Fluorescent somatostatin

• US 5,824,772 A
• Filed: 06/07/1995
• Issued: 10/20/1998
• Est. Priority Date: 04/04/1995
• Status: Expired due to Fees

• Current Assignee: Advanced Bioconcept Company
• Original Assignee: Advanced Bioconcept, Inc. (Revvity, Inc.)
• Inventors: Gaudriault, Georges, Vincent, Jean-Pierre, Beaudet, Alain
• Primary Examiner(s): Tsang, Cecilia J.
• Assistant Examiner(s): DELACROIX MUIRHEI, CYBILLE
• Application Number:
  US08/475,751
• Time in Patent Office: 1,231 Days
• Field of Search: 530/311, 435/7.1
• US Class Current: 530/311
• CPC Class Codes: C07K 14/575 Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin C07K14/665, e.g. corticotropin C07K14/695
• CPC Class Codes: C07K 14/57536 Endothelin, vasoactive intestinal contractor [VIC]
• CPC Class Codes: C07K 14/57563 Vasoactive intestinal peptide [VIP]; Related peptides
• CPC Class Codes: C07K 7/14 Angiotensins: Related peptides

Abstract

Light-emitting compounds of the formula: ##STR1## where R₁ is a light-emitting moiety and R₂ is a somatostatin-based peptide featuring a first amino acid sequence -Phe-Phe-Trp-Lys-Thr- or (SEQ ID NO:1) or -Phe-Phe-D-Trp-Lys-Thr-. The peptide is linked at a first amino acid position to (C-X), and the light-emitting compound exhibits substantial biological activity in the presence of a receptor having affinity for somatostatin-based peptides.

Claims

1. A compound of the formula:

##STR3## wherein R₁ is a light-emitting moiety;

R₂ is Y-Z-Q, wherein

Z comprises -Phe-Phe-Trp-Lys-Thr- (SEQ ID NO;

1) or -Phe-Phe-D-Trp-Lys-Thr-;

each of Y and Q is an amino acid sequence such that the number of amino acids in R₂ is between 7 and 28; and

CX is C═

O, C═

S, --CHOH, C═

C═

O, C═

NH, -CH₂, --CHOR, --C═

NR, --CHR, --CR₃ R₄, wherein each of R, R₄, and R₃, independently, is H or a C₁ -C₆ branched or unbranched, substituted or unsubstituted alkyl; and

C of X-C is bonded via an --NH-- or --S-- group to an amino acid of R₂ ; and

wherein said compound, when compared to R₂ alone, retains at least 25% of its binding affinity for the human somatostatin receptor in vivo.

2. The compound of claim 1, wherein R₂ is a somatostatin-based peptide.

3. The compound of claim 2, wherein -Phe-Phe-Trp-Lys-Thr- (SEQ ID NO:

1) or -Phe-Phe-D-Trp-Lys-Thr- is located at positions 7-11 in said somatostatin-based peptide.

4. The compound of claim 2, wherein said somatostatin-based peptide further comprises -Ala-Gly-Cys-Lys-Asn-Phe- (SEQ ID NO:

2).

5. The compound of claim 4, wherein -Ala-Gly-Cys-Lys-Asn-Phe- (SEQ ID NO:

2) is comprised by the N-terminus of said somatostatin-based peptide.

6. The compound of claim 5, wherein said -Ala- residue of said somatostatin-based peptide is attached via said amino or thiol group NH or --S-- group to (C-X).

7. The compound of claim 2, wherein said somatostatin-based peptide further comprises -Phe-Thr-Ser-Cys- (SEQ ID NO:

3).

8. The compound of claim 7, wherein -Phe-Thr-Ser-Cys- (SEQ ID NO:

3) is comprised by the C-terminus of said somatostatin-based peptide.

9. The compound of claim 2, wherein said somatostatin-based peptide further comprises -Ala-Gly-Cys-Lys-Asn-Phe- (SEQ ID NO:

2) and -Phe-Thr-Ser-Cys- (SEQ ID NO;

     3).

10. The compound of claim 9, wherein said somatostatin-based peptide is somatostatin-14 or D-Trp⁸ -somatostatin-14.

11. The compound of claim 1, wherein said light-emitting moiety is selected from the group consisting of Bodipy, fluorescein, FTC, Texas red, phycoerythrin, rhodamine, carboxytetra-methylrhodamine, DAPI, indopyras dyes, Cascade blue, coumarins, NBD, Lucifer Yellow, propidium iodide, and derivatives thereof.

12. The compound of claim 1, wherein C-X is C═

O or C═

S ═

O or ═

S.

13. The compound of claim 1, wherein said compound is a pharmaceutically acceptable salt or complex thereof.

14. A method for labelling a receptor having an affinity for a somatostatin-based peptide by contacting said receptor with the compound of claim 1.

15. A compound of claim 1, wherein the number of amino acids in R₂ is between 7 and 14.

21. A compound of claim 15, wherein the number of amino acids in R₂ is between 7 and 14.

16. A compound of the formula:

##STR4## wherein R₁ is a light-emitting moiety;

R₂ is Y-Z-Q, wherein Z comprises -Phe-Phe-Trp-Lys-Thr- (SEQ ID NO;

1) or -Phe-Phe-D-Trp-Lys-Thr-;

each of Y and Q is an amino acid sequence such that the number of amino acids in R₂ is between 7 and 28; and

CX is C═

O, C═

S, --CHOH, C═

C═

O, C═

NH, --CH₂, --CHOR, --C═

NR, --CHR, --CR₃ R₄, wherein each of R, R₄, and R₃, independently, is H or a C₁ -C₆ branched or unbranched, substituted or unsubstituted alkyl; and

C of X-C is bonded via an --NH-- or --S-- group to an amino acid of R₂ ; and

wherein said compound is selected from a mixture including said compound and secondary compounds having biological activities less than 0.25% of the biological activity of R₂ --H in the presence of a receptor having affinity for somatostatin-based peptides.

17. The compound of claim 16, wherein said selection is accomplished by exposing said compound to a receptor recognizing a somatostatin-based peptide, and then selectively isolating said exposed compound if said compound exhibits substantial biological activity.

18. The compound of claim 17, wherein said compound having substantial biological activity is selected using high-pressure liquid chromatography.

19. The compound of claim 16, wherein R₂ is a somatostatin-based peptide.

20. The compound of claim 19, wherein said -Phe-Phe-Trp-Lys-Thr- (SEQ ID NO:

1) or -Phe-Phe-D-Trp-Lys-Thr- is located at positions 7-11 of said somatostatin-based peptide.