The present invention relates to new triphosphate-modified oligonucleotides which may act as RIG-I ligands as well as a new method allowing the synthesis and purification in high yield and purity suitable for pharmaceutical applications.
대표청구항▼
1. A modified oligonucleotide of formula (I) wherein V1, V3 and V5 are each independently selected from the group consisting of O, S and Se;V2, V4 and V6 are each independently selected from the group consisting of OH, OR1, SH, SR1, F, NH2, NHR1, N(R1)2 and BH3-M+,W1 is O or S,W2 is selected from th
1. A modified oligonucleotide of formula (I) wherein V1, V3 and V5 are each independently selected from the group consisting of O, S and Se;V2, V4 and V6 are each independently selected from the group consisting of OH, OR1, SH, SR1, F, NH2, NHR1, N(R1)2 and BH3-M+,W1 is O or S,W2 is selected from the group consisting of O, S, NH and NR2,W3 is selected from the group consisting of O, S, NH, NR2, CH2, CHHaI and C(HaI)2,R1, R2 and R3 are selected from the group consisting of H, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C2-6 acyl and a cyclic group, each substituted or unsubstituted,or wherein two R1 may form a ring together with an N-atom bound thereto,M+ is a cation,X is selected from the group consisting of NH, NR3, O and S,Z represents a capture tag, wherein the capture tag is selected from the group consisting of a long-chain aliphatic residue; Q or NHC2-C24 alkyl; a perfluoroalkyl entity; an azide or alkynyl group; a first partner of a non-covalent binding pair selected from the group consisting of biotin, desthiobiotin, a hapten, and an antigen; and a chemical entity containing a second amino group in an NH2—Y—XH type reagent; cholesterol; and a chloroformiate group, and wherein Q is selected from the group consisting of amino acids, C1-C24 alkyl, preferably C12-C24 alkyl, peptides and lipids,Y represents a bond or a linker connecting the capture tag to X, andON represents an oligonucleotide comprising at least 4 nucleotide or nucleotide analogue building blocks. 2. The modified oligonucleotide of claim 1, wherein V1, V2, V3, V4, V5, V6, W1, W2 and W3 are O. 3. The modified oligonucleotide of claim 1, wherein Y is either a bond or a linker which is selected from the group consisting of alkylenes, aralkylenes, and polyalkylene oxides. 4. The modified oligonucleotide of claim 1, wherein Z is selected from the group consisting of a long-chain aliphatic residue, Q and NHC2—C24 alkyl, and wherein Q is selected from the group consisting of amino acids, amino acid analogues, C1-C24 alkyl, peptides and lipids. 5. The modified oligonucleotide of claim 1, wherein the oligonucleotide is selected from the group consisting of single-stranded or double-stranded desoxyribonucleotides, ribonucleotides and oligonucleotide analogues, which are unmodified or chemically modified at the nucleoside and/or the ribose of the desoxyribonucleotide, ribonucleotide or oligonucleotide analogue. 6. The modified oligonucleotide of claim 1, comprising chemical modifications which maintain, establish or enhance the selectivity and/or the chemical stability of the oligonucleotide. 7. The modified oligonucleotide of claim 1, comprising chemical modifications independently selected from the group consisting of halogenation, 2′-O-alkylation, and at least one phosphorothioate modification between internukleotid linkages. 8. The modified oligonucleotide of claim 1, wherein X is NH or O,Y is —K—((CHR1)m—CH2—O)n—R, or—(O—(CHR3)m3—CH2)n1—(O—(CHR2)m2—CH2)n2—(O—(CHR1)m1—CH2)n3—,K is O or NH,m, m1, m2 and m3 are independently 1 to 12,n, n1, n2 and n3 are independently 0 to 20,R1, R2 and R3 are independently selected from the group consisting of H, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C2-C6-acyl and a cyclic group, each substituted or unsubstituted, andR is selected from the group consisting of C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C2-C6-acyl and a cyclic group, each substituted or unsubstituted. 9. The modified oligonucleotide of claim 8, wherein a) R1 and R2 are H, andn1=0, n2=1 and n3=1, orb) R1, R2 and R3 are H, andn1, n2 and n3 are each 1. 10. The modified oligonucleotide of claim 1, wherein X is NH or O,Y is a bond, andZ is a capture tag selected from the group consisting of C1-C12 alkyl, Q and NHC2—C24 alkyl, wherein Q is selected from the group consisting of amino acids, amino acid analogues, C1-C24 alkyl, peptides and lipids; andV1, V2, V3, V4, V5, V6, W1, W2 and W3 are O. 11. The modified oligonucleotide of claim 1, wherein X is O,Y is a bond,Z is H, andV1, V2, V3, V4, V5, V6, W1, W2 and W3 are O. 12. A pharmaceutical composition comprising an oligonucleotide according to claim 1 in combination with a pharmaceutically acceptable carrier, diluent and/or adjuvant. 13. The pharmaceutical composition of claim 12, wherein the pharmaceutically acceptable carrier, diluent and/or adjuvant is suitable for intradermal administration. 14. The pharmaceutical composition of claim 13, wherein the pharmaceutically acceptable carrier, diluent and/or adjuvant is suitable for intradermal administration by tattooing, microneedling or microneedle patches. 