CpG oligonucleotide prodrugs, compositions thereof and associated therapeutic methods
Inventors
Verthelyi, Daniela • Beaucage, Serge L. • Grajkowski, Andrzej
Assignees
US Department of Health and Human Services
Publication Number
US-9809824-B2
Publication Date
2017-11-07
Expiration Date
2025-12-13
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Abstract
The present invention provides a CpG oligonucleotide prodrug that includes a thermolabile substituent on at least one nucleotide thereof. The present invention also provides compositions that include a carrier and a therapeutically effective amount of at least one CpG oligonucleotide prodrug. The present invention further provides therapeutic methods of using such thermolabile CpG oligonucleotide prodrugs and compositions thereof. The present invention further provides a method of inhibiting tetrad formation in a CpG oligonucleotide by functionalizing the CpG oligonucleotide with one or more thermolabile substituents.
Core Innovation
The invention provides CpG oligonucleotide prodrugs that are functionalized with thermolabile substituents bonded to at least one nucleotide, typically at the non-bridging oxygen atom of a phosphate or phosphorothioate group in the oligonucleotide. These thermolabile substituents are designed to cleave thermally in vivo, releasing the active parent CpG oligonucleotide to induce a desired immune response.
The prodrugs can be administered to a host as therapeutically effective immune stimulants that are rapidly internalized by immune cells and localized in endocytic vesicles where they interact with Toll-like receptor 9, triggering immunostimulatory cascades such as B-cell proliferation, dendritic cell maturation, and cytokine secretion. The thermolabile substituents can be structurally varied to control the onset and duration of action and to optimize physicochemical and pharmacological properties like cellular delivery and nuclease resistance.
The background describes a need for CpG oligonucleotides with improved delivery, resistance to nuclease cleavage, and controllable duration of action. Conventional CpG oligonucleotides are poorly taken up by cells, prone to tetrad formation that complicates manufacturing, and susceptible to degradation. The present invention addresses these problems by providing CpG oligonucleotides functionalized with thermolabile substituents that inhibit tetrad formation during manufacture and storage and act as prodrugs that release the active CpG oligonucleotide in vivo.
Claims Coverage
The patent includes one independent claim that defines the core composition and method of CpG oligonucleotide prodrugs with thermolabile substituents inhibiting tetrad formation and inducing cytokine production.
Thermolabile CpG oligonucleotide prodrug with poly-G tail functionalization inhibiting G-tetrad formation
A CpG oligonucleotide prodrug that has a poly-G tail at the 3′ end with thermolabile substituents bonded to the non-bridging oxygen atom of phosphate, phosphorothioate, or phosphoroselenoate diesters in the poly-G tail, in an amount sufficient to inhibit G-tetrad formation, where the thermolabile substituent is defined by a specific chemical formula.
Structural variations of the thermolabile substituent
The substituent can have variations including R2, R2′, R3 and R3′ as hydrogen; Z as CR4aR4a′ or CR4aR4a′CR4bR4b′ with all hydrogens; Z as NR4a with R4a as alkyl; Y as CH2R1 or C(O)H with R1 as H, OH, R1a, SR1a or NR1aR1a′, where R1a is alkyl, and R1a′ is hydrogen. Preferred methyl or tert-butyl groups are encompassed.
Inclusion of various CpG oligonucleotide types
The prodrug can include K-type, B-type, A-type, or D-type oligodeoxynucleotide sequences functionalized with the thermolabile substituents.
Pharmaceutical composition comprising the CpG oligonucleotide prodrug
Pharmaceutical compositions comprising a carrier and a therapeutically effective amount of the thermolabile CpG oligonucleotide prodrug are claimed.
Method of inducing cytokine production in a subject
A method comprising administering the pharmaceutical composition to induce production of cytokines such as interleukin-10, interferon-α, interferon-γ, or IL-6 in a subject.
The claims cover CpG oligonucleotide prodrugs functionalized with thermolabile substituents that inhibit G-tetrad formation, include specific structural details of substituents, encompass multiple CpG oligonucleotide types, pharmaceutical compositions thereof, and methods of inducing cytokine production.
Stated Advantages
Improved cellular delivery and nuclease resistance profile of CpG oligonucleotides.
Control over onset and duration of therapeutic efficacy through structural modifications of thermolabile substituents.
Avoidance of tetrad formation during manufacturing and storage, facilitating large-scale production.
Enhanced bioavailability and stability of the CpG oligonucleotide prodrugs.
Ability to delay or sustain the immune response as needed for specific therapeutic applications.
Documented Applications
Inducing immunostimulatory or immunomodulatory immune responses in a host.
Treatment or prevention of infections caused by parasites, bacteria, viruses, and bio-warfare agents.
Treatment or prevention of neoplasms and as an adjunct to chemotherapy or radiation.
Use as a vaccine adjuvant to improve vaccine efficacy against infectious diseases such as Leishmania, Hepatitis, HIV, and malaria.
Treatment or prevention of allergic reactions and allergic asthma in combination with anti-allergenic agents.
Ex vivo activation of immune cells such as monocytes, dendritic cells, lymphocytes, or natural killer cells for therapeutic administration.
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