Chemically cleavable group

1. A method for provoking the release, in vitro, of a construct attached to a chemically cleavable group, wherein the chemically cleavable group is a dienophile, by allowing the dienophile to react with a diene, wherein the dienophile is an eight-membered non-aromatic cyclic alkenylene group, wherein the dienophile satisfies Formula (1a):

wherein A and P each independently are CH 2 or CHX D , provided that at least one is CHX D ; X D is (O—C(O)) p -(L D ) n -(C A ), or O—C(S)-(L D ) n -(C A ), wherein n=0 or 1, wherein p=0 or 1, wherein when p=0, L D or C A is bound to the dienophile via O, S, aromatic N or aromatic NH, Y, Z, X, Q each independently are selected from the group consisting of CR a 2 , C═O, O, and NR b , with at most one of Y, Z, X, and Q being C═O, wherein two R moieties together may form a ring, and with the proviso that no adjacent pairs of atoms are present selected from the group consisting of O—O, and O—NR b ; and wherein one R a,b is optionally bound, directly or via a spacer or spacers S P , to a construct C B ; and

wherein the self-immolative linker L D , in addition to being bound to C A , is also optionally bound, directly or via a spacer or spacers S P , to a construct C B ; and wherein T and G denote H; wherein each R a can independently be H, alkyl, aryl, OR′, SR′, S(═O) 2 R′″, S(═O) 2 NR′R″, SO 3 H, PO S H, NO 2 , CN, CF 3 , CF 2 —R′, NR′R″, C(═O)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″, or NR′C(═S)NR″—R″, with each R′ and each R″ independently being H, aryl or alkyl and R′″ independently being aryl or alkyl; each R b is independently selected from the group consisting of H, alkyl, aryl, O-aryl, O-alkyl, OH, and C(═O)NR′R″ with R′ and R″ each independently being H, aryl or alkyl, R′CO-alkyl with R′ being H, alkyl, and aryl; wherein C A and C B are each individually a small molecule, an organic molecule, a metal coordination compound, an inorganic molecule, an organometallic molecule, a biomolecule, a drug, a polymer, a resin, a particle, a gel, a surface, a cell, a biological tissue, a pathogen, or a combination thereof; or wherein either or both of C A and C B is present in a binding to a biomolecule; or wherein either or both of C A and C B is comprised in a binding to a polymer, resin, particle, or solid support; or wherein the construct comprises a biomolecule binding moiety, selected from the group consisting of non-covalent binding moieties, and covalent binding moieties;

wherein the diene satisfies formula (4):

wherein R 1 and R 2 each independently are selected from the group consisting of H, alkyl, aryl, S(═O) 2 R′″, S(═O) 2 OR′, C(═O)NR′R″, C(═S)NR′R″, NR′C(═O)OR″, NR′C(═S)OR″, NR′C(═O)SR″, NR′C(═S)SR″, NR′C(═O)NR″R″, and NR′C(═S)NR″R″ with each R′ and each R″ independently being H, aryl or alkyl; with A, B, X, and Y being N; and wherein the diene optionally has a link to a moiety selected from the group consisting of a peptide, a protein, a carbohydrate, a polymer, a resin, a particle, a gel, and a surface.

2. The method according to claim 1 , wherein the dienophile satisfies Formula (1a):

wherein T and G are H, and further wherein A and P each independently are CH 2 or CHX D , provided that at least one is CHX D , X D is (O—C(O)) p -(L D ) n -(C A ), or O—C(S)-(L D ) n -(C A ), wherein p=0 or 1 and n=0 or 1, wherein when p=0, L D or C A is bound to the dienophile via O, S, aromatic N or aromatic NH; wherein one of the bonds PQ, QX, XZ, ZY, YA is part of a fused ring, such that two exocyclic bonds are fixed in the same plane, and provided that PQ and YA are not part of an aromatic 5- or 6-membered ring, or of a conjugated 7-membered ring; when not part of a fused ring P and A are independently CH 2 or CHX D , provided that at least one is CHX D ; when part of a fused ring P and A are independently CH or CX D , provided that at least one is CX D ; the remaining moieties Y, Z, X, and Q of the group (—Y—Z—X-Q-) being independently from each other CR a 2 , S, SO, SO 2 , O, NR b , such that no adjacent pairs of atoms are present selected from the group consisting of O—O, O—NR b , S—NR b , O—S, O—S(O), O—S(O) 2 .

3. The method according to claim 1 , wherein the chemically cleavable group is a trans cyclooctene moiety that satisfies formula (1b):

wherein, in addition to the optional presence of at most two exocyclic bonds fixed in the same plane, each R a independently denotes H, or, in at most three instances, a substituent selected from the group consisting of alkyl, aryl, OR′, SR′, S(═O) 2 R′″, S(═O) 2 NR′R″, SO 3 H, PO 3 H, NO 2 , CN, CF 3 , CF 2 —R′, NR′R″, C(═O)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″, 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 each R e as above indicated is independently selected from the group consisting of H, alkyl, aryl, OR′, SR′, S(═O) 2 R′″, F, SO 3 H, PO 3 H, NO 2 , CN, CF 3 , CF 2 —R′, C(═O)R′, C(═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″, 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 two R a,e moieties together may form a ring; and wherein T and G denote H, and X D is (O—C(O)) p -(L D ) n -(C A ), or O—C(S)-(L D ) n -(C A ), wherein p=0 or 1.

