Process for preparing targeted forms of methyltrithio antitumor agents
AssigneeAmerican Cyanamid Company
InventorsWilliam James McGahren, Martin Leon Sassiver, George A. Ellestad, Philip R. Hamann, Lois M. Hinman, Janis Upeslacis
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Abstract
This disclosure describes a method for constructing targeting agent drug conjugates from the family of methyltrithio antibacterial and antitumor agents.
Claims21
A process for preparing the targeted derivatives ##STR22## of compounds of formula CH.sub.3 SSS-W, wherein CH.sub.3 SSS-W is an N-acyl derivative of an antitumor antibiotic LL-E33288 .alpha..sub.a.sup.Br, .alpha..sub.2.sup.I, .beta..sub.1.sup.Br, .beta..sub.1.sup.I, .gamma..sub.1.sup.Br, .gamma..sub.1.sup.I, .delta..sub.1.sup.I, BBM-1675, FR-900405, FR-900406, PD 114759, PD 115028, CL-1577A, CL-1577B, CL-1577D, CL-1577E or CL-1724 wherein W is ##STR23## each of R.sub.4 and R.sub.7 independently is H or ##STR24## where R.sub.5 is CH.sub.3, C.sub.2 H.sub.5, or (CH.sub.3).sub.2 CH;
- X is an iodine or bromine atom;
- R.sup.5 is the group RCO wherein R is hydrogen or a branched or unbranched alkyl (C.sub.1 -C.sub.10) or alkylene (C.sub.1 -C.sub.10) group, an aryl or heteroarylgorup, or an aryl-alkyl (C.sub.1 -C.sub.5) or heteroaryl-alkyl (C.sub.1 -C.sub.5) group, all optionally substituted by one or more hydroxy, amino, carboxy, halo, nitro, lower (C.sub.1 -C.sub.3) alkoxy, or lower (C.sub.1 -C.sub.5) thioalkoxy groups, comprising
- reacting CH.sub.3 SSS-W with a compound of formula Q-Sp-SH, wherein Sp is a straight or branched-chain divalent or trivalent (C.sub.1 -C.sub.18) radical, divalent or trivalent (C.sub.6 -C.sub.11) aryl or heteroaryl radical, divalent or trivalent (C.sub.3 -C.sub.18) cycloalkyl or heterocycloalkyl radical, divalent or trivalent (C.sub.6 -C.sub.11) aryl- or heteroaryl-alkyl (C.sub.4 -C.sub.18) radical, divalent or trivalent (C.sub.3 -C.sub.18) cycloalkyl or heterocycloalkyl-alkyl (C.sub.4 -C.sub.18) radical, divalent or trivalent (C.sub.2 -C.sub.18) unsaturated alkyl radical, wherein heteroaryl is (4-methyl-coumarin-7yl)amino and wherein if Sp is a trivalent radical, it can be additionally substituted by amino, (C.sub.1 -C.sub.10) alkylamino, (C.sub.6 -C.sub.11) arylamino, heteroarylamino, carboxyl, lower alkoxy, hydroxy, thiol, or lower alkylthio groups;
- and Q is halogen, amino, C.sub.1 -C.sub.10 alkylamino, carboxyl, carboxaldehyde, hydroxy or lower alkyldicarboxyl anhydride in acetonitrile in the presence of one equivalent of triethylamine or one equivalent of triethylamine and one equivalent of acetic acid at -10.degree. to -30.degree. C. for 1-48 hours,
- isolating the intermediate of formula Q-Sp-SS-W, wherein Q, Sp, and W are as hereinbefore defined, then reacting the compound of formula Q-Sp-SS-W, wherein Sp and W are as hereinbefore defined and Q is halogen, amino, alkylamino, carboxyl, carboxaldehyde, hydroxy, or lower alkyldicarboxylic anhydride with a molecule of the formula Tu-(Y).sub.n, wherein Tu-(Y).sub.n is a mono- or polyclonal antibody, its tumor-associated antigen-binding fragments, its chemically or genetically manipulated tumor-associated antigen-binding counterparts, or growth factors or steroids;
- Y is a side-chain amino or carboxy functionality of the antibody and n is 1-100, in aqueous buffer at a pH of between 6.5 and 9, at 40.degree. to 40.degree. C. either directly or in the presence of a water-soluble carbodiimide, to generate the compound ##STR25## wherein Tu, Sp, W, n, and Y are as hereinbefore defined, m is 1-15 and Z is formed from covalent reaction of the groups Q and Y and is --CONH--, --NH--, ##STR26## --N.dbd.CH--, or --CO.sub.2 --.
