Patent Application
20120076779
Biologics are medicinal products created by biological processes, including preparations isolated from natural sources (e.g., human plasma) and recombinant DNA technologies. Within the healthcare and pharmaceutical industries, biologics are becoming increasingly important for patient treatment and overall revenue growth (Goodman M. Nat Rev Drug Discov. (2009) November; 8(11):837). One important class of biologic drugs is therapeutic proteins, both isolated from natural sources and recombinantly produced. For example, plasma proteins are manufactured for therapeutic administration by isolation from pooled human plasma (e.g., GAMMAGARD LIQUID® [IVIG, Immune Globulin Intravenous (Human) 10%]; Baxter International, Deerfield, Ill.) and recombinant means (e.g., ADVATE® [Antihemophilic Factor (Recombinant), Plasma/Albumin-Free Method]; Baxter International, Deerfield, Ill.).
The administration of therapeutic proteins are primarily performed by intravenous (IV), subcutaneous (SQ), and intramuscular administration, although other routes of administration may be used depending upon the therapeutic protein and condition being treated. Most of the immunoglobulins are administered intravenously as larger volumes can be delivered rapidly by the intravenous route to provide the physiologic levels of IgG needed for the effective treatment of various diseases, such as primary immune deficiencies (PID), immune (idiopathic) thrombocytopenic purpura (ITP) and the Kawasaki syndrome. Due to the nature of IV administration, therapy via this route is a slow and timely process, leading to problems with patient compliance.
Subcutaneous (SQ) administration of therapeutic proteins is a convenient alternative to intravenous administration. Compared to IV infusions, SQ administration has several advantages. For example, it can reduce the incidence of systemic reactions, it does not require sometimes-difficult IV access, and gives patients more independence.
In order to improve patient compliance, it would be convenient to provide the protein in a liquid ready to use formulation. However, many human or humanized therapeutic proteins are highly unstable when formulated at or near neutral pH. A variety of degradation pathways exist for proteins especially in liquid formulations, implicating both chemical and physical instability. Chemical instability includes deamination, aggregation, clipping of the peptide backbone, and oxidation of methionine residues. Physical instability encompasses many phenomena, including, for example, aggregation. Protein instability is particularly problematic for labile proteins that are unstable at mildly acidic to neutral pH. To combat these issues, intravenously administrable immunoglobulins have been formulated at acidic pH, effectively increasing their stability in the formulation (products that are formulated at acidic pH are, e.g., Gamunex (Talecris), Gammagard Liquid (Baxter) or Privigen (CSL).
To combat these issues, therapeutic protein compositions are often formulated at acidic pH, effectively increasing their stability in the formulation. Unfortunately, scientific publications have reported that, for example, intramuscular administration of acidic aqueous preparations can cause pain, and potentially could result in tissue damage (Steen et al., 2001; Sluka et al., 2000, the disclosures of which are incorporated by reference herein in their entireties for all purposes). In other cases, where aqueous formulations have been found not to adequately stabilize the therapeutic proteins, lyophilized formulations are used which must be reconstituted prior to administration. In both cases, these factors can cause a less satisfactory drug administration experience and/or inconvenience for the patient, resulting in reduced patient compliance.
As such, there is a need in the art for formulations and methods of formulation that stabilize these labile therapeutic proteins in compositions (e.g., aqueous compositions) at mildly acidic to neutral pH. The present invention satisfies these and other needs by, among other aspects, providing immunoglobulin compositions formulated with histidine at mild acidic to neutral pH that stabilize labile therapeutic proteins.
The present invention is based in part by the surprising finding that the addition of alkali metal salts to aqueous formulations at mildly acidic to neutral pH stabilizes both plasma-derived and recombinant labile therapeutic proteins. As shown by the present studies, immunoglobulins and coagulation factors formulated at mildly acidic to neutral pH with less than 75 mM of an alkali metal chloride salt are highly unstable. However, the addition of greater than 75 mM of an alkali metal chloride salt (e.g., 100 mM or 150 mM sodium chloride) unexpectedly stabilizes these formulations. This finding is contrary to the result of adding alkali metal chloride salts to formulations at acidic pH values, which is shown herein to destabilize these compositions.
Advantageously, the ability to stably formulate labile proteins (e.g., immunoglobulins, coagulation factors, etc.) at mildly acidic and neutral pH allows for the production of therapeutic formulations that are simpler to self-administer. Furthermore, the ability to stably formulate labile therapeutic proteins at mildly acidic to neutral pH allows for the manufacture of pharmaceutical compositions that may be administered subcutaneously (SQ) or intramuscularly (IM) without the pain and potential for tissue damage that is associated with the SQ and IM administration of compositions formulated at acidic pH.
In one aspect, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.5.
In a specific embodiment of the compositions provided above, the composition comprises from 100 mM to 200 mM of an alkali metal chloride salt.
In a specific embodiment of the compositions provided above, the composition comprises from 125 mM to 175 mM of an alkali metal chloride salt.
In a specific embodiment of the compositions provided above, the composition comprises 150±15 mM an alkali metal chloride salt.
In a specific embodiment of the compositions provided above, the alkali metal chloride salt is sodium chloride.
In a specific embodiment of the compositions provided above, the alkali metal chloride salt is potassium chloride.
In a specific embodiment of the compositions provided above, the amino acid is selected from the group consisting of glycine, proline, and histidine.
In a specific embodiment of the compositions provided above, the amino acid is glycine.
In a specific embodiment of the compositions provided above, the amino acid is proline.
In a specific embodiment of the compositions provided above, the amino acid is histidine.
In a specific embodiment of the compositions provided above, the concentration of the amino acid is from 50 mM to 500 mM.
In a specific embodiment of the compositions provided above, the concentration of the amino acid is from 100 mM to 400 mM.
In a specific embodiment of the compositions provided above, the concentration of the amino acid is from 150 mM to 350 mM.
In a specific embodiment of the compositions provided above, the concentration of the amino acid is from 200 mM to 300 mM.
In a specific embodiment of the compositions provided above, the concentration of the amino acid is from 225 mM to 275 mM.
In a specific embodiment of the compositions provided above, the concentration of the amino acid is 250±10 mM.
In a specific embodiment of the compositions provided above, the pH of the composition is from 5.5 to 7.0.
In a specific embodiment of the compositions provided above, the pH of the composition is from 5.5 to 6.5.
In a specific embodiment of the compositions provided above, the pH of the composition is from 5.5 to 6.0.
In a specific embodiment of the compositions provided above, the pH of the composition is from 6.0 to 7.5.
In a specific embodiment of the compositions provided above, the pH of the composition is from 6.0 to 7.0.
In a specific embodiment of the compositions provided above, the pH of the composition is from 6.0 to 6.5.
In a specific embodiment of the compositions provided above, the pH of the composition is from 6.5 to 7.5.
In a specific embodiment of the compositions provided above, the pH of the composition is from 6.5 to 7.0.
In a specific embodiment of the compositions provided above, the pH of the composition is from 7.0 to 7.5.
In a specific embodiment of the compositions provided above, the labile therapeutic protein is a human or humanized protein.
In a specific embodiment of the compositions provided above, the labile therapeutic protein is an immunoglobulin.
In a specific embodiment of the compositions provided above, the immunoglobulin is an IgG immunoglobulin.
In a specific embodiment of the compositions provided above, the immunoglobulin is a polyclonal immunoglobulin.
In a specific embodiment of the compositions provided above, the immunoglobulin is a monoclonal immunoglobulin.
In a specific embodiment of the compositions provided above, the immunoglobulin is a recombinant immunoglobulin.
In a specific embodiment of the compositions provided above, the immunoglobulin is enriched from pooled human plasma.
In a specific embodiment of the compositions provided above, the concentration of the immunoglobulin is 50±5 g/L.
In a specific embodiment of the compositions provided above, the concentration of the immunoglobulin is less than 50 g/L.
In a specific embodiment of the compositions provided above, the concentration of the immunoglobulin is at least 50 g/L.
In a specific embodiment of the compositions provided above, the concentration of the immunoglobulin is from 50 g/L to 150 g/L.
In a specific embodiment of the compositions provided above, the concentration of the immunoglobulin is 100±10 g/L.
In a specific embodiment of the compositions provided above, the concentration of the immunoglobulin is at least 100 g/L.
In a specific embodiment of the compositions provided above, the concentration of the immunoglobulin is 150±15 g/L.
In a specific embodiment of the compositions provided above, the concentration of the immunoglobulin is from 150 g/L to 250 g/L.
In a specific embodiment of the compositions provided above, the concentration of the immunoglobulin is 200±20 g/L.
In a specific embodiment of the compositions provided above, the concentration of the immunoglobulin is at least 200 g/L.
In a specific embodiment of the compositions provided above, at least 95% of the protein in the composition is immunoglobulin.
In a specific embodiment of the compositions provided above, at least 95% of the protein in the composition is IgG immunoglobulin.
In a specific embodiment of the compositions provided above, at least 98% of the protein in the composition is IgG immunoglobulin.
In a specific embodiment of the compositions provided above, the composition is stable for at least 6 months when stored at between about 28° C. and about 32° C.
In a specific embodiment of the compositions provided above, the composition is stable for at least 1 year when stored at between about 28° C. and about 32° C.
In a specific embodiment of the compositions provided above, the composition is stable for at least 2 years when stored at between about 28° C. and about 32° C.
In a specific embodiment of the compositions provided above, the composition is stable for at least 1 month when stored at between about 38° C. and about 42° C.
In a specific embodiment of the compositions provided above, the composition is stable for at least 3 months when stored at between about 38° C. and about 42° C.
In a specific embodiment of the compositions provided above, the composition is stable for at least 6 months when stored at between about 38° C. and about 42° C.
In a specific embodiment of the compositions provided above, the composition is stable for at least 1 year when stored at between about 28° C. and about 32° C.
In a specific embodiment of the compositions provided above, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state is between 0% and 5%.
In a specific embodiment of the compositions provided above, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state is between 0% and 2%.
In a specific embodiment of the compositions provided above, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state is from 0% to 5% and the percentage of immunoglobulin in the monomeric state is from 80% to 100%.
In a specific embodiment of the compositions provided above, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state is from 0% to 2% and the percentage of immunoglobulin in the monomeric state is from 85% to 100%.
In a specific embodiment of the compositions provided above, the labile therapeutic protein is a coagulation factor.
In a specific embodiment of the compositions provided above, the coagulation factor is selected from the group consisting of Factor VII, Factor VIII, Factor IX, and von Willebrand Factor (vWF).
In a specific embodiment of the compositions provided above, the coagulation factor is Factor VIII.
In a specific embodiment of the compositions provided above, the pH of the composition is between about 6.0 and about 7.0.
In a specific embodiment of the compositions provided above, the pH of the composition is 6.5±0.2.
In a specific embodiment of the compositions provided above, the composition retains at least 80% of its Factor VIII activity when stored at a temperature between about 2° C. and about 8° C. for at least 1 month.
In a specific embodiment of the compositions provided above, the composition retains at least 80% of its Factor VIII activity when stored at a temperature between about 2° C. and about 8° C. for at least 2 months.
In a specific embodiment of the compositions provided above, the composition retains at least 80% of its Factor VIII activity when stored at a temperature between about 2° C. and about 8° C. for at least 3 months.
In a specific embodiment of the compositions provided above, the coagulation factor is Factor VII.
In a specific embodiment of the compositions provided above, the coagulation factor is Factor IX.
In a specific embodiment of the compositions provided above, the coagulation factor is von Willebrand Factor (vWF).
In a specific embodiment of the compositions provided above, the coagulation factor is a protein K-dependent coagulation complex.
In a specific embodiment of the compositions provided above, the protein K-dependent coagulation complex comprises the coagulation factors Factor II, Factor IX, and Factor X.
In a specific embodiment of the compositions provided above, the protein K-dependent coagulation complex further comprises Factor VII.
In a specific embodiment of the compositions provided above, the labile protein is stable for less than 3 months in an aqueous formulation containing: from 0 mM to 50 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.
In a specific embodiment of the compositions provided above, the labile protein is stable for less than 2 months in an aqueous formulation containing: from 0 mM to 50 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.
In a specific embodiment of the compositions provided above, the labile protein is stable for less than 1 month in an aqueous formulation containing: from 0 mM to 50 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.
In a specific embodiment of the compositions provided above, the labile protein is stable for less than 2 weeks in an aqueous formulation containing: (a) from 0 mM to 50 mM of an alkali metal chloride salt; (b) an amino acid; and (c) a pH of from 5.5 to 7.
In a specific embodiment of the compositions provided above, the composition is formulated for subcutaneous and/or intramuscular administration.
In another aspect, the present invention provides a method for stabilizing an aqueous composition of a labile therapeutic protein, the method comprising formulating the composition at a pH between 5.5 and 7.0, wherein the formulated composition comprises: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; and an amino acid.
In a specific embodiment of the methods provided above, the formulated composition comprises from 100 mM to 200 mM of an alkali metal chloride salt.
In a specific embodiment of the methods provided above, the formulated composition comprises from 125 mM to 175 mM of an alkali metal chloride salt.
In a specific embodiment of the methods provided above, the formulated composition comprises 150±15 mM an alkali metal chloride salt.
In a specific embodiment of the methods provided above, the alkali metal chloride salt is sodium chloride.
In a specific embodiment of the methods provided above, the alkali metal chloride salt is potassium chloride.
In a specific embodiment of the methods provided above, the amino acid is selected from the group consisting of glycine, proline, and histidine.
In a specific embodiment of the methods provided above, the amino acid is glycine.
In a specific embodiment of the methods provided above, the amino acid is proline.
In a specific embodiment of the methods provided above, the amino acid is histidine.
In a specific embodiment of the methods provided above, the formulated composition comprises from 50 mM to 500 mM of the amino acid.
In a specific embodiment of the methods provided above, the formulated composition comprises from 100 mM to 400 mM of the amino acid.
In a specific embodiment of the methods provided above, the formulated composition comprises from 150 mM to 350 mM of the amino acid.
In a specific embodiment of the methods provided above, the formulated composition comprises from 200 mM to 300 mM of the amino acid.
In a specific embodiment of the methods provided above, the formulated composition comprises from 225 mM to 275 mM of the amino acid.
In a specific embodiment of the methods provided above, the formulated composition comprises from 250±10 mM of the amino acid.
In a specific embodiment of the methods provided above, the pH of the formulated composition is from 5.5 to 7.0.
In a specific embodiment of the methods provided above, the pH of the formulated composition is from 5.5 to 6.5.
In a specific embodiment of the methods provided above, the pH of the formulated composition is from 5.5 to 6.0.
In a specific embodiment of the methods provided above, the pH of the formulated composition is from 6.0 to 7.5.
In a specific embodiment of the methods provided above, the pH of the formulated composition is from 6.0 to 7.0.
In a specific embodiment of the methods provided above, the pH of the formulated composition is from 6.0 to 6.5.
In a specific embodiment of the methods provided above, the pH of the formulated composition is from 6.5 to 7.5.
In a specific embodiment of the methods provided above, the pH of the formulated composition is from 6.5 to 7.0.
In a specific embodiment of the methods provided above, the pH of the formulated composition is from 7.0 to 7.5.
In a specific embodiment of the methods provided above, the labile therapeutic protein is a human or humanized protein.
In a specific embodiment of the methods provided above, the labile therapeutic protein is an immunoglobulin.
In a specific embodiment of the methods provided above, the immunoglobulin is an IgG immunoglobulin.
In a specific embodiment of the methods provided above, the immunoglobulin is a polyclonal immunoglobulin.
In a specific embodiment of the methods provided above, the immunoglobulin is a monoclonal immunoglobulin.
In a specific embodiment of the methods provided above, the immunoglobulin is a recombinant immunoglobulin.
In a specific embodiment of the methods provided above, the immunoglobulin is enriched from pooled human plasma.
In a specific embodiment of the methods provided above, the concentration of the immunoglobulin is 50±5 g/L.
In a specific embodiment of the methods provided above, the concentration of the immunoglobulin is less than 50 g/L.
In a specific embodiment of the methods provided above, the concentration of the immunoglobulin is at least 50 g/L.
In a specific embodiment of the methods provided above, the concentration of the immunoglobulin is from 50 g/L to 150 g/L.
In a specific embodiment of the methods provided above, the concentration of the immunoglobulin is 100±10 g/L.
In a specific embodiment of the methods provided above, the concentration of the immunoglobulin is at least 100 g/L.
In a specific embodiment of the methods provided above, the concentration of the immunoglobulin is 150±15 g/L.
In a specific embodiment of the methods provided above, the concentration of the immunoglobulin is from 150 g/L to 250 g/L.
In a specific embodiment of the methods provided above, the concentration of the immunoglobulin is 200±20 g/L.
In a specific embodiment of the methods provided above, the concentration of the immunoglobulin is at least 200 g/L.
In a specific embodiment of the methods provided above, at least 95% of the protein in the formulated composition is immunoglobulin.
In a specific embodiment of the methods provided above, at least 95% of the protein in the formulated composition is IgG immunoglobulin.
In a specific embodiment of the methods provided above, at least 98% of the protein in the formulated composition is IgG immunoglobulin.
In a specific embodiment of the methods provided above, the formulated composition is stable for at least 6 months when stored at between about 28° C. and about 32° C.
In a specific embodiment of the methods provided above, the formulated composition is stable for at least 1 year when stored at between about 28° C. and about 32° C.
In a specific embodiment of the methods provided above, the formulated composition is stable for at least 2 years when stored at between about 28° C. and about 32° C.
In a specific embodiment of the methods provided above, the formulated composition is stable for at least 1 month when stored at between about 38° C. and about 42° C.
In a specific embodiment of the methods provided above, the formulate composition is stable for at least 3 months when stored at between about 38° C. and about 42° C.
In a specific embodiment of the methods provided above, the formulated composition is stable for at least 6 months when stored at between about 38° C. and about 42° C.
In a specific embodiment of the methods provided above, the formulated composition is stable for at least 1 year when stored at between about 28° C. and about 32° C.
In a specific embodiment of the methods provided above, the formulated composition is considered stable as long as the percentage of immunoglobulin in the aggregated state is between 0% and 5%.
In a specific embodiment of the methods provided above, the formulated composition is considered stable as long as the percentage of immunoglobulin in the aggregated state is between 0% and 2%.
In a specific embodiment of the methods provided above, the formulated composition is considered stable as long as the percentage of immunoglobulin in the aggregated state is from 0% to 5% and the percentage of immunoglobulin in the monomeric state is from 80% to 100%.
In a specific embodiment of the methods provided above, the formulated composition is considered stable as long as the percentage of immunoglobulin in the aggregated state is from 0% to 2% and the percentage of immunoglobulin in the monomeric state is from 85% to 100%.
In a specific embodiment of the methods provided above, the labile therapeutic protein is a coagulation factor.
In a specific embodiment of the methods provided above, the coagulation factor is selected from the group consisting of Factor VII, Factor VIII, Factor IX, and von Willebrand Factor (vWF).
In a specific embodiment of the methods provided above, the coagulation factor is Factor VIII.
In a specific embodiment of the methods provided above, the pH of the composition is between about 6.0 and about 7.0.
In a specific embodiment of the methods provided above, the pH of the composition is 6.5±0.2.
In a specific embodiment of the methods provided above, the formulated composition retains at least 80% of its Factor VIII activity when stored at a temperature between about 2° C. and about 8° C. for at least 1 month.
In a specific embodiment of the methods provided above, the formulated composition retains at least 80% of its Factor VIII activity when stored at a temperature between about 2° C. and about 8° C. for at least 2 months.
In a specific embodiment of the methods provided above, the formulated composition retains at least 80% of its Factor VIII activity when stored at a temperature between about 2° C. and about 8° C. for at least 3 months.
In a specific embodiment of the methods provided above, the coagulation factor is Factor VII.
In a specific embodiment of the methods provided above, the coagulation factor is Factor IX.
