DK2970512T3 - Immunmodulatoriske fusionsproteiner og fremgangsmåder til fremstilling deraf - Google Patents
Immunmodulatoriske fusionsproteiner og fremgangsmåder til fremstilling deraf Download PDFInfo
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- DK2970512T3 DK2970512T3 DK14779937.3T DK14779937T DK2970512T3 DK 2970512 T3 DK2970512 T3 DK 2970512T3 DK 14779937 T DK14779937 T DK 14779937T DK 2970512 T3 DK2970512 T3 DK 2970512T3
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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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/71—Receptors; Cell surface antigens; Cell surface determinants for growth factors; for growth regulators
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
- C07K16/2818—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD28 or CD152
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2863—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators
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- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/62—DNA sequences coding for fusion proteins
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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
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/40—Immunoglobulins specific features characterized by post-translational modification
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/52—Constant or Fc region; Isotype
- C07K2317/526—CH3 domain
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
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- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
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- Proteomics, Peptides & Aminoacids (AREA)
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- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
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- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
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Claims (16)
1. Fremgangsmåde til fremstilling af et terapeutisk aktivt fusionsprotein, hvor fusionsproteinet omfatter en tumor-targeting del og mindst et immunmodulato-risk molekyle, der modvirker en cancercelles immunologiske tolerance, hvor den tumor-targeting del er et antistof, der binder til EGFR1 eller CTLA-4, og hvor fusionsproteinet fremstilles ved følgende trin: at fremstille en kodon-optimeret nukleotidsekvens, der koder for fusionsproteinet til ekspression i CHO-celler, hvor den kodon-optimerede nukleotidsekvens er modificeret til øgning af CG-sekvenser, og hvor den kodon-optimerede nukleotidsekvens er SEQ ID NO: 1 eller 5 for den tunge kæde af antistoffet, SEQ ID NO: 2 eller 6 for den lette kæde af antistoffet, SEQ ID NO: 3 eller 7 for det immunmodulatoriske molekyle, hvor den kodon-optimerede nukleotidsekvens SEQ ID NO: 1 eller 5 for den tunge kæde af antistoffet, der binder til henholdsvis EGFR1 eller CTLA-4, ikke har nukleotider til ekspression af et lysin ved den C-terminale ende af den tunge kæde af antistoffet; at klone den optimerede sekvens af fusionsproteinet i de CHO-celler, der er i stand til transient eller stabil ekspression; at dyrke CHO-cellerne i et medium under betingelser, der er egnede til dyrkning, og lade værtscellen eksprimere fusionsproteinet; og at indsamle secernerede fusionsproteiner og eventuelt til yderligere oprensning.
2. Fremgangsmåde ifølge krav 1, hvor det immunmodulatoriske molekyle er bundet til antistoffet gennem en aminosyresekvens eksprimeret ved SEQ ID NO: 4 af tilstrækkelig længde til at muliggøre bispecifik binding af fusionsproteinet og er bundet direkte eller gennem en linker til den tunge kæde af antistoffet, den lette kæde af antistoffet eller begge kæder.
3. Fremgangsmåde ifølge krav 1, hvor det immunmodulatoriske molekyle er bundet direkte eller gennem en linker eksprimeret ved SEQ ID NO: 4 til N- eller C-terminus af den tunge kæde af antistoffet, N- eller C-terminus af den lette kæde af antistoffet eller både N- og C-terminus af begge kæder.
