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US 2023/0383328 A1Published application
Methods and compositions for the enzymatic production of pseudouridine triphosphate
AssigneeNatures's Toolbox, Inc.
InventorsAlexander Koglin, Michael Humbert, Matthias Strieker, Grant Eliason
Life of this patent
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Abstract
The present invention includes novel systems, methods, and compositions for the enzymatic/chemical production of pseudouridine (Ψ) and its variants, such as N1-methyl-pseudouridine-5′-triphosphate (m1ΨTP).
Claims25
A system for the production of pseudouridine compounds comprising:
- a substrate comprising a quantity of isolated uracil nucleobase, and a quantity of isolated ribose-5-phosphate;
- a quantity of isolated pseudouridine-5′-phosphate glycosidase (PsuG) enzyme, or a fragment or variant thereof, wherein said PsuG catalyzes the formation of pseudouridine-5′-monophosphate (ΨMP) from the uracil nucleobase with a ribose-5-phosphate;
- a protecting agent that reacts with the hydroxyl groups of said ΨMP forming a protected ΨMP;
- a methylating agent that reacts with the protected ΨMP, forming a protected N1-methyl-pseudouridine-5′-monophosphate (protected m1ΨMP) compound;
- a deprotection agent that reacts with the protected m1ΨMP forming N1-methyl-pseudouridine-5′-monophosphate (m1ΨMP);
- and
- a phosphorylating agent that catalyzes the sequential phosphorylation of m1ΨMP forming N1-methyl-pseudouridine-5′-diphosphate (m1ΨDP), and N1-methyl-pseudouridine-5′-triphosphate (m1ΨTP).
4 more elements
20-22. (canceled)
24-107. (canceled)
A system for the production of pseudouridine compounds comprising:
- a substrate comprising a quantity of isolated uracil nucleobase, and a quantity of isolated ribose-5-phosphate;
- a quantity of isolated pseudouridine-5′-phosphate glycosidase (PsuG) enzyme, or a fragment or variant thereof, wherein said PsuG catalyzes the formation of pseudouridine-5′-monophosphate (ΨMP) from the uracil nucleobase with a ribose-5-phosphate;
- and
- a protecting agent that reacts with the hydroxyl groups of said ΨMP forming a protected ΨMP.
1 more element
A system for the production of pseudouridine compounds comprising:
- a substrate comprising a quantity of isolated uracil nucleobase, and a quantity of isolated ribose-5-phosphate;
- a quantity of isolated pseudouridine-5′-phosphate glycosidase (PsuG) enzyme, or a fragment or variant thereof, wherein said PsuG catalyzes the formation of pseudouridine-5′-monophosphate (ΨMP) from the uracil nucleobase with a ribose-5-phosphate;
- a protecting agent that reacts with the hydroxyl groups of said ΨMP forming a protected ΨMP;
- and
- a methylating agent that reacts with the protected ΨMP, forming a protected N1-methyl-pseudouridine-5′-monophosphate (protected m1ΨMP) compound.
2 more elements
A system for the production of pseudouridine compounds comprising:
- a substrate comprising a quantity of isolated uracil nucleobase, and a quantity of isolated ribose-5-phosphate;
- a quantity of isolated pseudouridine-5′-phosphate glycosidase (PsuG) enzyme, or a fragment or variant thereof, wherein said PsuG catalyzes the formation of pseudouridine-5′-monophosphate (ΨMP) from the uracil nucleobase with a ribose-5-phosphate;
- a protecting agent that reacts with the hydroxyl groups of said ΨMP forming a protected ΨMP;
- a methylating agent that reacts with the protected ΨMP, forming a protected N1-methyl-pseudouridine-5′-monophosphate (protected m1ΨMP) compound;
- and
- a deprotection agent that reacts with the protected m1ΨMP forming N1-methyl-pseudouridine-5′-monophosphate (m1ΨMP).
3 more elements
Dependencies are inferred from the claim text.