A process for preparing the targeted derivatives of compounds of formula CH.sub.3 SSS-W, wherein CH.sub.3 SSS-W is an N-acyl derivative of an antitumor antibiotic LL-E33288 .alpha..sub.2.sup.Br, .alpha..sub.2.sup.I, .beta..sub.1.sup.Br, .beta..sub.1.sup.I, .gamma..sub.1.sup.Br, .gamma..sub.1.sup.I, .delta..sub.1.sup.I, BBM-1675, FR-900405, FR-900406, PD 114759, PD 115028, CL-1577A, CL-1577B, CL-1577D, CL-1577E or CL-1724 wherein W is ##STR37## each of R.sub.6 and R.sub.7 independently is H or ##STR38## where R.sub.5 is CH.sub.3, C.sub.2 H.sub.5, or (CH.sub.3).sub.2 CH;
- X is an iodine or bromine atom;
- R.sub.5 is the group RCO wherein R is hydrogen or a branched or unbranched alkyl (C.sub.1 -C.sub.10) or alkylene (C.sub.1 -C.sub.10) group, an aryl or heteroaryl group, or an aryl-alkyl (C.sub.1 -C.sub.5) or heteroaryl-alkyl (C.sub.1 -C.sub.5) group, all optionally substituted by one or more hydroxy, amino, carboxy, halo, nitro, lower (C.sub.1 -C.sub.3, alkoxy, or lower (C.sub.1 -C.sub.5) thioalkoxy groups, comprising
- reacting the compound of formula Q-Sp-SS-W, wherein Sp is a straight or branched-chain divalent or trivalent (C.sub.1 -C.sub.18) radical, divalent or trivalent (C.sub.6 -C.sub.11) aryl or heteroaryl radical, divalent or trivalent (C.sub.3 -C.sub.18) cycloalkyl or heterocycloalkyl radical, divalent or trivalent (C.sub.6 -C.sub.11) aryl- or heteroaryl-alkyl (C.sub.4 -C.sub.18) radical, divalent or trivalent (C.sub.3 -C.sub.18) cycloalkyl or heterocycloalkyl-alkyl (C.sub.4 -C.sub.18) radical, divalent or trivalent (C.sub.2 -C.sub.18) unsaturated alkyl radical, wherein heteroaryl is (4-methyl-coumarin-7-yl)amino and wherein if Sp is a trivalent radical, it can be additionally substituted by amino, (C.sub.1 -C.sub.10) alkylamino, (C.sub.6 -C.sub.11) arylamino, heteroarylamino, carboxyl, lower alkoxy, hydroxy, thiol, or lower alkylthio groups;
- and Q is --NH.sub.2, --CONHNH.sub.2, --NHCONHNH.sub.2, --NHCSNHNH.sub.2, or --ONH.sub.2 with a molecule of formula Tu-(Y).sub.n wherein Tu is a mono- or polyclonal antibody, its tumor-associated antigen-binding fragments, its chemically or genetically manipulated tumor-associated antigen-binding counterparts, or growth factors or steroids;
- Y is an aldehyde generated from carbohydrate residues on Tu by oxidation in the presence of an alkaline earth periodate, in an aqueous buffer at a pH between 4.0 and 6.5 at 4.degree. to 40.degree. C., inclusive, and n is 1 to 20 to generate a compound of formula:
- ##STR39## wherein Tu, Sp, W, Y, and n are as hereinbefore defined and Z is formed from the covalent reaction of Q and Y and is --ON.dbd.CH--, --N.dbd.CH--, --CONHN.dbd.CH--, --NHCONHN.dbd.CH--, or --NHCSNHN.dbd.CH--, and m is 0.1 to 15.
