Human kunitz-type inhibitor with enhanced antifibrinolytic activity
A human Kunitz-type inhibitor polypeptide with enhanced antifibrinolytic activity, methods of making, and methods of use. The novel polypeptide is structurally similar to the KD1 domain of human tissue factor pathway inhibitor-2 (TFPI-2). In another aspect, methods of treating a subject afflicted with cancer or a precancerous condition are described. Generally, the method includes administering to a subject in need of treatment an effective amount of a polypeptide. In some embodiments, the polypeptide comprises a KD1 domain of human TFPI-2. In some embodiments, the polypeptide comprises human TFPI-2, itself. In certain embodiments, the polypeptide is administered in an amount effective to induce apoptosis in tumor cells.
1. A method for treating a subject afflicted with cancer or a precancerous condition, the method comprising:
administering to a subject in need of treatment for cancer or a precancerous condition an effective amount of a polypeptide comprising a KD1 domain consisting of a primary structure that is at least 86% identical to the primary structure of the wild-type KD1 domain of human tissue factor pathway inhibitor-2 (TFPI-2) (SEQ ID NO:2);
wherein the polypeptide does not include a KD2 or KD3 domain of human TFPI-2.
2. The method of claim 1 wherein the KD1 domain of the polypeptide consists of wild-type KD1 (SEQ ID NO:2).
3. The method of claim 1 wherein the KD1 domain of the polypeptide comprises a lysine instead of an arginine at position 24 of the amino acid sequence of SEQ ID NO:2.
4. A method for treating a subject afflicted with cancer or a precancerous condition, the method comprising:
administering to a subject in need of treatment for cancer or a precancerous condition an effective amount of a polypeptide comprising:
a KD1 domain consisting of a primary structure that is at least 86% identical to the primary structure of the wild-type KD1 domain of human TFPI-2; and
a multiply positively charged amino acid sequence disposed at either or both of the N-terminal or C-terminal end of the polypeptide;
wherein the polypeptide does not include a KD2 or KD3 domain of human TFPI-2.
5. The method of claim 4 wherein the KD1 domain of the polypeptide consists of wild-type KD1 (SEQ ID NO:2).
6. The method of claim 4 wherein the KD1 domain of the polypeptide comprises a lysine instead of an arginine at position 24 of the amino acid sequence of SEQ ID NO:2.
7. A method of inducing apoptosis in a cell population, the method comprising:
contacting at least a portion of the cell population with an amount of a polypeptide effective to induce apoptosis in cells of the cell population, wherein the polypeptide comprises a KD1 domain consisting of a primary structure that is at least 86% identical to the primary structure of the wild-type KD1 domain of human tissue factor pathway inhibitor-2 (TFPI-2) (SEQ ID NO:2), and wherein the polypeptide does not include a KD2 or KD3 domain of human TFPI-2.
8. The method of claim 7 wherein the KD1 domain of the polypeptide consists of wild-type KD1 (SEQ ID NO:2).
9. The method of claim 7 wherein the KD1 domain of the polypeptide comprises a lysine instead of an arginine at position 24 of the amino acid sequence of SEQ ID NO:2.
10. A method of inducing apoptosis in a cell population, the method comprising:
contacting at least a portion of the cell population with an amount of a polypeptide effective to induce apoptosis in cells of the cell population, wherein the polypeptide comprises a KD1 domain consisting of a primary structure that is at least 86% identical to the primary structure of the wild-type KD1 domain of human TFPI-2, a multiply positively charged amino acid sequence disposed at either or both of the N-terminal or C-terminal end of the polypeptide, and does not include a KD2 or KD3 domain of human TFPI-2.
11. The method of claim 10 wherein the KD1 domain of the polypeptide consists of wild-type KD1 (SEQ ID NO:2).
12. The method of claim 10 wherein the KD1 domain of the polypeptide comprises a lysine instead of an arginine at position 24 of the amino acid sequence of SEQ ID NO:2.