In a specific embodiment of the methods provided above, the coagulation factor is von Willebrand Factor (vWF).
In a specific embodiment of the methods provided above, the coagulation factor is a protein K-dependent coagulation complex.
In a specific embodiment of the methods provided above, the protein K-dependent coagulation complex comprises the coagulation factors Factor II, Factor IX, and Factor X.
In a specific embodiment of the methods provided above, the protein K-dependent coagulation complex further comprises Factor VII.
In a specific embodiment of the methods provided above, the labile protein is stable for less than 3 months in an aqueous formulation containing: from 0 mM to 50 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.
In a specific embodiment of the methods provided above, the labile protein is stable for less than 2 months in an aqueous formulation containing: from 0 mM to 50 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.
In a specific embodiment of the methods provided above, the labile protein is stable for less than 1 month in an aqueous formulation containing: from 0 mM to 50 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.
In a specific embodiment of the methods provided above, the labile protein is stable for less than 2 weeks in an aqueous formulation containing: from 0 mM to 50 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.
In a specific embodiment of the methods provided above, the composition is formulated for subcutaneous and/or intramuscular administration.
Therapeutic proteins are often times formulated at acidic pH or as lyophilized compositions due to their labile nature in aqueous solution at or near neutral pH. As discussed above, these formulations are less convenient, may cause pain and/or tissue damage upon administration, and likely reduce patient compliance. Advantageously, the present invention provides means for stably formulating these labile proteins in aqueous solution at or near neutral pH. In one aspect, the present invention provides labile therapeutic protein compositions stabilized by the addition of moderate levels of alkali metal chloride salts (e.g., 75 mM to 200 mM, preferably 100 mM to 200 mM) to formulations at mildly acidic to neutral pH. The present invention is based in part on the surprising discovery that labile therapeutic proteins are significantly stabilized at mildly acidic to neutral pH by the addition of an alkali metal chloride salt at a final concentration of between about 75 mM and about 200 mM.
Our new studies provided herein demonstrate that purified plasma-derived immunoglobulin preparations formulated in 0.25 M glycine could be stabilized by the addition of sodium chloride in a pH dependent manner. Examples 1 and 2 shows that these immunoglobulin preparations, having a final concentration of between about 90 g/L and about 220 g/L, were stabilized for at least 24 months when stored at a temperature of 28° C. to 32° C., and for at least 6 months when stored at a temperature of 38° C. to 42° C. Maximum stability was observed with addition of 150 mM sodium chloride.
Under these conditions, the addition of sodium chloride to formulations at pH values at and above 7.0 resulted in considerably higher aggregation and fragmentation rates, compared to samples formulated at a pH between 5.5 and 7.0 (Table 5 and Table 6). Similarly, it was previously observed that sodium chloride significantly destabilized immunoglobulin formulations with acidic pH values (under 5.0).
Similar stabilizing effects were demonstrated with the addition of 150 mM sodium chloride to two hyper-immune immunoglobulin preparations, Partobulin® NG and Tetabulin® NG. Example 3 demonstrates that addition of 150 mM sodium chloride to these plasma-derived immunoglobulin preparations results in reduced protein aggregation at pH values between 5.5 and 6.5 and reduced loss of anti-D titer for Partobulin® NG between pH 5.5 and pH 6.5, while stabilizing the Tetanus anti-toxin titer across the entire range of pH investigated.
Next, it was demonstrated that sodium chloride would also stabilize recombinant antibody preparations formulated at mildly acidic to neutral pH. Example 4 shows that a recombinant anti-MIF antibody formulated at pH 5.6 to 6.5 with 0.25 M glycine and 150 mM sodium chloride Was stabilized upon storage at elevated temperatures (38° C. to 42° C.) for six months. Consistent with the previous observations, sodium chloride enhanced degradation when the antibody was formulated at low pH (4.5) and increased aggregation when formulated at a higher pH (pH 7.3).
Finally, to investigate whether or not the stabilizing effect of sodium chloride at mildly acidic to neutral pH can be applied to non-immunoglobulin labile therapeutic proteins, formulations of recombinant Factor VIII (rFVIII) were prepared. As the foundation for the rFVIII formulation, the protein was formulated as in the reconstituted ADVATE® (Baxter International; Deerfield Ill.) product. Despite the presence of several traditional stabilizing agents, including mannitol, trehalose, histidine, calcium chloride, polysorbate-80 and glutathione, rFVIII is extremely unstable, even at 2° C. to 8° C., in aqueous formulation. As such, ADVATE® is marketed as a lyophilized formulation that is reconstituted immediately prior to administration.
Remarkably, as shown in Example 5, the inclusion of 150 mM sodium chloride almost completely stabilizes rFVIII activity at pH 6.0 to 7.0, when stored in aqueous formulation at 2° C. to 8° C. for at least 12 weeks. Accordingly, it has now been demonstrated that intermediate levels of an alkali metal chloride salt can stabilize a wide range of labile therapeutic proteins when formulated at a mildly acidic to neutral pH.
As used herein, a “labile therapeutic protein” refers to a class of therapeutically useful proteins that are unstable when formulated at mildly acid to neutral pH in the absence of an alkali metal chloride salt. Generally, protein stability can be measured by several different metrics, including aggregation, loss of enzymatic activity, loss of antigenic titer, or degradation. Labile therapeutic proteins will display one or more of these unwanted characteristics when stored at mildly acidic to neutral pH in the absence of an alkali metal chloride salt. The absolute time for which a labile protein is stable will be dependent upon the individual characteristics of the protein, which can be readily determined by the skilled artisan. For example, certain blood coagulations proteins (e.g., Factor VIII) are stable for less than two months under refrigeration when formulated a mildly acidic to neutral pH in the absence of suitable levels of an alkali metal chloride salt. In other cases, for example plasma derived immunoglobulin preparations, a labile therapeutic protein may be stable at room temperature for less than six months when formulated a mildly acidic to neutral pH in the absence of suitable levels of an alkali metal chloride salt, as compared to more than two years in the presence of moderate levels of an alkali metal chloride salt.
In the context of the present invention, a labile protein will show a marked increase in stability upon the addition of an alkali metal chloride salt to a formulation at mildly acid to neutral pH. Inclusion of an alkali metal chloride salt may, for example, reduce aggregation of the labile protein by at least about 20%; maintain at least about 20% more enzymatic activity; maintain at least about 20% more antigenic titer; and/or reduce degradation by at least about 20%, when stored for a given period of time. For the purposes of the present invention, a labile therapeutic protein may be isolated from a natural source (e.g., plasma-derived) or recombinantly produced. For example, in certain embodiments, labile plasma derived blood proteins, such as immunoglobulins, and blood coagulation factors (e.g., Factor VIII) are particularly well suited for formulation as described herein. Non-limiting examples of coagulation proteins include, Factor II (prothrombin), Factor III (platelet tissue factor), Factor V, Factor VII, Factor VIII, Factor IX, Factor X, Factor XI, Factor XII, Factor XIII, von Willebrand Factor (vWF). Likewise, labile recombinant proteins, such as antibodies and blood coagulation factors may be stabilized according to the formulations and methods provided herein.
As used herein, a “storage stable” aqueous composition refers to a protein solution that has been formulated to increase the stability of the protein in solution, for example by at least 20%, over a given storage time. In the context of the present invention, a labile protein solution formulated at a mildly acidic to neutral pH can be made “storage stable” by the addition of a moderate level (about 75 mM to about 200 mM, preferably about 100 mM to about 200 mM) of an alkali metal chloride salt. The stability of the protein in any given formulation can be measured, for example, by monitoring the formation of aggregates, loss of bulk enzymatic activity, loss of antigenic titer or formation of degradation products, over a period of time. The absolute stability of a formulation, and the stabilizing effects of the alkali metal chloride salt, will vary dependent upon the labile protein being stabilized.
As used herein, the term “time of stability” refers to the length of time a composition is considered stable. For example, the time of stability for a composition may refer to the length of time for which the level of protein aggregation and/or degradation in the composition remains below a certain threshold (e.g., 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc.), the length of time a composition maintains an enzymatic activity above a certain threshold (e.g., 100%, 95%, 90, 85, 80, 75, 70, 65, 60, 55, 50, etc. of the amount of activity present in the composition at the start of the storage period), or the length of time a composition maintains an antigenic titer (e.g., 100%, 95%, 90, 85, 80, 75, 70, 65, 60, 55, 50, etc. of the antigenic titer present in the composition at the start of the storage period). In the context of the present invention, a storage stable aqueous composition of a labile therapeutic protein formulated at mildly acidic to neutral pH with a moderate level of an alkali metal chloride salt will have a longer time of stability than a composition of the same labile therapeutic protein formulated at mildly acidic to neutral pH without a moderate level of an alkali metal chloride salt. A storage stable aqueous composition of a labile therapeutic protein, as provided herein, will have a time of stability that is, for example, at least 20% greater than the time of stability for the same composition formulated in the absence of a moderate level of an alkali metal chloride salt, or at least 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190% greater, or at least 2 times greater, or at least 2.5, 3.0 times, 3.5 times, 4.0 times, 4.5 times, 5.0 times, 5.5 times, 6.0 times, 6.5 times, 7.0 times, 7.5 times, 8.0 times, 8.5 times, 9.0 times, 9.5 times, 10 times, or more times greater than the time of stability for the same composition formulated in the absence of a moderate level of an alkali metal chloride salt.
As used herein, the term “stable” refers to a state of a protein composition (e.g., an immunoglobulin solution) suitable for pharmaceutical administration. In the context of the present invention, an immunoglobulin solution is generally considered to be stable when the level of immunoglobulin aggregation and/or degradation in the composition remains below a certain threshold (e.g., below 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc.) or the when the antigenic titer remains above a certain threshold (e.g., above 100%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, etc. of the antigenic titer present in the composition at the start of a storage period).
The European Pharmacopoeia (Ph. Eur.) standard for normal human immunoglobulins is that the composition has: (i) a monomer and dimer content equal to or greater than 85% of the total area of a standard chromatogram; and (ii) a polymer and aggregate sum content of not more than 10% of the total area of the chromatogram. For IGIV the sum of the peak areas of the monomer and dimer represents not less than 95 percent and the maximum amount of polymers and aggregates is no more than 2%. Accordingly, in one embodiment, a storage-stable immunoglobulin composition provided herein is considered to be stable when at least 85% of the immunoglobulin content is monomeric and no more than 5%, preferably no more than 2%, of the immunoglobulin content is aggregated.
As used herein, “storage” means that a formulation is not immediately administered to a subject once prepared, but is kept for a period of time under particular conditions (e.g. at a particular temperature, under a particular atmosphere, protected from light, etc.) prior to use. For example, a liquid formulation can be kept for days, weeks, months or years, prior to administration to a subject under varied temperatures such as refrigerated (0° to 10° C.) or room temperature (e.g., between about 20° C. and 25° C.).
For the purposes of the present invention, when referring to a concentration of an individual component of a composition, the phrases “no more than X” and “from 0 to X” are equivalent and refer to any concentration between and including 0 and X. For example, the phrases “a concentration of no more than 2%” and “a concentration of from 0% to 2%” are equivalent and include 0%, 1%, and 2%.
For the purposes of the present invention, when referring to a concentration of an individual component of a composition, the phrases “no less than X” refers to any concentration X or higher. For example, the phrase “a concentration of no less than 98%” includes 98%, 99%, and 100%.
For the purposes of the present invention, when referring to a concentration of an individual component of a composition, the phrases “between X and Y” and “from X to X” are equivalent and refer to any concentration between and including X and Y. For example, the phrases “a concentration of between 49% and 51%” and “a concentration of from 49% to 51%” are equivalent and include 49%, 50%, and 51%.
As used herein, an “alkali metal chloride salt” refers to an inorganic salt of an alkali metal and chlorine. For the purposes of the present invention, the alkali metal chloride salt will be a pharmaceutically acceptable salt, most commonly sodium or potassium chloride. In a preferred embodiment, the salt is sodium chloride.
Likewise, an “alkali metal cation” will most commonly refer to a sodium cation (Na+) or potassium cation (K+) and can be contributed by an alkali metal chloride salt or other source. In the context of the present invention, a hydrogen ion is not considered an alkali metal cation, and thus the inclusion of hydrochloric acid will not contribute to the alkali metal cation content of the formulation.
As used herein, a “coagulation factor” refers to a protein involved in the intrinsic (contact activation) or extrinsic (tissue factor) pathway of the coagulation cascade. Non-limiting examples of coagulation proteins include, Factor II (prothrombin), Factor III (platelet tissue factor), Factor. V, Factor VII, Factor VIII, Factor IX, Factor X, Factor XI, Factor XII, Factor XIII, von Willebrand Factor (vWF), and the like. Coagulation proteins stabilized by the formulations and methods provided herein may be plasma-derived or recombinantly produced.
As used herein, the term “core coagulation factor” refers to any one of Factor VII, Factor VIII, Factor IX, and von Willebrand Factor (vWF), as well as conservative or natural variants, biologically active fragments, and natural isoforms thereof.
As used herein, the term “Factor VIII” or “FVIII” refers to any form of factor VIII molecule with the typical characteristics of blood coagulation factor VIII, whether derived from blood plasma or produced through the use of recombinant DNA techniques, and including all modified forms of factor VIII. Factor VIII (FVIII) exists naturally and in therapeutic preparations as a heterogeneous distribution of polypeptides arising from a single gene product (see, e.g., Anderson et al., Proc. Natl. Acad. Sci. USA, 83:2979-2983 (1986)). Commercially available examples of therapeutic preparations containing Factor VIII include those sold under the trade names of HEMOFIL M, ADVATE, and RECOMBINATE (available from Baxter Healthcare Corporation, Deerfield, Ill., U.S.A.).
As used herein, an “antibody” refers to a polypeptide substantially encoded by an immunoglobulin gene or immunoglobulin genes, or fragments thereof, which specifically bind and recognize an analyte (antigen). The recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon and mu constant region genes, as well as the myriad immunoglobulin variable region genes. Light chains are classified as either kappa or lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD, and IgE, respectively.
An exemplary immunoglobulin (antibody) structural unit is composed of two pairs of polypeptide chains, each pair having one “light” (about 25 kD) and one “heavy” chain (about 50-70 kD). The N-terminus of each chain defines a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The terms variable light chain (VL) and variable heavy chain (VH) refer to these light and heavy chains respectively. In a particular exemplary embodiment, the immunoglobulin will consist of an immunoglobulin preparation isolated from pooled plasma (preferably human plasma) comprising IgG immunoglobulins.
As used herein, the term “about” denotes an approximate range of plus or minus 10% from a specified value. For instance, the language “about 20%” encompasses a range of 18-22%. As used herein, about also includes the exact amount. Hence “about 20%” means “about 20%” and also “20%.”
By “therapeutically effective amount or dose” or “sufficient/effective amount or dose,” it is meant a dose that produces effects for which it is administered. The exact dose will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins).
As used herein, a “stabilizing agent” refers to a chemical, other than an alkali metal chloride salt, which aids in the stabilization of a labile therapeutic agent in an aqueous formulation under mildly acid to neutral pH. Examples of suitable stabilizing agents for the formulations and methods provided herein include, without limitation, buffering agents (e.g., TRIS, HEPES, potassium or sodium phosphate, amino acids, etc.), osmolytes (e.g., sugars, sugar alcohols, etc.), bulking agents (e.g., amino acids, etc.), divalent salts, surfactants, and the like.
As used herein, “amino acids” refers to any natural or non-natural pharmaceutically acceptable amino acid. Non-limiting examples of amino acids include, isoleucine, alanine, leucine, asparagine, lysine, aspartic acid, methionine, cysteine, phenylalanine, glutamic acid, threonine, glutamine, tryptophan, glycine, valine, proline, selenocysteine, serine, tyrosine, arginine, histidine, ornithine, taurine, and the like.
Any sugar such as mono-, di-, or polysaccharides, or water-soluble glucans, including for example fructose, glucose, mannose, sorbose, xylose, maltose, lactose, sucrose, dextran, trehalose, pullulan, dextrin, cyclodextrin, soluble starch, hydroxyethyl starch, and carboxymethylcellulose may be used.
As used herein, a “sugar alcohol” refers to a hydrocarbon having between about 4 and about 8 carbon atoms and at least one hydroxyl group. Non-limiting examples of sugar alcohols that may be used in formulations provided herein include, mannitol, sorbitol, inositol, galactitol, dulcitol, xylitol, and arabitol.
As used herein, the term “activity” refers to a functional activity or activities of a polypeptide or portion thereof associated with a full-length (complete) protein. Functional activities include, but are not limited to, biological activity, catalytic or enzymatic activity, antigenicity (i.e., the ability to bind or compete with a polypeptide for binding to an anti-polypeptide antibody), immunogenicity, ability to form multimers, and the ability to specifically bind to a receptor or ligand for the polypeptide.
Among other aspects, the present invention provides stabilized formulations of labile proteins for therapeutic administration. The following embodiments are based in part on the unexpected discovery that the addition of moderate levels of an alkali metal chloride salt (i.e., about 75 mM to about 200 mM, preferably about 100 mM to about 200 mM) to formulations at mildly acidic to neutral pH stabilize various plasma-derived and recombinant proteins that are otherwise labile at these pH values.
The labile therapeutic protein compositions provided by the present invention take advantage of the increased stability afforded when these proteins are formulated at mildly acidic to neutral pH. Generally, this includes pH values between about 5.5 and about 7.5. In a preferred embodiment, the pH value is between about 5.5 and about 7.0. However, the range of pH values at which any individual labile therapeutic protein is stabilized by the addition of a moderate level (i.e., between about 75 mM and about 200 mM, preferably between about 100 mM and about 200 mM) of an alkali metal chloride salt may vary slightly, dependent upon the properties of the individual protein. For example, in one embodiment, a storage stable formulation will have a pH between about 5.5 and about 7.0. In another embodiment, a storage stable formulation will have a pH between about 5.5 and about 6.5. In other embodiments, the pH of the stabilizing formulation will be between about 6.0 and about 7.0. In another embodiment, the pH of the stabilizing formulation will be between about 5.5 and about 6.0. In one embodiment, the pH of the stabilizing formulation will be between about 6.0 and about 6.5. In another embodiment, the pH of the stabilizing formulation will be between about 6.5 and about 7.0. In another embodiment, a storage stable formulation will have a pH between about 6.0 and about 7.5. In another embodiment, a storage stable formulation will have a pH between about 6.5 and about 7.5. In another embodiment, a storage stable formulation will have a pH between about 7.0 and about 7.5. In other embodiments, the pH of the stabilizing formulation is 5.5±0.2, 5.6±0.2, 5.7±0.2, 5.8±0.2, 5.9±0.2, 6.0±0.2, 6.1±0.2, 6.2±0.2, 6.3±0.2, 6.4±0.2, 6.5±0.2, 6.6±0.2, 6.7±0.2, 6.8±0.2, 6.9±0.2, 7.0±0.2, 7.1±0.2, 7.2±0.2, 7.3±0.2, 7.4±0.2, or 7.5±0.2. In other embodiments, the pH of the stabilizing formulation is 5.5±0.1, 5.6±0.1, 5.7±0.1, 5.8±0.1, 5.9±0.1, 6.0±0.1, 6.1±0.1, 6.2±0.1, 6.3±0.1, 6.4±0.1, 6.5±0.1, 6.6±0.1, 6.7±0.1, 6.8±0.1, 6.9±0.1, 7.0±0.1, 7.1±0.1, 7.2±0.1, 7.3±0.1, 7.4±0.1, or 7.5±0.1. In yet other embodiments, the pH of the stabilizing formulation is 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, or 7.5.