A process for preparing the targeted derivatives ##STR40## of compounds of formula CH.sub.3 SSS-W, wherein CH.sub.3 SSS-W is an N-acyl derivative of an antitumor antibiotic LL-E33288 .alpha..sub.2.sup.Br, .alpha..sub.2.sup.I, .beta..sub.1.sup.Br, .gamma..sub.1.sup.I, .gamma..sub.1.sup.Br, .gamma..sub.1 BBM-1675, FR-900405, FR-900406, PD 114759, PD 115028, CL-1577A, CL-1577B, CL-1577D, CL-1577E or CL-1724 wherein W is ##STR41## each of R.sub.6 and R.sub.7 independently is H or ##STR42## where R.sub.5 is CH.sub.3, C.sub.2 H.sub.5, or (CH.sub.3).sub.2 CH;
- X is an iodine or bromine atom;
- R.sub.5' is the group RCO wherein R is hydrogen or a branched or unbranched alkyl (C.sub.1 -C.sub.10) or alkylene (C.sub.1 -C.sub.10) group an aryl or heteroaryl group, or an aryl-alkyl (C.sub.1 -C.sub.5) or heteroaryl-alkyl (C.sub.1 -C.sub.5) group, all optionally substituted by one or more hydroxy, amino, carboxy halo, nitro, lower (C.sub.1 -C.sub.3) alkoxy, or lower (C.sub.1 -C.sub.5) thioalkoxy groups, comprising
- reacting the compound of formula Q-Sp-SS-W, wherein Sp is a straight or branched-chain divalent or trivalent (C.sub.1 -C.sub.5) radical, divalent or trivalent (C.sub.6 -C.sub.11) aryl or heteroaryl radical, divalent or trivalent (C.sub.3 -C.sub.18) cycloalkyl or heterocycloalkyl radical, divalent or trivalent (C.sub.6 -C.sub.11) aryl or heteroaryl-alkyl (C.sub.2 -C.sub.18) radical, divalent or trivalent (C.sub.3 -C.sub.18) cycloalky heterocycloalkyl-alkyl (C.sub.4 -C.sub.18) radical, divalent or trivalent (C.sub.2 -C.sub.18) unsaturated alkyl radical, wherein heteroaryl is (4-methyl-coumarin-7-yl)amino and wherein if Sp is a trivalent radical, it can be additionally substituted by amino, (C.sub.1 -C.sub.10) alkylamino, (C.sub.6 -C.sub.11) arylamino, heteroarylamino, carboxyl, lower alkoxy, hydroxy, thiol, or lower alkylthio groups;
- and Q is --NH.sub.2, --CONHNH.sub.2, --NHCONHNH.sub.2, --NHCSNHNH.sub.2, or --ONH.sub.2 with a molecule of formula Tu-(Y).sub.n wherein Tu is a mono- or polyclonal antibody, its tumor-associated antigen-binding fragments, its chemically or genetically manipulated tumor-associated antigen-binding counterparts, or growth factors or steroids;
- Y is an aldehyde generated from carbohydrate residues of the antibody by oxidation in the presence of an alkaline earth periodate, in an aqueous buffer at a pH between 4.0 and 6.5, at 4.degree. to 40.degree. C., inclusive, and n is 1 to 20 to generate a compound of formula:
- ##STR43## wherein Tu, Sp, W, Y, and n are as hereinbefore defined and Z is formed form the covalent reaction of Q and Y and is --ON.dbd.CH--, --N.dbd.CH--, --CONHN.dbd.CH--, or --NHCSNHN.dbd.CH--, and m is 0.1 to 15;
- and treating the compound immediately hereinabove of formula:
- ##STR44## wherein Tu, Z, Sp, W, Y, n, and m are as immediately hereinabove defined with acteylhydrazine or tyrosine hydrazine in an aqueous buffer at a pH between 4.0 and 6.5, at 4.degree. to 40.degree. C., inclusive, to generate a compound of formula:
