Background: Pre-exposure Prophylaxis (PrEP) is a promising intervention to reduce the risk of acquiring HIV among high-risk individuals including men who have unprotected sex with men, HIV discordant couples, heterosexual men or women who have unprotected sex and individuals who inject drugs. PrEP requires adherence to a daily medication and regular clinical monitoring. This needs assessment explored the knowledge and attitudes of physicians towards HIV PrEP to inform the development of a PrEP clinic in Louisville, Kentucky. Methods: Self-administered surveys were distributed to a convenience sample of Family Medicine and Internal Medicine grand round attendees. The Fisher's Exact test was used to assess differences in responses among clinical specialties. Results: Ninety-seven attendees completed the survey (53% primary care physicians, 28% non-primary care physicians, 18% medical students, and 1% no response). While 87% of all respondents support the use of PrEP to prevent HIV among high-risk individuals, only 12% of practicing physicians have prescribed PrEP, citing a lack of clinical knowledge as the main reason. In regards to willingness to prescribe PrEP, there was a significant difference among specialties with 67% of primary care physicians willing to prescribe PrEP, compared to 12% of non-primary care physicians (p < .001). Physicians identified the top three barriers to PrEP administration as high demands on physician time, high costs, and potential patient toxicity. Conclusion: Findings informed the design of an inter-professional clinical model for PrEP involving primary care physicians and pharmacists to reduce barriers to PrEP administration through clinical education and improved inter-professional collaboration.