CA2553769A1 - Npc1l1 (npc3) and methods of identifying ligands thereof - Google Patents

Npc1l1 (npc3) and methods of identifying ligands thereof Download PDF

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Publication number
CA2553769A1
CA2553769A1 CA002553769A CA2553769A CA2553769A1 CA 2553769 A1 CA2553769 A1 CA 2553769A1 CA 002553769 A CA002553769 A CA 002553769A CA 2553769 A CA2553769 A CA 2553769A CA 2553769 A1 CA2553769 A1 CA 2553769A1
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CA
Canada
Prior art keywords
azetidinone
glucuronide
substituted
value
npc1l1
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Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
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CA002553769A
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French (fr)
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CA2553769C (en
Inventor
Margarita Garcia-Calvo
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Merck Sharp and Dohme LLC
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Individual
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Publication date
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Publication of CA2553769A1 publication Critical patent/CA2553769A1/en
Application granted granted Critical
Publication of CA2553769C publication Critical patent/CA2553769C/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING 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
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/60—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving cholesterol
    • 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
    • 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
    • G01N33/5076—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 involving cell organelles, e.g. Golgi complex, endoplasmic reticulum
    • 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

Abstract

The present invention provides human, rat and mouse NPCIL1 polypeptides and polynucleotides encoding the polypeptides. Methods for detecting ligands which bind to NPClL1 and block intestinal cholesterol absorption are provided. Also included is a method of identifying ligands which bind to NPCILI using membranes derived from brush border membrane preparations. Compounds that bind to NPCILI can be used for inhibiting intestinal cholesterol absorption in a subject.

Claims (20)

1. A method for identifying a ligand of NPC 1 L1 comprising:
contacting human NPC1L1 with a detectably labeled substituted 2-azetidinone glucuronide and a candidate compound; and determining whether said candidate compound binds to human NPC1L1;
wherein binding of said candidate compound to human NPC1L1 modulates binding of said detectably labeled substituted 2-azetidinone glucuronide to human NPC1L1, wherein the detectably labeled substituted 2-azetidinone glucuronide has a binding affinity KD value for human NPC1L1 that is 200nM or lower, and wherein said modulation indicates that the candidate compound is a ligand that binds to human NPC1L1.
2. The method of claim 1, wherein the KD value is 100nM or lower.
3. The method of claim 1, wherein the KD value is 50nM or lower.
4. The method of claim 1, wherein the KD value is 20nM or lower.
5. The method of claim 1, wherein the KD value is 10nM or lower.
6. The method of claim 1, wherein the substituted 2-azetidinone-glucuronide is selected from the group consisting of a compound of Formula I and a compound of Formula II.
7. The method of claim 6, wherein the substituted 2-azetidinone-glucuronide comprises a detectable label from the group consisting of 35S and 125I.
S. The method of claim 7, wherein the detectable label is 35S.
9. The method of claim 6, wherein the substituted 2-azetidinone-glucuronide is a compound of Formula II, wherein R9 comprises an -SO2- group.
10. The method of claim 9, wherein the substituted 2-azetidinone-glucuronide of Formula II is labeled with 35S.
11. A method for identifying a ligand of NPC1L1 comprising:
contacting human NPC1L1 with a detestably labeled substituted 2-azetidinone glucuronide of Formula II and a candidate compound; and determining whether said candidate compound binds to human NPC1L1;
wherein binding of said candidate compound to human NPC1L1 modulates binding of said detestably labeled substituted 2-azetidinone glucuronide of Formula II
to human NPC1L1, and wherein said modulation indicates that the candidate compound is a ligand that binds to human NPC1L1.
12. The method of claim 11, wherein R9 of the detestably labeled substituted 2-azetidinone glucuronide of Formula II comprises an -SO2- group.
13. The method of claim 11, wherein the detestably labeled substituted 2-azetidinone glucuronide of Formula II is labeled with 35S.
14. The method of claim 11, wherein the detestably labeled substituted 2-azetidinone glucuronide of Formula II has a binding affinity K D value for human NPC1L1 that is 200nM or lower.
15. The method of claim 14, wherein the K D value is 100nM or lower.
16. The method of claim 14, wherein the K D value is 50nM or lower.
17. The method of claim 14, wherein the K D value is 20nM or lower.
18. The method of claim 14, wherein the K D value is 10nM or lower.
19. The method of claim 1 wherein the detestably labeled substituted 2-azetidinone glucuronide is labeled with 35S.
20. The method of claim 1 wherein the detestably labeled substituted 2-azetidinone glucuronide is 35S-labeled compound 2.
CA2553769A 2004-01-16 2005-01-14 Npc1l1 (npc3) and methods of identifying ligands thereof Expired - Fee Related CA2553769C (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US53734104P 2004-01-16 2004-01-16
US60/537,341 2004-01-16
PCT/US2005/001469 WO2005069900A2 (en) 2004-01-16 2005-01-14 Npc1l1 (npc3) and methods of identifying ligands thereof

Publications (2)

Publication Number Publication Date
CA2553769A1 true CA2553769A1 (en) 2005-08-04
CA2553769C CA2553769C (en) 2011-01-04

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Family Applications (1)

Application Number Title Priority Date Filing Date
CA2553769A Expired - Fee Related CA2553769C (en) 2004-01-16 2005-01-14 Npc1l1 (npc3) and methods of identifying ligands thereof

Country Status (6)

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US (1) US7901893B2 (en)
EP (1) EP1723414A4 (en)
JP (1) JP4590417B2 (en)
CA (1) CA2553769C (en)
MX (1) MXPA06008124A (en)
WO (1) WO2005069900A2 (en)

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JP2008543837A (en) * 2005-06-15 2008-12-04 メルク エンド カムパニー インコーポレーテッド Anti-hypercholesterolemic compound
UY29607A1 (en) 2005-06-20 2007-01-31 Astrazeneca Ab CHEMICAL COMPOUNDS
AR057072A1 (en) 2005-06-22 2007-11-14 Astrazeneca Ab CHEMICAL COMPOUNDS DERIVED FROM 2-AZETIDINONE, PHARMACEUTICAL FORMULATION AND A COMPOUND PREPARATION PROCESS
SA06270191B1 (en) 2005-06-22 2010-03-29 استرازينيكا ايه بي Novel 2-Azetidinone Derivatives as Cholesterol Absorption Inhibitors for the Treatment of Hyperlipidaemic Conditions
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EP2581094A4 (en) * 2010-06-10 2015-04-08 Adbiotech Co Ltd Composition for inhibiting hyperlipidemia and obesity by inhibiting intestinal absorption of cholesterol
US10377815B2 (en) 2015-05-05 2019-08-13 Albert Einstein College Of Medicine Bispecific antibodies and fusion proteins that bind to EBOV and NPC1

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Also Published As

Publication number Publication date
JP2007526998A (en) 2007-09-20
WO2005069900B1 (en) 2006-08-03
CA2553769C (en) 2011-01-04
MXPA06008124A (en) 2007-01-26
WO2005069900A3 (en) 2006-06-08
JP4590417B2 (en) 2010-12-01
US7901893B2 (en) 2011-03-08
US20100009461A1 (en) 2010-01-14
EP1723414A2 (en) 2006-11-22
EP1723414A4 (en) 2008-03-26
WO2005069900A2 (en) 2005-08-04

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