HRP20240797T3 - Pripravak topljive hijaluronidaze - Google Patents
Pripravak topljive hijaluronidazeInfo
- Publication number
- HRP20240797T3 HRP20240797T3 HRP20240797TT HRP20240797T HRP20240797T3 HR P20240797 T3 HRP20240797 T3 HR P20240797T3 HR P20240797T T HRP20240797T T HR P20240797TT HR P20240797 T HRP20240797 T HR P20240797T HR P20240797 T3 HRP20240797 T3 HR P20240797T3
- Authority
- HR
- Croatia
- Prior art keywords
- volume
- nutrient medium
- culture
- alanyl
- glutamine
- Prior art date
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Classifications
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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
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2474—Hyaluronoglucosaminidase (3.2.1.35), i.e. hyaluronidase
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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
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01035—Hyaluronoglucosaminidase (3.2.1.35), i.e. hyaluronidase
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01036—Hyaluronoglucuronidase (3.2.1.36)
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Claims (16)
1. Pripravak koji sadrži mješavinu polipeptida, gdje navedena mješavina sadrži polipeptide prikazane u SEQ ID NOS: 4-8, naznačen time što se pripravak može dobiti sljedećim postupkom:
a) inokulacijom staničnog medija u bioreaktoru sa inokulumom stanica koje kodiraju topljivi rekombinantni ljudski PH20 (rHuPH20) za proizvodnju stanične kulture, pri čemu:
stanice sadrže između 150 i 300 kopija nukleinske kiseline koja kodira topljivi rHuPH20;
bioreaktor sadrži najmanje 100 litara stanične kulture;
1010 - 1011 stanica je inokulirano na 100 litara stanične kulture; i
stanice se kultiviraju na podešenoj temperaturi;
b) hranjenje stanica prvim hranjivim medijem koji sadrži glukozu, L-alanil-L-glutamin, ljudski inzulin i ekstrakt kvasca u količinama dovoljnim da povećaju rast stanica i pik stanične gustoće, i da povećaju sintezu topljivog rHuPH20, pri čemu se hranjivi medij dodaje kulturi u zapremini od 0.5% do 20% zapremine stanične kulture;
c) hranjenje stanica drugim hranjivim medijem koji sadrži glukozu, L-alanil-L-glutamin, ekstrakt kvasca i natrij butirat u količinama dovoljnim da povećaju sintezu topljivog rHuPH20 i induciraju zaustavljanje staničnog ciklusa; i snižavanje temperature u usporedbi sa temperaturom u koraku a) na temperaturu dovoljnu da poveća zaustavljanje staničnog ciklusa, poveća vitalnost stanice i stabilizira topljivu hijaluronidazu; pri čemu:
količina L-alanil-L-glutamina je smanjena u usporedbi sa količinom L-alanil-L-glutamina u koraku b);
količina ekstrakta kvasca je povećana u usporedbi sa količinom ekstrakta kvasca u koraku b); i
hranjivi medij se dodaje kulturi u zapremini od 0.5% do 20% zapremine stanične kulture;
d) hranjenje stanica trećim hranjivim medijem koji sadrži glukozu, L-alanil-L-glutamin, ekstrakt kvasca i natrij butirat u količinama dovoljnim da povećaju sintezu topljivog rHuPH20 i povećaju zaustavljanje staničnog ciklusa, i snižavanje temperature u usporedbi sa temperaturom u koraku c) do temperature dovoljne da poveća zaustavljanje staničnog ciklusa, poveća vitalnost stanice i stabilizira topljivu hijaluronidazu; pri čemu:
količina L-alanil-L-glutamina je smanjena u usporedbi sa količinom L-alanil-L-glutamina u koraku c);
količine ekstrakta kvasca, glukoze i natrij butirata su povećane u usporedbi sa količinama ekstrakta kvasca, glukoze i natrij butirata u koraku c); i
hranjivi medij se dodaje kulturi u zapremini od 0.5% do 20% zapremine stanične kulture;
e) hranjenje stanica četvrtim hranjivim medijem koji sadrži glukozu, L-alanil-L-glutamin, ekstrakt kvasca i natrij butirat u količinama dovoljnim da povećaju sintezu topljivog rHuPH20 i povećaju zaustavljanje staničnog ciklusa, i snižavanje temperature u usporedbi sa temperaturom u koraku d) do temperature dovoljne da poveća zaustavljanje staničnog ciklusa, poveća vitalnost stanice i stabilizira topljivu hijaluronidazu; pri čemu:
količina L-alanil-L-glutamina i glukoze je smanjena u usporedbi sa količinom L-alanil-L-glutamina i glukoze u koraku d);
količina natrij butirata je smanjena u usporedbi sa količinom natrij butirata u koraku d); i
hranjivi medij se dodaje kulturi u zapremini od 0.5% do 20% zapremine stanične kulture;
f) nastavak kultiviranja stanica sve dok vitalnost ne padne ispod najmanje 50%;
g) dobivanje tekućine sakupljene stanične kulture; i
h) pročišćavanje rHuPH20 iz tekućine sakupljene stanične kulture,
pri čemu je topljivi rHuPH20 topljivi oblik ljudskog PH20 koji je rekombinantno eksprimiran u stanicama jajnika kineskog hrčka (CHO).
