WO2019240690A1

WO2019240690A1 - Aqueous pharmaceutical composition of etanercept

Info

Publication number: WO2019240690A1
Application number: PCT/TR2018/050302
Authority: WO
Inventors: Guniz CAKAR; Ozgur KARBAN; Claudio PAVIA; Gabriel Ignacio Amadeo; Laura Valeria MAURO; Romina Paola ZUQUELI; Maximo BARRERAS; Maria Belén BOSCO; Eduardo ORTI; Ángela Guillermina FORNO
Original assignee: Centurion Ilac San. Ve Tic. A.S.
Priority date: 2018-06-11
Filing date: 2018-06-12
Publication date: 2019-12-19
Also published as: TR201808283A2

Abstract

The present invention relates to an aqueous pharmaceutical composition containing anti-tumor necrosis factor (TNF) agent which is administered by subcutaneous injection. The inventive pharmaceutical composition further comprises one or more excipients such as a stabilizer, an osmotic agent, an isotonic agent and buffer solution.

Description

AQUEOUS PHARMACEUTICAL COMPOSITION OF ETANERCEPT

T echnical Field

The present invention relates to an aqueous pharmaceutical composition containing anti -tumor necrosis factor (TNF) agent which is administered by subcutaneous injection.

Background of the Invention

Etanercept is a biological drug that treats autoimmune diseases by inhibiting tumor necrosis factor (TNF); a soluble inflammatory cytokine. It is a macromolecule with approximately 150 kDa molecular weight. It was firstly marketed by Amgen in 2002 under the trade name of E nbrel^÷ .

Etanercept is indicated for the treatment of rheumatoid arthritis, juvenile rheumatoid arthritis, soriatic arthritis, plaque psoriasis and ankylosing spondylitis.

Since Etanercept is a dimeric fusion protein, chemical and physical denaturation can be easily occurred and it causes loss of Etanercept^“s physiological activities.

The European patent document no. EP1478394 discloses an aqueous pharmaceutical composition suitable for long-term storage of polypeptides containing an Fc domain of an immunoglobulin. The said pharmaceutical composition essentially contains L-arginineas an aggregation inhibitor.

The European patent document no. EP2714009 discloses a stable liquid formulation of Etanercept (recombinant p75 sTNFR:Fc fusion protein), and more particularly, to a liquid formulation comprising one or more stabilizers selected from the group consisting of methionine, lysine, histidine, and pharmaceutically acceptable salts thereof in an amount sufficient to reduce by-product formation of Etanercept during storage. T he liquid formulation according to the said patent document effectively reduces production of Etanercept by-products and stably maintains its pharmaceutical efficacies for long-term storage. Therefore, the reconstitution procedure is not required before admi nistration, and the sterile formulation can be administered to patients to ensure patient safety. Thus, it can be appl i ed to the f i el ds i n need of E tanercept treatment.

A nother E uropean patent document no. E P2919801 discloses the stable pharmaceutical compositions comprising tumor necrosis factor receptor Fc fusion protein (T NF R: Fc). More particularly, the patent document relates to the stable pharmaceutical compositions comprising tumor necrosis factor receptor Fc fusion protein (T NFR:Fc), phosphate- citrate buffer. It also relates to the methods of manufacturing the composition, method of administration and kits containing the same.

Summary of the Invention

A n objective of the present invention is to realize an aqueous pharmaceutical composition stabilized without L-Arg in order to offer the alternative formulations of Etanercept have lower costs.

A n objective of the present invention is to realize an aqueous pharmaceutical composition which is stable and having less impurities derived from the product.

A nother objective of the present invention is to realize an aqueous pharmaceutical composition, essentially free of arginine, comprising stabilizer in order to prevent protei n aggregati on and to mai ntai n the activity of E tanercept for a I ong ti me. A further objective of the present invention is to realize an aqueous pharmaceutical composition, essentially free of arginine, comprising stabilizer in order to i ncrease storage stability of Etanercept.

Detailed Description of the Invention

T he inventive aqueous pharmaceutical composition essentially comprises:

-at least one therapeutically active ingredient which is an anti-tumor necrosis factor (T NF) agent,

-at least one stabilizer which prevents protein aggregation and provides stabi I ity of protei n duri ng i n the bul k sol uti on, duri ng processi ng and duri ng the shelf life thereof,

-at least one pharmaceutically acceptable salt, and buffer solution.

In the aqueous pharmaceutical composition according to the present invention, active i ngredi ent i s E tanercept. In the preferred embodi ment, E tanercept whi ch used as active i ngredi ent may be produced by recombi nant D NA technol ogy i n a C hi nese hamster ovary (C HO) mammalian cell expression system.

