Immunological reagents and uses therefor

Patent metadata

Field	Value
Publication number	US-10245262-B2
Application number	US-201515300023-A
Country	US
Kind code	B2
Filing date	Apr 1, 2015
Priority date	Apr 1, 2014
Publication date	Apr 2, 2019
Grant date	Apr 2, 2019

Abstract

The present invention provides ligands which bind to MR1, some of which induce MR1 to bind to MAIT cells thereby activating or inhibiting MAIT cell activation.

First claim

The invention claimed is:

1. An MR1-ligand subunit [MR1-L] which binds MAIT cells, wherein said ligand is represented by Formula (I):

or a salt, tautomer, or stereoisomer thereof
wherein:
R 1 is selected from the group consisting of:
X—C(O)—R 1 ′ (where R 1 ′ is H, or optionally substituted C 1 -C 6 alkyl, and X is independently a bond or a divalent linker selected from the group consisting of C 1 -C 3 optionally substituted alkylene, —NR 2 ′— optionally substituted C 1 -C 3 alkylene-, —O— optionally substituted C 1 -C 3 alkylene-, —S— optionally substituted C 1 -C 3 alkylene-, —S(O)— optionally substituted C 1 -C 3 alkylene-, —N═CR 2 ′—, —CR 2 ′═CR 2 ″—, —NR 2 ′—C(O)—, —O—C(O)—, or —S—C(O)— where each R 2 ′ and R 2 ″ is independently selected from H, halogen, CN, or optionally substituted C 1 -C 6 alkyl); —X′—C(O)NR 3 ′R 4 ′ (where R 3 ′ is H or optionally substituted C 1 -C 6 alkyl and R 4 ′ is optionally substituted C 1 -C 6 alkyl, OH, or CN or R 3 ′R 4 ′ together form an optionally substituted heterocyclyl or optionally substituted heteroaryl, and X′ is independently a bond or a C 1 -C 3 optionally substituted alkylene); —X″—C(O)OR 5 ′ (wherein R 5 ′ is H or optionally substituted C 1 -C 6 alkyl, and X″ is independently a bond or a C 1 -C 3 optionally substituted alkylene); —X′″—C(O)NHSO 2 R 6 ′ (wherein R 6 ′ is optionally substituted aryl, or optionally substituted C 1 -C 6 alkyl, and X′″ is independently a bond or a C 1 -C 3 optionally substituted alkylene); and —X″″—S(O) 2 NHR 7 ′ (wherein R 7 ′ is H, optionally substituted C 1 -C 6 alkyl, or optionally substituted aryl, and X″″ is independently a bond or a C 1 -C 3 optionally substituted alkylene);
R 2 is selected from the group consisting of optionally substituted C 1-8 alkyl, —NH(optionally substituted C 1-6 alkyl), —N(optionally substituted C 1-6 alkyl)(optionally substituted aryl), —N(optionally substituted C 1-6 alkyl) 2 , —O(optionally substituted C 1-6 alkyl), —OC(O)(C 1-6 alkyl), —S(optionally substituted C 1-6 alkyl), —SC(O)(C 1-6 alkyl), and —S(O)(optionally substituted C 1-6 alkyl); and
Y and Z are oxo.

2. The [MR1-L] of claim 1 , wherein
R 1 is —X—C(O)—R 1 ′ (where R 1 ′ is H, or optionally substituted C 1 -C 6 alkyl, and X is independently a divalent linker selected from the group consisting of C 1 -C 3 optionally substituted alkylene, —NR 2 ′— optionally substituted C 1 -C 3 alkylene-, —O— optionally substituted C 1 -C 3 alkylene-, —N═CR 2 ′—, —CR 2 ′═CR 2 ″—, —NR 2 ′—C(O)—, —OC(O)—, or —SC(O)— where each R 2 ′ and R 2 ″ is independently selected from H or optionally substituted C 1 -C 6 alkyl); and
R 2 is selected from the group consisting of optionally substituted C 1-8 alkyl, NH(optionally substituted C 1-6 alkyl), —N(optionally substituted C 1-6 allyl)(optionally substituted aryl), —N(optionally substituted C 1-6 alkyl) 2 , —O(optionally substituted C 1-6 alkyl), —OC(O)(C 1-6 alkyl), —S(optionally substituted C 1-6 alkyl), —SC(O)(C 1-6 alkyl), or —S(O)(optionally substituted C 1-6 alkyl).

3. The [MR1-L] of claim 1 , wherein
R 1 is —X—C(O)R 1 ′ where R 1 ′ is H or C 1-6 alkyl and X is independently —NR 2 ′—CH 2 —, —N═CR 2 ′—, —CR 2 ′═CR 2 ″—, —NR 2 ′—C(O)—, —OC(O)—, or —SC(O)— where R 2 ′ and R 2 ″ are independently H or C 1-6 alkyl; and
R 2 is —NH—C 2 -C 6 alkyl optionally substituted 1 to 6 times with OH, OR 1 ′, NH 2 , NHR 1 ′, NR 1 ′R 2 ′, SH, or SR 1 ′, where R 1 ′ and R 2 ′ are independently C 1-6 alkyl, C 1-6 acyl, C 1-6 amido, or C 1-6 thioamido.

