Use & \phantom{{}= - \bigl[}
. Also, you should use &=
instead of =&
. Further, use \bigl[
and \bigr]
instead of \left[
and right]
. This doesn't need to be balanced like \left.
. Also the \\
in the last line shouldn't be used. In the last line, use {}+{}
so that the spacing around +
is correct.
\documentclass[]{report}
\usepackage{amsmath}
\begin{document}
\begin{align}
\dot{V} &= \frac{1}{\gamma_{2,1}}\sigma_{2,1} \dot{σ}_{2,1} + \frac{1}{\gamma_{2,2}}\sigma_{2,2} \dot{σ}_{2,2} + \frac{1}{\gamma_{2,3}}\sigma_{2,3} \dot{σ}_{2,3} \notag \\
&= {}-{} \bigl[\sigma_{2,1}^2 \sin^2(x_1) + \sigma_{2,2}^2 x_2^2 + \sigma_{2,3}^2 u^2 + 2\sigma_{2,1}\sigma_{2,2}\sin(x_1)x_2 + 2 \sigma_{2,1}\sigma_{2,3}\sin(x_1)u \notag \\
& \phantom{{}= - \bigl[} {}+{} 2 \sigma_{2,2}\sigma_{2,3}x_2 u \vphantom{\sigma_{2,1}^2} \bigr] \notag %% no \\ here
\end{align}
\end{document}

Here is another version with shorter second line as recommended by egreg.
\documentclass[]{report}
\usepackage{amsmath}
\begin{document}
\begin{align}
\dot{V} &= \frac{1}{\gamma_{2,1}}\sigma_{2,1} \dot{σ}_{2,1} + \frac{1}{\gamma_{2,2}}\sigma_{2,2} \dot{σ}_{2,2} + \frac{1}{\gamma_{2,3}}\sigma_{2,3} \dot{σ}_{2,3} \notag \\
&= {}-{} \bigl[\sigma_{2,1}^2 \sin^2(x_1) + \sigma_{2,2}^2 x_2^2 + \sigma_{2,3}^2 u^2 + 2\sigma_{2,1}\sigma_{2,2}\sin(x_1)x_2 \notag \\
& \phantom{{}= - \bigl[} {}+{} 2 \sigma_{2,1}\sigma_{2,3}\sin(x_1)u + 2 \sigma_{2,2}\sigma_{2,3}x_2 u \vphantom{\sigma_{2,1}^2} \bigr] \notag %% no \\ here
\end{align}
\end{document}

\quad
or\qquad
after the final&