T he Etanercept of the present invention is a biological inflammation modulator that functions as a competitive inhibitor of T NF- , binding to cell surface T NF- receptor, to inhibit T NF- mediated immune responses, and is used to treat rheumatoid, juvenile rheumatoid and psoriatic arthritis, plaque psoriasis and ankylosing spondylitis.

Specifically, in one preferred embodiment of the present invention, Etanercept is present in a concentration of 1 to 500 mg/ml, preferably 50 mg/ml.

According to the present invention, in order to stabilize the protein in solution at least one stabilizer increasing its solubility and reducing their aggregation is used. Since Etanercept is a dimeric fusion protein, chemical and physical denaturation can be easily occurred and it causes loss of Etanercept^“s physiological activities. T he aqueous pharmaceutical composition according to the present invention comprises one or more stabilizers selected from the group consisting of L- T hreonine, Taurine, L -V aline, N -acetyl- L -cysteine and PE G 400.

L-Threonine is a polar amino acid, and it has been suggested that can prevent aggregation of monoclonal antibody if they are subjected to acidic conditions. T herefore, in one embodiment of the present invention, L-Threonine is used as stabilizer in Etanercept formulation. In the said embodiment of the invention, L- T hreonine is present about 10 mM to 100 mM, preferably 50 nM .

T aurine is a natural sulfonic acid and it has been demonstrated that its amine group interacts with proteins conferring them a stabilizing effect. A lso, it is reported that decreases the viscosity of proteins in solution. Therefore, in another embodiment of the present invention, T aurine is used as stabilizer in Etanercept formulation. In the sai d embodi merit of the i nventi on, T auri ne i s present about 10 mM to 100 mM , preferably 50 mM .

In other embodiment of the present invention, the formulation contains about 1 to 50 mM L -V aline and/or PE G 400 as stabilizer. Preferably 14 to 30 mM L -V ali ne and/or PE G 400 is present in the composition. More preferably 25 mM L -V aline and/or PE G 400 is present in the preferred composition.

In another embodiment of the present invention, the formulation contains about 1 to 50 mM N-acetyl-L -cysteine and/or PE G 400 as stabilizer. Preferably 14 to 30 mM N-acetyl-L -cysteine and/or PE G 400 is present in the composition. More preferably 25 mM N-acetyl-L -cysteine and/or PE G 400 is present in the preferred composition.

Additionally, in the inventive aqueous pharmaceutical composition, in order to stabilize the protein in the bulk solution during processing and during the shelf life thereof comprises sucrose as stabilizer. Sucrose is also used as osmotic agent. T he aqueous pharmaceutical composition according to the present invention comprises sucrose in a concentration of 0.5 to 25 mg/ml, preferably 10 mg/ml.

In the aqueous pharmaceutical composition according to the present invention comprises an aqueous buffer, wherein the aqueous buffer is sodium and/or potassium phosphate buffer. T he aqueous buffer maintains the ionic force and the pH of the solution in which the protein is soluble and stable. Sodium phosphate monobasic monohydrate and sodium phosphate dibasic anhydrous forms the said aqueous buffer solution. In one preferred embodiment of the present invention; buffer solution contains sodium phosphate monobasic monohydrate is present in a concentration of 0.1 to 10.0 mg/ml and sodium phosphate dibasic anhydrous is present in a concentration of 0.01 to 5.0 mg/ml, preferably 2.6 mg/ml and 0.9 mg/ml respectively.

T he inventive aqueous pharmaceutical composition additionally contains at least one pharmaceutically acceptable salt in order to provide the ionic strength and creates a suitable chemical environment for the protein conservation. T he salt can be selected from the group consisting of sodium chloride and potassi um chloride. In one preferred embodiment of the present invention, sodium chloride is present in a concentration of 0.01 to 20 mg/ml, preferably 5.80 mg/ml (100mM).

Finally, the inventive aqueous pharmaceutical composition has water for injection as a dil uent.

According to the present invention, aqueous pharmaceutical composition has pH of 4.00 to 8.00, preferably 6 to 6.60.

T he inventive aqueous pharmaceutical composition is administered by subcutaneous injection. Additionally, the aqueous pharmaceutical composition according to the present invention is in a pre filled syringe.

Hereafter, the present invention will be described in details with reference to Examples. However, these Examples are for illustrative purposes only, and the invention cannot be li mited by these Examples.

E xample 1

With 500 ml of Etanercept solution about 1000 syringes of Etanercept 25 mg/0.5 ml are filled, without taking into account losses or discards. With 1000 ml of Etanercept solution about 1000 syringes of Etanercept 50 mg/1.0 ml are filled, without taking into account losses or discards. Sucrose, Sodium chloride, L- T hreonine, Sodium phosphate monobasic monohydrate, Sodium phosphate dibasic anhydrous are dissolved by gentle sti rring in a glass container, sterilized and despy rogeni zed, containing a volume of the 80 % of the final volume of water for injection. Etanercept is added to this solution by gentle stirring, avoidi ng foam formati on. D i I ute to fi nal vol ume with water for i nj ecti on, previ ous pH adj ustment to 6.00- 6.60. Lastly, Etanercept solution is subjected to sterile filtration.

