Florey teve um papel fundamental nos primeiros ensaios clínicos, em 1941, no hospital Radcliffe Infirmary, em Oxford.
Após ensaios em animais, Florey e a sua equipa testaram a penicilina no primeiro paciente humano, a Albert Alexander, um polícia de 43 anos, que desenvolveu uma infeção generalizada, provocada por um corte numa roseira. Durante quatro dias, foram-lhe administradas doses de penicilina, e a melhoria do paciente foi notável logo após as primeiras 24 horas de tratamento. Contudo, o fornecimento de penicilina terminou antes do tratamento ter sido completado, pelo que o paciente acabou por falecer.[2]
Dos cinco pacientes a serem tratados em seguida, quatro recuperaram das suas infeções graças ao tratamento com penicilina.
Para além do prêmio Nobel, Florey foi condecorado com o título de Barão (Barão Florey de Adelaide) no ano de 1965.
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