Chimeric proteins
AssigneeChiron Corporation
InventorsMichael A. Innis, Abla A. Creasey
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Abstract
Chimeric proteins possessing Kunitz-type domain 1 of TFPI-2 and Kunitz-type domain 2 of TFPI are disclosed, as are muteins of TFPI and TFPI-2. Nucleic acid sequences, expression vectors and transformed host cells encoding and capable of producing the disclosed chimeric proteins and muteins are also disclosed. Finally, methods for prevention and treatment of septic shock using the chimeric proteins and muteins are disclosed.
Claims38
A chimeric protein comprising:
- (a) a Kunitz-type domain 1 of Tissue Factor Pathway Inhibitor-2 (TFPI-2) or a mutein thereof, wherein the mutein is selected from the group consisting of:
- (i) a Kunitz-type domain 1 of TFPI-2 comprising 1-5 conservative amino acid substitutions that do not substantially change the conformation of TFPI-2;
- (ii) a Kunitz-type domain 1 of TFPI-2 comprising an amino acid substitution that eliminates an N-linked glycosylation site;
- (iii) a Kunitz-type domain 1 of TFPI-2 which has 1-5 anino acid substitutions that change a residue of TFPI-2 to a corresponding residue of TFPI;
- and
- (iv) a Kunitz-type domain 1 of TFPI-2 that comprises an amino acid substitution in the P1 reactive site and
- (b) a Kunitz-type domain 2 of Tissue Factor Pathway Inhibitor (TFPI) or a mutein thereof, wherein the mutein is selected from the group consisting of:
- (i) a Kunitz-type domain 2 of TFPI comprising 1-5 conservative amino acid substitutions that do not substantially change the conformation of TFPI-2;
- (ii) a Kunitz-type domain 2 of TFPI comprising an amino acid substitution that eliminates an N-linked glycosylation site;
- (iii) a Kunitz-type domain 2 of TFPI which has 1-5 amino acid substitutions that change a residue of TFPI to a corresponding residue of TFPI-2;
- and
- (iv) a Kunitz-type domain 2 of TFPI that comprises an amino acid substitution in the P1 reactive site;
- or
- (c) a Kunitz-type domain 2 of TFPI-2 or a mutein thereof, wherein the mutein is selected from the group consisting of:
- (i) a Kunitz-type domain 2 of TFPI-2 comprising 1-5 conservative amino acid substitutions that do not substantially change the conformation of TFPI-2;
- (ii) a Kunitz-type domain 2 of TFPI-2 comprising an amino acid substitution that eliminates an N-linked glycosylation site;
- (iii) a Kunitz-type domain 2 of TFPI-2 which has 1-5 amino acid substitutions that change a residue of TFPI-2 to a corresponding residue of TFPI;
- and
- (iv) a Kunitz-type domain 2 of TFPI-2 that comprises an amino acid substitution in the P1 reactive site;
- and
- (d) a Kunitz-type domain 1 of TFPI or a mutein thereof, wherein the mutein is selected from the group consisting of:
- (i) a Kunitz-type domain 1 of TFPI comprising 1-5 conservative amino acid substitutions that do not substantially change the conformation of TFPI;
- (ii) a Kunitz-type domain 1 of TFPI comprising an amino acid substitution that eliminates an N-linked glycosylation site;
- (iii) a Kunitz-type domain 1 of TFPI which has 1-5 amino acid substitutions that change a residue of TFPI to a corresponding residue of TFPI-2;
- and
- (iv) a Kunitz-type domain 1 of TFPI that comprises an amino acid substitution in the P1 reactive site,
- wherein the chimeric protein binds and inhibits factor VIIa/tissue factor complex and binds to and inhibits factor Xa.
A chimeric protein, wherein the primary amino acid sequence of the chimeric protein is SEQ ID NO:
- 19.
A chimeric protein, wherein the chimeric protein comprises first and second amino acid sequences, said first amino acid sequence comprising SEQ ID NO:
- 19 and said second amino acid sequence selected from the group consisting of:
- (a) SEQ ID NO:
- 7;
- (b) SEQ ID NO:
- 8;
- (c) SEQ ID NO:
- 10;
- (d) SEQ ID NO:
- 11;
- (e) SEQ ID NO:
- 12;
- (f) SEQ ID NO:
- 13;
- (g) SEQ ID NO:
- 14;
- (h) SEQ ID NO:
- 15;
- (i) SEQ ID NO:
- 16;
- (j) SEQ ID NO:
- 17;
- and
- (k) SEQ ID NO:
- 18.
Dependencies are inferred from the claim text.