NKS-01 can be obtained by four similar ways: 1) The microbial hydroxylation of 4-androstene-3,17-dione (I) by a culture of Acremonium strictum NN-106 (FERM P-9143) gives 6beta,14alpha-dihydroxy-4-androstene-3,17-dione (II), which is then oxidized, either photochemically with oxygen in CHCl3 under sunlight , with MgO2 at room temperature (2) or with CrO3/H2SO4. 2) The selective microbial hydroxylation of (I) with a culture of Acremonium strictum NN-106 gives 14alpha-hydroxy-4-androstene-3,17-dione (III), which by alkylation of its enolic form is converted into 3-alkoxy-14alpha-hydroxy-3,5-androstadien-17-one (IV) (R = C1-3 alkyl). Finally, (IV) is oxidized with Jones' reagent (CrO3/H2SO4). 3) By oxidation of 3beta,14alpha-dihydroxy-5-androsten-17-one (V) with Jones' reagent (CrO3/H2SO4) in acetone or with K2Cr2O7 in acetic acid. 4) The microbial hydroxylation of (I) by a microorganism of the genus Myrothecium, such as Myrothecium sp. NK-928521 or natural or artificial mutants, gives (II), which is then oxidized with activated MgO2 in CHCl3 at room temperature.