In addition to the alkali metal chloride salt, the pharmaceutical compositions provided herein include one or more stabilizing agents. In a specific embodiment, the stabilizing agent is a buffering agent suitable for intravenous, intravitreal, subcutaneous, and/or intramuscular administration. Non-limiting examples of buffering agents suitable for formulating the storage stable compositions provided herein include glycine, histidine, proline, or other amino acids, salts like citrate, phosphate, acetate, glutamate, tartrate, benzoate, lactate, gluconate, malate, succinate, formate, propionate, carbonate, or any combination thereof adjusted to an appropriate pH. Generally, the buffering agent will be sufficient to maintain a suitable pH in the formulation for an extended period of time.
Among other aspects, the present invention provides stabilized formulations of labile therapeutic proteins for therapeutic administration. The following embodiments are based in part on the unexpected discovery that the formulation of labile therapeutic proteins, including but not limited to immunoglobulin and coagulation factors, with a moderate amount of an alkali metal chloride salt at mildly acidic to neutral pH stabilizes proteins that are otherwise labile at these pH values and/or labile when formulated with an alkali metal chloride salt an acidic pH.
As will be recognized by one of skill in the art, the formulation of a protein composition at a particular pH may introduce residual counter ions contributed from one or more pH modifying agents. For example, the storage stable compositions provided herein may contain chloride anions contributed from hydrochloric acid, acetate anions contributed from glacial acetic acid, sodium cations contributed from sodium hydroxide, and the like. In the context of the present invention, a storage stable labile therapeutic protein composition consisting of or consisting essentially of: a labile therapeutic protein, a moderate concentration of an alkali metal chloride salt, and a stabilizing agent may further comprise one or more counter ion, as necessitated by the formulation process at the desired pH.
In one embodiment, a labile therapeutic protein that may benefit from a formulation or method provided herein is stable for less than 3 months at a temperature between 28° C. and 32° C. when in an aqueous formulation containing less than 50 mM of an alkali metal chloride salt at a pH between 5.5 and 7.5, preferably between 5.5 and 7.0. In another embodiment, the labile therapeutic protein is stable for less than 2 months at a temperature between 28° C. and 32° C. in an aqueous formulation containing less than 50 mM of an alkali metal chloride salt at a pH between 5.5 and 7.5, preferably between 5.5 and 7.0. In another embodiment, the labile therapeutic protein is stable for less than 1 month at a temperature between 28° C. and 32° C. in an aqueous formulation containing less than 50 mM of an alkali metal chloride salt at a pH between 5.5 and 7.5, preferably between 5.5 and 7.0. In yet another embodiment, the labile therapeutic protein is stable for less than 2 weeks at a temperature between 28° C. and 32° C. in an aqueous formulation containing less than 50 mM of an alkali metal chloride salt at a pH between 5.5 and 7.5, preferably between 5.5 and 7.0. In a preferred embodiment, the labile therapeutic protein is an immunoglobulin.
In a related embodiment, a labile therapeutic protein that may benefit from a formulation or method provided herein is stable for less than 3 months at a temperature between 2° C. and 8° C. when in an aqueous formulation containing less than 50 mM of an alkali metal chloride salt at a pH between 5.5 and 7.5, preferably between 6.0 and 7.5. In another embodiment, the labile therapeutic protein is stable for less than 2 months at a temperature between 2° C. and 8° C. in an aqueous formulation containing less than 50 mM of an alkali metal chloride salt at a pH between 5.5 and 7.5, preferably between 6.0 and 7.5. In another embodiment, the labile therapeutic protein is stable for less than 1 month at a temperature between 2° C. and 8° C. in an aqueous formulation containing less than 50 mM of an alkali metal chloride salt at a pH between 5.5 and 7.5, preferably between 6.0 and 7.5. In yet another embodiment, the labile therapeutic protein is stable for less than 2 weeks at a temperature between 2° C. and 8° C. in an aqueous formulation containing less than 50 mM of an alkali metal chloride salt at a pH between 5.5 and 7.5, preferably between 6.0 and 7.5. In a specific embodiment, the labile therapeutic protein is a coagulation factor. In a preferred embodiment, the labile therapeutic protein is Factor VIII.
In preferred embodiments of the invention, the labile therapeutic protein will be a human protein, a chimeric protein (e.g., a mouse/human chimera or rat/human chimera), or a humanized protein. For example, in one preferred embodiment, the labile therapeutic protein will be a recombinant chimeric or humanized monoclonal antibody. In other preferred embodiment, the labile therapeutic protein will be a plasma protein, either recombinant or plasma-derived, e.g., a protein composition isolated from pooled human plasma. Non-limiting examples of plasma-derived labile proteins that may be formulated according to the present invention include: Factor II (prothrombin), Factor III (platelet tissue factor), Factor V, Factor VII, Factor VIII, Factor IX, Factor X, Factor XI, Factor XII, Factor XIII, and von Willebrand Factor (vWF).
The final concentration of the labile therapeutic protein in the formulations provided herein will be dependent upon many factors, including without limitation, the potency and specific activity of the protein, the disease or condition being treated, the route of administration, and other factors that will be well understood by the skilled practitioner. In one embodiment, the labile therapeutic protein will be formulated at a low protein concentration, for example, between 0.05 mg/mL and 10 mg/mL, or between 0.1 mg/mL and 20 mg/mL, or between 0.5 mg/mL and 10 mg/mL, or between 0.1 mg/mL and 0.5 mg/mL. In other embodiments, the labile therapeutic protein will be formulated at a moderate protein concentration, between 20 mg/mL and 80 mg/mL. For example, between 20 mg/mL and 40 mg/mL. Or between 40 mg/mL and 60 mg/mL. Or between 60 mg/mL and 80 mg/mL. In yet other embodiments, the labile therapeutic protein will be formulated at a high protein concentration, between 80 mg/mL and 250 mg/mL. For example, in one embodiment, the protein concentration will be between 80 mg/mL and 120 mg/mL. In another embodiment, the protein concentration will be between 120 mg/mL and 180 mg/mL. In yet another embodiment, the protein concentration will be between 180 mg/mL and 250 mg/mL.
In certain embodiments, the final protein concentration may be between 0.5% and 25%. In another embodiment, the final protein concentration may be between 0.5% and 20%. In another embodiment, the final protein concentration may be between 0.5% and 15%. In another embodiment, the final protein concentration may be between 0.5% and 10%. In another embodiment, the final protein concentration may be between 0.5% and 5%. In one embodiment, a composition with a final protein concentration as described above will be formulated for intravenous administration.
In certain embodiments, the final protein concentration may be between 5% and 25%. In another embodiment, the final protein concentration may be between 10% and 25%. In another embodiment, the final protein concentration may be between 15% and 25%. In another embodiment, the final protein concentration may be between 20% and 25%. In one embodiment, a composition with a final protein concentration as described above will be formulated for subcutaneous or intramuscular administration.
In certain embodiments, the labile therapeutic protein will be formulated at a final concentration of from 0.05 g/L to 250 g/L. In certain embodiments, the labile protein is formulated at a final concentration of 0.05±0.01 g/L, 0.06±0.01 g/L, 0.07±0.01 g/L, 0.08±0.01 g/L, 0.09±0.01 g/L, 0.1±0.01 g/L, 0.2±0.02 g/L, 0.3±0.03 g/L, 0.4±0.04 g/L, 0.5±0.05 g/L, 0.6±0.06 g/L, 0.7±0.07 g/L, 0.8±0.08 g/L, 0.9±0.09 g/L, 1±0.1 g/L, 210.2 g/L, 3±0.3 g/L, 4±0.4 g/L, 5±0.5 g/L, 6±0.6 g/L, 7±0.7 g/L, 8±0.8 g/L, 9±0.9 g/L, 10±1 g/L, 11±1.1 g/L, 12±1.2 g/L, 13±1.3 g/L, 14±1.4 g/L, 15±1.5 g/L, 16±1.6 g/L, 17±1.7 g/L, 18±1.8 g/L, 19±1.9 g/L, 20±2 g/L, 21±2.1 g/L, 22±2.2 g/L, 23±2.3 g/L, 24±2.4 g/L, 25±2.5 g/L, 26±2.6 g/L, 27±2.7 g/L, 28±2.8 g/L, 29±2.9 g/L, 30±3 g/L, 35±3.5 g/L, 40±4 g/L, 45±4.5 g/L, 50±5 g/L, 55±5.5 g/L, 60±6 g/L, 65±6.5 g/L, 70±7 g/L, 75±7.5 g/L, 80±8 g/L, 85±8.5 g/L, 90±9 g/L, 95±9.5 g/L, 100±10 g/L, 110±11 g/L, 120±12 g/L, 130±13 g/L, 140±14 g/L, 150±15 g/L, 160±16 g/L, 170±17 g/L, 180±18 g/L, 190±19 g/L, 200±20 g/L, 210±21 g/L, 220±22 g/L, 230±23 g/L, 240±24 g/L, 250±25 g/L, or higher concentrations, depending upon the characteristics of the protein being formulated, the intended therapeutic use of the protein, and the preferred method of administration. In yet other embodiments, the final concentration of the labile protein in the formulation is 0.05 g/L, 0.06 g/L, 0.07 g/L, 0.08 g/L, 0.09 g/L, 0.1 g/L, 0.2 g/L, 0.3 g/L, 0.4 g/L, 0.5 g/L, 0.6 g/L, 0.7 g/L, 0.8 g/L, 0.9 g/L, 1 g/L, 2 g/L, 3 g/L, 4 g/L, 5 g/L, 6 g/L, 7 g/L, 8 g/L, 9 g/L, 10 g/L, 11 g/L, 12 g/L, 13 g/L, 14 g/L, 15 g/L, 16 g/L, 17 g/L, 18 g/L, 19 g/L, 20 g/L, 21 g/L, 22 g/L, 23 g/L, 24 g/L, 25 g/L, 26 g/L, 27 g/L, 28 g/L, 29 g/L, 30 g/L, 35 g/L, 40 g/L, 45 g/L, 50 g/L, 55 g/L, 60 g/L, 65 g/L, 70 g/L, 75 g/L, 80 g/L, 85 g/L, 90 g/L, 95 g/L, 100 g/L, 110 g/L, 120 g/L, 130 g/L, 140 g/L, 150 g/L, 160 g/L, 170 g/L, 180 g/L, 190 g/L, 200 g/L, 210 g/L, 220 g/L, 230 g/L, 240 g/L, 250 g/L, or higher
In certain embodiments, the formulations provided herein will stabilize a labile therapeutic protein composition when stored at a temperature between 2° C. and 42° C. In one embodiments, a labile therapeutic protein will be stabilized by the formulations provided herein when stored under refrigeration, i.e., stored at a temperature between 2° C. and 8° C. In another embodiment, a labile therapeutic protein will be stabilized by the formulations provided herein when stored at room temperature, i.e., stored at a temperature between 20° C. and 25° C. In other embodiments, the protein may be stabilized when stored at a temperature between 28° C. and 32° C. In yet another embodiment, the protein may be stabilized when stored at a temperature between 38° C. and 42° C. The temperatures at which a labile therapeutic protein will be stabilized by the formulations provided herein will be dependent upon the characteristics of the individual protein, which can readily be determined by one of skill in the art.
Likewise, the extent of time for which a labile protein is stabilized by the formulations provided herein will depend upon the individual protein. For example, in one embodiment, a storage stable, aqueous composition provided herein will be stable for at least 2 months. In another embodiment, the composition will be stable for at least 3 months. In yet other embodiment, the composition will be stable for at least 1, 2, 3, 4, 5, 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, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or more months. In a preferred embodiment, the composition will be stable for at least 6 months. In a more preferred embodiment, the composition will be stable for at least 1 year. In another preferred embodiment, the composition will be stable for at least 2 years.
The extent to which a labile therapeutic protein is stabilized by the formulations provided herein may also be expressed as a percentage increase in the time the composition is stable under standard storage conditions. For example, in one embodiment, a labile therapeutic protein composition may be stable under storage conditions for at least 25% longer when formulated with an alkali metal chloride salt at a mildly acidic to neutral pH, as provided herein, as compared to the stability of the same protein under mildly acidic to neutral pH in the absence of the alkali metal chloride salt. In other embodiments, the composition may be stable for at least 50% longer when formulated according to the present invention, or at least 75%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, or longer when formulated according to the present invention. For example, in one embodiment, the labile therapeutic protein composition may be stable under storage conditions for from 25% to 1000% longer when formulated with an alkali metal chloride salt at a mildly acidic to neutral pH, as provided herein, as compared to the stability of the same protein under mildly acidic to neutral pH in the absence of the alkali metal chloride salt. In other embodiments, the composition may be stable for from 50% to 1000%, 100% to 1000%, 200% to 1000%, 300% to 1000%, 400% to 1000%, 500% to 1000%, 600% to 1000%, or 700% to 1000%, when formulated according to the present invention.
Protein stability may be measured using various metrics, including but not limited to, the extent or rate of protein aggregation, the loss of enzymatic activity, the loss of anti-antigen titer, and/or the extent or rate of protein degradation. One of skill in the art will recognize that certain metrics will be more or less relevant to individual proteins. For example, the stability of an enzyme may be determined by monitoring the loss of enzymatic activity over time, but not by monitoring the loss of an anti-antigen titer. Conversely, the stability of an antibody may be measured by monitoring the loss of anti-antigen titer, but not by enzymatic activity.
Accordingly, in one embodiment, a labile immunoglobulin composition is an immunoglobulin composition that losses from 20% to 100% of its anti-antigen titer when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 6 months. In another embodiment, a labile immunoglobulin composition is an immunoglobulin composition that losses from 30% to 100% of its anti-antigen titer when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 6 months. In another embodiment, a labile immunoglobulin composition is an immunoglobulin composition that losses from 40% to 100% of its anti-antigen titer when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 6 months. In another embodiment, a labile immunoglobulin composition is an immunoglobulin composition that losses from 50% to 100% of its anti-antigen titer when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 6 months.
In one embodiment, the stability of a labile protein composition may be determined by monitoring the extent or rate of protein aggregation within the formulation. Protein aggregation may be determined for example, by size exclusion chromatography (SEC), high performance size exclusion chromatography (HP-SEC), dynamic light scattering, non-denaturing gel electrophoresis and the like. Although the absolute aggregation level at which a labile protein composition will be considered unstable will vary from protein to protein, the level of aggregation that results in a significant loss of the therapeutic value of the composition will generally be regarded as unstable.
In another embodiment, wherein the labile therapeutic protein is an enzyme, the stability of the composition may be determined by monitoring the loss of bulk enzymatic activity in the preparation. For example, in one embodiment, a 20% loss of enzymatic activity will correspond to an unstable composition. In other embodiments, a 10% loss of enzymatic activity, or a 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or higher loss of enzymatic activity will correspond to an unstable composition.
In one embodiment, a labile protein composition is an enzyme composition that losses from 10% to 100% of enzymatic activity when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.5 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 1 month. In another embodiment, a labile protein composition is an enzyme composition that losses from 20% to 100% of enzymatic activity when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 1 month. In one embodiment, a labile protein composition is an enzyme composition that losses from 30% to 100% of enzymatic activity when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 1 month. In one embodiment, a labile protein composition is an enzyme composition that losses from 40% to 100% of enzymatic activity when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 1 month. In one embodiment, a labile protein composition is an enzyme composition that losses from 50% to 100% of enzymatic activity when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 1 month.
In one embodiment, a labile protein composition is an enzyme composition that losses from 10% to 100% of enzymatic activity when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.5 and stored at a particular temperature (e.g.; from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 2 months. In another embodiment, a labile protein composition is an enzyme composition that losses from 20% to 100% of enzymatic activity when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 2 months. In one embodiment, a labile protein composition is an enzyme composition that losses from 30% to 100% of enzymatic activity when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 2 months. In one embodiment, a labile protein composition is an enzyme composition that losses from 40% to 100% of enzymatic activity when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 2 months. In one embodiment, a labile protein composition is an enzyme composition that losses from 50% to 100% of enzymatic activity when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 2 months.
In one embodiment, a labile protein composition is an enzyme composition that losses from 10% to 100% of enzymatic activity when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.5 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 3 months. In another embodiment, a labile protein composition is an enzyme composition that losses from 20% to 100% of enzymatic activity when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 3 months. In one embodiment, a labile protein composition is an enzyme composition that losses from 30% to 100% of enzymatic activity when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 3 months. In one embodiment, a labile protein composition is an enzyme composition that losses from 40% to 100% of enzymatic activity when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 3 months. In one embodiment, a labile protein composition is an enzyme composition that losses from 50% to 100% of enzymatic activity when formulated with less than 75 mM (i.e., between 0 mM and 75 mM) of an alkali metal chloride salt at a pH from 5.5 to 7.0 and stored at a particular temperature (e.g., from 2° C. to 8° C., from 20° C. to 25° C., from 28° C. to 32° C., or from 38° C. to 42° C.) for between 1 day and 3 months.
In another embodiment, the stability of a labile protein composition may be determined by monitoring the extent or rate of protein degradation within the formulation. Protein degradation may be determined for example, by size exclusion chromatography (SEC), high performance size exclusion chromatography (HP-SEC), dynamic, light scattering, non-denaturing gel electrophoresis and the like. Although the absolute degradation level at which a labile protein composition will be considered unstable will vary from protein to protein, the level of degradation that results in a significant loss of the therapeutic value of the composition will generally be regarded as unstable.
Accordingly, in one aspect, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein, the composition comprising between 75 mM and 200 mM of an alkali metal chloride salt, a stabilizing agent, and a pH between 5.5 and 7.5. In a specific embodiment, the stabilizing agent is a buffer. In a more specific embodiment, the stabilizing agent is an amino acid. In a yet more specific embodiment, the amino acid is glycine, histidine, or proline. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein, the composition consisting essentially of: between 75 mM and 200 mM of an alkali metal chloride salt, a stabilizing agent, and a pH between 5.5 and 7.5. In a specific embodiment, the stabilizing agent is a buffer. In a more specific embodiment, the stabilizing agent is an amino acid. In a yet more specific embodiment, the amino acid is glycine, histidine, or proline. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein, the composition consisting of: between 75 mM and 200 mM of an alkali metal chloride salt, a stabilizing agent, and a pH between 5.5 and 7.5. In a specific embodiment, the stabilizing agent is a buffer. In a more specific embodiment, the stabilizing agent is an amino acid. In a yet more specific embodiment, the amino acid is glycine, histidine, or proline. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a closely related embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein, the composition comprising: between about 75 mM and about 200 mM of an alkali metal chloride salt, a stabilizing agent, and a pH between about 5.5 and about 7.5. In a specific embodiment, the stabilizing agent is a buffer. In a more specific embodiment, the stabilizing agent is an amino acid. In a yet more specific embodiment, the amino acid is glycine, histidine, or proline. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein, the composition consisting essentially of: between about 75 mM and about 200 mM of an alkali metal chloride salt, a stabilizing agent, and a pH between about 5.5 and about 7.5. In a specific embodiment, the stabilizing agent is a buffer. In a more specific embodiment, the stabilizing agent is an amino acid. In a yet more specific embodiment, the amino acid is glycine, histidine, or proline. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein, the composition consisting of: between about 75 mM and about 200 mM of an alkali metal chloride salt, a stabilizing agent, and a pH between about 5.5 and about 7.5. In a specific embodiment, the stabilizing agent is a buffer. In a more specific embodiment, the stabilizing agent is an amino acid. In a yet more specific embodiment, the amino acid is glycine, histidine, or proline. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
1. Alkali Metal Chloride Salts
Surprisingly, it was found that the addition of a moderate level of an alkali metal chloride salt to an aqueous composition of a labile therapeutic protein formulated at a mildly acidic to neutral pH significantly stabilized the labile protein, regardless of the identity of the protein. In one embodiment, a stabilizing formulation provided herein contains between 75 mM and 200 mM of an alkali metal chloride salt. In a preferred embodiment, the formulation contains between 100 mM and 200 mM of an alkali metal chloride salt. In another embodiment, the formulation contains between 150 mM and 200 mM of an alkali metal chloride salt. In another embodiment, the formulation contains between 75 mM and 175 mM of an alkali metal chloride salt. In another embodiment, the formulation contains between 75 mM and 150 mM of an alkali metal chloride salt. In another embodiment, the formulation contains between 75 mM and 125 mM of an alkali metal chloride salt. In another embodiment, the formulation contains between 100 mM and 175 mM of an alkali metal chloride salt. In another embodiment, the formulation contains between 100 mM and 150 mM of an alkali metal chloride salt. In another embodiment, the formulation contains between 100 mM and 125 mM of an alkali metal chloride salt. In another embodiment, the formulation contains between 125 mM and 200 mM of an alkali metal chloride salt. In another embodiment, the formulation contains between 125 mM and 175 mM of an alkali metal chloride salt. In another embodiment, the formulation contains between 125 mM and 150 mM of an alkali metal chloride salt. In yet other embodiments, the formulation contains 70±7 mM, 75±7.5 mM, 80±8 mM, 90±9 mM, 100 mM±10, 110±11 mM, 120 mM±12, 125±12.5 mM, 130±13 mM, 140±14 mM, 150±15 mM, 160±16 mM, 170±17 mM, 175±17.5 mM, 180±18 mM, 190±19 mM, 200±20 mM, 210±21 mM, or 220±22 mM of an alkali metal chloride salt. In yet other more specific embodiments, the formulation contains 70 mM, 75 mM, 80 mM, 85 mM, 90 mM, 95 mM, 100 mM, 105 mM, 110 mM, 115 mM, 120 mM, 125 mM, 130 mM, 135 mM, 140 mM, 145 mM, 150 mM, 155 mM, 160 mM, 165 mM, 170 mM, 175 mM, 180 mM, 185 mM, 190 mM, 195 mM, 200 mM, 205 mM, 210 mM, 215 mM, or 220 mM of an alkali metal chloride salt. In one preferred embodiment, the alkali metal chloride salt is sodium chloride. In another preferred embodiment, the alkali metal chloride salt is potassium chloride.
