Bio-orthogonal drug activation
The invention relates to a Prodrug activation method, for therapeutics, wherein use is made of abiotic reactive chemical groups that exhibit bio-orthogonal reactivity towards each other. The invention also relates to a Prodrug kit comprising at least one Prodrug and at least one Activator, wherein the Prodrug comprises a Drug and a first Bio-orthogonal Reactive Group (the Trigger), and wherein the Activator comprises a second Bio-orthogonal Reactive Group. The invention also relates to targeted therapeutics used in the above-mentioned method and kit. The invention particularly pertains to antibody-drug conjugates and to bi- and trispecific antibody derivatives.
1. A drug-dienophile conjugate of formula (1a):
which comprises a drug D D and a dienophile moiety;
wherein T and G each independently denotes H, or a substituent selected from the group consisting of alkyl, F, Cl, Br or I;
A and P each independently are CR a 2 or CR a X D , provided that at least one is CR a X D ; X D is (O—C(O)) p -(L D ) n -(D D ), S—C(O)-(L D ) n -(D D ), O—C(S)-(L D ) n -(D D ), S—C(S)-(L D ) n -(D D ) wherein p=0 or 1; L D is a self-immolative linker; n=0 or 1; each D D independently is a drug;
Y, Z, Q, X together form a four-membered aliphatic or heteroaliphatic moiety, optionally fused to an aromatic moiety or moieties;
each R a independently is selected from the group consisting of H, alkyl, aryl, OR′, SR′, S(═O)R′″, S(═O) 2 R′″, S(═O) 2 NR′R″, F, Cl, SO 3 H, SO 4 H, NO 2 , CN, CF 3 , CF 2 —R′, NR′R″, C(═O)R′, C(═O)OH, C(═O)NR′R″, C(═S)NR′R″, NR′C(═O)—R′″, NR′C(═S)—R′″, NR′C(═O)O—R′″, NR′C(═S)O—R′″, NR′C(═O)S—R′″, OC(═O)NR′—R′″, SC(═O)NR′—R′″, OC(═S)NR′—R′″, SC(═S)NR′—R′″, NR′C(═S)S—R′″, NR′C(═O)NR″—R″, and NR′C(═S)NR″—R″, with each R′ and each R″ independently being H, aryl or alkyl and R′″ independently being aryl or alkyl;
wherein L D is linked to R T via S, N, NH, or O, wherein these atoms are part of the linker;
wherein D D is one or more drugs, linked to L D , O—C(O), S—C(O), O—C(S), S—C(S) or CR a via S, N, NH, or O, wherein these atoms are part of the drug; and wherein when X D is (O—C(O)) p -(L D ) n -(D D ) and p=0 and L D or D D is bound to T R via N or NH, these N and NH moieties are bound to an aromatic carbon of L D or D D ; and
wherein optionally one or more targeting agents T T or masking moieties M M are attached to the Drug D D , the dienophile moiety, or Linker L D , optionally via a spacer or spacers S P .
2. A drug-dienophile conjugate according to claim 1 , wherein L D is linked to the dienophile moiety via S, N, NH, or O, wherein these atoms are part of the linker.
3. A drug-dienophile conjugate according to claim 1 , wherein L D consists of multiple units arranged linearly and/or branched.
4. A drug-dienophile conjugate according to claim 1 , wherein D D is linked via S, N, NH, or O, wherein these atoms are part of the drug.
5. A drug-dienophile conjugate according to claim 1 , wherein X D is
(O—C(O)) p -(L D ) n -(D D ), wherein p=0 or 1, and n=0 or 1.
