The first part of your regex looks incorrect:
(?!=\s{0,3}
should be (?!\s{0,3}
You are also looking ahead from the wrong place, so you should move this:
(?!\s{0,3}[Bb][Oo][Xx])
to just after you have found "PO" or "P.O." etc.
So your regex looks like this:
((([Cc]ustomer|[Cc]ust\.?) {0,5})?([Pp]\.? *[Oo]\.? *(?!\s{0,3}[Bb][Oo][Xx])|[Pp]urchase +[Oo]rder)) *([Nn]um\.?(ber)?|[Nn]o\.?)? *#? *:?
Also do your self a favor - unless you need case sensitivity, use case insensitivity, i.e. RegexOptions.IgnoreCase
and a more simple regex:
(((Customer|Cust\.?) {0,5})?(P\.? *O\.? *(?!\s{0,3}BOX)|Purchase +Order)) *(Num\.?(ber)?|No\.?)? *#? *:?