HRP20171777T1 - Novi himerni polipeptidi za potrebe probira i otkrivanja lijekova - Google Patents

Novi himerni polipeptidi za potrebe probira i otkrivanja lijekova Download PDF

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Publication number
HRP20171777T1
HRP20171777T1 HRP20171777TT HRP20171777T HRP20171777T1 HR P20171777 T1 HRP20171777 T1 HR P20171777T1 HR P20171777T T HRP20171777T T HR P20171777TT HR P20171777 T HRP20171777 T HR P20171777T HR P20171777 T1 HRP20171777 T1 HR P20171777T1
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Croatia
Prior art keywords
gpcr
chimeric polypeptide
conformation
polypeptide according
cell
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HRP20171777TT
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English (en)
Inventor
Jan Steyaert
Toon Laeremans
Els Pardon
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Vib Vzw
Vrije Universiteit Brussel
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Publication of HRP20171777T1 publication Critical patent/HRP20171777T1/hr

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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
    • 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
    • C07K14/72—Receptors; Cell surface antigens; Cell surface determinants for hormones
    • C07K14/723—G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH receptor
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/26—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against hormones ; against hormone releasing or inhibiting factors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/286—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against neuromediator receptors, e.g. serotonin receptor, dopamine receptor
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2869—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against hormone receptors
    • 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
    • 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/566—Immunoassay; Biospecific binding assay; Materials therefor using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/22—Immunoglobulins specific features characterized by taxonomic origin from camelids, e.g. camel, llama or dromedary
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/32—Immunoglobulins specific features characterized by aspects of specificity or valency specific for a neo-epitope on a complex, e.g. antibody-antigen or ligand-receptor
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/569—Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/75—Agonist effect on antigen
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/74—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/72—Assays involving receptors, cell surface antigens or cell surface determinants for hormones
    • G01N2333/726—G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2500/00—Screening for compounds of potential therapeutic value
    • G01N2500/04—Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Endocrinology (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • General Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Neurology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Toxicology (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Food Science & Technology (AREA)
  • Plant Pathology (AREA)
  • Peptides Or Proteins (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Claims (15)

