Selective Targeting of Apoptotic Cells

Abstract

The invention relates to a method of selectively targeting an active agent (or agent capable of becoming an active agent) to apoptotic cells in a vertebrate, comprising administering to said vertebrate a system comprising an arsenoxide (or arsenoxide equivalent) compound and said agent, wherein said system selectively targets apoptotic cells.

1 - 101 . (canceled) 
     
     
         102 . A method of selectively targeting an active agent (or agent capable of becoming an active agent) to apoptotic cells, comprising contacting said cells with a system comprising an arsenoxide (or arsenoxide equivalent) compound and said agent, wherein said arsenoxide (or arsenoxide equivalent) compound selectively targets said active agent (or agent capable of becoming an active agent) to apoptotic cells. 
     
     
         103 . The method according to  claim 102  wherein the system comprises an arsenoxide (or arsenoxide equivalent) compound cleavably linked to an active agent or an agent capable of becoming an active agent. 
     
     
         104 . The method according to  claim 102 , wherein the system comprises:
 a first component comprising an arsenoxide (or arsenoxide equivalent) compound inked to a first binding member; and   a second component, comprising a second binding member capable of binding to said first binding member, wherein said second binding member is an active agent or an agent capable of becoming an active agent.   
     
     
         105 . The method according to  claim 104 , wherein the first binding member is an enzyme and the second binding member is a substrate for the enzyme. 
     
     
         106 . The method according to  claim 105 , wherein the substrate for the enzyme is a pro-agent which is converted to an active agent by the enzyme. 
     
     
         107 . The method according to  claim 106 , wherein the pro-agent is a prodrug which is converted to an active drug by the enzyme. 
     
     
         108 . The method according to  claim 102 , wherein the system comprises:
 a first component comprising an arsenoxide (or arsenoxide equivalent) compound linked to a first binding member; and   a second component comprising a second binding member capable of binding to said first binding member linked to at least one active agent (or agent capable of becoming an active agent).   
     
     
         109 . The method according to  claim 108 , wherein the first binding member is biotin and the second binding member is selected from avidin and streptavidin. 
     
     
         110 . The method according to  claim 108 , wherein the first binding member is selected from avidin and streptavidin, and the second binding member is biotin. 
     
     
         111 . The method according to  claim 108 , wherein the first binding member is methotrexate and the second binding member is dihydrofolate reductase (DHFR). 
     
     
         112 . The method according to  claim 108 , wherein the first binding member is dihydrofolate reductase and the second binding member is methotrexate. 
     
     
         113 . The method according to  claim 108 , wherein the first binding member is hirudin and the second binding member is thrombin. 
     
     
         114 . The method according to  claim 108 , wherein the first binding member is thrombin and the second binding member is hirudin. 
     
     
         115 . The method according to  claim 108 , wherein the first binding member is an antigen and the second binding member is an antibody. 
     
     
         116 . The method according to  claim 108 , wherein the second binding member is directly or indirectly linked to the active agent. 
     
     
         117 . The method according to  claim 116 , wherein said indirect linking between the active agent and the second binding member is by virtue of a further binding interaction between the second binding member and at least one further binding member, wherein the further binding member is directly linked to the active agent. 
     
     
         118 . The method according to  claim 108 , wherein the first binding member is biotin, the second binding member is selected from avidin and streptavidin, and the second binding member is indirectly linked to an active agent by virtue of a further binding interaction between the avidin or streptavidin and at least one further biotin moiety which is directly linked to the active agent. 
     
     
         119 . The method according to  claim 102 , wherein the system comprises:
 a first component comprising an arsenoxide (or arsenoxide equivalent) compound linked to a first binding member;   a second component comprising a second binding member capable of binding to said first binding member linked to an enzyme; and   a substrate for said enzyme.   
     
     
         120 . The method according to  claim 119 , wherein the substrate for said enzyme is a pro-agent which is converted to an active agent by the enzyme. 
     
     
         121 . The method according to  claim 120 , wherein the pro-agent is a prodrug. 
     
     
         122 . The method according to  claim 102 , wherein the active agent is a therapeutic agent or a diagnostic agent. 
     