Accordingly, in one aspect, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; from 125 mM to 175 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; from 125 mM to 175 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; from 125 mM to 175 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In another embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; 150±15 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; 150±15 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; 150±15 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In another embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; a stabilizing agent; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; a stabilizing agent; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; a stabilizing agent; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride.
2. Stabilizing Agents
The pharmaceutical compositions provided herein will typically comprise one or more buffering agents or pH stabilizing agents suitable for intravenous, intravitreal, subcutaneous, and/or intramuscular administration. Non-limiting examples of buffering agents suitable for formulating the storage stable compositions provided herein include glycine, histidine, proline, or other amino acids, salts like citrate, phosphate, acetate, glutamate, tartrate, benzoate, lactate, gluconate, malate, succinate, formate, propionate, carbonate, or any combination thereof adjusted to an appropriate pH. Generally, the buffering agent will be sufficient to maintain a suitable pH in the formulation for an extended period of time.
In a preferred embodiment, the stabilizing agent employed in the storage stable, labile therapeutic protein formulations provided herein is an amino acid. Non-limiting examples of amino acids include, isoleucine, alanine, leucine, asparagine, lysine, aspartic acid, methionine, cysteine, phenylalanine, glutamic acid, threonine, glutamine, tryptophan, glycine, valine, proline, selenocysteine, serine, tyrosine, arginine, histidine, ornithine, taurine, combinations thereof, and the like. In one embodiment, the stabilizing amino acids include arginine, histidine, lysine, serine, proline, glycine, alanine, threonine, and a combination thereof. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In yet another preferred embodiment, the amino acid is histidine.
For purposes of further stabilizing the compositions provided herein, the amino acid will typically be added to the formulation at a concentration between 5 mM and 0.75 M. In one embodiment, at least 100 mM of the amino acid is added to the formulation. In another embodiment, at least 200 mM of the amino acid is added to the formulation. In yet another embodiment, at least 250 mM of the amino acid is added to the formulation. In yet other embodiments, the formulations provided herein will contain at least 25 mM, 50 mM, 75 mM, 100 mM, 150 mM, 200 mM, 250 mM, 300 mM, 350 mM, 400 mM, 450 mM, 500 mM, 550 mM, 600 mM, 650 mM, 700 mM, 750 mM, or more of the amino acid.
In one embodiment, the concentration of buffering agent in the formulation will be at or about between 5 mM and 500 mM. In certain embodiments, the concentration of the buffering agent in the formulation will be at or about 5 mM, 10 mM, 15 mM, 20 mM, 25 mM, 50 mM, 75 mM, 100 mM, 125 mM, 150 mM, 175 mM, 200 mM, 225 mM, 250 mM, 275 mM, 300 mM, 325 mM, 350 mM, 375 mM, 400 mM, 425 mM, 450 mM, 475 mM, 500 mM or higher.
Accordingly, in one aspect, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In another embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; an amino acid; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; an amino acid; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; an amino acid; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In another aspect, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM glycine; and a pH of from 5.5 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM glycine; and a pH of from 5.5 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM glycine; and a pH of from 5.5 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In another embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; from 5 mM to 500 mM glycine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; from 5 mM to 500 mM glycine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; from 5 mM to 500 mM glycine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In another aspect, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM proline; and a pH of from 5.5 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM proline; and a pH of from 5.5 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM proline; and a pH of from 5.5 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In another embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; from 5 mM to 500 mM proline; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; from 5 mM to 500 mM proline; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; from 5 mM to 500 mM proline; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In another aspect, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM histidine; and a pH of from 5.5 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM histidine; and a pH of from 5.5 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM histidine; and a pH of from 5.5 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In another embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; from 5 mM to 500 mM histidine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; from 5 mM to 500 mM histidine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; from 5 mM to 500 mM histidine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
3. Excipients
In certain embodiments, the storage stable labile therapeutic protein aqueous compositions provided herein further comprise one or more excipients. Non-limiting examples of excipients that can be included in the formulations provided herein include non-ionic surfactants, bulking agents (e.g., sugars and sugar alcohols), antioxidants, polysaccharides, and pharmaceutically acceptable water-soluble polymers (e.g., poly(acrylic acid), poly(ethylene oxide), poly(ethylene glycol), poly(vinyl pyrrolidone), hydroxyethyl cellulose, hydroxypropyl cellulose, and starch).
In one embodiment, the excipient is an agent for adjusting the osmolarity of the composition. Non-limiting examples of osmolarity agents include mannitol, sorbitol, glycerol, sucrose, glucose, dextrose, levulose, fructose, lactose, trehalose, polyethylene glycols, phosphates, calcium chloride, calcium gluconoglucoheptonate, dimethyl sulfone, and the like.
Accordingly, in one aspect, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; an antioxidant; and a pH of from 5.5 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; an antioxidant, and a pH of from 5.5 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; an antioxidant; and a pH of from 5.5 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In another embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; an amino acid; an antioxidant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; an amino acid; an antioxidant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment; the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; an amino acid; an antioxidant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
Accordingly, in one aspect, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; a sugar and/or sugar alcohol; and a pH of from 5.5 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; a sugar and/or sugar alcohol, and a pH of from 5.5 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; a sugar and/or sugar alcohol; and a pH of from 5.5 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In another embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: a labile therapeutic protein; an amino acid; a sugar and/or sugar alcohol; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; an amino acid; a sugar and/or sugar alcohol; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; an amino acid; a sugar and/or sugar alcohol; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In one embodiment, the pH of the formulation is between 5.5 and 7.0. In a specific embodiment, the pH of the formulation is between 5.5 and 6.5. In yet another embodiment, the pH of the formulation is between 6.0 and 7.0.
4. Administration
Formulations of the storage stable composition provided herein are delivered to the individual by any pharmaceutically suitable means of administration. Various delivery systems are known and can be used to administer the composition by any convenient route. In one embodiment the compositions of the invention are administered systemically. For systemic use, the composition is formulated for parenteral (e.g. intradermal, subcutaneous, transdermal implant, intracavernous, intravitreal, transscleral, intracerebral, intrathecal, epidural, intravenous, intracardiac, intramuscular, intraosseous, intraperitoneal, and nanocell injection) or enteral (e.g., oral, vaginal or rectal) delivery according to conventional methods. The formulations can be administered continuously by infusion or by bolus injection. Some formulations encompass slow release systems. Preferred routes of administration will depend upon the indication being treated, managed, or prevented.
Single or multiple administrations of the compositions are carried out with the dose levels and pattern being selected by the treating physician. For the prevention or treatment of disease, the appropriate dosage depends on the type of disease to be treated, the severity and course of the disease, whether drug is administered for preventive or therapeutic purposes, previous therapy, the patient's clinical history and response to the therapeutic protein, and the discretion of the attending physician.
In a preferred embodiment, the storage stable compositions provided herein are formulated for parenteral administration including, but not limited to, intradermal, subcutaneous, transdermal implant, intracavernous, intravitreal, transscleral, intracerebral, intrathecal, epidural, intravenous, intracardiac, intramuscular, intraosseous, intraperitoneal, and nanocell injection administration. In one preferred embodiment, the compositions provided herein will be formulated for intravenous administration. In another preferred embodiment, the compositions provided herein will be formulated for subcutaneous administration. In yet another preferred embodiment, the compositions provided herein will be formulated for intramuscular administration.
In one aspect, the present invention provides storage stable, aqueous immunoglobulin compositions formulated at mildly acidic to neutral pH with a moderate concentration of a metal chloride salt and a stabilizing agent.
Any immunoglobulin may be stabilized by the formulations provided herein. Non-limiting examples of immunoglobulin preparations that may be stabilized include, plasma-derived immunoglobulin preparations, recombinant polyclonal or monoclonal preparations, minibodies, diabodies, triabodies, antibody fragments such as Fv, Fab and F(ab)2 or fragmented antibodies such as monovalent or multivalent single chain Fvs (scFv, sc(Fv)2, minibodies, diabodies, and triabodies such as scFv dimers) in which the variable regions of an antibody are joined together via a linker such as a peptide linker, and the like. Recombinant antibodies include murine antibodies, rodent antibodies, human antibodies, chimeric human antibodies (e.g., human/murine chimeras), humanized antibodies (e.g., humanized murine antibodies), and the like. In preferred embodiments, the recombinant antibody is a human, chimeric human, or humanized antibody suitable for administration to a human. In a preferred embodiment, the immunoglobulin in a full length, or near full length immunoglobulin, which will generally be more labile then engineered fragments thereof.
Generally, storage stable immunoglobulin formulations provided herein will be stabilized at room temperature (i.e., between 20° C. and 25° C.) for an extended period of time. For example, in one embodiment, a storage stable, aqueous immunoglobulin composition will be stable when stored at room temperature for at least about 2 months. In another embodiment, the composition will be stable for at least about 3 months. In yet other embodiment, the composition will be stable for at least 1 about month, or at least about 2, 3, 4, 5, 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, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or more months. In a preferred embodiment, the composition will be stable for at least about 6 months. In a more preferred embodiment, the composition will be stable for at least about 1 year. In a more preferred embodiment, the composition will be stable for at least about 2 years.
In one embodiment, the storage stable, aqueous immunoglobulin composition will be stable for at least six months at a temperature between about 28° C. and about 32° C. In a specific embodiment, the storage stable, aqueous immunoglobulin composition will be stable for at least one year at a temperature between about 28° C. and about 32° C. In a more specific embodiment, the storage stable, aqueous immunoglobulin composition will be stable for at least two years at a temperature between about 28° C. and about 32° C. In another embodiment, the storage stable, aqueous immunoglobulin composition will be stable for at least one month at a temperature between about 38° C. and about 42° C. In a specific embodiment, the storage stable, aqueous immunoglobulin composition will be stable for at least three months at a temperature between about 38° C. and about 42° C. In a more specific embodiment, the storage stable, aqueous immunoglobulin composition will be stable for at least one year at a temperature between about 38° C. and about 42° C.
In one embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 10%. In a preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 9%. In a more preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 8%. In a more preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 7%. In a more preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 6%. In a more preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 5%. In a more preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 4%. In a more preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 3%. In a most preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 2%.
In one embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 10% and the percentage of immunoglobulin in the monomeric state is no less than 85%. In a preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 9% and the percentage of immunoglobulin in the monomeric state is no less than 85%. In a preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 8% and the percentage of immunoglobulin in the monomeric state is no less than 85%. In a preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 7% and the percentage of immunoglobulin in the monomeric state is no less than 85%. In a preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 6% and the percentage of immunoglobulin in the monomeric state is no less than 85%. In a preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 5% and the percentage of immunoglobulin in the monomeric state is no less than 85%. In a preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 4% and the percentage of immunoglobulin in the monomeric state is no less than 85%. In a preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 3% and the percentage of immunoglobulin in the monomeric state is no less than 85%. In a most preferred embodiment, the composition is considered stable as long as the percentage of immunoglobulin in the aggregated state no more than 2% and the percentage of immunoglobulin in the monomeric state is no less than 85%.
In one embodiment, wherein the labile therapeutic protein is an antibody or fragment thereof, the stability of the composition may be determined by monitoring the loss of anti-antigen titer. The level of anti-antigen titer may be determined, for example, by an immunoassay. A variety of immunoassay formats may be used for this purpose. For example, solid-phase ELISA immunoassays are routinely used to determine antigen titer (see, e.g., Harlow & Lane, Using Antibodies, A Laboratory Manual (1998) for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity). In one embodiment, a 20% loss of anti-antigen titer will correspond to an unstable composition. In other embodiments, a 10% loss of anti-antigen titer, or a 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or higher loss of anti-antigen titer will correspond to an unstable composition.
After formulation, the storage stable, aqueous immunoglobulin compositions provided herein are typically sterile filtered and dispensed into a sterile containment vessel, which is sealed air-tight, for example, using a rubber stopper. Immunoglobulin compositions in the air-tight vessels are preferably protected from ambient light by storage in a dark place, the use of a tinted vessel material (typically glass or plastic), and/or covering the surface of the vessel with an opaque substance.
In certain embodiments, the headspace air in the containment vessel is replaced with an inert gas. The inert gas helps to maintain an inert atmosphere above the liquid composition. In one embodiment, the liquid is overlaid with inert gas. In another embodiment the liquid is degassed before overlaying it with inert gas, meaning that residual oxygen in the atmosphere may vary. In the context of the present invention, when an immunoglobulin composition is stored in a vessel in which the headspace air has been replaced with an inert gas, the composition has been overlaid with inert gas, or the composition is degassed prior to overlaying with inert gas, the composition is said to be “stored under inert gas.” Non-limiting examples of inert gasses than may be used in conjunction with the present invention include, nitrogen, argon, carbon dioxide, helium, krypton, and xenon. In one particular embodiment, the inert gas is nitrogen. In another particular embodiment, the inert gas is argon.
1. Plasma-Derived Immunoglobulins
Preparations of concentrated immunoglobulins (especially IgG) isolated from pooled human plasma are used for treating a variety of medical conditions, including immune deficiencies, inflammatory and autoimmune diseases, and acute infections. One IgG product, intravenous immunoglobulin or IVIG, is formulated for intravenous administration, for example, at a concentration of at or about 10% IgG. Concentrated immunoglobulins may also be formulated for subcutaneous or intramuscular administration, for example, at a concentration at or about 20% IgG.
Generally, plasma-derived immunoglobulin preparations formulated according to the present invention can be prepared from any suitable starting materials, for example, recovered plasma or source plasma. In a typical example, blood or plasma is collected from healthy donors. Immunoglobulins are isolated from the blood or plasma by suitable procedures, such as, for example, precipitation (alcohol fractionation or polyethylene glycol fractionation), chromatographic methods (ion exchange chromatography, affinity chromatography, immunoaffinity chromatography, etc.) ultracentrifugation, and electrophoretic preparation, and the like. (See, e.g., Cohn et al., J. Am. Chem. Soc. 68:459-75 (1946); Oncley et al., J. Am. Chem. Soc. 71:541-50 (1949); Barandun et al., Vox Sang. 7:157-74 (1962); Koblet et al., Vox Sang. 13:93-102 (1967); U.S. Pat. Nos. 5,122,373 and 5,177,194; PCT/US10/36470; and WO 2010/138736 the disclosures of which are hereby incorporated by reference in their entireties for all purposes).
In many cases, immunoglobulins are prepared from gamma globulin-containing compositions produced by alcohol fractionation and/or ion exchange and affinity chromatography methods well known to those skilled in the art. For example, purified Cohn Fraction II is commonly used as a starting point for the further purification of immunoglobulins. The starting Cohn Fraction II paste is typically about 95 percent IgG and is comprised of the four IgG subtypes. The different subtypes are present in Fraction II in approximately the same ratio as they are found in the pooled human plasma from which they are obtained. The Fraction II is further purified before formulation into an administrable product. For example, the Fraction II paste can be dissolved in a cold purified aqueous alcohol solution and impurities removed via precipitation and filtration. Following the final filtration, the immunoglobulin suspension can be dialyzed or diafiltered (e.g., using ultrafiltration membranes having a nominal molecular weight limit of less than or equal to 100,000 daltons) to remove the alcohol. The solution can be concentrated or diluted to obtain the desired protein concentration and can be further purified by techniques well known to those skilled in the art.
Furthermore, additional preparative steps can be used to enrich a particular isotype or subtype of immunoglobulin. For example, protein A, protein G or protein H sepharose chromatography can be used to enrich a mixture of immunoglobulins for IgG, or for specific IgG subtypes. See generally, Harlow and Lane, Using Antibodies, Cold Spring Harbor Laboratory Press (1999); Harlow and Lane, Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory Press (1988); and U.S. Pat. No. 5,180,810, the disclosures of which are hereby incorporated by reference in their entireties for all purposes.
As will be recognized by one of skill in the art, immunoglobulin compositions isolated from pooled plasma contain impurities carried over from the starting plasma. Typically, pharmaceutically acceptable plasma-derived immunoglobulin compositions will contain at least 90% immunoglobulins, preferably at least 95% immunoglobulins, more preferably at least 98% immunoglobulins; most preferably at least 99% immunoglobulins, expressed as a function of total protein content. For example, GAMMAGARD® LIQUID (Baxter International; Deerfield, Ill.) is a plasma-derived immunoglobulin composition formulated at 100 g/L protein. According to the specifications, at least 98% of the protein is immune globulin, the average immunoglobulin A (IgA) concentration is 37 μg/mL, and immunoglobulin M is present in trace amounts (GAMMAGARD® LIQUID Prescribing Information). Accordingly, unless otherwise specified, an immunoglobulin composition provided herein comprising; consisting essentially of; or consisting of “a plasma-derived immunoglobulin” may contain up to 10% plasma protein impurities carried through during the manufacturing process.
In a particular embodiment, the immunoglobulin composition isolated from pooled plasma comprises at least 90% IgG immunoglobulins. In a specific embodiment, the immunoglobulin composition isolated from pooled plasma comprises at least 95% IgG immunoglobulins. In a more specific embodiment, the immunoglobulin composition isolated from pooled plasma comprises at least 98% IgG immunoglobulins. In a yet more specific embodiment, the immunoglobulin composition isolated from pooled plasma comprises at least 99% IgG immunoglobulins. In certain embodiments, the IgG immunoglobulin composition isolated from pooled plasma further comprises IgA and/or IgM immunoglobulins.
In another embodiment, the immunoglobulin composition isolated from pooled plasma comprises at least 10% IgA. In a specific embodiment, the immunoglobulin composition isolated from pooled plasma comprises at least 25% IgA. In a more specific embodiment, immunoglobulin composition isolated from pooled plasma comprises at least 50% IgA. In yet other embodiments, the immunoglobulin composition isolated from pooled plasma comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more IgA. In certain embodiments, the IgA immunoglobulin composition isolated from pooled plasma further comprises IgG and/or IgM immunoglobulins.
