NZ725914B2 - Inhibitors of lysine specific demethylase-1 - Google Patents

Inhibitors of lysine specific demethylase-1 Download PDF

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Publication number
NZ725914B2
NZ725914B2 NZ725914A NZ72591415A NZ725914B2 NZ 725914 B2 NZ725914 B2 NZ 725914B2 NZ 725914 A NZ725914 A NZ 725914A NZ 72591415 A NZ72591415 A NZ 72591415A NZ 725914 B2 NZ725914 B2 NZ 725914B2
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Prior art keywords
compound
optionally substituted
benzonitrile
acceptable salt
pharmaceutically acceptable
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NZ725914A
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NZ725914A (en
Inventor
Young K Chen
Stephen W Kaldor
Toufike Kanouni
Jefrey Alan Stafford
James Marvin Veal
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Celgene Quanticel Research Inc
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Publication date
Application filed by Celgene Quanticel Research Inc filed Critical Celgene Quanticel Research Inc
Priority claimed from PCT/US2015/028635 external-priority patent/WO2015168466A1/en
Publication of NZ725914A publication Critical patent/NZ725914A/en
Publication of NZ725914B2 publication Critical patent/NZ725914B2/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/4545—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring hetero atom, e.g. pipamperone, anabasine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/513—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A61K31/551—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole having two nitrogen atoms, e.g. dilazep
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/05—Isotopically modified compounds, e.g. labelled
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B59/00—Introduction of isotopes of elements into organic compounds ; Labelled organic compounds per se
    • C07B59/002—Heterocyclic compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with 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
    • C07D239/32—One oxygen, sulfur or nitrogen atom
    • C07D239/34—One oxygen atom
    • C07D239/36—One oxygen atom as doubly bound oxygen atom or as unsubstituted hydroxy radical
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with 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
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/47—One nitrogen atom and one oxygen or sulfur atom, e.g. cytosine
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing three or more hetero rings
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/10—Spiro-condensed systems

Abstract

The present invention relates generally to compositions and methods for treating cancer and neoplastic disease. Provided herein are substituted heterocyclic derivative compounds and pharmaceutical compositions comprising said compounds. The subject compounds and compositions are useful for inhibition of lysine specific demethylase-1. Furthermore, the subject compounds and compositions are useful for the treatment of cancer.

Description

TORS OF LYSINE SPECIFIC DEMETHYLASE-l CROSS REFERENCE This application claims the bene?t ofUS. Provisional Application No. 61/987,354, ?led May 1, 2014, the contents ofwhich are hereby incorporated by reference in their entireties.
BACKGROUND A need exists in the art for an effective treatment of cancer and neoplastic disease.
BRIEF SUMMARY OF THE INVENTION ed herein are substituted heterocyclic derivative nds and pharmaceutical compositions comprising said compounds. The subject compounds and compositions are useful for inhibition lysine specific demethylase-l (LSD-l).
Furthermore, the subject compounds and itions are useful for the ent of cancer, such as acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), neuroblastoma, small round blue cell tumors, glioblastoma, prostate cancer, breast cancer, r cancer, lung cancer and/or melanoma and the like. The substituted heterocyclic derivative compounds described herein are based upon a central heterocyclic ring system, such as pyrimidinone, or the like. Said pyrimidinone ring system is ?arther substituted with a 4-cyanophenyl group and additional groups, such as aryl, heteroaryl or heterocyclic groups.
One embodiment provides a compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof, wherein, W is N, C-H, or C-F; X is hydrogen, halogen, -CN, optionally substituted alkyl, optionally substituted alkynyl, optionally substituted carbocyclylalkynyl, optionally substituted aryl, or ally substituted heteroaryl; Y is hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, or optionally substituted cycloalkylalkyl; Z is an optionally substituted group chosen from alkyl, carbocyclyl, C-attached cyclyl, N-attached heterocyclyl, heterocyclylalkyl, heterocyclylalkenyl, -O- heterocyclyl, -N(R)-heterocyclyl, -O-heterocyclylalkyl, -N(R)-heterocyclylalkyl, - N(R)(C1-C4alkylene)-NR2, -O(C1-C4alkylene)-NR2, and R is hydrogen or C1-C4alkyl.
One embodiment es a compound having the structure of Formula (I a), or a pharmaceutically able salt thereof, wherein, W is N, C-H, or C-F; X is hydrogen, n, -CN, optionally substituted alkynyl, optionally tuted carbocyclylalkynyl, optionally substituted aryl, or optionally substituted heteroaryl; Y is hydrogen, optionally substituted alkyl, optionally substituted lkyl, or optionally substituted cycloalkylalkyl; and Z is an optionally substituted group chosen from N—attached heterocyclyl, -O- heterocyclylalkyl, -N(H)-heterocyclyl, -N(Me)-heterocyclyl, -N(H)-heterocyclylalkyl, or -N(Me)-heterocyclylalkyl.
One embodiment provides a compound having the structure of Formula (lb), or a ceutically acceptable salt thereof, wherein, W is N, C-H, or C-F; X is hydrogen, n, optionally substituted l, optionally substituted carbocyclylalkynyl, optionally substituted aryl, or optionally substituted heteroaryl; Y is hydrogen, optionally substituted alkyl, or optionally substituted cycloalkyl; Z is an optionally substituted group chosen from N-heterocyclyl, -O- heterocyclylalkyl, -N(H)-heterocyclylalkyl, or -N(Me)-heterocyclylalkyl.
One embodiment provides a pharmaceutical composition comprising a compound ofFormula (I), or a pharmaceutically able salt thereof, and a pharmaceutically acceptable excipient. One ment provides a pharmaceutical composition comprising a compound of Formula (I a), or a ceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. One ment provides a pharmaceutical ition comprising a compound of Formula (lb), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.
One embodiment provides a method ofregulating gene transcription in a cell comprising inhibiting lysine-specific demethylase 1 activity by exposing the lysine- specif1c demethylase l enzyme to a compound of Formula (1). One embodiment provides a method ofregulating gene transcription in a cell comprising inhibiting lysine-specific demethylase 1 ty by exposing the lysine-specific demethylase l enzyme to a compound ofFormula (Ia). One embodiment provides a method ofregulating gene transcription in a cell comprising inhibiting lysine-specific demethylase 1 activity by exposing the -specific demethylase l enzyme to a compound of Formula (Ib).
One embodiment provides a method oftreating cancer in a patient in need thereof, comprising administering to the patient a compound of Formula (I), or a pharmaceutically acceptable salt thereof One embodiment provides a method of ng cancer in a t in need thereof, comprising administering to the patient a nd of Formula (Ia), or a pharmaceutically acceptable salt thereof One embodiment provides a method oftreating cancer in a patient in need thereof, comprising administering to the patient a compound of Formula (lb), or a pharmaceutically acceptable salt thereof INCORPORATION BY NCE All ations, patents, and patent applications mentioned in this specification are herein incorporated by reference for the c purposes identi?ed herein.
DETAILED DESCRIPTION OF THE ION As used herein and in the appended claims, the singular forms "a," "and," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "an agen " includes a plurality of such agents, and reference to "the cell" includes reference to one or more cells (or to a plurality of cells) and equivalents thereofknown to those skilled in the art, and so forth. When ranges are used herein for physical properties, such as molecular weight, or chemical properties, such as chemical ae, all combinations and subcombinations ofranges and speci?c ments therein are intended to be ed. The term "about" when referring to a number or a numerical range means that the number or numerical range referred to is an approximation within experimental variability (or within statistical experimental error), and thus the number or numerical range, in some instances, will vary between 1% and % ofthe stated number or numerical range. The term "comprising" (and related terms such as "comprise" or "comprises" or "having" or "including") is not intended to exclude that in other certain embodiments, for example, an embodiment of any composition of matter, composition, method, or process, or the like, described herein, st of‘ or "consist essentially of’ the described features.
De?nitions As used in the speci?cation and appended , unless ed to the contrary, the ing terms have the meaning indicated below.
"Amino" refers to the —NH2 radical.
"Cyano" refers to the -CN radical. " refers to the -N02 radical.
"Oxa" refers to the -O- l.
"Oxo" refers to the =0 radical. o" refers to the =S radical.
"Imino" refers to the =N-H radical.
"Oximo" refers to the =N-OH radical.
"Hydrazino" refers to the =N-NH2 l.
"Alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to ?fteen carbon atoms (e.g., C1-C15 alkyl). In certain embodiments, an alkyl comprises one to thirteen carbon atoms (e.g., C1-C13 alkyl). In certain embodiments, an alkyl comprises one to eight carbon atoms (e.g., C1-C8 alkyl). In other embodiments, an alkyl comprises one to ?ve carbon atoms (e.g., C1-C5 alkyl). In other embodiments, an alkyl comprises one to four carbon atoms (e.g., C1-C4 alkyl). In other ments, an alkyl comprises one to three carbon atoms (e.g., C1-C3 alkyl). In other embodiments, an alkyl comprises one to two carbon atoms (e.g., C1-C2 . In other embodiments, an alkyl comprises one carbon atom (e.g., C1 . In other embodiments, an alkyl comprises ?ve to ?fteen carbon atoms (e.g., C5-C15 alkyl). In other embodiments, an alkyl comprises ?ve to eight carbon atoms (e.g., C5-C8 alkyl). In other embodiments, an alkyl comprises two to ?ve carbon atoms (e.g., C2-C5 alkyl). In other embodiments, an alkyl comprises three to ?ve carbon atoms (e.g., C3-C5 alkyl). In other embodiments, the alkyl group is selected from methyl, ethyl, l-propyl (n-propyl), l-methylethyl (iso-propyl), l- butyl (n-butyl), l-methylpropyl (sec-butyl), 2-methylpropyl (iso-butyl), l,l-dimethylethyl butyl), yl (n-pentyl). The alkyl is attached to the rest of the molecule by a single bond. Unless stated otherwise speci?cally in the speci?cation, an alkyl group is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, -ORa, - SRa, -OC(O)-Ra, -N(Ra)2, -C(O)Ra, -C(O)ORa, -C(O)N(Ra)2, -N(Ra)C(O)ORa, -OC(O)— N (Ra)2, -N(Ra)C(O)Ra, -N(Ra)S(O)tRa (where t is l or 2), -S(O)tORa (where t is l or 2), -S(O)tRa (where t is l or 2) and -S(O)tN(Ra)2 (where t is l or 2) where each R21 is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, y, or tri?uoromethyl), ?uoroalkyl, carbocyclyl (optionally tuted with n, hydroxy, methoxy, or tri?uoromethyl), carbocyclylalkyl (optionally substituted with n, hydroxy, methoxy, or tri?uoromethyl), aryl nally substituted with halogen, hydroxy, methoxy, or romethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), heterocyclyl (optionally tuted with halogen, hydroxy, methoxy, or tri?uoromethyl), heterocyclylalkyl (optionally substituted with n, hydroxy, methoxy, or tri?uoromethyl), heteroaryl nally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), or heteroarylalkyl (optionally tuted with halogen, hydroxy, methoxy, or tri?uoromethyl).
"Alkoxy" refers to a radical bonded through an oxygen atom ofthe formula — O-alkyl, where alkyl is an alkyl chain as de?ned above.
