Polyamide nucleic acid derivatives, and agents, and processes for preparing them

1. A process for preparing a PNA derivative of Formula I

wherein

V is oxygen, sulfur, NR 1 , U—(CR 3 R 4 ) u ′—CH 2 —C(O)—NH or U—(CH 2 CH 2 O) u ′—C(O)—NH;

U is, independently of any other U, oxygen, sulfur or NH;

u′ is, independently of any other u′, from 1 to 10;

W is, independently of any other W, oxygen, sulfur or NR 1 ;

Y is, independently of any other Y, hydroxyl, mercapto, oxyanion, thioate or NR 1 R 2 ;

R 1 and R 2 are, independently of each other, a radical consisting of hydrogen or C 1 -C 6 -alkyl;

R 3 and R 4 are, independently of each other, a radical consisting of hydrogen or C 1 -C 6 -alkyl, or the radical of an amino acid side chain;

X is, independently of any other X,

U—(C 2 -C 22 -alkanediyl)-U,

U—(CH 2 CH 2 O) u ′,

a bifunctional labeling group,

a bifunctional group for crosslinking with complementary nucleic acids,

a bifunctional group which promotes intracellular uptake, or

a bifunctional group which increases the binding affinity of the PNA derivative for a

target nucleic acid;

Z is hydroxyl,

mercapto,

oxyanion,

thioate,

NR 1 R 2 ,

C 1 -C 22 -alkyl,

C 1 -C 8 -arylalkyl,

C 1 -C 22 -alkyl-U,

C 1 -C 8 -arylalkyl-U,

hydroxy-C 1 -C 18 -U,

aminoalkyl-U,

arylalkyl-U,

mercaptoalkyl-U,

a group of the formula R 9 (CH 2 CH 2 -O) m , wherein R 9 is hydroxyl, amino or C 1 -C 22 -alkoxy,

and m is from 1 to 100,

a monofunctional or bifunctional labeling group,

a monofunctional or bifunctional crosslinking group,

a monofunctional or bifunctional group which promotes intracellular uptake, or

a monofunctional or bifunctional group which promotes the binding affinity of the

PNA derivative for a target nucleic acid;

n is from 0 to 10;

Q is hydroxyl, amino, NHR 7 , NR 7 R 8 , an amino acid derivative or a peptide radical;

R 7 and R 8 are, independently of each other, C 1 -C 18 -alkyl or hydroxy-C 1 -C 18 -alkyl;

and wherein {POLY} is described by the Formula II

wherein {BLOCK} is, independently of any other {BLOCK}, described by Formula IIIA,

wherein each building block {BLOCK} can be different,

and wherein

z′ is from 0 to 100;

A is, independently of any other A, (CR 1 R 2 ) s , wherein s is from 1 to 3;

B is, independently of any other B,

an aromatic radical, which optionally possesses heteroaromatic character,

hydrogen,

hydroxyl,

C 1 -C 18 -alkyl,

or a nucleobase or its prodrug form,

wherein at least one B radical is a nucleobase;

D is, independently of any other D, (CR 3 R 4 ) t , wherein t is from 2 to 10,

wherein two adjacent R 3 and R 4 radicals can form a C 5 -C 8 -cycloalkyl ring,

E is, independently of any other E, (CR 5 R 6 ) u ′,

R 5 and R 6 are, independently of each other, a radical consisting of hydrogen or C 1 -C 6 -alkyl,

or an amino acid side chain, wherein two adjacent R 5 and R 6 radicals

can form a C 5 -C 8 -cycloalkyl ring or a spiro compound, and

wherein R 1 , R 2 , R 3 , R 4 and u′ are as defined above;

and physiologically tolerated salts of the PNA derivative of the Formula I,

with the proviso that at least one Y or Z radical is hydroxyl, mercapto, oxyanion or thioate;

wherein said process comprises

a) synthesizing a backbone for the PNA derivative, starting from the C terminus, by sequentially coupling with amidonucleic acid monomes, which are optionally N-terminally protected;

b) optionally deprotecting the N-terminally protected PNA backbone;

c) coupling a phosphorus (III) or a phosphorus (V) group to the N-terminus of the PNA backbone, using activated phosphorylating reagents optionally containing a spacer;

d) optionally repeating step c); and

e) optionally oxidizing the phosphorus (III) group to a phosphorus (V) group.

2. The process as claimed in claim 1 , wherein the PNA is prepared using t-butyloxycarbonyl (BOG), 9-fluorenylmethoxycarbonyl (Fmoc), or monomethoxytrityl (Mmt) protecting groups.

3. The process as claimed in claim 2 , wherein the PNA is prepared using solid supports.

4. The process as claimed in claim 3 , wherein CPG, tentagel, or aminomethylpolystyrene is used as the solid support.

5. The process as claimed in claim 1 , further comprising purifying the PNA derivative using chromatography or electrophoresis.

6. The process as claimed in claim 5 , wherein the PNA derivative is purified using chromatography using a basic stationary phase and an acid or salt-containing eluent.

7. The process as claimed in claim 6 , wherein the stationary phase is an anion exchanger or a mixed-mode phase.