Claims
- 1. An inhibitor of catalytically active memapsin 2 which binds to the active site of the memapsin 2 defined by the presence of two catalytic aspartic residues and substrate binding cleft, the inhibitor having an Ki of less than or equal to 10−7 M.
- 2. The inhibitor of claim 1, comprising an isostere of the active site of memapsin 2.
- 3. The inhibitor of claim 2, comprising a molecule having the general form
X-L4-P4-L3-P3-L2-P2-L1-P1-L0-P1′-L1′-P2′-L2′-P3′-L3′-P4′-L4′-Y, wherein Px represents the substrate specificity position relative to the cleavage site which is represented by an -L0-, and Lx represent the linking regions between each substrate specificity position, Px and wherein L0 is a non-hydrolyzable bond and P1′ is —R1CR3—, wherein R1 is a group smaller than CH2OH (side chain of serine), and at least two other P positions are a hydrophobic group.
- 4. The inhibitor of claim 1 having a Ki of less than or equal to 10−6 M.
- 5. The inhibitor of claim 1 having a Ki of less than or equal to 2 nM.
- 6. The inhibitor of claim 1 having a Ki of less than or equal to 1 nM.
- 7. The inhibitor of claim 1 having a root mean square difference of less than or equal to 0.5 Å for the side chain and backbone atoms for amino acids 28-441 of SEQ ID NO: 2.
- 8. The inhibitor of claim 1 which is permeable to the blood brain barrier.
- 9. The inhibitor of claim 1 which is less than 900 daltons in molecular weight.
- 10. The inhibitor of claim 1 which blocks cleavage by memapsin 2 under physiological conditions.
- 11. The inhibitor of claim 1 having a Ki of less than or equal to 10−6 M.
- 12. A compound selected from the group consisting of MMI-005, MMI-012, MMI-017, MMI-018, MMI-025, MMI-026, MMI-037, MMI-039, MMI-040, MMI-065, MMI-066, MMI-070, and MMI-071.
- 13. A compound selected from the group consisting of MMI-012, MMI-017, MMI-018, MMI-026, MMI-037, MMI-039, MMI-040, MMI-070 and MMI-071.
- 14. A method for treating a patient to decrease the likelihood of developing or the progression of Alzheimer's disease comprising administering to the individual an effective amount of an inhibitor of memapsin 2 selected from the group consisting of MMI-005, MMI-012, MMI-017, MMI-018, MMI-025, MMI-026, MMI-037, MMI-039, MMI-040, MMI-065, MMI-070 and MMI-071.
- 15. The method of claim 14, wherein the inhibitor is administered orally.
- 16. The method of claim 14, wherein the inhibitor blocks cleavage of APP.
- 17. A method of determining the substrate side-chain preference in memapsin 2 sub-sites, comprising the steps of:
a) reacting a mixture of memapsin 2 substrates with memapsin 2; and b) determining the sub-site preference of memapsin 2 by determining relative initial hydrolysis rates of the mixture of memapsin 2 substrates.
- 18. The method of claim 17, wherein the initial hydrolysis rate is determined by using a MALDI-TOF/MS device wherein the ion intensities from the MALDI-TOF/MS measurements are used for quantification of relative amounts of the substrates and the products formed thereof by hydrolysis of the substrates.
- 19. A method of determining the substrate side-chain preference in memapsin 2 sub-sites, comprising the steps of:
a) preparing a combinatorial library of memapsin 2 inhibitors wherein the inhibitors comprise a base sequence taken from OM99-2; b) probing the library of inhibitors with memapsin 2 wherein the memapsin 2 may bind one or a plurality of inhibitors to generate one or a plurality of bound memapsin 2; and c) detecting the bound memapsin 2 with an antibody raised to memapsin 2 and an alkaline phosphatase conjugated secondary antibody.
- 20. The method of claim 19, wherein the inhibitors are immobilized on beads.
- 21. The method of claim 19, wherein the base sequence is EVNL*AAEF and wherein the P3, P2, P2′, and P3′ sub-sites of memapsin 2 are randomized.
- 22. A method of treating a human suffering from Alzheimer's disease comprising administering to the human an inhibitor of catalytically active memapsin 2 which binds to the active site of the memapsin 2 defined by the presence of two catalytic aspartic residues and substrate binding cleft, the inhibitor having an Ki of less than or equal to 10−7 M.
- 23. A method of treating a human suffering from Alzheimer's disease comprising administering to the human an inhibitor of catalytically active memapsin 2 which binds to the active site of the memapsin 2 defined by the presence of two catalytic aspartic residues and substrate binding cleft, the inhibitor having an Ki of less than or equal to 10−7 M, wherein the inhibitor has a root mean square difference of less than or equal to 0.5 Å for the side chain and backbone atoms for amino acids 28-441 of SEQ ID NO: 2.
- 24. A method of treating a human suffering from Alzheimer's disease comprising administering to the human a compound selected from the group consisting of MMI-012, MMI-017, MMI-018, MMI-026, MMI-037, MMI-039, MMI-040, MMI-070 and MMI-071.
RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional Application No. 60/275,756, filed on Mar. 14, 2001 and 60/258,705 filed on Dec. 28, 2000, the entire teachings of both of which are incorporated by reference in their entirety.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60275756 |
Mar 2001 |
US |
|
60258705 |
Dec 2000 |
US |