"Alkenyl" refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, ning at least one carbon-carbon double bond, and having from two to twelve carbon atoms. In certain embodiments, an alkenyl comprises two to eight carbon atoms. In other embodiments, an alkenyl comprises two to four carbon atoms. The alkenyl is attached to the rest of the molecule by a single bond, for example, ethenyl (z'.e., vinyl), prop-l-enyl (z'.e., allyl), but-l-enyl, pent-l-enyl, penta-l,4-dienyl, and the like. Unless stated otherwise speci?cally in the speci?cation, an alkenyl group is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, -ORa, - SRa, -OC(O)-Ra, 2, -C(O)Ra, -C(O)ORa, -C(O)N(Ra)2, -N(Ra)C(O)ORa, -OC(O)— N (Ra)2, -N(Ra)C(O)Ra, -N(Ra)S(O)tRa (where t is l or 2), -S(O)tORa (where t is l or 2), -S(O)tRa (where t is l or 2) and -S(O)tN(Ra)2 (where t is l or 2) where each R21 is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), ?uoroalkyl, carbocyclyl (optionally substituted with halogen, y, methoxy, or tri?uoromethyl), carbocyclylalkyl (optionally substituted with halogen, y, methoxy, or tri?uoromethyl), aryl nally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), aralkyl (optionally tuted with halogen, hydroxy, methoxy, or tri?uoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), or arylalkyl (optionally substituted with halogen, y, metho xy, or tri?uoromethyl).
"Alkynyl" refers to a ht or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon triple bond, haVing from two to twelve carbon atoms. In n embodiments, an alkynyl comprises two to eight carbon atoms. In other embodiments, an alkynyl comprises two to six carbon atoms. In other ments, an alkynyl comprises two to four carbon atoms. The alkynyl is attached to the rest of the molecule by a single bond, for example, ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like. Unless stated otherwise speci?cally in the speci?cation, an alkynyl group is optionally substituted by one or more ofthe following tuents: halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, -ORa, - SRa, -OC(O)-Ra, -N(Ra)2, -C(O)Ra, -C(O)ORa, -C(O)N(Ra)2, -N(Ra)C(O)ORa, -OC(O)— N (Ra)2, -N(Ra)C(O)Ra, -N(Ra)S(O)tRa (where t is l or 2), ORa (where t is l or 2), -S(O)tRa (where t is l or 2) and -S(O)tN(Ra)2 (where t is l or 2) where each R21 is ndently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), ?uoroalkyl, carbocyclyl (optionally tuted with halogen, hydroxy, methoxy, or tri?uoromethyl), carbocyclylalkyl (optionally substituted with halogen, y, methoxy, or tri?uoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), heteroaryl nally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl).
"Alkylene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain g the rest of the molecule to a l group, consisting solely of carbon and hydrogen, containing no unsaturation and having ?om one to twelve carbon atoms, for example, methylene, ethylene, propylene, n-butylene, and the like. The alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the l group is through one carbon in the alkylene chain or through any two carbons within the chain. In n embodiments, an alkylene comprises one to eight carbon atoms (e.g., C1-C8 alkylene). In other embodiments, an alkylene comprises one to ?ve carbon atoms (e.g., C1-C5 alkylene). In other embodiments, an alkylene comprises one to four carbon atoms (e.g., C1-C4 ne). In other embodiments, an alkylene comprises one to three carbon atoms (e.g., C1-C3 alkylene). In other embodiments, an alkylene comprises one to two carbon atoms (e.g., C1-C2 alkylene). In other ments, an alkylene comprises one carbon atom (e.g., C1 alkylene). In other embodiments, an alkylene comprises ?ve to eight carbon atoms (e.g., C5-C8 alkylene). In other embodiments, an alkylene comprises two to ?ve carbon atoms (e. g., C2-C5 alkylene). In other embodiments, an alkylene comprises three to ?ve carbon atoms (e.g., C3-C5 alkylene). Unless stated otherwise speci?cally in the speci?cation, an alkylene chain is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, -ORa, - SRa, -OC(O)-Ra, 2, -C(O)Ra, -C(O)ORa, -C(O)N(Ra)2, -N(Ra)C(O)ORa, -OC(O)— N (Ra)2, -N(Ra)C(O)Ra, -N(Ra)S(O)tRa (where t is l or 2), -S(O)tORa (where t is l or 2), -S(O)tRa (where t is l or 2) and -S(O)tN(Ra)2 (where t is l or 2) where each R21 is independently hydrogen, alkyl (optionally tuted with n, hydroxy, methoxy, or tri?uoromethyl), ?uoroalkyl, yclyl (optionally substituted with halogen, hydroxy, y, or romethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), aryl nally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or romethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), heterocyclylalkyl (optionally substituted with n, hydroxy, methoxy, or tri?uoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl).
"Alkynylene" or "alkynylene chain" refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, ting solely of carbon and hydrogen, containing at least one carbon-carbon triple bond, and having ?om two to twelve carbon atoms. The alkynylene chain is attached to the rest ofthe molecule through a single bond and to the radical group through a single bond. In n embodiments, an lene comprises two to eight carbon atoms (e.g., C2-C8 alkynylene). In other embodiments, an lene comprises two to ?ve carbon atoms (e. g., C2-C5 alkynylene). In other embodiments, an alkynylene comprises two to four carbon atoms (e.g., C2-C4 alkynylene). In other embodiments, an alkynylene comprises two to three carbon atoms (e.g., C2-C3 alkynylene). In other embodiments, an alkynylene ses two carbon atom (e.g., C2 alkylene). In other embodiments, an alkynylene comprises ?ve to eight carbon atoms (e.g., C5-C8 alkynylene). In other embodiments, an alkynylene comprises three to ?ve carbon atoms (e.g., C3-C5 alkynylene). Unless stated otherwise speci?cally in the speci?cation, an alkynylene chain is optionally substituted by one or more ofthe following substituents: halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, -ORa, - SRa, -OC(O)-Ra, -N(Ra)2, -C(O)Ra, -C(O)ORa, -C(O)N(Ra)2, -N(Ra)C(O)ORa, -OC(O)— N (Ra)2, -N(Ra)C(O)Ra, -N(Ra)S(O)tRa (where t is l or 2), -S(O)tORa (where t is l or 2), -S(O)tRa (where t is l or 2) and -S(O)tN(Ra)2 (where t is l or 2) where each R21 is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), ?uoroalkyl, carbocyclyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), carbocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), aryl nally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), aralkyl (optionally tuted with halogen, hydroxy, methoxy, or tri?uoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or romethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, y, or tri?uoromethyl), heteroaryl (optionally substituted with halogen, y, methoxy, or tri?uoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl).
"Aryl" refers to a radical derived ?om an aromatic clic or yclic hydrocarbon ring system by removing a hydrogen atom ?om a ring carbon atom. The aromatic monocyclic or yclic arbon ring system contains only en and carbon ?om ?ve to eighteen carbon atoms, where at least one ofthe rings in the ring system is ?Jlly unsaturated, z'.e., it contains a , delocalized (4n+2) Tc—electron system in accordance with the Huckel theory. The ring system from which aryl groups are derived include, but are not d to, groups such as benzene, ?uorene, indane, indene, tetralin and naphthalene. Unless stated otherwise speci?cally in the speci?cation, the term "aryl" or the pre?x "ar-" (such as in "aralkyl") is meant to include aryl radicals optionally substituted by one or more substituents ndently selected from alkyl, l, alkynyl, halo, ?uoroalkyl, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted heteroarylalkyl, -Rb-ORa, (O)-Ra, -Rb-OC(O)-0Ra, -Rb-OC(O)-N(Ra)2, Ra) 2, -Rb-C(O)Ra, -Rb-C(O)0Ra, -Rb-C(O)N(Ra)2, -Rb-O-R°-C(O)N(Ra)2, -Rb-N(Ra)C(O)OR a, -Rb-N(Ra)C(O)Ra, -Rb-N(Ra)S(O)tRa (where t is 1 or 2), -Rb-S(O)tRa (where t is 1 or 2), -Rb-S(O)tORa (where t is l or 2) and -Rb-S(O)tN(Ra)2 (where t is l or 2), where each R81 is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, y, or tri?uoromethyl), ?uoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), cycloalkylalkyl (optionally substituted with halogen, y, methoxy, or tri?uoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), l (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or romethyl), heterocyclylalkyl (optionally substituted with n, hydroxy, methoxy, or tri?uoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or romethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), each Rb is independently a direct bond or a straight or branched alkylene or alkenylene chain, and Rc is a straight or branched alkylene or alkenylene chain, and where each of the above substituents is unsubstituted unless otherwise indicated.
"Aralkyl" refers to a radical of the formula -R°-aryl where R0 is an alkylene chain as de?ned above, for example, methylene, ethylene, and the like. The alkylene chain part of the aralkyl radical is optionally substituted as described above for an alkylene chain. The aryl part of the aralkyl radical is optionally substituted as bed above for an aryl group.
"Aralkenyl" refers to a radical of the formula —Rd-aryl where Rd is an alkenylene chain as de?ned above. The aryl part of the aralkenyl radical is optionally tuted as described above for an aryl group. The alkenylene chain part of the aralkenyl radical is optionally substituted as de?ned above for an alkenylene group.
"Aralkynyl" refers to a radical of the formula -Re-aryl, Where Re is an alkynylene chain as de?ned above. The aryl part of the aralkynyl radical is optionally substituted as described above for an aryl group. The alkynylene chain part of the aralkynyl radical is optionally substituted as de?ned above for an alkynylene chain. oxy" refers to a radical bonded through an oxygen atom ofthe a -O-R°—aryl where R0 is an alkylene chain as de?ned above, for example, methylene, ethylene, and the like. The alkylene chain part of the aralkyl radical is optionally substituted as described above for an alkylene chain. The aryl part of the aralkyl radical is optionally substituted as bed above for an aryl group.
"Carbocyclyl" refers to a stable non-aromatic monocyclic or clic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which includes ?lSGd or bridged ring systems, having from three to ?fteen carbon atoms. In certain embodiments, a carbocyclyl comprises three to ten carbon atoms. In other embodiments, a carbocyclyl comprises ?ve to seven carbon atoms. The carbocyclyl is attached to the rest of the molecule by a single bond. Carbocyclyl is saturated (z'.e., containing single C- C bonds only) or unsaturated (i.e., containing one or more double bonds or triple bonds).
