# US 2010/0166774 A1 · Stable Protein Formulations

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- Number: US 2010/0166774 A1
- Document ID: US20100166774A1
- Status: Published application
- Kind: A1
- Application number: 12/086,876
- Filed: 2006-12-19
- Published: 2010-07-01
- Earliest claimed: 2005-12-20
- Assignees: Bristol-Myers Squibb Company
- Inventors: Manisha M. Dali; Charles E. Dahlheim; Sunita Borsadia; Vijay H. Naringrekar; Rajesh Babulal Gandhi; Manoj Nerurkar
- Web page: https://patentgrep.com/patent/US20100166774A1

## CPC

- C07K 14/70521: {CD28, CD152}
- A61K 9/0019: {Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00)}
- A61K 47/26: Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
- A61P 19/02: for joint disorders, e.g. arthritis, arthrosis
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 37/00: Drugs for immunological or allergic disorders
- A61P 37/02: Immunomodulators
- A61P 37/06: Immunosuppressants, e.g. drugs for graft rejection
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- C07K 2319/30: Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

The present invention relates generally to stable formulations comprising CTLA4Ig molecules, including lyophilized, and liquid formulations for administration via various routes including, for example, routes such as intravenous (IV) and subcutaneous (SC) for treating immune system diseases and tolerance induction.

## Claims

### Claim 1

A stable formulation suitable for subcutaneous administration comprising at least 100 mg/ml CTLA4Ig molecules, a sugar capable of stabilizing said formulation at a concentration effective therefore and a pharmaceutically acceptable aqueous carrier.

### Claim 2 (depends on 1)

The formulation of claim 1 wherein the sugar is selected from the group consisting of sucrose, lactose, maltose, mannitol and trehalose and mixtures thereof.

### Claim 3 (depends on 1)

The formulation of claim 1 wherein the sugar is a disaccharide.

### Claim 4 (depends on 3)

The formulation of claim 3 wherein the disaccharide is sucrose.

### Claim 5 (depends on 1)

The formulation of claim 1 further comprising a pharmaceutically acceptable buffer.

### Claim 6 (depends on 1)

The formulation of claim 1 having a pH range of from 6 to 8.

### Claim 7 (depends on 1)

The formulation of claim 1 wherein the CTLA4Ig molecule has the amino acid sequence shown in SEQ ID NO:2 starting at methionine at position 27 or alanine at position 26 and ending at lysine at position 383 or glycine at position 382.

### Claim 8 (depends on 1)

The formulation of claim 1 wherein the CTLA4Ig molecule is L104EA29YIg having the amino acid sequence shown in SEQ ID NO:4 starting at methionine at position 27 or alanine at position 28 and ending at lysine at position 383 or glycine at position 382.

### Claim 9 (depends on 7, 8)

The formulation of claim 7 or 8 wherein the sugar is selected from the group consisting of sucrose, lactose, maltose, mannitol and trehalose and mixtures thereof.

### Claim 10 (depends on 7, 8)

The formulation of claim 7 or 8 wherein the sugar is a disaccharide.

### Claim 11 (depends on 7, 8)

The formulation of claim 7 or 8 wherein the sugar is sucrose.

### Claim 12 (depends on 1, 7, 8)

The solution of claim 1, 7 or 8 wherein the ratio of protein:sugar is at least 1:1.3.

### Claim 13 (depends on 7, 8)

The formulation of claim 7 or 8 further comprising a pharmaceutically acceptable buffer.

### Claim 14 (depends on 7, 8)

The formulation of claim 7 or 8 having a pH range of from 6 to 8.

### Claim 15

A stable formulation comprising the CTLA4Ig molecule having the amino acid sequence shown in SEQ ID NO:2 starting at methionine at position 27 or alanine at position 26 and ending at lysine at position 383 or glycine at position 382 in an amount of about 125 mg/ml, sucrose in an amount of about 170 mg/ml, at least one buffering agent, sterile water for injection and optionally a surfactant.

### Claim 16 (depends on 15)

The formulation of claim 15 wherein the buffering agent is in an amount of at least 10 mM sodium phosphate, wherein the sodium phosphate is a mixture of the dibasic and monobasic salts thereof.

