Method and kit for detecting, or determining 3,4- methylenedioxymethamphetamine

Abstract

The invention provides a hapten, an immunogen comprising the hapten coupled to an antigenicity-conferring carrier material, a conjugate comprising the hapten coupled to a labelling agent, as well as, antibodies raised against the aforementioned immunogen and capable of binding with MDMA.

1 - 29 . (canceled) 
   
   
       30 . A compound of the formula: 
     
       
         
         
             
             
         
       
       wherein: 
       R 19  is lower alkyl or is taken together with R 20  to form a ring, which may be a five- or six-member ring, usually a five-member ring; 
       R 20  is lower alkyl, or is taken together with R 19  to form a ring as discussed above, 
       R 1  is H or lower alkyl, 
       R 2  is
 (a) —(CH 2 ) a  C(O)(CH 2 ) b  SR 3 , wherein a is 0 to 5, b is 1 to 5 and R 3  is lower alkyl or (CH 2 ) c  C(O)NR 4 R 5  wherein c is 1 to 5, R 4  is H or lower alkyl and R 5  is H, an immunogenic carrier or a label, or 
 (b) (A) d (Q) n  wherein Q is H or —(CH 2 ) e  CH(R 8 )(CH 2 ) f  OC(O)(CH 2 ) g  R 9  being H only when d is 1 wherein A is —C(O)(CH 2 ) h  C(O)NR 10 ((CH 2 ) j  O(CH 2 ) k  O) m  (CH) 2 NR 11 —, d is 0 or 1, n is 0 or 1 wherein one of d or n is 1, h is 1 to 5, R 10  is H or lower alkyl, j is 1 to 5, k is 1 to 5, m is 1 to 3, R 11  is H or lower alkyl, e is 1 to 5, R 8  is OH or H, f is 1 to 5, g is 0 to 5, and R 9  is H, an immunogenic carrier or a label; 
 
       W is H or JR 14  being H when R 2  is other than H or lower alkyl, wherein 
       J is O or S, 
       R 14  is H, lower alkyl, a protecting group, or —(CH 2 ) r  C(O)NR 15  (CH 2 ) s  (D) t  R 16 , wherein r is 1 to 5, R 15  is H or lower alkyl, s is 1 to 5, D is S, O, or NH, t is 0 or 1 being 0 when R 16  is maleimidyl or succinimidyl, R 16  is H, maleimidyl, succinimidyl or —(CH 2 ) q  C(O)NR 17 R 18 , 
       q is 1 to 5, 
       R 17  is H or lower alkyl, 
       R 18  is H, lower alkyl, an immunogenic carrier or label, 
       and including the acid salts thereof. 
     
   
   
       31 . A compound according to  claim 30 , wherein R 1  is H or lower alkyl, W is H and R 2  is —(CH 2 ) a  C(O)(CH 2 ) b  SR 3 , wherein R 3  is —(CH 2 ) c  C(O)NR 4 R 5  wherein R 4  is H or lower alkyl and R 5  is an immunogenic carrier. 
   
   
       32 . A compound of the formula: 
     
       
         
         
             
             
         
       
       wherein: 
       R 1 ′ is H, lower alkyl or a protecting group, 
       R 2 ′ is 
       (a) —(CH 2 ) a  C(O)(CH 2 ) b  SR 3 ′, wherein a is 0 to 5, b is 1 to 5 and R 3 ′ is lower alkyl or (CH 2 ) c  C(O)NR 4 ′ R 5 ′ wherein c is 1 to 5, R 4 ′ is H or lower alkyl and R 5 ′ is H, an immunogenic carrier or a label, or 
       (b) (A) d (Q) n  wherein Q is H or —(CH 2 ) e  (CH(R 8 ′)(CH 2 ) f  OC(O)(CH 2 ) g  R 9 ′ being H only when d is 1 wherein A is —C(O)(CH 2 ) h  C(O)N(R 10 )((CH 2 ) j O(CH 2 ) k O) m  (CH) 2  NR 11 —, d is 0 or 1, n is 0 or 1 wherein one of d or n is 1, h is 1 to 5, R 10  is H or lower alkyl, j is 1 to 5, k is 1 to 5, m is 1 to 3, R 11  is H or lower alkyl, e is 1 to 5, R 8 ′ is OH or H, f is 1 to 5, g is 0 to 5, and R 9 ′ is H, an immunogenic carrier or a label, 
       and including the acid salts thereof. 
     
   
   
       33 . A compound according to  claim 32 , wherein R 1 ′ is H or lower alkyl and R 2 ′ is —(CH 2 ) a  C(O)(CH 2 ) b  SR 3 ′ wherein a is 0, b is 1, R 3 ′ is (CH 2 ) c  C(O)NR 4 ′R 5 ′ wherein c is 1, R 4 ′ is H and R 5 ′ is an immunogenic carrier. 
   