A ?llly saturated carbocyclyl radical is also referred to as "cycloalkyl." Examples of clic cycloalkyls include, e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. An unsaturated carbocyclyl is also referred to as "cycloalkenyl." Examples of monocyclic cycloalkenyls include, e. g., entenyl, cyclohexenyl, eptenyl, and ctenyl. Polycyclic carbocyclyl radicals include, for example, adamantyl, norbomyl (i.e., bicyclo[2.2. l]heptanyl), norbomenyl, decalinyl, 7,7-dimethyl-bicyclo[2.2. l]heptanyl, and the like. Unless otherwise stated speci?cally in the speci?cation, the term "carbocyclyl" is meant to include carbocyclyl radicals that are optionally substituted by one or more substituents independently selected from alkyl, alkenyl, alkynyl, halo, ?uoroalkyl, oxo, thioxo, cyano, nitro, ally substituted aryl, optionally substituted aralkyl, optionally substituted nyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, ally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -Rb-ORa, (O)-Ra, (O)-0Ra, -Rb-OC(O)-N(Ra)2, Ra) 2, -Rb-C(O)Ra, -Rb-C(O)0Ra, -Rb-C(O)N(Ra)2, R°-C(O)N(Ra)2, -Rb-N(Ra)C(O)OR a, -Rb-N(Ra)C(O)Ra, -Rb-N(Ra)S(O)tRa (where t is 1 or 2), -Rb-S(O)tRa (where t is 1 or 2), -Rb-S(O)tORa (where t is l or 2) and -Rb-S(O)tN(Ra)2 (where t is l or 2), where each R81 is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), ?uoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), cycloalkylalkyl nally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), aryl (optionally substituted with n, hydroxy, methoxy, or tri?uoromethyl), aralkyl (optionally substituted with n, hydroxy, methoxy, or tri?uoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), heterocyclylalkyl (optionally tuted with halogen, hydroxy, methoxy, or tri?uoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), or arylalkyl (optionally substituted with halogen, hydroxy, methoxy, or romethyl), each Rb is independently a direct bond or a straight or branched alkylene or alkenylene chain, and Rc is a straight or branched alkylene or alkenylene chain, and where each of the above tuents is unsubstituted unless otherwise indicated.
"Carbocyclylalkyl" refers to a radical of the formula —R°-carbocyclyl where R0 is an alkylene chain as de?ned above. The alkylene chain and the carbocyclyl radical is optionally substituted as de?ned above.
"Carbocyclylalkynyl" refers to a radical of the formula —R°-carbocyclyl where R0 is an alkynylene chain as de?ned above. The alkynylene chain and the carbocyclyl radical is optionally substituted as de?ned above.
"Carbocyclylalkoxy" refers to a radical bonded h an oxygen atom of the formula —O-R°-carbocyclyl where R0 is an alkylene chain as de?ned above. The alkylene chain and the carbocyclyl l is optionally substituted as de?ned above.
As used herein, "carboxylic acid bioisostere" refers to a ?anctional group or moiety that exhibits similar physical, ical and/or chemical ties as a carboxylic acid moiety. Examples of carboxylic acid bioisosteres include, but are not limited to, O O N’N\ N’O A N’s ,OH A ,CN .m)\?wkmI ,N | >=o ‘N "EH «"LH "LEVER| i: E p8‘ 0 I N | \N lol ’ p ’ EQOH OH OH O and the like.
"Halo" or "halogen" refers to bromo, chloro, ?uoro or iodo substituents.
"Fluoroalkyl" refers to an alkyl radical, as de?ned above, that is substituted by one or more ?uoro ls, as de?ned above, for example, tri?uoromethyl, di?uoromethyl, ?uoromethyl, tri?uoroethyl, l-?uoromethyl?uoroethyl, and the like. In some embodiments, the alkyl part ofthe ?uoroalkyl radical is optionally substituted as de?ned above for an alkyl group.
"Heterocyclyl" refers to a stable 3- to 18-membered non-aromatic ring radical that comprises two to twelve carbon atoms and from one to six heteroatoms selected from nitrogen, oxygen and sul?lr. Unless stated otherwise speci?cally in the speci?cation, the heterocyclyl radical is a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which ally includes ?ased or bridged ring systems. The heteroatoms in the heterocyclyl radical are optionally oxidized. One or more nitrogen atoms, if present, are ally quatemized. The heterocyclyl radical is partially or ?llly saturated. The heterocyclyl is attached to the rest of the molecule through any atom of the ring(s). Examples of such heterocyclyl radicals include, but are not limited to, dioxolanyl, thienyl[l,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, azolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, lidinyl, thiazolidinyl, tetrahydro?lryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, l-oxo-thiomorpholinyl, and 1,l-dioxo-thiomorpholinyl. Unless stated otherwise speci?cally in the speci?cation, the term "heterocyclyl" is meant to include heterocyclyl radicals as de?ned above that are optionally substituted by one or more substituents selected from alkyl, alkenyl, alkynyl, halo, lkyl, oxo, thioxo, cyano, nitro, optionally tuted aryl, optionally substituted aralkyl, ally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -Rb-ORa, -Rb-OC(O)-Ra, -Rb-OC(O)-0Ra, -Rb-OC(O)-N(Ra)2, -Rb-N(Ra) 2, -Rb-C(O)Ra, -Rb-C(O)0Ra, -Rb-C(O)N(Ra)2, -Rb-O-R°-C(O)N(Ra)2, -Rb-N(Ra)C(O)OR a, -Rb-N(Ra)C(O)Ra, -Rb-N(Ra)S(O)tRa (where t is 1 or 2), -Rb-S(O)tRa (where t is 1 or 2), -Rb-S(O)tORa (where t is l or 2) and -Rb-S(O)tN(Ra)2 (where t is l or 2), where each R81 is independently en, alkyl (optionally tuted with halogen, hydroxy, methoxy, or tri?uoromethyl), ?uoroalkyl, cycloalkyl nally tuted with halogen, hydroxy, methoxy, or tri?uoromethyl), cycloalkylalkyl (optionally substituted with n, hydroxy, y, or tri?uoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or romethyl), l (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), heterocyclylalkyl (optionally substituted with halogen, y, methoxy, or tri?uoromethyl), heteroaryl nally tuted with halogen, hydroxy, methoxy, or tri?uoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), each Rb is independently a direct bond or a straight or branched alkylene or alkenylene chain, and Rc is a straight or branched alkylene or alkenylene chain, and Where each of the above tuents is unsubstituted unless otherwise indicated.
"N—heterocyclyl" or "N-attached heterocyclyl" refers to a heterocyclyl l as de?ned above containing at least one nitrogen and Where the point of attachment of the heterocyclyl radical to the rest of the molecule is through a nitrogen atom in the heterocyclyl radical. An N-heterocyclyl radical is optionally substituted as described above for heterocyclyl radicals. Examples of such N—heterocyclyl radicals include, but are not limited to, l-morpholinyl, l-piperidinyl, l-piperazinyl, l-pyrrolidinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl.
"C—heterocyclyl" or "C-attached heterocyclyl" refers to a heterocyclyl radical as de?ned above ning at least one heteroatom and Where the point of attachment of the heterocyclyl radical to the rest of the molecule is through a carbon atom in the heterocyclyl radical. A C—heterocyclyl radical is optionally substituted as described above for heterocyclyl radicals. Examples of such C—heterocyclyl radicals e, but are not limited to, 2-morpholinyl, 2- or 3- or 4-piperidinyl, 2-piperazinyl, 2- or 3- pyrrolidinyl, and the like.
"Heterocyclylalkyl" refers to a radical of the formula —R°-heterocyclyl where R0 is an ne chain as de?ned above. If the heterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl is ally attached to the alkyl radical at the nitrogen atom. The alkylene chain ofthe heterocyclylalkyl radical is optionally substituted as de?ned above for an alkylene chain. The heterocyclyl part ofthe heterocyclylalkyl radical is optionally substituted as de?ned above for a cyclyl group.
"Heterocyclylalkoxy" refers to a radical bonded through an oxygen atom of the formula —O-R°—heterocyclyl where R0 is an alkylene chain as de?ned above. If the heterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl is optionally attached to the alkyl radical at the nitrogen atom. The alkylene chain ofthe heterocyclylalkoxy radical is optionally substituted as de?ned above for an alkylene chain. The cyclyl part of the heterocyclylalkoxy radical is ally substituted as de?ned above for a heterocyclyl group.
"Heteroaryl" refers to a radical derived from a 3- to 18-membered aromatic ring radical that comprises two to seventeen carbon atoms and from one to six heteroatoms selected from nitrogen, oxygen and sulfur. As used , the heteroaryl l is a clic, bicyclic, tricyclic or tetracyclic ring system, wherein at least one ofthe rings in the ring system is ?llly unsaturated, z'.e., it contains a cyclic, delocalized (4n+2) Tc—electron system in accordance with the Huckel . Heteroaryl includes fused or bridged ring systems. The heteroatom(s) in the aryl radical is optionally oxidized. One or more nitrogen atoms, if present, are optionally quatemized. The heteroaryl is attached to the rest of the molecule through any atom of the ring(s).
Examples of heteroaryls include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3 dioxolyl, benzo?lranyl, benzooxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][l,4]dioxepinyl, benzo[b][l,4]oxazinyl, l,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzo?lranyl, benzo?lranonyl, benzothienyl (benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[l,2-a]pyridinyl, olyl, cinnolinyl, cyclopenta[d]pyrimidinyl, 6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl, ,6-dihydrobenzo [h] quinazolinyl, 5,6-dihydrobenzo [h] cinno linyl, hydro-5H- benzo[6,7]cyclohepta[l,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, ?aranyl, ?lranonyl, ?Jro[3,2-c]pyridinyl, 5,6,7,8,9,l0-hexahydrocycloocta[d]pyrimidinyl, ,6,7, 8,9, l 0-hexahydrocycloocta[d]pyridazinyl, ,6,7,8,9,l0-hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, nyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, 8-methano-5 ,6,7, 8-tetrahydroquinazo linyl, naphthyridinyl, l ,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, yl, ,7,8,9,lO,lOa-octahydrobenzo[h]quinazolinyl, l-phenyl-lH—pyrrolyl, inyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazo lo [3 ,4-d]pyrimidinyl, nyl, pyrido [3 ,2 -d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, ,6,7, 8-tetrahydroquinazo linyl, 5 ,6,7, 8 -tetrahydrobenzo [4,5 ]thieno [2,3 -d]pyrimidinyl, 6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl, ,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, thieno [2,3-d]pyrimidinyl, thieno [3 yrimidinyl, thieno [2,3 -c]pridinyl, and thiophenyl (z'.e. thienyl). Unless stated otherwise specifically in the specification, the term "heteroaryl" is meant to include heteroaryl radicals as de?ned above which are optionally substituted by one or more tuents selected from alkyl, alkenyl, alkynyl, halo, ?uoroalkyl, haloalkenyl, haloalkynyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, ally tuted aralkyl, ally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted yclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally tuted heteroaryl, optionally substituted heteroarylalkyl, -Rb-ORa, -Rb-OC(O)-Ra, -Rb-OC(O)-0Ra, -Rb-OC(O)-N(Ra)2, -Rb-N(Ra) 2, -Rb-C(O)Ra, -Rb-C(O)0Ra, -Rb-C(O)N(Ra)2, R°-C(O)N(Ra)2, -Rb-N(Ra)C(O)OR a, -Rb-N(Ra)C(O)Ra, -Rb-N(Ra)S(O)tRa (where t is 1 or 2), -Rb-S(O)tRa (where t is 1 or 2), -Rb-S(O)tORa (where t is l or 2) and -Rb-S(O)tN(Ra)2 (where t is l or 2), where each R81 is independently hydrogen, alkyl (optionally substituted with halogen, y, methoxy, or tri?uoromethyl), ?uoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or romethyl), l (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or tri?uoromethyl), each Rb is independently a direct bond or a straight or branched alkylene or lene chain, and Rc is a ht or branched ne or alkenylene chain, and where each of the above substituents is unsubstituted unless otherwise indicated.
"N—heteroaryl" refers to a heteroaryl radical as de?ned above containing at least one nitrogen and where the point of attachment of the heteroaryl radical to the rest ofthe molecule is through a en atom in the heteroaryl radical. An N—heteroaryl radical is optionally substituted as described above for heteroaryl radicals.
