# US 10,499,655 B2 · Reagents and methods for inhibiting or disrupting biofilm

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- Number: US 10,499,655 B2
- Document ID: US10499655B2
- Status: Granted
- Kind: B2
- Application number: 15/557,812
- Filed: 2016-03-11
- Published: 2018-02-15
- Granted: 2019-12-10
- Earliest claimed: 2015-03-13
- Assignees: Institute of Microbiology, Chinese Academy of Sciences
- Applicants: Institute of Microbiology, Chinese Academy of Sciences
- Inventors: Lvyan Ma; Shan Yu; Huijun Wu; Shiwei Wang; Di Wang
- Web page: https://patentgrep.com/patent/US10499655B2

## CPC

- A01N 37/46: N-acyl derivatives
- A01N 43/60: 1,4-Diazines; Hydrogenated 1,4-diazines
- A01N 63/50: Isolated enzymes; Isolated proteins (peptides A01N37/46)
- A61K 38/164: {from bacteria}
- A61P 31/04: Antibacterial agents
- A01N: PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF (preservation of food or foodstuff A23); BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES (preparations for medical, dental or toiletry purposes which kill or prevent the growth or proliferation of unwanted organisms A61K); PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

The present disclosure provides an inhibitor for inhibiting or disrupting biofilm and use thereof. Specifically, the present disclosure an application of a PsIG protein and a coding sequence thereof in inhibiting or disrupting a biofilm of a microorganism. The present disclosure also provides a composition and a preparation containing the PsIG protein to inhibit or disrupt a biofilm of a microorganism. The experiment shows that PsIG can effectively inhibit or disrupt a biofilm of Pseudomonas species, suggesting the great potential of PsIG in clinical and environmental fields.

## Claims

### Claim 1

A method of inhibiting formation of biofilm on a surface, comprising contacting the surface with a composition comprising a PsIG protein and at least one antimicrobial agent, wherein the PsIG protein increases the activity of the antimicrobial agent to inhibit formation of the biofilm and consists of an amino acid sequence of SEQ ID NO: 2, wherein a concentration of the PsIG protein is in a range of 5 nM to 100 nM and has no cytotoxicity.

### Claim 2 (depends on 1)

The method of claim 1, wherein the at least one antimicrobial agent is selected from the group consisting of azithromycin, fleroxacin, ciprofloxacin, and tobramycin.

### Claim 3 (depends on 1)

The method of claim 1, wherein the at least one antimicrobial agent comprises fleroxacin.

### Claim 4 (depends on 1)

The method of claim 1, wherein the composition further comprises a pharmaceutically/environmentally acceptable carrier.

### Claim 5 (depends on 1)

The method of claim 1, wherein the composition is lyophilized.

### Claim 6 (depends on 1)

The method of claim 1, wherein the PsIG protein has an in vitro activity of degrading 75% Psi polysaccharide at 30° C. in one hour.

### Claim 7 (depends on 1)

The method of claim 1, wherein the biofilm is formed by Pseudomonas species or is contained Pseudomonas species.

### Claim 8 (depends on 1)

The method of claim 1, wherein the biofilm is formed on a surface of a medical device or a surface of a pipe.

### Claim 9 (depends on 1)

The method of claim 1, wherein the PsIG protein is at a concentration that has no cytotoxicity to Caco2, HT-29 cells, or nematodes.

### Claim 10

A method of disrupting a biofilm, comprising contacting the biofilm with a composition comprising a PsIG protein, wherein the PsIG protein consists of an amino acid sequence of SEQ ID NO: 2, wherein a concentration of the PsIG is in a range of 5 nM to 100 nM and has no cytotoxicity.

### Claim 11 (depends on 10)

The method of claim 10, further comprising contacting the biofilm with at least one antimicrobial agent.

### Claim 12 (depends on 11)

The method of claim 11, wherein the at least one antimicrobial agent is selected from the group consisting of azithromycin, fleroxacin, ciprofloxacin, and tobramycin.

### Claim 13 (depends on 11)

The method of claim 11, wherein the at least one antimicrobial agent comprises tobramycin.

### Claim 14 (depends on 10)

The method of claim 10, wherein the composition further comprises at least one of a pharmaceutically acceptable carrier and an environmentally acceptable carrier.

### Claim 15 (depends on 10)

The method of claim 10, wherein the composition is lyophilized.

### Claim 16 (depends on 10)

The method of claim 10, wherein the PsIG protein has an in vitro activity of degrading 75% Psi polysaccharide at 30° C. in one hour.

### Claim 17 (depends on 10)

The method of claim 10, wherein the biofilm is formed by Pseudomonas species or is contained Pseudomonas species.

### Claim 18 (depends on 10)

The method of claim 10, wherein the biofilm is formed on a surface of a medical device or a surface of a pipe.

### Claim 19 (depends on 10)

The method of claim 10, wherein the PsIG protein is at a concentration that has no cytotoxicity to Caco2, HT-29 cells, or nematodes.