Protected biotin derivatives

Claim 2

A process for the preparation of a compound of formula Ia as set out hereinafter [in which m is 4 or 5, X represents a direct link, -O-P(O)(OR⁶ a )-O-, -S-, -O-, -CONH-, -NHCONH- or N-R⁸ (wherein R⁸ represents a straight chain or branched C₁₋₁₀ alkyl group), p is an integer from 0 to 16 with the proviso that where X is other than a direct link p is an integer of at least 2, R⁶ a represents a hydrogen atom or a phosphate protecting group and Z′ represents a nucleotide sequence in unprotected form or in a form in which base and/or phosphate moieties carry protecting group(s), and the nucleotide sequence is optionally bound to a support] or an isoequivalent thereof, which process comprises deprotecting a compound of formula II as set out hereinafter [in which m, p, X, R⁶ a and Z′ are as hereinbefore defined and J and K which may be the same or different, each represents a hydrogen atom or a protecting group compatible with nucleotide phosphorylation which group increases the lipophilicity of the compound, at least one of J and K being other than hydrogen] or an isoequivalent thereof; whereby to remove any J or K group being other than hydrogen to obtain a compound of formula Ia or isoequivalent thereof.

Claim 3

A process as claimed in claim 1 wherein J and/or K represent(s) tetrahydropyranyl, furyl, dimethoxytrityl, xanthenyl, tetrahydrothiopyranyl, benzoyl or an acyl group.

Claim 4

A process as claimed in claim 1 or claim 2 wherein R⁶ a represents hydrogen, methyl, 2-cyanoethyl, 2-chlorophenyl, 2,2,2-­trihalo-1,1-dimethyl-ethyl, 5-chloroquinolin-8-yl, 2-methylthioethyl, or a 2-phenylthioethyl group optionally substituted on the phenyl ring by halogen or NO₂.

Claim 5

A process as claimed in any one of claims 1 to 3 wherein J and/or K represent(s) tetrahydropyran-2-yl, 6-methoxy-tetrahydropyran-­2-yl, or dimethoxytrityl.

Claim 6

A process as claimed in any one of the previous claims wherein only one of J and K represents a protecting group, the other of J and K representing hydrogen.

Claim 7

A process for the preparation of a compound of the formula II set out hereinafter as defined in any one of claims 1-5 or an isoequivalent thereof, which process comprises oxidising a corresponding compound of the formula III as set out hereinafter [wherein m, p, J, K, Z′, X and R⁶ a are as hereinbefore defined] or an equivalent thereof.

Claim 8

A compound of formula II set out hereinafter [in which m, p, X, R⁶ a , Z′, J and K have the meanings stated in any one of the preceding claims] or an isoequivalent thereof.

Claim 9

A process for the preparation of a compound of formula III set out hereinafter (in which J, K, m, p, X, R⁶ a and Z′ have the meanings stated in any one of claims 1 to 5 above] or an isoequivalent thereof, which process comprises coupling a compound of formula IV set out hereinafter (in which J, K, m, p and X are as defined above and R⁴ and R⁵, which may be the same or different, each represent a 1-10C straight chain or branched alkyl group or R⁴ and R⁵ together with the nitrogen atom there between represent a 5-7 membered heterocyclic ring containing 1, 2, 3 or 4 hetero atoms, any hetero atom present in addition to nitrogen being selected from oxygen, nitrogen and sulphur, and R⁶ corresponds to R⁶ as hereinbefore defined, but is other than hydrogen] or an isoequivalent thereof, with a nucleotide sequence of the formula Z˝ which corresponds to a nucleotide sequence of the formula Z′ as defined above but wherein the point of attachment to the compound of formula IV is unprotected such as to allow coupling at that point, whereby to obtain a compound of formula III as hereinbefore defined or an isoequivalent thereof.

Claim 10

A compound of formula III set out hereinafter [in which J, K, m, p, X, R⁶ a and Z¹ have the meanings stated in any one of claims 1 to 8 above] or an isoequivalent thereof.

Claim 11

A process for the preparation of a compound of formula IV set out hereinafter [in which J, K, m, p, X, R⁶, R⁴ and R⁵ have the meanings stated in any one of claims 1 to 8 above] or an isoequivalent thereof, which process comprises reacting a compound of the formula VI set out hereinafter [in which J, K, m, p, and X are as defined above] or an isoequivalent thereof with a compound of formula V set out hereinafter [wherein R⁴, R⁵ and R⁶ are as defined above, and L is a displaceable group] whereby to form a compound of formula IV or an isoequivalent thereof.

