Formulations and uses thereof
AssigneeSartorius Albumedix Limited
InventorsPhilip Harvey Morton, Neil Dodsworth, Eleonora Cerasoli, Jason Cameron
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Abstract
The invention provides formulations of albumin with desirably high levels of free thiol and desirably low levels of albumin polymer. The invention further provides uses of a formulation comprising: (i) albumin, wherein the albumin is in a substantially unaggregated form; and (ii) an anion having multiple negative charges housed on a flexible backbone, to stabilise a virus. The invention also provides a formulation and a kit of parts comprising (i) albumin, wherein the albumin is in a substantially unaggregated form; and (ii) an anion having multiple negative charges housed on a flexible backbone and its use to stabilise a virus.
Claims182
-167. (canceled)
Use of a liquid formulation comprising:
- (i) albumin;
- and
- (ii) an anion having multiple negative charges housed on a flexible backbone, to stabilise a virus, wherein the albumin in the formulation is one that meets the following parameters:
- the polydispersity index of albumin as assessed by dynamic light scattering is equal to or less than 0.05;
- and
- less than 0.025% (w/w) of albumin in the formulation is in a polymeric form of higher order aggregates or multimers excluding dimers and trimers;
- and
- wherein the stabilising of the virus is manifest by decreased aggregation of the virus when subjected to stress conditions selected from freeze-thaw cycles or incubating at a set temperature for a set period of time, compared to the level of aggregation of the virus in the absence of the formulation but otherwise under the same conditions.
The use according to any one of the preceding claims, wherein the amount of albumin in a polymeric form is assessed by high-performance liquid size exclusion chromatography (SEC-HPLC).
The use according to any one of the preceding claims, wherein the albumin polymer level and polydispersity index parameters are observed within 6 months of formulation.
The use according to any one of the preceding claims, wherein the anion having multiple negative charges housed on a flexible backbone is not one of or both of glutamate and EDTA.
The use according to any one of the preceding claims, wherein the anion having multiple negative charges housed on a flexible backbone is not one of or both of glutamate and ATP.
The use according to any one of the preceding claims, wherein the anion having multiple negative charges housed on a flexible backbone is a multivalent carboxylic acid such as citrate, fumarate, tartaric acid, α-ketoglutarate, malate, maleic acid, succinate, succinic acid, aconitate, isocitrate, oxaloacetate, adenosine triphosphate or sodium tripolyphosphate or any intermediate anion of the citric acid cycle.
The use according to any one of the preceding claims, wherein the formulation is substantially free of a sugar and/or a sugar alcohol.
The use according to any one of the preceding claims, wherein the amount of albumin in a polymeric form is as assessed by high-performance liquid size exclusion chromatography (SEC-HPLC) using an injection sample of albumin provided at or diluted to less than 100 mg/mL.
The use according to any one of the preceding claims, wherein the albumin is a recombinant albumin.
The use according to any one of the preceding claims, wherein the formulation comprises at least 0.5 moles free thiol per mole albumin.
