CN104826117B - Storage stabilizing agent for human serum immunoglobulin solution preparation - Google Patents
Storage stabilizing agent for human serum immunoglobulin solution preparation Download PDFInfo
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- CN104826117B CN104826117B CN201510221517.8A CN201510221517A CN104826117B CN 104826117 B CN104826117 B CN 104826117B CN 201510221517 A CN201510221517 A CN 201510221517A CN 104826117 B CN104826117 B CN 104826117B
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Abstract
The present invention discloses a kind of stabilizer that human serum immunoglobulin solution preparation is reduced or avoided and polymerize in storage process; containing L cysteines or its hydrochloride, one or more of mercapto-protective agents of glutathione, its concentration it is 3-20mmol/L in immunoglobulin solution preparation；One or more of sugared stabilizers containing maltose or trehalose, its concentration are 150-600mmol/L；And one or more of amino acid stabilizers containing proline or its salt, glycine or its salt, serine or its salt, its concentration is 12-150mmol/L, by adding stabilizer, immunoglobulin can be reduced because of high temperature, high concentration or storage time is longer and the polymerization that occurs, improve the validity of medicine.
Description
Technical field
The present invention relates to medicated premix, the storage of human serum immunoglobulin solution preparation specifically used for intravenous injection
Deposit the stabilizer used.
Background technology
Human serum immunoglobulin（IgG）Preparation is mainly used in primary and Secondary cases immunoglobulin deficiency, with
And other autoimmune diseases, such as primary thrombocytopenic purpura, Kawasaki disease.
IgG is to purify to obtain from human plasma, and it after B cell and thick liquid cell contact antigen by producing, and its molecular weight is about
For 150KDa, it is made up of four peptide chains, two identical light chains（L）And heavy chain（H）It is covalently attached to by two disulfide bond,
In addition, a Y type dimer symmetrically is formed with non-covalent.Light chain is made up of two homeodomains, weight
Chain is made up of four homeodomains, and these domains are characterized in including 2 beta sheet structures, including 3 to 5 anti-phase parallel
Beta chain, Stability Analysis of Structures is maintained by a disulfide bond.These domains are to thermo-responsive, and its secondary structure can under room temperature storage
Change, result in polymer, immunoglobulin preparation storage process polymer is continuously increased, and drug effect gradually reduces, and leads to
After crossing intravenous injection, polymer can also produce side effect.
CN103282042A discloses a kind of with histidine and the stable storing under faintly acid to neutral pH immune ball
Albumen aqueous formulation, including immunoglobulin, the histidine from 50mM to 500mM, the alkali metal cation from 0mM to 10mM,
And from 5.5 to 7.0 pH, immunoglobulin can be made stable under faintly acid to neutrallty condition.But high temperature, high concentration or
When storage time is longer, immunoglobulin extent of polymerization is still higher.
The content of the invention
Human serum immunoglobulin solution preparation can be reduced or avoided it is an object of the invention to provide one kind to store
The stabilizer of Cheng Fasheng polymerizations, by adding stabilizer, can reduce immunoglobulin because high temperature, high concentration or storage time are longer
And the polymerization occurred, improve the validity of medicine.
The storage stabilizing agent of the present invention is to contain Cys or its hydrochloride, paddy Guang in immunoglobulin solution preparation
One or more of mercapto-protective agents of sweet peptide, its concentration are 3-20mmol/L；One kind containing maltose or trehalose or with
On sugared stabilizer, its concentration is 150-600mmol/L；And contain proline or its salt, glycine or its salt, serine
Or one or more of amino acid stabilizers of its salt, its concentration are 12-150mmol/L.
Preferably, the molar concentration of mercapto-protective agent is 4-8mmol/L, the molar concentration of sugared stabilizer for 250-
400mmol/L, the molar concentration of amino acid stabilizers is 20-80mmol/L.
The pH value of immunoglobulin solution preparation is preferably 4-6.
Mercapto-protective agent used herein can protect immune globulin intramolecular disulfide bond, maintain its three-level and level Four knot
Structure, sugared stabilizer are used for the secondary structure stabilization for protecting β-pleated sheet in immunoglobulin molecules, and amino acid stabilizers can promote to be immunized
Uniform separation between globulin molecule, stablizes its native conformation.Thus immunoglobulin solution preparation can be avoided or reduce in length
Phase storage process further polymerize, and improves storage stability, and formulation storage is immunized after 2 years in room temperature and the following environment of room temperature
Globulin poly aggressiveness is less than 1%, and for formulation storage after 2 years, immunoglobulin polymer is less than 2% in 37 DEG C of environment.
Embodiment
Embodiment 1
Learn from else's experience the human serum chromatography of immunoglobulins solution after inactivation of virus（Purity 99.2%, concentration 0.37mmol/L）,
The immunoglobulin solution for being configured to that concentration is 10wt% is concentrated by ultrafiltration, takes 1L samples, addition glucose to 230mmol/L, regulation
PH to 4.1 is used as blank control sample, separately takes 3L, addition glutathione to 6mmol/L, maltose to 300mmol/L, proline
To 50mmol/L, pH to 4.1 is adjusted, is divided into sample 1, sample 2,3 each 1L of sample.Product is big to molecule under the conditions of room temperature storage
Small distribution is investigated.