   
       34 . A compound according to  claim 32 , wherein R 1 ′ is H or lower alkyl and R 2 ′ is (A) d (Q) n  wherein d is 0, n is 1, Q is —(CH 2 ) e  CH(R 8 ′)(CH 2 ) f  OC(O)(CH 2 ) g  R 9 ′, e is 1, R 8 ′ is OH, f is 1, g is 0, and R 9 ′ is a poly(amino) acid. 
   
   
       35 . A compound according to  claim 34 , wherein said poly(amino) acid is an enzyme or an immunogenic carrier. 
   
   
       36 . A compound according to  claim 35 , wherein R 1 ′ is H or lower alkyl and R 2 ′ is (A) d (Q) n  wherein d is 1, n is 1, Q is —(CH 2 ) e CH(R 8 ′)(CH 2 ) f  OC(O)(CH 2 ) g  R 9 ′, e is 1, R 8 ′ is OH, f is 1, g is 0, A is —C(O)(CH 2 ) h  C(O)NR 10  ((CH 2 ) j  O(CH 2 ) k O) m (CH) 2 NR 11 —, R 10  is H, h is 2, m is 1, j is 2, R 10 ′ is H and R 9 ′ is a poly(amino) acid or a particle label or a particle immunogenic carrier. 
   
   
       37 . A compound according to  claim 36 , wherein R 9 ′ is a poly(amino) acid, which is an enzyme or an immunogenic carrier. 
   
   
       38 . An antibody raised against a compound according to  claim 35 , wherein said poly(amino) acid is an immunogenic carrier. 
   
   
       39 . An antibody raised against a compound according to  claim 37 , wherein said poly(amino) acid is an immunogenic carrier. 
   
   
       40 . A reagent system comprising a compound according to  claim 35 , wherein said poly(amino) acid is an enzyme, an antibody for methylenedioxyamphetamine and/or an antibody for methylenedioxymethamphetamine and/or an antibody for methylenedioxyethamphetamine. 
   
   
       41 . A reagent system comprising a compound according to  claim 37 , wherein said poly(amino) acid is an enzyme, an antibody for methylenedioxyamphetamine and/or an antibody for methylenedioxymethamphetamine and/or an antibody for methylenedioxyethamphetamine. 
   
   
       42 . A method for determining methylenedioxyamphetamine and/or methylenedioxymethamphetamine and/or methylenedioxyethamphetamine in a sample suspected of containing methylenedioxyamphetamine and/or methylenedioxymethamphetamine and/or methylenedioxyethamphetamine, said method comprising:
 (a) providing in combination in a medium:
 (i) said sample and 
 (ii) a reagent system according to claim  11 ; and 
   (b) examining said medium for the presence of a complex comprising said methylenedioxyamphetamine and said antibody for methylenedioxyamphetamine and/or a complex of said methylenedioxymethamphetamine and said antibody for methylenedioxymethamphetamine, the presence thereof indicating the presence of said methylenedioxyamphetamine and/or methylenedioxymethamphetamine and/or methylenedioxyethamphetamine in said sample.   
   
   
       43 . A method for determining methylenedioxyamphetamine and/or methylenedioxymethamphetamine and/or methylenedioxyethamphetamine in a sample suspected of containing methylenedioxyamphetamine and/or methylenedioxymethamphetamine and/or methylenedioxyethamphetamine, said method comprising:
 (a) providing in combination in a medium:
 (i) said sample and 
 (ii) a reagent system according to claim  12 ; and 
   (c) examining said medium for the presence of a complex comprising said methylenedioxyamphetamine and said antibody for methylenedioxyamphetamine and/or a complex of said methylenedioxymethamphetamine and said antibody for methylenedioxymethamphetamine and/or a complex of said methylenedioxyethamphetamine and said antibody for methylenedioxyethamphetamine, the presence thereof indicating the presence of said methylenedioxyamphetamine and/or methylenedioxymethamphetamine and/or methylenedioxyethamphetamine in said sample.   
   
