Anti-inflammatory compounds and uses thereof
The present invention provides anti-inflammatory compounds, pharmaceutical compositions thereof, and methods of use thereof for treating inflammatory disorders. The present invention also provides methods of identifying anti-inflammatory compounds and methods of inhibiting NF-κB-dependent target gene expression in a cell.
1. A method for modulating NF-κB induction in a cell, comprising contacting the cell with an anti-inflammatory compound comprising the structure:
X a —X b ,
wherein X a is a membrane translocation domain; X b is a NF-κB Essential Modulator (NEMO) binding sequence; wherein the anti-inflammatory compound is less than one-hundred amino acids in length; and wherein the anti-inflammatory compound blocks the interaction between one or more IκB protein kinases (IKKs) and NEMO, thereby modulating NF-κB induction in the cell.
2. The method of claim 1 , wherein the anti-inflammatory compound further comprises a modifying group.
3. The method of claim 1 , wherein X 6 comprises the following structure:
(Y) n —X 1 —X 2 —X 3 —X 4 —X 5 —X 6 -(A) m (SEQ ID NO:172)
wherein
n and m are each, independently, 0 or 1;
A and Y each comprises from 1 to about 3 amino acid residues;
X 1 is Leu, Ala, Ile, or nor-leucine (Nle);
X 2 is Asp, Glu, Asn, Gln, homoserine (Hser) or 2-ketopropylalanine (2-ketopropy-A);
X 3 is Trp, Phe, Tyr, 4-biphenyl-alanine (Bpa), homophenylalanine (Hphe), 2-Naphthylalanine (2-NaI), 1-Naphthylalanine (1-NaI), or cycloxexyl-alanine (Cha);
X 4 is Ser, Ala, Glu, Leu, Thr, nor-leucine (Nle), or homoserine (Hser);
X 5 is Trp, His, homophenylalanine (Hphe), 2-Naphthylalanine (2-NaI), 1-Naphthylalanine (1-NaI), O-benzyl serine (SeroBn), or 3-Pyridylalanine (3-Pal); and
X 6 is Leu, Ala, Ile, or nor-leucine (Nle).
4. The method of claim 3 , wherein n is 1 and Y is the sequence Thr-Ala (SEQ ID NO:173).
5. The method of claim 3 , wherein m is 1 and A is the sequence Gln-Thr-Glu (SEQ ID NO:174).
6. The method of claim 1 , wherein said compound comprises a sequence selected from the group consisting of Thr-Ala-Leu-Asp-Trp-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:28), Leu-Asp-Trp-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:29), Thr-Ala-Leu-Asp-Trp-Ser-Trp-Leu (SEQ ID NO:30), Ala-Leu-Asp-Trp-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:31), Leu-Asp-Trp-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:32), Leu-Asp-Trp-Ser-Trp-Leu (SEQ ID NO:33), Thr-Ala-Leu-Asp-Trp-Ser-Trp-Leu-Gln-Thr (SEQ ID NO:34), Thr-Ala-Leu-Asp-Trp-Ser-Trp-Leu-Gln (SEQ ID NO:35), Ala-Leu-Asp-Trp-Ser-Trp-Leu-Gln-Thr (SEQ ID NO:36), Leu-Asp-Trp-Ser-Trp-Leu-Gln (SEQ ID NO:37), Leu-Asp-Trp-Ser-Trp-Leu-Gln-Thr (SEQ ID NO:38), Ala-Asp-Trp-Ser-Trp-Leu (SEQ ID NO:39), Leu-Asp-Trp-Ser-Trp-Ala (SEQ ID NO:40), Ala-Asp-Trp-Ser-Trp-Ala (SEQ ID NO:41), Leu-Asp-Phe-Ser-Trp-Leu (SEQ ID NO:42), Leu-Asp-Tyr-Ser-Trp-Leu (SEQ ID NO:43), Leu-Asp-Trp-Ala-Trp-Leu (SEQ ID NO:44), Leu-Asp-Trp-Glu-Trp-Leu (SEQ ID NO:45), Thr-Ala-Ala-Asp-Trp-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:46), Ala-Asp-Trp-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:47), Thr-Ala-Ala-Asp-Trp-Ser-Trp-Leu (SEQ ID NO:48), Ala-Ala-Asp-Trp-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:49), Ala-Asp-Trp-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:50), Ala-Asp-Trp-Ser-Trp-Leu (SEQ ID