6. A drug-dienophile conjugate according to claim 1 , wherein the drug-dienophile conjugate satisfies formula (1b):
wherein each R a independently denotes H, or, in at most four instances, a substituent selected from the group consisting of alkyl, aryl, OR′, SR′, S(═O)R′″, S(═O) 2 R′″, S(═O) 2 NR′R″, Si—R′″, Si—O—R′″, OC(═O)R′″, SC(═O)R′″, OC(═S)R′″, SC(═S)R′″, F, Cl, Br, I, N 3 , SO 2 H, SO 3 H, SO 4 H, PO 3 H, PO 4 H, NO, NO 2 , CN, OCN, SCN, NCO, NCS, CF 3 , CF 2 —R′, NR′R″, C(═O)R′, C(═S)R′, C(═O)O—R′, C(═S)O—R′, C(═O)S—R′, C(═S)S—R′, C(═O)NR′R″, C(═S)NR′R″, NR′C(═O)—R′″, NR′C(═S)—R′″, NR′C(═O)O—R′″, NR′C(═S)O—R′″, NR′C(═O)S—R′″, NR′C(═S)S—R′″, OC(═O)NR′—R′″, SC(═O)NR′—R′″, OC(═S)NR′—R′″, SC(═S)NR′—R′″, NR′C(═O)NR″—R″, NR′C(═S)NR″—R″, CR′NR″, with each R′ and each R″ independently being H, aryl or alkyl and R′″ independently being aryl or alkyl;
each R d as above indicated is independently selected from the group consisting of H, alkyl, aryl, OR′, SR′, S(═O)R′″, S(═O) 2 R′″, Si—R′″, Si—O—R′″, OC(═O)R′″, SC(═O)R′″, OC(═S)R′″, SC(═S)R′″, F, Cl, Br, I, N 3 , SO 2 H, SO 3 H, PO 3 H, NO, NO 2 , CN, CF 3 , CF 2 —R′, C(═O)R′, C(═S)R′, C(═O)O—R′, C(═S)O—R′, C(═O)S—R′, C(═S)S—R′, C(═O)NR′R″, C(═S)NR′R″, NR′C(═O)—R′″, NR′C(═S)—R′″, NR′C(═O)O—R′″, NR′C(═S)O—R′″, NR′C(═O)S—R′″, NR′C(═S)S—R′″, NR′C(═O)NR″—R″, NR′C(═S)NR″—R″, CR′NR″, with each R′ and each R″ independently being H, aryl or alkyl and R′″ independently being aryl or alkyl; wherein two R a,d moieties together may form a ring;
wherein, additionally, at most two exocyclic bonds fixed in the same plane are optionally present; with optionally one or more R a,d comprised in a linker moiety, optionally via a spacer S P , to a targeting agent T T or a masking moiety M M , and wherein T and G each independently denote H, or a substituent selected from the group consisting of alkyl, F, Cl, Br, and I, and X D is as defined for formula (1a) in claim 1 .
7. A drug-dienophile conjugate according to claim 1 selected from the following structures:
wherein T T denotes a targeting agent, S P denotes a spacer or spacers, and M M denotes a masking moiety.
8. A drug-dienophile conjugate according to claim 1 , wherein one or more targeting agents T T is attached to the drug D D , the linker L D , or the trigger moiety T R , optionally via a spacer or spacers S P .
9. A drug-dienophile conjugate according to claim 1 , wherein T T is an antibody, antibody fragment, protein, peptide, peptide mimetic, polymer, carbohydrate, or oligonucleotide targeting agent.
10. A drug-dienophile conjugate according to claim 1 , wherein one or more masking moieties M M is attached to the drug D D , the linker L D , or the dienophile moiety, optionally via a spacer or spacers S P .
11. A drug-dienophile conjugate according to claim 1 , wherein the masking moiety is a protein, peptide, polymer, polyethyleneglycol, or carbohydrate.
12. A drug-dienophile conjugate according to claim 1 , wherein the drug D D is selected from the group consisting of proteins, oligopeptides, oligonucleotides, oligosaccharides, peptides, peptoids, and organic drug compounds.
13. A drug-dienophile conjugate according to claim 1 , wherein the drug D D is a T-cell engaging antibody construct.
14. A drug-dienophile conjugate according to claim 1 , wherein the drug D D is selected from the group consisting of antiproliferative agents, antitumor agents, antibiotics, anti-inflammatory agents, anti-viral agents, antihypertensive agents, chemo sensitizing agents, radiation sensitizers, cytotoxic drugs, and differentiation inducers.
15. The drug-dienophile conjugate according to claim 1 , wherein the drug D D is selected from the group consisting of anti-tubulin agents, topoisomerases inhibitors, dihydrofolate reductase inhibitors, thymidylate synthase inhibitors, DNA minor groove binders, DNA damaging agents, DNA intercalators, DNA cleavers, and DNA alkylators.
16. The drug-dienophile conjugate according to claim 1 , wherein the drug D D is selected from the group consisting of cytokines, toxins, immunotoxins, hormones, steroids, platinum-based drugs, cytotoxic nucleosides, taxanes, lexitropsins, pteridines, diynenes, podophyllotoxins, dolastatins, maytansinoids, anthracyclines, vinca drugs, mitomycins, bleomycins, taxols, ricin A toxins, diphtheria toxins, and cholera toxins.
