# US 2013/0158093 A1 · GPR 17 Agonists and Screening Assay

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

- Number: US 2013/0158093 A1
- Document ID: US20130158093A1
- Status: Published application
- Kind: A1
- Application number: 13/775,433
- Filed: 2013-02-25
- Published: 2013-06-20
- Earliest claimed: 2011-09-07
- Assignees: Rheinische Friedrich-Wilhelms-Universität Bonn
- Inventors: Evi Kostenis; Andreas Spinrath; Stephanie Hennen; Lucas Peters; Christa E. Muller; Rhalid Akkari; Younis Baqi; Kirsten Ritter
- Web page: https://patentgrep.com/patent/US20130158093A1

## CPC

- C07D 209/42: Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
- A61K 31/405: Indole-alkanecarboxylic acids; Derivatives thereof, e.g. tryptophan, indomethacin
- A61P 9/00: Drugs for disorders of the cardiovascular system
- A61P 9/10: for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
- A61P 25/00: Drugs for disorders of the nervous system
- C07D 405/04: directly linked by a ring-member-to-ring-member bond
- C07D 409/04: directly linked by a ring-member-to-ring-member bond
- C12Q 1/025: {for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics (antimicrobial activity C12Q1/18)}
- G01N 33/5041: {involving analysis of members of signalling pathways}
- G01N 33/566: using specific carrier or receptor proteins as ligand binding reagents {where possible specific carrier or receptor proteins are classified with their target compounds}
- G01N 33/6896: {Neurological disorders, e.g. Alzheimer's disease}
- G01N 33/74: involving hormones {or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors}
- C07K 14/723: {G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH receptor}
- G01N 2333/4719: G-proteins
- G01N 2500/00: Screening for compounds of potential therapeutic value
- G01N 2800/285: Demyelinating diseases; Multipel sclerosis
- G01N 2800/2871: Cerebrovascular disorders, e.g. stroke, cerebral infarct, cerebral haemorrhage, transient ischemic event
- C07D: HETEROCYCLIC COMPOUNDS (macromolecular compounds C08)
- 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
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- 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)
- 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)

## Abstract

The present invention is related to a method of determining a test compound's ability to modify the biological activity of a GPR17. Said method comprises, among others, the step of contacting the test compound with a GPR17, or a functional GPR17 fragment in the presence of a suitable amount of a GPR17 agonist of formula I.

## Claims

### Claim at position 1

-7. (canceled)