15. A method of preparing an oligonucleotide according to claim 1, comprising (a) reacting a compound of formula (IIa) wherein: V1, V3 and V5 are each independently selected from the group consisting of O, S and Se;V4 and V6 are each independently selected from the group consisting of OH, OR1, SH, SR1, F, NH2, NHR1, N(R1)2 and BH3-M+,W1 is O or S,W2 is selected from the group consisting of O, S, NH and NR2,W3 is selected from the group consisting of O, S, NH, NR2, CH2, CHHaI and C(HaI)2,R1 and R2 are selected from the group consisting of H, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C2-6 acyl and a cyclic group, each substituted or unsubstituted,or wherein two R1 may form a ring together with an N-atom bound thereto,M+ is a cation, andON represents an oligonucleotide comprising at least 4 nucleotide or nucleotide analogue building blocks, wherein ON is protected by at least one protection group,with an oxidizing agent to obtain a compound of formula (IIb) wherein V1, V3, V5, V4, V6, W1, W2, W3 and ON are as defined in claim 1, and V2 is selected from the group consisting of O, S and Se; wherein ON is protected by at least one protection group,(b) reacting a compound of formula (IIb) with a capture tag agent of formula (III), Z—Y—XH (III),whereinX is selected from the group consisting of NH, NR3, O and S,Z represents a capture tag, andY represents a bond or a linker connecting the capture tag to X, to obtain a reaction product comprising the oligonucleotide of formula (I), (c) deprotecting the at least one ON protection group, and(d) contacting the reaction product of step (b) with a capture reagent capable of interacting with the capture tag, wherein the contacting takes place under conditions which allow separation of the oligonucleotide (I) from other species contained in said reaction product; wherein the capture tag is selected from the group consisting of a long-chain aliphatic residue; Q or NHC2—C24 alkyl; a perfluoroalkyl entity; an azide or alkynyl group; a first partner of a non-covalent binding pair selected from biotin, desthiobiotin, a hapten, and an antigen, which has a binding constant of 10−6 I/mol or less with the capture reagent, which is a second complementary binding partner of the high-affinity binding pair; and a chemical entity containing a second amino group in an NH2-Y—XH type reagent; cholesterol; and a chloroformiate group, and wherein Q is selected from the group consisting of amino acids, C1-C24 alkyl, preferably C12-C24 alkyl, peptides and lipids; andwherein the capture reagent is a chromatographic material with affinity for hydrophobic or fluorinated groups; a second partner of a complementary binding partner of the high-affinity binding pair which is a streptavidin, an avidin, or an antibody; or an azide or alkynyl group. 16. The method according to claim 15, further comprising (e) removing the capture tag to obtain an oligonucleotide, wherein when X═O a compound of Formula (IVa) or (Ivb) is obtained and when X═NH a compound of Formula (IVa) is obtained. 17. The method according to claim 15, wherein X is O. 18. The modified oligonucleotide of claim 3, wherein said aralkylenes comprise heteroatoms or heteroatom-containing groups. 19. The modified oligonucleotide of claim 5, wherein the oligonucleotide is double stranded and each strand of the double strand has a length of at least 19 nucleotides. 20. The modified oligonucleotide of claim 7, wherein the chemical modifications are associated with the RIG-I selectivity of the oligonucleotide. 21. The modified oligonucleotide of claim 7, wherein the halogenation is F halogenation and the 2′-O-alkylation is 2′-O-methylation. 22. The modified oligonucleotide of claim 10, wherein Z is C10 or Q, wherein Q is C12-C24 alkyl. 23. A method for stimulating a patient's immune system, comprising administering a modified oligonucleotide of formula (I) wherein V1, V3 and V5 are each independently selected from the group consisting of O, S and Se;V2, V4 and V6 are each independently selected from the group consisting of OH, OR1, SH, SR1, F, NH2, NHR1, N(R1)2 and BH3-M+,W1 is O or S,W2 is selected from the group consisting of O, S, NH and NR2,W3 is selected from the group consisting of O, S, NH, NR2, CH2, CHHaI and C(HaI)2,R1, R2 and R3 are selected from the group consisting of H, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C2-6 acyl and a cyclic group, each substituted or unsubstituted,or wherein two R1 may form a ring together with an N-atom bound thereto,M+ is a cation,X is selected from the group consisting of NH, NR3, O and S,Z represents a capture tag, wherein the capture tag is selected from the group consisting of a long-chain aliphatic residue; Q or NHC2—C24 alkyl; a perfluoroalkyl entity; an azide or alkynyl group; a first partner of a non-covalent binding pair selected from biotin, desthiobiotin, a hapten, and an antigen; and a chemical entity containing a second amino group in an NH2—Y—XH type reagent; cholesterol; and a chloroformiate group, and wherein Q is selected from amino acids, C1-C24 alkyl, peptides and lipids,Y represents a bond or a linker connecting the capture tag to X, andON represents an oligonucleotide comprising at least 4 nucleotide or nucleotide analogue building blocks;to a patient in need of such stimulation. 24. The method according to claim 23, wherein said modified oligonucleotide selectively stimulates the RIG-1 signaling pathway. 25. The method according to claim 23, wherein said modified oligonucleotide is administered intraperitoneally, intramuscularly, intravenously, intranasally, subcutaneously, intradermally or intrathecally. 26. The method according to claim 25, wherein said modified oligonucleotide is administered intradermally by tattooing or by microneedling. 27. The method according to claim 23, wherein said C1-C24 alkyl is a C12-C24 alkyl.