17. A drug-dienophile conjugate according to claim 1 , wherein the prodrug is an antibody-toxin or an antibody-drug conjugate.
18. A drug-dienophile conjugate according to claim 1 , wherein:
A is CR a X D with R a being H and X D being in the axial position, and P is CR a 2 with R a being H, and T and G are H; or wherein
A is CR a X D with R a being H and X D being in the axial position, and P is CR a 2 with R a being H, T and G are H, Y, Q, X, Z are CR a 2 ; or wherein
A is CR a X D with R a being H and X D being in the axial position, and P is CR a 2 with R a being H, T and G are H, and Y, Q, X, Z are CR a 2 wherein in at most two instances R a is not a hydrogen.
19. A drug-dienophile conjugate according to claim 1 , wherein T, G and R a are H, and X D is in the axial position and is (O—C(O)) p -(L D ) n -(D D ), wherein p is 0 or 1, and n is 0 or 1.
20. A method of administering a drug to a patient suffering from a disease that can be modulated by the drug, the method comprising:
(a) administering to said patient a drug-dienophile conjugate having formula (1a) of claim 1 ; and
(b) releasing the drug from the drug-dienophile conjugate by administering an activator, wherein the activator comprises a diene selected from the dienes according to Formula 4
wherein R 1 and R 2 each independently are selected from the group consisting of H, alkyl, aryl, NR′C(═O)R″, NR′C(═S)R″, NR′C(═O)OR″, NR′C(═S)OR″, NR′C(═O)SR″, NR′C(═S)SR″, OC(═O)NR′R″, SC(═O)NR′R″, OC(═S)NR′R″, SC(═S)NR′R″, NR′C(═O)NR″R″, NR′C(═S)NR″R″ with each R′ and each R″ independently being H, aryl or alkyl, and R′″ independently being aryl or alkyl;
wherein A is N; B is N; X is N; and Y is N;
wherein neither or at least one of the substituents R 1 and R 2 on the diene is comprised in a binding to a polymer, protein, peptide, carbohydrate, dendrimer, heparin derivative, hyaluronic acid derivative, albumin, albumin-binding moiety, dye moiety, fluorescent moiety, imaging probe, chelate, liposome, polymer particle or polymersome.
21. A method according to claim 20 , wherein one or more targeting agents T T is attached to the drug D D , the linker L D , or the trigger moiety T R , optionally via a spacer or spacers S P .
22. A method according to claim 21 , wherein T T is an antibody, antibody fragment, protein, peptide, peptide mimetic, polymer, carbohydrate, or oligonucleotide targeting agent.
23. A method according to claim 20 , wherein one or more masking moieties M M is attached to the drug D D , the linker L D , or the dienophile moiety, optionally via a spacer or spacers S P .
24. A method according to claim 20 , wherein the masking moiety is a protein, peptide, polymer, polyethyleneglycol, or carbohydrate.
25. A method according to claim 20 , wherein the drug D D is selected from the group consisting of proteins, oligopeptides, oligonucleotides, oligosaccharides, peptides, peptoids, and organic drug compounds.
26. A method according to claim 20 , wherein the drug D D is a T-cell engaging antibody construct.
27. A method according to claim 20 , wherein the drug D D is selected from the group consisting of antiproliferative agents, antitumor agents, antibiotics, anti-inflammatory agents, anti-viral agents, antihypertensive agents, chemo sensitizing agents, radiation sensitizers, cytotoxic drugs, and differentiation inducers.
28. A method according to claim 20 , wherein the drug D D is selected from the group consisting of anti-tubulin agents, topoisomerases inhibitors, dihydrofolate reductase inhibitors, thymidylate synthase inhibitors, DNA minor groove binders, DNA damaging agents, DNA intercalators, DNA cleavers, and DNA alkylators.
29. A method according to claim 20 , wherein the drug D D is selected from the group consisting of cytokines, toxins, immunotoxins, hormones, steroids, platinum-based drugs, cytotoxic nucleosides, taxanes, lexitropsins, pteridines, diynenes, podophyllotoxins, dolastatins, maytansinoids, anthracyclines, vinca drugs, mitomycins, bleomycins, taxols, ricin A toxins, diphtheria toxins, and cholera toxins.
30. A method according to claim 20 , wherein the drug-dienophile conjugate is an antibody-toxin or an antibody-drug conjugate.