     
         123 . The method according to  claim 122 , wherein the therapeutic agent is selected from the group consisting of radionucleotides, chemotherapeutic agents, cytotoxins, coagulants, growth factors, cytokines, bacterial, plant and fungal endotoxins. 
     
     
         124 . The method according to  claim 123 , wherein the radionucleotides are selected from the group consisting of:  3 H,  11 C,  14 C,  15 O,  13 N,  32 P,  33 P,  35 S,  18 F,  125 I,  127 I,  111 In,  105 Rh,  153 Sm,  67 Cu,  67 Ga,  166 Ho,  177 Lu,  186 Re,  188 Re, and  99m Tc. 
     
     
         125 . The method according to  claim 123 , wherein the chemotherapeutic agents are selected from the group consisting of adriamycin, taxol, fluorouricil, melphalan, cisplatin, alpha interferon, vincristine, vinblastine, angioinhibins, TNP-470, pentosan polysulfate, platelet factor 4, angiostatin, LM-609, SU-101, CM-101, Techgalan, thalidomide, SP-PG, alkylating agents, nitrosoureas, alkyl sulfonates triazines, ethylenimines, folic acid analogues, pyrimidine analogues, cytosine arabinoside, purine analogues, antitumour antibiotics, anthracyclines, hormones and hormone antagonists, brequinar, and regimens including COMP (cyclophosphamide, vincristine, methotrexate and prednisone), etoposide, mBACOD (methotrexate, bleomycin, doxorubicin, cyclophosphamide, vincristine and dexamethasone), and PROMACE/MOPP (prednisone, methotrexate (w/leucovin rescue), doxorubicin, cyclophosphamide, taxol, etoposide/mechlorethamine, vincristine, prednisone and procarbazine). 
     
     
         126 . The method according to  claim 123 , wherein the cytotoxin is ricin. 
     
     
         127 . The method according to  claim 123 , wherein the bacterial, plant and fungal endotoxins are selected from the group consisting of: ribosome inactivating protein, diphtheria toxin, pseudomonas endotoxin, A chain toxin, α-sarcin, aspergillin, restrictocin, and ribonucleases. 
     
     
         128 . The method according to  claim 122  wherein the diagnostic agent is an imaging agent. 
     
     
         129 . The method according to  claim 122  wherein the diagnostic agent is selected from the group consisting of fluorescent labels, radionucleotides, paramagnetic ions, X-ray imaging agents, chemiluminescent labels or labels which are detectable through a secondary enzymatic or binding step. 
     
     
         130 . The method according to  claim 129 , wherein the fluorescent label is selected from Cy™5.5, and fluorescein. 
     
     
         131 . The method according to  claim 129  wherein the radionucleotide is selected from the group consisting of  3 H,  11 C,  14 C,  15 O,  13 N,  32 P,  33 P,  35 S,  18 F,  125 I,  127 I,  127 I,  111 In,  105 Rh,  153 Sm,  67 Cu,  67 Ga,  166 Ho,  177 Lu,  186 Re,  188 Re, and  99m Tc. 
     
     
         132 . The method according to  claim 129 , wherein the paramagnetic ions are selected from the group consisting of: chromium(III), gadolinium(III), iron(II), iron (III), holmium(III), erbium(III), manganese(II), nickel(II), copper(II), neodymium(III), yttrium(III), samarium(III), and dysprosium(III). 
     
     
         133 . The method according to  claim 132 , wherein the paramagnetic ion is gadolinium(III). 
     
     
         134 . The method according to  claim 129 , wherein the X-ray imaging agents are selected from the group consisting of: gold(III), lead(II), lanthanum(III) and bismuth(III). 
     
     
         135 . The method according to  claim 122 , wherein the active agent resides within a vehicle for delivery of said agent. 
     
     
         136 . The method according to  claim 135 , wherein the vehicle for the active agent is a liposome. 
     
