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In 1912 Isonicotinic acid hydrazide (INH) was synthesized from ethyl isonicotinate and hydrazine by Meyer and Malley as part of their doctoral work in Prague. In 1945 its antituberculosis properties were discovered when nicotinamide was discovered to have antituberculosis effects. This prompted the testing of other pyridine derivatives for their antimycobacterial effects.

Investigations at pharmaceutical companies Hoffman La Roche, Farbenfabriken Bayer, and Squibb Institute for Medical Research each independently discovered INH as an antituberculosis agent. The first clinical trial began in 1951 at Sea View Hospital in Staten Island, New York and was reported to the public in 1952.

Tuberculosis became a focus of national attention in 1959 when it became the subject of the Arden House Conference where national efforts were refocused to eliminate tuberculosis from America. The meeting reccomended nationwide chemotherapy for all persons with active tuberculosis and support for a preventative treatment program for those infected with latent bacteria. The discovery of Isoniazid as an appropriate treatment made this goal seem reachable.

Earlier, streptomycin was the main treatment used against tuberculosis, but as a result of streptomycin's use as a single line treatment, the development of drug resistance lowered its effectiveness. The discovery of Isoniazid established a combined drug regimen, neccesary for effective treatment against tuberculosis.

In 1967 the American Thoracic Society reccomended that everyone with a positive tuberculin skin test receive isoniazid chemotherapy to prevent disease progression.