<p>Specialized and sophisticated enterprises, as the leaders among small- and medium-sized enterprises, have become the backbone of promoting China’s economic and social development and enhancing the international status of China’s value chain. The study aims to explore the spatial distribution and agglomeration hotspots of “Little Giant” Enterprises at both the national- and prefecture-level city scales and also explores where they are and why agglomerate. The study selects five batches of China’s national-level specialized and sophisticated “Little Giant” Enterprises announced by the Ministry of Industry and Information Technology from May 2019 to July 2023, totalling 12,950 China’s specialized and sophisticated “Little Giant” Enterprises as the sample data. The study employed ArcGIS 10.7, a geographic information system platform to transform the enterprise coordinates into enterprise spatial point data files. The study result reveals that the spatial layout of “Little Giant” Enterprises shows a pattern of density in the east and clustered in the key cities of urban agglomeration. The spatial distribution characteristics of “Little Giant” Enterprises are influenced by the regional innovation level, external financial support, policy support, road network density, city-grade infrastructure, industrial structure, and social credit. The novelty of the study lies in realizing the areas where “Little Giant” companies are most focused and the elements that are operating in the geographic focus of these companies in China. This research offers a policy formulation that fosters strategic clustering for the development of Little Giant companies across the country.</p>