Diarylsulfide backbone containing photolabile protecting groups
The present disclosure relates to photoactivable protecting groups containing a diarylsulfide chromophore, a method for the synthesis thereof and their use as photoactivable protecting groups using maskless photolithography based array synthesis.
1. A method for preparing a diarylsulfide backbone containing one or more photolabile protecting group(s) according to Formula Ib
wherein Y is S or O, and
A is selected from the group consisting of —CH 2 , —CH 2 —CH 2 —, —CH(CH 3 )—, —CH(CH 3 )—CH 2 —, and
R1 is an unsubstituted or substituted aryl- or heteroaryl-group, and R3 is H, a methyl group or an ethyl group, and
wherein R2 is
or wherein R2 is
wherein R4 is H, or R4 with the —O— of Formula III it connects to is a phosphoramidite, H-phosphonate or phosphate triester, and
wherein R5 is H, OH, a halogen or XR6, wherein X is O or S and R6 is H, an alkyl-group, aryl-group, or R6 with the —O— of Formula III it connects to is a phosphoramidite, phosphodiester, phosphotriester, H-phosphonate or an acetal or silicone moiety, and
wherein B is selected from the group consisting of adenine, cytosine, guanine, thymine, uracil, 2,6-diaminopurine-9-yl, hypoxanthin-9-yl, 5-methylcytosinyl-1-yl, 5-amino-4-imidazolecarboxylic acid-1-yl or 5-amino-4-imidazolecarboxylic acid amide-3-yl, wherein when B is adenine, cytosine or guanine the primary amino group optionally has a protecting group or when B is thymine or uracil at the O 4 position is optionally a protecting group, or
wherein R 2 is
wherein R7 is a natural amino acid, a non-natural amino acid or an amino acid derivative, including but not limited to a- or β-amino acids, and formula Ib comprises a urethane bond consisting of a N-terminal nitrogen of R7, the C═O of Formula IV that the N-terminal nitrogen of R7 connects to, and the —O— of formula Ib that the C═O of Formula IV connects to,
or wherein R2 with the —O— of formula Ib it connects to is a natural amino acid, a non-natural amino acid or an amino acid derivative, forming an ester bond to formula Ib, including but not limited to α- or β-amino acids, attached to A by an ester bond comprising the —O— of formula Ib and a C-terminal C═O of R2,
the method comprising the steps of
a) providing p-diethylbenzene as a starting material;
b) bromination of the phenylring;
c) nitration of the obtained compound in Nitric- and Sulfuric Acid in the position para- to the Bromine;
d) purification and crystallization;
e) hydroxymethylation of the compound at the benzylic position;
f) conversion of the aromatic bromine group to the arylsulfide using thiophenol;
g) purification;
h) conversion of the alcohol to chlorocarbonate; and
i) reaction of the chlorocarbonate with a nucleoside and reaction of the nucleoside with a phosphitylating agent, or
reaction of the chlorocarbonate with an amino acid derivative.
2. The method according to claim 1 , characterized in that R1 is a phenyl-group, a tert-butyl-phenyl group, a 1- or 2-naphthyl-group or a 2-pyridyl-group.
3. The method according to claim 1 , characterized in that A is —CH(CH 3 )—CH 2 —.
4. The method according to claim 1 , characterized in that R3 is H or an ethyl group.
5. A method of forming an array through photolithography, said method comprising contacting a substrate having a site activated by exposure to light with a compound of Formula Ib
wherein Y is S or O, and
A is selected from the group consisting of —CH 2 —, —CH 2 —CH 2 —, —CH(CH 3 )—, —CH(CH 3 )—CH 2 —, and
R1 is an unsubstituted or substituted aryl- or heteroaryl-group, and R3 is H, a methyl group or an ethyl group, and
wherein R2 is
or wherein R2 is
wherein R4 is H, or R4 with the —O— of Formula III it connects to is a phosphoramidite, H-phosphonate or phosphate triester, and
wherein R5 is H, OH, a halogen or XR6, wherein X is O or S and R6 is H, an alkyl-group, aryl-group, or R6 with the —O— of Formula III it connects to is a phosphoramidite, phosphodiester, phosphotriester, H-phosphonate or an acetal or silicone moiety, and
wherein B is selected from the group consisting of adenine, cytosine, guanine, thymine, uracil, 2,6-diaminopurine-9-yl, hypoxanthin-9-yl, 5-methylcytosinyl-1-yl, 5-amino-4-imidazolecarboxylic acid-1-yl or 5-amino-4-imidazolecarboxylic acid amide-3-yl, wherein when B is adenine, cytosine or guanine the primary amino group optionally has a protecting group or when B is thymine or uracil at the 0 4 position is optionally a protecting group, or
wherein R2 is
wherein R7 is a natural amino acid, a non-natural amino acid or an amino acid derivative, including but not limited to a- or β-amino acids, and formula Ib comprises a urethane bond consisting of a N-terminal nitrogen of R7, the C═O of Formula IV that the N-terminal nitrogen of R7 connects to, and the —O— of formula Ib that the C═O of Formula IV connects to,
or wherein R2 with the —O— of formula Ib it connects to is a natural amino acid, a non-natural amino acid or an amino acid derivative, forming an ester bond to formula Ib, including but not limited to α- or β-amino acids, attached to A by an ester bond comprising the —O— of formula Ib and a C-terminal C═O of R2.
6. The method of claim 5 wherein the contacting of the substrate with a compound of Formula Ib produces a DNA microarray or a peptide microarray.
7. The method of claim 6 wherein the method produces a DNA microarray.
8. The method of claim 6 wherein the method produces a peptide microarray.
9. The method of claim 5 wherein the substrate and/or the compound is activated by exposure to light between the wavelengths of 374 to 405 nm.