Therapeutic spalt-like transcription factor 4 (SALL4) peptide

Patent metadata

Field	Value
Publication number	US-10793601-B2
Application number	US-201716095603-A
Country	US
Kind code	B2
Filing date	Apr 28, 2017
Priority date	Apr 28, 2016
Publication date	Oct 6, 2020
Grant date	Oct 6, 2020

Abstract

Isolated peptides and pharmaceutical compositions comprising isolated peptides that bind to retinoblastoma binding protein 4 (RBBp4) and block the Spalt-Like Transcription Factor 4 (SALL4)-RBBp4 interaction are described. Methods of inhibiting binding of SALL4 to RBBp4 and methods of treating a subject having a disorder mediated by a dysregulation of SALL4 are also described.

First claim

We claim:

1. A peptide having less than 12 residues, comprising:
the amino acid sequence set forth in formula I
RRKX 1 X 2 X 3 X 4 X 5 X 6 X 7 (SEQ ID NO: 62),   (I)

wherein X 1 , X 2 , X 3 , X 4 , X 5 , X 6 or X 7 is independently an amino acid with non-polar side chain, polar side chain that is not charged at neutral pH or polar side chain that is positively charged at neutral pH; wherein the peptide does not consist of the amino acid sequence of RRKQAKPQHI (SEQ ID NO: 3); wherein the sequence of amino acids is written from the N-terminus to the C-terminus; and wherein the peptide has an N-terminal acetyl group or protecting group, a C-terminal amine group or protecting group, or both.

2. The peptide of claim 1 , wherein the peptide comprises the amino acid sequence set forth in formula II
A−X 0 RRKX 1 X 2 X 3 X 4 X 5 X 6 X 7 −B (SEQ ID NO: 63),   (II)

wherein X 0 , X 1 , X 2 , X 3 , X 4 , X 5 , X 6 or X 7 is independently an amino acid with non-polar side chain, polar side chain that is not charged at neutral pH or polar side chain that is positively charged at neutral pH;
wherein A is an acetyl group or an N-terminal protecting group;
wherein B is an amine group or a C-terminal protecting group.

3. The peptide of claim 2 , wherein X 0 is an amino acid with a non-polar side chain or a polar side chain that is positively charged at neutral pH.

4. The peptide of claim 1 , wherein the peptide binds to the histone binding protein retinoblastoma binding protein 4 (RBBp4).

5. The peptide of claim 4 , wherein the peptide binds to RBBp4 with dissociation constant of about 1 μM or lower.

6. The peptide of claim 5 , wherein the peptide is

RRKAAKPQHI, RRKFAKPQHI, RRKFAKFQWI,

RRKQKKPQHI, RRKQRKPQHI, RRKQAAPQHI, RRKQAVPQHI,

RRKQALPQHI, RRKQAFPQHI, RRKQARPQHI, RRKQAKFQHI,

RRKQAKPAHI, RRKQAKPEHI, RRKQAKPNHI, RRKQAKPVHI,

RRKQAKPLHI, RRKQAKPQAI, RRKQAKPQFI, RRKQAKPQYI,

RRKQAKPQWI, RRKQAKPQVI, RRKQAKPQHA, RRKQAKPQHV,

RRKQAKPQHL, RRKQAKPQHF, RRKFAKFQWI, RRKHAKPQHI,

ORRKQAKPQHI, HRRKQAKPQHI, RRKQPKPQHI, HRRKQAKPQHI,

URRKQAKPQHI 

or 

ZRRKQAKPQHI, ORRKHPKPQH.

7. The peptide of claim 4 , wherein the molecular weight of the peptide is between about 0.9 to 1.5 kDa.

8. The peptide of claim 1 , wherein the peptide comprises the amino acid sequence set forth in RRKX 1 AKPQHI (SEQ ID NO: 4), wherein X 1 is an amino acid with a non-polar aromatic side chain.

9. The peptide of claim 1 , wherein the peptide comprises the amino acid sequence set forth in RRKQX 2 KPQHI (SEQ ID NO: 5), wherein X 2 is an amino acid with a polar side chain that is positively charged at neutral pH.

10. The peptide of claim 1 , wherein the peptide comprises the amino acid sequence set forth in RRKQAX 3 PQHI (SEQ ID NO: 6), wherein X 3 is an amino acid with a non-polar side chain, or a polar side chain that is positively charged at neutral pH.

