Can I use AutoRoute Express 2000 on Windows 10?
No, the Microsoft AutoRoute Express Europe 2000 won't work on Windows 10 because it doesn't have compatibility with the operating systems listed for the application. According with the description, you need Windows NT or Windows 95 and Windows 10 won't run apps developed for this operating systems.
You will have to either downgrade to previous version of Windows that you know it worked or virtualize Windows XP in a virtual machine with the help of VirtualBox.
Note: You will also need a Windows XP ISO or installation disc.