Antibacterial composition effective in treating gram negative bacterial infections and method for preparing the same

Inventors

Yoon, Seong Jun • JUN, Soo Youn • PARK, Jeong Won • Kim, Ji Hyun • KIM, Saet Byeol • Kang, Sang Hyeon

Assignees

Intron Biotechnology Inc

Abstract

A pharmaceutical composition for treating Gram negative bacteria-associated infections includes an antibacterial protein that includes at least one selected from the group of a protein having the amino acid sequence as set forth in SEQ ID NO: 1, a protein having the amino acid sequence as set forth in SEQ ID NO: 2, a protein having the amino acid sequence as set forth in SEQ ID NO: 3, a protein having the amino acid sequence as set forth in SEQ ID NO: 4, a protein having the amino acid sequence as set forth in SEQ ID NO: 5, a protein having the amino acid sequence as set forth in SEQ ID NO: 6, a protein having the amino acid sequence as set forth in SEQ ID NO: 7, and a protein having the amino acid sequence as set forth in SEQ ID NO: 8. A method of preparing the antibacterial protein is also disclosed.

Core Innovation

The document describes engineered antibacterial proteins (AP-1 to AP-8; SEQ ID NOs 1 to 8) that exhibit Gram-negative lytic activity. The engineered proteins are characterized as having enhanced outer-membrane penetration efficiency for Gram-negative bacteria including Acinetobacter baumannii, Pseudomonas aeruginosa, and Klebsiella pneumoniae, and are provided in pharmaceutical compositions for Gram-negative bacteria-associated infections.

The problem addressed is the treatment of Gram negative bacteria-associated infections using antibacterial proteins that can effectively lyse Gram-negative bacteria, including antibiotic-resistant strains. The document situates this problem within Gram-negative infections such as pneumonia, peritonitis, urinary tract infections, bloodstream infections, wound or surgical site infections, and meningitis. The disclosed engineered proteins are presented as achieving bacteriolysis against relevant Gram-negative targets.

The document also describes pharmaceutical compositions comprising an antibacterial protein defined by at least one of the amino acid sequences set forth in SEQ ID NOs 1 to 8. The compositions include formulation constraints such as protein concentration and pH, and may include excipients such as L-histidine and polysorbate/poloxamer and sorbitol or mannitol.

Experimental findings reported in the document include formulation stability assessment and antibacterial activity testing. The antibacterial activity is described as limited to the target Gram-negative species, with strong bacteriolysis against antibiotic-resistant strains including Meropenem-resistant Acinetobacter baumannii, and therapeutic efficacy in a mouse systemic infection model using AP-6, including improved survival and reduced observable responses versus control.

Claims Coverage

The provided dataset identifies two independent claims. The first independent claim covers a pharmaceutical composition for treating Gram negative bacteria-associated infections defined by antibacterial protein sequence membership (SEQ ID NOs 1 to 8). The second independent claim covers a method of preparing the antibacterial protein, linking specific antibacterial protein sequence membership (SEQ ID NOs 1 to 8) with culturing Escherichia coli cells carrying plasmid sequences (SEQ ID NOs 17 to 24) and subsequent inclusion-body recovery, solubilization, purifying, and refolding.

Overall, the claim coverage centers on a Gram-negative infection treatment pharmaceutical composition defined by antibacterial protein amino acid sequence membership (SEQ ID NOs 1 to 8) and an antibacterial protein preparation workflow in which sequence-defined proteins are produced in Escherichia coli via plasmids containing sequences (SEQ ID NOs 17 to 24), then processed through inclusion-body recovery, solubilization, purifying, and refolding.

Stated Advantages

Documented Applications