The use according to any one of the preceding claims, wherein the albumin is provided by an albumin composition selected from the group consisting of any of:
- a. a composition comprising 130 to 160 mM, preferably 145 mM, of sodium, 29 to 35 mM, preferably 32 mM, of octanoate, 10 to 20 mg/L, preferably 10 to 15 mg/L, of Polysorbate 80, 190 to 210 mg/mL, preferably 200 mg/mL, of albumin (w/v), wherein the pH is 6.7 to 7.3, preferably 7 and wherein the percentage of albumin (w/w) in a polymeric form is 0 to 1%, more preferably less than 0.025%;
- b. a composition comprising 120 to 160 mM, preferably 145 mM, of sodium, 4 to 12 mM, preferably 8 mM, of octanoate, 0 to 50 mg/L, preferably 25 to 45 mg/L, of Polysorbate 80, 95 to 105 mg/mL, preferably 100 mg/mL, of albumin (w/v), wherein the pH is 6.4 to 7.4, preferably 7, and wherein the percentage of albumin (w/w) in a polymeric form is 0 to 1%, more preferably less than 0.025%;
- c. a composition comprising 120 to 160 mM, preferably 145 mM, of sodium, 8 to 24 mM, preferably 16 mM, of octanoate, 0 to 100 mg/L, preferably 50 to 70 mg/L, of Polysorbate 80, 190 to 210 mg/mL, preferably 200 mg/mL, of albumin (w/v), wherein the pH is 6.4 to 7.4, preferably 7, and wherein the percentage of albumin (w/w) in a polymeric form is 0 to 1%, more preferably less than 0.025%;
- d. a composition comprising 200 to 300 mM, preferably 230 to 260 mM of sodium, 0 to 3 mM, preferably 0 to 1 mM of octanoate, 95 to 105 mg/mL, preferably 100 mg/mL, of albumin (w/v), wherein the pH is 6 to 7, preferably 6.5, and wherein the percentage of albumin (w/w) in a polymeric form is 0 to 1%, preferably less than 0.025%.
The use according to any one of the preceding claims, wherein the formulation comprises from 0.4 to 300 mg/mL albumin, such as 0.4-50 mg/mL of albumin, optionally wherein the formulation comprises 1-20 mg/mL of albumin.
The use according to any one of the preceding claims, wherein the anion having multiple negative charges housed on a flexible backbone is one that thermally stabilises the albumin.
The use according to any one of the preceding claims, wherein the anion having multiple negative charges housed on a flexible backbone protects against deamidation of the albumin.
The use according to any one of the preceding claims, wherein the anion having multiple negative charges housed on a flexible backbone protects against N-terminal degradation of the albumin.
The use according to any one of the preceding claims, wherein the anion having multiple negative charges housed on a flexible backbone is present at a concentration of 2-500 mM or 10-500 mM, optionally wherein the anion is present at a concentration of 2-100 mM or 25-100 mM.
The use according to any one of the preceding claims, wherein the formulation further comprises a buffer.
The use according to any one of the preceding claims, wherein the formulation comprises 0-50 mM of magnesium chloride, optionally wherein the formulation comprises about 25 mM of magnesium chloride or about 5 to about 10 mM magnesium chloride, such as about 5 mM or about 10 mM magnesium chloride.
The use according to any one of the preceding claims, wherein:
- (i) the formulation comprises 0-500 mM, such as 0-200 mM of sulfate salts, optionally wherein the formulation comprises about 50-500 mM of sulfate salts, such as about 50-200 mM, such as about 50 mM of sulfate salts or about 200 mM of sulfate salts, and further optionally wherein the sulfate salt is sodium sulfate, sodium bisulfate, sodium thiosulfate, ammonium sulfate, ammonium bisulfate, magnesium sulfate, magnesium bisulfate, potassium sulfate, potassium bisulfate;
- and/or
- (ii) the formulation comprises 0-500 mM of disodium phosphate, such as 0-200 mM optionally wherein the formulation comprises about 10 mM of disodium phosphate or about 200 mM of disodium phosphate;
- and/or
- (iii) the formulation comprises 0-50 mM of EDTA, optionally wherein the formulation comprises about 1 mM of EDTA;
- and/or
- (iv) the formulation comprises 0-200 mM of glutamic acid, optionally wherein the formulation comprises about 50 mM of glutamic acid;
- and/or
- (v) the formulation comprises 0-500 mM of mannitol, optionally wherein the formulation comprises about 300 mM of mannitol;
- and/or
- (vi) the formulation comprises 0-500 mM of sucrose, optionally wherein the formulation comprises about 300 mM of sucrose;
- and/or
- (vii) the formulation comprises 0-500 mM of sodium chloride, such as about 0-200 mM, optionally wherein the formulation comprises about 50-200 mM of sodium chloride, such as about 200 mM or about 50 mM sodium chloride.
The use according to any one of the preceding claims, wherein the formulation comprises 0-500 mM of arginine, such as about 0-200 mM, optionally wherein the formulation comprises about 25 mM to about 50 mM of arginine.
