# US 10,245,262 B2 · Immunological reagents and uses therefor

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,245,262 B2
- Document ID: US10245262B2
- Status: Granted
- Kind: B2
- Application number: 15/300,023
- Filed: 2015-04-01
- Published: 2017-07-27
- Granted: 2019-04-02
- Earliest claimed: 2014-04-01
- Assignees: The University of Queensland; Monash University; The University of Melbourne
- Applicants: The University of Queensland; Monash University; The University of Melbourne
- Inventors: Alexandra Jane Corbett; James McCluskey; Lars Kjer-Nielsen; Zhenjun Chen; Jamie Rossjohn; Onisha Patel; Richard William Birkinshaw; Sidonia Barbara Guiomar Eckle; David Paul Fairlie; Ligong Liu; Jeffrey Yam Wing Mak
- Web page: https://patentgrep.com/patent/US10245262B2

## CPC

- A61K 31/513: having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
- A61K 47/55: the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug
- A61K 47/6425: {the peptide or protein in the drug conjugate being a receptor, e.g. CD4, a cell surface antigen, i.e. not a peptide ligand targeting the antigen, or a cell surface determinant, i.e. a part of the surface of a cell}
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P 35/00: Antineoplastic agents
- A61P 37/00: Drugs for immunological or allergic disorders
- C07D 239/54: as doubly bound oxygen atoms or as unsubstituted hydroxy radicals
- C07D 239/545: 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
- C07D 239/60: Three or more oxygen or sulfur atoms
- C07K 14/70539: {MHC-molecules, e.g. HLA-molecules}
- G01N 33/56972: {White blood cells}
- A61K 38/00: 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)
- G01N 2333/70539: MHC-molecules, e.g. HLA-molecules
- G01N 2800/26: Infectious diseases, e.g. generalised sepsis
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C07D: HETEROCYCLIC COMPOUNDS (macromolecular compounds C08)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## 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.

## Claims

### Claim 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:
R1 is selected from the group consisting of:
X—C(O)—R1′ (where R1′ is H, or optionally substituted C1-C6alkyl, and X is independently a bond or a divalent linker selected from the group consisting of C1-C3 optionally substituted alkylene, —NR2′— optionally substituted C1-C3alkylene-, —O— optionally substituted C1-C3alkylene-, —S— optionally substituted C1-C3alkylene-, —S(O)— optionally substituted C1-C3alkylene-, —N═CR2′—, —CR2′═CR2″—, —NR2′—C(O)—, —O—C(O)—, or —S—C(O)— where each R2′ and R2″ is independently selected from H, halogen, CN, or optionally substituted C1-C6alkyl); —X′—C(O)NR3′R4′ (where R3′ is H or optionally substituted C1-C6alkyl and R4′ is optionally substituted C1-C6alkyl, OH, or CN or R3′R4′ together form an optionally substituted heterocyclyl or optionally substituted heteroaryl, and X′ is independently a bond or a C1-C3 optionally substituted alkylene); —X″—C(O)OR5′ (wherein R5′ is H or optionally substituted C1-C6alkyl, and X″ is independently a bond or a C1-C3 optionally substituted alkylene); —X′″—C(O)NHSO2R6′ (wherein R6′ is optionally substituted aryl, or optionally substituted C1-C6alkyl, and X′″ is independently a bond or a C1-C3 optionally substituted alkylene); and —X″″—S(O)2NHR7′ (wherein R7′ is H, optionally substituted C1-C6alkyl, or optionally substituted aryl, and X″″ is independently a bond or a C1-C3 optionally substituted alkylene);
R2 is selected from the group consisting of optionally substituted C1-8alkyl, —NH(optionally substituted C1-6alkyl), —N(optionally substituted C1-6alkyl)(optionally substituted aryl), —N(optionally substituted C1-6alkyl)2, —O(optionally substituted C1-6alkyl), —OC(O)(C1-6alkyl), —S(optionally substituted C1-6alkyl), —SC(O)(C1-6alkyl), and —S(O)(optionally substituted C1-6alkyl); and
Y and Z are oxo.