"C—heteroaryl" refers to a heteroaryl radical as de?ned above and where the point of attachment of the heteroaryl radical to the rest of the molecule is through a carbon atom in the heteroaryl radical. A C-heteroaryl radical is optionally substituted as described above for heteroaryl radicals.
"Heteroarylalkyl" refers to a l of the formula —R°-heteroaryl, where R0 is an alkylene chain as de?ned above. If the heteroaryl is a nitrogen-containing heteroaryl, the heteroaryl is optionally attached to the alkyl radical at the nitrogen atom.
The alkylene chain ofthe heteroarylalkyl radical is optionally substituted as de?ned above for an alkylene chain. The heteroaryl part of the heteroarylalkyl radical is ally substituted as de?ned above for a heteroaryl group.
"Heteroarylalkoxy" refers to a l bonded through an oxygen atom of the formula —O-R°—heteroaryl, where R0 is an ne chain as de?ned above. If the heteroaryl is a nitrogen-containing heteroaryl, the heteroaryl is optionally attached to the alkyl radical at the nitrogen atom. The alkylene chain ofthe heteroarylalkoxy radical is optionally substituted as de?ned above for an alkylene chain. The heteroaryl part of the heteroarylalkoxy radical is optionally tuted as de?ned above for a heteroaryl group.
The compounds disclosed herein, in some embodiments, contain one or more asymmetric s and thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that are de?ned, in terms of absolute stereochemistry, as (R)- or (S)-. Unless stated otherwise, it is intended that all stereoisomeric forms ofthe compounds disclosed herein are contemplated by this disclosure. When the compounds described herein contain alkene double bonds, and unless speci?ed otherwise, it is intended that this disclosure includes both E and Z geometric isomers (e.g., cis or trans.) Likewise, all possible isomers, as well as their racemic and optically pure forms, and all tautomeric forms are also intended to be included. The term tric isomer" refers to E or Z geometric s (e.g., cis or trans) of an alkene double bond. The term "positional isomer" refers to structural isomers around a central ring, such as 0rth0-, meta-, and para- isomers around a benzene ring.
A "tautomer" refers to a molecule wherein a proton shift from one atom of a molecule to another atom ofthe same molecule is possible. The compounds presented , in certain embodiments, exist as tautomers. In stances where tautomerization is le, a chemical equilibrium of the tautomers will exist. The exact ratio of the tautomers depends on several factors, including physical state, ature, solvent, and pH. Some examples oftautomeric equilibrium include: XANH2=K NH \kN/‘X?x ")1 N‘m’ NN’N HN\,\]' NrN'NH AIKEN ‘11: H | \>—E __ \[N A A lN/>—§ E/ N / NH NH VOHI _ §_MO "Pharmaceutically able salt" includes both acid and base addition salts. A ceutically acceptable salt ofany one ofthe substituted cyclic derivative compounds described herein is intended to ass any and all pharmaceutically suitable salt forms. Preferred pharmaceutically acceptable salts of the compounds described herein are pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. aceutically acceptable acid addition salt" refers to those salts which retain the biological effectiveness and properties ofthe free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sul?lric acid, nitric acid, phosphoric acid, hydroiodic acid, hydro?uoric acid, phosphorous acid, and the like. Also included are salts that are formed with organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and. aromatic ic acids, etc. and include, for example, acetic acid, tri?uoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, c acid, tartaric acid, citric acid, benzoic acid, ic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Exemplary salts thus include sulfates, pyrosulfates, bisulfates, sul?tes, bisul?tes, nitrates, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, s, acetates, tri?uoroacetates, propionates, caprylates, isobutyrates, oxalates, malonates, succinate suberates, sebacates, ?Jmarates, maleates, mandelates, benzoates, benzoates, methylbenzoates, dinitrobenzoates, phthalates, benzenesulfonates, esulfonates, phenylacetates, citrates, lactates, malates, tartrates, methanesulfonates, and the like. Also contemplated are salts ofamino acids, such as arginates, gluconates, and galacturonates (see, for example, Berge S.M. et al., "Pharmaceutical Salts," Journal ofPharmaceutical Science, 66: 1-19 (1997)). Acid addition salts ofbasic compounds are, in some embodiments, prepared by contacting the free base forms with a ent amount ofthe desired acid to produce the salt according to methods and techniques with which a skilled artisan is familiar.
"Pharmaceutically acceptable base addition salt" refers to those salts that retain the biological effectiveness and properties ofthe free acids, which are not biologically or otherwise rable. These salts are prepared from addition of an inorganic base or an organic base to the free acid. Pharmaceutically acceptable base addition salts are, in some embodiments, formed with metals or amines, such as alkali and ne earth metals or organic amines. Salts derived from inorganic bases include, but are not d to, sodium, potassium, lithium, ammonium, m, ium, iron, zinc, copper, manganese, aluminum salts and the like. Salts derived from organic bases include, but are not d to, salts ofprimary, secondary, and tertiary amines, substituted amines including lly occurring substituted amines, cyclic amines and basic ion exchange resins, for example, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, nolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, ine, caffeine, procaine, N,N— dibenzylethylenediamine, chloroprocaine, hydrabamine, choline, betaine, ethylenediamine, ethylenedianiline, N—methylglucamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N—ethylpiperidine, polyamine resins and the like. See Berge et al., supra.
As used herein, "treatment" or "treating," or "palliating" or "ameliorating" are used interchangeably. These terms refer to an approach for obtaining bene?cial or desired results including but not limited to therapeutic bene?t and/or a prophylactic bene?t. By "therapeutic bene?t" is meant eradication or amelioration of the ying disorder being treated. Also, a therapeutic bene?t is achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the patient, notwithstanding that the patient is still ed with the ying er. For lactic , the compositions are, in some embodiments, administered to a patient at risk of developing a particular disease, or to a patient reporting one or more of the physiological symptoms of a disease, even though a diagnosis of this disease has not been made. ug" is meant to indicate a compound that is, in some embodiments, converted under physiological conditions or by solvolysis to a biologically active compound described herein. Thus, the term "prodrug" refers to a precursor of a biologically active compound that is pharmaceutically acceptable. A prodrug is typically inactive when administered to a subject, but is converted in vivo to an active compound, for example, by hydrolysis. The prodrug compound often offers advantages of solubility, tissue compatibility or delayed e in a mammalian organism (see, e.g., Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam).
A discussion ofprodrugs is provided in Higuchi, T., et al., "Pro-drugs as Novel Delivery s," A.C.S. Symposium Series, Vol. 14, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987.
The term "prodrug" is also meant to include any ntly bonded carriers, which e the active compound in vivo when such prodrug is administered to a mammalian subject. Prodrugs of an active compound, as bed herein, are prepared by modifying ?anctional groups present in the active compound in such a way that the ations are cleaved, either in routine manipulation or in vivo, to the parent active compound. Prodrugs include compounds wherein a hydroxy, amino or mercapto group is bonded to any group that, when the prodrug of the active compound is administered to a mammalian subject, s to form a free hydroxy, free amino or free mercapto group, respectively. Examples rugs e, but are not limited to, acetate, formate and benzoate tives of alcohol or amine ?anctional groups in the active compounds and the like.
Substituted Heterocyclic Derivative Compounds Substituted cyclic derivative compounds are described herein that are lysine speci?c demethylase-l inhibitors. These compounds, and compositions comprising these compounds, are useful for the treatment of cancer and neoplastic disease.
One embodiment provides a compound having the structure of Formula (I), or a pharmaceutically acceptable salt thereof, wherein, W is N, C-H, or C-F; X is hydrogen, halogen, -CN, optionally substituted alkyl, optionally substituted alkynyl, optionally substituted carbocyclylalkynyl, ally substituted aryl, or optionally substituted heteroaryl; Y is hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, or optionally substituted cycloalkylalkyl; Z is an ally substituted group chosen from alkyl, carbocyclyl, C-attached heterocyclyl, N-attached cyclyl, heterocyclylalkyl, heterocyclylalkenyl, -O- heterocyclyl, -N(R)-heterocyclyl, -O-heterocyclylalkyl, -N(R)-heterocyclylalkyl, - N(R)(C1-C4alkylene)-NR2, C4alkylene)-NR2, and R is en or C1-C4alkyl.
One embodiment provides a nd ofFormula (I) having the structure of a (Ia), or a pharmaceutically acceptable salt thereof, W is N, C-H, or C-F; X is hydrogen, halogen, -CN, optionally substituted alkynyl, optionally substituted carbocyclylalkynyl, optionally substituted aryl, or optionally substituted heteroaryl; Y is hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, or optionally tuted cycloalkylalkyl; and Z is an optionally substituted group chosen from N—attached heterocyclyl, -O- heterocyclylalkyl, -N(H)-heterocyclyl, -N(Me)-heterocyclyl, -N(H)-heterocyclylalkyl, or -N(Me)-heterocyclylalkyl.
One embodiment provides a compound ofFormula (I) or (Ia) having the structure of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein, W is N, C-H, or C-F; X is hydrogen, halogen, optionally substituted alkynyl, optionally tuted carbocyclylalkynyl, optionally substituted aryl, or optionally substituted heteroaryl; Y is hydrogen, optionally substituted alkyl, or optionally substituted cycloalkyl; Z is an optionally substituted group chosen from N—heterocyclyl, -O- heterocyclylalkyl, -N(H)-heterocyclylalkyl, or -N(Me)-heterocyclylalkyl.
Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, n W is C-H. Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein W is C-F. Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein W is N. r embodiment provides the compound of Formula (lb), or a ceutically acceptable salt thereof, wherein X is hydrogen. Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein X is halogen. r embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein X is optionally substituted alkynyl.
Another embodiment provides the compound of a (lb), or a ceutically acceptable salt thereof, wherein X is optionally substituted carbocyclylalkynyl.
Another embodiment provides the compound of Formula (lb), or a ceutically acceptable salt thereof, wherein X is optionally substituted aryl, or optionally substituted heteroaryl. Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein X is optionally substituted aryl. Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein X is an optionally substituted phenyl.
Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein X is ally substituted aryl. Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, n X is chosen from an ally tuted pyridinyl, optionally substituted pyrazolyl, or optionally substituted indazolyl.
Another embodiment provides the compound of Formula (lb), or a pharmaceutically able salt f, wherein Y is hydrogen. r embodiment provides the compound of Formula (lb), or a ceutically acceptable salt thereof, wherein Y is optionally substituted cycloalkyl. Another embodiment provides the compound ofFormula (lb), or a pharmaceutically acceptable salt f, wherein Y is optionally substituted alkyl. Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein Y is an optionally substituted C1-C3 alkyl. Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, n Y is an optionally substituted C1 alkyl. r embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein Y is a methyl group.
Another ment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein Z is an optionally substituted heterocyclylalkyl. Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein Z is an optionally substituted -N(H)- heterocyclylalkyl. Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein Z is an optionally substituted -N(Me)- heterocyclylalkyl.
Another embodiment provides the compound of a (lb), or a pharmaceutically acceptable salt thereof, n Z is an optionally substituted heterocyclylalkyl and the heterocyclylalkyl group has the formula —R°-heterocyclyl and the Rc is an optionally substituted C1-C3 alkylene chain. Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, n Z is an optionally substituted -O-heterocyclylalkyl and the heterocyclylalkyl group has the a —R°-heterocyclyl and the Rc is an optionally substituted C1 alkylene chain.
Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein Z is an optionally substituted cyclylalkyl and the heterocyclylalkyl group has the formula —R°-heterocyclyl and the heterocyclyl is an optionally substituted nitrogen-containing 4-, 5-, 6-, or 7- membered cyclyl.
Another embodiment es the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein Z is an optionally substituted -N(H)- heterocyclylalkyl and the heterocyclylalkyl group has the formula —R°-heterocyclyl and the Rc is an optionally substituted C1-C3 alkylene chain. Another embodiment es the nd of a (lb), or a pharmaceutically acceptable salt f, wherein Z is an optionally substituted -N(H)-heterocyclylalkyl and the heterocyclylalkyl group has the formula —R°-heterocyclyl and the Rc is an ally substituted C1 alkylene chain.
Another embodiment provides the compound of Formula (lb), or a ceutically acceptable salt thereof, wherein Z is an optionally substituted -N(H)- heterocyclylalkyl and the heterocyclylalkyl group has the formula —R°-heterocyclyl and the heterocyclyl is an optionally substituted nitrogen-containing 4-, 5-, 6-, or 7- membered heterocyclyl.
Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein Z is an optionally substituted -N(Me)- heterocyclylalkyl and the heterocyclylalkyl group has the formula —R°-heterocyclyl and the Rc is an ally substituted C1-C3 alkylene chain. Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein Z is an optionally tuted -N(Me)-heterocyclylalkyl and the heterocyclylalkyl group has the formula —R°-heterocyclyl and the Rc is an optionally substituted C1 alkylene chain.
Another embodiment provides the compound of a (lb), or a pharmaceutically acceptable salt f, wherein Z is an optionally substituted -N(Me)- heterocyclylalkyl and the heterocyclylalkyl group has the a —R°-heterocyclyl and the heterocyclyl is an optionally substituted nitrogen-containing 4-, 5-, 6-, or 7- membered heterocyclyl.
Another embodiment provides the compound of Formula (lb), or a ceutically acceptable salt thereof, wherein Z is an optionally substituted N- heterocyclyl. Another embodiment provides the compound of Formula (lb), or a ceutically acceptable salt thereof, wherein Z is a 4-, 5-, 6-, or 7-membered N- heterocyclyl. Another embodiment provides the compound of Formula (lb), or a pharmaceutically acceptable salt thereof, wherein Z is a 6-membered N—heterocyclyl.
Another embodiment provides the compound of Formula (lb), or a ceutically acceptable salt thereof, wherein Z is an optionally substituted piperidine. r ment provides the compound of Formula (lb), or a pharmaceutically able salt thereof, wherein Z is an optionally substituted 4-aminopiperidine.
In some embodiments, the substituted heterocyclic derivative compound described in a (I), (la), or (Ib) has a structure provided in Table l.
TABLE 1 4-(2-(4-amin0piperidiny1) methyl0x0p-t01y1-1,6- dihydropyrimidiny1)benz0nitrile 4-[2-(4-amin0-piperidiny1)-5 -(4- methoxy—pheny1)methy10x0-1,6- dihydro—pyrimidiny1]-benzonitrile 4-[2-(4-amin0-piperidiny1)(6- methoxy—pyridin-3 -methy1 OX0-1,6-dihyd1‘O-py1‘imidiny1]- benzonitrile 4-[2-(4-amin0-piperidiny1) methyl(6-methy1-pyridin-3 -y1) OX0-1,6-dihyd1‘O-py1‘imidiny1]- benzonitrile 4-[2-(4-amin0-piperidiny1)-5 -(4- methoxy—pheny1)methy10x0-1,6- dihydro—pyrimidiny1]-benzonitrile 4-[2-(4-amin0-piperidiny1)-5 -(4- methoxy—pheny1)methy10x0-1,6- dihydro—pyrimidiny1]?u0r0- benzonitrile 4-[2-(4-amin0-piperidiny1)(3 - ?u0r0meth0xy-phenyl)methy1 OXO-l,6-dihyd1‘O-py1‘imidiny1] ?uorO-benzonitrile 4-[2-(4-amin0-piperidiny1)(6- methoxy—pyridin-3 -y1)methy1 OXO-l,6-dihyd1‘O-py1‘imidiny1] ?uorO-benzonitrile 4-[2-(4-amin0-piperidiny1)(6- methoxy—pyridin-3 -y1)methy1 OXO-l,6-dihyd1‘O-py1‘imidiny1] ?uorO-benzonitrile 4-[2-(4-amin0-piperidiny1)(6- ethyl-pyridin-3 -y1)methy10X0- 1,6-dihydr0-pyrimidiny1] - benzonitrile 2-?u0r0 [5-(4-meth0xy—pheny1) 11 methyl(4-methy1amin0-piperidin y1)0x0-1,6-dihydr0-pyrimidin y1]-benzonitrile 2-?u0r0 [5 -(3 -?u0r0meth0xy— 2 )methy1(4-methy1amin0- piperidin-l-y1)0x0-1,6-dihydr0- pyrimidiny1] -benzonitrile 4-amin0-piperidiny1)ethy1- 3 6-0X0- 1 ,6-dihydr0-pyrimidiny1] ?uorO-benzonitrile 4-[2-(4-amin0-piperidiny1)-5 - 4 cyclopentylethynyl- 1 10x0- 1,6-dihydr0-pyrimidiny1]?u0r0- benzonitrile [2-(4-amin0-piperidiny1)(4- cyan0-3 -?u0r0-pheny1)(4-meth0xy— phenyl)0x0-6H-pyrimidiny1]— acetic acid 2-[2-(4-amin0-piperidiny1)(4- 6 cyan0-3 -?u0r0-pheny1)(4-meth0xy— phenyl)0x0-6H-pyrimidiny1]— acetamide 4-amin0-piperidiny1)(3 - 7 hydroxy—propy1)0x0-1,6-dihydr0- pyrimidiny1]?u0r0-benzonitrile 4-[2-(4-amin0-piperidiny1)-5 - 8 benzo?lran-S-yl-l-methy10X0-1,6- dihydro—pyrimidiny1]?u0r0- benzonitrile 2-(4-amin0-piperidiny1)(4- 9 cyan0-3 -?u0r0-pheny1)methy1 OXO-l,6-dihyd1‘O-py1‘imidine-5 - carbonitrile 4-[2-(4-amin0piperidiny1)-5 -ch10r0- 1-methy10X0pyrimidiny1] ?uorobenzonitrile 2-?u0r0[1-methy1(4- 2 methylamino—piperidiny1)-5 -(6- methyl-pyridiny1)0x0-1,6- dihydro—pyrimidiny1]-benzonitrile 4-[2-(2,8-diaza-spir0[4.5]dec-8—y1) 22 (3 -?u0r0meth0xy-pheny1) methyl0X0-1,6-dihydr0-pyrimidin- 4-y1] ?u0r0-benzonitrile 4- {2-(4-amin0piperidyl)methy1 23 0X0-5 - [6-(tri?u0r0methy1) (3 y1)] hydropyrimidiny1} enzenecarbonitrile 4-[2-(4-amin0piperidyl)methy1 24 (2-methy1(2H-indazol-5 -y1)) 0x0hydropyrimidin yl]benzenecarb0nitrile 4-[2-((3R)-3 -amin0piperidyl)(3 - ?u0r0meth0xypheny1)- 1 1 0X0hydropyrimidiny1]—2- ?uorobenzenecarbonitrile 4- [2-(4-amin0piperidyl) -5 -(5 -?u0r0 26 methoxy(3 -5,6-dihydr0pyridy1)) methyl0x0hydr0pyrimidiny1] enzenecarbonitrile 4-[2-((3R)-3 -amin0pyrrolidiny1)(3 - 27 ?u0r0meth0xypheny1)methy1 0X0hydropyrimidiny1]—2- ?uorobenzenecarbonitrile 4-[2-((3 S)amin0-piperidiny1) 28 (3 -?u0r0meth0xy-pheny1) methyl0X0-1,6-dihydr0-pyrimidin- 4-y1] r0-benzonitrile 4-[2-((3 min0-pyrrolidiny1) 29 (3 -?u0r0meth0xy-pheny1) methyl0X0-1,6-dihydr0-pyrimidin- 4-y1] ?u0r0-benzonitrile 4-[2-((3R)-3 -amin0piperidyl)(4- methoxyphenyl) methy10X0hydr0 pyrimidiny1] ?uorobenzenecarbonitrile 4-[2-((3 S)amin0-piperidiny1) 3 (4-meth0xy—phenyl)methy10X0- 1,6-dihydr0-pyrimidiny1]?u0r0- benzonitrile 4- [2-(4-amino—4-methy1-piperidin 32 y1)-5 -(3 meth0xy—pheny1) methyl0X0-1,6-dihydr0-pyrimidin- 4-y1] ?u0r0-benzonitrile 4-[2-(4-amin0piperidy1)methy1-5 - 33 (1 -methy1(1H-indazol-5 -y1)) ropyrimidin yl]benzenecarb0nitrile 4- {2-(4-amin0-piperidiny1) 34 methyl0x0[1-(2,2,2-tri?u0r0- ethy1)- az01y1]— 1 ,6-dihydr0- pyrimidiny1} ?u0r0-benzonitrile 4-[2-(4-amin0-piperidiny1) methyl(1-methy1-1H-indazol-5 -y1)- 6-0X0- 1 ,6-dihydr0-pyrimidiny1] ?uorO-benzonitrile 4- {2-(4-amin0-piperidiny1) 36 methyl0x0[1-(2,2,2-tri?u0r0- ethy1)- 1H-pyraz01y1]— 1 ,6-dihydr0- pyrimidiny1} -benzonitrile 4-[2-(4-amin0piperidyl)methy1 37 (2-methy1(2H-indazol-5 -y1)) 0X0hydropyrimidiny1]—2- ?uorobenzenecarbonitrile 4-[2-(4-amin0piperidy1)(3,5- 38 di?u0r0meth0xypheny1)methyl- ydr0pyrimidin zenecarb0nitrile 4- [2-(4-amin0piperidy1)(4-cyan0-3 - 39 ?uoropheny1)-3 -methy10x0-3 - hydropyrimidin-S-y1]benzoic acid {4- [2-(4-amin0piperidy1)(4- 40 cyanopheny1)-3 -methy10x0(3 -hydr0 pyrimidin-S-y1)]?u0r0pheny1}-N— methylcarboxamide 4- [2-(4-amin0piperidy1)(4- 4 cyanopheny1)-3 -methy10x0(3 -hydr0 pyrimidin-S -y1)]?u0r0benzamide 4-[2-(4-amin0-piperidiny1) 42 methyl0x0(1-0x0-2,3 -dihydr0- 1H-isoind01y1)-1,6-dihydr0- pyrimidiny1]?u0r0-benzonitrile 3 -[2-(4-amin0-piperidiny1)(4- 43 cyan0-3 -?u0r0-pheny1)methy1 OXO-1,6-dihyd1‘0-py1‘imidi1’l-5 -y1] - benzoic acid 4- {5 -(3 -?u0r0meth0xy-pheny1) 44 methyl0x0[(3 S)-(pyrrolidin-3 - ylmethy1)-amin0]- 1 ,6-dihydr0- pyrimidiny1} -benzonitrile 4- {5 -(3 meth0xy-pheny1) 45 methyl0x0[(3R)-(pyrrolidin-3 - ylmethy1)-amin0]- 1 ,6-dihydr0- diny1} -benzonitrile 4-[2-[1,4]diazepany1(3 46 methoxy—pheny1)methy10x0-1,6- dihydro—pyrimidiny1]?u0r0- benzonitrile 2-?u0r0 [5 -(3 -?u0r0meth0xy— 47 pheny1)methy10x0piperazin- 1-y1-1,6-dihydr0-pyrimidinyl]- benzonitrile 4- [5 -(3 -?u0r0meth0xy-pheny1) 48 