Claim 12

A compound of formula IV set out hereinafter [in which J, K, m, p, R⁴, R⁵, R⁶ and X have the meanings stated in any one of claims 1 to 8 above] or an isoequivalent thereof.

Claim 13

A process for the preparation of a compound of formula VI set out hereinafter [in which J, K, m, p and X have the meanings stated in anyone of claims 1 to 10 above] or an isoequivalent thereof which process comprises: (a) where X represents a direct link, reducing a corresponding compound of the general formula VII set out hereinafter [wherein J, K, m and p are as defined above and R represents a group reduceable to the corresponding alcohol] or an isoequivalent thereof, whereby to form a compound of formula VI as defined above or an isoequivalent thereof, (b) where X represents -O-P(O)(OR⁶ a )-O- and R⁶ a represents a hydrogen atom or a phosphate protecting group, reacting a phosphorylated biotinol derivative of the formula XIX set out hereinafter [wherein J, K, m and R⁶ a are as defined above] or an isoequivalent thereof, with an appropriate diol of formula IX, set out hereinafter [wherein n is as defined above], whereby to form a compound of formula VI or an isoequivalent thereof as defined above. (c) wherein X represents -S-, -O-, or N-R⁸, and R⁸ represents a straight chain or branched C₁₋₁₀ alkyl group, reacting a compound of formula X set out hereinafter [wherein J, K and m are as defined above and L′ represents a displaceable group] or an isoequivalent thereof, with a compound of formula XI set out hereinafter [wherein p is as defined above and Q represents the anion of a hydroxy group, a thiol group or an amine group of the formula N-R⁸ (wherein R⁸ is as defined above) whereby to form a compound of formula VI or an isoequivalent thereof as defined above. (d) with X represents -CONH-, which process comprises reacting a compound of the general formula XII set out hereinafter [wherein J, K and m are as defined above and R¹ represent a carboxylic acid group or a derivative thereof] or an isoequivalent thereof with a hydroxyalkylamine of the formula XIII set out hereinafter [wherein p is as defiend above] whereby to form a compound of formula VI or an isoequivalent thereof as defined above. (e) where X represents -NHCONH-, which process comprises reacting a compound of the general formula XVII set out hereinafter [wherein J, K and m are as defined above] or an isoequivalent thereof with a compound of the general formula XVIII set out hereinafter [wherein p is as defined above and W represents a displaceable group] whereby to form a compound of the formula VI or an isoequivalent thereof.

Claim 14

A compound of formula VI as set out hereinafter [in which J, K, m, p and X are as defined in any one of claims 1 to 8 above] or an isoequivalent thereof.

Claim 15

A compound of formula VI as claimed in claim 13 wherein neither J nor K may represent C₁₋₃alkyl, C₁₋₃alkenyl, acetyl, methoxy­carbonyl, phenyl, benzyl or benzoyl.

Claim 16

A compound of formula VII as set out hereinafter [in which J, K, m, p and X are as defined in any one of claims 1 to 8 above and R represents a group reducable to the corresponding alcohol, but neither J nor K may represent C₁₋₃alkyl, C₁₋₃alkenyl, acetyl, methoxycarbonyl, phenyl, benzyl or benzoyl] or an isoequivalent thereof.

Claim 17

A process for the preparation of a compound of the general formula VII, VIII or XII set out hereinafter [wherein J, K, m, p, R and R′ are as defined in claim 12 above] or an isoequivalent thereof, which process comprises reacting a corresponding derivative of the general formula XIV, XV or XVI set out hereinafter [wherein J, K, m, p, R and R′ are as defined above] or an isoequivalent thereof with a group of the formula J′ and/or with a group of the formula K′ [wherein the compounds J′ and K′ are precursors of the moieties J and K respectively as defined above] whereby to form a compound of formula VII, VIII or XII or an isoequivalent thereof as defined above.

Claim 18

A process which comprises two or more of the processes as claimed in any of claims 1 to 6, 8, 10, 12 and 16 the processes being carried out in sequence.