US2021196831A1PendingUtilityA1
Oligonucleotide constructs and uses thereof
Est. expiryNov 2, 2037(~11.3 yrs left)· nominal 20-yr term from priority
A61P 31/20A61P 35/00A61K 2039/55561A61K 2039/58A61K 47/545A61K 31/7088A61K 47/26A61K 9/0019A61K 2039/82A61K 47/14A61K 39/39A61K 47/22C12N 2730/10134A61K 47/543A61K 2039/6025C12Q 1/6876C12N 15/11A61K 2039/585A61K 2039/6018A61K 47/554A61K 9/19
55
PatentIndex Score
0
Cited by
0
References
0
Claims
Abstract
Oligodeoxynucleotide-based immunostimulatory Toll-Like Receptor 9 (TLR9) agonists are described. Also described are compositions comprising the TLR9 agonists, methods of making the TLR9 agonists, and methods of using the TLR9 agonists to treat immune diseases, disorders or conditions, such as viral infections or cancer.
Claims
exact text as granted — not AI-modified1 . A CpG ODN construct having a formula of:
5′M-L-[CpG ODN]3′, or Formula (IIa):
5′[CpG ODN]-L-M3′ Formula (IIb):
wherein:
M represents a lipid moiety, preferably the lipid moiety comprises at least one lipid moiety selected from the group consisting of cholesterol, tocopherol, a palmitoyl group, and a stearyl group;
L represents a linker comprising 10-100 atoms selected from the group consisting of carbon, nitrogen, oxygen, hydrogen, sulfur and phosphorus, wherein L is covalently linked to the CpG ODN via a cleavable linkage, preferably via an ester or an amide bond; and
CpG ODN comprises at least one CpG motif having a po internucleotide linkage;
optionally, the CpG ODN construct of Formula (IIa) or (IIb) is further covalently conjugated to a targeting moiety, which is preferably selected from the group consisting of galactose, N-acetylgalactosamine (GalNAc), fucose, mannose, sialic acid, N-acetyl neuraminic acid.
2 . A CpG ODN construct having a structure of:
5′M 1 -Y 1 —(((CH 2 ) 2 O) m —X) n —Y 2 -[CpG ODN]-Y 3 -M 2 3′, or Formula (IIIa):
5′M 2 -Y 3 -[CpG ODN]-Y 2 —(((CH 2 ) 2 O) m —X) n —Y 1 -M 1 3′ Formula (IIIb):
wherein: M 1 represents a lipid moiety, preferably the lipid moiety comprises at least one selected from the group consisting of cholesterol, tocopherol, a palmitoyl group, and a stearyl group, M 2 represents a lipid moiety, a targeting moiety, or is absent, Y 1 is a bond or a linker, preferably ethylene glycol having the formula ((CH 2 ) 2 O) o , wherein o is 1-15, covalently linked to (((CH 2 ) 2 O) m —X) n via a phosphodiester (po) linkage, a phosphorothioate (ps) linkage or a bond, X is independently a bond, a po linkage or a ps linkage, each of Y 2 and Y 3 is independently a cleavable linkage, preferably comprises an ester or an amide bond, more preferably comprises a po linkage or a phosphoramidate linkage, most preferably a po linkage, provided that when M 2 is absent, Y 3 is absent, CpG ODN comprises at least one CpG motif having a po internucleotide linkage, preferably comprises at least two, three or four CpG motifs each having the po internucleotide linkage; m is an integer from 1 to 15, and n is an integer from 1 to 5.
3 . The ODN construct of claim 2 , having a formula of:
5′M-Y 1 —(((CH 2 ) 2 O) m —X) n —Y 2 -[CpG ODN]3′, or Formula (IVa):
5′[CpG ODN]-Y 2 —(((CH 2 ) 2 O) m —X) n —Y 1 -M3′ Formula (IVb):
wherein: M represents a lipid moiety, preferably the lipid moiety comprises at least one lipid moiety selected from the group consisting of cholesterol, tocopherol, a palmitoyl group, and a stearyl group, Y 1 is a bond or a linker, preferably ethylene glycol having the formula ((CH 2 ) 2 O) o , wherein o is 1-15, covalently linked to (((CH 2 ) 2 O) m —X) n via a phosphodiester (po) linkage, a phosphorothioate (ps) linkage or a bond, X is independently a bond, a phosphodiester (po) linkage or a phosphorothioate (ps) linkage, Y 2 is a cleavable linkage, preferably comprises an ester or an amide bond, more preferably comprises a po linkage or a phosphoramidate linkage, most preferably a po linkage; CpG ODN comprises at least one CpG motif having a po internucleotide linkage, preferably comprises at least two, three or four CpG motifs each having the po internucleotide linkage; m is an integer from 1 to 15, preferably 6; and n is an integer from 1 to 5, preferably 2; optionally, the CpG ODN construct of Formula (IVa) or (IVb) is further covalently conjugated to a targeting moiety, which is preferably selected from the group consisting of galactose, N-acetylgalactosamine (GalNAc), fucose, mannose, sialic acid, N-acetyl neuraminic acid.
