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1. WO2020065338 - CELL-BASED CLOSTRIDAL NEUROTOXIN ASSAYS

Publication Number WO/2020/065338
Publication Date 02.04.2020
International Application No. PCT/GB2019/052734
International Filing Date 27.09.2019
IPC
C12Q 1/37 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
34involving hydrolase
37involving peptidase or proteinase
G01N 33/50 2006.01
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48Biological material, e.g. blood, urine; Haemocytometers
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
G01N 33/542 2006.01
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48Biological material, e.g. blood, urine; Haemocytometers
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
536with immune complex formed in liquid phase
542with steric inhibition or signal modification, e.g. fluorescent quenching
G01N 33/569 2006.01
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48Biological material, e.g. blood, urine; Haemocytometers
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
569for microorganisms, e.g. protozoa, bacteria, viruses
CPC
A61K 47/02
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
47Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
02Inorganic compounds
A61K 47/183
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
47Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
06Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
16containing nitrogen, ; e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
18Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
183Amino acids, e.g. glycine, EDTA or aspartame
A61K 9/0019
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
9Medicinal preparations characterised by special physical form
0012Galenical forms characterised by the site of application
0019Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
C12N 15/10
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12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
10Processes for the isolation, preparation or purification of DNA or RNA
C12N 15/1086
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
10Processes for the isolation, preparation or purification of DNA or RNA
1034Isolating an individual clone by screening libraries
1086Preparation or screening of expression libraries, e.g. reporter assays
C12N 2510/00
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
2510Genetically modified cells
Applicants
  • IPSEN BIOPHARM LIMITED [GB]/[GB]
  • INSTITUTE OF MOLECULAR AND CELL BIOLOGY, BIOMEDICAL SCIENCES INSTITUTES [SG]/[SG]
Inventors
  • FOSTER, Keith
  • BEARD, Matthew
  • YEO, Jeremy Changyu
  • BARD, Frederic Andre Jean
  • TAY, Pei Ling Felicia
Agents
  • HOBSON, David James
Priority Data
1815870.928.09.2018GB
Publication Language English (EN)
Filing Language English (EN)
Designated States
Title
(EN) CELL-BASED CLOSTRIDAL NEUROTOXIN ASSAYS
(FR) ESSAIS DE NEUROTOXINE CLOSTRIDIALE À BASE DE CELLULES
Abstract
(EN)
The present invention is directed to a method for identifying a gene that regulates clostridial neurotoxin activity, the method comprising: a. providing a sample of human neuronal cells expressing a polypeptide that comprises a C-terminal detectable label, wherein the polypeptide is cleavable by a clostridial neurotoxin; b. altering expression of a target gene of the cells; c. contacting the cells with the clostridial neurotoxin; d. measuring an amount of C-terminal detectable label, thereby quantifying clostridial neurotoxin activity; and e. identifying the target gene as a regulator of clostridial neurotoxin activity when the quantified clostridial neurotoxin activity is different to the quantified clostridial neurotoxin activity when expression of the target gene is unaltered; or f. identifying that the target gene is not a regulator of clostridial neurotoxin activity when the quantified clostridial neurotoxin activity is equivalent to the quantified clostridial neurotoxin activity when expression of the target gene is unaltered. Also provided are related methods for identifying an agent that regulates clostridial neurotoxin activity, as well as human neuronal cells, nucleotides, vectors, polypeptides, kits, and compositions suitable for use in the methods of the invention.
(FR)
La présente invention concerne un procédé d'identification d'un gène qui régule l'activité d'une neurotoxine clostridiale, le procédé consistant à a. : utiliser un échantillon de cellules neuronales humaines exprimant un polypeptide qui comprend une étiquette détectable au niveau de l'extrémité C, le polypeptide étant clivable par une neurotoxine clostridiale ; b. modifier l'expression d'un gène cible des cellules ; c. mettre en contact les cellules avec la neurotoxine clostridiale ; d. mesurer une quantité d'étiquette détectable au niveau de l'extrémité C, ce qui permet de quantifier l'activité de la neurotoxine clostridiale ; et e. identifier le gène cible en tant que régulateur de l'activité de la neurotoxine clostridiale lorsque l'activité quantifiée de la neurotoxine clostridiale est différente de l'activité quantifiée de la neurotoxine clostridiale lorsque l'expression du gène cible est inchangée ; ou f. identifier le fait que le gène cible n'est pas un régulateur de l'activité de neurotoxine clostridiale lorsque l'activité quantifiée de la neurotoxine clostridiale est équivalente à l'activité quantifiée de la neurotoxine clostridiale lorsque l'expression du gène cible est inchangée. L'invention concerne également des procédés associés pour identifier un agent qui régule l'activité de la neurotoxine clostridiale, ainsi que des cellules neuronales humaines, des nucléotides, des vecteurs, des polypeptides, des kits et des compositions appropriés pour une utilisation dans les procédés de l'invention.
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