### Claim 17 (depends on 15)

The formulation of claim 15 wherein the surfactant is Poloxamer 188 in an amount of about 8 mg/ml.

### Claim 18 (depends on 15)

The formulation of claim 15 having a pH range of from 6 to 8.

### Claim 19

A stable formulation comprising LE104A29YIg molecule having the amino acid sequence shown in SEQ ID NO:4 starting at methionine at position 27 or alanine at position 26 and ending at lysine at position 383 or glycine at position 382 in an amount of about 125 mg/ml, sucrose in an amount of about 170 mg/ml, at least one buffering agent, sterile water for injection and optionally a surfactant.

### Claim 20 (depends on 19)

The formulation of claim 19 wherein the buffering agent is in an amount of at least 10 mM sodium phosphate, wherein the sodium phosphate is a mixture of the dibasic and monobasic salts thereof.

### Claim 21 (depends on 19)

The formulation of claim 19 wherein the surfactant is Poloxamer 188 in an amount of about 8 mg/ml.

### Claim 22 (depends on 19)

The formulation of claim 19 having a pH range of from 6 to 8.

### Claim 23 (depends on 1, 7, 8, 15, 19)

The formulation of claim 1, 7, 8, 15 or 19 wherein the formulation is stable when stored at 2 to 8 C for at least 12 months.

### Claim 24 (depends on 1, 7, 8, 15, 19)

An article of manufacture comprising:
a) at least one container which holds the formulation of claim 1, 7, 8, 15 or 19 and
b) instructions for administering the formulation subcutaneously to a subject in need thereof.

### Claim 25 (depends on 24)

The article of manufacture of claim 24 wherein the container is a vial or syringe.

### Claim 26

A stable formulation suitable for dilution prior to intravenous administration comprising at least 20 mg/ml of CTLA4Ig molecules, a sugar capable of stabilizing said formulation at a concentration effective therefore and a pharmaceutically acceptable aqueous carrier

### Claim 27 (depends on 26)

The formulation of claim 26 wherein the sugar is selected from the group consisting of sucrose, lactose, maltose, and trehalose, and mixtures thereof

### Claim 28 (depends on 26)

The formulation of claim 26 wherein the sugar is a disaccharide.

### Claim 29 (depends on 26)

The formulation of claim 26 wherein the disaccharide is sucrose.

### Claim 30 (depends on 26)

The formulation of claim 26 further comprising a pharmaceutically acceptable buffer.

### Claim 31 (depends on 26)

The formulation of claim 26 having a pH range of from 6 to 8.

### Claim 32 (depends on 26)

The formulation of claim 26 wherein CTLA4Ig molecule has the amino acid sequence shown in SEQ ID NO:2 starting at methionine at position 27 or alanine at position 26 and ending at lysine at position 383 or glycine at position 382.

### Claim 33 (depends on 26)

The formulation of claim 26 wherein the CTLA4Ig molecule is L104EA29YIg having the amino acid sequence shown in SEQ ID NO:4 starting at methionine at position 27 or alanine at position 26 and ending at lysine at position 383 or glycine at position 382.

### Claim 34 (depends on 32, 33)

The formulation of claim 32 or 33 wherein the sugar is selected from the group consisting of sucrose, lactose, maltose, and trehalose.

### Claim 35 (depends on 32, 33)

The formulation of claim 32 or 33 wherein the sugar is sucrose.

### Claim 36 (depends on 26, 32, 33)

The solution of claim 26, 32 or 33 wherein the ratio of protein:sugar is at least 1:2.

### Claim 37 (depends on 32, 33)

The formulation of claim 32 or 33 further comprising a pharmaceutically acceptable buffer.

### Claim 38 (depends on 32, 33)

The formulation of claim 32 or 33 having a pH range of from 6 to 8.

### Claim 39

A stable formulation comprising CTLA4Ig molecule having the amino acid sequence shown in SEQ ID NO:2 starting at methionine at position 27 or alanine at position 26 and ending at lysine at position 383 or glycine at position 382 in an amount of about 20 mg/ml, sucrose in an amount of about 40 mg/ml, at least one buffering agent, sterile water for injection and optionally a surfactant.