T able 1 : T he composition of Example 1 (E 1 )

T able 2: T he composition of Example 1.1 (E 1.1 )

E xample 2

With 1000 ml of Etanercept solution about 1000 syringes of Etanercept 50 mg/1.0 ml are filled, without taking into account losses or discards. Sucrose, Sodium chloride, Taurine, Sodium phosphate monobasic monohydrate, Sodium phosphate di basi c anhydrous are dissolved by gentl e sti rri ng i n a glass contai ner, steri I i zed and despy rogeni z ed, contai ni ng a vol ume of the 80 % of the f i nal vol ume of water for injection. Etanercept is added to this solution by gentle stirring, avoidi ng foam formati on. D i I ute to fi nal vol ume with water for i nj ecti on, previ ous pH adj ustment to 6.00- 6.60. Lastly, Etanercept solution is subjected to sterile filtration.

T able 3: T he composition of Example 2 (E2)

T able 4: T he composition of Example 2.1 (E2.1 )

In the formulation development, excipients and conditions were tested to study how they coul d affect the behavi our of the protei n i n the bul k sol uti on and i n the f i ni shed product. F i nal ly the most suitabl e condi ti ons to keep i n ti me the protei n i n sol uti on no denatured and therefore able to perform their biological activity were determi ned. E xperimental R esults

In order to prove that the stability of the developed composition according to the present invention, some analytical tests were performed. Protein content, related proteins and high molecular weight impurities of Example 1 , 1.1 , 2, 2.1 and the f ormul ati on with L - A rgi ni ne ( I i ke E nbrel ÷ ) i nstead of L -T hreoni ne or T auri ne was analysed at stability condition at 2-8 i§.

T able 5: Stability Results of Protein content

C : A n aqueous pharmaceutical composition with 50 mM L-A rgi nine

T able 6: Stability Results of Related Proteins

5 T able 7: Stability Results of H igh molecular weight impurities

T he data presented above in Tables 5, 6, and 7 demonstrate that relative 10 i mprovements for alternative f ormul ati ons of E tanercept formul ati on contai ni ng L -

A rg as the stabilizer. A n aqueous pharmaceutical composition of Etanercept with 50 mM L -A rginine (C), an aqueous pharmaceutical composition with 25 and 50 mM L -T hreonine (E 1 and E 1.1 respectively) and an aqueous pharmaceutical composition with 25 and 50 mM Taurine (E2 and E 2.1 respectively) were analysed at stability condition at 2-8 i§.

T abl e 6 demonstrate that the f ormul ati ons contai ni ng T auri ne are more effi c i ent for preventing the formation of truncated forms and the formulations containing both T aurine and L -Threonine are more efficient for preventing formation of misfolded forms (12 months) as determi ned by H PL C-HIC (Hydrophobic Interaction C hromatography) .

According to the data presented in Table 7, the formulations containing both Taurine and L -T hreonine are more efficient for preventing formation of high molecular weight impurities as determined by H PLC-SE C (Size Exclusion C hromatography) .

W i thi n these basi c concepts, i t i s possi bl e to devel op a great vari ety of embodi ments of the inventive ^'Aqueous Pharmaceutical Composition of Etanercept it cannot be limited to the examples disclosed herein and it is essentially according to the claims.

Claims

C L AIM S

1. A n aqueous pharmaceutical composition essentially comprising:

-at least one pharmaceutically acceptable salt, and buffer solution.

2. T he aqueous pharmaceutical composition according to C laim 1 , wherein the active ingredient is Etanercept.

3. T he aqueous pharmaceutical composition according to C laim 2, wherein Etanercept is produced by recombinant D NA technology in a Chinese hamster ovary (C HO) mammalian cell expression system.

4. T he aqueous pharmaceuti cal composition accordi ng to C lai m 2 or 3, wherei n the formulation is comprising 1 to 500 mg/ml Etanercept.

5. T he aqueous pharmaceutical composition according to any of Claim 2 to 4, wherein the formulation is comprising 50 mg/ml Etanercept.

6. T he aqueous pharmaceutical composition according to C laiml , wherein the stabilizer used in the formulation is selected from the group consisting of L- T hreonine, Taurine, L -V aline, N -acetyl- L -cysteine and PE G 400.

7. T he aqueous pharmaceutical composition according to C laim 6, wherein the formulation is comprising 10 mM to 100 mM L-T hreonine.