     
         137 . The method according to  claim 102 , wherein the arsenoxide (or arsenoxide equivalent) compound is of the formula (I):
   A—[(XBX′) n B′—Y] p    (I)   
       wherein
 A comprises at least one pendant group; 
 (XBX′) n B′ comprises a suitable linker group, wherein 
 X is selected from the group consisting of —NR—, —S(O)—, —S(O)O—, —S(O) 2 —, —S(O) 2 O—, —C(O)—, —C(S)—, —C(O)O—, —C(S)O—, —C(S)S—, —P(O)(R 1 )—, and —P(O)(R 1 )O—, or is absent; 
 B is selected from the group consisting of C 1 -C 10  alkylene, C 2 -C 10  alkenylene, C 2 -C 10  alkynylene, C 3 -C 10  cycloalkylene, C 5 -C 10  cycloalkenylene, C 3 -C 10  heterocycloalkylene, C 5 -C 10  heterocycloalkenylene, C 6 -C 12  arylene, heteroarylene, and C 2 -C 10  acyl; 
 X′ is selected from the group consisting of —NR—, —O—, —S—, —Se—, —S—S—, —S(O)—, —OS(O)—, —OS(O)O—, —OS(O) 2 —, —OS(O) 2 O—, —S(O)O—, —S(O) 2 —, —S(O) 2 —, —OP(O)(R 1 )—, —OP(O)(R 1 )O—, —OP(O)(R 1 )OP(O)(R 1 )O—, —C(O)—, —C(S)—, —C(O)O—, C(S)O—, —C(S)S—, —P(O)(R 1 )—, —P(O)(R 1 )O—, and 
 
       
         
           
           
               
               
           
         
       
       or is absent; wherein E is O, S, Se, NR or N(R) 2   + ;
 B′ is selected from the group consisting of C 1 -C 10  alkylene, C 2 -C 10  alkenylene, C 2 -C 10  alkynylene, C 3 -C 10  cycloalkylene, C 5 -C 10  cycloalkenylene, C 3 -C 10  heterocycloalkylene, C 5 -C 10  heterocycloalkenylene, C 6 -C 12  arylene, and heteroarylene, or is absent; and wherein 
 each R is independently selected from the group consisting of hydrogen, C 1 -C 10  alkyl, C 2 -C 10  alkenyl, C 2 -C 10  alkynyl, C 3 -C 10  cycloalkyl, C 5 -C 10  cycloalkenyl, C 3 -C 10  heterocycloalkyl, C 5 -C 10  heterocycloalkenyl, C 6 -C 12  aryl, heteroaryl, OR 2 , and C 2 -C 10  acyl; 
 R′ is the same as R, or two R′ may be taken together with the nitrogen atoms to which they are attached to form a 5 or 6-membered saturated or unsaturated heterocyclic ring; 
 each R 1  is independently selected from the group consisting of hydrogen, C 1 -C 10  alkyl, C 2 -C 10  alkenyl, C 2 -C 10  alkynyl, C 3 -C 10  cycloalkyl, C 5 -C 10  cycloalkenyl, C 3 -C 10  heterocycloalkyl, C 5 -C 10  heterocycloalkenyl, C 6 -C 12  aryl, heteroaryl, halo, OR 2  and N(R) 2 ; 
 each R 2  is independently selected from the group consisting of hydrogen, C 1 -C 10  alkyl, C 2 -C 10  alkenyl, C 2 -C 10  alkynyl, C 3 -C 10  cycloalkyl, C 5 -C 10  cycloalkenyl, C 3 -C 10  heterocycloalkyl, C 5 -C 10  heterocycloalkenyl, C 6 -C 12  aryl, heteroaryl and —C(O)R 5 ; 
 each R 5  is independently selected from the group consisting of hydrogen, C 1 -C 10  alkyl, C 2 -C 10  alkenyl, C 2 -C 10  alkynyl, C 3 -C 10  cycloalkyl, C 5 -C 10  cycloalkenyl, C 3 -C 10  heterocycloalkyl, C 5 -C 10  heterocycloalkenyl, C 6 -C 12  aryl, heteroaryl, C 1 -C 10  alkoxy, C 3 -C 10  alkenyloxy, C 3 -C 10  alkynyloxy, C 3 -C 10  cycloalkyloxy, C 5 -C 10  cycloalkenyloxy, C 3 -C 10  heterocycloalkyloxy, C 5 -C 10  heterocycloalkenyloxy, C 6 -C 12  aryloxy, heteroaryloxy, C 1 -C 10  alkylthio, C 3 -C 10  alkenylthio, C 3 -C 10  alkynylthio, C 3 -C 10  cycloalkylthio, C 5 -C 10  cycloalkenylthio, C 3 -C 10  heterocycloalkylthio, C 5 -C 10  heterocycloalkenylthio, C 6 -C 12  arylthio, heteroarylthio, OH, SH and N(R) 2 ; 
 wherein for each instance that B and/or B′ is arylene, the substituents directly attached to the respective arylene rings (including arsenoxide or arsenoxide equivalent) may be in a para-, meta- or ortho- relationship; and 
 wherein each alkylene, alkenylene, alkynylene, cycloalkylene, cycloalkenylene, heterocycloalkylene, heterocycloalkenylene, arylene, heteroarylene, and acyl may be independently substituted with hydrogen, C 1 -C 10  alkyl, C 2 -C 10  alkenyl, C 2 -C 10  alkynyl, C 3 -C 10  cycloalkyl, C 5 -C 10  cycloalkenyl, C 3 -C 10  heterocycloalkyl, C 5 -C 1 O heterocycloalkenyl, C 6 -C 12  aryl, heteroaryl, cyano, cyanate, isocyanate, OR 2a , SR 6 , nitro, arsenoxide, —S(O)R 3 , —OS(O)R 3 , —S(O) 2 R 3 , —OS(O) 2 R 3 , —P(O)R 4 R 4 , —OP(O)R 4 R 4 , —N(R″) 2 , —NRC(O)(CH 2 ) m Q, —C(O)R 5 ; 
 