2. Pripravak prema patentnom zahtjevu 1, naznačen time što je temperatura u koraku a) 37 °C.
3. Pripravak prema patentnom zahtjevu 1 ili 2, naznačen time što je temperatura u koraku c) 36.5 °C.
4. Pripravak prema bilo kojem patentnom zahtjevu 1-3, naznačen time što je temperatura u koraku d) 36 °C.
5. Pripravak prema bilo kojem patentnom zahtjevu 1-4, naznačen time što je temperatura u koraku e) 35.5 °C.
6. Pripravak prema bilo kojem patentnom zahtjevu 1-5, naznačen time što se tekućina sakupljene stanične kulture filtrira prije pročišćavanja.
7. Pripravak prema bilo kojem patentnom zahtjevu 1-6, naznačen time što se pročišćavanje topljivog rHuPH20 vrši kromatografijom na koloni.
8. Pripravak prema patentnom zahtjevu 7, naznačen time što kromatografija na koloni obuhvaća kromatografiju na koloni sa agaroznim kuglicama, kromatografiju na koloni sa fenil supstituiranim agaroznim kuglicama, kromatografiju na koloni sa amino fenil boronatom i kromatografiju na koloni sa hidroksiapatitom.
9. Pripravak prema bilo kojem patentnom zahtjevu 1-8, naznačen time što se hranjivi medij dodaje u kulturu u zapremini od 4% zapremine stanične kulture.
10. Pripravak prema patentnom zahtjevu 1, naznačen time što:
u koraku a) stanice se kultiviraju na 37 °C;
u koraku b) prvi hranjivi medij sadrži 33 g/L glukoze, 32 mM L-alanil-L-glutamina, 16.6 g/L ekstrakta kvasca i 33 mg/L inzulina, i hranjivi medij se dodaje kulturi u zapremini od 4% zapremine stanične kulture;
u koraku c) drugi hranjivi medij sadrži 33 g/L glukoze, 16 mM L-alanil-L-glutamina, 33.4 g/L ekstrakta kvasca i 0.92 g/L natrij butirata, i hranjivi medij se dodaje kulturi u zapremini od 4% zapremine stanične kulture; i
temperatura se spušta na 36.5 °C;
u koraku d) treći hranjivi medij sadrži 50 g/L glukoze, 10 mM L-alanil-L-glutamina, 50 g/L ekstrakta kvasca i 1.8 g/L natrij butirata, i hranjivi medij se dodaje kulturi u zapremina od 4% zapremine stanične kulture; i
temperatura se spušta na 36 °C;
u koraku e) četvrti hranjivi medij sadrži 33 g/L glukoze, 6.6 mM L-alanil-L-glutamina, 50 g/L ekstrakta kvasca i 0.92 g/L natrij butirata, i hranjivi medij se dodaje u kulturu u zapremini od 4% zapremine stanične kulture; i
temperatura se spušta na 35.5 °C; dalje sadrži filtriranje tekućine sakupljene stanične kulture dobivene u koraku g) i
u koraku h) pročišćavanje rHuPH20 iz tekućine sakupljene kulture korištenjem kromatografije na koloni sa agaroznim kuglicama, kromatografije na koloni sa fenil supstituiranim agaroznim kuglicama, kromatografije na koloni sa amino fenil boronatom ili kromatografije na koloni sa hidroksiapatitom.