4 . The ODN construct of claim 3 , having a formula construct selected from the group consisting of:
5′M-po(HEG)po(HEG)po-[CpG ODN]3′;
5′M-ps(HEG)po(HEG)po-[CpG ODN]3′,
5′M-ps(HEG)ps(HEG)po-[CpG ODN]3′,
5′M-po(HEG)ps(HEG)po-[CpG ODN]3′,
5′[CpG ODN]-po(HEG)po(HEG)po-M3′,
5′[CpG ODN]-po(HEG)ps(HEG)po-M3′,
5′[CpG ODN]-po(HEG)ps(HEG)ps-M3′, and
5′[CpG ODN]-po(HEG)po(HEG)ps-M3′,
wherein:
M represents a lipid moiety comprising at least one selected from the group consisting of cholesterol, tocopherol, a palmitoyl group, and a stearyl group;
po represents a phosphodiester linkage;
HEG represents ((CH 2 ) 2 O) 6 ;
ps represents a phosphorothioate linkage; and
CpG ODN comprises at least one CpG motif having a po internucleotide linkage, preferably comprises at least two, three or four CpG motifs each having the po internucleotide linkage,
wherein M is covalently linked to (HEG) directly via a po or ps linkage, or indirectly via a second linker that is linked to (HEG) directly via a po or ps linkage.
5 . The ODN construct of claim 3 , having a formula selected from the group consisting of:
5′Toco-po(HEG)po(HEG)po-[CpG ODN]3′, wherein Toco represents tocopherol,
5′Chol-po(HEG)po(HEG)po-[CpG ODN]3′, wherein Chol represents cholesterol,
5′Palm-po(HEG)po(HEG)po-[CpG ODN]3′, wherein Palm represents a palmitoyl group,
5′[CpG ODN]-po(HEG)ps-Chol3′, wherein Chol represents cholesterol,
5′[CpG ODN]-po(HEG)ps-Toco3′, wherein Toco represents tocopherol, and
5′[CpG ODN]-po(HEG)ps-Palm3′, wherein Palm represents a palmitoyl group,
wherein:
po represents a phosphodiester linkage;
HEG represents ((CH 2 ) 2 O) 6 ;
ps represents a phosphorothioate linkage; and
CpG ODN comprises at least one CpG motif having a po internucleotide linkage, preferably comprises at least two, three or four CpG motifs each having the po internucleotide linkage,
wherein the tocopherol, the cholesterol, or the palmitoyl group is covalently linked to the (HEG) directly via a po or ps linkage, or indirectly via a second linker that is linked to (HEG) directly via a po or ps linkage.
6 . The ODN construct of claim 1 , wherein the CpG ODN has a phosphorothioate (ps) internucleotide linkage.
7 . The ODN construct of claim 1 , wherein two to four of the CpG dinucleotides in the CpG ODN each have a phosphodiester (po) internucleotide linkage.
8 . The ODN construct of claim 1 , wherein the CpG ODN comprises a polynucleotide sequence selected from the group consisting of:
(1)
(SEQ ID NO: 1)
5′ TCGTCGTTTTGTCGTTTTGTCGTT 3′;
(2)
(SEQ ID NO: 2)
5′ GGGGGACGATCGTCGGGGGG 3′;
(3)
(SEQ ID NO: 3)
5′ GGGGTCAACGTTGAGGGGGG 3′;
(4)
(SEQ ID NO: 4)
5′ TCCATGACGTTCCTGACGTT 3′;
(5)
(SEQ ID NO: 5)
5′ TCGTCGTTTTCGGCGCGCGCCG 3′;
(6)
(SEQ ID NO: 6)
5′ TCGTCGTTACGTAACGACGACGTT 3′;
and
(7)
(SEQ ID NO: 7)
5′ TCGTCGTTTTGTCGTTTTGTCGT 3′.
