DK2327985T3 - Heterooligomere T1R-smagsreceptorer og cellelinjer, der udtrykker receptorerne, samt anvendelse deraf til identificering af smagsforbindelser - Google Patents

Heterooligomere T1R-smagsreceptorer og cellelinjer, der udtrykker receptorerne, samt anvendelse deraf til identificering af smagsforbindelser Download PDF

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DK2327985T3
DK2327985T3 DK10180331.0T DK10180331T DK2327985T3 DK 2327985 T3 DK2327985 T3 DK 2327985T3 DK 10180331 T DK10180331 T DK 10180331T DK 2327985 T3 DK2327985 T3 DK 2327985T3
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DK10180331.0T
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Mark T Zoller
Xiaodong Li
Lena Staszewski
Shawn O'connell
Sergey Zozulya
Jon Elliot Adler
Hong Xu
Fernando Echeverri
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Senomyx Inc
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Priority claimed from US09/897,427 external-priority patent/US6955887B2/en
Priority claimed from US10/035,045 external-priority patent/US7241880B2/en
Application filed by Senomyx Inc filed Critical Senomyx Inc
Priority claimed from EP02761016A external-priority patent/EP1412750B1/en
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Publication of DK2327985T3 publication Critical patent/DK2327985T3/da

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    • 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
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
    • G01N33/5041—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects involving analysis of members of signalling pathways
    • 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
    • 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
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • 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
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
    • G01N33/5023—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects on expression patterns
    • 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
    • G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
    • 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
    • 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

Claims (15)

