Peptide derivatives corresponding to the carboxy terminal sequen

Details Peptide derivatives corresponding to the carboxy terminal sequen Peptide derivatives corresponding to the carboxy terminal sequen

1994-10-20

1997-11-11

Tsang, Ceclia J.

Chemistry: natural resins or derivatives; peptides or proteins;

Peptides of 3 to 100 amino acid residues

11 to 14 amino acid residues in defined sequence

530328, 530332, 530345, 435 13, 435214, C07K 706, C12Q 156

Patent

active

056865643

DESCRIPTION:

BRIEF SUMMARY
This application is a 371 of PCT/EP93/00908, filed 15 Apr. 1993.
The invention relates to novel peptide derivatives which have activity as enzyme inhibitors, in particular as inhibitors of thrombin. The invention relates also to methods for the production of such enzyme inhibitors, to pharmaceutical compositions containing the latter and to their use as anticoagulant agents.
Accordingly, the present invention provides a compound of the formula I ##STR2## in which R.sup.1 and R.sup.2 are hydrogen, C.sub.1 -C.sub.4 alkyl or are linked to form C.sub.3 -C.sub.7 cycloalkyl and R.sup.3, R.sup.4 and R.sup.5 are the same or different and are hydrogen, C.sub.1 -C.sub.4 alkyl, hydroxy, OR.sup.6, SR.sup.6, halogen, NR.sup.7 R.sup.8, NO.sub.2, CN, CONR.sup.7 R.sup.8 or CO.sub.2 R.sup.9 wherein R.sup.6 is C.sub.1 -C.sub.4 alkyl or C.sub.7 -C.sub.10 aralkyl and R.sup.7, R.sup.8 and R.sup.9 are the same or different and are hydrogen, C.sub.1 -C.sub.4 alkyl or C.sub.7 -C.sub.10 aralkyl or R.sup.7 and R.sup.8 together with the nitrogen atom to which they are bound form 5 or 6 membered azacycloalkyl NH--C(.dbd.NH)--NH.sub.2)--CO!; X is methine CH or nitrogen; n is an integer from 0 to 7; L is a peptide linker, and H is the carboxy terminal end of hirudin, and salts thereof.
Within the scope of the present description, the definitions used hereinbefore and hereinafter have preferably the following meanings:
C.sub.1 -C.sub.4 Alkyl is, for example, corresponding unbranched alkyl such as ethyl, n-propyl or, especially, methyl. C.sub.3 -C.sub.7 Cycloalkyl is, for example, cyclopentyl or cyclohexyl. Halogen is, for example, fluoro, chloro or bromo. C.sub.7 --C.sub.10 Aralkyl is, for example, phenyl-C.sub.1 -C.sub.4 -alkyl such as 2-phenylethyl or, in particular, benzyl. 5 or 6 membered azacycloalkyl is, for example, pyrrolidyl or piperidyl while 5 or 6 membered oxazacycloalkyl is especially morpholyl.
The peptide linker L comprises 3 to 7 amino acids which may be naturally occurring, more especially genetically encoded, amino acids or synthetic amino acids. Because the role of this portion of the molecule is to provide a bridge between the "N-terminal" aryl moiety and the C-terminal hirudin sequence the length rather than the structure of the linker is of importance. Therefore, the choice of the amino acids to be included into the linker is not crucial. However, in order to avoid any undesired counteraction between the side chain of linker amino acids and amino acids present in the C-terminal hirudin portion of the molecules it is preferred to predominantly (or exclusively) choose neutral amino acids (i.e. amino acids which do not include strongly polar functional groups in their side chains such as carboxy, amino or guanidino groups) for the construction of the linker L. It is also preferred to select amino acids which are devoid of bulky side chains. Examples of amino acids which are useful as part of the linker L are naturally occurring or synthetic .omega.-amino carboxylic acids such as glycine, .beta.-alanine, 4-amino butyric acid, 5-amino pentanoic acid, 6-amino hexanoic acid, 11-amino undecanoic acid, and the like, furthermore other neutral amino acids such as alanine, serine, threonine, glutamine, asparagine, phenylglycine, phenylalanine, and the like. It is also possible to include few, especially one, polar amino acids in the peptide chain of the linker L, such as aspartic acid, glutamic acid, lysine, histidine and the like. The linker L may also include at its C-terminal part one amino acid or a few, such as two or three, amino acids which in the hirudin molecule from which the C-terminal radical H is taken precede said C-terminal part. Especially preferred are linkers L which consist of one or more .omega.-amino carboxylic acids, in particular glycine, 4-amino-butyric acid or 6-amino-caproic acid, and, optionally, one or more, especially one, neutral amino acids, e.g. asparagine. The amino acids in the linker L are linked to each other by means of carboxy amide bonds (peptide bonds) as is the first ("N-terminal") amin

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