Peptide linkage unit

Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Peptide containing doai

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530323, 530326, 530327, 530328, 530329, 530330, 562 17, 562 18, 930 30, A61K 3803, C07K 400, C07K 502, C07K 702

Patent

active

055631215

DESCRIPTION:

BRIEF SUMMARY
DESCRIPTION

1. Technical Field
The present invention relates to linkage units for joining peptide sequences and to the use of such linkage units for forming peptides and pseudopeptides, including pseudopeptides that inhibit aspartic proteinase enzymes. More particularly, the invention relates to pseudopeptides that include a phosphinate methylene ammonium linkage (exploding transition state analog) in place of an amide bond at the position in a peptide sequence that is cleaved by aspartic proteinase enzymes.
2. Background Art
Peptide linkage units are employed in the construction of peptides. In the simplest case, naturally occurring L-amino acids serve as peptide linkage units. However, various unnatural L-amino acids and a wide variety of D-amino acids may also serve as peptide linkage units. The peptide linkage unit need not employ a peptide bond to form the linkage. A number of pseudopeptides are disclosed and employed in the prior art as peptide linkage units.
A peptide linkage unit may be employed to link two peptide sequences or may be employed at the terminal end of a peptide. The peptides lying on either side of a peptide linkage unit may be functionally distinct.
Peptide linkage units are employed in the construction of synthetic proteinase inhibitors. Many proteinase substrates include a proteinase binding or recognition region which flanks either side of the cleavage site. Accordingly, one class of synthetic proteinase inhibitor employs peptides having amino acid sequence homology with the binding or recognition regions of known proteinase substrates. If two such peptides are employed in a synthetic proteinase inhibitor, i.e. peptides having a sequence homology with the binding or recognition region flanking either side of a cleavage site, a peptide linkage unit may then be employed to link the two peptides together. In this instance, the peptide linkage unit will be positioned at or near the cleavage site. Peptide linkage units which are resistant to proteolysis and which bind tightly to the active site of the proteinase are particularly useful in the construction of synthetic proteinase inhibitors.
Aspartic proteinase enzymes (EC 3.4.23) are a family of related enzymes that cleave (hydrolyze) protein and polypeptide chains. These enzymes have isoelectric points on the acid side of neutrality and molecule masses ranging from 35,000-45,000 Daltons (D) for fungal enzymes and about 35,500 D for pepsin.
Exemplary enzymes of this class include pepsin that is a mammalian gastric proteinase, cathepsin D that is the intracellular aspartic proteinase of the lysosomal system and whose level has been positively correlated with recurring breast cancers [Tandon et al., N. Eng. J. Med., 322:297 (1990)] and with amyloid formation in Alzheimer's plagues [Cataldo et al., Brain Res., 513:181 (1991)], renin that regulates blood pressure by its cleavage of angiotensinogen to form angiotensin I, and chymosin (formerly called rennin) that cleaves milk proteins as a first step in cheese making. Penicillopepsin, a microbial enzyme from P. janthinellum is another member of this family, whereas nepenthesin, the digestive proteinase of the pitcher plant is exemplary of the plant aspartic proteinases.
The HIV-1 virus also contains an aspartic protease. That enzyme is known to cleave the p17-p24 region of the Pr55.sup.gag fusion protein. Moore et al., Biochem, Biophys. Res. Comm., 159:420 (1989).
Although the precise mechanism of action of this family of enzymes is not as well known as that of the serine proteinases, it is believed that two aspartic acid groups act with water in the active site to cause hydrolysis of the peptide bond that is hydrolyzed. The hydrolyzed peptide bond is typically between hydrophobic residues. A covalent intermediate is not thought to be formed between this enzyme and its substrate as is the case with the serine proteinase family.
This family of enzymes forms an enzyme-substrate complex as is typical in enzyme-substrate reactions. Binding is often found to be a two-step process even though n

REFERENCES:
patent: 4148624 (1979-04-01), Maier
Bartlett, et al., "Potent Inhibition of Pepsin and Pepicillo-Pepsin by Phosphorus-Containing Peptide Analogs", J. Org. Chem., 55:6268-6274 (1990).
Huff, et al., "HIV. Protease: A Novel Chemotherapeutic Target for AIDS", J. Med. Chem, 34: 2305-2314 (1991).

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