Nova Patents
EP0339942A2

Aprotinin analogues and process for the production thereof.

Abstract

Novel aprotinin analogues having a selected inhibition profile against serine proteases.

EP0339942A2, drawing sheet 1
Sheet 1 of 11

Term

Term ended

Projected expiry passed 25 April 2009, 17.4 years ago.

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23 claims: 11 independent, 12 dependent

  1. 1
    Aprotinin analogues having the formula X₁-Asp-Phe-Cys-Leu-Glu-Pro-Pro-Tyr-Thr-X₂-X₃-X₄-X₅-X₆-X₇-­X₈-X₉ Arg-Tyr-Phe-Tyr-Asn-Ala-Lys-Ala-Gly-Leu-Cys-Gln-Thr-­Phe-Val-Tyr-Gly-Gly-X₁₀-Arg-Ala-X₁₁-X₁₂-Asn-Asn-Phe-Lys-­Ser-Ala-Glu-Asp-Cys-Met-Arg-Thr-Cys-Gly-Gly-Ala in which X₁ means Arg-Pro, Pro or hydrogen, X₄ and X₁₀ are both Cys or both Ala or both another naturally occuring amino acid residue, X₂ and X₃ are independently any naturally occuring amino acid residue;X₅ is Lys, Arg, Val, Thr, Ile, Leu, Phe, Gly, Ser, Met, Trp, Tyr or Ala with the proviso that when X₅ is different from Lys then at least one of X₂ to X₄ and X₆ to X₉ is different from the amino acid residue at the corresponding position in native aprotinin;X₆ is Ala or Gly;X₇ is any naturally ocurring amino acid residue;X₈ is Ile, Leu, Met, Val or Phe;X₉ is any naturally occuring amino acid residue;X₁₁ is any naturally occuring amino acid residue;X₁₂ is Lys, Arg or Ser at least one of the amino acid residues X₂ to X₉ being different from the corresponding amino acid residue in native aprotinin.
  2. 4
    Aprotinin analogues according to any of the preceding claims, wherein X₃ is Pro.
  3. 5
    Aprotinin analogues according to any of the preceding claims, wherein X₄ and X₁₀ are Cys.
  4. 6
    Aprotinin analogues according to any of the preceding claims wherein X₅ is Lys, Arg, Val, Thr, Ile, Leu, Phe, Gly, Ser, Met, Trp, Tyr or Ala, preferably Lys or Arg with the proviso that when X₅ is different from Lys, then at least one of X₂ to X₄ and X₆ to X₉, preferably X₆ to X₉ is different from the amino acid residue at the corresponding position in native aprotinin.
  5. 7
    Aprotinin analogues according to any of the preceding claims, wherein X₆ is Ala or Gly, preferably Ala.
  6. 8
    Aprotinin analogues according to any of the preceding claims, wherein X₇ is Ala or Gly, preferably Ala.
  7. 9
    Aprotinin analogues according to any of the preceding claims wherein X₈ is Ile, Leu, Met, Val or Phe, preferably Ile.
  8. 10
    Aprotinin analogues according to any of the preceding claims wherein X₉ is Ile or Glu, preferably Ile.
  9. 11
    Aprotinin analogues according to any of the preceding claims wherein X₁₁ is Lys or Arg, preferably Lys.
  10. 12
    Aprotinin analogues according to any of the preceding claims wherein X₁₂ is Lys, Arg o Ser, preferably Arg or Ser.
  11. 23
    A process for producing aprotinin analogues according to the general formula (I):X₁-Asp-Phe-Cys-Leu-Glu-Pro-Pro-Tyr-Thr-X₂-X₃-X₄-X₅-X₆-X₇-­X₈-X₉-Arg-Tyr-Phe-Tyr-Asn-Ala-Lys-Ala-Gly-Leu-Cys-Gln-Thr-­Phe-Val-Tyr-Gly-Gly-X₁₀-Arg-Ala-X₁₁-X₁₂-Asn-Asn-Phe-Lys-­Ser-Ala-Glu-Asp-Cys-Met-Arg-Thr-Cys-Gly-Gly-Ala      (I) wherein X₁ is Arg-Pro, Pro or hydrogen;X₂ is any naturally occuring amino acid residue, preferably Gly;X₃ is any naturally occuring amino acid residue, preferably Pro;X₄ and X₁₀ are any naturally occuring amino acid residues, preferably they are both Ala or both Cys;X₅ is Lys, Arg, Val, Thr, Ile, Leu, Phe, Gly, Ser, Met, Trp, Tyr or Ala, preferably Lys or Arg, with the proviso, that when X₅ is different from Lys, then at least one of X₂ to X₄ and X₆ to X₉ is different from the amino acid residue at the corresponding position in native aprotinin;X₆ is Ala or Gly, preferably Ala;X₇ is any naturally occuring amino acid residue, preferably Ala or Gly;X₈ is Ile, Leu, Met, Val or Phe, preferably Ile;X₉ is any naturally occuring amino acid residue, preferably Ile or Glu;X₁₁ is any naturally occuring amino acid residue, preferably Lys or Arg and X₁₂ is Lys, Arg or Ser, preferably Arg or Ser, at least one of the amino acid residues X₂ to X₉, preferably X₅ to X₉, more preferred X₆ to X₉ being different from the corresponding amino acid residue in native protein in yeast comprising culturing a yeast strain containing a replicable expression vector, the vector comprising a gene coding for the aprotinin analogue and DNA sequences that allow for the expression of the aprotinin analogue in a suitable nutrient medium and recovering the expressed aprotinin analogues.
Independent claims11