US2010178272A1PendingUtilityA1
Structure and use of 5'phosphate oligonucleotides
Est. expiryAug 8, 2026(~0 yrs left)· nominal 20-yr term from priority
A61P 37/00A61P 43/00A61P 37/08A61P 37/04A61P 37/02A61P 37/06A61P 31/12A61P 35/00A61P 31/04A61P 25/00A61P 33/00A61P 31/00C12N 2310/351A61K 45/06C12N 2310/335C07H 21/00C12N 2310/17C12N 2310/31A61K 31/7115A61K 31/7105C12N 15/117C12N 2310/32C12N 2320/30C12N 2310/346Y02A50/30
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Claims
Abstract
Oligonucleotides bearing free, uncapped 5′ phosphate group(s) are recognized by RIG-I, leading to the induction of type I IFN, IL-18 and IL-1β production. Bacterial RNA also induces type I IFN production. 5′ phosphate oligonucleotides and bacterial RNA can be used for inducing an anti-viral response or an anti-bacterial response, in particular, type I IFN and/or IL-18 and/or IL-1β production, in vitro and in vivo and for treating various disorders and diseases such as viral infections, bacterial infections, parasitic infections, tumors, allergies, autoimmune diseases, immunodeficiencies and immunosuppression. Single-stranded 5′ triphosphate RNA can be used for inducing an anti-viral response, an anti-bacterial response, or an anti-tumor response, in particular, type I IFN and/or IL-18 and/or IL-1β production, in a target cell-specific manner.
Claims
exact text as granted — not AI-modified1 . An oligonucleotide or a precursor thereof for preventing and/or treating a disease and/or disorder selected from the group consisting of viral infection, bacterial infection, parasitic infection, tumor, multiple sclerosis, allergy, autoimmune diseases, immunosuppression and immunodeficiency in a vertebrate animal,
wherein the oligonucleotide comprises at least 1, preferably at least 3, more preferably at least 6 ribonucleotide(s) at the 5′ end, wherein the oligonucleotide comprises at least 1, preferably at least 2, and more preferably at least 3 phosphate group(s) at the 5′ end, and wherein the phosphate group is free of any cap or modification, wherein the oligonucleotide is at least 12, preferably at least 18, more preferably at least 20, and even more preferably at least 21 nucleotides in length.
2 . The oligonucleotide or the precursor thereof of claim 1 , wherein the oligonucleotide is single-stranded which is free of any sequence that is capable of forming a double-stranded structure, and wherein the nucleotide sequence of the oligonucleotide is complementary to a disease or disorder-related RNA,
3 . The oligonucleotide or the precursor thereof of claim 1 or 2 , wherein at least one of the 5′ uncapped phosphate groups is not comprised in a triphosphate.
4 . The oligonucleotide or the precursor thereof of any of claims 1 - 3 , wherein the oligonucleotide comprises at least one inosine (I).
5 . The oligonucleotide or the precursor thereof of any of claims 1 - 4 , wherein the first ribonucleotide at the 5′ end of the oligonucleotide comprises a ribonucleotide selected from A, C, U, preferably a ribonucletoide selected from A and C, and most preferably A.
6 . The oligonucleotide or the precursor thereof of any of claims 1 - 5 , wherein the sequence of the first 4 nucleotides at the 5′ end of the oligonucleotide is selected from: AAGU, AAAG, AUGG, AUUA, AACG, AUGA, AGUU, AUUG, AACA, AGAA, AGCA, AACU, AUCG, AGGA, AUCA, AUGC, AGUA, AAGC, AACC, AGGU, AAAC, AUGU, ACUG, ACGA, ACAG, AAGG, ACAU, ACGC, AAAU, ACGG, AUUC, AGUG, ACAA, AUCC, AGUC, wherein the sequence is in the 5′->3′ direction.
7 . The oligonucleotide or the precursor thereof of any of claims 1 - 6 , wherein the oligonucleotide is free of modifications such as pseudouridine, 2-thiouridine, 2′-Fluorine-dNTP, 2′-O-methylated NTP, in particular 2′-fluorine-dCTP, 2′-fluorine-dUTP, 2′-O-methylated CTP, 2′-O-methylated UTP.
