Inhibitors of human EZH2, and methods of use thereof

Patent metadata

Field	Value
Publication number	US-9334527-B2
Application number	US-201414290642-A
Country	US
Kind code	B2
Filing date	May 29, 2014
Priority date	Sep 10, 2010
Publication date	May 10, 2016
Grant date	May 10, 2016

Abstract

The invention relates to inhibition of wild-type and certain mutant forms of human histone methyltransferase EZH2, the catalytic subunit of the PRC2 complex which catalyzes the mono- through tri-methylation of lysine 27 on histone H3 (H3-K27). In one embodiment the inhibition is selective for the mutant form of the EZH2, such that trimethylation of H3-K27, which is associated with certain cancers, is inhibited. The methods can be used to treat cancers including follicular lymphoma and diffuse large B-cell lymphoma (DLBCL). Also provided are methods for identifying small molecule selective inhibitors of the mutant forms of EZH2 and also methods for determining responsiveness to an EZH2 inhibitor in a subject.

First claim

We claim:

1. A method of treating cancer in a subject in need thereof comprising
detecting the presence of a mutation in the EZH2 substrate pocket domain of SEQ ID NO: 7, in a sample from the subject, wherein said detecting comprises screening, at amino acid positions Y641, A677, and A687 of EZH2 of SEQ ID NO: 1, for the presence of at least one mutation, wherein the mutation at amino acid position Y641 is selected from the group consisting of a substitution of phenylalanine (F) for the wild type residue tyrosine (Y) at amino acid position 641 of SEQ ID NO: 1 (Y641F); a substitution of histidine (H) for the wild type residue tyrosine (Y) at amino acid position 641 of SEQ ID NO: 1 (Y641H); a substitution of asparagine (N) for the wild type residue tyrosine (Y) at amino acid position 641 of SEQ ID NO: 1 (Y641N); a substitution of serine (S) for the wild type residue tyrosine (Y) at amino acid position 641 of SEQ ID NO: 1 (Y641S); and a substitution of cysteine (C) for the wild type residue tyrosine (Y) at amino acid position 641 of SEQ ID NO: 1 (Y641C), wherein the mutation at amino acid position A677 is a substitution of glycine (G) for the wild type residue alanine (A) at amino acid position 677 of SEQ ID NO: 1 (A677G), wherein the mutation at amino acid position A687 is a substitution of valine (V) for the wild type residue alanine (A) at amino acid position 687 of SEQ ID NO: 1 (A687V), and wherein the mutation increases EZH2 trimethylation of Lys27 of histone H3 (H3-K27); and
identifying the subject as a candidate for treatment with an EZH2 inhibitor based on the presence of a detected EZH2 mutation;
administering a therapeutically effective amount of an EZH2 inhibitor to the identified subject, wherein the EZH2 inhibitor inhibits the conversion of H3-K27 to trimethylated H3-K27.

2. The method of claim 1 , wherein the nucleic acid encoding SEQ ID NO: 7 is SEQ ID NO: 8.

3. The method of claim 1 , wherein the EZH2 inhibitor is a small molecule.

4. The method of claim 1 , wherein the cancer is lymphoma, selected from the group consisting of non-Hodgkin lymphoma, follicular lymphoma, diffuse large B-cell lymphoma and diffuse large B-cell lymphoma of germinal center B cell-like subtype.

5. The method of claim 1 , wherein the EZH2 inhibitor is selected from the group consisting of

or a pharmaceutically acceptable salt thereof.

6. The method of claim 1 wherein the EZH2 inhibitor is

or a pharmaceutically acceptable salt thereof.

7. The method of claim 6 , wherein the cancer is lymphoma.

8. The method of claim 7 , wherein the lymphoma is non-Hodgkin lymphoma.

9. The method of claim 7 , wherein the lymphoma is follicular lymphoma.

10. The method of claim 7 , wherein the lymphoma is diffuse large B-cell lymphoma.

11. The method of claim 7 , wherein the lymphoma is diffuse large B-cell lymphoma of germinal center B cell-like subtype.

12. The method of claim 2 , wherein the EZH2 inhibitor) is

or a pharmaceutically acceptable salt thereof.

13. The method of claim 4 , wherein the EZH2 inhibitor is

or a pharmaceutically acceptable salt thereof.

Assignees

• Epizyme Inc

Inventors

• Kuntz Kevin Wayne
• Knutson Sarah Kathleen
• Wigle Timothy James Nelson

Classifications

• A61P35/02

  specific for leukemia · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• G01N33/57505

  of the blood, e.g. leukaemia · CPC title

• C12Q1/68 Primary

  involving nucleic acids · CPC title

• C07D473/34

  attached in position 6, e.g. adenine · CPC title