9 . The ODN construct of claim 1 , wherein the CpG ODN comprises a polynucleotide sequence selected from the group consisting of:
(1)
(SEQ ID NO: 8)
5' TpsCpoGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTps
TpsTpsTpsGpsTpsCpoGpsTpsT 3';
(2)
(SEQ ID NO: 9)
5' TpsCpsGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpsGpsTps
TpsTpsTpsGpsTpsCpsGpsTpsT 3';
(3)
(SEQ ID NO: 10)
5' TpsCpsGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTps
TpsTpsTpsGpsTpsCpsGpsTpsT 3';
(4)
(SEQ ID NO: 11)
5' TpsCpsGpsTpsCpsGpsTpsTpsTpsTpsGpsTpsCpoGpsTps
TpsTpsTpsGpsTpsCpsGpsTpsT 3';
(5)
(SEQ ID NO: 12)
5' TpsCpsGpsTpsCpsGpsTpsTpsTpsTpsGpsTpsCpsGpsTps
TpsTpsTpsGpsTpsCpoGpsTpsT 3';
(6)
(SEQ ID NO: 13)
5' TpsCpsGpsTpsCpsGpsTpsTpsTpsTpsGpsTpsCpoGpsTps
TpsTpsTpsGpsTpsCpoGpsTpsT 3';
(7)
(SEQ ID NO: 14)
5' TpsCpsGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTps
TpsTpsTpsGpsTpsCpoGpsTpsT 3';
(8)
(SEQ ID NO: 15)
5' TpsCpsGpsTpsCpsGpsTpsTpsApsCpsGpsTpsApsApsCps
GpsApsCpsGpsApsCpsGpsTpsT 3';
(9)
(SEQ ID NO: 16)
5' TpsCpsGpsTpsCpoGpsTpsTpsApsCpsGpsTpsApsApsCps
GpsApsCpsGpsApsCpsGpsTpsT 3';
(10)
(SEQ ID NO: 17)
5' TpsCpsGpsTpsCpoGpsTpsTpsApsCpoGpsTpsApsApsCps
GpsApsCpsGpsApsCpsGpsTpsT 3';
(11)
(SEQ ID NO: 18)
5' TpsCpsGpsTpsCpoGpsTpsTpsApsCpoGpsTpsApsApsCpo
GpsApsCpsGpsApsCpsGpsTpsT 3';
(12)
(SEQ ID NO: 19)
5' TpsCpsGpsTpsCpoGpsTpsTpsApsCpoGpsTpsApsApsCpo
GpsApsCpoGpsApsCpsGpsTpsT 3';
(13)
(SEQ ID NO: 20)
5' GpsGpsGpsGpsGpsApsCpoGpsApsTpsCpoGpsTpsCpoGps
GpsGpsGpsGpsG 3';
(14)
(SEQ ID NO: 21)
5' GpsGpsGpsGpsTpsCpsApsApsCpoGpsTpsTpsGpsApsGps
GpsGpsGpsGpsG 3';
(15)
(SEQ ID NO: 22)
5' TpsCpsCpsApsTpsGpsApsCpsGpsTpsTpsCpsCpsTpsGps
ApsCpsGpsTpsT 3';
(16)
(SEQ ID NO: 23)
5' TpsCpsGpsTpsCpsGpsTpsTpsTpsTpsCpsGpsGpsCpsGps
CpsGpsCpsGpsCpsCpsG 3';
(17)
(SEQ ID NO: 24)
5' TpsCpoGpsTpsCpsGpsTpsTpsTpsTpsGpsTpsCpsGpsTps
TpsTpsTpsGpsTpsCpsGpsT 3';
(18)
(SEQ ID NO: 25)
5' TpsCpoGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpsGpsTps
TpsTpsTpsGpsTpsCpsGpsT 3';
and
(19)
(SEQ ID NO: 26)
5' TpsCpoGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTps
TpsTpsTpsGpsTpsCpsGpsT 3';
wherein:
po represents a phosphodiester internucleotide linkage; and
ps represents a phosphorothioate internucleotide linkage.