8 . The oligonucleotide or the precursor thereof of any of claims 1 - 7 , wherein the oligonucleotide or the precursor thereof comprises at least one, preferably at least two, more preferably at least three, even more preferably at least four, even more preferably at least five, and most preferably at least six, of the 4-nucleotide (4 mer) motifs selected from the group consisting of:
GUUC,
(No. 1)
GUCA,
(No. 2)
GCUC,
(No. 3)
GUUG,
(No. 4)
GUUU,
(No. 5)
GGUU,
(No. 6)
GUGU,
(No. 7)
GGUC,
(No. 8)
GUCU,
(No. 9)
GUCC,
(No. 10)
GCUU,
(No. 11)
UUGU,
(No. 12)
UGUC,
(No. 13)
CUGU,
(No. 14)
CGUC,
(No. 15)
UGUU,
(No. 16)
GUUA,
(No. 17)
UGUA,
(No. 18)
UUUC,
(No. 19)
UGUG,
(No. 20)
GGUA,
(No. 21)
GUCG,
(No. 22)
UUUG,
(No. 23)
UGGU,
(No. 24)
GUGG,
(No. 25)
GUGC,
(No. 26)
GUAC,
(No. 27)
GUAU,
(No. 28)
UAGU,
(No. 29)
GUAG,
(No. 30)
UUCA,
(No. 31)
UUGG,
(No. 32)
UCUC,
(No. 33)
CAGU,
(No. 34)
UUCG,
(No. 35)
CUUC,
(No. 36)
GAGU,
(No. 37)
GGUG,
(No. 38)
UUGC,
(No. 39)
UUUU,
(No. 40)
CUCA,
(No. 41)
UCGU,
(No. 42)
UUCU,
(No. 43)
UGGC,
(No. 44)
CGUU,
(No. 45)
CUUG,
(No. 46)
UUAC,
(No. 47)
wherein the nucleotide sequences of the motifs are 5′→3′, and
wherein the oligonucleotide is between 12 and 64, preferably between 12 and 50, more preferably between 14 and 40, even more preferably between 16 and 36, and most preferably between 18 and 25 nucleotides in length.
9 . The oligonucleotide or the precursor thereof of any of claims 1 - 8 , wherein the oligonucleotide or the precursor thereof comprises at least one, preferably at least two, more preferably at least three, even more preferably at least four, even more preferably at least five, and most preferably at least six, of the 4-nucleotide (4 mer) motifs selected from the group consisting of:
UCGU,
(No. 1)
GUUG,
(No. 2)
UGGU,
(No. 3)
UGGC,
(No. 4)
GGUA,
(No. 5)
UGAU,
(No. 6)
UGCU,
(No. 7)
UUGC,
(No. 8)
UUGU,
(No. 9)
UAGU,
(No. 10)
GGUU,
(No. 11)
GUUU,
(No. 12)
UGUG,
(No. 13)
GUGU,
(No. 14)
UGCC,
(No. 15)
GUAU,
(No. 16)
GUGC,
(No. 17)
UGUA,
(No. 18)
UGUC,
(No. 19)
CUGU,
(No. 20)
UGAC,
(No. 21)
UGUU,
(No. 22)
UAAU,
(No. 23)
GUAG,
(No. 24)
UCUU,
(No. 25)
UUGG,
(No. 26)
UUUG,
(No. 27)
GGAU,
(No. 28)
UUUU,
(No. 29)
CGUU,
(No. 30)
UUAU,
(No. 31)
GUUC,
(No. 32)
GUGG,
(No. 33)
GGUG,
(No. 34)
UAUU,
(No. 35)
UCUG,
(No. 36)
GUAC,
(No. 37)
UAGG,
(No. 38)
UCUC,
(No. 39)
UAGC,
(No. 40)
UAUC,
(No. 41)
CUAU,
(No. 42)
UACU,
(No. 43)
CGGU,
(No. 44)
UGCG,
(No. 45)
UUUC,
(No. 46)
UAUG,
(No. 47)
UAAG,
(No. 48)
UACC,
(No. 49)
UUAG,
(No. 50)
GCUU,
(No. 51)
CAGU,
(No. 52)
UGAG,
(No. 53)
GAUU,
(No. 54)
GAGU,
(No. 55)
GUUA,
(No. 56)
UGCA,
(No. 57)
UUCU,
(No. 58)
GCCU,
(No. 59)
GGUC,
(No. 60)
GGCU,
(No. 61)
UUAC,
(No. 62)
UCAU,
(No. 63)
GCGU,
(No. 64)
GCAU,
(No. 65)
GAUG,
(No. 66)
GUCU,
(No. 67)
CGUA,
(No. 68)
CGAU,
(No. 69)
wherein the nucleotide sequences of the motifs are 5′→3′, and
wherein the oligonucleotide or precursor thereof is between 12 and 64, preferably between 12 and 50, more preferably between 14 and 40, even more preferably between 16 and 36, and most preferably between 18 and 30 nucleotides in length.
10 . The oligonucleotide or the precursor thereof of any of claims 1 - 9 , further comprising a complexation agent.
11 . The oligonucleotide or the precursor thereof of any of claims 1 - 9 , wherein the oligonucleotide or the precursor thereof is comprised in a viral vector.
12 . The oligonucleotide or the precursor thereof of any of claims 1 - 11 , wherein the oligonucleotide or the precursor thereof is for use in combination with at least one agent selected from an immunostimulatory agent, an anti-viral agent, an anti-bacterial agent, an anti-tumor agent, and a gene silencing agent, and/or an anti-tumor therapy.
13 . The oligonucleotide or the precursor thereof of claim 12 , wherein the oligonucleotide or the precursor thereof is for use in combination with retinoic acid and/or type I IFN.
14 . An oligonucleotide or a precursor thereof as defined in any of claims 1 - 13 for inducing apoptosis of tumor cells, inducing an anti-viral response, inducing an anti-bacterial response, and/or inducing an anti-tumor response in a vertebrate animal.