연구과제 타임라인
LOADING...
LOADING...
LOADING...
LOADING...
LOADING...
이 특허에 인용된 특허 (36)
Ecker David J. (Carlsbad CA), Antisense inhibitors of the human immunodeficiency virus phosphorothioate oligonucleotides.
Wilfried Seifert, Compositions and methods for inhibiting cox-2 expression and treating cox-2 associated disorders by using cox-2 antisense oligonucleotides.
Agrawal Sudhir (Shrewsbury MA) Leiter Josef M. E. (Brooklyn NY) Palese Peter (Leonia NJ) Zamecnik Paul C. (Shrewsbury MA), Inhibition of influenza virus replication by oligonucleotide phosphorothioates.
Harel Bellan,Annick; Ait Si Ali,Slimane; Cabon Georget,Florence; Chauchereau,Anne; Dautry,Francois, Inhibitor oligonucleotides and their use for specific repression of a gene.
Cohen Jack S. (Bethesda MD) Neckers Len (Bethesda MD) Stein Cy (Gaithersburg MD) Loke She L. (Wheaton MD) Shinozuka Kazuo (Kazo JPX), Inhibitors for replication of retroviruses and for the expression of oncogene products.
Stec Wojciech J. (Lodz PLX) Grajkowski Andrzej (Lodz PLX) Uznanski Bogdan (Lodz PLX), Method of making oligonucleotides and oligonucleotide analogs using phospholanes and enantiomerically resolved phosphola.
Pederson Thoru (Worcester MA) Agrawal Sudhir (Shrewsbury MA) Mayrand Sandra (Shrewsbury MA) Zamecnik Paul C. (Shrewsbury MA), Method of site-specific alteration of RNA and production of encoded polypeptides.
Pederson Thoru (Worcester MA) Agrawal Sudhir (Shrewsbury MA) Mayrand Sandra (Shrewsbury MA) Zamecnik Paul C. (Shrewsbury MA), Method of site-specific alteration of RNA and production of encoded polypeptides.
Stec Wojciech J. (Lodz PLX) Uznanski Bogdan (Lodz CA PLX) Bergot B. John (Redwood City CA) Hirschbein Bernard L. (San Francisco CA) Fearson Karen L. (Union City CA), Method of synthesizing sulfurized oligonucleotide analogs.
Cook, Phillip D.; Wang, Guangyi; Bruice, Thomas W.; Boyle, Nicholas A.; Leeds, Janet M.; Brooks, Jennifer L.; Prhavc, Marija; Ariza, Maria Eugenia; Fagan, Patrick C.; Jin, Yi; Rajwanshi, Vivek K.; Tucker, Kathleen D., Nucleotide mimics and their prodrugs.
Cook, Phillip D.; Wang, Guangyi; Bruice, Thomas W.; Boyle, Nicholas A.; Leeds, Janet M.; Brooks, Jennifer L.; Prhavc, Marija; Ariza, Maria Eugenia; Fagan, Patrick C.; Jin, Yi; Rajwanshi, Vivek K.; Tucker, Kathleen D., Nucleotide mimics and their prodrugs.
Eppstein Deborah A. (Los Altos CA) Fraser-Smith Elizabeth B. (Los Altos CA) Matthews Thomas R. (Los Gatos CA), Serial injection of muramyldipeptides and liposomes enhances the anti-infective activity of muramyldipeptides.
※ AI-Helper는 부적절한 답변을 할 수 있습니다.