Activated amphetamines

• US 5,976,812 A
• Filed: 07/17/1997
• Issued: 11/02/1999
• Est. Priority Date: 07/25/1996
• Status: Expired due to Fees

• Current Assignee: Roche Diagnostics GmbH (Roche Holding AG)
• Original Assignee: Roche Diagnostics GmbH (Roche Holding AG)
• Inventors: Zink, Bruno, Vogel, Rudolf, Huber, Erasmus, Klein, Christian, Rollinger, Wolfgang
• Primary Examiner(s): MacMillan, Keith D.
• Assistant Examiner(s): WESSENDORF, TERESA D
• Application Number:
  US08/896,330
• Time in Patent Office: 838 Days
• Field of Search: 560/24, 560/145, 558/254, 435/721, 424/193.1, 424/178.1, 530/391.1, 530/391.7
• US Class Current: 435/7.1
• CPC Class Codes: C07D 207/416 with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to other ring carbon atoms
• CPC Class Codes: C07D 207/452 with hydrocarbon radicals, substituted by hetero atoms, directly attached to the ring nitrogen atom
• CPC Class Codes: G01N 33/946 CNS-stimulants, e.g. cocaine, amphetamines

Abstract

The invention concerns activated amphetamine derivatives, a process for their production as well as their use for producing immunogens or detection conjugates carrying amphetamine groups. A further subject matter of the invention are new conjugates carrying amphetamine groups, a process for the production thereof as well as their use for the production of antibodies or for the determination of amphetamine or derivatives thereof.

Claims

1. An activated amphetamine derivative having the structural formula ##STR3## in which R₁ is H or OH, R₂, R₃, R₄, and R₅ are independently of one another H, CH₃ or C₂ H₅, and L is a linker of formula (--(CH₂)_o --NH--C(O)--)_p --(CH₂)_q --X--, in which o is 2 to 6, p is 1 or 2, q is 2 to 10 and X is selected from the group consisting of O, NH and a bond, or a salt of said activated amphetamine derivative.

2. The activated amphetamine derivative of claim 1, wherein R₅ is H.

3. The activated amphetamine derivative of claim 1, wherein the linker is in the p-position relative to the ethylamine side chain.

4. The activated amphetamine derivative of claim 1, wherein p is 1 and X is a bond.

5. The activated amphetamine derivative of claim 1, wherein o is 2 and q is 4.

6. The activated amphetamine derivative of claim 1, wherein R₁, R₃ and R₅ are H, R₂ is CH₃ and R₄ is H or CH₃.

7. A conjugate, comprising at least one amphetamine group and a conjugation partner P, obtained by reacting the maleinimide group of an activated amphetamine derivative of claim 1 with an SH group of the conjugation partner, wherein said conjugate has the structural formula ##STR4##

8. A conjugate, having the structural formula in which P is a conjugation partner, R₁ to R₅ are defined as in claim 1, r is 0 to 10, s is 1 to 40 and L is defined as in claim 1, or a salt of said conjugate.

9. The conjugate of claim 8, wherein p is 1.

10. The conjugate of claim 8, wherein o is 2 and q is 4.

11. The conjugate of claim 8, wherein R₁, R₃ and R₅ are H, R₂ is CH₃ and R₄ is H or CH₃.

12. The conjugate of claim 8, wherein P is selected from the group consisting of a immunoglobulin, a enzyme, a dye, and a metal label.

13. The conjugate of claim 8, wherein said conjugate is bound to a solid phase or has a group which is capable of binding to a solid phase.

14. A process for the production of an activated amphetamine derivative of claim 1, wherein a linker is introduced on the aromatic ring of the amphetamine in a one-step or multiple-step synthesis wherein the amino group of the amphetamine side chain is protected during the synthesis, the derivative that is formed is reacted with maleinimide alkylamine, the protective group is removed and the activated amphetamine derivative is isolated.

15. A process for the production of a conjugate of claim 7, wherein an activated amphetamine derivative of claim 1 is reacted together with a reagent P--(--(CH₂)_r --SH)_s in which P, r and s are defined as claimed in claim 8 under such conditions that an addition of the SH groups to the double bond in the maleinimide functionality occurs and the reaction product is isolated.

16. A reagent kit comprising an activated amphetamine derivative of claim 1.

17. The conjugate of claim 12, wherein the dye is a fluorescent dye.

18. A reagent kit comprising the conjugate of claim 7.

19. A reagent kit comprising an activated amphetamine derivative characterized by the structural formula ##STR5## in which R₁ is H or OH, R₂, R₃, R₄ and R₅ are independently of one another H, CH₃ or C₂ H₅, and L is a linker as defined in claim 1 and a conjugate of claim 7.

20. A pharmaceutical composition comprising the conjugate of claim 7 and a pharmaceutically acceptable carrier.

21. A method for producing antibodies against an amphetamine or a derivative thereof comprising inoculating a subject with the pharmaceutical composition of claim 20.

22. A method for determining the presence of amphetamines or derivatives thereof in a sample comprising contacting said sample with the conjugate of claim 7 and determining presence of said amphetamines.