Use of quinolinone derivatives as a research tool

• US 8,957,091 B2
• Filed: 11/15/2011
• Issued: 02/17/2015
• Est. Priority Date: 11/16/2010
• Status: Expired due to Fees

• Current Assignee: Centre National De La Recherche Scientifique
• Original Assignee: Centre National De La Recherche Scientifique
• Inventors: Ruat, Martial, Faure, Helene, Gorojankina, Tatiana
• Primary Examiner(s): Seaman, D M
• Application Number:
  US13/885,098
• Publication Number:
  US 20130236912A1
• Time in Patent Office: 1,190 Days
• Field of Search: 514/312, 546/159
• US Class Current: 514/312
• CPC Class Codes: G01N 2333/726 G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH
• CPC Class Codes: G01N 2500/04 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)
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds
• CPC Class Codes: G01N 33/574 for cancer
• CPC Class Codes: G01N 33/68 involving proteins, peptides or amino acids involving lipoproteins G01N33/92

Abstract

The present invention relates to the use of quinolinone derivatives of general formula (I), ligands of the Smo receptor or of related receptors which target a binding site of the receptor which is different from the known ligand binding sites, as research tools for identifying modulators of the Smo receptor or of related receptors, characterizing the Hedgehog signalling pathway and diagnosis; the invention also relates to kits containing said derivatives of general formula (I).

Claims

1. A method of activating the Hedgehog signaling pathway, comprising bringing a Smoothened receptor and at least one compound of general formula (I) below into contact:

2. The method as claimed in claim 1, wherein the compound of general formula (I) is selected from the group consisting of

propyl 4-(1-hexyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxamido)benzoate (compound 1);

ethyl 4-(1-benzyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxamido)benzoate (compound

     2);

ethyl 4-(1-hexyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxamido)benzoate (compound

     3);

4-(1-hexyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxamido)benzoic acid (compound

     4);

butyl 4-(1-hexyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxamido)benzoate (compound

     5);

tert-butyl 4-(1-hexyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxamido)benzoate (compound

     6);

benzyl 4-(1-hexyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxamido)benzoate (compound

     7);

N-(4-(butylcarbamoyl)phenyl-1-hexyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxamide (compound

     8); and

isopropyl 4-(1-hexyl-4-hydroxy-2-oxo-1,2-dihydroquinoline-3-carboxamido)benzoate (compound

     9).

3. A method for screening for and/or identifying ligands of the Smo2 binding site of the Smoothened receptor, comprising the steps of:

a) bringing the Smoothened receptor and at least one compound of general formula (I) as defined in claim 1 into contact so as to obtain an [Smo-compound of general formula (I)] complex;

b) bringing the Smoothened receptor, said compound of general formula (I) and a test molecule into contact;

c) detecting an interaction between said Smoothened receptor and said test molecules by comparing the Smoothened receptor recovered in step b) with the [Smo-compound of general formula (I)] complex; and

d) selecting said test molecules for which an interaction with the Smoothened receptor is measured.

4. The method as claimed in claim 3, wherein step a) is carried out in the liquid phase and in that said step comprises the following steps:

a) bringing the Smoothened receptor and at least one compound of general formula (I) into contact so as to obtain an [Smo-compound of general formula (I)] complex;

b) bringing the Smoothened receptor, said compound of general formula (I) and a test molecule into contact;

c) recovering said Smoothened receptor possibly bound to one or more test molecules and/or to said compound of general formula (I);

d) detecting an interaction between said Smoothened receptor and said test molecules by comparing the Smoothened receptor recovered in step c) with the [Smo-compound of general formula (I)] complex; and

e) selecting said test molecules for which an interaction is measured.

5. The method as claimed in claim 3, wherein step a) is carried out on the solid support.

6. The method as claimed in claim 5, wherein the method makes it possible to identify Smoothened receptor ligands which do not bind to the Smo2 binding site.

7. A method for identifying Smoothened receptor agonists, wherein the method comprises the steps of the methods as claimed in claim 3 and the additional steps:

of bringing said selected molecule into contact with a cell which exhibits a cellular response following activation of the Smoothened receptor and

of selecting the molecules capable of inducing said cellular response of said cell.

8. A method for identifying a Smoothened receptor antagonist molecule, comprising the following steps:

a) culturing cells which exhibit a cellular response following the activation of the Smoothened receptor with at least one compound of general formula (I) as defined in claim 1 so as to induce said cellular response;

b) bringing the cells obtained at the end of step a) into contact with a test molecule;

c) selecting the molecules which induce an inhibition of said cellular response of said cells.

9. A method for identifying an antagonist molecule molecules capable of inducing a potentiation of Smoothened receptor activation, comprising the following steps:

a) culturing cells which exhibit a cellular response following the activation of the Smoothened receptor with at least one compound of general formula (I) as defined in claim 1 so as to induce said cellular response;

b) bringing the cells obtained at the end of step a) into contact with a test molecule;

c) selecting the molecules which induce an increase in the cellular response of the cells which exhibit a cellular response following the activation of the Smoothened receptor.

10. A kit comprising at least the functional Smoothened receptor and at least one compound of general formula (I) as defined in claim 1.

11. A method for identifying cells, such as tumor cells, expressing the Smoothened receptor, comprising the steps of:

a) bringing a test cell into contact with a labeled compound of general formula (I) as defined in claim 1;

b) cleaning the cells in order to remove said labeled compound of general formula (I) which has not bound any receptor of said test cell;

c) detecting the labeled cells.