Methods to identify biologically active agents and synergistic combinations

Abstract

Methods are described using a particular type of assay system, the Multi-Pathway High Throughput Assay, in conjunction with a novel experimental strategy, whereby repeated cycles of experiments result in the identification of the most effective synergistic combinations of potential active agents from a library of materials. The novel experimental strategy not only requires far fewer total experiments than would be required using conventional discovery strategies but also maximizes the probability of finding highly synergistic combinations through the principle of multiple-pathway intervention.

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         42 . A method for identifying agents that display maximum biological activity in hair growth comprising testing the agents in the Low Temperature Whole Skin Organ Culture Assay said assay comprising a multiplicity of miniaturized cultures of skin maintained in a serum free growth medium at a temperature below about 30 C wherein said skin contains intact viable hair follicles below the sebaceous gland, and wherein hair growth is measured by a high-throughput hair growth measurement technique.  
     
     
         43 . The method according to  claim 42  wherein the high throughput hair growth measurement technique comprises the visualization of an incorporated fluorescent agent in the growing hair.  
     
     
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         47 . A method for identifying agents that display maximum biological activity in controlling acne comprising testing the agents in the Inhibition of IL1 Induced Hypercornification Assay said assay comprising either a viable culture of miniature pieces of epidermis with at least a portion of associated dermis in a multiple well assembly, or a culture of a sheet of viable epidermis and associated dermis, partitioned by the application of a surface barrier film into a pattern of isolated regions to which different test samples can be topically applied, in either case using a culture medium containing a level of IL1 sufficient to cause hypercornification; followed by the embedding of large numbers of cultured skin pieces in a single block and histological sectioning of the entire assembly and the automated histology, histochemistry or immunohistochemisty of histological sections carried out in a high throughput manner.  
     
     
         48 . A method for identifying agents that display maximum biological activity in controlling dental plaque comprising testing the agents in The Plaque Biofilm Regrowth Assay said assay comprising the growth of a plaque biofilm in multiwell plates under conditions of medium replenishment approximately simulating chemostat conditions, followed by the partial removal of the biofilm using mechanical abrasion in the presence of the test agents and detection of the rate of regrowth of the biofilm, optionally using a disclosing agent, via automated image detection.