### Claim 40 (depends on 39)

The formulation of claim 39 wherein the buffering agent is in an amount of at least 10 mM sodium phosphate.

### Claim 41 (depends on 39)

The formulation of claim 39 wherein the surfactant is Poloxamer 188 in an amount of about 8 mg/ml.

### Claim 42 (depends on 39)

The formulation of claim 39 having a pH range of from 6 to 8.

### Claim 43

A stable formulation comprising LE104A29YIg molecule having the amino acid sequence shown in SEQ ID NO:4 starting at methionine at position 27 or alanine at position 26 and ending at lysine at position 383 or glycine at position 382 in an amount of about 20 mg/ml, sucrose in an amount of about 40 mg/ml, at least one buffering agent, sterile water for injection and optionally a surfactant.

### Claim 44 (depends on 43)

The formulation of claim 43 wherein the buffering agent is in an amount of at least 10 mM sodium phosphate.

### Claim 45 (depends on 43)

The formulation of claim 43 wherein the surfactant is Poloxamer 188 in an amount of about 8 mg/ml.

### Claim 46 (depends on 43)

The formulation of claim 43 having a pH range of from 6 to 8.

### Claim 47 (depends on 26, 33, 39, 43)

The formulation of claim 26, 33, 33, 39 or 43 wherein the formulation is stable when stored at 2 to 8 C for at least 12 months.

### Claim 48 (depends on 26, 32, 33, 39, 43)

An article of manufacture comprising:
a) at least one container which holds the formulation of claim 26, 32, 33, 39 or 43 and
b) instructions for diluting the formulation with an aqueous carrier to a final CTLA4Ig molecule concentration of from 1 to 10 mg/ml.

### Claim 49 (depends on 48)

The article of manufacture of claim 48 further comprising a syringe.

### Claim 50 (depends on 48)

The article of manufacture of claim 48 where in the aqueous carrier is sterile water for injection, USP or 0.9% sodium chloride injection, USP.

### Claim 51

A stable lyophilized formulation for dilution prior to intravenous administration comprising soluble CTLA4Ig molecule and a lyoprotectant selected from the group consisting of sucrose, lactose, maltose, and trehalose, wherein the lyoprotectant is at a minimum weight ratio of 1:2 protein to lyoprotectant.

### Claim 52 (depends on 51)

The lyophilized formulation of claim 51 wherein the lyoprotectant is sucrose or maltose and mixtures thereof.

### Claim 53 (depends on 51)

The lyophilized formulation of claim 51 further comprising a pharmaceutically acceptable buffer.

### Claim 54 (depends on 51)

The lyophilized formulation of claim 51 having a pH range of from 7 to 8.

### Claim 55 (depends on 51)

The lyophilized formulation of claim 51 wherein CTLA4Ig molecule has the amino acid sequence shown in SEQ ID NO:2 starting at methionine at position 27 or alanine at position 26 and ending at lysine at position 383 or glycine at position 382.

### Claim 56 (depends on 51)

The lyophilized formulation of claim 51 wherein the CTLA4Ig molecule is L104EA29YIg having the amino acid sequence shown in SEQ ID NO:4 starting at methionine at position 27 or alanine at position 26 and ending at lysine at position 383 or glycine at position 382.

### Claim 57 (depends on 55, 56)

The lyophilized formulation of claim 55 or 56 wherein the lyoprotectant is selected from the group consisting of sucrose, lactose, maltose, and trehalose and mixtures thereof.

### Claim 58 (depends on 55, 56)

The lyophilized formulation of claim 55 or 56 wherein the lyoprotectant is sucrose or maltose.

### Claim 59 (depends on 55, 56)

The lyophilized formulation of claim 55 or 56 further comprising a pharmaceutically acceptable buffer.

### Claim 60 (depends on 55, 56)

The formulation of claim 55 or 56 having a pH range of from 7 to 8.