1. Himerni polipeptid koji sadržava GPCR fuzioniran na konformacijsku selektivnu veznu domenu, naznačen time, da je navedena vezna domena pojedinačna varijabilna domena imunoglobulina i pri čemu je navedena vezna domena usmjerena na i / ili sposobna specifično vezati GPCR, pri čemu je GPCR stabiliziran u aktivnoj ili neaktivnoj konformaciji nakon vezanja vezne domene i pri čemu navedeni GPCR, koji je stabiliziran u navedenoj aktivnoj ili neaktivnoj konformaciji, ima povećani afinitet za konformacijski selektivni ligand u usporedbi s odgovarajućim nefuzioniranim GPCR-om.
2. Himerni polipeptid u skladu s patentnim zahtjevom 1, naznačen time, da je domena vezanja spojena s GPCR-om, bilo izravno ili putem povezivača.
3. Himerni polipeptid u skladu s patentnim zahtjevom 1 do 2, naznačen time, da je aktivna konformacija konformacija agonista, konformacija djelomičnog agonista ili konformacija agonista s odstupanjem.
4. Himerni polipeptid u skladu s patentnim zahtjevom 1 do 2, naznačen time, da je neaktivna konformacija inverzna konformacija agonista.
5. Himerni polipeptid u skladu s patentnim zahtjevom 1 do 4, naznačen time, da se vezna domena veže na intracelularni epitop GPCR-a, poželjno na unutarstanični epitop koji se nalazi na mjestu vezanja proteina za silaznu signalizaciju, poželjnije unutarstanični epitop koji se nalazi u mjestu vezanju G proteina.
6. Himerni polipeptid u skladu s patentnim zahtjevima 1 do 5, naznačen time, da je varijabilna domena imunoglobulina izvedena iz antitijela teškog lanca, pri čemu je poželjno da imunoglobulinska pojedinačna varijabilna domena sadržava VHH sekvencu ili sekvencu nanotijela.
7. Himerni polipeptid u skladu s patentnim zahtjevom 1 do 6, naznačen time, da je GPCR prirodno prisutan GPCR ili varijanta prirodnog GPCR-a ili skraćenog oblika prirodnog GPCR-a, pri čemu je GPCR poželjno odabran iz skupine koja sadržava GPCR iz obitelji glutamata GPCR, GPCR iz obitelji Rodopsin GPCR-a, GPCR iz adhezijske obitelji GPCR-a, GPCR „frizzled" / ,,Taste2" obitelji GPCR-a i GPCR Secretinove obitelji GPCR-a.
8. Kompleks, naznačen time, da sadržava himerni polipeptid u skladu s patentnim zahtjevima 1 do 7 i ligand za receptor.
9. Himerni polipeptid u skladu s patentnim zahtjevima 1-7, ili kompleks u skladu s patentnim zahtjevom 8, naznačen time, da je kristalne strukture.
10. Molekula nukleinske kiseline, naznačen time, da sadržava sekvencu nukleinske kiseline koja kodira himerni polipeptid u skladu s patentnim zahtjevima 1 do 7.
11. Stanica domaćina, poželjno bakterijska stanica, stanica kvasca, stanica sisavaca ili stanica insekata, naznačena time, da sadržava sekvencu nukleinske kiseline iz patentnog zahtjeva 10.
12. Membranska kompozicija, naznačena time, da sadržava himerni polipeptid u skladu s patentnim zahtjevima 1 do 7 ili kompleks u skladu s patentnim zahtjevom 8.
13. Postupak za prikazivanje GPCR-a u aktivnoj ili neaktivnoj konformaciji na staničnoj površini ili u određenoj staničnoj membranskoj frakciji stanice domaćina, naznačen time, da postupak obuhvaća korake pripreme stanice domaćina u skladu s patentnim zahtjevom 11 i uzgajanja stanice pod pogodnim uvjetima za ekspresiju himerni polipeptid.
14. Postupak identificiranja spojeva selektivnih na konformaciju, poželjno agonističkih spojeva kao što je agonistička mala molekula ili protutijelo, naznačen time, da postupak obuhvaća korake i. pripremu himernog polipeptida u skladu s patetnim zahtjevima 1 do 7, i ii. pripremu ispitivanog spoja i iii. procjena selektivnog vezanja ispitivanog spoja na GPCR koji je sadržan u himernom polipeptidu.
15. Uporaba himernog polipeptida u skladu s patetnim zahtjevima 1 do 7 ili kompleksa u skladu s patentnim zahtjevom 8, naznačena time, da kristalizira i/ili otapa strukturu GPCR-a ili hvata GPCR u funkcionalnom konformacijskom stanju.
HRP20171777TT 2013-01-30 2014-01-30 Novi himerni polipeptidi za potrebe probira i otkrivanja lijekova HRP20171777T1 (hr)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US201361758518P 2013-01-30 2013-01-30
EP13154552 2013-02-08
EP13187265 2013-10-03
EP14702547.2A EP2951201B1 (en) 2013-01-30 2014-01-30 Novel chimeric polypeptides for screening and drug discovery purposes
PCT/EP2014/051845 WO2014118297A1 (en) 2013-01-30 2014-01-30 Novel chimeric polypeptides for screening and drug discovery purposes

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US (1) US9593157B2 (hr)
EP (1) EP2951201B1 (hr)
JP (1) JP6443344B2 (hr)
CN (2) CN112858672A (hr)
AU (1) AU2014211355B2 (hr)
CA (1) CA2899693C (hr)
DK (1) DK2951201T3 (hr)
ES (1) ES2654675T3 (hr)
HR (1) HRP20171777T1 (hr)
HU (1) HUE035315T2 (hr)
NO (1) NO2951201T3 (hr)
PL (1) PL2951201T3 (hr)
PT (1) PT2951201T (hr)
WO (1) WO2014118297A1 (hr)

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PT2951201T (pt) 2018-01-09
EP2951201B1 (en) 2017-10-18
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AU2014211355B2 (en) 2017-07-13
CA2899693C (en) 2023-03-14
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NO2951201T3 (hr) 2018-03-17
HUE035315T2 (en) 2018-05-02
EP2951201A1 (en) 2015-12-09
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