   
       44 . A compound of the formula: 
     
       
         
         
             
             
         
       
     
     wherein:
 R 1  is H, lower alkyl, a protecting group, or is taken together with R 2  to form a ring, 
 R 2  is H, lower alkyl, a protecting group, —(CH 2 ) n  SCH 2 C(O)R 6  or —(CH 2 ) n  C(SO 2 R 6 )═CH 2 , or is taken together with R 1  to form a ring, 
 R 3  and R 4  are independently H or lower alkyl or a protecting group, or, when R 1  is taken together with R 2  to form a ring, at least one of R 3  or R 4  is —C(O)(CH 2 ) n  R 5 , —C(O)(CH 2 ) n  NHC(O)R 5 , —C(O)(CH 2 ) n  NHC(O)(CH 2 ) n  SR 5 , —(CH 2 ) n  C(SO 2 R 5 )═CH 2 , —(CH 2 ) n  SCH 2 C(O)R 5 , or —(CH 2 ) n  C(SO 2 R 5 )═CH 2 , or when R 1  is not taken together with R 2  to form a ring, R 2  is not H or lower alkyl or a protecting group, 
 R 5  is H, —OH, —SH, —O-lower alkyl, halogen, NH 2 , -succinimidyl, -maleimidyl, protein, or label, 
 R 6  is H, —OH, —SH, —O-lower alkyl, halogen, NH 2 , -succinimidyl, -maleimidyl, protein, or label, and 
 n is an integer from 1 to 5, 
 
     and including acid salts thereof. 
   
   
       45 . Antibodies raised against the compound of  claim 44 . 
   
   
       46 . A compound of the formula: 
     
       
         
         
             
             
         
       
       wherein:
 R 7  is H, lower alkyl, a protecting group, —C(O)(CH 2 ) n R 5 , —C(O)(CH 2 ) n  NHC(O)R 5 , —C(O)(CH 2 ) n  NHC(O)(CH 2 ) n SR 5 , —(CH 2 ) n C(SO 2 R 5 )═CH 2 , —(CH 2 ) n SCH 2 C(O)R 5 , or —(CH 2 ) n  C(SO 2 R 5 )═CH 2 , 
 
       R 8  is H, lower alkyl, a protecting group, —C(O)(CH 2 ) n  R 5 , —C(O)(CH 2 ) n NHC(O)R 5 , —C(O)(CH 2 ) n  NHC(O)(CH 2 ) n SR 5 —(CH 2 ) n  C(SO 2 R 5 )═CH 2 , —(CH 2 ) n SCH 2 C(O)R 5 , or —(CH 2 ) n  C(SO 2 R 5 )═CH 2 , 
       R 5  is H, —OH, —SH, —O-lower alkyl, halogen, NH 2 , immunogenic carrier, -succinimidyl, -maleimidyl, or label, and 
       n is an integer from 1 to 5, 
       with the proviso that at least one of R 7  and R 8  are not H or lower alkyl, and including the acid salts thereof. 
     
   
   
       47 . A compound according to  claim 46 , wherein said immunogenic carrier is a protein selected from the group consisting of KLH, BSA, BGG, and ovalbumin. 
   
   
       48 . Antibodies raised against the compound of  claim 47 . 
   
   
       49 . A method for determining a compound selected from the group consisting of 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxy-methamphetamine (MDMA), 3,4-methylenedioxyethylamphetamine (MDEA) and 4-hydroxy-3-methoxy-methamphetamine (HMMA), said method comprising:
 (a) providing in combination in a medium:
 (i) a sample suspected of containing said compound and 
 (ii) an antibody raised against a compound of the formula: 
   
     
       
         
         
             
             
         
       
       wherein: 
       R 1  is H, lower alkyl, a protecting group, or is taken together with R 2  to form a ring, 
       R 2  is H, lower alkyl, a protecting group, —(CH 2 ) n  SCH 2 C(O)R 6  or —(CH 2 ) n  C(SO 2 R 6 )═CH 2 , or is taken together with R 1  to form a ring, 
       R 3  and R 4  are independently H or lower alkyl or a protecting group, or, when R 1  is taken together with R 2  to form a ring, at least one of R 3  or R 4  is —C(O)(CH 2 ) n R 5 , —C(O)(CH 2 ) n  NHC(O)R 5 , —C(O)(CH 2 ) n  NHC(O)(CH 2 ) n SR 5 , —(CH 2 ) n  C(SO 2 R 5 )═CH 2 , —(CH 2 ) n  SCH 2 C(O)R 5 , or —(CH 2 ) n  C(SO 2 R 5 )═CH 2 , or when R1 is not taken together with R 2  to form a ring, R 2  is not H or lower alkyl or a protecting group, 
       R 5  is an immunogenic carrier, 
       R 6  is an immunogenic carrier, and 
       n is an integer from 1 to 5, and 
       (b) examining said medium for the presence of a complex comprising said compound and said antibody, the presence thereof indicating the presence of said compound in said sample. 
     
   
   
       50 . A method according to  claim 49 , wherein said protein is selected from the group consisting of KLH, BSA, BGG and ovalbumin.