### Claim 8

A method of treating, alleviating or preventing of a GPR17 mediated disease, optionally selected from ischemic diseases, stroke or multiple sclerosis, in a patient in need thereof comprising the steps of
(a) identifying a compound that modulates GPR17 biological activity by
(i) contacting a compound with a GPR17, or a functional GPR17 fragment in the presence of a suitable amount of a GPR17 agonist of formula I
or a salt thereof,
(ii) measuring the biological activity of said GPR17 or said functional GPR17 fragment after addition of said compound, and
(iii) comparing the biological activity measured in step (ii) with the biological activity of said GPR17 or said functional fragment thereof in the presence of said GPR 17 agonist of formula I without the addition of said compound to identify whether the compound is one that modulates GPR17 activity,
wherein in formula I
R1 and R2 are independently selected from the group comprising hydrogen, halogen, hydroxy, formyl, oxime, cyano, nitro, amino, NR6R7, carboxy, carbamoyl, (C1-C8)alkyl, (C1-C8) all yloxy, (C1-C8)alkylthio, (C2-C8)alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl, (C3-C8)cycloalkloxy, (C3-C8)cycloalkylamino, aryl, heteroaryl, aryloxy, heteroaryloxy, halogen, trifluoromethyl, (C1-C8)alkylcarbonyl, (C1-C8) alkylaminocarbonyl, di(C1-C8)alkylaminocarbonyl, arylcarbonyl, heteroarylcarbonyl, aryl(C1-C8)alkyl, heteroaryl(C1-C8)alkyl, aryl(C1-C8)alkyloxy, heteroaryl(C1-C8)alkyloxy, aryl(C1-C8)alkylcarbonyl, heteroaryl(C1-C8)alkylcarbonyl, aryl(C1-C8)alkyloxycarbonyl, heteroaryl(C1-C8)alkyloxycarbonyl, (C1-C8)alkyloxycarbonyl, (C1-C8)alkylsulfonyl, arylsulfonyl, heteroarylsulfonyl, sulfamoyl, sulfonylamino, (C1-C8)alkylaminosulfonyl, di(C1-C8)alkylaminosulfonyl, arylsulfonylamino, heteroarylsulfonylamino and (C1-C8)alkylsulfonylamino; wherein each alkyl, alkenyl alkynyl or cycloalkyl may be unsubstituted or substituted with one or more residues selected from among hydroxyl, (C1-C5)alkyloxy, (C1-C3)alkyloxy(C1-C3)alkyloxy, halogen, and NR6R7; and wherein each aryl or heteroaryl can be unsubstituted or substituted with one or more residues selected from among hydroxyl, (C1-C5)alkyloxy, halogen, (C1-C5)alkyl, (C3-C8)cycloalkyl, carboxy, NR6R7, cyano, trifluormethyl and nitro;
R3 is selected from hydrogen, a group —(CH2)mCH2—COOH, OH, NH, and (C1-C5)alkyl which is optionally substituted with one or more halogens, one or two hydroxyl groups or (C1-C3)alkoxy;
R4 is selected from hydrogen and fluoro;
R5 is selected from hydrogen, halogen, (C1-C3)alkyl, (C1-C3)alkyloxy, (C1-C3)alkylthio, (C2-C4)alkenvl, (C2-C4)alkynyl, and NR6R7;
R6 and R7 are independently selected from hydrogen, (C1-C6)alkyl, (C3-C7)cycloalkyl, phenyl, heteroaryl, phenyl(C1-C6)alkyl, heteroaryl(C1-C6)alkyl, (C1-C6)alkylsulfonyl, phenylsulfonyl, heteroarylsulfonyl, (C1-C6)alkylcarbonyl, (C1-C6)alkoxycarbonyl, aminocarbonyl, (C1-C6)alkylaminocarbonyl, phenylcarbonyl, and heteroarylcarbonyl; wherein each alkyl may be unsubstituted or substituted with one or more residues selected from among hydroxyl, (C1-C3)alkyloxy, phenyl, halo, carboxy, and NR8R9; and wherein R6 and R7 may form a 5- to 7-membered cycle; and wherein phenyl or heteroaryl can be unsubstituted or substituted with one or more residues selected from among hydroxyl, (C1-C3)alkyloxy, halogen, (C1-C3)alkyl, carboxy, NR8R9, cyano, trifluormethyl and nitro;
R8 and R9 are independently selected from among hydrogen and (C1-C3)alkyl;
n and m are independently 0, 1 or 2;
and
(b) administering said GPR17 modulating compound to the patient, optionally together with one or more pharmaceutically acceptable excipients.

### Claim 9 (depends on 8)

The method according to claim 8, wherein the GPR17 mediated disease is multiple sclerosis.