In another embodiment, the immunoglobulin composition isolated from pooled plasma comprises at least 10% IgM. In a specific embodiment, the immunoglobulin composition isolated from pooled plasma comprises at least 25% IgM. In a more specific embodiment, immunoglobulin composition isolated from pooled plasma comprises at least 50% IgM. In yet other embodiments, the immunoglobulin composition isolated from pooled plasma comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more IgM. In certain embodiments, the IgM immunoglobulin composition isolated from pooled plasma further comprises IgG and/or IgA immunoglobulins.
Accordingly, in one aspect, the present invention provides a storage stable, aqueous composition comprising: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.0. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.0. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.0. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a plasma-derived immunoglobulin; from 125 mM to 175 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.0. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a plasma-derived immunoglobulin; from 125 mM to 175 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.0. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a plasma-derived immunoglobulin; from 125 mM to 175 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.0. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a plasma-derived immunoglobulin; a stabilizing agent; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; a stabilizing agent; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; a stabilizing agent; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In one embodiment, the present invention provides a storage stable, aqueous composition comprising: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a plasma-derived immunoglobulin; an amino acid; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a plasma-derived immunoglobulin; an amino acid; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a plasma-derived immunoglobulin; an amino acid; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In another aspect, the present invention provides a storage stable, aqueous composition comprising: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM glycine; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM glycine; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM glycine; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a plasma-derived immunoglobulin; from 5 mM to 500 mM glycine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a plasma-derived immunoglobulin; from 5 mM to 500 mM glycine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a plasma-derived immunoglobulin; from 5 mM to 500 mM glycine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In another aspect, the present invention provides a storage stable, aqueous composition comprising: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM proline; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM proline; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM proline; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a plasma-derived immunoglobulin; from 5 mM to 500 mM proline; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a plasma-derived immunoglobulin; from 5 mM to 500 mM proline; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a plasma-derived immunoglobulin; from 5 mM to 500 mM proline; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In another aspect, the present invention provides a storage stable, aqueous composition comprising: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM histidine; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM histidine; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM histidine; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a plasma-derived immunoglobulin; from 5 mM to 500 mM histidine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a plasma-derived immunoglobulin; from 5 mM to 500 mM histidine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a plasma-derived immunoglobulin; from 5 mM to 500 mM histidine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In one aspect, the present invention provides a storage stable, aqueous composition comprising: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; an antioxidant; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; an antioxidant; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; an antioxidant; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a plasma-derived immunoglobulin; an amino acid; an antioxidant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a plasma-derived immunoglobulin; an amino acid; an antioxidant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a plasma-derived immunoglobulin; an amino acid; an antioxidant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In one aspect, the present invention provides a storage stable, aqueous composition comprising: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; a sugar and/or sugar alcohol; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; a sugar and/or sugar alcohol; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a plasma-derived immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; a sugar and/or sugar alcohol; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a plasma-derived immunoglobulin; an amino acid; a sugar and/or sugar alcohol; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a plasma-derived immunoglobulin; an amino acid; a sugar and/or sugar alcohol; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a plasma-derived immunoglobulin; an amino acid; a sugar and/or sugar alcohol; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the plasma-derived immunoglobulin comprises IgG.
a. Hyper-Immune Immunoglobulins
In a specific embodiment, the storage stable, plasma derived immunoglobulin composition is a hyper-immune immunoglobulin preparation. For example, in certain embodiments, the hyper-immune preparation may be an anti-tetanus, anti-D, anti-varicella, anti-rabies, anti-CMV, anti-hepatitis A, or anti-hepatitis B immunoglobulin preparation.
As demonstrated in Example 3, plasma derived anti-tetanus and anti-D preparations are stabilized by the addition of between about 100 mM and about 200 mM of an alkali metal chloride salt (e.g., sodium chloride) at a pH between about 5.5 and about 6.5. As shown in
Accordingly, in one embodiment, the present invention provides a storage stable, plasma derived hyper-immune immunoglobulin aqueous composition comprising between about 75 mM and about 200 mM of an alkali metal chloride salt, a stabilizing agent, and a pH between about 5.5 and about 6.5. In a preferred embodiment, the composition comprises between about 100 mM and about 200 mM of an alkali metal chloride salt, a stabilizing agent, and a pH between about 5.5 and about 6.5. In another preferred embodiment, the composition comprises between about 100 mM and about 200 mM of an alkali metal chloride salt, a stabilizing agent, and a pH between about 5.5 and about 6.0. In a preferred embodiment, the salt is sodium chloride. In another preferred embodiment, the salt is potassium chloride.
In one embodiment, the storage stable, plasma derived hyper-immune immunoglobulin aqueous compositions provided herein have a protein concentration of between about 30 g/L and about 250 g/L. In certain embodiments, the protein concentration of the hyper-immune immunoglobulin composition is between about 50 g/L and about 200 g/L, or between about 70 g/L and about 150 g/L, or between about 90 g/L and about 120 g/L, or any suitable concentration within these ranges, for example about 30 g/L, or about 35 g/L, 40 g/L, 45 g/L, 50 g/L, 55 g/L, 60 g/L, 65 g/L, 70 g/L, 75 g/L, 80 g/L, 85 g/L, 90 g/L, 95 g/L, 100 g/L, 105 g/L, 110 g/L, 115 g/L, 120 g/L, 125 g/L, 130 g/L, 135 g/L, 140 g/L, 145 g/L, 150 g/L, 155 g/L, 160 g/L, 165 g/L, 170 g/L, 175 g/L, 180 g/L, 185 g/L, 190 g/L, 195 g/L, 200 g/L, 205 g/L, 210 g/L, 215 g/L, 220 g/L, 225 g/L, 230 g/L, 235 g/L, 240 g/L, 245 g/L, 250 g/L, or higher. In a preferred embodiment, the aqueous hyper-immune immunoglobulin composition will have a concentration of between about 100 g/L and about 170 g/L.
The storage stable plasma derived hyper-immune immunoglobulin aqueous compositions provided herein will be stabilized at room temperature for an extended period of time. For example, in one embodiment, the storage stable, aqueous hyper-immune immunoglobulin composition will be stable for at least about 2 months. In another embodiment, the composition will be stable for at least about 3 months. In yet other embodiment, the composition will be stable for at least 1 about month, or at least about 2, 3, 4, 5, 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, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or more months. In a preferred embodiment, the composition will be stable for at least about 6 months. In a more preferred embodiment, the composition will be stable for at least about 1 year. In a more preferred embodiment, the composition will be stable for at least about 2 years.
2. Recombinant Immunoglobulins
In one aspect, the present invention provides storage stable, recombinant immunoglobulin preparations. Methods for obtaining recombinant antibodies, such as recombinant human antibodies are well known in the art. For example, a desired human antibody having a binding activity for a desired antigen can be obtained by in vitro immunizing human lymphocytes with the desired antigen or a cell expressing the desired antigen and fusing the immunized lymphocytes to human myeloma cells. A desired human antibody can also be obtained by immunizing a transgenic animal having all human antibody gene repertoires with an antigen (see, International Publications Nos. WO 93/12227, WO 92/03918, WO 94/02602, WO 94/25585, WO 96/34096, WO 96/33735). Methods for obtaining a human antibody by panning using a human antibody library are also known. For example, phages binding to an antigen can be selected by expressing the variable regions of a human antibody as single chain antibody fragments (scFv) on phage surfaces by a phage display method. The DNA sequences encoding the variable regions of the human antibody binding to the antigen can be determined by analyzing the genes of the selected phages. A whole human antibody can be obtained by preparing a suitable expression vector containing the determined DNA sequences of the scFv fragments binding to the antigen. These methods have already been well known from WO 92/01047, WO 92/20791, WO 93/06213, WO 93/11236, WO 93/19172, WO 95/01438, and WO 95/15388.
Methods for the expression of recombinant immunoglobulins are also well known in the art. For example, recombinant antibodies can be expressed in tissue or cell culture after transforming a recombinant gene for the construct into a suitable host. Suitable eukaryotic cells for use as hosts include animal, plant and fungal cells. Known animal cells include (1) mammalian cells such as CHO, COS, myeloma, BHK (baby hamster kidney), HeLa and Vero cells; (2) amphibian cells such as Xenopus oocytes; or (3) insect sells such as sf9, sf21 and Tn5. Known plant cells include cells of Nicotiana such as Nicotiana tabacum, which can be used as callus cultures. Known fungi include yeasts such as Saccharomyces spp., e.g. Saccharomyces serevisiae and filamentous fungi such as Aspergillus spp., e.g. Aspergillus niger. Prokaryotic cells can be used as producing systems using bacterial cells. Known bacterial cells include E. coli and Bacillus subtilis. Antibodies can be obtained by transforming these cells with an antibody gene of interest and culturing the transformed cells in vitro.
In one embodiment of the present invention, the media used to express a recombinant protein can be animal protein-free and chemically defined. Methods of preparing animal protein-free and chemically defined culture mediums are known in the art, for example in US 2008/0009040 and US 2007/0212770, which are both incorporated herein for all purposes. “Protein free” and related terms refers to protein that is from a source exogenous to or other than the cells in the culture, which naturally shed proteins during growth. In another embodiment, the culture medium is polypeptide free. In another embodiment, the culture medium is serum free. In another embodiment the culture medium is animal protein free. In another embodiment the culture medium is animal component free. In another embodiment, the culture medium contains protein, e.g., animal protein from serum such as fetal calf serum. In another embodiment, the culture has recombinant proteins exogenously added. In another embodiment, the proteins are from a certified pathogen free animal. The term “chemically defined” as used herein shall mean, that the medium does not comprise any undefined supplements, such as, for example, extracts of animal components, organs, glands, plants, or yeast. Accordingly, each component of a chemically defined medium is accurately defined. In a preferred embodiment, the media are animal-component free and protein free.
Typically a recombinant antibody formulated as provided herein is specific for a polypeptide associated with a disease or disorder. Non-limiting examples of such polypeptides include macrophage migration inhibitory factor (MIF), tissue factor pathway inhibitor (TFPI); alpha-1-antitrypsin; insulin A-chain; insulin B-chain; proinsulin; follicle stimulating hormone; calcitonin; luteinizing hormone; glucagon; clotting factors such as Factor II (prothrombin), Factor III (platelet tissue factor), Factor V, Factor VII, Factor VIII, Factor IX, Factor X, Factor XI, Factor XII, Factor XIII, and von Willebrand factor; anti-clotting factors such as Antithrombin III (ATIII), Protein C; atrial natriuretic factor; lung surfactant; a plasminogen activator, such as urokinase or human urine or tissue-type plasminogen activator (t-PA); bombesin; thrombin; hemopoietic growth factor; tumor necrosis factor-alpha and -beta; enkephalinase; RANTES (regulated on activation normally T-cell expressed and secreted); human macrophage inflammatory protein (MIP-1-alpha); a serum albumin such as human serum albumin; Muellerian-inhibiting substance; relaxin A-chain; relaxin B-chain; prorelaxin; mouse gonadotropin-associated peptide; a microbial protein, such as beta-lactamase; DNase; IgE; a cytotoxic T-lymphocyte associated antigen (CTLA), such as CTLA-4; inhibin; activin; vascular endothelial growth factor (VEGF); receptors for hormones or growth factors; protein A or D; rheumatoid factors; a neurotrophic factor such as bone-derived neurotrophic factor (BDNF), neurotrophin-3, -4, -5, or -6 (NT-3, NT-4, NT-5, or NT-6), or a nerve growth factor such as NGF-b; platelet-derived growth factor (PDGF); fibroblast growth factor such as aFGF and bFGF; epidermal growth factor (EGF); transforming growth factor (TGF) such as TGF-alpha and TGF-beta, including TGF-b1, TGF-b2, TGF-b3, TGF-b4, or TGF-b5; a tumor necrosis factor (TNF) such as TNF-alpha or TNF-beta; insulin-like growth factor-I and -II (IGF-I and IGF-II); des(1-3)-IGF-I (brain IGF-I), insulin-like growth factor binding proteins; CD proteins such as CD3, CD4, CD8, CD19, CD20, CD22 and CD40; erythropoietin; osteoinductive factors; immunotoxins; a bone morphogenetic protein (BMP); an interferon such as interferon-alpha, -beta, and -gamma; colony stimulating factors (CSFs), e.g., M-CSF, GM-CSF, and G-CSF; interleukins (ILs), e.g., IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9 and IL-10; superoxide dismutase; T-cell receptors; surface membrane proteins; decay accelerating factor; viral antigen such as, for example, a portion of the AIDS envelope; transport proteins; homing receptors; addressins; regulatory proteins; integrins such as CD11a, CD11b, CD11c, CD18, an ICAM, VLA-4 and VCAM; a tumor associated antigen such as HER2, HER3 or HER4 receptor; and fragments of any of the above-listed polypeptides.
Accordingly, in one aspect, the present invention provides a storage stable, aqueous composition comprising: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.0. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.0. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.0. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a recombinant immunoglobulin; from 125 mM to 175 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.0. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a recombinant immunoglobulin; from 125 mM to 175 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.0. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a recombinant immunoglobulin; from 125 mM to 175 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 5.5 to 7.0. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a recombinant immunoglobulin; a stabilizing agent; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a more specific embodiment, the stabilizing agent is an amino acid.
In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; a stabilizing agent; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; a stabilizing agent; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In one embodiment, the present invention provides a storage stable, aqueous composition comprising: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a recombinant immunoglobulin; an amino acid; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a recombinant immunoglobulin; an amino acid; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal, chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a recombinant immunoglobulin; an amino acid; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In another aspect, the present invention provides a storage stable, aqueous composition comprising: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM glycine; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM glycine; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM glycine; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a recombinant immunoglobulin; from 5 mM to 500 mM glycine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a recombinant immunoglobulin; from 5 mM to 500 mM glycine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a recombinant immunoglobulin; from 5 mM to 500 mM glycine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In another aspect, the present invention provides a storage stable, aqueous composition comprising: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM proline; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM proline; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM proline; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a recombinant immunoglobulin; from 5 mM to 500 mM proline; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a recombinant immunoglobulin; from 5 mM to 500 mM proline; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a recombinant immunoglobulin; from 5 mM to 500 mM proline; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In another aspect, the present invention provides a storage stable, aqueous composition comprising: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM histidine; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM histidine; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM histidine; and a pH of from 5.5 to 7.0. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a recombinant immunoglobulin; from 5 mM to 500 mM histidine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a recombinant immunoglobulin; from 5 mM to 500 mM histidine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a recombinant immunoglobulin; from 5 mM to 500 mM histidine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In one aspect, the present invention provides a storage stable, aqueous composition comprising: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; an antioxidant; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; an antioxidant; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; an antioxidant; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a recombinant immunoglobulin; an amino acid; an antioxidant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a recombinant immunoglobulin; an amino acid; an antioxidant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a recombinant immunoglobulin; an amino acid; an antioxidant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In one aspect, the present invention provides a storage stable, aqueous composition comprising: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; a sugar and/or sugar alcohol; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; a sugar and/or sugar alcohol; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a recombinant immunoglobulin; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; a sugar and/or sugar alcohol; and a pH of from 5.5 to 7.0. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a recombinant immunoglobulin; an amino acid; a sugar and/or sugar alcohol; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a recombinant immunoglobulin; an amino acid; a sugar and/or sugar alcohol; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a recombinant immunoglobulin; an amino acid; a sugar and/or sugar alcohol; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.0 and 7.0. In a specific embodiment, the recombinant immunoglobulin is a monoclonal antibody. In a more specific embodiment, the recombinant antibody is an anti-MIF antibody.
Surprisingly, it was found that the stabilizing effects of alkali metal chloride salts (e.g., sodium and potassium chloride) are applicable to a wide range of labile therapeutic proteins ranging from plasma derived immunoglobulin preparations to recombinant coagulation factors. Accordingly, in one aspect, the present invention provides storage stable, aqueous compositions of labile coagulation factors formulated at mildly acidic to neutral pH with a moderate concentration of a metal chloride salt and a stabilizing agent.
In certain embodiments, the labile coagulation factor is a plasma derived protein or preparation. In other embodiments, the labile coagulation protein is a recombinantly expressed coagulation protein. Methods for manufacturing recombinant and plasma derived coagulation factors are well known in the art. Non-limiting examples of labile coagulation factors that may be formulated according to the methods provided herein include, Factor II (prothrombin), Factor III (platelet tissue factor), Factor V, Factor VII, Factor VIII, Factor IX, Factor X, Factor XI, Factor XII, Factor XIII, von Willebrand Factor (vWF), and the like. In a preferred embodiment, the labile coagulation protein is selected from Factor VII, Factor VIII, Factor IX, and von Willebrand Factor (vWF). In another preferred embodiment, the labile coagulation protein is a vitamin-K dependent protein complex, for example, comprising Factor II, Factor IX, and Factor X, or comprising Factor II, Factor VII, Factor IX, and Factor X.
In one embodiment, the labile therapeutic coagulation protein is isolated from pooled plasma, i.e., plasma-derived coagulation factors. Methods for the isolation of many different coagulation factors are well known in the art. For example, Furuya et al. (“Implementation of a 20-nm pore-size filter in the plasma-derived factor VIII manufacturing process” Vox Sang. 2006 August; 91(2):119-25) describe a method for the purification of a virally reduced plasma-derived Factor VIII composition from pooled human plasma. Similarly, Kisiel et al. (“Activation of Bovine Factor VII (Proconvertin) by Factor XIIa (Activated Hageman Factor)” Biochemistry (1977) 16 (9):4189-4193) and Broze and Majerus (“Purification and Properties of Human Coagulation Factor VII” J Biol Chem. 1980 Feb. 25; 255(4):1242-7) describe methods for the purification of plasma-derived Factor VII. Methods for the purification of plasma-derived Protein K dependent coagulation complexes (e.g., Prothromplex, FEIBA) are described, for example, in U.S. Pat. Nos. 5,409,990 and 5,281,661. Finally, among other teachings, U.S. Pat. No. 4,786,726 and PCT Publication No. WO 2007/046631, describe purification of plasma-derived Factor FIX.
In other embodiments, the labile therapeutic coagulation protein is expressed recombinantly, i.e., recombinant coagulation factors. Methods for the expression and purification of many different coagulation factors are well known in the art. For example, U.S. Pat. Nos. 5,470,954; 6,100,061; 6,475,725; 6,555,391; 6,936,441; 7,094,574; 7,253,262; 6,919,311; 7,544,660; and 7,381,796, all of which are hereby incorporated by reference in their entireties for all purposes, describe the expression and purification of recombinant Factor VIII. Methods for the manufacture of recombinant Factor IX are also well known in the art and are described in, for example, U.S. Patent Application Publication 2008/207879, U.S. Pat. Nos. 4,770,999, and 5,521,070. Likewise, methods for the manufacture of recombinant Factor VII are described in, for example, U.S. Patent Application Publication Nos. 2010/120093 and 2009/047723.
In one embodiment of the present invention, the media used to express a recombinant coagulation factor can be animal protein-free and chemically defined. Methods of preparing animal protein-free and chemically defined culture mediums are known in the art, for example in US 2008/0009040 and US 2007/0212770, which are both incorporated herein for all purposes. “Protein free” and related terms refers to protein that is from a source exogenous to or other than the cells in the culture, which naturally shed proteins during growth. In another embodiment, the culture medium is polypeptide free. In another embodiment, the culture medium is serum free. In another embodiment the culture medium is animal protein free. In another embodiment the culture medium is animal component free. In another embodiment, the culture medium contains protein, e.g., animal protein from serum such as fetal calf serum. In another embodiment, the culture has recombinant proteins exogenously added. In another embodiment, the proteins are from a certified pathogen free animal. The term “chemically defined” as used herein shall mean, that the medium does not comprise any undefined supplements, such as, for example, extracts of animal components, organs, glands, plants, or yeast. Accordingly, each component of a chemically defined medium is accurately defined. In a preferred embodiment, the media are animal-component free and protein free.
