Folate derivatives, useful in particular in the context of the folate assay

• US 9,926,323 B2
• Filed: 12/27/2013
• Issued: 03/27/2018
• Est. Priority Date: 12/27/2012
• Status: Active Grant

• Current Assignee: Biomerieux B.V. (Compagnie Merieux Alliance SAS)
• Original Assignee: Biomerieux B.V. (Compagnie Merieux Alliance SAS)
• Inventors: Guo, Yuping, Cheucle, Sylvie
• Primary Examiner(s): Haq, Shafiqul
• Application Number:
  US14/655,258
• Publication Number:
  US 20150355201A1
• Time in Patent Office: 1,551 Days
• CPC Class Codes: C07D 475/04 with a nitrogen atom directly attached in position 2
• CPC Class Codes: G01N 33/82 involving vitamins or their receptors

Abstract

Use of a folate derivative to assay in vitro the folate in a sample such as a biological sample.

Claims

1. A method of in vitro assaying of folate(s) in a sample, the method comprising:

(a) Contacting the sample with a folate binding partner in the presence of a folate conjugate decarboxylated in the α

position and suitable for emitting a signal in a non-radioisotopic competition immunoassay, the folate conjugate corresponding to general formula (III);

2. The method according to claim 1, wherein X is a linear aliphatic saturated hydrocarbon chain containing from 2 to 7 carbon atoms.

6. The method according to claim 2, wherein the hydrocarbon chain contains from 3 to 5 carbon atoms.

7. The method of claim 1 wherein prior to step (a), the sample is treated to dissociate the folate(s) from other molecules in the sample.

8. The method of claim 1 wherein Y′

is of general formula (IV).

9. The method of claim 1 wherein R1 is NH or O.

10. The method of claim 1 wherein R1 is NH.

11. The method of claim 1 wherein the compound of formula III is selected from the group consisting of

12. The method of claim 1 wherein the binding partner is selected from the group consisting of antibodies, antibody fragments, nanofitins, folates receptor or protein;

wherein said binding partner binds folates(s).

13. The method of claim 1 wherein M is selected from an enzyme, a chromophore, a fluorescent molecule or an electrochemiluminescent salt.

14. The method of claim 13 wherein M is selected from the group consisting of horseradish peroxidase, alkaline phopatase, β

-galactosidase, glucose-6-phosphate dehydrogenase, Alexa, phycocyanins, and organometallic derivatives based on acridinium and ruthenium.

3. A process of in vitro non-radioisotopic competition immunoassay of folate(s) in a biological sample, the process comprising:

a) contacting the biological sample with at least one folate binding partner suitable to in the presence of a with at least one conjugate corresponding to the general formula (III);

5. The process according to claim 3, wherein referencing a calibration curve comprises comparing the measured concentration of the folate(s) with a threshold value corresponding to a predetermined concentration of the folate(s).

4. A method for in vitro determination of folate deficiency in a human or animal patient, the method comprising:

a) assaying folate(s) in a biological sample by

i. contacting the biological sample with at least one folate binding partner suitable to in the presence of a with at least one conjugate corresponding to the general formula (III);