NO:51), Thr-Ala-Ala-Asp-Trp-Ser-Trp-Leu-Gln-Thr (SEQ ID NO:52), Thr-Ala-Ala-Asp-Trp-Ser-Trp-Leu-Gln (SEQ ID NO:53), Ala-Ala-Asp-Trp-Ser-Trp-Leu-Gln-Thr (SEQ ID NO:54), Ala-Asp-Trp-Ser-Trp-Leu-Gln (SEQ ID NO:55), Ala-Asp-Trp-Ser-Trp-Leu-Gln-Thr (SEQ ID NO:56), Ala-Leu-Asp-Trp-Ser-Trp-Ala-Gln-Thr-Glu (SEQ ID NO:57), Leu-Asp-Trp-Ser-Trp-Ala-Gln-Thr-Glu (SEQ ID NO:58), Thr-Ala-Leu-Asp-Trp-Ser-Trp-Ala (SEQ ID NO:59), Ala-Leu-Asp-Trp-Ser-Trp-Ala-Gln-Thr-Glu (SEQ ID NO:60), Leu-Asp-Trp-Ser-Trp-Ala-Gln-Thr-Glu (SEQ ID NO:61), Leu-Asp-Trp-Ser-Trp-Ala (SEQ ID NO:62), Thr-Ala-Leu-Asp-Trp-Ser-Trp-Ala-Gln-Thr (SEQ ID NO:63), Thr-Ala-Leu-Asp-Trp-Ser-Trp-Ala-Gln (SEQ ID NO:64), Ala-Leu-Asp-Trp-Ser-Trp-Ala-Gln-Thr (SEQ ID NO:65), Leu-Asp-Trp-Ser-Trp-Ala-Gln (SEQ ID NO:66), Leu-Asp-Trp-Ser-Trp-Ala-Gln-Thr (SEQ ID NO:67), Thr-Ala-Ala-Asp-Trp-Ser-Trp-Ala-Gln-Thr-Glu (SEQ ID NO:68), Ala-Asp-Trp-Ser-Trp-Ala-Gln-Thr-Glu (SEQ ID NO:69), Thr-Ala-Ala-Asp-Trp-Ser-Trp-Ala (SEQ ID NO:70), Ala-Ala-Asp-Trp-Ser-Trp-Ala-Gln-Thr-Glu (SEQ ID NO:71), Ala-Asp-Trp-Ser-Trp-Ala-Gln-Thr-Glu (SEQ ID NO:72), Ala-Asp-Trp-Ser-Trp-Ala (SEQ ID NO:73), Thr-Ala-Ala-Asp-Trp-Ser-Trp-Ala-Gln-Thr (SEQ ID NO:74), Thr-Ala-Ala-Asp-Trp-Ser-Trp-Ala-Gln (SEQ ID NO:75), Ala-Ala-Asp-Trp-Ser-Trp-Ala-Gln-Thr (SEQ ID NO:76), Ala-Asp-Trp-Ser-Trp-Ala-Gln (SEQ ID NO:77), Ala-Asp-Trp-Ser-Trp-Ala-Gln-Thr (SEQ ID NO:78), Thr-Ala-Leu-Asp-Phe-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:79), Leu-Asp-Phe-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:80), Thr-Ala-Leu-Asp-Phe-Ser-Trp-Leu (SEQ ID NO:81), Ala-Leu-Asp-Phe-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:82), Leu-Asp-Phe-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:83), Leu-Asp-Phe-Ser-Trp-Leu (SEQ ID NO:84), Thr-Ala-Leu-Asp-Phe-Ser-Trp-Leu-Gln-Thr (SEQ ID NO:85), Thr-Ala-Leu-Asp-Phe-Ser-Trp-Leu-Gln (SEQ ID NO:86), Ala-Leu-Asp-Phe-Ser-Trp-Leu-Gln-Thr (SEQ ID NO:87), Leu-Asp-Phe-Ser-Trp-Leu-Gln (SEQ ID NO:88), Leu-Asp-Phe-Ser-Trp-Leu-Gln-Thr (SEQ ID NO:89), Thr-Ala-Leu-Asp-Tyr-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:90), Leu-Asp-Tyr-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:91), Thr-Ala-Leu-Asp-Tyr-Ser-Trp-Leu (SEQ ID NO:92), Ala-Leu-Asp-Tyr-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:93), Leu-Asp-Tyr-Ser-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:94), Leu-Asp-Tyr-Ser-Trp-Leu (SEQ ID NO:95), Thr-Ala-Leu-Asp-Tyr-Ser-Trp-Leu-Gln-Thr (SEQ ID NO:96), Thr-Ala-Leu-Asp-Tyr-Ser-Trp-Leu-Gln (SEQ ID NO:97), Ala-Leu-Asp-Tyr-Ser-Trp-Leu-Gln-Thr (SEQ ID NO:98), Leu-Asp-Tyr-Ser-Trp-Leu-Gln (SEQ ID NO:99), Leu-Asp-Tyr-Ser-Trp-Leu-Gln-Thr (SEQ ID NO:100), Thr-Ala-Leu-Asp-Trp-Ala-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:101), Leu-Asp-Trp-Ala-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:102), Thr-Ala-Leu-Asp-Trp-Ala-Trp-Leu (SEQ ID NO:103), Ala-Leu-Asp-Trp-Ala-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:104), Leu-Asp-Trp-Ala-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:105), Leu-Asp-Trp-Ala-Trp-Leu (SEQ ID NO:106), Thr-Ala-Leu-Asp-Trp-Ala-Trp-Leu-Gln-Thr (SEQ ID NO:107), Thr-Ala-Leu-Asp-Trp-Ala-Trp-Leu-Gln (SEQ ID NO:108), Ala-Leu-Asp-Trp-Ala-Trp-Leu-Gln-Thr (SEQ ID NO:109), Leu-Asp-Trp-Ala-Trp-Leu-Gln (SEQ ID NO:110), Leu-Asp-Trp-Ala-Trp-Leu-Gln-Thr (SEQ ID