### Claim 10 (depends on 2)

A screening assay for the identification of GPR 17 modulators among a multitude of compounds, said screening assay comprising
(a) cells or membrane fractions expressing GPR17 or a functional GPR17 fragment and
(b) a suitable amount of a GPR17 agonist of formula I
wherein in formula I
R1 and R2 are independently selected from the group comprising hydrogen, halogen, hydroxy, formyl, oxime, cyano, nitro, amino, NR6R7, carboxy, carbamoyl, (C1-C8)alkyl, (C1-C8) alkyloxy, (C1-C8)alkylthio, (C2-C8)alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl, (C3-C8)cycloalkyloxy, (C3-C8)cycloalkylamino, aryl, heteroaryl, aryloxy, heteroaryloxy, halogen, trifluoromethyl, (C1-C8)alkylcarbonyl, (C1-C8) alkylaminocarbonyl, di(C1-C8)alkylaminocarbonyl, arylcarbonyl, heteroarylcarbonyl, aryl(C1-C8)alkyl, heteroaryl(C1-C8)alkyl, aryl(C1-C8)alkyloxy, heteroaryl(C1-C8)alkyloxy, aryl(C1-C8)alkylcarbonyl, heteroaryl(C1-C8)alkylcarbonyl, aryl(C1-C8)alkyloxycarbonyl, heteroaryl(C1-C8)alkyloxycarbonyl, (C1-C8)alkyloxycarbonyl, (C1-C8)alkylsulfonyl, arylsulfonyl, heteroarylsulfonyl, sulfamoyl, sulfonylamino, (C1-C8)alkylaminosulfonyl, di(C1-C8)alkylaminosulfonyl, arylsulfonylamino, heteroarylsulfonylamino and (C1-C8)alkylsulfonylamino; wherein each alkyl, alkenyl, alkynyl or cycloalkyl may be unsubstituted or substituted with one or more residues selected from among hydroxyl, (C1-C5)alkyloxy, (C1-C3)alkyloxy(C1-C3)alkyloxy, halogen, and NR6R7; and wherein each aryl or heteroaryl can be unsubstituted or substituted with one or more residues selected from among hydroxyl, (C1-C5)alkyloxy, halogen, (C1-C5)alkyl, (C3-C8)cycloalkyl, carboxy, NR6R7, cyano, trifluormethyl and nitro;
R3 is selected from hydrogen, a group —(CH2)mCH2—COOH, OH, NH, and (C1-C5)alkyl which is optionally substituted with one or more halogens, one or two hydroxyl groups or methoxy;
R4 is independently selected from hydrogen and fluoro;
R5 is selected from hydrogen, halogen, (C1-C3)alkyl, (C1-C3)alkyloxy, (C1-C3)alkylthio, (C2-C4)alkenyl, (C2-C4)alkynyl, and NR6R7;
R6 and R7 are independently selected from hydrogen,(C1-C6)alkyl, (C3-C7)cycloalkyl, phenyl, heteroaryl, phenyl(C1-C6)alkyl, heteroaryl(C1-C6)alkyl, (C1-C6)alkylsulfonyl, phenylsulfonyl, heteroarylsulfonyl,(C1-C6)alkylcarbonyl,(C1-C6)alkoxycarbonyl, aminocarbonyl, (C1-C6)alkylaminocarbonyl, phenylcarbonyl, and heteroarylcarbonyl; wherein each alkyl may be unsubstituted or substituted with one or more residues selected from among hydroxyl, (C1-C3)alkyloxy, phenyl, halo, carboxy, and NR8R9; and wherein R6 and R7 may form a 5- to 7-membered cycle; and wherein phenyl or heteroaryl can be unsubstituted or substituted with one or more residues selected from among hydroxyl, (C1-C3)alkyloxy, halogen, (C1-C3)alkyl, carboxy, NR8R9, cyano, trifluormethyl and nitro;
R8 and R9 are independently selected from among hydrogen and (C1-C3)alkyl;
n and m are independently 0, 1 or 2, or
a salt thereof,
and wherein, optionally, the GPR agonist of formula I is as defined in claim 2.