4. The method according to claim 1 , wherein the trans-cyclooctene moiety satisfies any one of the following formulae:

and wherein S P is a spacer.

5. The method according to claim 1 , wherein the trans-cyclooctene moiety satisfies any one of the following formulae

wherein =rest of attached C A and L D -C A , wherein L D may optionally comprise C B or S P —C B ,

wherein -----=rest of attached C B or S P —C B .

6. The method according to claim 1 , wherein the trans-cyclooctene moiety satisfies any one of the following formulae

7. The method according to claim 1 , wherein the diene satisfies formula (8a) or (8b):

wherein each R 1 and each R 2 independently are selected from the group consisting of H, alkyl, aryl, CF 3 , CF 2 —R′, NO 2 , OR′, SR′, C(═O)R′, C(═S)R′, OC(═O)R′″, SC(═O)R′″, OC(═S)R′″, SC(═S)R′″, S(═O)R′, S(═O) 2 R′″, S(═O) 2 NR′R″, C(═O)O—R′, C(═O)S—R′, C(═S)O—R′, C(═S)S—R′, C(═O)NR′R″, C(═S)NR′R″, NR′R″, 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″, 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.

8. The method according to claim 1 , wherein the diene is satisfies a formula selected from the group consisting of (8c), (8d), (8e), (8f), and (8g):

wherein each R 1 and each R 2 independently are selected from the group consisting of H, alkyl, aryl, CF 3 , CF 2 —R′, NO 2 , OR′, SR′, C(═O)R′, C(═S)R′, OC(═O)R′″, SC(═O)R′″, OC(═S)R′″, SC(═S)R″, S(═O)R′, S(═O) 2 R′″, S(═O) 2 NR′R″, C(═O)O—R′, C(═O)S—R′, C(═S)O—R′, C(═S)S—R′, C(═O)NR′R″, C(═S)NR′R″, NR′R″, 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″, 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.

9. The method according to claim 1 , wherein the diene satisfies any one of the formulae:

10. The method according to claim 1 , wherein the diene satisfies any one of the formulae:

11. The method according to claim 1 , wherein the diene satisfies any one of the formulae:

12. The method according to claim 1 , wherein the diene satisfies the formula:

13. The method according to claim 1 , wherein the diene satisfies the formula:

14. The method according to claim 1 , wherein the diene satisfies the formula:

15. The method according to claim 1 , wherein the diene satisfies the formula:

16. The method according to claim 1 , wherein either or both of C A and C B is a biomolecule.

17. The method according to claim 1 , wherein either or both of C A and C B is present in a binding to a biomolecule.

18. The method according to claim 1 , wherein C A or C B is a polymer, resin, particle, solid support.

19. The method according to claim 1 , wherein C A or C B is comprised in a binding to a polymer, resin, particle, or solid support.

20. The method according to claim 1 , wherein the construct comprises, a biomolecule binding moiety, selected from the group consisting of non-covalent binding moieties, and covalent binding moieties.

21. The method according to claim 1 , wherein the dienophile serves as a bioorthogonal linker or mask for reversible biomolecule modification in vitro.

22. The method according to claim 1 wherein Y, Z, X, Q each independently are selected from the group consisting of CR a 2 and O with at most two of Y, Z, X, and Q being O, and wherein T and G are H.

23. The method according to claim 3 wherein the dienophile has a structure according to formula (1b) wherein R e and T and G are H and wherein in at most four instances R a is not a hydrogen.

24. The method according to claim 1 wherein the diene has a structure according to formula (4) wherein R 1 and R 2 each independently are selected from the group consisting of H, alkyl, aryl, wherein A is N; B is N; X is N; and Y is N.

25. The method according to claim 1 , wherein X D is (O—C(O)) p -(L D ) n -(C A ), wherein p=0 or 1, and n=0 or 1.

26. The method according to claim 20 , wherein the non-covalent binding moiety is a biomolecule, biotin or an antibody.

27. The method according to claim 20 , wherein the covalent binding moiety is an N-hydroxysuccinimide ester, a maleimide, an azide, an alkyne, or a photoreactive group.

28. The method according to claim 2 , wherein one R a,b,c or the self-immolative linker L D is bound to C B .

29. The method according to claim 28 , wherein the one R a,b,c or the self-immolative linker L D is bound, via a spacer or spacers S P , to C B .

30. The method according to claim 3 wherein one R a,e or the self-immolative linker L D is bound to C B .

31. The method according to claim 30 wherein one R a,e or the self-immolative linker L D , is bound, via a spacer or spacers S P , to C B .

32. The method according to claim 1 wherein one R a,b or the self-immolative linker L D is bound to C B .

33. The method according to claim 32 wherein one R a,b or the self-immolative linker L D is bound, via a spacer or spacers S P , to C B .

34. The method according to claim 4 wherein S P is a polyethylene glycol (PEG) chain varying from 2 to 200 repeating units or a biopolymer fragment.

35. The method according to claim 34 wherein S P is a biopolymer fragment wherein the biopolymer fragment is an oligopeptide, a polypeptide, a polylactide or a carbohydrate.

36. The method according to claim 1 wherein the diene has a link to a moiety selected from the group consisting of a peptide, a protein, a carbohydrate, a polymer, a resin, a particle, a gel, and a surface.

37. The method according to claim 36 , wherein the polymer is PEG.