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The <i>Streptomyces</i> genus has long been recognized as a prolific and valuable source of diverse secondary metabolites. These metabolites contribute significantly to the intricate chemical diversity exhibited by <i>Streptomyces</i>, making them an indispensable reservoir for drug discovery, agricultural applications, and industrial processes. Exploiting the potential of these natural compounds holds the promise of ushering in a new era in insect pest management, reducing reliance on synthetic chemicals and fostering ecologically sustainable solutions. This study dives into the realm of chemo diversity within isolates of <i>Streptomyces nojiriensis</i> and <i>Streptomyces novaecaesareae</i>, with a specific focus on the production of insecticidal compounds. We explored chromatographic techniques for the identification and isolation of insecticidal compounds, and two bioactive compounds were identified in extracts of <i>S. novaecaesareae</i>. Valinomycin was identified from hexanic extracts of strain Asp59, while naphthomycin from ethyl acetate extracts of strain Asp58. These compounds showed insecticidal activity against first instars of <i>Spodoptera frugiperda</i> (Asp59: LC₅₀ = 10.82 µg/µL, LC₉₀ = 26.25 µg/µL; Asp58: LC₅₀ = 15.05 µg/µL, LC₉₀ = 38.84 µg/µL). Notably, this is the first report of naphthomycin as an insecticidal compound. The present study suggests that valinomycin and naphthomycin may be a novel biological source for the control of <i>Spodoptera frugiperda</i> in early stages.