methyl0x0(piperidiny1amin0)- 1,6-dihydr0-pyrimidiny1] - benzonitrile 4-[2-(4-amin0-piperidiny1)-2'— 49 dimethylamino—1-methy10x0-1,6- dihydro— [5 ,5 ']bipyrimidiny1y1] ?uorO-benzonitrile -[2-(4-amin0-piperidiny1)(4- cyan0-3 -pheny1)methy1 50 OXO-1,6-dihyd1‘0-py1‘imidin-5 -y1] - pyridine-Z-carboxylic acid methylamide 0 meth0xy—pheny1) methyl0x0[(3 S)-(pyrrolidin-3 - ylmethy1)-amin0] - 1 ,6-dihydr0- pyrimidiny1} -benzonitrile 2-?u0r0 {5-(4-meth0xy—pheny1) 52 methyl0x0[(3R)-(pyrrolidin-3 - ylmethy1)-amin0]- 1 ,6-dihydr0- pyrimidiny1} -benzonitrile 2-?u0r0 [5-(4-meth0xy—pheny1) 53 methyl0x0(piperidiny1amin0)- 1,6-dihydr0-pyrimidiny1] - benzonitrile 2-?u0r0 [5-(4-meth0xy—pheny1) 54 methyl(methy1-(3 S)-pyrrolidin-3 - ylmethyl-amin0)0X0- 1 ,6-dihydr0- pyrimidiny1] -benzonitrile 2-?u0r0 [5-(4-meth0xy—pheny1) 55 methyl(methy1-piperidiny1- 0X0- 1 ,6-dihydr0-pyrimidin- 4-y1] -benzonitrile 2-?u0r0 [5-(4-meth0xy—pheny1) 56 methyl(methy1-pyrrolidin-3 - ylmethyl-amin0)0X0- 1 ,6-dihydr0- pyrimidiny1] -benzonitrile 4-[2-(4-amin0-piperidiny1)(6- 57 dimethylamino—pyridin-3 -y1) methyl0X0-1,6-dihydr0-pyrimidin- 4-y1] ?u0r0-benzonitrile 2-?u0r0 [5-(6-meth0xy—pyridin-3 - 58 y1)methy1(4-methy1amin0- piperidin-l-y1)0x0-1,6-dihydr0- diny1] nitrile 4-[2-(4-amin0-piperidiny1)-5 -(4- 59 dimethylamino—phenyl)methy1 OXO-l,6-dihyd1‘O-py1‘imidiny1] ?uorO-benzonitrile 4-amin0-piperidiny1) methyl0x0(6-pyrrolidiny1- pyridin-3 -y1)-1,6-dihydr0-pyrimidin y1] ?uoro—benzonitrile 4-[2-[1,4]diazepany1(6-meth0xy— 61 pyridin-3 -y1)methy10x0-1,6- dihydro—pyrimidiny1]?u0r0- benzonitrile 4-[2-[1,4]diazepany1(6-meth0xy— 62 pyridin-3 -y1)methy10x0-1,6- dihydro—pyrimidiny1]?u0r0- benzonitrile 4-[2-[1,4]diazepany1(6- 63 dimethylamino—pyridin-3 -y1) methyl0X0-1,6-dihydr0-pyrimidin- 4-y1] ?u0r0-benzonitrile 4-[2-(3 -amin0-azetidiny1)(4- 64 methoxy—pheny1)methy10x0-1,6- dihydro—pyrimidiny1]?u0r0- benzonitrile 2-?u0r0[1-methy1(4- 65 methylamino—piperidiny1)-5 -(2- methyl-ZH-indazol-S-y1)0X0-1,6- o—pyrimidiny1]-benzonitrile 4-[2-[1,4]diazepany1methy1 (2-methy1-2H-indazoly1)0X0-1,6- dihydro—pyrimidiny1]?u0r0- benzonitrile 4-[2-[1,4]diazepany1(6- 67 dimethylamino—pyridin-3 -y1) 0X0-1,6-dihydr0-pyrimidin- 4-y1] -benzonitrile 4-[2-(4-amin0-piperidiny1) methyl(6-m0rpholiny1-pyridin-3 - y1)0x0-1,6-dihydr0-pyrimidin y1] ?uoro—benzonitrile 4- [2-(3 -amin0methy1-azetidiny1)-5 - (4-meth0xy—phenyl)methy10X0- 1,6-dihydr0-pyrimidiny1]?u0r0- benzonitrile 2-?u0r0 [5-(4-meth0xy—pheny1) 70 methyl(3 -methy1amin0methy1- azetidin-l-y1)0x0-1,6-dihydr0- pyrimidiny1] -benzonitrile 4- [2-(4-dimethylamino—piperidin 7 y1)methy1-5 -(2-methy1-2H-indazoly1)0X0-1,6-dihydr0-pyrimidin y1] ?uoro—benzonitrile 4- dimethylamino—piperidin 72 y1)methy1(1-methy1-1H-indazoly1)0X0-1,6-dihydr0-pyrimidin y1] ?uoro—benzonitrile 4-amin0-piperidiny1)(1H- 73 ind01y1)methy10x0-1,6- dihydro—pyrimidiny1]?u0r0- benzonitrile 4-amin0-piperidiny1) 74 methyl-S-(l-methy1-1H-ind01y1) OXO-l,6-dihyd1‘O-py1‘imidiny1] ?uorO-benzonitrile 4-[2-(4-amin0-piperidiny1)(1H- 75 ind01y1)methy10x0-1,6- dihydro—pyrimidiny1]?u0r0- benzonitrile 4-[2-(4-amin0-piperidiny1) 76 methyl-S-(l-methy1-1H-ind01y1) OXO-l,6-dihyd1‘O-py1‘imidiny1] ?uorO-benzonitrile 4-[2-(4-amin0-piperidiny1)(1H- 77 indazoly1)methy10x0-1,6- o—pyrimidiny1]?u0r0- benzonitrile 4-[2-((4R, 3 S)amin0-3 -?u0r0- 78 piperidiny1)-5 -(4-meth0xy—pheny1) 1-methy10x0-1,6-dihydr0- pyrimidiny1]?u0r0-benzonitrile 4-[2-((4S, 3R)amin0-3 -?u0r0- 79 piperidiny1)-5 -(4-meth0xy—phenyl)- 1-methy10x0-1,6-dihydr0- diny1]?u0r0-benzonitrile 4- [2-(4-dimethylamino—piperidin y1)methy1-5 -(2-methy1-2H-indazol- 6-y1)0x0-1,6-dihydr0-pyrimidin y1] ?uoro—benzonitrile 4-[2'-dimethy1amin0(4- dimethylamino—piperidiny1) 8 methyl0x0-1,6-dihydr0- [5 ,5']bipyrimidiny1y1] ?u0r0- benzonitrile 4- [2-(4-dimethylamino—piperidin 82 y1)methy1(6-methy1-pyridin-3 - y1)0x0-1,6-dihydr0-pyrimidin y1] ?uoro—benzonitrile 4- [5 -(6-dimethy1amin0-pyridin-3 -y1)- 83 1 -methy1(4-methy1amin0-piperidin- 1-y1)0x0-1,6-dihydr0-pyrimidin y1] ?uoro—benzonitrile 4- [2-(4-dimethylamino—piperidin 84 y1)-5 -(2H-indaz01y1)- 1 1 OXO-l,6-dihyd1‘O-py1‘imidiny1] ?uorO-benzonitrile 4-[2-(4-amin0-piperidiny1)(3 - 85 ?u0r0meth0xy-phenyl) deuteratedmethy10X0- 1 ,6-dihydr0- pyrimidiny1]?u0r0-benzonitrile 4-[2-(4-amin0-piperidiny1)(3 - ?u0r0deuteratedmethoxy—phenyl)- 1-methy10x0-1,6-dihydr0- pyrimidiny1]?u0r0-benzonitrile 2-?u0r0[1-methy1[4- 87 (methylamin0)piperidiny1]—5 -(1- methylindazol-S-y1)0x0pyrimidin- 4-y1]benzonitrile 4-[2-(4-amin0piperidiny1)(1H- 88 indazol-S-y1)methy1 0X0pyrimidiny1] ?uorobenzonitrile 4-[5-(4-amin0phenyl)(4- aminopiperidiny1)- 1 1 0X0pyrimidiny1] ?uorobenzonitrile 4-[2-(4-amin0piperidiny1) methyl[4-(methy1amin0)pheny1] 0X0pyrimidiny1] ?uorobenzonitrile 4-[2-(4-amin0piperidiny1)-5 - [3 - 9 ?u0r0(methylamin0)phenyl] methyl0x0pyrimidiny1] ?uorobenzonitrile 4-[2-[4-(dimethylamin0)piperidin 92 y1]-5 -(6-meth0xypyridin-3 -y1) methyl0x0pyrimidiny1] ?uorobenzonitrile 4-amin0piperidiny1)-5 -(6- 93 ethoxy—S - ?uoropyridin-3 -y1) methyl0x0pyrimidiny1] ?uorobenzonitrile 4-[2-(4-amin0piperidiny1)-5 -(6- 94 ethoxypyridin-3 -y1)methy1 0X0pyrimidiny1] ?uorobenzonitrile 4-[2-(4-amin0piperidiny1)-5 -(4- 95 pheny1)methyl 0X0pyrimidiny1] ?uorobenzonitrile 4-[2-(4-amin0piperidiny1)[4-(2- hydroxyethoxy)pheny1]— 1 -methy1 0X0pyrimidiny1] ?uorobenzonitrile 97 4-[2-(4-amin0piperidiny1)[4-(2- yethoxy)pheny1]— 1 -methy1 oxopyrimidiny1]benz0nitrile 4-[2-(4-amin0piperidiny1)[4-(2- methoxyethoxy)pheny1]— 1 -methy1 0X0pyrimidiny1] ?uorobenzonitrile 4-[2-(4-amin0piperidiny1)[4-(2- hydroxyethy1)phenyl]methy1 0X0pyrimidiny1] ?uorobenzonitrile 4-amin0piperidiny1)-5 - [4- 100 (hydroxymethyl)pheny1]— 1 -methy1 0X0pyrimidiny1] ?uorobenzonitrile 4-[2-(4-amin0piperidiny1)-5 -(4- 101 heny1)methy1 0X0pyrimidiny1] ?uorobenzonitrile 4-[2-(4-amin0piperidiny1)-5 -(3 - 102 ?uoropheny1)methy1 0X0pyrimidiny1] ?uorobenzonitrile 4-[2-(4-amin0piperidiny1)(3 ,5- 103 di?uoropheny1)methy1 0X0pyrimidiny1] ?uorobenzonitrile 4-[2-(4-amin0piperidiny1)(3 ,4- 104 di?uoropheny1)methy1 0X0pyrimidiny1] ?uorobenzonitrile 4-[2-(4-amin0piperidiny1) 105 (4-methylsu1f0ny1pheny1) 0X0pyrimidiny1] ?uorobenzonitrile 4-[2-(4-amin0piperidiny1)-5 -(4- 106 ch10r0pheny1)methy1 0X0pyrimidiny1] ?uorobenzonitrile 4-[2-(4-amin0piperidiny1)-5 - [4- 107 (methoxymethyl)pheny1] methy1 0X0pyrimidiny1] ?uorobenzonitrile 4-[2-(4-amin0piperidiny1) 108 methyl0x0pyrimidiny1] ?uorobenzonitrile 4-amin0-piperidiny1) 109 cyclopropylmethy10X0- 1 ,6-dihydr0- pyrimidiny1]?u0r0-benzonitrile 4-[2-(4-amin0-piperidiny1) 110 cyclopropylmethy10X0- 1 ,6-dihydr0- pyrimidiny1]?u0r0-benzonitrile 2-(4-amin0-piperidiny1)(4- 111 ch10r0-3 -?u0r0-pheny1)-5 -(4- methoxy—pheny1)-3 -methy1-3H- pyrimidin0ne min0-piperidiny1)(4- 112 hydroxy—pheny1)-3 -methy1-5 -(1- methyl-1H-ind01-5 -y1)-3 H-pyrimidin- 4-0ne 2-(4-amin0-piperidiny1)(4- 113 ?uoro—pheny1)-3 -methy1-5 -( 1 -methy1- 1H-ind01-5 -y1)-3H-pyrimidin0ne 2-(4-amin0-piperidiny1)-3 -methy1- 114 5-(1-methy1-1H-ind01y1)pheny1- 3 H-pyrimidin0ne 2-(4-amin0-piperidiny1)-5 -(3 - 115 ?u0r0meth0xy-pheny1)-3 -methy1 pyridiny1-3H-pyrimidin0ne min0-piperidiny1)-3 -methy1- 116 5-(1-methy1-1H-ind01-5 -y1)pyridin- 4-y1-3H-pyrimidin0ne 2-(4-amin0-piperidiny1)(4- 117 methoxy—pheny1)-3 1-5 -(1- methyl-1H-ind01-5 -y1)-3 H-pyrimidin- 4-0ne 3 -[2-(4-amin0piperidiny1)-5 -(3 - 118 ?u0r0meth0xypheny1)methy1 oxopyrimidiny1]benz0nitrile 2-[2-(4-aminopiperidiny1)-5 -(3 - ?uoromethoxypheny1)methy1 oxopyrimidiny1]benzonitri1e 2-(4-amino-piperidiny1)-5 -(3 - ?uoromethoxy—pheny1)methy1 0X0-1,6-dihydro-pyrimidine carbonitrile 2-(4-amino-piperidiny1)(4- cyano-3 -?uoro-pheny1)methy1 oxo-1,6-dihydro-pyrimidine-5 - itrile 4-[2-(4-aminopiperidiny1)-5 -(4- methoxypheny1)oxo-1H-pyrimidin- 4-y1] ?uorobenzonitri1e In some embodiments, the substituted cyclic derivative compound described herein has the structure provided in Table 2.