- ##STR45## wherein Tu, Z, Sp, W, n, and m are as immediately hereinabove defined and Y is --CH.dbd.NNHCOCH.sub.3 or ##STR46## and reacting this compound with sodium cyanoborohydride or sodium borohydride, in an aqueous buffer at a pH of 4.0 to 6.5, at a temperature of 4.degree. to 40.degree. C., inclusive, to generate a compound of formula:
- ##STR47## wherein Tu, Sp, W, m, and n are as hereinabove defined, Z is --NH--CH.sub.2 --, --CONHNHCH.sub.2 --, --NHCONHNHCH.sub.2 --, or --NHCSNHNHCH.sub.2 --, and Y is --CH.sub.2 NHNHCOCH.sub.3 or ##STR48##
A process for preparing the targeted derivatives ##STR58## of compounds of formula CH.sub.3 SSS-W wherein CH.sub.3 SSS-W is an N-acyl derivative of an antitumor antibiotic LL-E33288 .alpha..sub.2.sup.Br, .alpha..sub.2.sup.I, .beta..sub.1.sup.Br, .beta..sub.1.sup.I, .gamma..sub.1.sup.Br, .gamma..sub.1.sup.I, .delta..sub.1.sup.I, BBM-1675, FR-900405, FR-900406, PD 114759, PD 115028 CL-1577A, CL-1577B, CL-1577D, CL-1577E wherein W is ##STR59## each of R.sub.6 and R.sub.7 independenty is H or ##STR60## where R.sub.5 is CH.sub.3, C.sub.2 H.sub.5, or (CH.sub.3).sub.2 CH;
- X is an iodine or bromine atom;
- R.sub.5 is the group wherein R is hydrogen or a branched or unbranched alkyl (C.sub.1 -C.sub.10) or alkylene (C.sub.1 -C.sub.10) group, an aryl or heteroaryl group, or an aryl-alkyl (C.sub.1 -C.sub.5) or heteroaryl-alkyl (C.sub.1 -C.sub.5) group, all optionally substituted by one or more hydroxy, amino, carboxy, halo, nitro, lower (C.sub.1 -C.sub.3) alkoxy, or lower (C.sub.1 -C.sub.5) thioalkoxy groups, comprising
- reacting the compound of formula Q-Sp-SS-W, wherein Sp is a straight or branched-chain divalent or trivalent (C.sub.1 -C.sub.18 radical, divalent or trivalent (C.sub.6 -C.sub.11) aryl or heteroaryl radical, divalent or trivalent (C.sub.3 -C.sub.18) cycloalkyl or heterocycloalkyl radical, divalent or trivalent (C.sub.6 -C.sub.11) aryl or heteroaryl-alkyl (C.sub.4 -C.sub.18) radical, divalent or trivalent (C.sub.3 -C.sub.18) cycloalkyl or heterocycloalkyl-alkyl (C.sub.4 -C.sub.18) radical, divalent or trivalent (C.sub.2 -C.sub.18) unsaturated alkyl radical, wherein heteroaryl is (4-methyl-coumarin-7-yl)amino and wherein if Sp is a trivalent radical, it can be additionally substituted by amino, (C.sub.1 -C.sub.10) alkylamino, (C.sub.6 -C.sub.11) arylamino, heteroarylamino, carboxyl, lower alkoxy, hydroxy, thiol, or lower alkylthio groups;
- and Q is --CONHNH.sub.2, with nitrous acid in aqueous acetonitrile to generate a compound of formula Q-Sp-SS-W, wherein Sp and W are as hereinbefore defined and Q is --CON.sub.3 with a compound of formula Tu-(Y).sub.n, wherein Tu is a mono- or polyclonal antibody, its tumor-associated antigen-binding fragments, its chemically or genetically manipulated tumor-associated antigen-binding counterparts, or growth factors or steroids;
- Y is a side-chain amino functionality;
- and n is 1-100, to produce a compound of the formula ##STR61## wherein Tu, Z, Sp, W, m, Y, and n are as hereinabove defined.
