Antibodies to Olanzapine Haptens and Use Thereof

Abstract

Disclosed is an antibody which binds to olanzapine, which can be used to detect olanzapine in a sample such as in a competitive immunoassay method. The antibody can be used in a lateral flow assay device for point-of-care detection of olanzapine, including multiplex detection of aripiprazole, olanzapine, quetiapine, and risperidone in a single lateral flow assay device.

What is claimed is: 
     
         1 . An isolated antibody or a binding fragment thereof, which binds to olanzapine and which:
 (i) is generated in response to a conjugate of a compound of Formula I and an immunogenic carrier; or   (ii) competes for an epitope which is the same as an epitope bound by an antibody generated in response to a conjugate of a compound of Formula I and an immunogenic carrier,   
       
         
           
           
               
               
           
         
         wherein: 
         R 1  is H, 
       
       
         
           
           
               
               
           
         
       
       CH 2 NH 2 , CH 2 NHC(O)(CH 2 ) m CO 2 H, or Z—(Y) p -G;
 R 2  is H, 
 
       
         
           
           
               
               
           
         
       
       CH 2 NH 2 , CH 2 NHC(O)(CH 2 ) m CO 2 H, or Z—(Y) p -G;
 R 3  is H, or W—(Y) p -G; provided that two of R 1 , R 2 , R 3  must be H, and further provided that R 1 , R 2  and R 3  may not all be H simultaneously; 
 wherein: 
 Z is selected from the group consisting of: 
 —N(R 4 )—, —O—, —S—, -alkyl-, -alkoxyalkyl-, -aminoalkyl-, -thioalkyl-, -heteroalkyl-, alkylcarbonyl-, 
 
       
         
           
           
               
               
           
         
         wherein: 
         W is selected from the group consisting of: 
         —C(O)—, -alkyl-, -alkoxyalkyl-, -aminoalkyl-, -thioalkyl-, -heteroalkyl-, -alkylcarbonyl-, —N(R 4 )—, 
       
       
         
           
           
               
               
           
         
         R 4  is H, an alkyl group, cycloalkyl group, araalkyl group or substituted or unsubstituted aryl group; 
         Y is an organic spacer group; 
         G is a functional linking group capable of binding to a carrier; 
         p is 0, or 1; 
         m is 1, 2, 3, 4, or 5; 
         n is 1, 2, 3, 4, or 5. 
       
     
     
         2 . The antibody of  claim 1 , wherein the antibody is generated in response to a conjugate of a compound of Formula I and an immunogenic carrier. 
     
     
         3 . The antibody of  claim 1 , wherein the antibody fragment is selected from the group of fragments consisting of Fv, F(ab′), F(ab′)2, scFv, minibody and diabody fragments. 
     
     
         4 . The antibody of  claim 1 , wherein the antibody is a monoclonal antibody. 
     
     
         5 . An assay kit comprising the antibody of  claim 1 . 
     
     
         6 . An assay device comprising the antibody of  claim 1 . 
     
     
         7 . The assay device of  claim 6  wherein the device is a lateral flow assay device. 
     
     
         8 . A method of producing an antibody which binds to olanzapine, the method comprising:
 (i) selecting a host for antibody production; and   (ii) inoculating the host with a conjugate of a compound of Formula I and an immunogenic carrier, wherein the host produces an antibody which binds to olanzapine,   
       
         
           
           
               
               
           
         
         wherein: 
         R 1  is H, 
       
       
         
           
           
               
               
           
         
       
       CH 2 NH 2 , CH 2 NHC(O)(CH 2 ) m CO 2 H, or Z—(Y) p -G;
 R 2  is H, 
 
       
         
           
           
               
               
           
         
       
       CH 2 NH 2 , CH 2 NHC(O)(CH 2 ) m CO 2 H, or Z—(Y) p -G;
 R 3  is H, or W—(Y) p -G; provided that two of R 1 , R 2 , R 3  must be H, and further provided that R 1 , R 2  and R 3  may not all be H simultaneously; 
 wherein: 
 Z is selected from the group consisting of: 
 —N(R 4 )—, —O—, —S—, -alkyl-, -alkoxyalkyl-, -aminoalkyl-, -thioalkyl-, -heteroalkyl-, alkylcarbonyl-, 
 
       
         
           
           
               
               
           
         
         wherein: 
         W is selected from the group consisting of: 
         —C(O)—, -alkyl-, -alkoxyalkyl-, -aminoalkyl-, -thioalkyl-, -heteroalkyl-, -alkylcarbonyl-, —N(R 4 )—, 
       
       
         
           
           
               
               
           
         
         R 4  is H, an alkyl group, cycloalkyl group, araalkyl group or substituted or unsubstituted aryl group; 
         Y is an organic spacer group; 
         G is a functional linking group capable of binding to a carrier; 
         p is 0, or 1; 
         m is 1, 2, 3, 4, or 5; 
         n is 1, 2, 3, 4, or 5. 
       
