DK3025728T3 - Fremgangsmåde til oprensning af prokaryotiske phenylalanin-ammoniak-lyasevarianter - Google Patents
Fremgangsmåde til oprensning af prokaryotiske phenylalanin-ammoniak-lyasevarianter Download PDFInfo
- Publication number
- DK3025728T3 DK3025728T3 DK15202466.7T DK15202466T DK3025728T3 DK 3025728 T3 DK3025728 T3 DK 3025728T3 DK 15202466 T DK15202466 T DK 15202466T DK 3025728 T3 DK3025728 T3 DK 3025728T3
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- Prior art keywords
- avpal
- variant
- pal
- pegylated
- phe
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/88—Lyases (4.)
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- 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
- A61K38/43—Enzymes; Proenzymes; Derivatives thereof
- A61K38/51—Lyases (4)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
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- 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
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y403/00—Carbon-nitrogen lyases (4.3)
- C12Y403/01—Ammonia-lyases (4.3.1)
- C12Y403/01024—Phenylalanine ammonia-lyase (4.3.1.24)
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Hematology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Immunology (AREA)
- Gastroenterology & Hepatology (AREA)
- Dermatology (AREA)
- Diabetes (AREA)
- Obesity (AREA)
- Oncology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicinal Preparation (AREA)
- Enzymes And Modification Thereof (AREA)
- Analytical Chemistry (AREA)
- Biophysics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Claims (13)
1. Fremgangsmåde til oprensning af en Anabaena variabilis-pbeny\a\an\n-ammoniak-lyase (AvPAL)-variant med minimal aggregering omfattende: (a) at lysere bakterieceller indeholdende AvPAL-varianten ved homogenisering for at generere et celle-lysat; (b) at opvarme celle-lysatet til 65 °C i 30 til 120 minutter; (c) at centrifugere det opvarmede celle-lysat, hvor en supernatant omfattende AvPAL-varianten tilbageholdes; (d) at filtrere supernatanten for at fjerne præcipitater; og (e) at separere AvPAL-varianten fra kontaminerende proteiner ved sekventiel kromatografi over en anionbytter-(AIEX)-søjle efterfulgt af en hydrofob interaktion-(HIC)-søjle, hvor eluatet fra HlC-søjlen omfatter AvPAL-varianten, hvor cystein-resterne i positionerne 503 og 565 af nævnte AvPAL-variant er blevet substitueret med serin-rester (SEQ ID NO: 11).
2. Fremgangsmåden ifølge krav 1, hvor fremgangsmåden er en fremgangsmåde til oprensning afen pegyleretAnabaena variabilis phenylalanin-ammoniak-lyase (AvPAL)-variant med minimal aggregering og hvor fremgangsmåden yderligere omfatter: (f) at ultrafiltrere eller ultrafiltrere/diafiltrere eluatet fra HlC-søjlen omfattende AvPAL-varianten; (g) at pegylere AvPAL-varianten ved at blande polyethylenglycol med AvPAL-varianten; (h) at fjerne fri polyethylenglycol fra den pegylerede AvPAL-variant ved ultrafiltation/diafiltration; og (i) at formulere den pegylerede AvPAL-variant, hvor nævnte AvPAL-variant omfatter polyethylenglycol.
3. Fremgangsmåden ifølge krav 2, hvor forholdet mellem AvPAL-variant og polyethylenglycolen er omkring 1:3.
4. Fremgangsmåden ifølge krav 2 eller 3, yderligere omfattende at fryse og optø eluatet fra HlC-søjlen omfattende AvPAL-varianten opnået i trin (e), hvor en eller flere polyoler eller sukre valgt fra gruppen bestående af glycerol, sukrose, glucose, trehalose, mannitol og sorbitol tilsættes til HIC-søjle-eluatet forud for frysning.
5. Fremgangsmåden ifølge krav 4, hvor polyolen er glycerol, fortrinsvis hvor koncentrationen af glycerol er 10 % (v/v).
6. Fremgangsmåden ifølge krav 4 eller 5, hvor sukkeret er sukrose, fortrinsvis hvor koncentrationen af sukrose er 10 % (v/v).
7. Fremgangsmåden ifølge et hvilket som helst af kravene 4 til 6, hvor at fryse eller optø eluatet fra HlC-søjlen omfattende AvPAL-varianten udføres i adskilte temperaturtrin.
8. Fremgangsmåden ifølge et hvilket som helst af kravene 4 til 7, hvor at fryse og optø eluatet fra HlC-søjlen omfattende AvPAL-varianten udføres i adskilte temperaturtrin.
