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Product and process for inhibition of biofilm development

a biofilm and process technology, applied in the direction of biocide, peptide/protein ingredients, catheter, etc., can solve the problems of undesirable biofilm development, inability to correct cf by gene therapy, etc., to and inhibit the formation or polymerization

Inactive Publication Date: 2006-02-09
NAT JEWISH HEALTH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0016] Another embodiment of the present invention relates to a composition for inhibiting biofilm formation or reducing biofilms in a subject. The composition can comprise any one, two, three, four, five, or more compounds as described above. In one embodiment, the composition includes: (1) a first compound that inhibits the formation or polymerization of actin microfilaments or depolymerizes actin microfilaments at or proximal to the site of biofilm formation; and (2) a second compound that is an anti-DNA compound. This composition can further include a compound that inhibits the accumulation of, necrosis of, or release of the cellular contents of, neutrophils at or proximal to the site of biofilm formation, and a carrier suitable for application to the site of biofilm formation. In another aspect, the composition can include: (1) a first compound that inhibits the formation or polymerization of actin microfilaments or depolymerizes actin microfilaments at or proximal to the site of biofilm formation; and (2) a second compound that inhibits the accumulation of, necrosis of, or release of the cellular contents of, neutrophils at or proximal to the site of biofilm formation, and a carrier suitable for application to the site of biofilm formation.
[0017] Yet another embodiment of the present invention relates to a method to iden

Problems solved by technology

Despite some promising advances, correction of CF by gene therapy is not yet attainable.
In addition to CF, a variety of other medical conditions and treatments can cause the undesirable development of biofilms.

Method used

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  • Product and process for inhibition of biofilm development
  • Product and process for inhibition of biofilm development
  • Product and process for inhibition of biofilm development

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0108] The following example demonstrates the effect of human neutrophils on initial P. aeruginosa biofilm development.

[0109] Referring to FIG. 1A, biofilm development of P. aeruginosa (□) was compared with P. aeruginosa in the presence of neutrophils (●). Neutrophil cytotoxicity (▴) equaled 92% after 24 hrs of exposure to P. aeruginosa (FIG. 1A; lot (right axis) depicts mean percent of viable neutrophils±SD (n=4)). In the presence of neutrophils, a reduction in the number of surviving P. aeruginosa was detected at 4 hours, while at later timepoints, the bactericidal effects of the neutrophil no longer reached significance (FIG. 1A; plot depicts mean±SD of CFU (n=4). *p<0.05 by Student's t-test).

[0110] When measured simultaneously, the number of viable P. aeruginosa in the planktonic state was significantly decreased by the presence of neutrophils, while the number of viable P. aeruginosa in the biofilm state was significantly increased (FIGS. 1B-C). Referring to FIG. 1B, the pres...

example 2

[0116] The following example shows the enhancement of P. aeruginosa biofilm formation by lysed neutrophils.

[0117] Since neutrophil necrosis is rapid in the presence of P. aeruginosa, the capacity of the cellular content of lysed neutrophils to evoke enhanced P. aeruginosa biofilm development was tested. Parameters of biofilm development of P. aeruginosa (□) were compared with P. aeruginosa in the presence of lysed neutrophils (●) at time intervals from 0 to 72 hrs. Combining P. aeruginosa with neutrophil lysates significantly enhanced biofilm formation as measured by CV staining and exopolysaccharide synthesis, and supplementing the early biofilm with additional quantities of lysed neutrophils at 24 and 48 hrs further enhanced biofilm production (FIGS. 2A,C). Referring to FIG. 2A, crystal violet staining of biofilm density demonstrated that the presence of lysed neutrophils resulted in a greater quantity of biofilm compared to P. aeruginosa in the absence of neutrophils by 4 hrs. W...

example 3

[0119] The following example demonstrates the enhancement of P. aeruginosa biofilm formation by isolated neutrophil components.

[0120] The data described in the Examples above indicates that neutrophil cellular contents are largely responsible for enhanced P. aeruginosa biofilm formation. Analysis of CF sputum demonstrates high concentrations of granule proteins, actin and DNA released from necrotic neutrophils (18, 28, 38). The capacity of each of these compounds to mediate enhanced early P. aeruginosa biofilm formation was tested. Referring to FIG. 3A, P. aeruginosa (▾) was combined with granule proteins (quantity equivalent to 5×106 neutrophils) compared to P. aeruginosa (□) in the absence of granule proteins and P. aeruginosa combined with live neutrophils (●). Supplementing planktonic P. aeruginosa with purified granule proteins failed to enhance biofilm production over a range of concentrations (FIG. 3A; Plot depicts mean±SD of O.D. crystal violet measurements (n=4). *p<0.05 b...

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Abstract

Disclosed are compositions and methods for the inhibition of biofilm formation or reduction of existing or developing biofilms in a patient. These methods also inhibit the aggregation of bacteria that form biofilms in the airways. The methods include administering to a subject that has or is at risk of developing biofilms a compound or formulation that inhibits the formation or polymerization of actin microfilaments or depolymerizes actin microfilaments at or proximal to the site of biofilm formation. Such a compound can be administered in combination with a compound or formulation that inhibits the accumulation or activity of cells that are likely to undergo necrosis at or proximal to the site of biofilm formation (i.e., neutrophils). The methods and compositions can further include the use of anti-DNA and / or anti-mucin compounds, as well as other therapeutic compounds and compositions.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of priority under 35 U.S.C. § 119(e) from U.S. Provisional Application Ser. No. 60 / 599,495, filed Aug. 6, 2004. The entire disclosure of U.S. Provisional Application Ser. No. 60 / 599,495 is incorporated herein by reference in its entirety.Government Support [0002] This invention was supported, in part, using funds provided by NIH Grant Nos. HL061407 and HL068743, each awarded by the National Institutes of Health, and using funds provided by NIAID Grant No. A115950, awarded by the National Institute of Allergy and Infectious Diseases. The government has certain rights to this invention.FIELD OF THE INVENTION [0003] The present invention generally relates to methods and compositions for the inhibition of biofilm formation or reduction of existing or developing biofilms in a patient. BACKGROUND OF THE INVENTION [0004] Cystic fibrosis (CF) lung disease features persistent neutrophil accumulation to the air...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K31/7076A61K31/7072A61K31/522
CPCA61K31/522A61K31/7072A61K31/7076A61K38/00A61K45/06A61L15/44A61L2300/432A61K9/007A61K39/395A61L29/16
Inventor NICK, JERRY A.WALKER, TRAVIS S.WORTHEN, G. SCOTT
Owner NAT JEWISH HEALTH
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