EP1685160A1 - Dry recombinant human alpha 1-antitrypsin formulation - Google Patents
Dry recombinant human alpha 1-antitrypsin formulationInfo
- Publication number
- EP1685160A1 EP1685160A1 EP04798463A EP04798463A EP1685160A1 EP 1685160 A1 EP1685160 A1 EP 1685160A1 EP 04798463 A EP04798463 A EP 04798463A EP 04798463 A EP04798463 A EP 04798463A EP 1685160 A1 EP1685160 A1 EP 1685160A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- dry powder
- powder composition
- raat
- less
- protein content
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/81—Protease inhibitors
- C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
- C07K14/811—Serine protease (E.C. 3.4.21) inhibitors
- C07K14/8121—Serpins
- C07K14/8125—Alpha-1-antitrypsin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/55—Protease inhibitors
- A61K38/57—Protease inhibitors from animals; from humans
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1611—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
Definitions
- This invention relates to a dry protein formulation, and in particular to a formulation of alphal-antitrypsin (AAT).
- AAT alphal-antitrypsin
- Background of the Invention AAT and recombinant alphal -antitrypsin (rAAT) are potential therapeutic agents for a number of clinical indications.
- rAAT is a 395 amino acid protein of 44 kD, that is non-glycosylated and has an amino acid sequence identical to the human plasma protein (AAT) with the exception of an N-acetylmethionine residue at the amino terminus. It is desirable to have a dry, stable formulation of AAT or rAAT, ready for reconstitution in water and immediate use.
- Excipients typically employed in a dried protein formulation comprise mainly buffers, sugars and surfactants.
- Other potential stabilizing excipients include bulking agents, chelating agents, antioxidants, reducing agents and amino-acids.
- US5780014A describes a dry powder formulation of AAT, for administration by inhalation. Various drying techniques are suggested.
- Prolastin (Bayer) is a lyophilized preparation of human plasma-derived, glycosylated AAT.
- the liquid composition When reconstituted as directed, at 1 g alphal-antitrypsin functional activity per 40 mL sterile water, the liquid composition comprises >20 mg/ml AAT, 100-210 mEq/L Na, 60-180 mEq/L CI, 15-25 ⁇ sodium phosphate, ⁇ 5 ppm PEG and ⁇ 0.1 % sucrose.
- the lyophilized formulation should be stored under refrigeration.
- Vemuri et al in Chapter 9 of Stability and Characterization of Protein and Peptide Drugs: Case Histories, ed. Wang and Pearlman, Plenum Press, New York (1993), describe formulations of rAAT, primarily in liquid form. Stability, e.g. at pH 7.5, is enhanced by increasing the salt content.
- salt is generally considered unsuitable for a lyophilized formulation because of the reduced glass transition temperature.
- the stabilization of rAAT presents particular problems, relative to the natural protein. Travis et al., (J. Biol. Chem. 260:4384-4389,1985) describe a comparison of heat stabilities of yeast-derived rAAT with natural plasma-derived AAT. The half-life of non-glycosylated rAAT, with respect to its activity in response to thermal stress, is considerably less than that of its natural glycosylated counterpart. Summary of the Invention The present invention is based on the discovery of a dry formulation of rAAT, having defined concentrations of rAAT and salt, that has good stability, even without refrigeration (i.e.
- Figure 1 is a FTIR spectral scan of liquid and solid rAAT in a formulation of the invention.
- Figure 2 is a FTIR spectral scan of unformulated rAAT in the liquid and solid states.
- Figure 3 is a FTIR spectral scan of rAAT in formulations containing different levels of salt.
- Figure 4 shows the secondary structure of rAAT in a sugar-based formulation and in a salt-based formulation of the invention.
- Figure 5 illustrates the reversibility of the secondary structure of rAAT to its original structure upon reconstitution of the lyophilized protein in 100 mM NaCI formulation.
- Description of Preferred Embodiments A dried formulation according to the invention contains at least rAAT and salt. Although their effect on the stability of the composition is relatively small, other, conventional components may be included. Such components include reducing agents such as dithiothreitol, cysteine, glutathione, or N-acetylcysteine
- the composition may also contain antioxidants, such as ascorbic acid or L-Met, e.g. in an amount of up to 10 mM on reconstitution and/or a buffer such as phosphate, citrate or histidine, e.g. in an amount of 5-50 mM, preferably 10-20 mM on reconstitution.
- the amount of buffer may be such that, on reconstruction of the composition in water, the reconstituted solution has a pH of from about 6 to 9, more preferably 6.5-8, preferably from 6.8-70.