31. A method according to claim 20 , wherein:
A in formula (1a) is CR a X D with R a being H and X D being in the axial position, and P is CR a 2 with R a being H, and T and G are H; or wherein
A in formula (1a) is CR a X D with R a being H and X D being in the axial position, and P is CR a 2 with R a being H, T and G are H, Y, Q, X, Z are CR a 2 ; or wherein
A in formula (1a) is CR a X D with R a being H and X D being in the axial position, and P is CR a 2 with R a being H, T and G are H, and Y, Q, X, Z are CR a 2 wherein in at most two instances R a is not a hydrogen.
32. A method according to claim 20 , wherein T, G and R a are H, and X D is in the axial position and is (O—C(O)) p -(L D ) n -(D D ), wherein p is 0 or 1, and n is 0 or 1.
33. A drug-dienophile conjugate according to claim 1 , wherein when a targeting agent T T is present, it is an organic compound.
34. A method of administering a drug to a patient suffering from a disease that can be modulated by the drug, the method comprising administering to the patient a drug-dienophile conjugate and an activator;
wherein the drug-dienophile conjugate has formula (1a):
which comprises the drug D D and a dienophile moiety;
wherein T and G each independently denotes H or a substituent selected from the group consisting of alkyl, F, Cl, Br or I;
A and P each independently are CR a 2 or CR a X D , provided that at least one is CR a X D ;
X D
is (O—C(O)) p -(L D ) n -(D D ), S—C(O)-(L D ) n -(D D ), O—C(S)-(L D ) n -(D D ), or S—C(S)-(L D ) n -(D D )
wherein p=0 or 1; L D is a self-immolative linker; n=0 or 1; each D D independently is a drug;
Y, Z, Q, X together form a four-membered aliphatic or heteroaliphatic moiety, optionally fused to an aromatic moiety or moieties;
each R a independently is selected from the group consisting of H, alkyl, aryl, OR′, SR′, S(═O)R′″, S(═O) 2 R′″, S(═O) 2 NR′R″, F, Cl, SO 3 H, SO 4 H, NO 2 , CN, CF 3 , CF 2 —R′, NR′R″, C(═O)R′, C(═O)OH, C(═O)NR′R″, C(═S)NR′R″, NR′C(═O)—R″, NR′C(═S)—R′″, NR′C(═O)O—R′″, NR′C(═S)O—R′″, NR′C(═O)S—R′″, OC(═O)NR′—R′″, SC(═O)NR′—R′″, OC(═S)NR′—R′″, SC(═S)NR′—R′″, NR′C(═S)S—R′″, NR′C(═O)NR″—R″, and NR′C(═S)NR″—R″, with each R′ and each R″ independently being H, aryl or alkyl and R′″ independently being aryl or alkyl;
wherein L D is linked to T R via S, N, NH, or O, wherein these atoms are part of the linker; wherein D D is one or more drugs, linked to L D , O—C(O), S—C(O), O—C(S), S—C(S) or CR a via S, N, NH, or O, wherein these atoms are part of the drug; and wherein when X D is (O—C(O)) p -(L D ) n -(D D ) and p=0 and L D or D D is bound to T R via N or NH, these N and NH moieties are bound to an aromatic carbon of L D or D D ; and
wherein optionally one or more targeting agents T T or masking moieties M M are attached to the Drug D D , the dienophile moiety, or Linker L D , optionally via a spacer or spacers S P ;
and wherein the activator has Formula 4
wherein R 1 and R 2 each independently are selected from the group consisting of H, alkyl, aryl, NR′C(═O)R″, NR′C(═S)R″, NR′C(═O)OR″, NR′C(═S)OR″, NR′C(═O)SR″, NR′C(═S)SR″, OC(═O)NR′R″, SC(═O)NR′R″, OC(═S)NR′R″, SC(═S)NR′R″, NR′C(═O)NR″R″, NR′C(═S)NR″R″ with each R′ and each R″ independently being H, aryl or alkyl, and R′″ independently being aryl or alkyl;
wherein A is N; B is N; X is N; and Y is N;
wherein neither or at least one of the substituents R 1 and R 2 on the diene is comprised in a binding to a polymer, protein, peptide, carbohydrate, dendrimer, heparin derivative, hyaluronic acid derivative, albumin, albumin-binding moiety, dye moiety, fluorescent moiety, imaging probe, chelate, liposome, polymer particle or polymersome.