Compounds for the treatment of sexual dysfunction

• US 6,991,916 B2
• Filed: 09/07/2001
• Issued: 01/31/2006
• Est. Priority Date: 07/14/2000
• Status: Expired due to Fees

• Current Assignee: Pfizer Inc.
• Original Assignee: Pfizer Inc.
• Inventors: Wayman, Christopher Peter, Phillips, Stephen Charles, Singleton, David Harlan, Boyd, Helen Frances, Benson, Neil, Contillo, Leonard Gabriel, Walsh, Roderick Thomas, Harrow, Ian Dennis, Stacey, Peter
• Primary Examiner(s): Prouty, Rebecca E.
• Assistant Examiner(s): Ramirez, Delia M.
• Application Number:
  US09/948,429
• Publication Number:
  US 20020177689A1
• Time in Patent Office: 1,607 Days
• Field of Search: 435/24, 435/212, 435/226, 436/501, 436/86, 530/350
• US Class Current: 435/24
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/00 Medicinal preparations containing organic active ingredients
• CPC Class Codes: A61K 31/165 having aromatic rings, e.g. colchicine, atenolol, progabide
• CPC Class Codes: A61K 31/17 having the group >N—C(O)—N< or >N—C(S)—N<, e.g. urea, thiourea, carmustine isoureas, isothioureas A61K31/155; sulfonylureas A61K31/64
• CPC Class Codes: A61K 31/18 Sulfonamides compounds containing a para-N-benzene-sulfonyl-N- group A61K31/63
• CPC Class Codes: A61K 31/196 the amino group being directly attached to a ring, e.g. anthranilic acid, mefenamic acid, diclofenac, chlorambucil
• CPC Class Codes: A61K 31/395 having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
• CPC Class Codes: A61K 31/4015 having oxo groups directly attached to the heterocyclic ring, e.g. piracetam, ethosuximide
• CPC Class Codes: A61K 31/433 Thidiazoles
• CPC Class Codes: A61K 31/4412 having oxo groups directly attached to the heterocyclic ring
• CPC Class Codes: A61K 31/454 containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
• 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: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 15/10 for impotence
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C09B 11/24 Phthaleins containing amino groups ; Phthalanes; Fluoranes; Phthalides; Rhodamine dyes; Phthaleins having heterocyclic aryl rings; Lactone or lactame forms of triarylmethane dyes
• CPC Class Codes: C12N 9/6421 from mammals
• CPC Class Codes: C12N 9/6489 Metalloendopeptidases (3.4.24)
• CPC Class Codes: C12Q 1/37 : involving peptidase or proteinase
• CPC Class Codes: G01N 2500/00 : Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 2800/344 : Disorders of the penis and the scrotum and erectile dysfuncrion
• CPC Class Codes: G01N 33/542 : with steric inhibition or signal modification, e.g. fluorescent quenching

Abstract

Polynucleotide and polypeptide sequences are described. The polypeptide sequences comprise one or more of: (a) a polypeptide having the deduced amino acid sequence translated from the polynucleotide sequence in SEQ ID NO: 1 or SEQ ID NO: 5 and variants, fragments, homologues, analogues and derivatives thereof; (b) a polypeptide of SEQ ID NO: 2 and variants, fragments, homologues, analogues and derivatives thereof; (c) a polypeptide encoded by the cDNA of NCIMB 41110 and variants, fragments, homologues, analogues and derivatives thereof; or (d) a polypeptide which has at least 78% identity to (i) the polypeptide encoded by the polynucleotide of SEQ ID NO: 1 or SEQ ID NO: 5, (ii) the polypeptide of SEQ ID NO: 2, or (iii) the polypeptide encoded by the cDNA of NCIMB 41110. Such polypeptide sequences are, inter alia, useful in the prophylaxis and/or treatment of sexual dysfunction, in particular male erectile dysfunction (MED) or female sexual dysfunction (FSD), preferably female sexual arousal disorder (FSAD).

Claims

1. A method for identifying an agent which binds to and/or modulates a soluble secreted endopeptidase (SEP) polypeptide comprising contacting said agent with a SEP polypeptide comprising:

the amino acid sequence of SEQ ID NO;

2, an amino acid sequence at least 95% identical to SEQ ID NO;

2, an amino acid sequence encoded by the polynucleotide sequence of SEQ ID NO;

1 or SEQ ID NO;

5;

or a polypeptide encoded by the cDNA of NCIMB 41110, and determining whether binding and/or modulation occurs.

2. The method of claim 1, comprising contacting said SEP polypeptide with a SEP substrate peptide in the presence of said agent, wherein said substrate peptide is capable of providing a detectable signal in response to cleavage by said SEP polypeptide, wherein said agent is identified as a modulator of said SEP polypeptide if there is a difference in the detectable signal in the presence and in the absence of said agent.

3. The method of claim 2, wherein said method identifies an agent that decreases said detectable signal and is a SEP polypeptide inhibitor.

4. The method of claim 3, wherein said substrate peptide is labelled with at least one fluorescent donor dye and said signal is detected by Fluorescence Resonance Energy Transfer (FRET) assay.

5. The method of claim 4, wherein said labelled substrate peptide is Rhodamine green-Gly-Gly-dPhe-Leu-Arg-Arg-Val-Cys(QSY™

7)-β

Ala-NH₂.

6. The method of claim 4, wherein said labelled substrate peptide is 5-(and 6) tetramethyl rhodamine Gly-Gly-dPhe-Leu-Arg-Arg-Val-Cys(QSY™

7)-β

Ala-NH₂.

7. The method of claim 4, wherein said labelled substrate peptide is 5-carboxyfluorescein-Gly-Gly-dPhe-Leu-Arg-Arg-Val-Cys(5-(and 6)tetramethylrhodamine)-β

Ala-NH₂.

8. The method of claim 1, wherein binding between said SEP polypeptide and said agent is detected.

9. The method of claim 8, wherein said method is a competitive binding assay.

10. A method for identifying an agent which inhibits SEP or neutral endopeptidase (NEP), wherein said SEP has at least 95% identity to the human SEP amino acid sequence of SEQ ID NO:

2, and wherein said NEP comprises the human NEP amino acid sequence of SEQ ID NO;

63, comprising contacting said SEP or NEP and a peptidase substrate peptide selected from the group consisting of Rhodamine green-Gly-Gly-dPhe-Leu-Arg-Arg-Val-Cys(QSY™

7)-β

Ala-NH₂, 5-(and

     6) tetramethyl rhodamine Gly-Gly-dPhe-Leu-Arg-Arg-Val-Cys(QSY™

7)-β

Ala-NH₂, and 5-carboxyfluorescein-Gly-Gly-dPhe-Leu-Arg-Arg-Val-Cys (5-(and

     6) tetramethylrhodamine)-β

Ala-NH₂ in the presence of said agent, wherein said substrate peptide is capable of providing a detectable signal in response to cleavage by said SEP or NEP as detected by Fluorescence Resonance Energy Transfer (FRET) assay, wherein said agent is identified as an inhibitor of said SEP or NEP if there is a decrease in the detectable signal in the presence of said agent as compared to in the absence of said agent.