- Othertypical constituents are chelating agents (e.g. EDTA or citrate), and surfactants (e.g., polyoxyethylene sorbitan).
- the dry powder composition of the invention does not require to have been subjected to viral inactivation. That is typically done by heating, at 60°C or 65°C.
- the dry powder composition of the invention typically has a protein content which is less than 10%, more preferably less than 5%, most preferably less than 1%, ⁇ 1-antichymotrypsin.
- the composition also typically has protein content which is less than 10%, more preferably less than 5%, most preferably less than 1 %, albumin. More generally, protein content is usually less than 10%, more preferably less than 5%, most preferably less than 1 % human protein.
- the protein content is usually more than 90%, preferably more than 95% rAAT, and most preferably more than 99% rAAT.
- the dry powder composition may further comprise 1 to 2000 milliequivalents salt per 100 mg of rAAT, more preferably 50-500 milliequivalents, most preferably 100-200 milliequivalents.
- the salt that is used will typically be NaCI. However, it will be readily appreciated by those of ordinary skill in the art that other salts may have the same effect, whether the cation is different (as in KCI) or the anion is different (as in NaBr) or both.
- the dry powder composition of the invention can be free of sugar. It usually contains less than 1 % and preferably less than 0.5% water.
- the dry powder composition of the invention can retain at least 80% of initial rAAT activity, preferably > 90%, upon storage at under conditions that are, or are equivalent to, 50°C for 3 months.
- the composition may also retain at least 80% monomeric rAAT, preferably > 95% monomer, upon storage under conditions that are, or are equivalent to, 50°C for 3 months. Criteria for stability (retained activity) and denaturation are demonstrated by assays known to those skilled in the art. Activity assays are based on the porcine pancreatic elastase inhibition assay reported by Beatty et al, J Biol. Chem. 255, p. 3931 , 1980.
- Denaturation is monitored by evaluation of aggregate formation, and the non-denatured rAAT reported as % monomer, in a size exclusion chromatography (SEC) HPLC method. Equivalence to the given conditions will be understood by one of ordinary skill in the art, i.e. based on the Arrhenius equation.
- a solution or other composition comprising the desired components is dried. Suitable methods of drying include, but are not limited to, lyophilization, spray-drying, spray freeze- drying, fluidized bed technology and super critical fluid drying. Preferred drying procedures are lyophilization and spray-drying. Both procedures can be performed by standard technology known to those of ordinary skill in the art.
- spray-drying consists of a three-step process which results in dry particle formation.
- the process begins by atomizing a liquid feed into a spray of fine droplets using compressed air, followed by heating media in order to dry the droplets by evaporating the moisture content of the droplets.
- the final particles in the form of dry powder are collected as product.
- the gas and the excess fine dust are exhausted.
- These steps are carried out using three components: the atomizer in shape of a nozzle; the drying chamber; and the collecting system known as cyclone and pot.
- the dry formulation or, after reconstitution, the liquid composition is suitable for administration to a patient in need thereof. Suitable routes of administrations include, but are not limited to, inhalation, topical, sub-cutaneous and intravenous delivery.
- Suitable routes of administrations include, but are not limited to, inhalation, topical, sub-cutaneous and intravenous delivery. The following Examples illustrate the invention.
- Figure 1 shows the FTIR of liquid and solid rAAT in Formulation 917-1. Note that the amide I region (1700-1600 cm “1 ) is sensitive to changes in secondary structure and that ail peaks in the second derivative spectra are negative. Each peak in the amide I region corresponds to a different secondary structural type. There are clearly perturbations of the rAAT conformation before and after lyophilization.
- the peak near 1655 cm “1 corresponds to ⁇ -helical structure
- the band near 1635 cm “1 corresponds to ⁇ -sheet structure
- the 1688 cm “1 peak arises from extended ⁇ -strands or ⁇ -sheets.
- Random coil structure is assigned to bands near 1644 cm “1 .
- the liquid sample representing the native conformation, displays a significant amount of ⁇ -sheet and ⁇ -helical structure.
- the ⁇ -helix band is almost completely lost, while there are marked increases in bands above 1680 cm "1 , corresponding to extended and loop structures.
- Figure 3 shows the effect of salt on rAAT structure in the solid state.
- Formulations 917-1 , -3 and -4 contain 175 mM, 100 mM and 50 mM NaC
- Formulations 3 and 4 which have the lower salt concentrations, appear to have the greatest degree of structural perturbation and all three formulations are less perturbed than when no stabilizers are present. Overall, it appears that lyophilization produces some structural perturbation compared to the native conformation. The extent of the changes is minimized by the addition of excipients, including salt. It appears that a NaCI concentration above 50 mM produces a more native-like structure, with a 50-100 mM optimum.
