Compounds and methods of making same

• US 4,347,176 A
• Filed: 04/14/1980
• Issued: 08/31/1982
• Est. Priority Date: 04/14/1980
• Status: Expired due to Term

• Current Assignee: Burroughs Wellcome and Co.
• Original Assignee: Burroughs Wellcome and Co.
• Inventors: Mehta, Nariman B.
• Primary Examiner(s): Nucker, Christine M.
• Application Number:
  US06/140,157
• Time in Patent Office: 869 Days
• Field of Search: 424/1, 424/12, 435/7, 23/230 B, 560/39, 560/193, 422/61
• US Class Current: 530/363
• CPC Class Codes: A61K 47/646 the entire peptide or protein drug conjugate elicits an immune response, e.g. conjugate vaccines
• CPC Class Codes: A61K 51/0406 Amines, polyamines, e.g. spermine, spermidine, amino acids, (bis)guanidines
• CPC Class Codes: C07C 255/00 Carboxylic acid nitriles cyanogen or compounds thereof C01C3/00
• CPC Class Codes: C07C 45/00 Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
• CPC Class Codes: C07C 45/54 of compounds containing doubly bound oxygen atoms, e.g. esters
• CPC Class Codes: C07C 45/56 from heterocyclic compounds C07C45/55 takes precedence
• CPC Class Codes: C07C 49/82 containing hydroxy groups
• CPC Class Codes: C07C 49/825 all hydroxy groups bound to the ring
• CPC Class Codes: C07D 263/04 having no double bonds between ring members or between ring members and non-ring members
• CPC Class Codes: C07D 263/10 with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
• CPC Class Codes: Y10S 436/804 Radioisotope, e.g. radioimmunoassay
• CPC Class Codes: Y10S 436/822 Identified hapten
• CPC Class Codes: Y10S 530/806 Antigenic peptides or proteins

Abstract

A radioimmunoassay procedure has been discovered for bupropion [(±)-2-t-butylamino-3-chloropropiophenone], an antidepressant compound, in biological fluids. Novel compounds of formula ##STR1## wherein either R₂ is oxygen and R₁ is (CH₂)_n --0--(CO)_m --(CH₂)_p --COOH where n is an integer from 0 to 5, m is 0 or 1, and p is an integer from 1 to 4; or R₁ is hydrogen and R₂ is N--0--(CH₂)_q --COOH where q is an integer from 1 to 3, and novel methods of making them and novel intermediates useful therein are disclosed. Novel immunogens for raising bupropion specific antisera, and novel methods of making them are disclosed comprising conjugates of the novel compounds and suitable carrier material. The drug is added to the antisera together with novel radiolabeled competitor of formula ##STR2## wherein R'"'"' is a suitable radioisotope as described above and R₃ is hydrogen, R₄ is Cl, and R₅ is oxygen, or

R'"'"' is hydrogen and either

(a) R₃ is H and R₄ is Cl and R₅ is NO(CH₂)_q CO--X where q is as defined for formula I and X is a group having formula III ##STR3## wherein R₆ is a suitable radioisotope as described above, R₇ is hydroxyl, and s is an integer from 1 to 4;

(b) R₄ is Cl, R₅ is oxygen, and R₃ is (CH₂)_n --0--(CO)_m --(CH₂)_p --CO--X where n, m, and p are as defined for formula I and X is as defined above; or

(c) R₃ is hydroxy, R₅ is oxygen and R₄ is a suitable radioisotope as defined above.

The measure of radioactivity of the antisera bound radiolabeled competitor provides a measure of the amount of drug in the assayed biological fluid sample. A standard curve can be constructed from which the amount of drug in the assay sample can be interpolated. A kit is disclosed for practicing the radioimmunoassay.

Claims

1. A compound of the formula

wherein R₈ is (CH₂)_n --O--CO(CH₂)_p --CO₂ H

2. The compound according to claim 1 wherein R₈ is at the 4-position.

3. The compound according to claim 2 which is α

-t-butylamino-3-chloro-4-γ

-hydroxypropylpropiophenone

4. The method of making a compound of claim 1 comprising reacting an intermediate compound of formula ##STR35## wherein n is an defined in claim 1, with ##STR36## wherein p is as defined in claim 1.

5. A method according to claim 4 of making the compound of claim 3 comprising reacting intermediate compound ##STR37## with succinic anhydride.

8. A compound of the formula ##STR40## wherein R₉ is (CH₂)_n --O--CO(CH₂)pCO--X where n and p are as defined for claim 1 and X is ##STR41## wherein R₆ is a radioactive nuclide, R₇ is hydroxyl, and s is an integer from 1 to 4.

9. A compound according to claim 8 wherein said radioactive nuclide is selected from ³ H and ¹²⁵ I.

10. A compound according to claim 8 in which R₉ is γ

-hydroxyproplpropiophenone-hemisuccinate-¹²⁵ I-tyramine amide.

11. A compound according to claim 10 in which R₉ is at the 4-position.

12. A method of making a compound of claim 8 comprising reacting a compound of claim 1 with trialkylamino, the alkyl moiety of which has 1 to 4 carbons and alkyl haloformate, the alkyl moiety of which has 1 to 3 carbons, and subsequently adding the reaction mixture to

13. A method according to claim 12 wherein said trialkylamine is triethylamine, said alkyl halformate is isobutyl chloroformate and the reaction mixture is subsequently added to tyramine.

14. An immunogen comprising a conjugate comprising a compound of formula:

##STR42## wherein R₈ is as defined for claim 1, coupled to a suitable immunogenic carrier material.

15. An immunogen according to claim 14 wherein the immunogenic carrier material is bovine serum albumin.

16. An immunogen according to claim 15 in which R₈ is γ

-hydroxypropylpropiophenone-hemisuccinate and said carrier material is bovine serum albumin.

17. An immunogen according to claim 16 in which R₈ is at the 4-position.

18. A method of making an immunogen according to claim 14 comprising reacting said compound, said immunogenic carrier material and a suitable coupling agent.

6. A compound of the formula ##STR38## wherein n is an integer from 1 to 5.

7. The compound according to claim 6 which is ##STR39##