Molecular size distribution is checked using high performance liquid chromatography：With hydrophilic silica gels Efficient numerical method post
（SEC, exclusion limit 300kD, 10 μm of granularity）, column diameter 7.5mm, long 60cm；With containing 1% isopropanol, pH value 7.0,0.2mol/L
Phosphate buffer（Take 0.5mol/L sodium dihydrogen phosphate 200ml, 0.5mol/L disodium hydrogen phosphates 420ml, isopropanol 15.5ml and
Water 914.5ml is uniformly mixed）For mobile phase；Detection wavelength is 280nm；Flow velocity is 0.6ml per minute；Every 1ml is taken to contain egg respectively
White matter is 12mg each 20 μ l of immunoglobulin, human serum albumin solution, is injected separately into chromatographic column, records chromatogram, and ball is immunized
The separation at protein control monomer peak and cracking body peak should be greater than 1.5, point of human serum albumin reference substance monomer peak and dimer peak
1.5 are should be greater than from degree, tailing factor is calculated by human serum albumin monomer peak and should be 0.95-1.40.Calculated by area normalization method,
Monomer adds the content at dimer peak in chromatogram, and as immunoglobulin monomer adds dimer content.
Measure immunoglobulin monomer adds dimer content when storing 3,6,9,12 and 24 months, as a result such as following table.
Embodiment 2
Learn from else's experience the human serum chromatography of immunoglobulins solution after inactivation of virus（Purity 99.5%, concentration 0.25mmol/L）,
The immunoglobulin solution for being configured to that concentration is 10wt% is concentrated by ultrafiltration, takes 1L samples, addition glucose to 220mmol/L, regulation
PH to 3.85 is used as blank control sample, separately takes 3L, addition cysteine to 4mmol/L, maltose to 500mmol/L, sweet ammonia
Acid adjusts pH to 3.85, respectively sample 1, sample 2, sample 3 to 130mmol/L.Product is under 37 DEG C of conditions of storage to molecule
Size distribution is investigated.
Molecular size is distributed according to embodiment 1, is checked using high performance liquid chromatography.Storage 3,6,9,12 and 24
Measure immunoglobulin monomer adds dimer content during the moon, as a result such as following table.
Embodiment 3
Learn from else's experience the chromatography of immunoglobulins solution after inactivation of virus（Purity 98.7%, concentration 0.27mmol/L）, it is concentrated by ultrafiltration
The immunoglobulin solution that concentration is 5wt% is configured to, takes 1L samples, addition glucose to 225mmol/L, regulation pH to 6.9 makees
For blank control sample, 3L, addition glutathione to 8mmol/L, maltose to 250mmol/L, serine to 75mmol/ are separately taken
L, pH to 6.9 is adjusted, is divided into sample 1, sample 2,3 each 1L of sample.Product molecular size is distributed under 37 DEG C of conditions of storage into
Row is investigated.
Molecular size is distributed according to embodiment 1, is checked using high performance liquid chromatography.Storage 3,6,9,12 and 24
Measure immunoglobulin monomer adds dimer content during the moon, as a result such as following table.
Embodiment 4
Learn from else's experience the human serum chromatography of immunoglobulins solution after inactivation of virus（Purity 99.0%, concentration 0.26mmol/L）,
The immunoglobulin solution for being configured to that concentration is 5wt% is concentrated by ultrafiltration.1L samples are taken, add glucose 200mmol/L, adjust pH
Blank control sample is used as to 5.5.Another to take 3L, addition glutathione is to 8mmol/L, and maltose to 350mmol/L, glycine is extremely
80mmol/L, pH to 5.5 is adjusted, is divided into sample 1, sample 2,3 each 1L of sample.Product is under 15 DEG C of conditions of storage to molecular size
Distribution is investigated.
Molecular size is distributed according to embodiment 1, is checked using high performance liquid chromatography.Storage 3,6,9,12 and 24
Measure immunoglobulin monomer adds dimer content during the moon, as a result such as following table.
Claims (4)
- A kind of 1. storage stabilizing agent for human serum immunoglobulin solution preparation, it is characterised in that：The immune globulin Contain Cys or its hydrochloride, one or more of mercapto-protective agents of glutathione, its concentration in white pharmaceutical solutions For 3-20mmol/L；One or more of sugared stabilizers containing maltose or trehalose, its concentration are 150-600mmol/ L；And one or more of amino acid stabilizers containing proline or its salt, glycine or its salt, serine or its salt, its Concentration is 12-150mmol/L, immunoglobulin solution preparation pH=4-6.
- 2. storage stabilizing agent according to claim 1, it is characterised in that the concentration of the mercapto-protective agent is 4-10mmol/ L。
- 3. storage stabilizing agent according to claim 1, it is characterised in that the concentration of the sugared stabilizer be 250- 400mmol/L。
- 4. storage stabilizing agent according to claim 1, it is characterised in that the concentration of the amino acid stabilizers be 20- 80mmol/L。
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| CN104826117B true CN104826117B (en) | 2017-11-14 |
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| US6290967B1 (en) * | 1996-12-20 | 2001-09-18 | Merck & Co., Inc. | Stabilizers for lyophilized vaccines |
| KR101212025B1 (en) * | 2002-06-21 | 2013-01-09 | 노보 노르디스크 헬스 케어 악티엔게젤샤프트 | Stabilised solid compositions of factor ⅶ polypeptides |
| TW201213342A (en) * | 2010-09-17 | 2012-04-01 | Baxter Int | Stabilization of immunoglobulins and other proteins through aqueous formulation with sodium chloride at weak acidic to neutral pH |
| CN103550780B (en) * | 2013-10-30 | 2015-03-18 | 郑州邦和生物药业有限公司 | Protein protective agent for Pasteur inactivating human intravenous immunoglobulin and inactivation method of protein protective agent |
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