TABLE 2 Preparation of the Substituted Heterocyclic Derivative Compounds The compounds used in the reactions described herein are made according to organic synthesis techniques known to those skilled in this art, starting from commercially ble chemicals and/or from compounds described in the chemical literature. "Commercially available chemicals" are obtained from standard commercial sources ing Acros Organics (Pittsburgh, PA), Aldrich Chemical (Milwaukee, WI, including Sigma Chemical and Fluka), Apin Chemicals Ltd. (Milton Park, UK), Avocado Research (Lancashire, UK), BDH Inc. to, ), Bionet (Cornwall, UK), Chemservice Inc. (West Chester, PA), Crescent al Co. (Hauppauge, NY), Eastman Organic Chemicals, Eastman Kodak Company (Rochester, NY), Fisher Scienti?c Co.
(Pittsburgh, PA), Fisons als (Leicestershire, UK), Frontier Scienti?c (Logan, UT), ICN Biomedicals, Inc. (Costa Mesa, CA), Key Organics (Cornwall, UK), Lancaster Synthesis (Windham, NH), Maybridge Chemical Co. Ltd. (Cornwall, UK), Parish Chemical Co. (Orem, UT), Pfaltz & Bauer, Inc. (Waterbury, CN), Polyorganix (Houston, TX), Pierce Chemical Co. (Rockford, IL), Riedel de Haen AG (Hanover, Germany), Spectrum Quality Product, Inc. (New Brunswick, NJ), TCI America (Portland, OR), Trans World Chemicals, Inc. (Rockville, MD), and Wako Chemicals USA, Inc. (Richmond, Suitable reference books and treatise that detail the synthesis ofreactants useful in the preparation of compounds described herein, or provide references to articles that describe the ation, e for example, "Synthetic c Chemistry", John Wiley & Sons, Inc., New York; S. R. Sandler et al., "Organic Functional Group Preparations," 2nd Ed., Academic Press, New York, 1983; H. 0. House, "Modern Synthetic Reactions", 2nd Ed., W. A. in, Inc. Menlo Park, Calif 1972; T. L. Gilchrist, "Heterocyclic Chemistry", 2nd Ed., John Wiley & Sons, New York, 1992; J. March, ced Organic Chemistry: Reactions, isms and Structure", 4th Ed., Wiley-Interscience, New York, 1992. Additional suitable reference books and treatise that detail the synthesis of reactants useful in the preparation of compounds described herein, or provide references to articles that describe the preparation, include for example, Fuhrhop, J. and Penzlin G. "Organic Synthesis: Concepts, Methods, Starting als", Second, Revised and Enlarged Edition (1994) John Wiley & Sons ISBN: 329074-5; Hoffman, R.V. "Organic Chemistry, An Intermediate Tex " (1996) Oxford University Press, ISBN 0509618-5; Larock, R. C. ehensive c Transformations: A Guide to onal Group Preparations" 2nd Edition (1999) Wiley-VCH, ISBN: 0- 4714; March, J. "Advanced Organic Chemistry: Reactions, Mechanisms, and Structure" 4th Edition (1992) John Wiley & Sons, ISBN: 060180-2; Otera, J. r) "Modern Carbonyl Chemistry" (2000) Wiley-VCH, ISBN: 329871-1; Patai, S. "Patai's 1992 Guide to the Chemistry of Functional Groups" (1992) Interscience ISBN: 093022-9; Solomons, T. W. G. "Organic Chemistry" 7th Edition (2000) John Wiley & Sons, ISBN: 019095-0; l, J.C., "Intermediate Organic Chemistry" 2nd Edition (1993) Wiley-Interscience, ISBN: 057456-2; "Industrial Organic Chemicals: Starting als and ediates: An Ullmann's Encyclopedia" (1999) John Wiley & Sons, ISBN: 329645-X, in 8 volumes; "Organic Reactions" (1942-2000) John Wiley & Sons, in over 55 s; and "Chemistry of onal Groups" John Wiley & Sons, in 73 volumes.
Speci?c and analogous reactants are optionally identi?ed through the indices of known chemicals prepared by the Chemical Abstract Service ofthe American Chemical Society, which are available in most public and university libraries, as well as through on-line databases (contact the American Chemical Society, Washington, DC for more details). Chemicals that are known but not commercially available in catalogs are optionally prepared by custom chemical synthesis houses, where many ofthe rd chemical supply houses (e.g., those listed above) e custom synthesis services. A reference for the preparation and selection ofpharmaceutical salts ofthe substituted heterocyclic derivative compounds described herein is P. H. Stahl & C. G. Wermuth "Handbook ofPharmaceutical Salts", Verlag Helvetica Chimica Acta, Zurich, 2002.
The substituted heterocyclic derivative nds are prepared by the general synthetic route described below in Scheme 1.
Scheme 1 R2 32 Cl N Cl Cl N Cl Cl N Cl \ N \ HN\ Cl N~ | Y \ | Y R1-X | Y R2' | \Y R2.
, /N _, Nt Cl _. N Cl Cl R1 c. ‘R1 Cl OH 0 o A B c D R \ R2 R2 3 ||3ox H R4\ /OH R3 N N B R N N ‘R I I 3 \ \ ‘R I OH | I 2 OH | I 2 Cl ‘R1 R4 ‘R1 0 O E F Referring to Scheme 1, nd A is selectively hydrolyzed to give compound B.
Compound C is obtained from N-alkylation of compound B with a variety of alkyl halides R1-X. Selective displacement oftrichloride compound C is carried out with a variety of amines HN(R2)(R2’) under basic conditions to form compound D. nd E is prepared from compound D under palladium-mediated cross coupling conditions with boronic acids, e. g. R3-B(OH)2, or boronic esters. Compound F is prepared from compound E under ium-mediated cross coupling conditions with boronic acids, e.g. R3-B(OH)2, or boronic esters.
Pharmaceutical Compositions of the Substituted cyclic Derivative Compounds In certain embodiments, the substituted heterocyclic derivative compound as described herein is administered as a pure chemical. In other embodiments, the substituted heterocyclic derivative nd described herein is combined with a pharmaceutically suitable or acceptable carrier (also referred to herein as a pharmaceutically suitable (or acceptable) excipient, physiologically suitable (or acceptable) excipient, or physiologically suitable (or acceptable) carrier) selected on the basis of a chosen route of stration and standard pharmaceutical practice as bed, for example, in Remington: The Science and Practice ofPharmacy (Gennaro, 21St Ed. Mack Pub. Co., Easton, PA (2005)).
Provided herein is a pharmaceutical composition comprising at least one substituted heterocyclic derivative compound, or a stereoisomer, pharmaceutically able salt, hydrate, solvate, or N-oxide thereof, together with one or more pharmaceutically able carriers. The carrier(s) (or excipient(s)) is acceptable or suitable if the carrier is ible with the other ingredients of the composition and not deleterious to the ent (i. e., the subject) of the composition.
One embodiment provides a pharmaceutical composition comprising a compound ofFormula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically able excipient. One ment provides a pharmaceutical composition comprising a compound of Formula (I a), or a pharmaceutically able salt thereof, and a pharmaceutically acceptable ent. One embodiment provides a pharmaceutical composition comprising a compound of Formula (lb), or a pharmaceutically acceptable salt f, and a pharmaceutically acceptable excipient.
In certain embodiments, the substituted heterocyclic derivative compound as bed by Formula (I) is substantially pure, in that it ns less than about 5%, or less than about 1%, or less than about 0.1%, of other c small molecules, such as ted intermediates or sis by-products that are created, for example, in one or more ofthe steps of a synthesis method.
Suitable oral dosage forms include, for example, tablets, pills, sachets, or capsules of hard or soft gelatin, methylcellulose or of another suitable material easily dissolved in the digestive tract. In some embodiments, suitable nontoxic solid carriers are used which include, for e, ceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, , cellulose, glucose, sucrose, magnesium carbonate, and the like. (See, e. g. The Science and Practice ofPharmacy , Remington: (Gennaro, 21St Ed. Mack Pub. Co., Easton, PA (2005)).
