Hoechst Aktiengesellschaft

Michael Dorschug

Rainer Obermeier

Johannes Meiwes

Rudiger Marquardt

Gerhard Seipke

Klaus-Peter Koller

Ranier Obermeier

New insulin derivatives of the formula II with an iso-electric point between 5 and 8.5, with improved stability in weakly acid aqueous medium and with a special action profile, and the physiologically tolerated salts of these insulin derivatives, for the treatment of diabetes mellitus; formula II is: ##STR1## in which R.sup.1 denotes H or H-Phe,

A.sup.21, B.sup.30, modified insulin derivatives having an altered

A process is described to specifically hydrolyze the amino acid chain of preproinsulin analogs to give the corresponding insulins. The hydrolysis is catalyzed by clostripain and, if necessary, carboxypeptidase B.

Clostripain catalyzed hydrolysis of preproinsulin analogs into

New insulin derivatives, the use thereof, and a pharmaceutical composition containing them Insulin derivatives having an isoelectric point between 5 and 8.5, or physiologically tolerated salts thereof, of the Formula II ##STR1## in which: R.sup.1 at position B1 denotes H or H-Phe;

A.sup.21 -, B.sup.30 - modified insulin derivatives having an altered

The invention relates to a method for the preparation of polypeptides or proteins by enzymatic cleavage of the oligo- or polyglycine sequence of a fusion protein using an endoprotease.

Method for the selective cleavage of fusion proteins with lysostaphin

Pharmaceutical formulations for the treatment of diabetes mellitus are disclosed, wherein the pharmaceutical formulations contain at least one insulin derivative modified with a base in position B31 of the insulin B chain and having an isoelectric point between 5.8 and 8.5 and/or at least one of its physiologically tolerated salts in a pharmaceutically acceptable excipient and a relatively high zinc ion content in the range from above 1 .mu.g to about 200 .mu.g of zinc/IU. The preferred insulin derivative modified with a base for this pharmaceutical formulation is human insulin-B31-Arg-OH and human insulin-B31-Arg-B32-Arg-OH. The formulation is used for the treatment of diabetes mellitus; it has a particularly favorable action profile.

Pharmaceutical formulation for the treatment of diabetes mellitus

Basic proteins are isolated from protein mixtures which contain such basic proteins and which are obtained by enzymatic clevage of proinsulin and/or its derivatives of natural, semisynthetic or genetic engineering origin by ion exchanger chromatography on strongly acid cation exchangers.

Process for isolating basic proteins from protein mixtures

Mixed crystals composed of (a) unmodified insulin, des-Phe-B1 insulin, des-Thr-B30 (human) or des-Ala-B30 (pork, beef) insulin and (b) least one insulin having a basic modification at the C-terminal end of the B chain, preferably composed of human insulin and Arg-B31 human insulin and/or Arg.sub.2 -(B31-32) human insulin. These mixed crystals are obtained by crystallization from aqueous solution within a narrow pH range (from about 5.5 to about 6.9). The mixed crystals show a specific moderately prolonged profile and are suitable for the treatment of diabetes mellitus.

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Bochum, Germany

Michael Dorschug