Methods and compositions for treating neurodegenerative disorders and alzheimer's disease and improving normal memory
View Patent ↗The disclosure relates generally to neurodegenerative disorders and more specifically to a group of presenilin/G-protein/c-src binding polypeptides and methods of use for modulating signaling and progression of Alzheimer's disease.
1. An isolated polypeptide consisting essentially of an amino acid sequence from the N-terminal fragment of PS-1 (SEQ ID NO:2), wherein the fragment is selected from the group consisting of:
(i) RQVVEQDEEEDEELT (SEQ ID NO:16),
(ii) SEQ ID NO:16 consisting of 1-2 conservative amino acid substitutions, wherein the polypeptide inhibits cell-cell interaction, inhibits Aβ production or binds to a β-APP and wherein the polypeptide retains the sequence of SEQ ID NO:5 (DEEEDEEL),
(iii) the sequence of (i) or (ii) further including 1-10 additional amino acids at the N- or C-terminus, wherein the polypeptide inhibits cell-cell interaction, inhibits Aβ production or binds to a β-APP,
(iv) DEEEDEELTLKYGAK (SEQ ID NO:17),
(v) SEQ ID NO:17 consisting of 1-2 conservative amino acid substitutions, wherein the polypeptide inhibits cell-cell interaction, inhibits Aβ production or binds to a β-APP and wherein the polypeptide retains the sequence of SEQ ID NO:5 (DEEEDEEL),
(vi) a sequence of (iv) or (v) further including 1-10 additional amino acids at the N- or C-terminus, wherein the polypeptide inhibits cell-cell interaction, inhibits Aβ production or binds to a β-APP, and
(vii) any of the foregoing polypeptides comprising an unnatural amino acid or D-amino acid wherein the polypeptide inhibits cell-cell interaction, inhibits Aβ production or binds to a β-APP.
2. The polypeptide of claim 1 , wherein the isolated polypeptide consists of the amino acid sequence RQVVEQDEEEDEELT (SEQ ID NO:16).
3. The polypeptide of claim 1 , wherein the isolated polypeptide consists of the amino acid sequence DEEEDEELTLKYGAK (SEQ ID NO:17).
4. An isolated polynucleotide consisting essentially of a nucleotide sequence encoding a polypeptide consisting essential of an amino acid sequence from the N-terminal fragment of PS-1 (SEQ ID NO:2), wherein the fragment is selected from the group consisting of:
(i) RQVVEQDEEEDEELT (SEQ ID NO:16),
(ii) SEQ ID NO:16 consisting of 1-2 conservative amino acid substitutions, wherein the polypeptide inhibits cell-cell interaction, inhibits Aβ production or binds to a β-APP and wherein the polypeptide retains the sequence of SEQ ID NO:5 (DEEEDEEL),
(iii) the sequence of (i) or (ii) further including 1-10 additional amino acids at the N- or C-terminus, wherein the polypeptide inhibits cell-cell interaction, inhibits Aβ production or binds to a β-APP,
(iv) DEEEDEELTLKYGAK (SEQ ID NO:17),
(v) SEQ ID NO:17 consisting of 1-2 conservative amino acid substitutions, wherein the polypeptide inhibits cell-cell interaction, inhibits Aβ production or binds to a β-APP and wherein the polypeptide retains the sequence of SEQ ID NO:5 (DEEEDEEL),
(vi) a sequence of (iv) or (v) further including 1-10 additional amino acids at the N- or C-terminus, wherein the polypeptide inhibits cell-cell interaction, inhibits Aβ production or binds to a β-APP, and
(vii) any of the foregoing polypeptides comprising an unnatural amino acid or D-amino acid wherein the polypeptide inhibits cell-cell interaction, inhibits Aβ production or binds to a β-APP.
5. An expression vector comprising the polynucleotide of claim 4 .
6. A recombinant host cell comprising the polynucleotide of claim 4 .
7. A method comprising culturing the recombinant host cell of claim 6 under conditions to express the polynucleotide.
8. The method of claim 7 , further comprising isolating the polypeptide expressed from the polynucleotide.
9. An isolated polypeptide produced by the method of claim 8 .
10. A method of inhibiting the production of Aβ comprising contacting mammalian cells expressing a β-amyloid precursor protein (β-APP) with an isolated polypeptide consisting essentially of an amino acid sequence from the N-terminal fragment of PS-1 (SEQ ID NO:2), wherein the polypeptide is selected from the group consisting of:
(i) RQVVEQDEEEDEELT (SEQ ID NO:16),
(ii) SEQ ID NO:16 consisting of 1-2 conservative amino acid substitutions, wherein the polypeptide inhibits Aβ production and wherein the polypeptide retains the sequence of SEQ ID NO:5 (DEEEDEEL),
(iii) the sequence of (i) or (ii) further including 1-10 additional amino acids at the N- or C-terminus, wherein the polypeptide inhibits Aβ production,
(iv) any of the foregoing polypeptides comprising an unnatural amino acid or D-amino acid wherein the polypeptide inhibits Aβ production, and
(v) combinations of any of the foregoing.
11. The method of claim 10 , wherein the isolated polypeptide consists of the amino acid sequence RQVVEQDEEEDEELT (SEQ ID NO:16).
12. A method of inhibiting interaction of β-amyloid precursor protein (β-APP) and presenilin-1 (PS-1) comprising contacting mammalian cells expressing β-APP and/or PS-1 with an isolated polypeptide consisting essentially of an amino acid sequence from the N-terminal fragment of PS-1 (SEQ ID NO:2), wherein the fragment is selected from the group consisting of:
(i) RQVVEQDEEEDEELT (SEQ ID NO:16),
(ii) SEQ ID NO:16 consisting of 1-2 conservative amino acid substitutions, wherein the polypeptide inhibits interaction between β-APP and PS-1 and wherein the polypeptide retains the sequence of SEQ ID NO:5 (DEEEDEEL),
(iii) the sequence of (i) or (ii) further including 1-10 additional amino acids at the N- or C-terminus, wherein the polypeptide inhibits interaction between β-APP and PS-1,
(iv) any of the foregoing polypeptides comprising an unnatural amino acid or D-amino acid wherein the polypeptide inhibits interaction between β-APP and PS-1, and
(v) combinations of any of the foregoing.
13. The method of claim 12 , wherein the isolated polypeptide consists of the amino acid sequence RQVVEQDEEEDEELT (SEQ ID NO:16).
14. A pharmaceutical composition comprising an isolated polypeptide in a pharmaceutically acceptable carrier, wherein the polypeptide is selected from the group consisting of:
(i) RQVVEQDEEEDEELT (SEQ ID NO:16),
(ii) SEQ ID NO:16 consisting of 1-2 conservative amino acid substitutions and wherein the polypeptide retains the sequence of SEQ ID NO:5 (DEEEDEEL),
(iii) the sequence consisting of (i) or (ii) further including 1-10 additional amino acids at the N- and/or C-terminal end(s), and
(iv) any of the foregoing polypeptides comprising an unnatural amino acid or D-amino acid,
wherein the isolated polypeptide inhibits production of Aβ in a cell.
15. The pharmaceutical composition of claim 14 , wherein the polypeptide has the sequence RQVVEQDEEEDEELT (SEQ ID NO:16).
16. The pharmaceutical composition of claim 14 , wherein the composition includes a sterile aqueous solution.
17. The pharmaceutical composition of claim 14 , wherein the composition is in a solvent-free form.