Not natively, but ntvdmx64[0][1] and otvdm[2]/winevdm[3] work fine, albeit a bit slowly.

[0]: http://www.columbia.edu/~em36/ntvdmx64.html

[1]: https://github.com/leecher1337/ntvdmx64

[2]: http://www.columbia.edu/~em36/otvdm.html

[3]: https://github.com/otya128/winevdm

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For those interested, Wikipedia says there's no native/builtin 64bit NTVDM due to

> In an x86-64 CPU, virtual 8086 mode is available as a sub-mode only in its legacy mode (for running 16- and 32-bit operating systems), not in the native 64-bit long mode.

...

> The NTVDM is not supported on x86-64 editions of Windows,[31] including DOS programs,[32] because NTVDM uses VM86 CPU mode instead of the Local Descriptor Table in order to enable 16‑bits segment required for addressing[33] and AArch64 because Microsoft did not release a full emulator for this incompatible instruction set like it did on previous incompatible architecture.

https://en.wikipedia.org/wiki/Virtual_DOS_machine#Windows_NT...

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Personally, since Microsoft still releases 32bit builds of Windows 10, I just dual boot 32bit (~48GB)/64bit (~900GB) partitions and run the 16bit programs natively on an old Thinkpad.

32/64 bit is a compile time option not a separate codebase and unlike macOS they haven't discontinued 32 bit even in the latest versions.