Accordingly, in one aspect, the present invention provides a storage stable, aqueous composition comprising: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 6.0 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 6.0 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 6.0 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a coagulation factor; from 125 mM to 175 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 6.0 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a coagulation factor; from 125 mM to 175 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 6.0 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a coagulation factor; from 125 mM to 175 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 6.0 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a coagulation factor; a stabilizing agent; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; a stabilizing agent; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; a stabilizing agent; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In one embodiment, the present invention provides a storage stable, aqueous composition comprising: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a coagulation factor; an amino acid; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a coagulation factor; an amino acid; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a coagulation factor; an amino acid; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In another aspect, the present invention provides a storage stable, aqueous composition comprising: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM glycine; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM glycine; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM glycine; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a coagulation factor; from 5 mM to 500 mM glycine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a coagulation factor; from 5 mM to 500 mM glycine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a coagulation factor; from 5 mM to 500 mM glycine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In another aspect, the present invention provides a storage stable, aqueous composition comprising: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM proline; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM proline; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM proline; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a coagulation factor; from 5 mM to 500 mM proline; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a coagulation factor; from 5 mM to 500 mM proline; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a coagulation factor; from 5 mM to 500 mM proline; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In another aspect, the present invention provides a storage stable, aqueous composition comprising: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM histidine; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM histidine; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM histidine; and a′ pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a coagulation factor; from 5 mM to 500 mM histidine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a coagulation factor; from 5 mM to 500 mM histidine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a coagulation factor; from 5 mM to 500 mM histidine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In one aspect, the present invention provides a storage stable, aqueous composition comprising: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; an antioxidant; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; an antioxidant; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; an antioxidant; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a coagulation factor; an amino acid; an antioxidant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a coagulation factor; an amino acid; an antioxidant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a coagulation factor; an amino acid; an antioxidant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In one aspect, the present invention provides a storage stable, aqueous composition comprising: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; a sugar and/or sugar alcohol; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; a sugar and/or sugar alcohol; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a coagulation factor; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; a sugar and/or sugar alcohol; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: a coagulation factor; an amino acid; a sugar and/or sugar alcohol; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: a coagulation factor; an amino acid; a sugar and/or sugar alcohol; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: a coagulation factor; an amino acid; a sugar and/or sugar alcohol; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the coagulation factor is a core coagulation factor.
1. Factor VIII
As demonstrated in Example 5, recombinant Factor VIII (rFVIII) is stabilized by the addition of between about 100 mM and about 200 mM of an alkali metal chloride salt (e.g., sodium chloride) at a pH between about 6.0 and about 7.5. In one specific embodiment, methods are provided for the stabilization of Factor VIII formulated at a pH between 6.0 and 7.5, preferably at a pH from 6.5 to 7.0.
In one embodiment, the storage stable, aqueous FVIII compositions provided herein have a protein concentration of between 0.05 g/L and 10 g/L. In certain embodiments, the protein concentration of the FVIII composition is between 0.05 g/L and 2 g/L, or between 0.1 g/L and 1 g/L, or between 0.1 g/L and 0.5 g/L, or any suitable concentration within these ranges, for example 0.05 g/L, or 0.06 g/L, 0.07 g/L, 0.08 g/L, 0.09 g/L, 0.1 g/L, 0.15 g/L, 0.2 g/L, 0.25 g/L, 0.3 g/L, 0.35 g/L, 0.4 g/L, 0.45 g/L, 0.5 g/L, 0.55 g/L, 0.6 g/L, 0.65 g/L, 0.7 g/L, 0.75 g/L, 0.8 g/L, 0.85 g/L, 0.9 g/L, 0.95 g/L, 1 g/L, 1.25 g/L, 1.5 g/L, 1.75 g/L, 2.0 g/L, 2.25 g/L, 2.5 g/L, 2.75 g/L, 3.0 g/L, 3.25 g/L, 3.5 g/L, 3.75 g/L, 4.0 g/L, 4.25 g/L, 4.5 g/L, 4.75 g/L, 5.0 g/L, 5.5 g/L, 6.0 g/L, 6.5 g/L, 7.0 g/L, 7.5 g/L, 8.0 g/L, 8.5 g/L, 9.0 g/L, 9.5 g/L, 10.0 g/L, or higher. In a preferred embodiment, the aqueous FVIII composition will have a concentration of between 0.1 g/L and 0.5 g/L. In yet other embodiments, the concentration of Factor VIII in a storage-stable aqueous formulation is 0.05±0.01 g/L, or 0.06±0.01 g/L, 0.07±0.01 g/L, 0.08±0.01 g/L, 0.09±0.01 g/L, 0.1±0.01 g/L, 0.15±0.01 g/L, 0.2±0.02 g/L, 0.25±0.02 g/L, 0.3±0.03 g/L, 0.35±0.03 g/L, 0.4±0.04 g/L, 0.45±0.04 g/L, 0.5±0.05 g/L, 0.55±0.05 g/L, 0.6±0.06 g/L, 0.65±0.06 g/L, 0.7±0.07 g/L, 0.75±0.07 g/L, 0.8±0.08 g/L, 0.85±0.08 g/L, 0.9±0.09 g/L, 0.95±0.09 g/L, 1±0.1 g/L, 1.25±0.12 g/L, 1.5±0.15 g/L, 1.75±0.17 g/L, 2.0±0.2 g/L, 2.25±0.22 g/L, 2.5±0.25 g/L, 2.75±0.27 g/L, 3.0±0.3 g/L, 3.25±0.32 g/L, 3.5±0.35 g/L, 3.75±0.37 g/L, 4.0±0.4 g/L, 4.25±0.42 g/L, 4.5±0.45 g/L, 4.75±0.47 g/L, 5.0±0.5 g/L, 5.5±0.55 g/L, 6.0±0.6 g/L, 6.5±0.65 g/L, 7.0±0.7 g/L, 7.5±0.75 g/L, 8.0±0.8 g/L, 8.5±0.85 g/L, 9.0±0.9 g/L, 9.5±0.95 g/L, 10.0±1 g/L, or higher.
In one embodiment, the storage stable, aqueous FVIII composition will be stable under refrigeration (i.e., between about 2° C. and about 8° C.) for at least about 1 month. In other embodiments, the storage stable, aqueous FVIII composition will be stable under refrigeration for at least about 2 months. In a preferred embodiment, the storage stable, aqueous FVIII composition will be stable under refrigeration for at least about 3 months. In yet another embodiment, the storage stable, aqueous FVIII composition will be stable under refrigeration for at least about 6 months. In yet other embodiments, the storage stable, aqueous FVIII composition will be stable under refrigeration for at least about 2 weeks, or at least about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 weeks or at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or more months under refrigeration.
Accordingly, in one aspect, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 6.0 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 6.0 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 6.0 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; from 125 mM to 175 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 6.0 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; from 125 mM to 175 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 6.0 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; from 125 mM to 175 mM of an alkali metal chloride salt; a stabilizing agent; and a pH of from 6.0 to 7.5. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; a stabilizing agent; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting essentially of: a labile therapeutic protein; a stabilizing agent; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein consisting of: a labile therapeutic protein; a stabilizing agent; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a more specific embodiment, the stabilizing agent is an amino acid. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In one embodiment, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; an amino acid; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; an amino acid; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; an amino acid; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In another aspect, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM glycine; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM glycine; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM glycine; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; from 5 mM to 500 mM glycine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; from 5 mM to 500 mM glycine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; from 5 mM to 500 mM glycine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM glycine. In a more specific embodiment, the composition contains from 100 mM to 400 mM glycine. In a more specific embodiment, the composition contains from 150 mM to 350 mM glycine. In a more specific embodiment, the composition contains from 200 mM to 300 mM glycine. In a more specific embodiment, the composition contains 250±25 mM glycine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In another aspect, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM proline; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM proline; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM proline; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; from 5 mM to 500 mM proline; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; from 5 mM to 500 mM proline; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; from 5 mM to 500 mM proline; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM proline. In a more specific embodiment, the composition contains from 100 mM to 400 mM proline. In a more specific embodiment, the composition contains from 150 mM to 350 mM proline. In a more specific embodiment, the composition contains from 200 mM to 300 mM proline. In a more specific embodiment, the composition contains 250±25 mM proline. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In another aspect, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM histidine; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM histidine; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; from 5 mM to 500 mM histidine; and a pH of from 6.0 to 7.5. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; from 5 mM to 500 mM histidine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; from 5 mM to 500 mM histidine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; from 5 mM to 500 mM histidine; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the composition contains from 25 mM to 500 mM histidine. In a more specific embodiment, the composition contains from 100 mM to 400 mM histidine. In a more specific embodiment, the composition contains from 150 mM to 350 mM histidine. In a more specific embodiment, the composition contains from 200 mM to 300 mM histidine. In a more specific embodiment, the composition contains 250±25 mM histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In one aspect, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; an antioxidant; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; an antioxidant; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; an antioxidant; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; an amino acid; an antioxidant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; an amino acid; an antioxidant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; an amino acid; an antioxidant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In one aspect, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; a sugar and/or sugar alcohol; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; a sugar and/or sugar alcohol; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; a sugar and/or sugar alcohol; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In another embodiment, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; an amino acid; a sugar and/or sugar alcohol; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; an amino acid; a sugar and/or sugar alcohol; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; an amino acid; a sugar and/or sugar alcohol; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains calcium.
In one aspect, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; mannitol; trehalose; calcium; a non-ionic surfactant; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains an antioxidant. In a specific embodiment, the concentration of mannitol is 38±7 g/L. In another specific embodiment, the concentration of trehalose is 10±2 g/L. In another specific embodiment, the concentration of histidine is 12±2 mM. In another specific embodiment, the concentration of calcium is 1.9±0.4 mM. In another embodiment, the non-ionic surfactant is polysorbate-80 at a concentration of 0.15±0.03 g/L.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; mannitol; trehalose; calcium; a non-ionic surfactant; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains an antioxidant. In a specific embodiment, the concentration of mannitol is 38±7 g/L. In another specific embodiment, the concentration of trehalose is 10±2 g/L. In another specific embodiment, the concentration of histidine is 12±2 mM. In another specific embodiment, the concentration of calcium is 1.9±0.4 mM. In another embodiment, the non-ionic surfactant is polysorbate-80 at a concentration of 0.15±0.03 g/L.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; from 75 mM to 200 mM of an alkali metal chloride salt; an amino acid; mannitol; trehalose; calcium; a non-ionic surfactant; Tris; glutathione; and a pH of from 6.0 to 7.5. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific, embodiment, the composition also contains an antioxidant. In a specific embodiment, the concentration of mannitol is 38±7 g/L. In another specific embodiment, the concentration of trehalose is 10±2 g/L. In another specific embodiment, the concentration of histidine is 12±2 mM. In another specific embodiment, the concentration of calcium is 1.9±0.4 mM. In another embodiment, the non-ionic surfactant is polysorbate-80 at a concentration of 0.15±0.03 g/L.
In one aspect, the present invention provides a storage stable, aqueous composition comprising: Factor VIII; an amino acid; mannitol; trehalose; calcium; a non-ionic surfactant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains an antioxidant. In a specific embodiment, the concentration of mannitol is 38±7 g/L. In another specific embodiment, the concentration of trehalose is 10±2 g/L. In another specific embodiment, the concentration of histidine is 12±2 mM. In another specific embodiment, the concentration of calcium is 1.9±0.4 mM. In another embodiment, the non-ionic surfactant is polysorbate-80 at a concentration of 0.15±0.03 g/L.
In a specific embodiment, the present invention provides a storage stable, aqueous composition consisting essentially of: Factor VIII; an amino acid; mannitol; trehalose; calcium; a non-ionic surfactant; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains an antioxidant. In a specific embodiment, the concentration of mannitol is 38±7 g/L. In another specific embodiment, the concentration of trehalose is 10±2 g/L. In another specific embodiment, the concentration of histidine is 12±2 mM. In another specific embodiment, the concentration of calcium is 1.9±0.4 mM. In another embodiment, the non-ionic, surfactant is polysorbate-80 at a concentration of 0.15±0.03 g/L.
In a more specific embodiment, the present invention provides a storage stable, aqueous composition consisting of: Factor VIII; an amino acid; mannitol; trehalose; calcium; a non-ionic surfactant; Tris; glutathione; and an alkali metal salt/pH combination selected from any one of variations 1 to 2088, as set forth in Table 1, Table 2, Table 3, and Table 4. In a specific embodiment, the amino acid is glycine, proline, or histidine. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline. In another preferred embodiment, the amino acid is histidine. In one embodiment, the metal chloride salt is sodium chloride. In another embodiment, the metal chloride salt is potassium chloride. In a preferred embodiment, the pH of the formulation is between 6.5 and 7.5. In a specific embodiment, the composition also contains an antioxidant. In a specific embodiment, the concentration of mannitol is 38±7 g/L. In another specific embodiment, the concentration of trehalose is 10±2 g/L. In another specific embodiment, the concentration of histidine is 12±2 mM. In another specific embodiment, the concentration of calcium is 1.9±0.4 mM. In another embodiment, the non-ionic surfactant is polysorbate-80 at a concentration of 0.15±0.03 g/L.
In the context of the present invention, a labile therapeutic protein is unstable when formulated at mildly acidic to neutral pH in the absence of an alkaline metal chloride salt. Surprisingly, it has been found that a wide range of labile therapeutic proteins are stabilized by the addition of a moderate concentration (i.e., between about 75 mM and about 200 mM, preferably between about 100 mM and about 200 mM) of an alkaline metal chloride salt. This effect of alkali metal chloride salts provides methods for stabilizing aqueous formulations of labile therapeutic proteins. Accordingly, in one aspect of the present invention, methods are provided for the stabilization of an aqueous labile therapeutic protein composition. These methods allow for aqueous formulations of labile therapeutic proteins at mildly acidic to neutral pH, which previously required lyophilization, freezing in the presence of several stabilizers, or formulation at extreme pH values.
In one embodiment, the method comprises the addition of alkali metal chloride salt to a final concentration of between about 75 mM and about 200 mM to an aqueous formulation of a labile therapeutic protein at a pH between about 5.5 and about 7.5. In a preferred embodiment, the method further comprises the addition of a stabilizing agent, such as an amino acid, to the formulation.
In another embodiment, the method comprises the addition of alkali metal chloride salt to a final concentration of between about 100 mM and about 200 mM to an aqueous formulation of a labile therapeutic protein at a pH between about 5.5 and about 7.5. In a preferred embodiment, the method further comprises the addition of a stabilizing agent, such as an amino acid, to the formulation.
In another embodiment, the method comprises the addition of alkali metal chloride salt to a final concentration of between about 100 mM and about 200 mM to an aqueous formulation of a labile therapeutic protein at a pH between about 5.5 and about 7.0. In a preferred embodiment, the method further comprises the addition of a stabilizing agent, such as an amino acid, to the formulation.
In yet another embodiment, the method comprises the addition of alkali metal chloride salt to a final concentration of between about 75 mM and about 200 mM to an aqueous formulation of a labile therapeutic protein at a pH between about 5.5 and about 7.0. In a preferred embodiment, the method further comprises the addition of a stabilizing agent, such as an amino acid, to the formulation.
In certain embodiments, the methods provided herein for the stabilization of a labile therapeutic protein will further comprise the addition of a stabilizing agent to the formulation. In a preferred embodiment, the stabilizing agent will be an amino acid. Exemplary amino acids that may be used for this purpose include, without limitation, arginine, histidine, lysine, serine, proline, glycine, alanine, threonine, and a combination thereof. In a preferred embodiment, the amino acid is glycine. In another preferred embodiment, the amino acid is proline.
For purposes of further stabilizing the compositions provided herein, the amino acid will typically be added to the formulation at a concentration between about 25 mM and about 0.75 M. In one embodiment, at least about 100 mM of the amino acid is added to the formulation. In another embodiment, at least about 200 mM of the amino acid is added to the formulation. In yet another embodiment, at least about 250 mM of the amino acid is added to the formulation. In yet other embodiments, the formulations provided herein will contain at least about 25 mM of the amino acid, or at least about 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, or more of the amino acid.
In a preferred embodiment, the labile therapeutic protein is a human protein, a humanized protein, or a chimeric human protein. The human protein may be either purified from a natural source (e.g., pooled human plasma) or expressed recombinantly, for example in a mammalian cell or tissue culture. In a preferred embodiment, the labile therapeutic protein is a plasma-derived protein, preferably a plasma-derived coagulation factor or immunoglobulin preparation. In another preferred embodiment, the human, humanized, or chimeric protein in a recombinant antibody or fragment thereof.
In one embodiment, the methods for stabilizing a formulation of a labile therapeutic protein provided herein comprise the addition of between about 75 mM and about 200 mM of an alkali metal chloride salt to the formulation. In a preferred embodiment, the methods for stabilizing a formulation of a labile therapeutic protein provided herein comprise the addition of between about 100 mM and about 200 mM of an alkali metal chloride salt to the formulation. In certain embodiments, the methods comprise the addition of between about 150 mM and about 200 mM of an alkali metal chloride salt. In other embodiments, the method comprises the addition of between about 150 mM and about 200 mM of an alkali metal chloride salt. In yet other embodiments, the comprise the addition of at or about 70 mM or at or about 75 mM, 80 mM, 85 mM, 90 mM, 95 mM, 100 mM, 105 mM, 110 mM, 115 mM, 120 mM, 125 mM, 130 mM, 135 mM, 140 mM, 145 mM, 150 mM, 155 mM, 160 mM, 165 mM, 170 mM, 175 mM, 180 mM, 185 mM, 190 mM, 195 mM, 200 mM, 205 mM, 210 mM, 215 mM, or at or about 220 mM of an alkali metal chloride salt. In one preferred embodiment, the alkali metal chloride salt is sodium chloride. In another preferred embodiment, the alkali metal chloride salt is potassium chloride.
The methods for stabilizing a labile therapeutic protein provided by the present invention comprise formulating the labile protein, in the presence of an alkali metal chloride salt, at a mildly acidic to neutral pH. Generally, this includes pH values between about 5.5 and about 7.5. However, the range of pH values at which any individual labile therapeutic protein is stabilized by the addition of a moderate level (i.e., between about 75 mM and about 200 mM, preferably between about 100 mM and about 200 mM) of an alkali metal chloride salt may vary slightly, dependent upon the properties of the individual protein. In a preferred embodiment, the method will comprise formulating a storage stable composition of a labile therapeutic protein at a pH between about 5.5 and about 7.0. In another embodiment, the method will comprise formulating a storage stable composition of a labile therapeutic protein at a pH between about 5.5 and about 6.5. In other embodiments, the pH of the stabilizing formulation will be between about 6.0 and about 7.0. In another embodiment, the pH of the stabilizing formulation will be between about 5.5 and about 6.0. In one embodiment, the pH of the stabilizing formulation will be between about 6.0 and about 6.5. In another embodiment, the pH of the stabilizing formulation will be between about 6.5 and about 7.0. In another embodiment, the pH of the stabilizing formulation will be between about 6.0 and about 7.5. In another embodiment, the pH of the stabilizing formulation will be between about 6.5 and about 7.5. In another embodiment, the pH of the stabilizing formulation will be between about 7.0 and about 7.5. In yet other embodiments, the pH of the stabilizing formulation may be at or about 5.5, or at or about 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, or at or about 7.5.
In certain embodiments, the method will further comprise formulating the storage stable, labile therapeutic protein for parenteral administration including, but not limited to, intradermal, subcutaneous, transdermal implant, intracavernous, intravitreal, transscleral, intracerebral, intrathecal, epidural, intravenous, intracardiac, intramuscular, intraosseous, intraperitoneal, and nanocell injection administration. In one preferred embodiment, the compositions provided herein will be formulated for intravenous administration. In another preferred embodiment, the compositions provided herein will be formulated for subcutaneous administration. In yet another preferred embodiment, the compositions provided herein will be formulated for intramuscular administration.