### Claim 2 (depends on 1)

The [MR1-L] of claim 1, wherein
R1 is —X—C(O)—R1′ (where R1′ is H, or optionally substituted C1-C6alkyl, and X is independently a divalent linker selected from the group consisting of C1-C3 optionally substituted alkylene, —NR2′— optionally substituted C1-C3alkylene-, —O— optionally substituted C1-C3alkylene-, —N═CR2′—, —CR2′═CR2″—, —NR2′—C(O)—, —OC(O)—, or —SC(O)— where each R2′ and R2″ is independently selected from H or optionally substituted C1-C6alkyl); and
R2 is selected from the group consisting of optionally substituted C1-8alkyl, NH(optionally substituted C1-6alkyl), —N(optionally substituted C1-6allyl)(optionally substituted aryl), —N(optionally substituted C1-6alkyl)2, —O(optionally substituted C1-6alkyl), —OC(O)(C1-6alkyl), —S(optionally substituted C1-6alkyl), —SC(O)(C1-6alkyl), or —S(O)(optionally substituted C1-6alkyl).

### Claim 3 (depends on 1)

The [MR1-L] of claim 1, wherein
R1 is —X—C(O)R1′ where R1′ is H or C1-6alkyl and X is independently —NR2′—CH2—, —N═CR2′—, —CR2′═CR2″—, —NR2′—C(O)—, —OC(O)—, or —SC(O)— where R2′ and R2″ are independently H or C1-6alkyl; and
R2 is —NH—C2-C6 alkyl optionally substituted 1 to 6 times with OH, OR1′, NH2, NHR1′, NR1′R2′, SH, or SR1′, where R1′ and R2′ are independently C1-6alkyl, C1-6acyl, C1-6amido, or C1-6thioamido.

### Claim 4 (depends on 1)

The [MR1-L] of claim 1, wherein
R1 is —N═CR2′—C(O)R1′ or —CH═CR2″—C(O)R1′, where each R1′ and R2′ is independently H or C1-6alkyl; and
R2 is —NH—C2-C6 alkyl optionally substituted 1 to 4 times with OH.

### Claim 5 (depends on 1)

The [MR1-L] of claim 1, wherein R1′ is H or C1-C6alkyl optionally substituted by a group selected from halogen, hydroxy, mercapto or amino.

### Claim 6 (depends on 1)

The [MR1-L] of claim 1, wherein said ligand is selected from the group consisting of:
or a salt, tautomer, or stereoisomer thereof.

### Claim 7 (depends on 1)

The [MR1-L] of claim 1, wherein the optional substituents comprise a substituent selected from the group consisting of halogen, oxo, thio, C1-C6alkyl, C1-C6alkoxy, C2-C6alkenyl, halo C1-C6alkyl, halo C1-C6alkoxy, —OH, phenyl, benzyl, phenoxy, benzyloxy, —NH2, —NHC1-C4alkyl, —N(C1-C4alkyl)2, —NHC1-C4acyl, —NHC(O)NH2, —NHC(O)NHC1-C4alkyl, —NHC(O)N(C1-C4alkyl)2, —NHC(S)NH2, —NHC(S)C1-C4alkyl, —NHC(S)N(C1-C4alkyl)2, guanidino, —CN, —NO2, mercapto, —S(O2)NH2, —S(O2)NHC1-C4alkyl, CO2H, CO2NH2 and CO2NHC1-C4alkyl.

### Claim 8 (depends on 1)

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.

### Claim 9 (depends on 1)

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.

### Claim 10 (depends on 9)

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

### Claim 11 (depends on 1)

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

### Claim 12 (depends on 11)

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

### Claim 13 (depends on 1)

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.

### Claim 14 (depends on 13)

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.

### Claim 15 (depends on 14)

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.

### Claim 16

A compound selected from the group consisting of:
or a salt, tautomer, or stereoisomer thereof.

### Claim 17 (depends on 16)

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.

### Claim 18 (depends on 16)

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, under conditions that would allow binding of the MR1 with MAIT cells present in the sample; and b) detecting the presence of MAIT cell activity, wherein said ligand is defined in claim 16 or is a compound according to claim 16.

### Claim 19 (depends on 1)

A pharmaceutical composition comprising an MR1 ligand subunit of claim 1 together with one or more pharmaceutically acceptable carriers and/or diluents.

### Claim 20 (depends on 16)

A pharmaceutical composition comprising a compound of claim 16 together with one or more pharmaceutically acceptable carriers and/or diluents.