التفاصيل البيبلوغرافية
| العنوان: |
RECOMBINANTLY MODIFIED PLASMIN |
| Document Number: |
20100047228 |
| تاريخ النشر: |
February 25, 2010 |
| Appl. No: |
12/090782 |
| Application Filed: |
October 18, 2006 |
| مستخلص: |
Methods of using polynucleotides and polypeptides relating to a recombinantly-modified plasmin(ogen) molecule are provided, including methods related to vitrectomy or vitreolysis. The plasmin(ogen) molecule has a single kringel domain N-terminal to the activation site present in the native human plasminogen molecule, and exhibits lysine-binding and significant enzymatic characteristics associated with the native enzyme. |
| Inventors: |
Scuderi, JR., Philip (Chapel Hill, NC, US); Arora, Vikram (Morrisville, NC, US); Hunt, Jennifer A. (Raleigh, NC, US); Novokhatny, Valery (Raleigh, NC, US); Petteway, JR., Stephen R. (Cary, NC, US) |
| Claim: |
1. A method of liquefying a vitreous or a portion thereof, and/or inducing posterior vitreous detachment of an eye of a subject, comprising contacting the vitreous and/or an aqueous humor in the eye of the subject with an effective amount of a composition comprising a polypeptide having a single N-terminal kringle domain homologous to a kringle domain of native human plasminogen; and a C-terminal domain activation site and serine protease domain homologous to the corresponding domains in human plasminogen, wherein the polypeptide binds to immobilized lysine, and wherein the method results in liquefying a vitreous and/or inducing posterior vitreous detachment of the eye of a subject. |
| Claim: |
2. The method of claim 1, wherein the N-terminal kringle domain of the polypeptide is homologous to kringle 1 or kringle 4 of native human plasminogen. |
| Claim: |
3. The method of claim 1, wherein the polypeptide exhibits a fibrinolytic activity that is inhibited by α2-antiplasmin at a rate of inhibition that is at least about 5-fold faster than the rate of inhibition of the fibrinolytic activity of mini-plasmin by α2-antiplasmin. |
| Claim: |
4. The method of claim 3, wherein the rate of inhibition is at least about 10-fold, 20-fold, 30-fold, or 40-fold faster than the rate of inhibition of mini-plasmin. |
| Claim: |
5. The method of claim 1, wherein the immobilized lysine is lysine bound to a solid support matrix selected from the group consisting of lysine-agarose, lysine-hydrogel, lysine-cross-linked agarose. |
| Claim: |
6. The method of claim 5, wherein the immobilized lysine is lysine-cross-linked agarose. |
| Claim: |
7. The method of claim 1, wherein the polypeptide exhibits a lower binding affinity for fibrinogen than the binding affinity for fibrinogen of mini-plasmin. |
| Claim: |
8. The method of claim 1, wherein the polypeptide exhibits higher binding affinity for partially cleaved fibrin than the binding affinity for partially cleaved fibrin of mini-plasmin. |
| Claim: |
9. The method of claim 1, wherein the single kringle domain has at least one residue greater amino acid sequence identity with kringle 1 or kringle 4 of native human plasminogen than with kringle 5 of native human plasminogen, and wherein conservative substitutions of the single kringle region relative to the native sequences of kringles 1 and 4 of human plasminogen are not considered as differing from the native sequences for purposes of the identity comparison with kringle 5. |
| Claim: |
10. The method of claim 9, wherein the polypeptide has an amino acid sequence as shown in SEQ ID NO:2, and conservative substitutions thereof. |
| Claim: |
11. The method of claim 9, wherein the polypeptide has an arginine residue at a relative position analogous to that of position 76 of the amino acid sequence shown in SEQ ID NO:2. |
| Claim: |
12. The method of claim 1, wherein the composition is a liquid solution, and contacting the vitreous and/or the aqueous humor with the composition comprises injecting the liquid solution into the vitreous and/or the aqueous humor. |
| Claim: |
13. The method of claim 1, wherein the effective amount of the polypeptide is in the range of 0.005 mg to 1 mg. |
| Claim: |
14. The method of claim 1, wherein the subject has, or is at risk of developing, a vitreoretinal disease or disorder. |
| Claim: |
15. A method of treating or preventing a vitreoretinal disease or disorder, or a complication of a vitreoretinal disease or disorder, of an eye of a subject, comprising contacting a vitreous and/or an aqueous humor in the eye of the subject with an effective amount of a composition comprising a polypeptide having a single N-terminal kringle domain homologous to a kringle domain of native human plasminogen; and a C-terminal domain activation site and serine protease domain homologous to the corresponding domains in human plasminogen, wherein the polypeptide binds to immobilized lysine, wherein the method results in vitreous liquefaction and/or posterior vitreous detachment in the eye of the subject, thereby treating or preventing the vitreoretinal disease or disorder, or a complication of the vitreoretinal disease or disorder, of the eye of the subject. |
| Claim: |
16. The method of claim 15, wherein the composition is a liquid solution, and contacting the vitreous and/or the aqueous humor with the composition comprises injecting the liquid solution into the vitreous and/or the aqueous humor. |
| Claim: |
17. The method of claim 15 wherein the effective amount of the polypeptide is in the range of 0.005 mg to 1 mg. |
| Claim: |
18. A method of performing a vitrectomy in a subject, comprising contacting a vitreous and/or aqueous humor of an eye of the subject with an effective amount of a composition comprising a polypeptide having a single N-terminal kringle domain homologous to a kringle domain of native human plasminogen; and a C-terminal domain activation site and serine protease domain homologous to the corresponding domains in human plasminogen, wherein the polypeptide binds to immobilized lysine, wherein the contacting is prior to or at the same time as the removal of the vitreous. |
| Claim: |
19. The method of claim 18, wherein the effective amount of the polypeptide is in the range of 0.005 mg to 1 mg. |
| Claim: |
20. The method of claim 18, where the composition is a liquid solution. |
| Claim: |
21. The method of claim 20, wherein contacting the vitreous and/or the aqueous humor with the composition comprises injecting the liquid solution into the vitreous and/or the aqueous humor. |
| Current U.S. Class: |
424/9/464 |
| Current International Class: |
61 |
| رقم الانضمام: |
edspap.20100047228 |
| قاعدة البيانات: |
USPTO Patent Applications |