15 . The oligonucleotide or a precursor thereof of claim 14 , wherein the anti-viral response, the anti-bacterial response and/or the anti-tumor response comprise type I IFN production, IL-18 production, and/or IL-1β production.
16 . An oligonucleotide or a precursor thereof as defined in any of claims 1 - 13 and at least one antigen for inducing an immune response against the at least one antigen in a vertebrate animal.
17 . The oligonucleotide or the precursor thereof and the at least one antigen of claim 16 , wherein the oligonucleotide or the precursor thereof is covalently linked to the at least one antigen.
18 . Use of an oligonucleotide or a precursor thereof as defined in any of claims 1 - 13 for the preparation of a medicament for preventing and/or treating a disease and/or disorder selected from the group consisting of viral infection, bacterial infection, parasitic infection, tumor, multiple sclerosis, allergy, autoimmune diseases, immunosuppression and immunodeficiency in a vertebrate animal.
19 . Use of a oligonucleotide or a precursor thereof as defined in any of claims 1 - 13 for the preparation of a medicament for inducing apoptosis of tumor cells, inducing an anti-viral response, inducing an anti-bacterial response and/or inducing an anti-tumor response in a vertebrate animal.
20 . The use of claim 19 , wherein the anti-viral response, the anti-bacterial response, and/or the anti-tumor response comprise type I IFN production, IL-18 production, and/or IL-1β production.
21 . A pharmaceutical composition comprising a oligonucleotide or a precursor thereof as defined in any of claims 1 - 13 .
22 . A combined preparation comprising an oligonucleotide or a precursor thereof as defined in any of claims 1 - 13 and at least one agent selected from an immunostimulatory agent, an anti-viral agent, an anti-bacterial agent, an anti-tumor agent, and a gene silencing agent, wherein the oligonucleotide or the precursor thereof and the at least one agent are for simultaneous, separate or sequential administration.
23 . The combined preparation of claim 22 , wherein the agent is at least one of retinoic acid and type I IFN.
24 . A pharmaceutical package comprising the pharmaceutical composition of claim 21 or the combined preparation of claim 22 or 23 and an instruction for use.
25 . An in vitro method for stimulating an anti-viral and/or an anti-bacterial and/or an anti-tumor response in a cell, comprising the steps of:
(a) mixing an oligonucleotide or precursor thereof as defined in any of claims 1 - 9 with a complexation agent; and (b) contacting a cell with the mixture of (a), wherein the cell expresses RIG-I and/or components of the inflammasome.
26 . The method of claim 25 , wherein the oligonucleotide is single-stranded and wherein the cell contains a mRNA comprising a nucleotide sequence which is complementary to the nucleotide sequence of the oligonucleotide.
27 . A method for preparing an oligonucleotide capable of inducing an anti-viral and/or an anti-bacterial and/or anti-tumor response, comprising the steps of:
(a) introducing at least one uncapped 5′ monophosphate, diphosphate and/or triphosphate into an oligonucleotide; and (b) introducing a nucleotide sequence capable of forming double-stranded structure inside a cell into the oligonucleotide.
28 . The method claim 27 , further comprising the step(s) of:
(c) preparing an oligonucleotide with adenosine (A) at the 5′ end; and/or (d) preparing an olignucletide having a sequence selected from AAGU, AAAG, AUGG, AUUA, AACG, AUGA, AGUU, AUUG, AACA, AGAA, AGCA, AACU, AUCG, AGGA, AUCA, AUGC, AGUA, AAGC, AACC, AGGU, AAAC, AUGU, ACUG, ACGA, ACAG, AAGG, ACAU, ACGC, AAAU, ACGG, AUUC, AGUG, ACAA, AUCC, AGUC at the 5′ end; and/or (e) introducing inosine (I) into the oligonucleotide.
29 . A method for preparing an oligonucleotide free of anti-viral response-inducing activity and anti-bacterial response-inducing activity, comprising the step(s) of:
(a) eliminating all 5′ phosphate groups from the oligonucleotide; and/or (b) capping all 5′ monophosphate, diphosphate and triphosphate of the oligonucleotide; and/or (c) eliminating any nucleotide sequence capable of forming double-stranded structure inside a cell from the oligonucleotide; and/or (d) incorporating modified nucleotides such as pseudouridine, 2-thiouridine, 2′-Fluorine-dNTPs-2′-O-methylated NTPs, preferably 2′-fluorine-dCTP, 2′-fluorine-dUTP, 2′-O-methylated CTP, 2′-O-methylated UTP, into the oligonucleotide.
30 . The method of any of claims claim 25 - 29 , wherein the anti-viral response, the anti-bacterial response, and/or the anti-tumor response comprise type I IFN production, IL-18 production, and/or IL-1β production.
31 . A bacterial RNA for preventing and/or treating a disease and/or disorder selected from the group consisting of viral infection, bacterial infection, parasitic infection, tumor, multiple sclerosis, allergy, autoimmune diseases, immunosuppression and immunodeficiency in a vertebrate animal.Join the waitlist — get patent alerts
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