       
         
           
           
               
               
           
         
         wherein R, R 1  and R 5  are as defined above; and 
         R 2a  is selected from the group consisting of hydrogen, C 1 -C 5  alkyl, C 2 -C 5  alkenyl, C 2 -C 5  alkynyl, C 3 -C 10  cycloalkyl, C 5 -C 10  cycloalkenyl, C 6 -C 12  aryl, —S(O)R 3 , —S(O) 2 R 3 , —P(O)(R 4 ) 2 , —N(R) 2 , and —C(O)R 5 ; 
         each R 3  is independently selected from the group consisting of hydrogen, C 1 -C 10  alkyl, C 2 -C 10  alkenyl, C 2 -C 10  alkynyl, C 3 -C 10  cycloalkyl, C 5 -C 10  cycloalkenyl, C 3 -C 10  heterocycloalkyl, C 5 -C 10  heterocycloalkenyl, C 6 -C 12  aryl, heteroaryl, C 1 -C 10  alkoxy, C 3 -C 10  alkenyloxy, C 3 -C 10  alkynyloxy, C 3 -C 10  cycloalkyloxy, C 5 -C 10  cycloalkenyloxy, C 3 -C 10  heterocycloalkyloxy, C 5 -C 10  heterocycloalkenyloxy, C 6 -C 12  aryloxy, heteroaryloxy, C 1 -C 10  alkylthio, C 3 -C 10  alkenylthio, C 3 -C 10  alkynylthio, C 3 -C 10  cycloalkylthio, C 5 -C 10  cycloalkenylthio, C 3 -C 10  heterocycloalkylthio, C 5 -C 10  heterocycloalkenylthio, C 6 -C 12  arylthio, heteroarylthio, and N(R) 2 ; 
         each R 4  is independently selected from the group consisting of hydrogen, C 1 -C 10  alkyl, C 2 -C 10  alkenyl, C 2 -C 10  alkynyl, C 3 -C 10  cycloalkyl, C 5 -C 10  cycloalkenyl, C 3 -C 10  heterocycloalkyl, C 5 -C 10  heterocycloalkenyl, C 6 -C 12  aryl, heteroaryl, C 1 -C 10  alkoxy, C 3 -C 10  alkenyloxy, C 3 -C 10  alkynyloxy, C 3 -C 10  cycloalkyloxy, C 5 -C 10  cycloalkenyloxy, C 3 -C 10  heterocycloalkyloxy, C 5 -C 10  heterocycloalkenyloxy, C 6 -C 12  aryloxy, heteroaryloxy, C 1 -C 10  alkylthio, C 3 -C 10  alkenylthio, C 3 -C 10  alkynylthio, C 3 -C 10  cycloalkylthio, C 5 -C 10  cycloalkenylthio, C 3 -C 1 O heterocycloalkylthio, C 5 -C 10  heterocycloalkenylthio, C 6 -C 12  arylthio, heteroarylthio, halo, and N(R) 2 ; 
         R 6  is selected from the group consisting of C 1 -C 10 alkyl, C 2 -C 10  alkenyl, C 2 -C 10  alkynyl, C 3 -C 10  cycloalkyl, C 5 -C 10  cycloalkenyl, C 3 -C 10  heterocycloalkyl, C 5 -C 10  heterocycloalkenyl, C 6 -C 12  aryl, heteroaryl, C 1 -C 10  alkylthio, C 3 -C 10  alkenylthio, C 3 -C 10  alkynylthio, C 3 -C 10  cycloalkylthio, C 5 -C 10  cycloalkenylthio, C 3 -C 10  heterocycloalkylthio, C 5 -C 10  heterocycloalkenylthio, C 6 -C 12  arylthio, heteroarylthio, —S(O)R 3 , —S(O) 2 R 3 , and —C(O)R 5 , 
         R″ is the same as R, or two R″ taken together with the N atom to which they are attached may form a saturated, unsaturated or aromatic heterocyclic ring system; 
         Q is selected from halogen and —OS(O) 2 Q 1 ; wherein Q 1  is selected from C 1 -C 4  alkyl, C 1 -C 4  perfluoroalkyl, phenyl, and p-methylphenyl; and 
         m is 1 to 5; 
         n is an integer from 0 to 20; 
         Y comprises at least one arsenoxide or arsenoxide equivalent; and 
         p is an integer from 1 to 10. 
       