- FIG. 4 shows the FTIR spectra of rAAT formulated in a sugar-based formulation (1008-1 ) and in a salt-based formulation (1008-2).
- the secondary structure of rAAT in both these formulations is superimposable.
- the fact that salt can accomplish the same degree of stabilization with protein at high concentrations is remarkable and not obvious.
- the original rAAT secondary structure is retained as shown in Figure 5.
- the lyophilized formulations were evaluated for short-term stability (at 1 and 3 months) under accelerated storage conditions at 60°C. It should be noted that this storage temperature is particularly harsh for evaluating protein stability and may bias the results towards the trehalose-based formulations that have a particularly high glass-transition temperature (T g ). The rationale for choosing this temperature was based on previous stability studies that assessed rAAT stability over shorter time frames. The activity and percent monomer recovered were determined for up to 3 months storage at 60°C, as shown in Tables III and IV, respectively. Table III: Specific Activity of rAAT (lU/m ⁇
- Example 2 Spray Drying Recombinant alpha 1-antitrypsin (rAAT) was spray-dried in various formulations and conditions. The activity of the resulting dry powder was assayed to evaluate the rAAT potency after drying. Table VII presents the formulations and Table VIII presents the data from these experiments. Table VII
Abstract
Description
Claims
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US51880303P | 2003-11-10 | 2003-11-10 | |
US51994603P | 2003-11-14 | 2003-11-14 | |
PCT/GB2004/004740 WO2005047323A1 (en) | 2003-11-10 | 2004-11-10 | Dry recombinant human alpha 1-antitrypsin formulation |
Publications (1)
Publication Number | Publication Date |
---|---|
EP1685160A1 true EP1685160A1 (en) | 2006-08-02 |
Family
ID=34594925
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP04798463A Withdrawn EP1685160A1 (en) | 2003-11-10 | 2004-11-10 | Dry recombinant human alpha 1-antitrypsin formulation |
Country Status (5)
Country | Link |
---|---|
EP (1) | EP1685160A1 (en) |
JP (1) | JP2007534633A (en) |
AU (1) | AU2004288854B2 (en) |
CA (1) | CA2545458A1 (en) |
WO (1) | WO2005047323A1 (en) |
Families Citing this family (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
SI1684719T1 (en) * | 2003-11-14 | 2012-07-31 | Baxter Int | Alpha 1-antitrypsin compositions and treatment methods using such compositions |
GB0710529D0 (en) | 2007-06-01 | 2007-07-11 | Circassia Ltd | Vaccine |
CN101820907B (en) | 2007-08-15 | 2013-05-01 | 切尔卡西亚有限公司 | Peptide with multiple epitopes |
WO2009059082A2 (en) * | 2007-11-02 | 2009-05-07 | Talecris Biotherapeutics, Inc. | Method, composition, and article of manufacture for providing alpha-1 antitrypsin |
GB0821806D0 (en) | 2008-11-28 | 2009-01-07 | Circassia Ltd | Compositions with reduced dimer formation |
WO2010112547A1 (en) * | 2009-03-31 | 2010-10-07 | Gambro Lundia Ab | Dialysis precursor composition |
US9353165B2 (en) | 2012-07-25 | 2016-05-31 | Grifols, S.A. | Purification of cell culture derived alpha1 protease inhibitor |
CN114848614A (en) * | 2013-07-18 | 2022-08-05 | 曼金德公司 | Heat stable dry powder pharmaceutical compositions and methods |
IL267923B2 (en) * | 2018-08-02 | 2023-06-01 | Grifols Worldwide Operations Ltd | Composition comprising highly-concentrated alpha-1 proteinase inhibitor and method for obtaining thereof |
Family Cites Families (16)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU577259B2 (en) * | 1982-08-13 | 1988-09-22 | Zymogenetics Inc. | Glycolytic promters for regulated protein expression protease inhibitor |
CA1329760C (en) * | 1987-10-29 | 1994-05-24 | Ted C. K. Lee | Plasma and recombinant protein formulations in high ionic strength media |
WO1989009784A1 (en) * | 1988-04-08 | 1989-10-19 | Commonwealth Serum Laboratories Commission | Production of heat-stable factor viii concentrate |
US6258341B1 (en) * | 1995-04-14 | 2001-07-10 | Inhale Therapeutic Systems, Inc. | Stable glassy state powder formulations |
