diff --git a/Library/Homebrew/language/python_virtualenv_constants.rb b/Library/Homebrew/language/python_virtualenv_constants.rb
index a6bf86b2a0..e9a3103ea2 100644
--- a/Library/Homebrew/language/python_virtualenv_constants.rb
+++ b/Library/Homebrew/language/python_virtualenv_constants.rb
@@ -1,6 +1,6 @@
 PYTHON_VIRTUALENV_URL =
-  "https://files.pythonhosted.org/packages/33/bc" \
-  "/fa0b5347139cd9564f0d44ebd2b147ac97c36b2403943dbee8a25fd74012" \
-  "/virtualenv-16.0.0.tar.gz".freeze
+  "https://files.pythonhosted.org/packages/4e/8b" \
+  "/75469c270ac544265f0020aa7c4ea925c5284b23e445cf3aa8b99f662690" \
+  "/virtualenv-16.1.0.tar.gz".freeze
 PYTHON_VIRTUALENV_SHA256 =
-  "ca07b4c0b54e14a91af9f34d0919790b016923d157afda5efdde55c96718f752".freeze
+  "f899fafcd92e1150f40c8215328be38ff24b519cd95357fa6e78e006c7638208".freeze