144095-63-6

This paper deals with the synthesis of a stable biotin-phosmidosine conjugate molecule 3 that is required for isolation of biomolecules that bind to phosmidosine (1). It was found that introduction of a biotin residue into the 6-N position of phosmidosine could be carried out by reaction of an N 7-Boc-7,8-dihydro-8-oxoadenosine derivative 13 with phenyl chloroformate followed by displacement with a diamine derivative 6 along with the simultaneous removal of the Boc group and one of the two phenoxycarbonyl groups and the successive condensation with an N-tritylated biotin derivative 5. The condensation of an N-prolylphosphorodiamidite derivative 4 with an appropriately protected 7,8-dihydro-8-oxoadenosine derivative 17 having the biotin residue gave the coupling product 18, which was deprotected to give the biotin-phosmidosine (O-ethyl ester) conjugate 3.

The synthesis of 1-phenylpyrazolin-5-one carboxylate esters of biotin and fluorescein and their specific reactivity with amines under mild conditions to label various synthetic biopolymers such as oligonucleotides, peptides and PNA's is described.

The invention discloses a long-chain biotin marker and a preparation method and application thereof. The long-chain biotin marker comprises a biotin molecule, a spacer arm and a reaction group. The biotin molecule comprises a ring structure and a valeric acid side chain, a structure formula (I) of the spacer arm is as shown in the specification, a structural formula (II) of the reaction group is as shown in the specification, the valeric acid side chain in the biotin molecule is connected with NH at a first tail end of the spacer arm, oxygen at a second tail end of the spacer arm is combined with phosphorus in the reaction group to form an O-P bond, and accordingly connection between the spacer arm and the reaction group is realized. The long-chain biotin marker is used for gene capturingand sequencing.

(Chemical Equation Presented) CCA-pcb (cytidylyl-(3′5′)- cytidylyl-(3′5′)-3′(2′)-O-(N-(6-D-(+) -biotinoylaminohexanoyl)-L-phenylalanyl)adenosine), a ribosomal P-site substrate, was synthesized by phosphoramidite chemistry in 26 steps with an overall yield