Antimicrobial conjugates

a technology of conjugates and antimicrobials, applied in the field of antimicrobial conjugates, can solve the problems of inherently unstable nanoparticle suspensions, the ability of nanoparticles to kill either mammalian cells or microorganisms, etc., and achieve the effect of killing or inhibiting growth and demonstrating antimicrobial activity

Inactive Publication Date: 2008-02-28
UCL BUSINESS PLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0053] The efficacy of the mixtures as antimicrobials depends on many factors. The choice of nanoparticle type, choice of photosensitiser, nanoparticle size, concentration of nanoparticles and concentration of photosensitiser may all influence antimicrobial activity. Thus individual combinations may have particularly advantageous effects. For example and without limitations, the following combinations have been found particularly effective against Staphylococcus aureus: [0054] 2 nm diameter gold nanoparticles at a concentration of 4×1013 [0055] particles / ml with toluidine blue O at a concentration of 20 μM. [0056] 15 nm diameter gold nanoparticles at a concentration of 1×1014 to 1×1015 particles / ml with toluidine blue O at a concentration of 20 to 50 μM. [0057] 2 nm diameter gold nanoparticles at a concentration of4×1011 to 4×1013 particles / ml with methylene blue at a concentration of 20 μM. [0058] b 15 nm diameter gold nanoparticles at a concentration of 1×1013 to 1×1015 particles / ml with methylene blue at a concentration of 20 μM. [0059] 2 nm diameter gold nanoparticles at a concentration of 4×1011 particles / ml with tin chlorin e6 at a concentration of 20 μg / ml. [0060] 2 nm gold nanoparticles at a concentration of 4×1013 particles / ml with nile blue sulphate at a concentration of 20 to 50 μM.
[0054] 2 nm diameter gold nanoparticles at a concentration of 4×1013
[0055] particles / ml with toluidine blue O at a concentration of 20 μM.
[0056] 15 nm diameter gold nanoparticles at a concentration of 1×1014 to 1×1015 particles / ml with toluidine blue O at a concentration of 20 to 50 μM.
[0057] 2 nm diameter gold nanoparticles at a concentration of4×1011 to 4×1013 particles / ml with methylene blue at a concentration of 20 μM.
[0058] b 15 nm diameter gold nanoparticles at a concentration of 1×1013 to 1×1015 particles / ml with methylene blue at a concentration of 20 μM.

Problems solved by technology

Although the phthalocyanine / nanogold / tetraoctylammonium bromide was found to increase singlet oxygen generation, it was not demonstrated that these particles were able to kill either mammalian cells or microbes.
Nanoparticle suspensions are inherently unstable, and the nanoparticles tend to associate, or clump together.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0151] Gold nanoparticles (2.0 nm diameter; British Biocell International) in water (15×1013 particles per ml) were mixed with an equal volume of an aqueous solution of toluidine blue O (40 μM) and left at room temperature for 15 minutes. 100 μl of the gold-TB solution was added to 100 μl of a suspension of Staphylococcus aureus NCTC 6571 in phosphate buffered saline (PBS) and this was irradiated with white light from a fluorescent white lamp for 10 minutes. Controls consisted of:

[0152] (i) TB (final concentration=10 μM) and bacteria, irradiated for the same period of time,

[0153] (ii) nanogold (diluted 1:1 with water) and bacteria, irradiated for the same period of time,

[0154] (iii) bacteria without TB or nanogold, not irradiated (“control”).

[0155] After irradiation, the number of surviving bacteria was determined by viable counting. The results of the experiments (carried out twice with duplicate counts on each occasion) are shown in Table 1. The gold nanoparticles alone when i...

example 2

Production of Water-Soluble Gold Nanoparticles

[0156] HAuCl4.3H2O (42 mg, 0.11 mmol) was dissolved in deionised water (25 ml) to form solution A (˜5 mM). Na3C6H5O7.2H2O (125 mg, 0.43 mmol) was dissolved in deionised water (25 ml) to give solution B (˜20 mM). Solution A (1 ml) was stirred with deionised water (18 ml) and boiled for 2 min. Then solution B (1 ml) was added dropwise over a period of approximately 50 sec. causing the colour change from clear to blue to pink / purple. After a further 1 min. of heating, the solution was left to cool to room temperature. Two batches of nanogold particles were used for subsequent antibacterial assays—these are designated NN1 and NN2. The absorption spectrum of NN2 showed the wavelength of maximum absorption, λmax to be 527 nm. Batch NN1 had a λmax of 522 nm. Particle size analysis (position of UV plasmon absorption band measured using transmission electron microscope) of batch NN1 gave an average diameter of 14.76±2.34 nm.

Effect of Concentr...

example 3

[0159] The method of Example 2 was repeated using gold nanoparticles of 2 nm diameter (British Biocell International). The final concentration of nanogold used was 4×1013 particles / ml. TB was used at a final concentration of 10, 20 or 50 μM. The results are shown in Table 2 below. When the 2 nm nanogold particles were used, enhancement of lethal photosensitisation was evident using 20 μM TB but not when either 10 μM or 50 μM TB was used.

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Abstract

The present invention presents a metallic nanoparticle-ligand-photosensitiser conjugate, a method of making such conjugate, and a method of using such mixture for killing or preventing the growth of microbes.

Description

CLAIM OF BENEFIT OF FILING DATE [0001] This application claims the benefit of U.S. Provisional Patent Application Ser. No. 60 / 821,423 titled: “Antimicrobial Mixtures” filed on Aug. 4, 2006, U.S. Provisional Patent Application Ser. No. 60 / 868,130 titled: “Antimicrobial Conjugates” filed on Dec. 1, 2006, and United Kingdom Patent Application No. 0712287.2 titled: “Antimicrobial Conjugates” filed on Jun. 22, 2007.FIELD OF INVENTION [0002] The present invention relates to mixtures comprising charge-stabilized metallic nanoparticles and a photosensitiser, and their use as light activated antimicrobials. The present invention also relates to metallic nanoparticle-ligand-photosensitiser conjugates and their use as light activated antimicrobials. BACKGROUND OF THE INVENTION [0003] Photosensitisers, such as toluidine blue O, act as light-activated antimicrobial agents. Although they may have no antimicrobial activity at low concentrations in the dark, when irradiated with light of a certain ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K9/16A61P35/00A61Q11/00A61K33/242
CPCA01N59/16Y10S977/777A61K9/0031A61K9/0034A61K9/0043A61K9/0046A61K9/0053A61K9/10A61K31/28A61K41/0038A61K41/0057A61L15/00A61L26/00A61K9/0014C10M2219/108C01P2004/64A61K33/24A61K31/5415A61K9/14A01N2300/00A01N25/02A01N25/34A01N43/84A61K33/242A61K33/38A61K33/34A61K33/06A61P35/00A61K2300/00
Inventor WILSON, MIKEPARKIN, IVANNAIR, SEANGIL-TOMAS, JESUS
Owner UCL BUSINESS PLC
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