Gram-negative lysin-antimicrobial peptide (AMP) polypeptide constructs, lysins, isolated polynucleotides encoding same and uses thereof in human serum
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Publication Number
US-11773140-B2
Publication Date
2023-10-03
Expiration Date
Abstract
The present disclosure is directed to lysin-AMP polypeptide constructs, isolated lysin polypeptides, and pharmaceutical compositions comprising the isolated polypeptides and/or lysin-AMP polypeptide constructs. Methods of using the lysin-AMP polypeptide constructs, isolated lysin polypeptides and pharmaceutical compositions are also herein provided. In addition, isolated polynucleotides encoding the lysin-AMP polypeptide constructs and isolated lysin polypeptides are disclosed herein.
Core Innovation
The invention relates to lysin–AMP polypeptide constructs and isolated polynucleotides comprising nucleic acid molecules encoding them. The first component comprises a lysin with lytic activity and an α-helix domain at the C-terminus or the N-terminus, including selected lysin variants and active fragments retaining the α-helix domain, and the second component comprises at least one antimicrobial peptide (AMP), including specified AMP sequences or AMP activity polypeptides having at least 80% sequence identity.
The constructs are defined by sequence-identity relationships for retained lysin activity and AMP activity, with sequence identity ranges from at least 80% up to at least 99%. The disclosure further includes optional sequence-level constraints including pI-increasing mutation or point mutation options for selected lysins, and the α-helix domain placement is limited to the C-terminus or N-terminus of the lysin.
The document further describes pharmaceutical compositions comprising the lysin–AMP polypeptide construct and a pharmaceutically acceptable carrier, as well as recombinant vectors and host cells for expressing the encoded constructs. It also provides methods for inhibiting growth, reducing population, or killing at least one Gram-negative bacteria, including P. aeruginosa, in the presence or absence of human serum.
Claims Coverage
The partial content includes multiple independent claim sets directed to lysin–AMP polypeptide constructs, isolated polynucleotides encoding the constructs, pharmaceutical compositions, and methods for inhibiting Gram-negative bacteria. Across these claims, the inventive features center on combining a lysin retaining an α-helix domain with a specified AMP component, and on comparative effectiveness against P. aeruginosa, including in the presence or absence of human serum.
Lysin–AMP polypeptide construct with α-helix-retaining lysin and specified AMP component
A lysin–AMP polypeptide construct comprising a first component selected from lysins with an α-helix domain at the C-terminus or N-terminus, or a lytic polypeptide having at least 80% sequence identity retaining the α-helix domain, or an active fragment retaining the α-helix domain; and a second component comprising at least one antimicrobial peptide selected from a specified group or a polypeptide having AMP activity at least 80% identical to at least one specified AMP sequence.
Isolated polynucleotide encoding the lysin–AMP polypeptide construct
An isolated polynucleotide comprising a nucleic acid molecule encoding a lysin–AMP polypeptide construct, including a first nucleic acid molecule encoding a first component that is a selected lysin with an α-helix domain at the C-terminus or N-terminus, or a lytic activity polypeptide at least 80% identical retaining the α-helix domain, or an active fragment retaining the α-helix domain; and a second nucleic acid molecule encoding a second component comprising at least one specified AMP or an AMP-activity polypeptide at least 80% identical to at least one specified AMP sequence.
Pharmaceutical composition with pharmaceutically acceptable carrier
A pharmaceutical composition comprising a lysin–AMP polypeptide construct and a pharmaceutically acceptable carrier, where the construct includes the same defined first component and the same defined second component.
Comparative effectiveness of combination versus either antibiotic or lysin–AMP construct alone in human serum
A method of inhibiting the growth, reducing the population, or killing at least one Gram-negative bacteria where the Gram-negative bacteria is P. aeruginosa, comprising contacting the bacteria with a composition containing an effective amount of the lysin–AMP polypeptide construct, and requiring that administration of the combination is more effective than administration of either the antibiotic or the lysin–AMP polypeptide construct individually, in the presence or absence of human serum or both.
Across the independent claims, the document defines lysin–AMP polypeptide constructs by combining a lytic lysin with a specified AMP, claims isolated polynucleotides encoding these constructs, claims pharmaceutical compositions containing them with a pharmaceutically acceptable carrier, and claims methods demonstrating increased effectiveness against P. aeruginosa, including an explicit comparison in the presence or absence of human serum.
Stated Advantages
Administration of the combination is more effective in inhibiting growth, reducing population, or killing Gram-negative bacteria than administration of either the antibiotic or the lysin–AMP polypeptide construct individually.
More effective inhibition, reduction, or killing can be observed in the presence or absence of human serum.
Documented Applications
Treating a Gram-negative bacterial infection by administering a pharmaceutical composition comprising a lysin–AMP polypeptide construct to a subject diagnosed with, at risk for, or exhibiting symptoms of the infection.
Preventing or treating a bacterial infection by co-administering a first pharmaceutical composition containing the lysin–AMP polypeptide construct and a second effective amount of an antibiotic suitable for Gram negative bacterial infections.
Inhibiting the growth, reducing the population, or killing at least one Gram-negative bacteria where the Gram-negative bacteria is P. aeruginosa by contacting the bacteria with a composition containing an effective amount of the lysin–AMP polypeptide construct.
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