10 . The ODN construct of claim 1 , having the structure of:
(1)
(SEQ ID NO: 27)
5′ Toco-po(HEG)po(HEG)po-TpsCpoGpsTpsCpoGpsTpsTps
TpsTpsGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTpsT 3′;
(2)
(SEQ ID NO: 28)
5′ Toco-po(HEG)po(HEG)po-TpsCpsGpsTpsCpoGpsTpsTps
TpsTpsGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpsGpsTpsT 3′;
(3)
(SEQ ID NO: 29)
5′ TpsCpoGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTps
TpsTpsTpsGpsTpsCpoGpsTpsT-(HEG)po(HEG)po-Toco 3′;
(4)
(SEQ ID NO: 30)
5′ TpsCpsGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTps
TpsTpsTpsGpsTpsCpsGpsTpsT-(HEG)po(HEG)po-Toco 3′;
(5)
(SEQ ID NO: 31)
5′ Chol-po(HEG)po(HEG)po-TpsCpoGpsTpsCpoGpsTpsTps
TpsTpsGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTpsT 3′;
(6)
(SEQ ID NO: 32)
5′ Chol-po(HEG)po(HEG)poTpsCpsGpsTpsCpsGpsTpsTps
TpsTpsGpsTpsCpsGpsTpsTpsTpsTpsGpsTpsCpsGpsTpsT 3′;
(7)
(SEQ ID NO: 33)
5′ Chol-po(HEG)po(HEG)po-TpsCpoGpsTpsCpsGpsTpsTps
TpsTpsGpsTpsCpsGpsTpsTpsTpsTpsGpsTpsCpsGpsTpsT 3′;
(8)
(SEQ ID NO: 34)
5′ Chol-po(HEG)po(HEG)po-TpsCpoGpsTpsCpoGpsTpsTps
TpsTpsGpsTpsCpsGpsTpsTpsTpsTpsGpsTpsCpsGpsTpsT 3′;
(9)
(SEQ ID NO: 35)
5′ Chol-po(HEG)po(HEG)po-TpsCpoGpsTpsCpoGpsTpsTps
TpsTpsGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpsGpsTpsT 3′;
(10)
(SEQ ID NO: 36)
5′ Chol-po(HEG)po(HEG)po-TpsCpoGpsTpsCpoGpsTpsTps
TpsTpsGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTpsT 3′;
(11)
(SEQ ID NO: 37)
5′ TpsCpsGpsTpsCpsGpsTpsTpsTpsTpsGpsTpsCpsGpsTps
TpsTpsTpsGpsTpsCpsGpsTpsTps-(HEG)po(HEG)po-
Chol 3′;
(12)
(SEQ ID NO: 38)
5′ TpsCpoGpsTpsCpsGpsTpsTpsTpsTpsGpsTpsCpsGpsTps
TpsTpsTpsGpsTpsCpsGpsTpsTps-(HEG)po(HEG)po-
Chol 3′;
(13)
(SEQ ID NO: 39)
5′ TpsCpoGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpsGpsTps
TpsTpsTpsGpsTpsCpsGpsTpsTps-(HEG)po(HEG)po-
Chol 3′;
(14)
(SEQ ID NO: 40)
5′ TpsCpoGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTps
TpsTpsTpsGpsTpsCpoGpsTpsTps-(HEG)po(HEG)po-
Chol 3′;
(15)
(SEQ ID NO: 41)
5′ TpsCpsGpsTpsCpsGpsTpsTpsTpsTpsGpsTpsCpsGpsTps
TpsTpsTpsGpsTpsCpsGpsTpsTps-(HEG)po(HEG)po-
Toco 3′;
(16)
(SEQ ID NO: 42)
5′ Toco-po(HEG)po(HEG)po-TpsCpoGpsTpsCpoGpsTpsTps
TpsTpsGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTpsT 3′;
(17)
(SEQ ID NO: 43)
5′ TpsCpoGpsTpsCpsGpsTpsTpsTpsTpsGpsTpsCpsGpsTps
TpsTpsTpsGpsTpsCpsGpsTpsTps-(HEG)po(HEG)po-
Toco 3′;
(18)
(SEQ ID NO: 44)
5′ TpsCpoGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpsGpsTps
TpsTpsTpsGpsTpsCpsGpsTpsTps-(HEG)po(HEG)po-
Toco 3′;
(19)
(SEQ ID NO: 45)
5′ TpsCpoGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTps
TpsTpsTpsGpsTpsCpsGpsTpsTps-(HEG)po(HEG)po-
Toco 3′;
(20)
(SEQ ID NO: 46)
5′ TpsCpoGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTps
TpsTpsTpsGpsTpsCpoGpsTpsTps-(HEG)po(HEG)po-
Toco 3′;
(21)
(SEQ ID NO: 47)
5′ Toco-po(HEG)po(HEG)po-TpsCpsGpsTpsCpsGpsTpsTps
TpsTpsGpsTpsCpsGpsTpsTpsTpsTpsGpsTpsCpsGpsTpsT 3′;
(22)
(SEQ ID NO: 48)
5′ Toco-po(HEG)po(HEG)po-TpsCpoGpsTpsCpsGpsTpsTps
TpsTpsGpsTpsCpsGpsTpsTpsTpsTpsGpsTpsCpsGpsTpsT 3′;