11. The peptide of claim 1 , wherein the peptide comprises the amino acid sequence set forth in RRKQAKX 4 QHI (SEQ ID NO: 7), wherein X 4 is an amino acid with a non-polar side chain.

12. The peptide of claim 1 , wherein the peptide comprises the amino acid sequence set forth in RRKQAKPX 5 HI (SEQ ID NO: 8) wherein X 5 is an amino acid with a non-polar side chain, or a polar side chain that is not charged at neutral pH.

13. The peptide of claim 1 , wherein the peptide comprises the amino acid sequence set forth in RRKQAKPQX 6 I (SEQ ID NO: 9) wherein X 6 is an amino acid with a non-polar side chain.

14. The peptide of claim 1 , wherein the peptide comprises the amino acid sequence set forth in RRKQAKPQHX 7 (SEQ ID NO: 10), wherein X 7 is an amino acid with a non-polar side chain.

15. The peptide of claim 1 , wherein the peptide comprises the amino acid sequence set forth in RRKX 1 AKX 4 QX 6 I (SEQ ID NO: 11), wherein X 1 , X 4 or X 6 is independently selected from a group consisting of amino acids with non-polar side chains.

16. The peptide of claim 15 , wherein the peptide comprises the amino acid sequence set forth in X 0 RRKQAKPQHI (SEQ ID NO: 12), wherein X 0 is an amino acid with a non-polar side chain or a polar side chain that is positively charged at neutral pH.

17. A peptide, comprising the peptide of claim 1 and a cell-penetrating peptide attached thereto.

18. The peptide of claim 17 , having a molecular weight between about 2.2 to about 3.8 kDa.

19. The peptide of claim 17 , wherein the cell-penetrating peptide is attached to the N-terminus of the peptide.

20. The peptide of claim 19 , wherein the cell-penetrating peptide comprises an amino acid sequence set forth in RQIKIWFQNRRMKWKK (SEQ ID NO: 2).

21. A peptide comprising a binding peptide and a cell-penetrating peptide attached thereto, wherein the binding peptide:
a) has less than 12 residues;
b) does not consist of RRKQAKPQHI (SEQ ID NO: 3); and
c) comprises the amino acid sequence set forth in formula I
RRKX 1 X 2 X 3 X 4 X 5 X 6 X 7 (SEQ ID NO: 62),   (I)

wherein:
i) X 1 , X 2 , X 3 , X 4 , X 5 , X 6 or X 7 is independently an amino acid with non-polar side chain, polar side chain that is not charged at neutral pH or polar side chain that is positively charged at neutral pH; and
ii) the sequence of amino acids is written from the N-terminus to the C-terminus.

22. A pharmaceutical composition comprising the peptide of claim 21 and a pharmaceutically acceptable carrier.

23. The pharmaceutical composition of claim 22 , wherein the cell-penetrating, peptide comprises an amino acid sequence RQIKIWIFQNRRNIKWKK (SEQ ID NO: 2).

24. The peptide of claim 21 , having a molecular weight of between about 2.2 to about 3.8 kDa.

25. The peptide of claim 21 , wherein the cell-penetrating peptide is attached to the N-terminus of the binding peptide.

26. The peptide of claim 25 , wherein the cell-penetrating peptide comprises an amino acid sequence set forth in RQIKIWFQNRRMKWKK (SEQ ID NO: 2).

27. A method of inhibiting the binding of a Sal-like protein 4 (SALL4) with histone-binding protein retinoblastoma binding protein 4 (RBBp4) in a cell expressing SALL4, comprising contacting the cells with a peptide comprising a binding peptide having less than 12 residues, wherein th

Assignees

• Nat Univ Singapore
• Brigham & Womens Hospital Inc
• Agency Science Tech & Res

Inventors

• Liu Bee Hui
• Tenen Daniel Geoffrey
• Chai Li
• Chia Cheng San Brian
• Poulsen Anders

Classifications

• C07K7/06 Primary

  having 5 to 11 amino acids · CPC title

• C07K2319/033

  containing a motif for targeting to the internal surface of the plasma membrane, e.g. containing a myristoylation motif · CPC title

• C07K7/08

  having 12 to 20 amino acids (gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68) · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title