A process for preparing the targeted derivatives ##STR62## of compounds of formula CH.sub.3 SSS-W wherein CH.sub.3 SSS-W is an N-acyl derivative of an antitumor antibiotic LL-E33288 .alpha..sub.a.sup.Br, .alpha..sub.2.sup.I, .beta..sub.1.sup.Br, .beta..sub.1.sup.I, .gamma..sub.1.sup.Br, .gamma..sub.1 BBM-1675, FR-900405, FR-900406, PD 114759, PD 115028, CL-1577A, CL-1577B, CL-1577D, CL-1577E or CL-1724 wherein W is ##STR63## each of R.sub.6 and R.sub.7 independently is H or ##STR64## where R.sub.5 is CH.sub.3, C.sub.2 H.sub.5, or (CH.sub.3).sub.2 CH;
- X is an iodine or bromine atom;
- R.sub.5' is the group RCO wherein R is hydrogen or a branched or unbranched alkyl (C.sub.1 -C.sub.10) or alkylene (C.sub.1 -C.sub.10) group, an aryl or heteroaryl group, or an aryl-alkyl (C.sub.1 -C.sub.5) or heteroaryl-alkyl (C.sub.1 -C.sub.5) group, all optionally substituted by one or more hydroxy, amino, carboxy, halo, nitro, lower (C.sub.1 -C.sub.3) alkoxy, or lower (C.sub.1 -C.sub.5) thioalkoxy groups, comprising
- reacting the compound of formula Q-Sp-SS-W, wherein Sp is a straight or branched-chain divalent or trivalent (C.sub.1 -C.sub.18) radical, divalent or trivalent (C.sub.6 -C.sub.11) aryl or heteroaryl radical, divalent or trivalent (C.sub.3 -C.sub.18) cycloalkyl or heterocycloalkyl radical, divalent or trivalent heterocycloalkyl-alkyl (C.sub.4 -C.sub.18) radical, divalent or trivalent (C.sub.2 -C.sub.18) unsaturated alkyl radical, wherein heteroaryl is (4-methyl-coumarin-7-yl)amino and wherein if Sp is a trivalent radical, it can be additionally substituted by amino, (C.sub.1 -C.sub.10) alkylamino, (C.sub.6 -C.sub.11) arylamino, heteroarylamino, carboxyl, lower alkoxy, hydroxy, thiol, or lower alkylthio groups;
- and Q is hydroxy, with an alpha-haloacetic anhydride to produce a compound wherein Q is .alpha.-haloacetyloxy, and reacting the .alpha.-haloacetyloxy-Sp-SS-W, or a compound of formula Q-SP-SS-W, wherein Sp and W are as hereinbefore defined and Q is ##STR65## with a molecule of the formula Tu-(Y).sub.n wherein Tu is a mono- or polyclonal antibody, its tumor-associated antigen-binding fragments, its chemically or genetically manipulated tumor-associated antigen-binding counterparts, or growth factors or steroids;
- Y is a side-chain thiol of a protein, or an amidoalkylthio group introduced on an amine of Tu using reagents for introducing thiol groups followed by reduction with an agent which generates the thiol group, above, or an amidoalkylthio group introduced on an amine of Tu using 2-iminothiolane, and n is 1-10, under aqueous buffered conditions at a pH between 4.5 and 7, at a temperature between 4.degree. and 40.degree. C., inclusive, to produce a compound of the formula:
- ##STR66## wherein Tu, Sp, W, and n are as hereinbefore defined, and Z is formed from covalent reaction of the Q and Y and Z is ##STR67##
Dependencies are inferred from the claim text.