The use according to any one of the preceding claims, wherein the formulation comprises 0-500 mM such as 0-200 mM of chloride salts, optionally wherein the formulation comprises about 50 mM of chloride salts.
The use according to any one of the preceding claims, wherein the formulation comprises 0-200 mM of histidine, optionally wherein the formulation comprises about 50 mM of histidine.
The use according to any one of the preceding claims, wherein the formulation comprises 0-200 mM of glycine, optionally wherein the formulation comprises about 50 mM of glycine or about 200 mM of glycine.
The use according to any one of the preceding claims, wherein the formulation comprises 0-200 mM of glutamate, optionally wherein the formulation comprises about 50 mM of glutamate.
The use according to any one of the preceding claims, wherein the formulation comprises 0-200 mM of aspartate, optionally wherein the formulation comprises about 50 mM of aspartate.
The use according to any one of the preceding claims, wherein the formulation comprises 0-0.01% (w/v) of non-ionic surfactant, optionally wherein the formulation comprises about 0.001% or about 0.01% (w/v) of non-ionic surfactant, further optionally wherein the non-ionic surfactant is a poloxamer such as Pluronic F-68 or a polysorbate.
The use according to any one of the preceding claims, wherein the pH of the formulation is from about 4 to about 8, optionally wherein the pH of the formulation is from about 5 to about 7.5, preferably wherein the pH of the formulation is about 6 or about 6.5.
The use according to any one of the preceding claims, wherein the increased stability is manifest by the coefficient of variance of the hydrodynamic radius of the formulation containing the virus being not more than 5% when the formulation is incubated at 40° C. for a duration of at least 20 hours.
The use according to any one of the preceding claims, wherein the increased stability is manifest by a particle count of less than 60 000 particles measured by backgrounded membrane imaging (BMI) at after storage or incubation at a temperature of from 2 to 8° C., such as from 2, 3, 4, 5, 6 or 7 to 3, 4, 5, 6, 7 or 8° C., for a duration of at least 1 month, for example, at least 2, 3, 4, 5 or 6 months, wherein the estimated circular diameter (ECD) of the particle is equal to or less than 2 μm (microns).
The use according to any one of the preceding claims, wherein the increased stability is manifest by a particle count of less than 15 000 particles measured by backgrounded membrane imaging (BMI) after storage or incubation at a temperature of from 2 to 8° C. such as from 2, 3, 4, 5, 6 or 7 to 3, 4, 5, 6, 7 or 8° C., for example at about 4° C., for a duration of at least 1 month, for example, at least 2, 3, 4, 5 or 6 months, wherein the estimated circular diameter (ECD) of the particle is equal to or less than 2 μm (microns) and the particles are Thioflavin T-positive.
The use according to any one of the preceding claims, wherein the increased stability is manifest by a particle count of less than 200 000 particles in Brightfield-Side illumination membrane intensity (SIMI) measured by backgrounded membrane imaging after at least four rounds of freeze-thaw cycles, wherein the estimated circular diameter (ECD) of the particle is equal to or less than 2 μm (microns).
The use according to any one of the preceding claims, wherein the increased stability is manifest by the hydrodynamic radius of the formulation containing the virus being no more than 6× (preferably no more than 5.5×, 5×, 4.5×, 4×, 3.5×, 3×, 2.5×, 2×, 1.5×) the hydrodynamic radius of the virus in its monomeric form, after freeze-thaw stress (e.g. following 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 cycles of storing at a temperature of around −20° C. for at least 6 hours, followed by thawing at room temperature for about 3 hours).
The use according to any one of the preceding claims, wherein the increased stability is manifest by a mass percentage recovery for the virus of at least 50%, such as at least 55%, 60%, 65%, 70%, 75%, 80%, 85% 90%, 95%, 98% or 99%, after freeze-thaw stress (e.g. following 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 cycles of storing at a temperature of around −20° C. for at least 6 hours, followed by thawing at room temperature for about 3 hours).
