<title>Author Summary</title> <p>The human innate immune system, as the first line of defense against pathogens, is a vital component of our survival. One component of the innate immune system is the Toll-like receptor signaling network, which is responsible for transmitting activation signals from the outside of the cell to molecular machinery inside the cell. The innate immune system must be properly balanced, as excessive activation can lead to potentially lethal septic shock. Therefore, there is much interest in developing drugs that can mediate Toll-like receptor signaling so as to alleviate effects of excess activation. We present an <italic>in silico</italic> reconstruction of the Toll-like receptor signaling network and convert it into a mathematical framework that is suitable for constraint-based modeling and analysis. This approach leads to the identification of potential candidates for drug-based mediation. In addition to identifying targets for drug mediation of the Toll-like receptor network, we also supply a network model that may be continually updated and maintained.</p>