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US 2025/0108066 A1Published application
Trinucleotide cap analogs and methods of use thereof
AssigneeTriLink BioTechnologies, LLC
InventorsChunping XU, Michael HOUSTON, Alexandre LEBEDEV, Jordana Michelle HENDERSON, Andrew UJITA, Inna KOUKHAREVA, Chanfeng ZHAO, Ilya Vladimirovich ILICHEV
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Abstract
Described herein are novel trinucleotide cap analogs and methods of making and using the same. Also described herein is an RNA molecule comprising a 5′-cap, wherein the 5′-cap includes a trinucleotide cap analog as described herein. Methods of inducing a therapeutic effect in a 5 subject are also described herein, the methods including a step of administering to the subject an RNA molecule including the trinucleotide cap analog.
Claims70
A compound selected from the group consisting of:
- or a stereoisomer thereof, wherein:
- is a single bond or a double bond;
- R1 is H or CH3;
- R2 is H and R3 is OCH3 or F, or R2 and R3 are covalently bonded together and, together with intermediate atoms, form a 2′-O, 4′-C methylene bridge;
- X1 is O or CH;
- X2 is CH2 or CH;
- X3 is O or S;
- each independent Y is H+ or a cation;
- and
- n is 0, 1, 2, 3, or 4,
- wherein when X1 is CH, X2 is CH and is a double bond;
- or a stereoisomer thereof, wherein:
- R1 is H or CH3;
- X1 is O or S;
- each independent Y is H+ or a cation;
- and
- n is 0, 1, 2, 3, or 4,
- wherein when X1 is S, R1 is CH3;
- or a stereoisomer thereof, wherein:
- R1 is H or CH3;
- each independent Y is H+ or a cation;
- and
- n is 0, 1, 2, 3, or 4;
- or a stereoisomer thereof, wherein:
- R1 is H or CH3;
- X1, X2, and X3 are each independently selected from O and S, wherein when one of X1, X2, or X3 is S, the remaining of X1, X2, and X3 are O;
- each independent Y is H+ or a cation;
- and
- n is 0, 1, 2, 3, or 4;
- or a stereoisomer thereof, wherein:
- R1 is H or CH3;
- R2 is H and R3 is F, or R2 and R3 are covalently bonded together and, together with intermediate atoms, form a 2′-O, 4′-C methylene bridge;
- each independent Y is H+ or a cation;
- and
- n is 0, 1, 2, 3, or 4,
- wherein when R2 is H and R3 is F, R1 is not H;
- or a stereoisomer thereof, wherein:
- R1 is H or CH3;
- X1, X2, X3, and X4 are each independently selected from O and S;
- each independent Y is H+ or a cation;
- and
- n is 0, 1, 2, 3, or 4,
- wherein one of X1, X2, X3, and X4 is S;
- or a stereoisomer thereof, wherein:
- R1 and R2 are each independently selected from H and CH3;
- each independent Y is H+ or a cation;
- and
- n is 0, 1, 2, 3, or 4,
- wherein at least one of R1 or R2 is CH3;
- or a stereoisomer thereof, wherein:
- R1 and R2 are each independently selected from H and CH3;
- each independent Y is H+ or a cation;
- and
- n is 0, 1, 2, 3, or 4;
- or a stereoisomer thereof, wherein:
- R1 is H or CH3;
- R2 is OH, F, substituted or unsubstituted alkoxy, or thio;
- R3 is H or CH3;
- each independent Y is H+ or a cation;
- and n is 0, 1, 2, 3, or 4,
- wherein when R2 is OH, R1 is not H;
- or a stereoisomer thereof, wherein:
- R1 is H and CH3;
- each independent Y is H+ or a cation;
- and
- n is 0, 1, 2, 3, or 4;
- or a stereoisomer thereof, wherein:
- R1 is H and CH3;
- each independent Y is H+ or a cation;
- and
- n is 0, 1, 2, 3, or 4;
- or a stereoisomer thereof, wherein:
- R1 and R2 are each independently selected from H and CH3;
- each independent Y is H+ or a cation;
- and
- n is 0, 1, 2, 3, 4, or 5;
- or a stereoisomer thereof;
- and
- or a stereoisomer thereof.
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64-69. (canceled)
A method of administering to an animal a therapeutic dose unit of an mRNA molecule comprising a 5′-cap, wherein the 5′-cap comprises a compound selected from the group consisting of:
- or a stereoisomer thereof,
- wherein the therapeutic dose unit of the mRNA molecule comprises less than 7 ng/μg of double stranded RNA (dsRNA),
- wherein the subject exhibits increased tolerability to the administered therapeutic dose unit of the mRNA molecule as compared to an equivalent therapeutic dose unit of the mRNA molecule comprising 7 ng/μg or greater dsRNA.