4. The [MR1-L] of claim 1 , wherein
R 1 is —N═CR 2 ′—C(O)R 1 ′ or —CH═CR 2 ″—C(O)R 1 ′, where each R 1 ′ and R 2 ′ is independently H or C 1-6 alkyl; and
R 2 is —NH—C 2 -C 6 alkyl optionally substituted 1 to 4 times with OH.

5. The [MR1-L] of claim 1 , wherein R 1 ′ is H or C 1 -C 6 alkyl optionally substituted by a group selected from halogen, hydroxy, mercapto or amino.

6. The [MR1-L] of claim 1 , wherein said ligand is selected from the group consisting of:

or a salt, tautomer, or stereoisomer thereof.

7. The [MR1-L] of claim 1 , wherein the optional substituents comprise a substituent selected from the group consisting of halogen, oxo, thio, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, C 2 -C 6 alkenyl, halo C 1 -C 6 alkyl, halo C 1 -C 6 alkoxy, —OH, phenyl, benzyl, phenoxy, benzyloxy, —NH 2 , —NHC 1 -C 4 alkyl, —N(C 1 -C 4 alkyl) 2 , —NHC 1 -C 4 acyl, —NHC(O)NH 2 , —NHC(O)NHC 1 -C 4 alkyl, —NHC(O)N(C 1 -C 4 alkyl) 2 , —NHC(S)NH 2 , —NHC(S)C 1 -C 4 alkyl, —NHC(S)N(C 1 -C 4 alkyl) 2 , guanidino, —CN, —NO 2 , mercapto, —S(O 2 )NH 2 , —S(O 2 )NHC 1 -C 4 alkyl, CO 2 H, CO 2 NH 2 and CO 2 NHC 1 -C 4 alkyl.

8. The [MR1-L] of claim 1 , wherein said ligand is selected from the group consisting of 5-(2-oxoethylideneamino)-6- D -ribitylaminouracil (5-OE-RU), 5-(2-oxopropylideneamino)-6- D -ribitylaminouracil (5-OP-RU), which were either isolated or generated in situ by mixing 5-amino-6- D -ribitylaminouracil (5-A-RU) with glyoxal or methylglyoxal, respectively, or functional analogs thereof, to oxidised forms thereof and reduced forms thereof.

9. The [MR1-L] of claim 1 which is in the form of a multimeric complex, wherein said multimeric complex comprises the formula [MR1-L] n , wherein n≥2 and ≤50.

10. The multimeric complex of claim 9 , wherein n=4.

11. The [MR1-L] of claim 1 , wherein said binding modulates MAIT cells.

12. The [MR1-L] of claim 11 , wherein said modulation is MAIT cell activation.

13. The [MR1-L] of claim 1 , wherein the MR1 comprises all or part of SEQ ID NO: 1, SEQ ID NO: 4, or a functional derivative thereof having one or more amino acid substitutions, additions and/or deletions to SEQ ID NO: 1 or SEQ ID NO: 4.

14. The [MR1-L] of claim 13 , wherein said MR1 comprises at least one mutation selected from the list consisting of K43A, K43M, K43I, K43L, K43F, K43Q, Y7A, Y7W, R9K, R9A, S24F, Y62A, L66A, L66F, W69A, R94K, R94A, I96A, I96F and W156A.

15. The [MR1-L] of claim 14 , wherein said MR1 comprises one or more mutations in surface exposed groups selected from the list consisting of D57, R61, L65, M72, V75, R79, T138, Q141, N146, H148, L151, N155, E158, and R167.

16. A compound selected from the group consisting of:

or a salt, tautomer, or stereoisomer thereof.

17. The compound according to claim 16 , wherein said compound is selected from the group consisting of 5-(2-oxoethylideneamino)-6- D -ribitylaminouracil (5-OE-RU), 5-(2-oxopropylideneamino)-6- D -ribitylaminouracil (5-OP-RU), which were either isolated or generated in situ by mixing 5-amino-6- D -ribitylaminouracil (5-A-RU) with glyoxal or methylglyoxal, respectively, or functional analogs thereof, oxidised forms thereof and reduced forms thereof.

18. A method of detecting the presence of MAIT cells in a biological sample from a subject, the method comprising the steps of a) contacting the biological sample with antigen presenting cells expressing MR1 bound to ligand,

Assignees

• Univ Queensland
• Univ Monash
• Univ Melbourne

Inventors

• Corbett Alexandra Jane
• Mccluskey James
• Kjer-Nielsen Lars
• Chen Zhenjun
• Rossjohn Jamie
• Patel Onisha
• Birkinshaw Richard William
• Eckle Sidonia Barbara Guiomar
• Fairlie David Paul
• Liu Ligong
• Mak Jeffrey Yam Wing

Classifications

• A61P37/00

  Drugs for immunological or allergic disorders · CPC title

• C07K14/70539

  MHC-molecules, e.g. HLA-molecules · CPC title

• C07D239/60

  Three or more oxygen or sulfur atoms · CPC title

• C07D239/545

  with other hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms · CPC title

• C07D239/54

  as doubly bound oxygen atoms or as unsubstituted hydroxy radicals · CPC title