4. Fremgangsmåde ifølge krav 1, hvor de kodon-optimerede nukleotidsekven-ser omfatter SEQ ID NOs: 1,2, 4 og 7.
5. Fremgangsmåde ifølge krav 1, hvor de kodon-optimerede nukleotidsekven-ser omfatter SEQ ID NOs: 1,2, 3 og 4.
6. Fremgangsmåde ifølge krav 1, hvor de kodon-optimerede nukleotidsekven-ser omfatter SEQ ID NOs: 5, 6, 7 og 4.
7. Præparat omfattende homogene terapeutisk aktive fusionsproteiner, hvor fusionsproteinerne omfatter en tumor-targeting del og mindst et immunmodu-latorisk molekyle, hvor den tumor-targeting del er et antistof, der binder til EGFR1 eller CTLA-4, og hvor fusionsproteinerne er fremstillet i henhold til krav 1, hvor fusionsproteinet har en kodon-optimeret nukleotidsekvens, der koder for fusionsproteinet til ekspression i CHO-celler, hvor den kodon-optimerede nukleotidsekvens er modificeret til øgning af CG-sekvenser, og hvor den kodon-optimerede nukleotidsekvens er SEQ ID NO: 1 eller 5 for den tunge kæde af antistoffet, SEQ ID NO: 2 eller 6 for den lette kæde af antistoffet, SEQ ID NO: 3 eller 7 for det immunmodulatoriske molekyle, hvor den kodon-optimerede nukleotidsekvens SEQ ID NO: 1 eller 5 for den tunge kæde af antistoffet, der binder til henholdsvis EGFR1 eller CTLA-4, ikke har nukleotider til ekspression af et lysin ved den C-terminale ende af den tunge kæde af antistoffet.
8. Nukleinsyresekvens, der koder for det kimære fusionsprotein ifølge krav 1, hvor den tunge kæde og lette kæde er SEQ ID NO: 1 og 2 eller SEQ ID NO: 5 og 6, hvor den immunmodulerende target-del er SEQ ID NO: 3 eller 7.
9. Vektor omfattende nukleinsyresekvenserne ifølge krav 8.
10. Fremgangsmåde ifølge krav 1, hvor fusionsproteinet omfatter aminosyre-sekvenserne med SEQ ID NOs: 9 og en blandt SEQ ID NOs: 15, 17, 27 eller 29.
11. Fremgangsmåde ifølge krav 1, hvor fusionsproteinet omfatter aminosyre-sekvenserne med SEQ ID NOs: 8 og en blandt SEQ ID NOs: 16, 18, 28 eller 30.
12. Fremgangsmåde ifølge krav 1, hvor fusionsproteinet omfatter aminosyre-sekvenserne med SEQ ID NOs: 31 og en blandt SEQ ID NOs: 28 eller 30.
13. Fremgangsmåde ifølge krav 1, hvor fusionsproteinet omfatter aminosyre-sekvenserne med SEQ ID NOs: 29 og en blandt SEQ ID NOs: 28 eller 30.
14. Fremgangsmåde ifølge krav 1, hvor fusionsproteinet omfatter aminosyre-sekvenserne med SEQ ID NOs: 32 og en blandt SEQ ID NOs: 14, 33 eller 35.
15. Fremgangsmåde ifølge krav 1, hvor fusionsproteinet omfatter aminosyre-sekvenserne med SEQ ID NOs: 13 og en blandt SEQ ID NOs: 33 eller 35.
16. Fremgangsmåde ifølge krav 1, hvor fusionsproteinet omfatter aminosyre-sekvenserne med SEQ ID NOs: 34 og en blandt SEQ ID NOs: 14, 33 eller 35.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201361777016P | 2013-03-12 | 2013-03-12 | |
| PCT/US2014/022404 WO2014164427A1 (en) | 2013-03-12 | 2014-03-10 | Fusion immunomodulatory proteins and methods for making same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DK2970512T3 true DK2970512T3 (da) | 2019-01-14 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK14779937.3T DK2970512T3 (da) | 2013-03-12 | 2014-03-10 | Immunmodulatoriske fusionsproteiner og fremgangsmåder til fremstilling deraf |
Country Status (16)
| Country | Link |
|---|---|
| US (3) | US9988456B2 (da) |
| EP (1) | EP2970512B1 (da) |
| JP (2) | JP6605444B2 (da) |
| CN (1) | CN105121474B9 (da) |
| AU (2) | AU2014249405C1 (da) |
| BR (1) | BR112015022733B1 (da) |
| CA (2) | CA2902830C (da) |
| DK (1) | DK2970512T3 (da) |
| ES (1) | ES2704411T3 (da) |
| HK (1) | HK1217343A1 (da) |
| MY (1) | MY186864A (da) |
| NZ (1) | NZ711445A (da) |
| PL (1) | PL2970512T3 (da) |
| PT (1) | PT2970512T (da) |
| RU (2) | RU2662991C2 (da) |
| WO (1) | WO2014164427A1 (da) |
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| PT3489254T (pt) * | 2012-04-30 | 2022-12-30 | Biocon Ltd | Proteínas de fusão direcionadas/imunomoduladoras e seus métodos de fabrico |
| RU2662991C2 (ru) * | 2013-03-12 | 2018-07-31 | Биокон Лтд. | Слитые иммуномодулирующие белки и способы их получения |
| JP2018167875A (ja) * | 2017-03-30 | 2018-11-01 | 株式会社フジシール | 容器包装体及び容器包装体の製造方法 |
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