Methods and compositions for the enzymatic production of pseudouridine triphosphate
AssigneeNatures's Toolbox, Inc.
InventorsAlexander Koglin, Michael Humbert, Matthias Strieker, Grant Eliason
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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).
Claims20
A method of synthesizing a pseudouridine compound comprising the step of protecting a pseudouridine-5′-monophosphate (ΨMP) with a protecting agent that reacts with the hydroxyl groups of said ΨMP forming a protected ΨMP.
A method of synthesizing a pseudouridine compound comprising:
- protecting a pseudouridine-5′-monophosphate (ΨMP) with a protecting agent that reacts with the hydroxyl groups of said ΨMP forming the protected ΨMP;
- and
- methylating the protected ΨMP with a methylating agent, forming a protected N1-methyl-pseudouridine-5′-monophosphate (protected m1ΨMP).
A method of synthesizing a pseudouridine compound comprising:
- methylating a protected ΨMP with a methylating agent forming a protected N1-methyl-pseudouridine-5′-monophosphate (protected m1ΨMP);
- and
- deprotecting the protected m1ΨMP with a deprotection agent forming a N1-methyl-pseudouridine-5′-monophosphate (m1ΨMP).
A method of synthesizing a pseudouridine compound comprising:
- deprotecting a protected m1ΨMP with a deprotection agent forming a N1-methyl-pseudouridine-5′-monophosphate (m1ΨMP);
- and
- phosphorylating the m1ΨMP with a phosphorylating agent forming N1-methyl-pseudouridine-5′-diphosphate (m1ΨDP) and N1-methyl-pseudouridine-5′-triphosphate (m1ΨTP).
A method of synthesizing a pseudouridine compound comprising:
- combining a uracil nucleobase and a ribose-5-phosphate to form pseudouridine-5′-monophosphate (ΨMP), wherein the formation of ΨMP is catalyzed by pseudouridine-5′-phosphate glycosidase, or a functional fragment or variant thereof;
- and
- protecting the ΨMP with a protecting agent that reacts with the hydroxyl groups of said ΨMP forming the protected ΨMP.
Dependencies are inferred from the claim text.