Splice variants for human 5-HT4 serotonin receptor and their applications, in particular for screening

• US 6,506,580 B1
• Filed: 09/19/2000
• Issued: 01/14/2003
• Est. Priority Date: 11/28/1997
• Status: Expired due to Fees

• Current Assignee: Institut National De La Sante Et De La Recherche Medicale (Inserm) (Allianz Global Investors GmbH)
• Original Assignee: Institut National De La Sante Et De La Recherche Medicale (Inserm) (Allianz Global Investors GmbH)
• Inventors: Hoebeke, Johan, Blondel, Olivier, Dahmoune, Yamina, Gastineau, Monique, Langlois, Michel, Fischmeister, Rodolphe
• Primary Examiner(s): ULM, JOHN D
• Application Number:
  US09/555,313
• Time in Patent Office: 847 Days
• Field of Search: 435/69.1, 435/252.3, 435/320.1, 435/7.1, 435/7.2, 435/7.21, 536/23.5
• US Class Current: 435/69.1
• CPC Class Codes: A61K 31/445 Non condensed piperidines, e.g. piperocaine
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: C07K 14/70571 for neuromediators, e.g. serotonin receptor, dopamine receptor

Abstract

The invention concerns an isolated polypeptide constitutive of splice variants for human serotonin receptor whereof the amino acid sequence is selected among the sequence SEQ ID No. 2 of the 5-HT_4(c) polypeptide variant or the sequence SEQ ID No. 4 of the 5-HT_4(d) polypeptide variant, or any polypeptide fragment or biologically active derivative thereof. The invention also concerns the inverse agonist effect of ML 10375 on 5-HT4(c) and 5-HT4(d) receptors.

Claims

1. Isolated nucleotide sequence chosen from the sequence SEQ ID No. 1, the sequence SEQ ID No. 3, the nucleotide sequences derived from the sequence SEQ ID No. 1 or from the sequence SEQ ID No. 3 due to the degeneracy of the genetic code, to mutation, to deletion or to insertion, and the nucleotide sequences which are capable of specifically hybridizing with the sequence SEQ ID No. 1 or the sequence SEQ ID No. 3.

2. Cloning and/or expression vector containing a nucleotide sequence according to claim 1.

3. Host cell transformed with a vector according to claim 2.

4. Method for producing a biologically active recombinant polypeptide 5-HT_4(c) or 5-HT_4(d), fragment or derivative of this polypeptide, in which cells transformed according to claim 3 are cultured under conditions which allow the expression of a polypeptide of sequence SEQ ID No. 2 or SEQ ID No. 4, or any biologically active fragment or derivative of this polypeptide, and said biologically active polypeptide, fragment or derivative of this polypeptide is recovered.

5. A method for evaluating the pharmacological properties of the compounds which are capable of binding to the polypeptide 5-HT_4(d), termed receptor 5-HT_4(d) ligands, comprising the steps of:

a) culturing host cells according to claim 3 which express the polypeptide 5-HT_4(d) ligand; and

b) evaluating the capacity of the ligand for modifying signal transduction.

6. The method according to claim 5 in which the capacity of the ligand for modulating signal transduction is evaluated by determining the intracellular cAMP concentration or adenylate cyclase activity.