In certain embodiments, the method for stabilizing a labile therapeutic protein will comprise formulating the protein at a final concentration of between about 0.05 mg/mL to about 250 mg/mL. In certain embodiments, the labile protein will be formulated at a final concentration of about 0.05 mg/mL, 0.06 mg/mL, 0.07 mg/mL, 0.08 mg/mL, 0.09 mg/mL, 0.1 mg/mL, 0.2 mg/mL, 0.3 mg/mL, 0.4 mg/mL, 0.5 mg/mL or about 1 mg/mL, 2 mg/mL, 3 mg/mL, 4 mg/mL, 5 mg/mL, 6 mg/mL, 7 mg/mL, 8 mg/mL, 9 mg/mL, 10 mg/mL, 11 mg/mL, 12 mg/mL, 13 mg/mL, 14 mg/mL, 15 mg/mL, 16 mg/mL, 17 mg/mL, 18 mg/mL, 19 mg/mL, 20 mg/mL, 21 mg/mL, 22 mg/mL, 23 mg/mL, 24 mg/mL, 25 mg/mL, 26 mg/mL, 27 mg/mL, 28 mg/mL, 29 mg/mL, 30 mg/mL, 35 mg/mL, 40 mg/mL, 45 mg/mL, 50 mg/mL, 55 mg/mL, 60 mg/mL, 65 mg/mL, 70 mg/mL, 75 mg/mL, 80 mg/mL, 85 mg/mL, 90 mg/mL, 95 mg/mL, 100 mg/mL, 110 mg/mL, 120 mg/mL, 130 mg/mL, 140 mg/mL, 150 mg/mL, 160 mg/mL, 170 mg/mL, 180 mg/mL, 190 mg/mL, 200 mg/mL, 210 mg/mL, 220 mg/mL, 230 mg/mL, 240 mg/mL, 250 mg/mL, or higher concentrations, depending upon the characteristics of the protein being formulated, the intended therapeutic use of the protein, and the preferred method of administration.
In one embodiment, the method will comprise formulating the labile therapeutic protein at a low final protein concentration of between about 0.05 mg/mL and about 20 mg/mL. In another embodiment, the final protein concentration may be between about 0.5 mg/mL and about 15 mg/mL. In another embodiment, the final protein concentration may be between about 0.5 mg/mL and about 10 mg/mL. In another embodiment, the final protein concentration may be between about 0.5 mg/mL and about 5 mg/mL. In one embodiment, a composition with a final protein concentration as described above will be formulated for intravenous administration.
In other embodiments, the method will comprise formulating the labile therapeutic protein at a moderate final protein concentration of between about 5 mg/mL and about 25 mg/mL. In another embodiment, the final protein concentration may be between about 10 mg/mL and about 25 mg/mL. In another embodiment, the final protein concentration may be between about 15 mg/mL and about 25 mg/mL. In another embodiment, the final protein concentration may be between about 20 mg/mL and about 25 mg/mL. In one embodiment, a composition with a final protein concentration as described above will be formulated for subcutaneous or intramuscular administration.
In certain embodiments, the final protein concentration may be between about 0.5% and about 25%. In another embodiment, the final protein concentration may be between about 0.5% and about 20%. In another embodiment, the final protein concentration may be between about 0.5% and about 15%. In another embodiment, the final protein concentration may be between about 0.5% and about 10%. In another embodiment, the final protein concentration may be between about 0.5% and about 5%. In one embodiment, a composition with a final protein concentration as described above will be formulated for intravenous administration.
In certain embodiments, the final protein concentration may be between about 5% and about 25%. In another embodiment, the final protein concentration may be between about 10% and about 25%. In another embodiment, the final protein concentration may be between about 15% and about 25%. In another embodiment, the final protein concentration may be between about 20% and about 25%. In one embodiment, a composition with a final protein concentration as described above will be formulated for subcutaneous or intramuscular administration.
In one embodiment, the present invention provides a method for stabilizing an aqueous formulation of a labile plasma derived protein at a pH between about 5.5 and about 7.5, the method comprising adding an alkali metal chloride salt to the formulation at a final concentration of between about 75 mM and about 200 mM. In a preferred embodiment, the compositions comprise between about 100 mM and about 200 mM of an alkali metal chloride salt. In another preferred embodiment, the mildly acidic to neutral pH is between about 5.5 and about 7.0. In a preferred embodiment, the alkali metal chloride salt is sodium chloride. In another preferred embodiment, the alkali metal chloride salt is potassium chloride.
The methods provided herein allow for the stabilization of a labile protein at a mildly acidic to neutral pH for an extended period of time. For example, in one embodiment, the storage stable, aqueous labile therapeutic protein composition will be stable for at least about 2 months. In another embodiment, the composition will be stable for at least about 3 months. In yet other embodiment, the composition will be stable for at least 1 about month, or at least about 2, 3, 4, 5, 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, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or more months. In a preferred embodiment, the composition will be stable for at least about 6 months. In a more preferred embodiment, the composition will be stable for at least about 1 year. In a more preferred embodiment, the composition will be stable for at least about 2 years.
Dependent upon the individual characteristics of the labile protein being stabilized in the methods provided herein, the formulation will be stabile for an extended period of time at a temperature between about 2° C. and about 42° C. In one embodiments, a labile therapeutic protein will be stabilized by the methods provided herein when stored under refrigeration, i.e., stored at a temperature between about 2° C. and about 8° C. In another embodiment, a labile therapeutic protein will be stabilized by the methods provided herein when stored at room temperature, i.e., stored at a temperature between about 20° C. and about 25° C. In other embodiments, the protein may be stabilized when stored at a temperature between about 28° C. and about 32° C. In yet another embodiment, the protein may be stabilized when stored at a temperature between about 38° C. and about 42° C. The temperatures at which a labile therapeutic protein will be stabilized by the methods provided herein will be dependent upon the characteristics of the individual protein, which can readily be determined by one of skill in the art.
In certain embodiments, the methods provided herein for the stabilization of a labile therapeutic protein comprise the addition of a stabilizing agent. In one embodiment, the stabilizing agent comprises one or more buffering agents or pH stabilizing agents suitable for intravenous, intravitreal, subcutaneous, and/or intramuscular administration. Non-limiting examples of buffering agents suitable for formulating the storage stable compositions provided herein include glycine, histidine, or other amino acids, salts like citrate, phosphate, acetate, glutamate, tartrate, benzoate, lactate, gluconate, malate, succinate, formate, propionate, carbonate, or any combination thereof adjusted to an appropriate pH. Generally, the buffering agent will be sufficient to maintain a suitable pH in the formulation for an extended period of time.
In some embodiments, the concentration of buffering agent in the formulation will be at or about between 5 mM and 500 mM. In certain embodiments, the concentration of the buffering agent in the formulation will be at or about 5 mM, 10 mM, 15 mM, 20 mM, 25 mM, 50 mM, 75 mM, 100 mM, 125 mM, 150 mM, 175 mM, 200 mM, 225 mM, 250 mM, 275 mM, 300 mM, 325 mM, 350 mM, 375 mM, 400 mM, 425 mM, 450 mM, 475 mM, 500 mM or higher.
In another embodiment, the stabilizing agent will comprise an agent for adjusting the osmolarity of the composition. Non-limiting examples of osmolarity agents include mannitol, sorbitol, glycerol, sucrose, glucose, dextrose, levulose, fructose, lactose, polyethylene glycols, phosphates, calcium chloride, calcium gluconoglucoheptonate, dimethyl sulfone, and the like.
In a preferred embodiment, the stabilizing agent employed in the storage stable, labile immunoglobulin formulations provided herein will be an amino acid. Stabilizing amino acids include arginine, histidine, lysine, serine, proline, glycine, alanine, threonine, and a combination thereof. In a preferred embodiment, the amino acid is glycine. For purposes of further stabilizing the compositions provided herein, the amino acid will typically be added to the formulation at a concentration between about 25 mM and about 0.75 M. In one embodiment, at least about 100 mM of the amino acid is added to the formulation. In another embodiment, at least about 200 mM of the amino acid is added to the formulation. In yet another embodiment, at least about 250 mM of the amino acid is added to the formulation. In yet other embodiments, the formulations provided herein will contain at least about 25 mM of the amino acid, or at least about 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, or more of the amino acid.
In a specific embodiment, the labile plasma derived protein is an immunoglobulin preparation. As demonstrated in Examples 1 and 2, plasma derived IgG preparations are stabilized by the addition of between about 100 mM and about 200 mM of an alkali metal chloride salt (e.g., sodium chloride) at a pH between about 6.0 and about 7.0. As shown in
Accordingly, in one embodiment, the present invention provides method for stabilizing a plasma derived IgG aqueous composition formulated at a pH between about 6.0 and about 7.0, the method comprising adding between about 75 mM and about 200 mM, preferably between about 100 mM and about 200 mM of an alkali metal chloride salt to the formulation. In a preferred embodiment, the method further comprises adding a stabilizing agent to the formulation. In another preferred embodiment, the method comprises the addition of between about 75 mM and about 200 mM, preferably between about 100 mM and about 200 mM, of an alkali metal chloride salt to a plasma derived immunoglobulin preparation formulated at a pH between about 6.0 and about 6.5. In a preferred embodiment, the salt is sodium chloride. In another preferred embodiment, the salt is potassium chloride.
In one embodiment, the method for stabilizing a labile plasma derived immunoglobulin composition will comprise formulating the protein at a final concentration of between about 30 g/L and about 250 g/L. In certain embodiments, the method comprises formulating the immunoglobulin composition at a final protein concentration of between about 50 g/L and about 200 g/L, or between about 70 g/L and about 150 g/L, or between about 90 g/L and about 120 g/L, or any suitable concentration within these ranges, for example about 30 g/L, or about 35 g/L, 40 g/L, 45 g/L, 50 g/L, 55 g/L, 60 g/L, 65 g/L, 70 g/L, 75 g/L, 80 g/L, 85 g/L, 90 g/L, 95 g/L, 100 g/L, 105 g/L, 110 g/L, 115 g/L, 120 g/L, 125 g/L, 130 g/L, 135 g/L, 140 g/L, 145 g/L, 150 g/L, 155 g/L, 160 g/L, 165 g/L, 170 g/L, 175 g/L, 180 g/L, 185 g/L, 190 g/L, 195 g/L, 200 g/L, 205 g/L, 210 g/L, 215 g/L, 220 g/L, 225 g/L, 230 g/L, 235 g/L, 240 g/L, 245 g/L, 250 g/L, or higher. In a preferred embodiment, the aqueous IgG composition will have a concentration of at or about 100 g/L. In a related embodiment, the aqueous IgG composition will have a concentration of between about 70 g/L and about 130 g/L. In another preferred embodiment, the aqueous IgG composition will have a concentration of at or about 200 g/L. In a related embodiment, the aqueous IgG composition will have a concentration of between about 170 g/L and about 230 g/L.
In one aspect, the present invention provides methods for stabilizing recombinant immunoglobulin preparations at a pH between about 5.5 and about 7.5 comprising the addition of between about 75 mM and about 200 mM of an alkali metal chloride salt to the formulation. In a preferred embodiment, the compositions comprise between about 100 mM and about 200 mM of an alkali metal chloride salt. In another preferred embodiment, the mildly acidic to neutral pH is between about 5.5 and about 7.0. In a preferred embodiment, the methods further comprise the addition of a stabilizing agent to the formulation. In one preferred embodiment, the salt is sodium chloride. In another preferred embodiment, the salt is potassium chloride.
In another specific embodiment, the storage stable, immunoglobulin composition is a recombinant antibody preparation. As demonstrated in Example 4, a recombinant anti-MIF monoclonal antibody preparation is stabilized by the addition of between about 100 mM and about 200 mM of an alkali metal chloride salt (e.g., sodium chloride) at a pH between about 5.5 and about 6.5.
Accordingly, in one embodiment, the present invention provides methods for stabilizing aqueous compositions of recombinant immunoglobulins formulated at a pH between about 5.5 and about 6.5 by the addition of between about 75 mM and about 200 mM, preferably between about 100 mM and about 200 mM, of an alkali metal chloride salt and optionally, a stabilizing agent to the formulation. In a preferred embodiment, the salt is sodium chloride. In another preferred embodiment, the salt is potassium chloride.
In one embodiment, the methods for stabilizing a recombinant immunoglobulin aqueous composition provided herein comprise the formulation of the recombinant immunoglobulin at a final protein concentration of between about 1 g/L and about 250 g/L. In certain embodiments, the protein concentration of the recombinant immunoglobulin composition is between about 50 g/L and about 200 g/L, or between about 70 g/L and about 150 g/L, or between about 90 g/L and about 120 g/L, or any suitable concentration within these ranges, for example about 1 g/L, 2 g/L, 3 g/L, 4 g/L, 5 g/L, 6 g/L, 7 g/L, 8 g/L, 9 g/L, 10 g/L, 11 g/L, 12 g/L, 13 g/L, 14 g/L, 15 g/L, 16 g/L, 17 g/L, 18 g/L, 19 g/L, 20 g/L, 25 g/L, 30 g/L, or about 35 g/L, 40 g/L, 45 g/L, 50 g/L, 55 g/L, 60 g/L, 65 g/L, 70 g/L, 75 g/L, 80 g/L, 85 g/L, 90 g/L, 95 g/L, 100 g/L, 105 g/L, 110 g/L, 115 g/L, 120 g/L, 125 g/L, 130 g/L, 135 g/L, 140 g/L, 145 g/L, 150 g/L, 155 g/L, 160 g/L, 165 g/L, 170 g/L, 175 g/L, 180 g/L, 185 g/L, 190 g/L, 195 g/L, 200 g/L, 205 g/L, 210 g/L, 215 g/L, 220 g/L, 225 g/L, 230 g/L, 235 g/L, 240 g/L, 245 g/L, 250 g/L, or higher.
In another embodiment, the present invention provides methods for stabilizing a labile coagulation factor formulated at a pH between about 5.5 and about 7.5 comprising the addition of between about 75 mM and about 200 mM of an alkali metal chloride salt to the formulation. In a preferred embodiment, the compositions comprise between about 100 mM and about 200 mM of an alkali metal chloride salt. In another preferred embodiment, the mildly acidic to neutral pH is between about 5.5 and about 7.0. In a preferred embodiment, the methods further comprise the addition of a stabilizing agent to the formulation. In one preferred embodiment, the salt is sodium chloride. In another preferred embodiment, the salt is potassium chloride.
In certain embodiments, the labile coagulation protein is a plasma derived protein or preparation. In other embodiments, the labile coagulation protein is a recombinantly expressed coagulation protein. Methods for manufacturing recombinant and plasma derived coagulation factors are well known in the art. Non-limiting examples of coagulation factors that may be formulated according to the methods provided herein include, Factor II (prothrombin), Factor III (platelet tissue factor), Factor V, Factor VII, Factor VIII, Factor IX, Factor X, Factor XI, Factor XII, Factor XIII, von Willebrand Factor (vWF), and the like. In a preferred embodiment, the labile coagulation protein is selected from Factor VII, Factor VIII, Factor IX, and von Willebrand Factor (vWF). In another preferred embodiment, the labile coagulation protein is a vitamin-K dependent protein complex, for example, comprising Factor II, Factor IX, and Factor X, or comprising Factor II, Factor VII, Factor IX, and Factor X.
In one specific embodiment, methods are provided for the stabilization of Factor VIII formulated at a pH between about 6.5 and about 7.0. As shown in Example 5, recombinant Factor VIII (rFVIII) is stabilized by the addition of between about 100 mM and about 200 mM of an alkali metal chloride salt (e.g., sodium chloride) at a pH between about 6.5 and about 7.0.
Accordingly, in one embodiment the present invention provides a method for stabilizing an aqueous composition of FVIII, the method comprising formulating a FVIII composition at a pH between about 6.5 and about 7.0 with between about 75 mM and about 200 mM, preferably between about 100 mM and about 200 mM, of an alkali metal chloride salt. In a preferred embodiment, the alkali metal chloride salt is sodium chloride. In another preferred embodiment, the alkali metal chloride salt is potassium chloride.
In one embodiment, the present invention provides a method for stabilizing an aqueous formulation of a labile therapeutic protein at a pH between about 5.5 and about 7.5, the method comprising addition of an alkali metal chloride salt at a concentration of between about 75 mM and about 200 mM. In a preferred embodiment, the compositions comprise between about 100 mM and about 200 mM of an alkali metal chloride salt. In another preferred embodiment, the mildly acidic to neutral pH is between about 5.5 and about 7.0. In a preferred embodiment, the salt is sodium chloride. In certain embodiments, the method further comprises the addition of a stabilizing agent. In one embodiment, the stabilizing agent is an amino acid. In preferred embodiments, the amino acid is glycine or proline.
In another embodiment, the present invention provides a method for stabilizing an aqueous formulation of a labile therapeutic protein at a pH between about 5.5 and about 6.5, the method comprising addition of an alkali metal chloride salt at a concentration of between about 100 mM and about 200 mM. In a preferred embodiment, the salt is sodium chloride. In certain embodiments, the method further comprises the addition of a stabilizing agent. In one embodiment, the stabilizing agent is an amino acid. In preferred embodiments, the amino acid is glycine or proline.
In one embodiment of the methods provided herein, the labile therapeutic protein is a human or humanized protein. In another embodiment of the methods provided herein, the labile therapeutic protein is a recombinant protein. In yet another embodiment of the methods provided herein, the labile therapeutic protein is a plasma-derived protein.
In a specific embodiment of the methods provided herein, the labile therapeutic protein is an immunoglobulin. In one embodiment, the immunoglobulin is an IgG preparation. In another embodiment, the immunoglobulin is a recombinant antibody.
In one embodiment, the present invention provides a method for stabilizing an aqueous formulation of a labile immunoglobulin preparation at a pH between about 5.5 and about 7.5, the method comprising addition of an alkali metal chloride salt at a concentration of between about 75 mM and about 200 mM, wherein the method stabilizes the immunoglobulin composition for at least 6 months when stored at a temperature at or below about 42° C., for example, at a temperature between about 38° C. and about 42° C. In a preferred embodiment, the pH of the immunoglobulin formulation is between about 5.5 and about 6.5.
In one embodiment, the present invention provides a method for stabilizing an aqueous formulation of a labile immunoglobulin preparation at a pH between about 5.5 and about 7.0, the method comprising addition of an alkali metal chloride salt at a concentration of between about 100 mM and about 200 mM, wherein the method stabilizes the immunoglobulin composition for at least 6 months when stored at a temperature at or below about 42° C., for example, at a temperature between about 38° C. and about 42° C. In a preferred embodiment, the pH of the immunoglobulin formulation is between about 5.5 and about 6.5.
In one embodiment, the present invention provides a method for stabilizing an aqueous formulation of a labile immunoglobulin preparation at a pH between about 5.5 and about 7.0, the method comprising addition of an alkali metal chloride salt at a concentration of between about 100 mM and about 200 mM, wherein the method stabilizes the immunoglobulin composition for at least about one year when stored at a temperature at or below about 32° C., for example, at a temperature between about 28° C. and about 32° C. In a preferred embodiment, the pH of the immunoglobulin formulation is between about 5.5 and about 6.5.
In one embodiment, the present invention provides a method for stabilizing an aqueous formulation of a labile immunoglobulin preparation at a pH between about 5.5 and about 7.5, the method comprising addition of an alkali metal chloride salt at a concentration of between about 75 mM and about 200 mM, wherein the method stabilizes the immunoglobulin composition for at least about one year when stored at a temperature at or below about 32° C., for example, at a temperature between about 28° C. and about 32° C. In a preferred embodiment, the pH of the immunoglobulin formulation is between about 5.5 and about 6.5.
In a specific embodiment of the methods provided herein, the labile therapeutic protein is a labile coagulation protein. In one embodiment, the coagulation protein is Factor VIII. In another embodiment, the coagulation protein is Factor VII. In yet another embodiment, the coagulation protein is Factor IX. In another embodiment, the coagulation protein is a protein K-dependent coagulation complex, for example, comprising Factor II, Factor IX, and Factor X, or comprising Factor II, Factor VII, Factor IX, and Factor X.