The use according to any one of the preceding claims, wherein the increased stability is manifest by the mass percentage recovery for the virus being increased or improved by at least 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 percentage points relative to the mass percentage recovery of the virus in the absence of the formulation (e.g. the virus in an otherwise identical or substantially identical formulation but which contains no albumin and/or no anion having multiple negative charges housed on a flexible backbone), after freeze thaw stress (e.g. following 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 cycles of storing at a temperature of around −20° C. for at least 6 hours, followed by thawing at room temperature for about 3 hours.
The use according to any one of the preceding claims, wherein the increased stability is manifest by the particle count of the formulation containing the virus being less than the particle count of a control solution of the virus (e.g. a solution of the virus in the absence of the albumin and/or in the absence of the anion having multiple negative charges housed on a flexible backbone) after freeze thaw stress (e.g. following 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 cycles of storing at a temperature of around −20° C. for at least 6 hours, followed by thawing at room temperature for about 3 hours.
The use according to any one of the preceding claims, wherein the increased stability is manifest by retention of virus integrity.
The use according to any one of the preceding claims, wherein the increased stability is manifest by retention of the ability of the virus to bind to a specific binding partner of the virus.
The use according to any one of the preceding claims, wherein the increased stability is manifest by retention of the ability of the virus to infect a cell.
The use according to any one of the preceding claims, wherein the virus is (i) any of a viral particle, a viral fragment, a virus-like particle, a viral vector or a vector virus and/or (ii) a simian virus, optionally wherein the simian virus is an adeno-associated virus (AAV), a recombinant AAV, Adenovirus, Herpes Simplex Virus, vesicular stomatitis virus, Maraba virus, Vaccinia virus, Measles virus, Newcastle Disease Virus, Rotavirus, Reovirus, Type I Poliovirus, Coxsackievirus A21, Seneca Valley virus, Dengue virus, Yellow fever virus, West Nile virus, zika virus, Alphavirus or Coronavirus.
The use according to any one of the preceding claims, wherein the virus is a live virus, an attenuated virus, a live-attenuated virus or an inactivated virus.
The use according to any one of the preceding claims, wherein the virus is in the form of a vaccine.
The use according to any one of the preceding claims, wherein the virus is a hybrid or chimeric virus.
The use according to any one of the preceding claims, wherein the virus that is stabilised is at a concentration of 1×106-1×1024 vg/mL.
A formulation comprising:
- (i) albumin;
- and
- (ii) an anion having multiple negative charges housed on a flexible backbone, wherein the albumin in the formulation is one that meets the following parameters:
- the polydispersity index of albumin as assessed by dynamic light scattering is equal to or less than 0.05;
- and
- less than 0.025% (w/w) of albumin is in a polymeric form of higher order aggregates or multimers excluding dimers and trimers.
A kit of parts comprising:
- (i) albumin;
- and
- (ii) an anion having multiple negative charges housed on a flexible backbone, wherein the albumin in the kit is one that meets the following parameters:
- the polydispersity index of albumin as assessed by dynamic light scattering is equal to or less than 0.05;
- and
- less than 0.025% (w/w) of albumin is in a polymeric form of higher order aggregates or multimers excluding dimers or trimers.
A method of selecting a suitable albumin for use in stabilising a virus, the method comprising determining whether the albumin meets the following parameters:
- the polydispersity index of albumin as assessed by dynamic light scattering is equal to or less than 0.05;
- and
- less than 0.025% (w/w) of albumin is in a polymeric form of higher order aggregates or multimers excluding dimers and trimers;
- and optionally—the hydrodynamic radius of albumin does not change more than 4.5% when incubated at 40° C. for a duration of 3 days;
- wherein an albumin that meets the parameters is one that is useful in stabilising a virus, wherein the stabilising of the virus is manifest by decreased aggregation of the virus when subjected to stress conditions selected from freeze-thaw cycles or incubating at a set temperature for a set period of time.