### Claim 61

A stable lyophilized formulation comprising CTLA4Ig molecule having the amino acid sequence shown in SEQ ID NO:2 starting at methionine at position 27 or alanine at position 26 and ending at lysine at position 383 or glycine at position 382 in an amount of about 262 mg/vial, maltose in an amount of about 525 mg/vial, at least one buffering agent, and optionally a surfactant.

### Claim 62 (depends on 61)

The lyophilized formulation of claim 61 wherein the buffering agent is in an amount of at least 10 mM sodium phosphate.

### Claim 63 (depends on 61)

The lyophilized formulation of claim 61 wherein the surfactant is Poloxamer 188 in an amount of about 8 mg/ml.

### Claim 64 (depends on 61)

The lyophilized formulation of claim 61 having a pH range of from 7 to 8.

### Claim 65

A stable lyophilized formulation comprising LE104A29Y molecule having the amino acid sequence shown in SEQ ID NO:4 starting at methionine at position 27 or alanine at position 26 and ending at lysine at position 383 or glycine at position 382 in an amount of about 110 mg/vial, sucrose in an amount of about 220 mg/vial, at least one buffering agent and optionally a surfactant.

### Claim 66 (depends on 65)

The lyophilized formulation of claim 65 wherein the buffering agent is in an amount of at least 10 mM sodium phosphate.

### Claim 67 (depends on 65)

The lyophilized formulation of claim 65 wherein the surfactant is Poloxamer 188 in an amount of about 8 mg/ml.

### Claim 68 (depends on 65)

The lyophilized formulation of claim 65 having a pH range of from 7 to 8.

### Claim 69 (depends on 51, 55, 56, 61, 65)

The lyophilized formulation of claim 51, 55, 56, 61 or 65 wherein the lyophilized formulation is stable when stored at 2 to 8 C for at least 12 months.

### Claim 70 (depends on 51, 55, 56, 61, 65)

An article of manufacture comprising:
a) at least one container which holds the formulation of claim 51, 55, 56, 61 or 65 and
b) instructions for diluting the lyophilized formulation to a final CTLA4Ig molecule concentration of from 1 to 10 mg/ml.

### Claim 71 (depends on 70)

The article of manufacture of claim 70 further comprising a syringe.

### Claim 72 (depends on 70)

The article of manufacture of claim 70 wherein the aqueous carrier is sterile water of injection, USP or 0.9% sodium chloride injection, USP.

### Claim 73 (depends on 1, 7, 8, 15, 19, 26, 32, 33, 39, 43, 51, 55, 56, 61, 65)

A method for treating immune system diseases comprising administering to a subject in need thereof an effective amount of the formulation of any of claims 1, 7, 8, 15, 19, 26, 32, 33, 39, 43, 51, 55, 56, 61, 65.

### Claim 74 (depends on 73)

The method of claim 73 wherein immune system diseases are selected from the group containing autoimmune diseases, immunoproliferative diseases, and graft-related disorders.

### Claim 75 (depends on 1, 7, 8, 15, 19, 26, 32, 33, 39, 43, 51, 55, 56, 61, 65)

A method for treating or preventing graft versus host disease comprising administering to a subject in need thereof an effective amount of the formulation of any of claims 1, 7, 8, 15, 19, 26, 32, 33, 39, 43, 51, 55, 56, 61, 65.

### Claim 76 (depends on 1, 7, 8, 15, 19, 26, 32, 33, 39, 43, 51, 55, 56, 61, 65)

A method for treating rheumatoid arthritis comprising administering to a subject in need thereof an effective amount of the formulation of any of claims 1, 7, 8, 15, 19, 26, 32, 33, 39, 43, 51, 55, 56, 61, 65.

### Claim 77 (depends on 1, 7, 8, 15, 19, 26, 32, 33, 39, 43, 51, 55, 56, 61, 65)

A method for inhibiting solid organ and/or tissue transplant rejections by a subject comprising administering to the subject who received the transplant an effective amount of the formulation of any of claims 1, 7, 8, 15, 19, 26, 32, 33, 39, 43, 51, 55, 56, 61, 65.