### Claim 11 (depends on 10)

The screening assay according to claim 10 further comprising means for determining the GPR17 activity,
wherein, optionally, said means for determining the GPR17 activity is selected from one or more of the group consisting of:
a. means for determining the release of calcium from intracellular calcium stores associated with GPR17 activation, said means optionally comprising a cell membrane permeable indicator which binds to calcium released within the cell thereby providing a measurable signal, preferably fluorescence or luminescence;
b. means for determining GTPYS binding to heterotrimeric G proteins, wherein said heterotrimeric G protein is optionally 35SGTPYS;
c. means for determining the inhibition of cAMP formation or its increase associated with GPR17 activation, said means optionally comprising a stimulator of the adenylyl cyclase and a suitable cAMP indicator system;
d. means for determining the increase in inositolphosphates (IP), quantified as IP1, said means optionally comprising a suitable IP 1 detector system; and
e. means for determining the recruitment of cytoplasmic β-arrestin proteins, optionally by detection of the B-arrestin translocation to the receptor using fluorescence or bioluminescence resonance energy transfer.

### Claim 12 (depends on 11)

The screening assay according to claim 11 (c) wherein said stimulator of the adenylyl cyclase is forskolin, and wherein said cAMP indicator system is a competitive immunoassay, optionally providing a fluorescence signal.

### Claim 13

(canceled)

### Claim 14

A compound selected from
3-(2-carboxyethyl)-4,6-dibromo-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-4,6-dichloro-(1-carboxyethyl)-indole-2-carboxylic acid,
3-(2-carboxyethyl)-6-bromo-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-6-iodo-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-4,6-diiodo-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-4,6-diphenyl-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-6-phenyl-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-6,7-dichloro-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-6-chloro-7-fluoro-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-6-bromo-2-carboxy-7-fluoro-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-6-benzyl-2-carboxy-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-4,6-dihydroxy-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl-6-(4-fluorophenyl)-1H-indol-2-carboxylic acid,
3-(2-carboxyethyl)-6-furanyl-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-6-thienyl-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-7-fluoro-6-phenyl-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-6-(4-fluorophenyl)-7-fluoro-1H-indol-2-carboxylic acid,
3 -(2-carboxyethyl)-6-furanyl-7-fluoro-1H-indole-2-carboxylic acid,
3 -(2-carboxyethyl)-6-thienyl-7-fluoro-1H-indole-2-carboxylic acid,
and salts of any of the foregoing.

### Claim 15 (depends on 8)

The method according to claim 8, wherein in formula I
R1 is selected from fluoro, chloro, bromo, iodo, (C1-C3)alkyl, (C1-C3)alkoxy, phenyl, phenyl(C1-C3)alkyl, phenyl(C1-C3)alkoxy, phenylcarbonyl, (C5-C6)heteroaryl, (C5-C6)heteroarylcarbonyl, (C5-C6)heteroaryl(C1-C3)alkyl and (C5-C6)heteroaryl(C1-C3)alkoxy, wherein the alkyl and alkoxy groups are optionally substituted with one or more halogens, (C1-C3)alkoxy, or hydroxyl, and wherein the phenyl and (C5-C6)heteroaryl groups can be substituted with one or more halogens, (C1-C3)alkoxy, (C1-C3)alkyl, NR6R7,or hydroxyl;
R2 is selected from hydrogen, fluoro, chloro, bromo, iodo, (C1-C3)alkyl, (C1-C3)alkoxy, and phenyl, wherein the alkyl and alkoxy groups are optionally substituted with one or more halogens, (C1-C3)alkoxy, or hydroxyl, and wherein the phenyl group can be substituted with one or more halogens, (C1-C3)alkoxy, (C1-C3)alkyl, or hydroxyl;
R3 is selected from hydrogen and a group —(CH2)mCH2—COOH;
R4 is hydrogen or fluoro, and is preferably hydrogen;
R5 is selected from hydrogen, halogen, methyl or methoxy, and preferably represents fluoro or hydrogen, and particularly hydrogen;
n and m are independently 0, 1 or 2.

### Claim 16 (depends on 8)

The method according to claim 8, wherein in formula I
R1 is selected from the group consisting of methyl, methoxy, hydroxy, CF3, chloro, fluoro, bromo, iodo, thienyl, furanyl, pyridyl, and phenyl which is optionally substituted with halogen, methyl or methoxy;
R2 is selected from the group consisting of hydrogen, methyl, methoxy, hydroxyl, CF3, chloro, fluoro, bromo, iodo and phenyl;
R3 is hydrogen, carboxymethyl, or carboxyethyl;
R4 and R5 are both hydrogen; and
n is 1.