71-74. (canceled)
A method for increasing in vivo translation of a polypeptide in a subject, comprising administering to the subject a therapeutic dose unit comprising an effective amount of an mRNA encoding a polypeptide, wherein the mRNA comprises a 5′-cap having the following formula:
- or a stereoisomer thereof,
- wherein the administered therapeutic dose unit is at least 20% lower than the therapeutic dose unit required to elicit the same response in the subject when administered a comparative mRNA encoding the polypeptide, wherein the comparative mRNA comprises a 5′-cap having the following formula:
- or a stereoisomer thereof;
- or
- a method for increasing in vivo translation of a polypeptide in a subject, comprising administering to the subject a therapeutic dose unit comprising an effective amount of an mRNA encoding a polypeptide, wherein the mRNA comprises a 5′-cap having the following formula:
- or a stereoisomer thereof,
- wherein the administered therapeutic dose unit is at least 20% lower than the therapeutic dose unit required to elicit the same response in the subject when administered a comparative mRNA encoding the polypeptide, wherein the comparative mRNA comprises a 5′-cap having the following formula:
- or a stereoisomer thereof;
- or
- a method for increasing in vivo translation of a polypeptide in a subject, comprising administering to the subject a therapeutic dose unit comprising an effective amount of an mRNA encoding a polypeptide, wherein the mRNA comprises a 5′-cap having the following formula:
- or a stereoisomer thereof,
- wherein the administered therapeutic dose is at least 20% lower than the therapeutic dose unit required to elicit the same response in the subject when administered a comparative mRNA encoding the polypeptide, wherein the comparative mRNA comprises a 5′-cap having the following formula:
- or a stereoisomer thereof;
- or
- a method for increasing in vivo translation of a polypeptide in a subject, comprising administering to the subject a therapeutic dose unit comprising an effective amount of an mRNA encoding a polypeptide, wherein the mRNA comprises a 5′-cap having the following formula:
- or a stereoisomer thereof,
- wherein the administered therapeutic dose unit is at least 20% lower than the therapeutic dose unit required to elicit the same response in the subject when administered a comparative mRNA encoding the polypeptide, wherein the comparative mRNA comprises a 5′-cap having the following formula:
- or a stereoisomer thereof.
15 more elements
76-81. (canceled)
A method of synthesizing a trinucleotide compound, comprising:
- (1) mixing an N7-methyl-guanosine-5′-diphosphate of the following structure:
- wherein
- X1 and X2 are each independently selected from the group consisting of O and S;
- X3 is O;
- R1 and R2 are each independently selected from H, OH, N3, F, substituted or unsubstituted alkoxy, and thio, wherein R1 and R2 optionally combine to form a heterocycle, with an activating reagent and an imidazole to form an activated intermediate;
- and
- (2) adding a salt reagent and a dinucleotide of the following structure:
- wherein
- is a single bond or a double bond;
- X4 and X6 are each independently selected from the group consisting of O and S;
- X5 is O, S, or CH;
- X7 is CH or CH2;
- R3 is OH, OMe, F and R4 is H, or wherein R3 and R4 are covalently bonded together and, together with intermediate atoms, form a 2′-O, 4′-C methylene bridge;
- and
- B1 and B2 are each independently selected from the group consisting of a purine ring and a pyrimidine ring
- to the activated intermediate to form the trinucleotide compound of the following structure:
- wherein the method is a one-pot synthesis;
- or
- a method of synthesizing a trinucleotide compound, comprising:
- (1) mixing a dinucleotide of the following structure:
- wherein
- is a single bond or a double bond;
- X4 is O;
- X6 is O or S;
- X5 is O, S, or CH;
- X7 is CH or CH2;
- R3 is OH, OMe, F and R4 is H, or wherein R3 and R4 are covalently bonded together and, together with intermediate atoms, form a 2′-O, 4′-C methylene bridge;
- and
- B1 and B2 are each independently selected from the group consisting of a purine ring and a pyrimidine ring,
- with an activating reagent and an imidazole to form an activated phosphate imidazolide of the following structure:
- and
- (2) adding a salt reagent and a compound of the following structure:
- wherein
- X1 and X2 are each independently selected from the group consisting of O and S;
- X3 is S;
- R1 and R2 are each independently selected from H, OH, N3, F, substituted or unsubstituted alkoxy, and thio, wherein R1 and R2 optionally combine to form a heterocycle, to the activated phosphate imidazolide to form the trinucleotide compound of the following structure:
- or
- a method of synthesizing a trinucleotide compound, comprising:
- (1) mixing a diphosphate dinucleotide of the following structure:
- wherein
- is a single bond or a double bond;
- X3 is O;
- X4 and X6 are each independently selected from O and S;
- X5 is O, S, or CH:
- X7 is CH or CH2;
- R3 is OH, OMe, F and R4 is H, or wherein R3 and R4 are covalently bonded together and, together with intermediate atoms, form a 2′-O, 4′-C methylene bridge;
- and
- B1 and B2 are each independently selected from the group consisting of a purine ring and a pyrimidine ring,
- with an activating reagent and an imidazole to form an activated phosphate imidazolide of the following structure:
- and
- (2) adding a salt reagent and a compound of the following structure:
- wherein
- X1 and X2 are each independently selected from O and S;
- and
- R1 and R2 are each independently selected from H, OH, N3, F, substituted or unsubstituted alkoxy, and thio, wherein R1 and R2 optionally combine to form a heterocycle, to the activated phosphate imidazolide to form the trinucleotide compound of the following structure:
- wherein the method is a one-pot synthesis.
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Dependencies are inferred from the claim text.