Patent ID: 8729243

Claim:
Method for preparing a compound having the formula (I): wherein X is biotin or Y is biotin or Z is biotin or V is biotin or B is NH 2 or H, AA is a trifunctional molecule derivative when B is NH 2 or a bifunctional molecule derivative when B is H, AA 1 to AA 5 are each independently a trifunctional molecule derivative, G 0 to G 5 are each independently an arm comprising at least one (—CH 2 —CH 2 —O) unit, n 0 to n 5 are each independently a whole number between 1 and 8 and T is an antiligand or a reactive group for fixation to an antiligand, said antiligand being capable of reacting with a ligand, the method comprising: (i) grafting n 0 compounds having the formula W″ 0 -G 0 -OH, where W″ 0 is an amine protecting group, n 0 and G 0 being such as defined above, to a compound having the formula (II): where R is a prefunctionalised resin, W and W 0 are different from one another and represent an amine protecting group, W being different from W″ 0 , and AA is such as defined above, to obtain a compound having the formula (III): (ii) coupling a compound having a formula (IV): where W 1 and W′ 1 are amine protecting groups that are identical to or different from one another and from the W 0 and W″ 0 groups, but different from W, and AA 1 is such as defined above, with the compound having the formula (III) to obtain a compound having the formula (V): (iii) grafting 2n 1 compounds having the formula W″ 1 -G 1 -OH, where W″ 1 is an amine protecting group, identical to or different from one another and from the other protecting groups used in this method, but different from W, G 1 and n 1 being such as defined above, onto the compound obtained in step (ii) to obtain a compound having the formula (VI): (iv) when X in the formula (I) is biotin, passing directly to step (v); if not, when X in the formula (I) is not biotin, repeating the steps (ii) and (iii) with 2 p-1 compounds having the formula (VII): and 2 p n p times the compound having the formula W″ p -G p -OH, where p is a whole number between 2 and 5, and W p , W′ p , W″ p are amine protecting groups that are identical to or different from one another and from the other protecting groups used in this method, but different from W, and AA p is a trifunctional molecule, according to the following sequence: 1 time when Y is biotin, p then being equal to 2 2 times when Y is not biotin and Z is biotin, p then being equal to 2 then to 3, 3 times when Y and Z are not biotin and V is biotin, p then being equal to 2, 3 then 4, and 4 times when Y, Z and V are not biotin, p then being equal to 2, 3, 4 then 5, (v) deprotecting the compound thus obtained at the W″ 1 or W″ p group, p being between 2 and 5, and coupling with biotin, (vi) deprotecting, at the W group, the compound thus polybiotinylated and coupling with an antiligand or a reactive group for fixation to an antiligand (T) and (vii) cutting the compound thus obtained from the resin (R) to obtain a compound having the formula (I).