### Claim 17 (depends on 8)

The method according to claim 8 wherein the GPR 17 agonist of formula I is selected from the group consisting of
3 -(2-carboxyethyl)-4,6-dichloro-1H-indole-2-carboxylic acid,
3 -(2-carboxyethyl)-4,6-dichloro-(1 -carboxyethyl)-indole-2-carboxylic acid,
3 -(2-carboxyethyl)-4,6-dimethyl-1H-indole-2-carboxylic acid,
3 -(2-carboxyethyl)-6-chloro-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-4,6-difluoro-1H-indole-2-carboxylic acid,
3 -(2-carboxyethyl)-4,6-dibromo-1H-indole-2-carboxylic acid,
3 -(2-carboxyethyl)-6-bromo-1H-indole-2-carboxylic acid,
3 -(2-carboxyethyl)-6-iodo-1H-indole-2-carboxylic acid,
3 -(2-carboxyethyl)-4,6-diiodo-1H-indole-2-carboxylic acid,
3 -(2-carboxyethyl)-4,6-dichloro-1H-indole-2-carboxylic acid,
3 -(2-carboxyethyl)-4,6-diphenyl-1H-indole-2-carboxylic acid,
3 -(2-carboxyethyl)-6-phenyl-1H-indole-2-carboxylic acid,
3 -(2-carboxyethyl)-6,7-dichloro-1H-indole-2-carboxylic acid,
3 -(2-carboxyethyl)-6-chloro-7-fluoro-1H-indole-2-carboxylic acid,
3 -(2-carboxyethyl)-6-bromo-2-carboxy-7-fluoro)-1H-indole-2-carboxylic acid,
3 -(2-carboxyethyl)-4,6-dimethoxy-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-6-phenoxy-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-6-benzyl-2-carboxy-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-4,6-dihydroxy-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-6-(4-fluorophenyl)-1H-indol-2-carboxylic acid,
3-(2-carboxyethyl)-6-furanyl-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-6-thienyl-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-7-fluoro-6-phenyl-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-6-(4-fluorophenyl)-7-fluoro-1H-indo1-2-carboxylic acid,
3-(2-carboxyethyl)-6-furanyl-7-fluoro-1H-indole-2-carboxylic acid,
3-(2-carboxyethyl)-6-thienyl-7-fluoro-1H-indole-2-carboxylic acid,
or a salt of any of the foregoing.

### Claim 18 (depends on 8)

The method according to claim 8, wherein the biological activity of GPR17 is determined by measuring one or more of the following parameters: 35SGTPYS binding, the inhibition of cAMP formation, the release of calcium from intracellular calcium stores, the increase in inositolphosphates (IP), and/or the recruitment of cytoplasmic β-arrestin proteins.

### Claim 19 (depends on 8)

The method according to claim 8, wherein the activity of GPR17 is measured in a transfected cell line or membrane fragments thereof, which is preferably selected from transfected CHO cells, astrocytoma cells, COST cells or HEK293 cells and/or in naturally expressing tissue or cells and membrane fragments thereof.

### Claim 20 (depends on 8)

The method according to claim 8 wherein step (a) comprises the additional steps of
(iv) selecting one or more analogs of the GPR 17 modulating compound(s) identified in steps (i) to (iii),
(v) subjecting the one or more GPR 17 modulating compound analogs of step (iv) to the method and/or assay of steps (i)-(iii) thereby determining the GPR 17 modulating properties of said GPR 17 modulating compound analogs, and
(vi) optionally repeating steps (iv)-(v) one or more times by producing further chemical analogs of the GPR 17 modulating compound(s) thereby identifying one or more GPR17 modulating compound(s) with improved GPR17 modulating properties.