     
     
         9 . A method of producing a hybridoma cell line capable of producing a monoclonal antibody which binds to olanzapine, the method comprising:
 (i) selecting a host for antibody production;   (ii) inoculating the host with a conjugate of a compound of Formula I and an immunogenic carrier;   (iii) fusing a cell line from said inoculated host with a continuously dividing cell to create a fused cell capable of producing a monoclonal antibody which binds to olanzapine; and   (iv) cloning the fused cell so as to obtain a hybridoma cell line,   
       
         
           
           
               
               
           
         
         wherein: 
         R 1  is H, 
       
       
         
           
           
               
               
           
         
       
       CH 2 NH 2 , CH 2 NHC(O)(CH 2 ) m CO 2 H, or Z—(Y) p -G;
 R 2  is H, 
 
       
         
           
           
               
               
           
         
       
       CH 2 NH 2 , CH 2 NHC(O)(CH 2 ) m CO 2 H, or Z—(Y) p -G;
 R 3  is H, or W—(Y) p -G; provided that two of R 1 , R 2 , R 3  must be H, and further provided that R 1 , R 2  and R 3  may not all be H simultaneously; 
 wherein: 
 Z is selected from the group consisting of: 
 —N(R 4 )—, —O—, —S—, -alkyl-, -alkoxyalkyl-, -aminoalkyl-, -thioalkyl-, -heteroalkyl-, alkylcarbonyl-, 
 
       
         
           
           
               
               
           
         
         wherein: 
         W is selected from the group consisting of: 
         —C(O)—, -alkyl-, -alkoxyalkyl-, -aminoalkyl-, -thioalkyl-, -heteroalkyl-, -alkylcarbonyl-, —N(R 4 )—, 
       
       
         
           
           
               
               
           
         
         R 4  is H, an alkyl group, cycloalkyl group, araalkyl group or substituted or unsubstituted aryl group; 
         Y is an organic spacer group; 
         G is a functional linking group capable of binding to a carrier; 
         p is 0, or 1; 
         m is 1, 2, 3, 4, or 5; 
         n is 1, 2, 3, 4, or 5. 
       
     
     
         10 . A method of detecting olanzapine in a sample, the method comprising:
 (i) contacting a sample with an antibody of  claim 1  labeled with a detectable marker, wherein the labeled antibody and olanzapine present in the sample form a labeled complex; and   (ii) detecting the labeled complex so as to detect olanzapine in the sample.   
     
     
         11 . A competitive immunoassay method for detecting olanzapine in a sample, the method comprising:
 (i) contacting a sample with the antibody of  claim 1 , and with olanzapine or a competitive binding partner of olanzapine, wherein one of the antibody and the olanzapine or competitive binding partner thereof is labeled with a detectable marker, and wherein sample olanzapine competes with the olanzapine or competitive binding partner thereof for binding to the antibody; and   (ii) detecting the label so as to detect sample olanzapine.   
     
     
         12 . The method of  claim 11  wherein the olanzapine or competitive binding partner thereof is labeled with the detectable marker. 
     
     
         13 . The method of  claim 11  wherein the antibody is labeled with a detectable marker. 
     
     
         14 . The method of  claim 11  wherein the immunoassay is performed on a lateral flow assay device and the sample is applied to the device. 
     
     
         15 . The method of  claim 10  or  11 , further comprising detecting the presence of one or more analytes in addition to olanzapine. 
     
     
         16 . The method of  claim 15  wherein the one or more analytes are anti-psychotic drugs other than olanzapine. 
     
     
         17 . The method of  claim 16  wherein the anti-psychotic drugs other than olanzapine are selected from the group consisting of: risperidone, paliperidone, quetiapine, aripiprazole, and metabolites thereof. 
     
     
         18 . The method of  claim 10  or  11 , wherein the detection of olanzapine is an indication of patient adherence with prescribed olanzapine therapy. 
     
     
         19 . The method of  claim 10  or  11 , wherein the detection of olanzapine is used to determine whether a patient should be converted from an oral olanzapine regimen to an injectable anti-psychotic regimen. 
     
     
         20 . The method of  claim 10  or  11 , wherein the detection of olanzapine is used to determine if the dose level or dosing interval of oral or injectable olanzapine should be increased or decreased to ensure attainment or maintenance of efficacious or safe drug levels. 
     
     
         21 . The method of  claim 10  or  11 , wherein the detection of olanzapine is an aid in the initiation of olanzapine therapy by providing evidence of the attainment of minimum pK levels. 
     
     
         22 . The method of  claim 10  or  11 , wherein the detection of olanzapine is used to determine bioequivalence of olanzapine in multiple formulations or from multiple sources. 
     
     
         23 . The method of  claim 10  or  11 , wherein the detection of olanzapine is used to assess the impact of polypharmacy and potential drug-drug interactions. 
     
     
         24 . The method of  claim 10  or  11 , wherein the detection of olanzapine is an indication that a patient should be excluded from or included into a clinical trial and is an aid in the subsequent monitoring of adherence to clinical trial medication requirements.