High throughput screen for inhibitors of fatty acid biosynthesis in bacteria
AssigneeMillennium Pharmaceuticals, Inc., a Delaware corporation
InventorsChristopher Murphy, Philip Youngman
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Abstract
Methods for identifying compounds that are inhibitors of bacterial fatty acid biosynthesis are disclosed. Such compounds can be used as lead compounds in methods for preparing antibacterial agents for treating bacterial infections (e.g., in humans, animals, and plants). Inhibitors of bacterial fatty acid synthesis can also be tested for their ability to inhibit synthesis of acylated homoserine lactones. Compounds that inhibit synthesis of acylated homoserine lactones can be used as inhibitors of bacterial virulence. The disclosed methods allow for high throughput screening of libraries of test compounds.
Claims17
1-39. (canceled)
A composition comprising an antibacterial agent and a pharmaceutically acceptable carrier, wherein the antibacterial agent is identified by a method comprising:
- (i) contacting a bacterial cell with a test compound, wherein the bacterial cell contains
- (a) a promoter, the activity of which is increased in the presence of a compound that inhibits fatty acid biosynthesis (FAB), operably linked to
- (b) a reporter gene;
- (ii) measuring activity of the promoter, wherein an increase in activity, relative to the level of activity of the promoter in the absence of the test compound, indicates that the test compound is an inhibitor of bacterial FAB;
- and
- (iii) determining whether the inhibitor of FAB inhibits the growth of a bacterium, wherein a compound that inhibits the growth of a bacterium is an antibacterial agent.
A method of preparing an inhibitor of bacterial fatty acid biosynthesis (FAB), the method comprising:
- screening multiple test compounds by a method comprising:
- (i) contacting a bacterial cell with a test compound, wherein the bacterial cell contains
- (a) a promoter, the activity of which is increased in the presence of a compound that inhibits FAB, operably linked to
- (b) a reporter gene;
- and
- (ii) measuring activity of the promoter;
- and
- identifying candidate compounds that upregulate promoter activity;
- isolating one or more lead compounds from the candidate compounds;
- identifying and selecting a lead compound that inhibits bacterial FAB;
- and
- formulating the selected lead compound as an inhibitor of bacterial FAB.
A method of preparing an antibacterial agent, the method comprising:
- screening multiple test compounds by a method comprising:
- (i) contacting a bacterial cell with a test compound, wherein the bacterial cell contains
- (a) a promoter, the activity of which is increased in the presence of a compound that inhibits fatty acid biosynthesis (FAB), operably linked to
- (b) a reporter gene;
- (ii) measuring activity of the promoter;
- and
- identifying candidate compounds that upregulate promoter activity;
- isolating one or more lead compounds from the candidate compounds;
- identifying and selecting a lead compound that inhibits growth of a bacterium;
- and
- formulating the selected lead compound as an antibacterial agent.
A method for preparing an inhibitor of bacterial fatty acid biosynthesis (FAB), the method comprising:
- screening multiple test compounds by a method comprising:
- (i) contacting a bacterial cell with a test compound, wherein the bacterial cell contains
- (a) a promoter, the activity of which is increased in the presence of a compound that inhibits FAB, operably linked to
- (b) a reporter gene;
- and
- (ii) measuring activity of the promoter;
- and
- identifying candidate compounds that upregulate promoter activity;
- isolating one or more lead compounds from the candidate compounds;
- derivatizing said lead compound(s), thereby producing derivatives of said lead compound;
- identifying one or more derivatives that inhibit bacterial FAB;
- and
- formulating one or more derivatives as inhibitors of bacterial FAB.
A method for preparing an antibacterial agent, the method comprising:
- screening multiple test compounds by a method comprising:
- (i) contacting a bacterial cell with a test compound, wherein the bacterial cell contains
- (a) a promoter, the activity of which is increased in the presence of a compound that inhibits fatty acid biosynthesis (FAB), operably linked to
- (b) a reporter gene;
- (ii) measuring activity of the promoter;
- and
- identifying candidate compounds that upregulate promoter activity;
- isolating one or more lead compounds from the candidate compounds;
- derivatizing said lead compound(s), thereby producing derivatives of said lead compound;
- identifying one or more derivatives that inhibit growth of a bacterium;
- and
- formulating one or more derivatives as an antibacterial agent.
A composition comprising an inhibitor of bacterial virulence and a pharmaceutically acceptable carrier, wherein the inhibitor of bacterial virulence is identified by a method comprising:
- (i) contacting a bacterial cell with a test compound, wherein the bacterial cell contains
- (a) a promoter, the activity of which is increased in the presence of a compound that inhibits FAB, operably linked to
- (b) a reporter gene;
- (ii) measuring activity of the promoter, wherein an increase in activity, relative to the level of activity of the promoter in the absence of the test compound, indicates that the test compound is an inhibitor of bacterial FAB;
- and
- (iii) determining whether the inhibitor of FAB inhibits acylated homoserine lactone synthesis, wherein a compound that inhibits acylated homoserine lactone synthesis is an inhibitor of bacterial virulence.
A method of preparing an inhibitor of bacterial virulence, the method comprising:
- screening multiple test compounds by a method comprising:
- (i) contacting a bacterial cell with a test compound, wherein the bacterial cell contains
- (a) a promoter, the activity of which is increased in the presence of a compound that inhibits FAB, operably linked to
- (b) a reporter gene;
- (ii) measuring activity of the promoter, wherein an increase in activity, relative to the level of activity of the promoter in the absence of the test compound, indicates that the test compound is an inhibitor of bacterial FAB;
- and
- (iii) determining whether the inhibitor of FAB inhibits acylated homoserine lactone synthesis;
- and
- identifying candidate compounds that inhibit acylated homoserine lactone synthesis;
- isolating one or more lead compounds from the candidate compounds;
- identifying and selecting a lead compound that inhibits bacterial virulence;
- and
- formulating the selected lead compound as an inhibitor of bacterial virulence.
A method for preparing an inhibitor of bacterial virulence, the method comprising:
- screening multiple test compounds by a method comprising:
- (i) contacting a bacterial cell with a test compound, wherein the bacterial cell contains
- (a) a promoter, the activity of which is increased in the presence of a compound that inhibits FAB, operably linked to
- (b) a reporter gene;
- (ii) measuring activity of the promoter, wherein an increase in activity, relative to the level of activity of the promoter in the absence of the test compound, indicates that the test compound is an inhibitor of bacterial FAB;
- and
- (iii) determining whether the inhibitor of FAB inhibits acylated homoserine lactone synthesis;
- and
- identifying candidate compounds that inhibit acylated homoserine lactone synthesis;
- isolating one or more lead compounds from the candidate compounds;
- derivatizing said lead compound(s), thereby producing derivatives of said lead compound;
- identifying one or more derivatives that inhibit bacterial virulence;
- and
- formulating one or more derivatives as an inhibitor of bacterial virulence.
Dependencies are inferred from the claim text.