Microencapsulation of bacteriophages and related products
Inventors
Tawil, Nancy • ARNOLD, Edwige Caroline Rebecca • Katsarava, Ramaz • Tugushi, David • BERIDZE, Vakhtang
Assignees
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Publication Number
US-11413319-B2
Publication Date
2022-08-16
Expiration Date
Abstract
An encapsulated bacteriophage formulation and a method for encapsulating bacteriophages and bacteriophage-related products in polymeric microcapsules is provided. Some embodiments of the method of producing the encapsulated bacteriophages involves a water-in-oil-in-water double emulsion.
Core Innovation
The invention relates to a composition comprising polymer microcapsules and active bacteriophages encapsulated in the polymer microcapsules. The polymer microcapsules include an amino-acid based polymer, and the active bacteriophages are dispersed in the polymer microcapsules. The amino-acid based polymer is described as a poly(ester amide urea) comprising two blocks with random distribution, with an l:m block ratio constrained to range from 0.05:0.95 to 0.95:0.05, where l+m=1.
The document describes the amino-acid polymer structural definitions used for the two blocks, including that R1 is chosen from C2-C12 alkylenes optionally interrupted by oxygen, C3-C8 cycloalkylenes, or C3-C10 cycloalkylalkylenes. It further specifies that R3 is C2-C12 alkylene, and that R2 and R4 are independently selected from side chains of L- and D-amino acids so that the carbon to which R2 or R4 is attached has L or D chirality.
In the described embodiments, bacteriophage-loaded solid-state or suspension formulations are provided, including microcapsule suspensions stored for long periods with reported retention of encapsulated bacteriophages activity. The disclosure further includes an inorganic-salt approach in which bacteriophages adsorb onto CaCO3 or Ca3(PO4)2, with options described for encapsulation in amino-acid biodegradable polymer microparticles. Particle-size distributions after processing are reported, and bacteriophage activity is assessed using plaque assay to indicate preserved PFU activity after processing and storage.
Claims Coverage
The partial content identifies one independent claim directed to a bacteriophage-containing composition with amino-acid based polymer microcapsules, and dependent claims that refine polymer/capsule structure, physical size, and stability performance. The main inventive focus in the independent claim is the specific amino-acid based polymer architecture used in polymer microcapsules to encapsulate and disperse active bacteriophages, with defined block-ratio constraints and block structural selections.
Amino-acid based polymer microcapsules with dispersed active bacteriophages
A composition comprising polymer microcapsules, and active bacteriophages encapsulated in the polymer microcapsules, where the polymer microcapsules include an amino-acid based polymer and the active bacteriophages are dispersed in the polymer microcapsules.
Poly(ester amide urea) two-block architecture with random distribution
The amino-acid based polymer is a poly(ester amide urea) comprising the following two blocks with random distribution thereof.
Constrained l:m block ratio for the two blocks
wherein the ratio of l:m ranges from 0.05:0.95 to 0.95:0.05, l+m=1.
R1/R3/R2/R4 structural selections defining the amino-acid polymer blocks
R1 is chosen from C2-C12 alkylenes optionally interrupted by at least one oxygen, C3-C8 cycloalkylenes, C3-C10 cycloalkylalkylenes; R3 is C2-C12 alkylene; and R2 and R4 are independently chosen from the side chains of L- and D-amino acids so that the carbon to which R2 or R4 is attached has L or D chirality.
Overall, the claim coverage in the provided independent claim is concentrated on polymer microcapsules containing active bacteriophages dispersed within an amino-acid based poly(ester amide urea) having a two-block, random-distribution architecture. The structural constraints focus on the l:m block ratio and the defined selections for R1, R3, and the L/D-configured side-chain based groups R2 and R4, while dependent claim examples refine microcapsule size and stability metrics tied to stored bacteriophage activity.
Stated Advantages
Protection from solvent/acid/enzymatic inactivation.
Tunable release by microcapsule size and/or thickness.
Long-term phage activity and stability at 4°C and room temperature, up to one year.
Active bacteriophages remain active after storing the composition for one year at 4°C, with at least 1% remaining active.
Documented Applications
Bacteriophage-loaded solid-state or suspension formulations.
Bacteriophages adsorb onto CaCO3 or Ca3(PO4)2.
Encapsulation in amino-acid biodegradable polymer microparticles.
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