Novel oxazine-ureas and thiazine urea chromophors as fluorescent labels

• US 4,714,763 A
• Filed: 07/11/1985
• Issued: 12/22/1987
• Est. Priority Date: 07/11/1985
• Status: Expired due to Fees

• Current Assignee: Viomedics Incorporated
• Original Assignee: Viomedics Incorporated
• Inventors: Theodoropulos, Spyros
• Primary Examiner(s): Raymond, Richard L.
• Application Number:
  US06/753,937
• Time in Patent Office: 894 Days
• Field of Search: 544/31.37, 544/101, 544/103
• US Class Current: 544/31
• CPC Class Codes: C07D 265/38 [b, e]-condensed with two six-membered rings
• CPC Class Codes: C07D 279/36 [b, e]-condensed, at least one with a further condensed benzene ring
• CPC Class Codes: C07J 43/003 not condensed
• CPC Class Codes: C09B 19/00 Oxazine dyes
• CPC Class Codes: C09B 21/00 Thiazine dyes
• CPC Class Codes: G01N 33/533 with fluorescent label

Abstract

Novel urea derivatives of oxazine and thiazine chromophors have the structural formula I or II. ##STR1## wherein M is oxygen or sulfur, R₁ and R₂ are aliphatic alkyl groups or hydrogen; R₃ is hydrogen or alkyl group; R₄ is hydrogen, alkyl or amine group; R₅ is hydrogen, amine or alkyl group; X.sup.⊖ is an anion consisting of an organic (e.g. CH₃ COO.sup.⊖, CH₃ CH₂ COO.sup.⊖ and the like) or inorganic specie (e.g. Cl.sup.⊖, Br.sup.⊖, I.sup.⊖, ClO₄.sup.⊖, SO₄ ", and the like); n is 0 to 20; Z is N═C═O, N═C═S, carboxylic, primary or secondary amine, and when n=0, Z may be ##STR2## wherein Q is hydroxyl, amino, carboxylic, sulfydryl, isocyanato, or isothiocyanato.

The functional oxazine-urea and thiazine-urea derivatives react with compounds of interest to form adducts resulting in the fluorescent labeling of the compound.

Claims

1. Urea derivatives of oxazine and thiazine chromophors selected from the group consisting of:

##STR21## wherein n is 0 to 20;

M is oxygen or sulfur;

R₁ is hydrogen or alkyl group from 1 to 10 carbon atoms;

R₂ is hydrogen or alkyl group from 1 to 10 carbon atoms;

R₃ is hydrogen or alkyl group from 1 to 10 carbon atoms;

R₄ is hydrogen, alkyl group from 1 to 10 carbon atoms, halogen or amino group;

R₅ is hydrogen, alkyl group from 1 to 10 carbon atoms, halogen or amino group;

X.sup.⊖

is an organic or an inorganic anion and Z is N═

C═

O, N═

C═

S, carboxylic, or amino, and when n=0, Z may be ##STR22## wherein Q is isocyanato or isothiocyanato.

2. Urea derivatives of oxazine and thiazine chromophors having the structural formula ##STR23## wherein n is 0 to 20;

M is oxygen or sulfur;

R₁ is hydrogen or alkyl group from 1 to 10 carbon atoms;

R₂ is hydrogen or alkyl group from 1 to 10 carbon atoms;

R₃ is hydrogen or alkyl group from 1 to 10 carbon atoms;

R₄ is hydrogen, alkyl group from 1 to 10 carbon atoms, halogen or amino group;

R₅ is hydrogen, alkyl group from 1 to 10 carbon atoms, halogen or amino group;

X.sup.⊖

is an anion consisting of an organic specie CH₃ COO.sup.⊖

, CH₃ CH₂ COO.sup.⊖

, or CF₃ COO.sup.⊖

, or an inorganic specie Cl.sup.⊖

, Br.sup.⊖

, I.sup.⊖

, ClO₄.sup.⊖

or SO₄^-2, Z is N═

C═

O, N═

C═

S, carboxylic, or amino, and when n=0, Z may be ##STR24## wherein Q is isocyanato or isothiocyanato.

7. The urea derivative of claim 2 which is isothiocyanato pentyl toluidine blue O urea.

8. The urea derivative of claim 2 which is isothiocyanato pentyl Brilliant cresyl blue urea.

3. Urea derivatives of oxazine and thiazine chormophors having the structural formula ##STR25## wherein n is 0 to 20;

M is oxygen or sulfur;

R₁ is hydrogen or alkyl group from 1 to 10 carbon atoms;

R₂ is hydrogen or alkyl group from 1 to 10 carbon atoms;

R₃ is hydrogen or alkyl group from 1 to 10 carbon atoms;

R₄ is hydrogen, alkyl group from 1 to 10 carbon atoms, halogen or amino group;

Z is N═

C═

O, N═

C═

S, carboxylic, or amino and when n=0, Z may be ##STR26## wherein Q is isocyanato or isothiocyanato, X.sup.⊖

is an anion consisting of an organic specie CH₃ COO.sup.⊖

, CH₃ CH₂ COO.sup.⊖

, CF₃ COO.sup.⊖

or an inorganic specie Cl.sup.⊖

, Br.sup.⊖

, I.sup.⊖

, ClO₄.sup.⊖

, SO₄^-2.

4. The urea derivative of claim 3 which is isothiocyanato pentyl Nile Blue A urea.

5. The urea derivative of claim 3 which is isocyanatohexyl Nile Blue A urea.

6. The urea derivative of claim 3 which is N-(2-thiolactone)-cresyl violet urea.