Merck + Co., Inc.

Patricia M Cameron

Ali Shafiee

Louis Kaplan

David A Boulton

Guadalupe A Limjuco

Jack A Schmidt

Haideh Motamedi

Described is a new enzyme, 31-O-desmethyl-FK-506 O-methyltransferase (FKMT) and its N-terminal amino acid sequence. This enzyme can specifically and preferentially methylate the C-31 hydroxy group in 31-O-desmethyl FK-506/FR-900506 and other 31-O-desmethyl analogs of FK-506/FR-900506 and related compounds such as 31-O-desmethyl immunomycin type molecules, also designated FK-520 and FR-900520 by Fujisawa. The enzyme can be isolated from a FK-506 producing microorganism Streptomyces sp. (Merck Culture Collection No. MA 6858) ATCC No. 55098. Employing the enzyme in its active form, supplemented with Mg.sup.+2 ion, in the presence of the methyl donor, S-adenosyl methionine (SAM), derivatives of 31-O-desmethyl FK-506, 31-O-desmethyl FK-520 and related 31-O-desmethyl compounds may be prepared.

Methlating enzyme from streptomyces MA6858

Human interleukin-1 (IL-1) charged species (pI 6.8, 6.0, 5.4 and 5.2) are separated and purified by a rapid and efficient two or three stage chromatographic process. Interleukin-1 species present in concentrated culture fluid from stimulated human mononuclear cells are separated and partially purified by anion exchange high performance liquid chromatography. The pI 5.4 and 5.2 species in addition are partially purified by HPLC gel filtration chromatography. Each of the individual IL-1 species are then finally purified by reverse phase high performance liquid chromatography. This process results in homogenously pure pI 6.8, pI 5.4 and pI 5.2 species while the pI 6.0 species is isolated in a substantially pure form.

Purification of human interleukin-1 species

Growing community of inventors

Titusville, NJ, United States of America

Patricia M Cameron