Processes using a human serotonin receptor (5-HT.sub.4B)

• US 5,985,585 A
• Filed: 06/15/1995
• Issued: 11/16/1999
• Est. Priority Date: 11/03/1992
• Status: Expired due to Term

• Current Assignee: Lundbeck Research USA, Inc. (Lundbeck Foundation)
• Original Assignee: Synaptic Pharmaceutical Corporation (Lundbeck Foundation)
• Inventors: Bard, Jonathan A., Branchek, Theresa, Weinshank, Richard L.
• Primary Examiner(s): Allen, Marianne P.
• Application Number:
  US08/157,185
• Time in Patent Office: 1,615 Days
• Field of Search: 435/7.1, 435/7.2, 435/7.21, 435/325, 435/69.1, 435/356, 435/357, 435/358, 435/365
• US Class Current: 435/7.21
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/00 Genetically modified animals
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2217/072 maintaining or altering function, i.e. knock in
• CPC Class Codes: A01K 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: A01K 2227/10 Mammal
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/03 Animal model, e.g. for test or diseases
• CPC Class Codes: A01K 2267/0356 Animal model for processes and diseases of the central nervous system, e.g. stress, learning, schizophrenia, pain, epilepsy
• CPC Class Codes: A01K 2267/0375 Animal model for cardiovascular diseases
• 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: C07K 14/70571 for neuromediators, e.g. serotonin receptor, dopamine receptor
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic

Abstract

The invention provides for processes for identifying chemical compounds which specifically bind to a human 5-HT_4B having the amino acid sequence of SEQ ID NO: 2.

Claims

1. A process for identifying a chemical compound which specifically binds to a human 5-HT_4B receptor, which comprises contacting nonneuronal cells expressing on their cell surface the human 5-HT_4B receptor having the amino acid sequence shown in FIG. 1 (SEQ ID NO. 2), with the chemical compound under conditions suitable for binding, and detecting specific binding of the chemical compound to the human 5-HT_4B receptor.

11. The process of any one of claims 1, 2, 3 4, 5, 6, 8, 9, 7, or 10, wherein the nonneuronal cell is a mammalian cell.

12. The process of claim 11, wherein the mammalian cell is a NIH 3T3 cell, a LM(tk-) cell, a COS 7 cell or a Chinese hamster ovary (CHO) cell.

2. A process for identifying a chemical compound which specifically binds to a human 5-HT_4B receptor, which comprises contacting a membrane fraction from a cell extract of nonneuronal cells expressing on their cell surface the human 5-HT_4B receptor having the amino acid sequence shown in FIG. 1 (SEQ ID NO. 2), with the chemical compound under conditions suitable for binding, and detecting specific binding of the chemical compound to the human 5-HT_4B receptor.

3. A process involving competitive binding for identifying a chemical compound which specifically binds to a human 5-HT_4B receptor, which comprises separately contacting nonneuronal cells expressing on their cell surface the human 5-HT_4B receptor having the amino acid sequence shown in FIG. 1 (SEQ ID NO. 2), with both the chemical compound and a second chemical compound known to bind to the human 5-HT_4B receptor, and with only the second chemical compound, under conditions suitable for binding of both compounds, and detecting specific binding of the chemical compound to the human 5-HT_4B receptor, a decrease in binding of the second chemical compound to the human 5-HT_4B in the presence of the chemical compound indicating that the chemical compound binds to the human 5-HT_4B receptor.

4. A process involving competitive binding for identifying a chemical compound which specifically binds to a human 5-HT_4B receptor, which comprises separately contacting a membrane fraction from a cell extract of nonneuronal cells expressing on their cell surface the human 5-HT_4B receptor having the amino acid sequence shown in FIG. 1 (SEQ ID NO. 2), with both the chemical compound and a second chemical compound known to bind to the human 5-HT_4B receptor, and with only the second chemical compound, under conditions suitable for binding of both compounds, and detecting specific binding of the chemical compound to the human 5-HT_4B receptor, a decrease in binding of the second chemical compound to the human 5-HT_4B in the presence of the chemical compound indicating that the chemical compound binds to the human 5-HT_4B receptor.

5. A process for determining whether a chemical compound specifically binds to and activates a human 5-HT_4B receptor, which comprises contacting nonneuronal cells producing a second messenger response and expressing on their cell surface the human 5-HT_4B receptor having the amino acid sequence shown in FIG. 1 (SEQ ID NO. 2), with the chemical compound under conditions suitable for activation of the human 5-HT_4B receptor, and measuring the second messenger response in the presence and in the absence of the chemical compound, a change in second messenger response in the presence of the chemical compound indicating that the chemical compound activates the human 5-HT_4B receptor.

7. The process of claim 5 or 6, wherein the second messenger response comprises adenylate cyclase activity and the change in second messenger response is an increase in adenylate cyclase activity.

6. A process for determining whether a chemical compound specifically binds to and activates a human 5-HT_4B receptor, which comprises contacting a membrane fraction from a cell extract of nonneuronal cells producing a second messenger response and expressing on their cell surface the human 5-HT_4B receptor having the amino acid sequence shown in FIG. 1 (SEQ ID NO. 2), with the chemical compound under conditions suitable for activation of the human 5-HT_4B receptor, and measuring the second messenger response in the presence and in the absence of the chemical compound, a change in second messenger response in the presence of the chemical compound indicating that the chemical compound activates the human 5-HT_4B receptor.

8. A process for determining whether a chemical compound specifically binds to and inhibits activation of a human 5-HT_4B receptor, which comprises separately contacting nonneuronal cells producing a second messenger response and expressing on their cell surface the human 5-HT_4B receptor having the amino acid sequence shown in FIG. 1 (SEQ ID NO. 2), with both the chemical compound and a second chemical compound known to activate the human 5-HT_4B receptor, and with only the second chemical compound, under conditions suitable for activation of the human 5-HT_4B receptor, and measuring the second messenger response in the presence of only the second chemical compound and in the presence of both the second chemical compound and the chemical compound, a smaller change in second messenger response in the presence of both the chemical compound and the second chemical compound than in the presence of only the second chemical compound indicating that the chemical compound inhibits activation of the human 5-HT_4B receptor.

10. The process of claim 8 or 9, wherein the second messenger response comprises adenylate cyclase activity and the change in second messenger response is a smaller increase in the level of adenylate cyclase activity in the presence of both the chemical compound and the second chemical compound than in the presence of only the second chemical compound.

9. A process for determining whether a chemical compound specifically binds to and inhibits activation of a human 5-HT_4B receptor, which comprises separately contacting a membrane fraction from a cell extract of nonneuronal cells producing a second messenger response and expressing on their cell surface the human 5-HT_4B receptor having the amino acid sequence shown in FIG. 1 (SEQ ID NO. 2), with both the chemical compound and a second chemical compound known to activate the human 5-HT_4B receptor, and with only the second chemical compound, under conditions suitable for activation of the human 5-HT_4B receptor, and measuring the second messenger response in the presence of only the second chemical compound and in the presence of both the second chemical compound and the chemical compound, a smaller change in second messenger response in the presence of both the chemical compound and the second chemical compound than in the presence of only the second chemical compound indicating that the chemical compound inhibits activation of the human 5-HT_4B receptor.