8. T he aqueous pharmaceutical composition according to C laim 7, wherein the formulation is comprising 50 mM L-Threonine.

9. T he aqueous pharmaceutical composition according to C laim 6, wherein the formulation is comprising 10 mM to 100 mM Taurine.

10. T he aqueous pharmaceutical composition according to C laim 9, wherein the formulation is comprising 50 mM Taurine.

1 1. T he aqueous pharmaceutical composition according to C laim 6, wherein the formulation is comprising 1 mM to 50 mM L-V aline.

12. T he aqueous pharmaceutical composition accordi ng to C laim 1 1 , wherein the formulation is comprising 14 mM to 30 mM L-V aline.

13. T he aqueous pharmaceutical composition accordi ng to C laim 1 1 , wherein the formulation is comprising 25 mM L-V aline.

14. T he aqueous pharmaceutical composition according to C laim 6, wherein the formulation is comprising 1 mM to 50 mM N-acetyl-L -cysteine.

15. T he aqueous pharmaceutical composition accordi ng to C laim 14, wherein the formulation is comprising 14 mM to 30 mM N-acetyl-L -cysteine.

16. T he aqueous pharmaceuti cal composi ti on accordi ng to C I ai m 14 or 15, wherei n the formulation is comprising 25 mM N-acetyl-L-cysteine.

17. T he aqueous pharmaceutical composition according to C laim 6, wherein the formulation is comprising 1 mM to 50 mM PE G 400.

18. T he aqueous pharmaceutical composition accordi ng to C laim 17, wherein the formulation is comprising 14 mM to 30 mM PE G 400.

19. T he aqueous pharmaceuti cal composi ti on accordi ng to C I ai m 17 or 18, wherei n the formulation is comprising 25 mM PE G 400.

20. T he aqueous pharmaceuti cal compositi on accordi ng to the any of the precedi ng claims, wherein the formulation is further comprising sucrose as a stabilizer and osmotic agent.

21. T he aqueous pharmaceutical composition accordi ng to C laim 20, wherein the formulation is comprising sucrose in a concentration of 0.5 to 25 mg/ml.

22. T he aqueous pharmaceutical composition according to the Claims 20 or 21 , wherein the formulation is comprising sucrose in a concentration of 10 mg/ml.

23. T he aqueous pharmaceuti cal compositi on accordi ng to the any of the precedi ng claims, wherein the aqueous buffer used in the formulation is sodium and/or potassium phosphate buffer.

24. T he aqueous pharmaceutical composition accordi ng to C laim 23, wherein the aqueous buffer contains sodium phosphate monobasic monohydrate and sodium phosphate dibasic anhydrous.

25. T he aqueous pharmaceuti cal compositi on accordi ng to C I ai m 23 or 24, wherei n the formulation is comprising 0.1 to 10.0 mg/ml sodium phosphate monobasic monohydrate.

26. T he aqueous pharmaceuti cal compositi on accordi ng to C I ai m 23 to 25, wherei n the formulation is comprising 2.6 mg/ml sodium phosphate monobasic monohydrate.

27. T he aqueous pharmaceuti cal composi ti on accordi ng to C I ai m 23 or 24, wherei n the formulation is comprising 0.01 to 5.0 mg/ml sodium phosphate dibasic anhydrous.

28. T he aqueous pharmaceutical composition accordi ng to C laim 27, wherein the formulation is comprising 0.9 mg/ml sodium phosphate dibasic anhydrous.

29. T he aqueous pharmaceuti cal compositi on accordi ng to the any of the precedi ng claims, wherein pharmaceutically acceptable salt used in the formulation is selected from the group consisting of sodium chloride and potassium chloride.

30. T he aqueous pharmaceutical composition accordi ng to C laim 29, wherein the formulation is comprising 0.01 to 20 mg/ml sodium chloride.

31. T he aqueous pharmaceutical composition according to the C laim 29 or 30, wherein the formulation is comprising 5.80 mg/ml sodium chloride.

32. T he aqueous pharmaceuti cal compositi on accordi ng to the any of the precedi ng claims, wherein the diluent used in the formulation is water for injection.

33. T he aqueous pharmaceuti cal compositi on accordi ng to the any of the precedi ng claims, wherein pH of the formulation is 4.00 to 8.00.

34. T he aqueous pharmaceutical composition accordi ng to C laim 33, wherein pH of the formulation is 6 to 6.60.

35. T he aqueous pharmaceuti cal compositi on accordi ng to the any of the precedi ng claims, wherein the formulation is administered by subcutaneous injection.

36. T he aqueous pharmaceuti cal compositi on accordi ng to the any of the precedi ng claims, wherein the formulation is in a pre filled syringe.