11. Pripravak prema patentnom zahtjevu 1, naznačen time što:
u koraku a) stanice se inokuliraju na gustoći stanica od 4 × 105 stanica/mL; i
stanice se kultiviraju na 37 °C;
u koraku b) prvi hranjivi medij sadrži 33 g/L glukoze, 32 mM L-alanil-L-glutamina, 83.3 g/L ekstrakta kvasca i 33 mg/L inzulina, i hranjivi medij se dodaje kulturi u zapremini od 4% zapremine stanične kulture;
u koraku c) drugi hranjivi medij sadrži 33 g/L glukoze, 13 mM L-alanil-L-glutamina, 166.7 g/L ekstrakta kvasca i 0.92 g/L natrij butirata, i hranjivi medij se dodaje u kulturu u zapremini od 4% zapremine stanične kulture; i
temperatura se spušta na 36.5 °C;
u koraku d) treći hranjivi medij sadrži 50 g/L glukoze, 10 mM L-alanil-L-glutamina, 250 g/L ekstrakta kvasca i 1.8 g/L natrij butirata, i hranjivi medij se dodaje kulturi u zapremini od 4% zapremine stanične kulture; i
temperatura se spušta na 36 °C;
u koraku e) četvrti hranjivi medij sadrži 33 g/L glukoze, 6.7 mM L-alanil-L-glutamina, 250 g/L ekstrakta kvasca i 0.92 g/L natrij butirata, i hranjivi medij se dodaje kulturi u zapremini od 4% zapremine stanične kulture; i
temperatura se spušta na 35.5 °C; dalje sadrži
filtriranje tekućine sakupljene stanične kulture dobivene u koraku g) i
u koraku h) pročišćavanje rHuPH20 iz tekućine sakupljene kulture korištenjem kromatografije na koloni sa agaroznim kuglicama, kromatografije na koloni sa fenil supstituiranim agaroznim kuglicama, kromatografije na koloni sa amino fenil boronatom ili kromatografije na koloni sa hidroksiapatitom.
12. Pripravak prema bilo kojem patentnom zahtjevu 1-11, naznačen time što se najmanje 0.5, 1, 5, 10, 15, 20, 25, 30, 35 ili 40 grama topljivog rHuPH20 proizvodi na 100 L stanične kulture.
13. Pripravak prema bilo kojem patentnom zahtjevu 1-12, naznačen time što je specifična aktivnost topljivog rHuPH20 najmanje 80000, 100000, 120000, 140000, 160000 ili 180 000 jedinica/mg.
14. Pripravak prema bilo kojem patentnom zahtjevu 1-13, naznačen time što je zapremina stanične kulture u bioreaktoru 200, 300, 400, 500, 1000, 1500, 2000, 2500, 3000 ili 3500 litara.
15. Pripravak prema bilo kojem patentnom zahtjevu 1-14, naznačen time što su stanice koje kodiraju topljivi rHuPH20 stanice DG44 CHO.
16. Pripravak prema bilo kojem patentnom zahtjevu 1-15, naznačen time što je rHuPH20 kodiran nukleinskom kiselinom prikazanom u SEQ ID NO:47.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US6862208P | 2008-03-06 | 2008-03-06 | |
| EP23191930.9A EP4269578B8 (en) | 2008-03-06 | 2009-03-06 | Soluble hyaluronidase composition |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| HRP20240797T3 true HRP20240797T3 (hr) | 2024-09-13 |
Family
ID=40732065
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| HRP20240797TT HRP20240797T3 (hr) | 2008-03-06 | 2009-03-06 | Pripravak topljive hijaluronidaze |
Country Status (18)
| Country | Link |
|---|---|
| US (2) | US8187855B2 (hr) |
| EP (6) | EP2674487A3 (hr) |
| JP (2) | JP5650546B2 (hr) |
| KR (2) | KR20140130512A (hr) |
| CN (2) | CN106906196A (hr) |
| CA (1) | CA2717383C (hr) |
| DK (1) | DK4269578T3 (hr) |
| ES (3) | ES2981983T3 (hr) |
| FI (1) | FI4269578T3 (hr) |
| HR (1) | HRP20240797T3 (hr) |
| HU (1) | HUE067606T2 (hr) |
| IL (2) | IL207398A (hr) |
| LT (1) | LT4269578T (hr) |
| PL (1) | PL4269578T3 (hr) |
| PT (1) | PT4269578T (hr) |
| RS (1) | RS65606B1 (hr) |
| SI (1) | SI4269578T1 (hr) |
| WO (1) | WO2009111066A1 (hr) |
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