Purpose: To formulate novel topical antibacterial gel containing Doxycycline Hyclate, Tea tree oil and Neem oil to evaluate their antibacterial activity. Methods: Formulation excipients were selected using Drug-excipients compatibility study. Prepared gel was subjected to various evaluation parameters like pH, viscosity, spreadability, homogeneity, drug content uniformity, in-vitro drug diffusion and antibacterial activity. Results: Prepared topical gel of doxycycline Hyclate was shown pH range 5.3±0.010 to 6.7±0.015, viscosity range 386±28.70 to 680±50.40 cp, spreadability range 18.14±0.98 to 28.13±2.18 g.cm/s and homogeneous. Prepared gel was shown uniformity in drug content which ranges from 94.41±0.61 to 97.44±0.69%, zone of inhibition ranges 13.66±1.69 to 32.00±1.63 mm for Staphylococcus aureus, 04.00±1.63 to 12.33±0.94 mm for Pseudomonas aeruginosa and 11.66±0.94 to 33.33±1.24 mm for Propionibacterium acne. Prepared gel formulations were shown the negligible amount of drug diffusion, not more than 3.22±0.04% was shown the non-fickian type of diffusion through the egg membrane. Conclusion: Formulation F8 was shown better antibacterial activity as compared to its other formulations. As the concentration of neem oil and tea tree oil was increased from 0.5 to 3% and 0.5 to 2% respectively in the gel, antibacterial activity was synergistically improved against Staphylococcus aureous, Pseudomonas aeruginosa and Propionibacterium acne. The prepared gel formulation was found useful for topical application due to its negligible diffusion, good spreadability, neutral pH, low viscosity and no irritation on human skin.