1. In vitro-fremgangsmåde til identificering af en celle, der er potentielt følsom for søde smagsstimuli, hvilken fremgangsmåde omfatter: (a) påvisning af ekspression af et TlR2-polypeptid og/eller en nukleinsyre, der koder for T1R2-polypeptidet af cellen, hvor TlR2-polypeptidet (i) kodes af en nukleinsyresekvens, der omfatter SEQ ID NO: 10, eller en sekvens, der har mindst 90 % sekvensidentitet med SEQ ID NO: 10; eller (ii) har mindst 90 % sekvensidentitet med TlR2-polypeptidet ifølge SEQ ID NO: 6; og (b) påvisning af ekspression af et TlR3-polypeptid og/eller en nukleinsyre, der koder for TlR3-polypeptidet af cellen, hvilket TlR3-polypeptid (i) kodes af en nukleinsyresekvens, der omfatter SEQ ID NO: 9 eller en sekvens, der har mindst 90 % sekvensidentitet med SEQ ID NO: 9; eller (ii) har mindst 90 % sekvensidentitet med TlR3-polypeptidet ifølge SEQ ID NO: 7.
2. Fremgangsmåde ifølge krav 1, hvor trin (a) omfatter etablering af kontakt mellem cellen og mindst én probe, der påviser ekspression af T1R2-polypeptidet og/eller nukleinsyren, der koder for T1R2-polypeptidet af cellen, og hvor trin (b) omfatter etablering af kontakt mellem cellen og mindst én probe, der påviser ekspression af TlR3-polypeptidet og/eller nukleinsyren, der koder for TlR3-polypeptidet af cellen.
3. Fremgangsmåde ifølge krav 1 eller krav 2, der yderligere omfatter et trin (c) til bekræftelse af, at cellen, der i trin (a) og trin (b) er identificeret at udtrykke T1R2 og T1R3, er følsom for søde smagsstimuli.
4. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 3, der yderligere omfatter anvendelse af den identificerede celle i en screeningsanalyse med højt gennemløb, der påviser forbindelser, der specifikt binder, aktiverer eller modulerer aktiveringen af en receptor, der omfatter T1R2- og TlR3-polypeptiderne, eller der modulerer bindingen eller aktiveringen af en receptor, der omfatter T1R2- og TlR3-polypeptiderne, ved hjælp af en anden forbindelse.
5. Fremgangsmåde ifølge et hvilket som helst af ovennævnte krav, hvor cellen er en mammaliacelle.
6. Fremgangsmåde ifølge krav 5, hvor cellen er en abecelle eller en human celle.
7. In vitro-fremgangsmåde til screening for en forbindelse, der formodes at blokere eller aktivere sød smagssignalering, hvilken fremgangsmåde omfatter trinene: (a) etablering af kontakt mellem celler og en eller flere forbindelser, hvor cellerne udtrykker en heterooligomer TlR2/TlR3-smagsreceptor; og (b) påvisning af, om den ene eller de flere forbindelser specifikt binder til den heterooligomere T1R2/T1R3-smagsreceptor og/eller specifikt aktiverer den heterooligomere TlR2/TlR3-smagsreceptor og på baggrund deraf identificering af den ene eller de flere forbindelser som forbindelser, der formodes at blokere eller aktivere sød smagssignalering, hvor den heterooligomere TlR2/TlR3-smagsreceptor, der udtrykkes af cellerne, omfatter mindst ét TlR2-polypeptid, der (i) kodes af en nukleinsyresekvens, der omfatter SEQ ID NO: 10 eller en sekvens, der har mindst 90 % sekvensidentitet med SEQ ID NO: 10; eller (ii) har mindst 90 % sekvensidentitet med TlR2-polypeptidet ifølge SEQ ID NO: 6, og hvor den heterooligomere TlR2/TlR3-smagsreceptor, der udtrykkes af cellerne, omfatter mindst ét TlR3-polypeptid, der (i) kodes af en nukleinsyresekvens, der omfatter SEQ ID NO: 9 eller en sekvens, der har mindst 90 % sekvensidentitet med SEQ ID NO: 9; eller (ii) har mindst 90 % sekvensidentitet med TlR3-polypeptidet ifølge SEQ ID NO: 7.
8. In vitro-fremgangsmåde til screening for en forbindelse, der formodes at modulere sød smagssignalering, hvilken fremgangsmåde omfatter trinene: (a) etablering af kontakt mellem celler og en eller flere forbindelser, hvor cellerne udtrykker en heterooligomer TlR2/TlR3-smagsreceptor; og (b) påvisning af, om den ene eller de flere forbindelser påvirker bindingen af en anden forbindelse til den heterooligomere TlR2/TlR3-smagsreceptor og/eller modulerer aktiveringen af den heterooligomere TlR2/TlR3-smagsreceptor ved hjælp af en anden forbindelse og på baggrund deraf identificering af den ene eller de flere forbindelser som forbindelser, der formodes at modulere sød smagssignalering, hvor den heterooligomere TlR2/TlR3-smagsreceptor, der udtrykkes af cellerne, omfatter mindst ét TlR2-polypeptid, der (i) kodes af en nukleinsyresekvens, der omfatter SEQ ID NO: 10 eller en sekvens, der har mindst 90 % sekvensidentitet med SEQ ID NO: 10; eller (ii) har mindst 90 % sekvensidentitet med TlR2-polypeptidet ifølge SEQ ID NO: 6, og hvor den heterooligomere TlR2/TlR3-smagsreceptor, der udtrykkes af cellerne, omfatter mindst ét TlR3-polypeptid, der (i) kodes af en nukleinsyresekvens, der omfatter SEQ ID NO: 9 eller en sekvens, der har mindst 90 % sekvensidentitet med SEQ ID NO: 9; eller (ii) har mindst 90 % sekvensidentitet med TlR3-polypeptidet ifølge SEQ ID NO: 7.
9. Fremgangsmåde ifølge krav 7 eller krav 8, hvor cellerne omfatter isolerede eller dyrkede mammaliaceller.
10. Fremgangsmåde ifølge et hvilket som helst af kravene 7 til 9, hvor den heterooligomere TlR2/TlR3-smagsreceptor, der udtrykkes af cellerne, binder specifikt til en ligand, der binder specifikt til en endogen (vildtype) human heterooligomer T1R2/TlR3-receptor, der omfatter mindst ét endogent TlR2-polypeptid og mindst ét endogent T1R3- polypeptid, og hvor mindst det ene af det udtrykte T1R2-polypeptid eller det udtrykte TlR3-polypeptid udtrykkes fra et gen, der er endogent for cellerne.
11. Fremgangsmåde ifølge et hvilket som helst af kravene 7 til 10, der yderligere indbefatter en smagstest til bestemmelse af virkningen af den identificerede forbindelse på sød smag og på baggrund af resultatet af smagstesten identificering af forbindelsen som en forbindelse, der blokerer, aktiverer eller modulerer sød smagssignalering.
12. Fremgangsmåde ifølge krav 7 eller krav 8, hvor det udtrykte TlR2-polypeptid udtrykkes fra et gen, der er endogent for cellerne.
13. Fremgangsmåde ifølge krav 7 eller krav 8, hvor det udtrykte TlR3-polypeptid udtrykkes fra et gen, der er endogent for cellerne.
14. Fremgangsmåde ifølge et hvilket som helst af kravene 7 til 13, hvor cellerne er humane celler eller abeceller.
15. Fremgangsmåde ifølge et hvilket som helst af kravene 7 til 14, der omfatter en scenningsmetode med højt gennemløb.
DK10180331.0T 2001-06-26 2002-06-26 Heterooligomere T1R-smagsreceptorer og cellelinjer, der udtrykker receptorerne, samt anvendelse deraf til identificering af smagsforbindelser DK2327985T3 (da)

Applications Claiming Priority (10)

Application Number Priority Date Filing Date Title
US30043401P 2001-06-26 2001-06-26
US09/897,427 US6955887B2 (en) 2001-03-30 2001-07-03 Use of T1R hetero-oligomeric taste receptor to screen for compounds that modulate taste signaling
US30474901P 2001-07-13 2001-07-13
US31049301P 2001-08-08 2001-08-08
US33177101P 2001-11-21 2001-11-21
US33947201P 2001-12-14 2001-12-14
US10/035,045 US7241880B2 (en) 2001-01-03 2002-01-03 T1R taste receptors and genes encoding same
US37209002P 2002-04-15 2002-04-15
US37414302P 2002-04-22 2002-04-22
EP02761016A EP1412750B1 (en) 2001-06-26 2002-06-26 T1r2-t1r3 hetero-oligomeric sweet taste receptors and cell lines that express said receptors and use thereof for identification of sweet taste compounds

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US20050287517A1 (en) 2005-12-29
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US20130244254A1 (en) 2013-09-19
US8119359B2 (en) 2012-02-21
US8809000B2 (en) 2014-08-19
US10591464B2 (en) 2020-03-17
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US20150024407A1 (en) 2015-01-22
US10088472B2 (en) 2018-10-02

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