NO:111), Thr-Ala-Leu-Asp-Trp-Glu-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:112), Leu-Asp-Trp-Glu-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:113), Thr-Ala-Leu-Asp-Trp-Glu-Trp-Leu (SEQ ID NO:114), Ala-Leu-Asp-Trp-Glu-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:115), Leu-Asp-Trp-Glu-Trp-Leu-Gln-Thr-Glu (SEQ ID NO:116), Leu-Asp-Trp-Glu-Trp-Leu (SEQ ID NO:117), Thr-Ala-Leu-Asp-Trp-Glu-Trp-Leu-Gln-Thr (SEQ ID NO:118), Thr-Ala-Leu-Asp-Trp-Glu-Trp-Leu-Gln (SEQ ID NO:119), Ala-Leu-Asp-Trp-Glu-Trp-Leu-Gln-Thr (SEQ ID NO:120), Leu-Asp-Trp-Glu-Trp-Leu-Gln (SEQ ID NO:121), and Leu-Asp-Trp-Glu-Trp-Leu-Gln-Thr (SEQ ID NO:122).
7. The method of claim 1 , wherein X a consists of 6-12 amino acid residues.
8. The method of claim 1 , wherein X a consists of 6-10 amino acid residues.
9. The method of claim 1 , wherein X a comprises at least five basic amino acid residues.
10. The method of claim 1 , wherein X a comprises at least five amino acid residues independently selected from L-arginine, D-arginine, L-lysine and D-lysine.
11. The method of claim 1 , wherein X a is selected from the group consisting of Arg-Arg-Met-Lys-Trp-Lys-Lys (SEQ ID NO:123), Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg (SEQ ID NO:124), D-Tyr-D-Gly-D-Arg-D-Lys-D-Lys-D-Arg-D-Arg-D-Gln-D-Arg-D-Arg-D-Arg (SEQ ID NO:125), Tyr-Ala-Arg-Lys-Ala-Arg-Arg-Gln-Ala-Arg-Arg (SEQ ID NO:126), D-Tyr-D-Ala-D-Arg-D-Lys-D-Ala-D-Arg-D-Arg-D-Gln-D-Ala-D-Arg-D-Arg (SEQ ID NO:127), Tyr-Ala-Arg-Ala-Ala-Arg-Arg-Ala-Ala-Arg-Arg (SEQ ID NO:128), D-Tyr-D-Ala-D-Arg-D-Ala-D-Ala-D-Arg-D-Arg-D-Ala-D-Ala-D-Arg-D-Arg (SEQ ID NO:129), D-Arg-D-Arg-D-Met-D-Lys-D-Trp-D-Lys-D-Lys (SEQ ID NO:130), Arg-Arg-Arg-Arg-Arg-Arg (SEQ ID NO:149), Arg-Arg-Arg-Arg-Arg-Arg-Arg (SEQ ID NO:150), Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg (SEQ ID NO:151), Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg (SEQ ID NO:152), Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg (SEQ ID NO:153), Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg (SEQ ID NO:154), D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg (SEQ ID NO:155), D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg (SEQ ID NO:156), D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg (SEQ ID NO:157), D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg (SEQ ID NO:158), D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg (SEQ ID NO:159), and D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg (SEQ ID NO:160).
12. The method of claim 1 , wherein the compound is a small molecule.
13. The method of claim 1 , wherein the IKK is selected from the group consisting of IKKα and IKKβ.
14. The method of claim 1 , wherein the compound modulates NF-κB-dependent target gene expression.
15. The method of claim 14 , wherein the NF-κB-dependent target gene is E-selectin.
16. The method of claim 1 , wherein the compound inhibits osteoclast differentiation.
17. The method of claim 1 , wherein the cell is contacted in vivo.
18. The method of claim 1 , wherein the cell is contacted in vitro.
19. The method of claim 1 , wherein the anti-inflammatory compound inhibits NF-κB activation.
20. The method of claim 1 , wherein the anti-inflammatory compound modulates signal transduction involving IκB.
21. The method of claim 1 , wherein the membrane translocation domain is selected from the group consisting of the third helix of the antennapedia homeodomain and HIV-1 Tat protein.