US5780014A (en) * | 1995-04-14 | 1998-07-14 | Inhale Therapeutic Systems | Method and apparatus for pulmonary administration of dry powder alpha 1-antitrypsin |
US5763401A (en) * | 1996-07-12 | 1998-06-09 | Bayer Corporation | Stabilized albumin-free recombinant factor VIII preparation having a low sugar content |
KR100548107B1 (en) * | 1997-01-30 | 2006-02-02 | 바이오팜 게젤샤프트 쭈어 바이오테히놀로지셴 엔트비클룽 폰 파르마카 엠베하 | Freeze-Dried Composition of Bone Morphogenetic Protein Human MP52 |
JP2000247903A (en) * | 1999-03-01 | 2000-09-12 | Chugai Pharmaceut Co Ltd | Long-term stabilized pharmaceutical preparation |
WO2000054797A2 (en) * | 1999-03-17 | 2000-09-21 | Novartis Ag | Pharmaceutical compositions comprising tgf-beta |
JP2003500456A (en) * | 1999-06-02 | 2003-01-07 | ヒューマン・ジェノム・サイエンシズ・インコーポレイテッド | Keratinocyte growth factor-2 preparation |
GB9930882D0 (en) * | 1999-12-30 | 2000-02-23 | Nps Allelix Corp | GLP-2 formulations |
ATE355849T1 (en) * | 2000-12-21 | 2007-03-15 | Nektar Therapeutics | STORAGE-Stable POWDER COMPOSITIONS WITH INTERLEUKIN-4 RECEPTOR |
US6887462B2 (en) * | 2001-04-09 | 2005-05-03 | Chiron Corporation | HSA-free formulations of interferon-beta |
AU2002366267B2 (en) * | 2001-11-19 | 2007-05-10 | Becton, Dickinson And Company | Pharmaceutical compositions in particulate form |
GB0207092D0 (en) * | 2002-03-26 | 2002-05-08 | Sod Conseils Rech Applic | Stable pharmaceutical composition containing factor VIII |
US7777006B2 (en) * | 2002-12-31 | 2010-08-17 | Csl Behring L.L.C. | Method for purification of alpha-1-antitrypsin |
-
2004
- 2004-11-10 EP EP04798463A patent/EP1685160A1/en not_active Withdrawn
- 2004-11-10 AU AU2004288854A patent/AU2004288854B2/en not_active Ceased
- 2004-11-10 JP JP2006538939A patent/JP2007534633A/en active Pending
- 2004-11-10 WO PCT/GB2004/004740 patent/WO2005047323A1/en active Application Filing
- 2004-11-10 CA CA002545458A patent/CA2545458A1/en not_active Abandoned
Non-Patent Citations (1)
Title |
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See references of WO2005047323A1 * |
Also Published As
Publication number | Publication date |
---|---|
CA2545458A1 (en) | 2005-05-26 |
JP2007534633A (en) | 2007-11-29 |
WO2005047323A1 (en) | 2005-05-26 |
AU2004288854B2 (en) | 2009-10-01 |
AU2004288854A1 (en) | 2005-05-26 |
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Legal Events
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PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
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RIN1 | Information on inventor provided before grant (corrected) |
Inventor name: GIBSON, HELEN Inventor name: BATHURST, IAN, C. Inventor name: PEMBERTON, PHILIP, A. Inventor name: NAYAR, RAJIV Inventor name: MANNING, MARK, G. Inventor name: BARR, PHILIP, J. |
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17Q | First examination report despatched |
Effective date: 20060831 |
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RIN1 | Information on inventor provided before grant (corrected) |
Inventor name: BATHURST, IAN, C. Inventor name: MANNING, MARK, G. Inventor name: BARR, PHILIP, J. Inventor name: PEMBERTON, PHILIP, A. Inventor name: GIBSON, HELEN Inventor name: NAYAR, RAJIV |
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DAX | Request for extension of the european patent (deleted) | ||
RIN1 | Information on inventor provided before grant (corrected) |
Inventor name: PEMBERTON, PHILIP, A. Inventor name: BARR, PHILIP, J. Inventor name: BATHURST, IAN, C. Inventor name: GIBSON, HELEN Inventor name: NAYAR, RAJIV Inventor name: MANNING, MARK, G. |
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RAP1 | Party data changed (applicant data changed or rights of an application transferred) |
Owner name: ARRIVA-PROMETIC INC. |
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RAP1 | Party data changed (applicant data changed or rights of an application transferred) |
Owner name: ARRIVA PHARMACEUTICALS, INC. |
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