9. Fremgangsmåden ifølge et hvilket som helst af kravene 2 til 8, hvor en diafiltreringsbuffer i trin (f) omfatter kaliumphosphat (KPi) og et eller flere stoffer bestående af trans-kanelsyre (t-CA) og glycerol, fortrinsvis hvor diafiltreringsbufferen omfatter 50 mM KPi, 10 mM t-CA, 5 % glycerol, pH 8,5.
10. Fremgangsmåden ifølge et hvilket som helst af kravene 2 til 9, hvor en ikke-ionisk detergent tilsættes til det ultrafiltrerede eller ultrafiltrerede/diafiltrerede eluat fra HlC-søjlen omfattende AvPAL-varianten opnået i trin (f).
11. Fremgangsmåden ifølge krav 10, hvor den ikke-ioniske detergent er polysorbat 80 (PS80), fortrinsvis hvor koncentrationen af PS80 er 0,02 %.
12. Fremgangsmåden ifølge et hvilket som helst af kravene 1 til 11, hvor AIEX-søjlen er en Toyopearl Giga Cap Q 650M-søjle.
13. Fremgangsmåden ifølge et hvilket som helst af kravene 1 til 12, hvor HIC-søjlen er en Toyopearl Butyl 650M-søjle.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US30147810P | 2010-02-04 | 2010-02-04 | |
| EP11740323.8A EP2531209B1 (en) | 2010-02-04 | 2011-02-03 | Compositions of prokaryotic phenylalanine ammonia-lyase variants and methods of using compositions thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DK3025728T3 true DK3025728T3 (da) | 2018-10-29 |
Family
ID=44356065
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK15202466.7T DK3025728T3 (da) | 2010-02-04 | 2011-02-03 | Fremgangsmåde til oprensning af prokaryotiske phenylalanin-ammoniak-lyasevarianter |
Country Status (18)
| Country | Link |
|---|---|
| US (4) | US20130039898A1 (da) |
| EP (4) | EP2531209B1 (da) |
| JP (1) | JP5828844B2 (da) |
| CN (2) | CN106834261B (da) |
| AU (1) | AU2011212929B2 (da) |
| CA (1) | CA2782444C (da) |
| CY (1) | CY1120768T1 (da) |
| DK (1) | DK3025728T3 (da) |
| ES (2) | ES2690542T3 (da) |
| HK (1) | HK1225299B (da) |
| HR (1) | HRP20181602T1 (da) |
| HU (1) | HUE039516T2 (da) |
| IL (2) | IL221099B (da) |
| LT (1) | LT3025728T (da) |
| PL (1) | PL3025728T3 (da) |
| PT (1) | PT3025728T (da) |
| SI (1) | SI3025728T1 (da) |
| WO (1) | WO2011097335A2 (da) |
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| ES2690542T3 (es) | 2010-02-04 | 2018-11-21 | Biomarin Pharmaceutical Inc. | Método para purificar variantes de fenilalanina amoníaco-liasa procariotas |
| NZ713396A (en) | 2013-04-18 | 2020-06-26 | Codexis Inc | Engineered phenylalanine ammonia lyase polypeptides |
| WO2015161019A1 (en) | 2014-04-16 | 2015-10-22 | Codexis, Inc. | Engineered tyrosine ammonia lyase |
| TWI743023B (zh) * | 2014-05-21 | 2021-10-21 | 日商Ｋｍ生物醫藥股份有限公司 | 基質金屬蛋白酶 7(mmp-7)聚集體之單體化方法 |
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| BR112022018571A2 (pt) * | 2020-03-20 | 2022-11-01 | Synlogic Operating Co Inc | Microrganismos manipulados para reduzir a hiperfenilalaninemia |
| CN116897205A (zh) | 2020-08-28 | 2023-10-17 | 科德克希思公司 | 工程化淀粉酶变体 |
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| EP4341394A4 (en) | 2021-05-19 | 2025-04-09 | BioMarin Pharmaceutical Inc. | Compositions of prokaryotic phenylalanine ammonia lyase and methods for treating adolescent patients |
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| WO2010014225A2 (en) | 2008-07-30 | 2010-02-04 | Biomarin Pharmaceutical Inc. | Assays for detection of phenylalanine ammonia-lyase and antibodies to phenylalanine ammonia-lyase |
| ES2690542T3 (es) | 2010-02-04 | 2018-11-21 | Biomarin Pharmaceutical Inc. | Método para purificar variantes de fenilalanina amoníaco-liasa procariotas |
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