In one embodiment, the present invention provides a method for stabilizing an aqueous formulation of a labile coagulation protein at a pH between about 5.5 and about 7.5, the method comprising addition of an alkali metal chloride salt at a concentration of between about 75 mM and about 200 mM, wherein the method stabilizes the coagulation protein for at least 3 months when stored at a refrigerated temperature, for example, at a temperature between about 2° C. and about 10° C.
In one embodiment, the present invention provides a method for stabilizing an aqueous formulation of a labile coagulation protein at a pH between about 5.5 and about 7.0, the method comprising addition of an alkali metal chloride salt at a concentration of between about 100 mM and about 200 mM, wherein the method stabilizes the coagulation protein for at least 3 months when stored at a refrigerated temperature, for example, at a temperature between about 2° C. and about 10° C.
In one embodiment, the present invention provides a method for stabilizing an aqueous formulation of a labile coagulation protein at a pH between about 5.5 and about 7.0, the method comprising addition of an alkali metal chloride salt at a concentration of between about 100 mM and about 200 mM, wherein the method stabilizes the coagulation protein for at least 6 months when stored at a refrigerated temperature, for example, at a temperature between about 2° C. and about 10° C.
In one embodiment, the present invention provides a method for stabilizing an aqueous formulation of a labile coagulation protein at a pH between about 5.5 and about 7.5, the method comprising addition of an alkali metal chloride salt at a concentration of between about 75 mM and about 200 mM, wherein the method stabilizes the coagulation protein for at least 6 months when stored at a refrigerated temperature, for example, at a temperature between about 2° C. and about 10° C.
In one embodiment, the present invention provides a method for stabilizing an aqueous formulation of a labile coagulation protein at a pH between about 5.5 and about 7.0, the method comprising addition of an alkali metal chloride salt at a concentration of between about 100 mM and about 200 mM, wherein the method stabilizes the coagulation protein for at least about one year when stored at a refrigerated temperature, for example, at a temperature between about 2° C. and about 10° C.
In one embodiment, the present invention provides a method for stabilizing an aqueous formulation of a labile coagulation protein at a pH between about 5.5 and about 7.5, the method comprising addition of an alkali metal chloride salt at a concentration of between about 75 mM and about 200 mM, wherein the method stabilizes the coagulation protein for at least about one year when stored at a refrigerated temperature, for example, at a temperature between about 2° C. and about 10° C.
In one embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: between about 75 mM and about 200 mM of an alkali metal chloride salt; a stabilizing agent; and a pH between about 5:5 and about 7.5.
In one embodiment, the present invention provides a storage stable, aqueous composition of a labile therapeutic protein comprising: between about 100 mM and about 200 mM of an alkali metal chloride salt; a stabilizing agent; and a pH between about 5.5 and about 7.0.
In a specific embodiment of the compositions provided above, the labile therapeutic protein is a human or humanized protein.
In a specific embodiment of the compositions provided above, the protein is a recombinant protein.
In a specific embodiment of the compositions provided above, the protein is a plasma-derived protein.
In a specific embodiment of the compositions provided above, the protein is an immunoglobulin.
In a specific embodiment of the compositions provided above, the immunoglobulin is an IgG preparation.
In a specific embodiment of the compositions provided above, the protein concentration of the IgG preparation is at least about 100 mg/mg.
In a specific embodiment of the compositions provided above, the protein concentration of the IgG preparation is at least about 200 mg/mL.
In a specific embodiment of the compositions provided above, the immunoglobulin is a recombinant antibody.
In a specific embodiment of the compositions provided above, the composition is stable for at least 6 months when stored at between about 38° C. and about 42° C.
In a specific embodiment of the compositions provided above, the composition is stable for at least 1 year when stored at between about 28° C. and about 32° C.
In a specific embodiment of the compositions provided above, the pH of the composition is between about 5.5 and about 6.5.
In a specific embodiment of the compositions provided above, the protein is a coagulation factor.
In a specific embodiment of the compositions provided above, the coagulation factor is Factor VIII.
In a specific embodiment of the compositions provided above, the pH of the composition is between about 6.0 and about 7.0.
In a specific embodiment of the compositions provided above, the pH of the composition is 6.5±0.2.
In a specific embodiment of the compositions provided above, the composition retains at least 80% of its Factor VIII activity when stored at a temperature between about 2° C. and about 8° C. for at least 3 months.
In a specific embodiment of the compositions provided above, the coagulation factor is Factor VII.
In a specific embodiment of the compositions provided above, the coagulation factor is Factor IX.
In a specific embodiment of the compositions provided above, the coagulation factor is von Willebrand Factor (vWF).
In a specific embodiment of the compositions provided above, the coagulation factor is a protein K-dependent coagulation complex.
In a specific embodiment of the compositions provided above, the protein K-dependent coagulation complex comprises the coagulation factors Factor II, Factor IX, and Factor X.
In a specific embodiment of the compositions provided above, the protein K-dependent coagulation complex further comprises Factor VII.
In a specific embodiment of the compositions provided above, the labile protein is stable for less than 3 months in an aqueous formulation containing less than about 50 mM of an alkali metal chloride salt at a pH between about 5.5 and about 7.5.
In a specific embodiment of the compositions provided above, the labile protein is stable for less than 2 months in an aqueous formulation containing less than about 50 mM of an alkali metal chloride salt at a pH between about 5.5 and about 7.5.
In a specific embodiment of the compositions provided above, the labile protein is stable for less than 1 month in an aqueous formulation containing less than about 50 mM of an alkali metal chloride salt at a pH between about 5.5 and about 7.5.
In a specific embodiment of the compositions provided above, the labile protein is stable for less than 2 weeks in an aqueous formulation containing less than about 50 mM of an alkali metal chloride salt at a pH between about 5.5 and about 7.5.
In a specific embodiment of the compositions provided above, the concentration of the labile therapeutic protein is at least about 50 mg/mL.
In a specific embodiment of the compositions provided above, the concentration of the labile therapeutic protein is at least about 100 mg/mL.
In a specific embodiment of the compositions provided above, the concentration of the labile therapeutic protein is at least about 150 mg/mL.
In a specific embodiment of the compositions provided above, the composition is formulated for subcutaneous or intramuscular administration.
In a specific embodiment of the compositions provided above, the alkali metal chloride salt is sodium chloride.
In a specific embodiment of the compositions provided above, the stabilizing agent is an amino acid.
In a specific embodiment of the compositions provided above, the amino acid is glycine.
In a specific embodiment of the compositions provided above, the concentration of the amino acid is at least about 100 mM.
In a specific embodiment of the compositions provided above, the composition is stable for at least about 3 months.
In a specific embodiment of the compositions provided above, the composition is stable for at least about 6 months.
In a specific embodiment of the compositions provided above, the composition is stable for at least about 1 year.
In a specific embodiment of the compositions provided above, the composition is stable for at least about 2 years.
In one embodiment, the present invention provides a method for stabilizing a aqueous formulation of a labile therapeutic protein at a pH between about 5.5 and about 7.5, the method comprising addition of an alkali metal chloride salt at a concentration of between about 75 mM and about 200 mM.
In one embodiment, the present invention provides a method for stabilizing a aqueous formulation of a labile therapeutic protein at a pH between about 5.5 and about 7.0, the method comprising addition of an alkali metal chloride salt at a concentration of between about 100 mM and about 200 mM.
In a specific embodiment of the methods provided above, the solution further comprises a stabilizing agent.
In a specific embodiment of the methods provided above, the labile therapeutic protein is a human or humanized protein.
In a specific embodiment of the methods provided above, the protein is a recombinant protein.
In a specific embodiment of the methods provided above, the protein is a plasma-derived protein.
In a specific embodiment of the methods provided above, the protein is an immunoglobulin.
In a specific embodiment of the methods provided above, the immunoglobulin is an IgG preparation.
In a specific embodiment of the methods provided above, the protein concentration of the IgG preparation is at least about 150 mg/mg.
In a specific embodiment of the methods provided above, the protein concentration of the IgG preparation is at least about 200 mg/mg.
In a specific embodiment of the methods provided above, the immunoglobulin is a recombinant antibody.
In a specific embodiment of the methods provided above, the method stabilizes the immunoglobulin composition for at least 6 months when stored at between about 38° C. and about 42° C.
In a specific embodiment of the methods provided above, the method stabilizes the immunoglobulin composition for at least 1 year when stored at between about 28° C. and about 32° C.
In a specific embodiment of the methods provided above, the pH of the composition is between about 5.5 and about 6.5.
In a specific embodiment of the methods provided above, the protein is a coagulation protein.
In a specific embodiment of the methods provided above, the coagulation protein is Factor VIII.
In a specific embodiment of the methods provided above, the pH of the composition is between about 6.0 and about 7.0.
In a specific embodiment of the methods provided above, the pH of the composition is 6.5±0.2.
In a specific embodiment of the methods provided above, the method stabilizes the composition such that at least 80% of the Factor VIII activity is retained after storage at a temperature between about 2° C. and about 8° C. for at least 3 months.
In a specific embodiment of the methods provided above, the coagulation protein is Factor VII.
In a specific embodiment of the methods provided above, the coagulation protein is Factor IX.
In a specific embodiment of the methods provided above, the coagulation protein is von Willebrand Factor (vWF).
In a specific embodiment of the methods provided above, the coagulation protein is a protein K-dependent coagulation complex.
In a specific embodiment of the methods provided above, the protein K-dependent coagulation complex comprises the coagulation factors Factor II, Factor IX, and Factor X.
In a specific embodiment of the methods provided above, the protein K-dependent coagulation complex further comprises Factor VII.
In a specific embodiment of the methods provided above, the labile protein is stable for less than 3 months in an aqueous formulation containing less than about 50 mM of an alkali metal chloride salt at a pH between about 5.5 and about 7.5.
In a specific embodiment of the methods provided above, the labile protein is stable for less than 2 months in an aqueous formulation containing less than about 50 mM of an alkali metal chloride salt at a pH between about 5.5 and about 7.5.
In a specific embodiment of the methods provided above, the labile protein is stable for less than 1 month in an aqueous formulation containing less than about 50 mM of an alkali metal chloride salt at a pH between about 5.5 and about 7.5.
In a specific embodiment of the methods provided above, the labile protein is stable for less than 2 weeks in an aqueous formulation containing less than about 50 mM of an alkali metal chloride salt at a pH between about 5.5 and about 7.5.
In a specific embodiment of the methods provided above, the concentration of the labile therapeutic protein is at least about 50 mg/mL.
In a specific embodiment of the methods provided above, the concentration of the labile therapeutic protein is at least about 100 mg/mL.
In a specific embodiment of the methods provided above, the concentration of the labile therapeutic protein is at least about 150 mg/mL.
In a specific embodiment of the methods provided above, the composition is formulated for subcutaneous or intramuscular administration.
In a specific embodiment of the methods provided above, the alkali metal chloride salt is sodium chloride.
In a specific embodiment of the methods provided above, the stabilizing agent is an amino acid.
In a specific embodiment of the methods provided above, the amino acid is glycine.
In a specific embodiment of the methods provided above, the concentration of the amino acid is at least about 100 mM.
In a specific embodiment of the methods provided above; the method increases the time of stability for the composition by at least 25%.
In a specific embodiment of the methods provided above, the method increases the time of stability for the composition by at least 50%.
In a specific embodiment of the methods provided above, the method increases the time of stability for the composition by at least 100%.
In a specific embodiment of the methods provided above, the method increases the time of stability for the composition by at least 200%.
To determine the role pH and salt concentration have on a plasma-derived 20% IgG composition, a two year stability study was conducted. This study revealed that the inclusion of sodium chloride and/or the formulation at neutral to mildly acid pH imparted a stabilizing effect on the 20% IgG composition.
Briefly, two IgG compositions prepared from pooled plasma according to the Gammagard SD process outlined in Teschner et al. (Vox Sang. 2007 January; 92(1):42-55) were concentrated to a final protein concentration of 20%. These preparations were then divided into several samples which were differentially formulated at pHs 6.5, 7.0, or 7.5 with and without 50 mM sodium chloride. The aqueous formulations were then stored at between 28° C. and 32° C. for 24 months. After the two year incubation period, the molecular size distributions of the IgG in the various formulations were investigated by high performance size exclusion chromatography (HP-SEC), the results of which are provided in Table 5.
The results shown in Table 5 indicate that, within the pH range of 6.5 to 7.5, increases in pH result in increased aggregation of the IgG preparation, as shown by the increasing percentage of IgG polymers in the formulations at pH 7.0 and 7.5 compared to the formulations at pH 6.5. Inclusion of 50 mM sodium chloride in each formulation stabilizes the lower molecular weight IgG species, resulting in a nearly 14% reduction in the level of IgG polymers in the samples. This is in contrast to IgG compositions formulated as low pH (4.4 to 4.9), which are destabilized by the addition of sodium chloride.
To further characterize the stabilizing effect that sodium chloride has on IgG compositions formulated at mildly acid to neutral pH, an accelerated stability study was performed. For the accelerated study, elevated temperatures (38° C. to 42° C.) were used to simulate longer time periods at room temperature (20° C. to 25° C.). Briefly, a 20% IgG composition, prepared as in Example 1, was divided into samples that were formulated with increasing salt concentrations (0 mM, 50 mM, 100 mM, and 150 mM) at mildly acid to neutral pHs (pH 5.5, 6.0, 6.5, 7.0, and 7.5). The aqueous formulations were then stored at between 38° C. and 42° C. for 6 months. After the 6 month incubation period, the molecular size distributions of the IgG in the various formulations were investigated by high performance size exclusion chromatography (HP-SEC). The percentage of IgG aggregates present in the various formulations is shown in
As seen in
To evaluate the stabilizing effects of sodium chloride on other immunoglobulin preparations formulated at mildly acidic to neutral pH, preparations of Partobulin® NG (Baxter Biosciences) and Tetabulin® NG (Baxter Biosciences) were formulated accordingly and tested for stability, via aggregation formation, and activity, via anti-antigen potency, over a 6 month time frame. Partobulin® is a plasma-derived human anti-D antigen immunoglobulin preparation used for antenatal anti-D prophylaxis in Rh(D) negative pregnant women carrying Rh(D) positive fetuses, as well as for the treatment of Rh(D) negative persons after incompatible transfusions of Rh(D) positive blood or erythrocyte concentrate. Tetabulin® is a plasma-derived human tetanus immunoglobulin used for post-tetanus exposure prophylaxis and therapy of clinically manifest tetanus. Both Partobulin® and Tetabulin® are typically formulated at between 100 g/L and 170 g/L human protein (of which at least 90% are immunoglobulin G) for intramuscular administration.
Briefly, feasibility lots of Partobulin® NG and Tetabulin® NG were stored at low pH for one year at 2° C. to 8° C. prior to the addition of sodium chloride to 150 mM and adjustment of the pH to between 5.5 and 7.5. Prior to the formulation with sodium chloride and pH adjustment, the bulk was formulated at pH 4.7 in the presence of 250 mM glycine like for GGL/KIOVIG. The stability of the formulations at 28° C. to 32° C. was then tracked for 6 months by periodically monitoring the Tetanus anti-toxin or Anti-D titers as well as the molecular size distributions of the formulations.
Aggregate formation over the course of 6 months for Tetabulin® NG and Partobulin® NG are shown in
Despite the increased aggregation seen in the Tetabulin® NG formulations at pH 7.0 and 7.5, no significant loss in Tetanus anti-toxin titer was found over the course of the six month storage period (
To determine if the stabilizing effect of sodium chloride at mildly acidic to neutral pH was specific to plasma-derived immunoglobulin preparations or broadly applicable to all immunoglobulin products, the stability of a recombinant anti-MIF antibody was determined in an accelerated stability assay. Briefly, samples of bulk recombinant anti-MIF antibody at 120 g/L were formulated with 150 mM sodium chloride and 0.25 M glycine at pH 4.5, 5.6, 6.5, and 7.3 and stored at between 38° C. and 42° C. for three months.
The molecular size distributions of the IgG in the various formulations were determined by high performance size exclusion chromatography (HPLC-SEC) at the start of the storage period, after two weeks, 1 month, and 3 months of storage. The results of these characterizations are provided in Table 6.
As seen in Table 6, fragmentation of the recombinant anti-MIF antibody is most pronounced at acidic pH (pH 4.5), even after only 2 weeks storage. Likewise, as seen for the concentrated IgG preparation in Example 1 and Tetabulin® and Partobulin® formulations in Example 3, sodium chloride reduced aggregation of the recombinant anti-MIF antibodies at mildly acid conditions (pH 5.6 and 6.5), while aggregation occurred more quickly at neutral pH (pH 7.5).
To further characterize the stabilizing effects of sodium chloride in these formulations, anti-MIF titers were determined by ELISA for each formulation at the start of the storage period, and after 2 weeks, 1 month, and 3 months of storage at elevated temperatures (38° C. to 42° C.). As seen in
Factor VIII, a coagulation factor, is a highly labile therapeutic protein that is commonly formulated as a lyophilized powder that must be reconstituted prior to administration. In order to determine if the addition of sodium chloride can also stabilize non-immunoglobulin labile protein formulations at mildly acidic to neutral pH, stability tests were performed on recombinant Factor VIII formulations. Briefly, frozen recombinant Factor VIII was slowly thawed and formulated by buffer exchange according to the reconstituted formulation for ADVATE (recombinant FVIII; Baxter Healthcare Corporation, Westlake Village, Calif.), given in Table 7. Samples of the FVIII preparation were then aliquoted and the pH adjusted to pH 5.5, 6.0, 6.5, 7.0, 7.5, or 8.0 with and without addition of 150 mM sodium chloride to give formulations with a starting Factor VIII activity in solution of 500 IU/mL. The samples were stored at 2° C. to 8° C. for 12 weeks, during which time the residual FVIII activity was periodically monitored.
In the absence of sodium chloride in the formulation, a continuous drop in FVIII activity was seen for all formulation (
Conversely, addition of sodium chloride in the rFVIII formulations resulted in a marked increase in the stability of FVIII activity at pH above 6.0 (
The effect sodium chloride has on the stability of an IgG composition formulated at acidic pH (4.4 to 4.9) was investigated. Briefly, an IgG composition was formulated with 0.25 M glycine, pH 4.4 to 4.9, at a final protein concentration of 10%. Sodium chloride was then added to aliquots of the formulation to a final concentration of 0 mM, 10 mM, 50 mM, or 100 mM. The molecular size distribution was then monitored by HPLC analysis after 1, 3, 6, 9, 12, 19, and 24 months of storage under refrigerated conditions (between 2° C. and 8° C.). The fixation of complement in the samples was also measured to determine the tolerability of the formulation. The results of the analysis are given in Table 8.
As can be seen in Table 8, Storage of the 10% IgG preparation at between 2° C. and 8° C., with different amounts of sodium chloride added, revealed that the presence of sodium chloride seriously impacts the ACA value and therefore the tolerability of the product, as well as the oligo/dimmer content of the samples. Compared to the final container without sodium chloride, which meets the required specifications for the product over the entire storage time, a content of 10 mM sodium chloride is sufficient to reduce the storage period to 19 months, a content of 50 mM to 12 months and a concentration of 100 mM to 9 months.
To confirm the negative effect sodium chloride has on the stability of immunoglobulin compositions formulated at acidic pH, the experiment was repeated with storage at room temperature (28° C. to 30° C.). As can be seen in Table 9, the results of the second experiment confirm the data obtained for storage between 2° C. and 8° C. The ACA values are generally lower, while the fragmentation and aggregation is accelerated. The negative effect of the addition of sodium chloride at acidic pH is confirmed with this experiment.
It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.
The present application claims the benefit of U.S. Provisional Application No. 61/384,209, filed Sep. 17, 2010, the content of which is expressly incorporated herein by reference in its entirety for all purposes.
| Number | Date | Country | |
|---|---|---|---|
| 61384209 | Sep 2010 | US |