(23)
(SEQ ID NO: 49)
5′ Toco-po(HEG)po(HEG)po-TpsCpoGpsTpsCpoGpsTpsTps
TpsTpsGpsTpsCpsGpsTpsTpsTpsTpsGpsTpsCpsGpsTpsT 3′;
(24)
(SEQ ID NO: 50)
5′ Toco-po(HEG)po(HEG)po-TpsCpoGpsTpsCpoGpsTpsTps
TpsTpsGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpsGpsTpsT 3′;
(25)
(SEQ ID NO: 51)
5′-TpsCpoGpsTpsCGpsTpsTpsTpsTpsGpsTpsCGpsTpsTpsTps
TpsGpsTpsCpoGpsTpsTps-HEGps-Chol 3′;
(26)
(SEQ ID NO: 52)
5′-TpsCpoGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTps
TpsTpsTpsGpsTpsCpsGpsTpsTps(HEG)po(HEG)po-Chol 3′;
(27)
(SEQ ID NO: 53)
5′-GpsGpsGpsGpsTpsCpsApsApsCpoGpsTpsTpsGpsApsGps
GpsGpsGpsGpsGps-HEGps-Chol 3′;
(28)
(SEQ ID NO: 54)
5′-TpsCpsCpsApsTpsGpsApsCpsGpsTpsTpsCpsCpsTpsGps
ApsCpsGpsTpsTps-HEGps-Chol 3′;
(29)
(SEQ ID NO: 55)
5′-TpsCpsCpsApsTpsGpsApsCpsGpsTpsTpsCpsCpsTpsGps
ApsCpsGpsTpsTps-HEGps-Toco 3′;
(30)
(SEQ ID NO: 56)
5′ Palmitoyl-po(HEG)po(HEG)po-TpsCpoGpsTpsCpoGps
TpsTpsTpsTpsGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGps
TpsT 3′;
wherein:
Chol represents cholesterol;
Toco represents tocopherol;
Palmitoyl represents a palmitoyl group;
HEG represents ((CH 2 ) 2 O) 6 ;
po represents a phosphodiester linkage; and
ps represents a phosphorothioate linkage,
wherein the tocopherol, the cholesterol, or the palmitoyl group is covalently linked to the (HEG) directly via a po or ps linkage, or indirectly via a second linker that is linked to (HEG) directly via a po or ps linkage.
11 . (canceled)
12 . An ODN construct having a structure of:
or a pharmaceutically acceptable salt thereof, wherein the Oligonucleotides represents a CpG ODN comprising at least one CpG motif having a po internucleotide liknage.
13 . The ODN construct of claim 12 , wherein the CpG ODN contains SEQ ID NO:1.
14 . The ODN construct of claim 13 , wherein the CpG ODN consists of
5′TpsCpoGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTpsTpsTpsTpsGpsTpsCpoGpsTpsT-3′.
15 . The ODN construct of claim 12 , having the following structure:
16 . An ODN construct having a structure of:
17 . A pharmaceutical composition comprising the ODN construct of claim 1 and a pharmaceutically acceptable carrier.
18 . A method of preparing a pharmaceutical composition, comprising combining the ODN construct of claim 1 with a pharmaceutically acceptable carrier.
19 . A method of stimulating an immune response in a subject in need hereof, comprising administering to the subject the pharmaceutical composition of claim 17 .
20 . A method of treating a disease in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of claim 17 , wherein the disease is selected from the group consisting of Hepatitis B virus (HBV) and cancer.
21 . The ODN construct of claim 1 , wherein at least one CpG motif comprises two, three or four CpG motifs each having the po intemucleotide linkage.Join the waitlist — get patent alerts
Track US2021196831A1 — get alerts on status changes and closely related new filings.
We store only your email — no account needed. See our privacy policy.