     
     
         138 . The method according to  claim 137 , wherein the sum total of carbon atoms in A and L together, is greater than 6. 
     
     
         139 . The method according to  claim 137 , wherein A is selected from the group consisting of natural, unnatural and synthetic amino acids, hydrophilic amines, peptides, polypeptides, oligosaccharides, and thiol containing proteins, or a combination thereof. 
     
     
         140 . The method according to  claim 137 , wherein A is selected from the group consisting of glutathione, glucosamine, cysteinylglycine, cysteic acid, aspartic acid, glutamic acid, lysine, and arginine, and wherein the sulfur atom of each sulfur containing compound is optionally oxidised to form a sulfoxide or sulfone. 
     
     
         141 . The method according to  claim 140 , wherein A is glutathione. 
     
     
         142 . The method according to  claim 137 , wherein p is an integer from 1 to 5. 
     
     
         143 . The method according to  claim 137 , wherein in the formula (XBX′) n B′, n is an integer selected from the group consisting of from 0 to 15, 0 to 10, 0 to 5, and 0 to 1. 
     
     
         144 . The method of  claim 143 , wherein n is 0, 1, or 2. 
     
     
         145 . The method according to  claim 144 , wherein the compound is 4-(N-(S-glutathionylacetyl)amino)-phenylarsenoxide, (GSAO), according to Formula IV: 
       
         
           
           
               
               
           
         
       
     
     
         146 . The method according to  claim 102 , wherein said method comprises the steps of:
 (a) administering a first component of the system;   (b) optionally waiting for a period of time; and   (c) administering a second component of the system.   
     
     
         147 . The method according to  claim 102 , wherein said method comprises the steps of:
 (a) administering a first component of the system;   (b) optionally waiting for a period of time;   (c) administering a second component of said system;   (d) optionally waiting for a further period of time; and   (e) administering a further binding member linked to an active agent (or agent capable of becoming an active agent).   
     
     
         148 . The method according to  claim 146  or  147 , wherein the period of time is selected from the group consisting of between about 1 hour and about 48 hours; between about 3 and about 36 hours, between about 6 hours and about 24 hours, and about 18 hours.