Non-Coding Immunomodulatory DNA Construct
The present invention relates to a nucleic acid molecule and its use for the modulation of the immune system. It provides a DNA construct for immunomodulation comprising at least one nucleotide in L-conformation.
1 . A DNA construct for immunomodulation comprising at least one sequence motif N 1 N 2 CGN 3 N 4 , wherein N is a nucleotide comprising is any A, C, T, or G, and C is deoxycytidine, G is deoxyguanosine, A is deoxyadenosine and T is deoxythymidine, and wherein at least one nucleotide is in L-conformation.
2 . The construct according to claim 1 , wherein N 1 N 2 is selected from the group consisting of GT, GG, GA, AT and AA, and N 3 N 4 is selected from the group consisting of CT and TT.
3 . The construct according to claim 1 , wherein at least one nucleotide in L-conformation is one of the five terminal nucleotides located at the 5′- and/or the 3′-end of a DNA single strand of the DNA construct.
4 . The construct according to claim 1 , wherein there is at least one G motif of at least three consecutive deoxyguanosines located near the 5′ and/or 3′ end of the DNA construct.
5 . The construct according to claim 1 , wherein a G motif of at least three consecutive deoxyguanosines is located between two of said sequence motifs N 1 N 2 CGN 3 N 4 .
6 . The construct according to claim 1 , wherein at least five nucleotides are located between two said sequences motifs N 1 N 2 CGN 3 N 4 , excluding deoxyguanosine.
7 . The construct according to claim 1 , wherein the DNA construct is a linear and open-chained comprising single or double-stranded DNA, or is linear, wherein said DNA construct comprises at least one end with a single stranded loop.
8 . The construct according to claim 1 , wherein the sequence motif N 1 N 2 CGN 3 N 4 is located within a single-stranded and/or a double-stranded region of the DNA sequence.
9 . The construct according to claim 1 , comprising inter- and/or intramolecular base-pairs and at least one unpaired, single-stranded region.
10 . The construct according to claim 21 , wherein at least two DNA constructs ligate with each other.
11 . The construct according to claim 1 , wherein at least one nucleotide in L- or D-conformation is modified with a functional group selected from the group consisting of carboxyl, amine, amide, aldimine, ketal, acetal, ester, ether, disulfide, thiol, and aldehyde.
12 . The construct according to claim 11 , wherein the modified nucleotide is linked to a compound selected from the group consisting of peptides, proteins, carbohydrates, antibodies, lipids, micelles, vesicles, synthetic molecules, polymers, micro projectiles, metal particles, nanoparticles, and a solid phase.
13 . The construct according to claim 1 , comprising a first G motif at the 5′ end and three sequence motifs N 1 N 2 CGN 3 N 4 , wherein at least five nucleotides are located between the first and second sequence motif N 1 N 2 CGN 3 N 4 , excluding deoxyguanosine, and a G motif, which is located between the second and third sequence motif N 1 N 2 CGN 3 N 4 , and wherein two of the three 3′ terminal deoxynucleotides are in L-conformation.
14 . The construct according to claim 13 , comprising at least one further G motif.
15 . A pharmaceutical composition comprising a DNA construct according to claim 1 .
16 . The pharmaceutical composition according to claim 15 , comprising additionally a chemotherapeutic.
17 . A vaccine comprising a DNA construct according to claim 1 .
18 . The vaccine according to claim 17 , wherein the DNA construct is an adjuvant.
19 . A method of treating cancer or an autoimmune disease, comprising administering the DNA construct according to claim 1 .
20 . A method of modulating the immune system, comprising administering the DNA construct according to claim 1 .
21 . The DNA construct according to claim 2 , wherein a G motif of at least three consecutive deoxyguanosines is located between two of said sequence motif N 1 N 2 CGN 3 N 4 .
22 . The DNA construct according to claim 2 , wherein at least five nucleotides are located between two said sequence motifs N 1 N 2 CGN 3 N 4 ′ excluding deoxyguanosine.
23 . The DNA construction according to claim 2 , comprising a first G motif at the 5′ end and three sequence motifs N 1 N 2 CGN 3 N 4 , wherein at least five nucleotides are located between the first and second sequence motif N 1 N 2 CGN 3 N 4 , excluding deoxyguanosine, and a G motif, which is located between the second and third sequence motif N 1 N 2 CGN 3 N 4 , and wherein two of the three 3′ terminal deoxynucleotides are in L-conformation.
24 . The DNA construct according to claim 23 , comprising at least one further G motif.