Lipase inhibiting composition

A composition for treating dandruff comprising a surfactant and a non-polymeric compound;

• wherein said compound, when used at a concentration of about 0.001% or less, inhibits the activity of any Malassezia globosa lipase by at least 50%, as indicated by glycerol production, using the following method:
• a) Add 270 μl of 25 mM MES, pH 5.5, 90 mM NaCl, 8 mM KCl, and 1 mM CaCl2 to each well of a deep well 96-well plate;
• b) Add 10 μl of olive oil, diluted three-fold into dimethyl sulfoxide, to each well;
• c) Add 15 μl of a candidate inhibitor compound to each well;
• d) Add 5 μl of Malassezia globosa lipase polypeptide to each well;
• e) Cover the 96-well plate;
• f) Vigorously shake the 96-well plate for one hour at room temperature;
• g) Following the one-hour period, spin the 96-well plate briefly in a centrifuge to force the liquid into the bottom of the well;
• h) After spinning, raise the pH in each well by adding 50 μl of 1 M Tris HCl pH 8.0;
• i) Transfer 90 μl of the solution in each well to a standard microtiter plate;
• j) Prepare a glycerol detection solution by combining 100 mM Tris HCl pH 7.6, 10 mM MgCl2, 2 mM 4-aminoantipyrine, 3 mM N-ethyl-N-(3-sulfopropyl)-m anisidine, 1 mM adenosine 5′-triphosphate, 20 units/ml peroxidase, 8 units/ml glycerol-3-phosphate oxidase, and 0.5 units/ml glycerol kinase;
• k) Add 90 μl of said glycerol detection solution to each well of the standard microtiter plate;
• l) Rotate the standard microtiter plate on a lab rotator for 15 minutes at room temperature;
• m) Measure the optical density at 540 nm, this is as an indication of the amount of glycerol produced;
• n) Generate a standard curve with known glycerol concentrations to know the absolute amount of glycerol generated;
• o) Compare the level of glycerol produced in step “m” to the standard curve.

A method to inhibit the activity of any Malassezia globosa lipase by at least 50% comprising applying a compound having a weight average molecular weight of less than about 1000 to the skin;

• wherein said compound, when used at a concentration of about 0.001% or less, inhibits the activity of any Malassezia globosa lipase by at least 50%, as indicated by glycerol production, using the following method:
• a) Add 270 μl of 25 mM MES, pH 5.5, 90 mM NaCl, 8 mM KCl, and 1 mM CaCl2 to each well of a deep well 96-well plate;
• b) Add 10 μl of olive oil, diluted three-fold into dimethyl sulfoxide, to each well;
• c) Add 15 μl of a candidate inhibitor compound to each well;
• d) Add 5 μl of Malassezia globosa lipase polypeptide to each well;
• e) Cover the 96-well plate;
• f) Vigorously shake the 96-well plate for one hour at room temperature;
• g) Following the one-hour period, spin the 96-well plate briefly in a centrifuge to force the liquid into the bottom of the well;
• h) After spinning, raise the-pH in each well by adding 50 μl of 1 M Tris HCl pH 8.0;
• i) Transfer 90 μl of the solution in each well to a standard microtiter plate;
• j) Prepare a glycerol detection solution by combining 100 mM Tris HCl pH 7.6, 10 mM MgCl2, 2 mM 4-aminoantipyrine, 3 mM N-ethyl-N-(3-sulfopropyl)-m anisidine, 1 mM adenosine 5′-triphosphate, 20 units/ml peroxidase, 8 units/ml glycerol-3-phosphate oxidase, and 0.5 units/ml glycerol kinase;
• k) Add 90 μl of said glycerol detection solution to each well of the standard microtiter plate;
• l) Rotate the standard microtiter plate on a lab rotator for 15 minutes at room temperature;
• m) Measure the optical density at 540 nm, this is as an indication of the amount of glycerol produced;
• n) Generate a standard curve with known glycerol concentrations to know the absolute amount of glycerol generated;
• o) Compare the level of glycerol produced in step “m” to the standard curve.

A method to inhibit the activity of any Malassezia globosa lipase by at least 50% comprising applying a non-polymeric compound to the skin;

• wherein said compound, when used at a concentration of about 0.001% or less, inhibits the activity of any Malassezia globosa lipase by at least 50%, as indicated by glycerol production, using the following method:
• a) Add 270 μl of 25 mM MES, pH 5.5, 90 mM NaCl, 8 mM KCl, and 1 mM CaCl2 to each well of a deep well 96-well plate;
• b) Add 10 μl of olive oil, diluted three-fold into dimethyl sulfoxide, to each well;
• c) Add 15 μl of a candidate inhibitor compound to each well;
• d) Add 5 μl of Malassezia globosa lipase polypeptide to each well;
• e) Cover the 96-well plate;
• f) Vigorously shake the 96-well plate for one hour at room temperature;
• g) Following the one-hour period, spin the 96-well plate briefly in a centrifuge to force the liquid into the bottom of the well;
• h) After spinning, raise the pH in each well by adding 50 μl of 1 M Tris HCl pH 8.0;
• i) Transfer 90 μl of the solution in each well to a standard microtiter plate;
• j) Prepare a glycerol detection solution by combining 100 mM Tris HCl pH 7.6, 10 mM MgCl2, 2 mM 4-aminoantipyrine, 3 mM N-ethyl-N-(3-sulfopropyl)-m anisidine, 1 mM adenosine 5′-triphosphate, 20 units/ml peroxidase, 8 units/ml glycerol-3-phosphate oxidase, and 0.5 units/ml glycerol kinase;
• k) Add 90 μl of said glycerol detection solution to each well of the standard microtiter plate;
• l) Rotate the standard microtiter plate on a lab rotator for 15 minutes at room temperature;
• m) Measure the optical density at 540 nm, this is as an indication of the amount of glycerol produced;
• n) Generate a standard curve with known glycerol concentrations to know the absolute amount of glycerol generated;
• o) Compare the level of glycerol produced in step “m” to the standard curve.