It has long been hypothesized that changes in gene regulation have played an important role in human evolution, but regulatory DNA has been much more difficult to study compared to protein-coding regions. Recent large-scale studies have created genome-scale catalogs of DNaseI Hypersensitive Sites (DHS), which demark potentially functional regulatory DNA. To better define regulatory DNA that has been subject to human specific adaptive evolution, we performed comprehensive evolutionary and population genetics analyses on over 18 million DHS discovered in 130 cell types. We identified 524 DHS that are conserved in non-human primates, but accelerated in the human lineage (haDHS), and estimate that 70% of substitutions in haDHS are attributable to positive selection. Through extensive computational and experimental analyses, we demonstrate that haDHS are often active in brain or neuronal cell types, play an important role in regulating the expression of developmentally important genes, including many transcription factors such as SOX6, POU3F2, and HOX genes, and identify striking examples of adaptive regulatory evolution that may have contributed to human specific phenotypes. More generally, our results reveal new insights into conserved and adaptive regulatory DNA in humans, and refine the set of genomic substrates that distinguish humans from their closest living primate relatives.