slightly elevated in comparison to water. Preferably it is below 20 mPa*s at a shear rate between about 50 and about 1000 1/s at a temperature of 20° C. More preferably it is below 15 mPa*s at a temperature of about 20° C. and a shear rate of between 50 and 1000 1/s.

(22) The viscosity of the reconstituted ANTI-GM-CSF ANTIBODY formulation with 130+−10 mg/ml of antibody is below 15 mPa*s at a shear rate γ between 50 and 1000 1/s at a temperature of 20° C.

(23) Liquid Formulation:

(24) Paragraph for the Description:

(25) The viscosity of the formulation preferably is below 20 mPa*s at a shear rate between about 50 and about 1000γ at a temperature of 20° C. More preferably it is between about 2 and 10 mPa*s.

(26) The viscosity of the HCL formulation with 150 mg/ml of antibody is below 12 mPas*s at a shear rate between about 50 and about 1000 1/s at a temperature of 20° C.

(27) The viscosity of the HCL formulation with 150 mg/ml of antibody is below 20 mPas*s at a shear rate between about 50 and about 1000 1/s at a temperature of 5° C.