A liquid formulation of albumin or fragment thereof comprising at least 0.75 moles free thiol per mole albumin or fragment thereof and at most 1% (w/w) albumin polymer after storage or incubation at at least 40° C. for at least 1, 2, 3, 4, 5 or 6 months.
A liquid formulation of albumin or fragment thereof comprising at least 0.75 moles free thiol per mole albumin or fragment thereof and at most 1% (w/w) albumin polymer after storage or incubation at at least 25° C. for at least 1, 2, 3, 4, 5, 6, 9 or 12 months.
A liquid formulation of albumin or fragment thereof comprising at least 0.75 moles free thiol per mole albumin and at most 1% (w/w) albumin polymer or fragment thereof after storage or incubation at a temperature of from 2 to 8° C., such as from 2, 3, 4, 5, 6, or 7 to 3, 4, 5, 6, 7, or 8° C., for example at about 4° C., for a duration of at least 1, 2, 3, 4, 5 or 6 months or at least 1, 2, 3, 4, 5 or 6 years.
A liquid formulation comprising albumin, fatty acid and cation, in which the formulation lies within a design space defined by both of the following equations:
- Free
-
- thiol
- =
- 1.1786
- -
- 0.05167
- *
- pH
- -
- 0.0001544
- *
- cation
- -
- 0.01133
- *
- FA
- +
- (
- pH
- -
- 6
- )
- *
- (
- (
- cation
- -
- 198.3
- )
- *
- 0.0002659
- )
- +
- (
- pH
- -
- 6
- )
- *
- (
- (
- FA
- -
- 7.125
- )
- *
- 0.00316
- )
- +
- (
- cation
- -
- 198.3
- )
- *
- (
- (
- FA
- -
- 7.125
- )
- *
- 0.000001935
- )
- +
- (
- pH
- -
- 6
- )
- *
- (
- (
- pH
- -
- 6
- )
- *
- -
- 0.05
- )
- +
- (
- cation
- -
- 198.3
- )
- *
- (
- (
- cation
- -
- 198.3
- )
- *
- 0.000002583
- )
- +
- (
- FA
- -
- 7.125
- )
- *
- (
- (
- FA
- -
- 7.125
- )
- *
- 0.0001413
- )
- +
- (
- pH
- -
- 6
- )
- *
- (
- (
- cation
- -
- 198.3
- )
- *
- (
- (
- FA
- -
- 7.125
- )
- *
- -
- 0.000009879
- )
- )
- ,
- and
- (
- a
- )
- Polymer
- =
- 9.1677
- -
- 0.8417
- *
- pH
- -
- 0.01044
- *
- cation
- -
- 0.269
- *
- FA
- +
- (
- pH
- -
- 6
- )
- *
- (
- (
- cation
- -
- 198.3
- )
- *
- -
- 0.004796
- )
- +
- (
- pH
- -
- 6
- )
- *
- (
- (
- FA
- -
- 7.125
- )
- *
- 0.06523
- )
- +
- (
- cation
- -
- 198.3
- )
- *
- (
- (
- FA
- -
- 7.125
- )
- *
- 0.001494
- )
- +
- (
- pH
- -
- 6
- )
- *
- (
- (
- pH
- -
- 6
- )
- *
- 2.25
- )
- +
- (
- cation
- -
- 198.3
- )
- *
- (
- (
- cation
- -
- 198.3
- )
- *
- 0.00006768
- )
- +
- (
- FA
- -
- 7.125
- )
- *
- (
- (
- FA
- -
- 7.125
- )
- *
- 0.0249
- )
- +
- (
- pH
- -
- 6
- )
- *
- (
- (
- cation
- -
- 198.3
- )
- *
- (
- (
- FA
- -
- 7.125
- )
- *
- 0.001237
- )
- )
- ;
- (
- b
- )
- where
- ‘free thiol’ means the level of free thiol,
- ‘cation’ means the cation concentration in mM,
- ‘FA’ means the fatty acid concentration in mM.
Dependencies are inferred from the claim text.