The dose ofthe composition comprising at least one substituted heterocyclic tive compound as described herein differ, depending upon the patient's (e.g, human) condition, that is, stage of the disease, general health status, age, and other factors.
Pharmaceutical compositions are administered in a manner appropriate to the disease to be treated (or prevented). An appropriate dose and a suitable duration and frequency of administration will be determined by such factors as the condition of the patient, the type and severity of the patient's disease, the particular form of the active ingredient, and the method of administration. In general, an appropriate dose and treatment regimen provides the composition(s) in an amount sufficient to provide therapeutic and/or prophylactic benefit (e.g, an improved clinical outcome, such as more frequent te or partial remissions, or longer disease-free and/or overall survival, or a lessening of symptom severity. Optimal doses are generally determined using mental models and/or clinical trials. The optimal dose s upon the body mass, weight, or blood volume ofthe patient.
Oral doses typically range from about 1.0 mg to about 1000 mg, one to four times, or more, per day.
Use of the Substituted cyclic Derivative Compounds Epigenetics is the study of heritable changes in gene expression caused by mechanisms other than the ying DNA sequence. Molecular isms that play a role in epigenetic regulation include DNA methylation and chromatin/histone modifications.
The genomes of eukaryotic sms are highly organized within the nucleus ofthe cell. Tremendous compaction is required to package the 3 billion tides of the human genome into the nucleus of a cell. Chromatin is the complex ofDNA and protein that makes up chromosomes. Histones are the major protein component of chromatin, acting as spools around which DNA winds. Changes in tin ure are ed by covalent modi?cations of histone proteins and by non-histone binding proteins. Several classes of enzymes are known which modify histones at various sites.
There are a total of six classes of histones (HI, H2A, H2B, H3, H4, and H5) organized into two groups: core histones (H2A, H2B, H3, and H4) and linker histones (HI and H5). The basic unit of chromatin is the nucleosome, which consists of about 147 base pairs ofDNA wrapped around the core histone octamer, consisting oftwo copies each ofthe core histones H2A, H2B, H3, and H4.
Basic nucleosome units are then ?arther organized and condensed by the aggregation and folding ofnucleosomes to form a highly condensed chromatin structure.
A range of different states of condensation are possible, and the tightness of chromatin structure varies during the cell cycle, being most compact during the process of cell division.
Chromatin structure plays a critical role in ting gene transcription, which cannot occur ently from highly condensed chromatin. The chromatin structure is controlled by a series of post translational modifications to histone proteins, notably histones H3 and H4, and most commonly within the histone tails which extend beyond the core nucleosome structure. These modifications are acetylation, methylation, orylation, ribosylation sumoylation, ubiquitination, citrullination, deimination, and biotinylation. The core of es H2A and H3 can also be modi?ed. Histone modifications are integral to diverse biological ses such as gene regulation, DNA repair, and chromosome condensation.
Histone methylation is one of the most important tin marks; these play important roles in transcriptional regulation, DNA-damage response, heterochromatin formation and maintenance, and X-chromosome inactivation. A recent discovery also revealed that histone methylation affects the splicing outcome of pre- mRNA by in?uencing the recruitment of splicing regulators. Histone methylation includes mono-, di-, and tri-methylation of lysines, and mono-, symmetric di-, and asymmetric di-methylation of arginines. These modi?cations can be either an ting or repressing mark, depending on the site and degree of methylation.
Histone Demethylases A "demethylase" or "protein demethylase," as referred to herein, refers to an enzyme that removes at least one methyl group from ptide. Demethylases comprise a ijC domain, and can be a methyl-lysine or methyl-arginine demethylase.
Some demethylases act on histones, e. g., act as a histone H3 or H4 demethylase. For example, an H3 demethylase may demethylate one or more ofH3K4, H3K9, H3K27, H3K36 and/or H3K79. Altemately, an H4 demethylase may demethylate e H4K20.
Demethylases are known which can demethylate either a mono-, di- and/or a tri- methylated substrate. Further, histone demethylases can act on a methylated core histone substrate, a mononucleosome substrate, a dinucleosome substrate and/or an oligonucleosome substrate, peptide substrate and/or chromatin (e.g., in a cell-based assay).
The ?rst lysine demethylase discovered was lysine speci?c demethylase l (LSDl/KDMl), which demethylates both mono- and di-methylated H3K4 or H3K9, using ?avin as a cofactor. A second class of Jumonji C (ijC) domain containing histone demthylases were predicted, and con?rmed when a H3K36 demethylase was found used a formaldehyde release assay, which was named ijC domain containing histone demethylase l (JHDMl/KDMZA).
More ijC domain-containing proteins were subsequently identi?ed and they can be phylogenetically clustered into seven ilies: JHDMl, JHDM2, JHDM3, JMJDZ, JARID, PHF2/PHF8, Y, and ijC domain only.
LSD-I -speci?c demethylase 1 (LSDl) is a histone lysine demethylase that speci?cally demethylates thylated and ylated histone H3 at K4 and also demethylates dimethylated histone H3 at K9. Although the main target of LSDl appears to be mono- and hylated histone lysines, speci?cally H3K4 and H3K9, there is eVidence in the literature that LSD 1 can demethylate methylated lysines on stone proteins like p53, E2Fl Dnmtl and STAT3.
LSD 1 has a fair degree of structural similarity and amino acid ty/homology to polyamine oxidases and monoamine oxidases, all ofwhich (i. e., MAO-A, MAO-B and LSDl) are ?avin dependent amine oxidases which catalyze the oxidation of nitrogen-hydrogen bonds and/or en-carbon bonds. LSDl also es an N—terminal SWRIM domain. There are two transcript variants ofLSDl produced by alternative splicing.
In some embodiments, the compounds disclosed herein are capable of inhibiting LSDl activity in a ical sample by contacting the biological sample with a substituted heterocyclic nd as disclosed herein. In some embodiments, a substituted cyclic compound as disclosed herein is capable of modulating the level of histone 4 lysine 3 methylation in the biological sample. In some embodiments, a substituted heterocyclic compound as sed herein is capable of modulating histone- 3 lysine-9 methylation levels in the biological sample.
The substituted heterocyclic compounds disclosed herein lack signi?cant MAO-A or MAO-B inhibitory activity. In some embodiments, a tuted heterocyclic nd as disclosed herein inhibits LSDl inhibitory activity to a greater extent than MAO-A and/or MAO-B inhibitory activity.
One embodiment provides a method ofregulating gene transcription in a cell comprising inhibiting lysine-specific demethylase 1 activity by exposing the lysinespecif1c demethylase l enzyme to a compound of Formula (I). One embodiment provides a method ofregulating gene transcription in a cell comprising inhibiting lysine-specific demethylase 1 activity by exposing the lysine-specific demethylase l enzyme to a compound ofFormula (Ia). One embodiment provides a method ofregulating gene transcription in a cell comprising inhibiting lysine-specific demethylase 1 activity by exposing the lysine-specific demethylase l enzyme to a compound of Formula (Ib).
Methods of Treatment ] Disclosed herein are methods of modulating demethylation in a cell or in a subject, either generally or with respect to one or more c target genes.
Demethylation is modulated to control a variety of ar ?anctions, ing without limitation: differentiation; proliferation; apoptosis; tumorigenesis, leukemogenesis or other oncogenic transformation events; hair loss; or sexual differentiation.
] One embodiment provides a method ting cancer in a patient in need thereof, comprising administering to the patient a compound of Formula (I), or a pharmaceutically acceptable salt thereof One ment provides a method of treating cancer in a patient in need thereof, comprising stering to the patient a compound of Formula (Ia), or a pharmaceutically acceptable salt thereof One embodiment provides a method oftreating cancer in a patient in need thereof, comprising administering to the patient a compound of Formula (lb), or a pharmaceutically able salt thereof.
In a farther embodiment is the method for treating cancer in a subject wherein the cancer is selected from prostate cancer, breast cancer, r , lung cancer or ma. In a farther embodiment is the method for treating cancer in a t wherein the cancer is selected from acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), small cell lung cancer (SCLC), non-small cell lung cancer ), neuroblastoma, small round blue cell tumors, or glioblastoma.
Other embodiments and uses will be apparent to one skilled in the art in light ofthe present disclosures. The following examples are provided merely as illustrative of s embodiments and shall not be construed to limit the invention in any way.
EXAMPLES 1. Chemical Synthesis Unless otherwise noted, reagents and solvents were used as received from commercial suppliers. Anhydrous solvents and oven-dried glassware were used for synthetic transformations sensitive to moisture and/or oxygen. Yields were not optimized. Reaction times are approximate and were not optimized. Column chromatography and thin layer chromatography (TLC) were performed on silica gel unless otherwise noted. Spectra are given in ppm (8) and coupling constants, J are reported in Hertz. For proton spectra the solvent peak was used as the reference peak.
Preparation 1A: 2,5,6-trichloropyrimidinol CI NYCI To a solution of 6-tetrachloropyrimidine (5 g, 22.9 mmol) in THF (50 mL) was added lN NaOH (31 mL, 31.2 mmol) dropwise, and the mixture was stirred overnight at RT. The solution was acidi?ed with lN HCl and extracted with DCM (3x).
The cs were combined, dried, and concentrated in vacuo. The solids were slurried in EtZO for 30 min at RT, ?ltered, washed with EtzO, and dried to give 3.0 g (66%) of the title compound. [M+H] Calc’d for C4HC13N20, 201; Found, 201.
Preparation 1B: 2,5,6-trichloromethylhydropyrimidinone To a mixture of 2,5,6-trichloropyrimidinol (1 g, 5.0 mmol) and K2C03 (759 mg, 5.5 mmol) in THF (50 mL) at 0 0C was added iodomethane (714 mg, 5.0 mmol) dropwise, and the reaction was stirred at RT overnight. The reaction mixture was diluted with ethyl acetate (EA). The organic phase was washed with brine, dried and concentrated in vacuo. The residue was puri?ed by silica gel chromatography (10:1, PE:EA) to give 760 mg (71%) ofthe title compound. 1H NMR (400 MHZ, CDC13)I 8 3.74 (s, 3 H). [M+H] Calc’d for C5H3C13N20, 213; Found, 213. ation 1C: N—[1-(5,6-dichloromethyloxo(3-hydropyrimidinyl))(4- piperidyl)](tert-butoxy)carboxamide ||30c CI N N A solution of 2,5,6-trichloromethylhydropyrimidinone (426 mg, 2.0 mmol), DIEA (536 mg, 4.0 mmol) and tert-butyl piperidinylcarbamate (400 mg, 2 mmol) in DMF (10 mL) was heated at 120 0C for 1 h. The solvent was removed in vacuo and the residue was puri?ed by silica gel chromatography (1 :1, PE:EA) to give 550 mg (73%) ofthe title compound. 1H NMR (400 MHz, CDC13)C 8 1.45 (s, 9H),1.50-1.58 (m, 2H), .10 (m, 2H), 2.98-3.05 (m, 2H), 3.48 (s, 3 H), .56 (m, 2H), 3.70 (s, 1H), 4.52 (s, 1H). [M+H] Calc’d for C15H22C12N403, 213; Found, 213. ation 1D: utyl 1-(5-chloro(4-cyanophenyl)methyloxo-1,6- dihydropyrimidinyl)piperidinylcarbamate N NO/NH Cl \ [001 1 1] A mixture ofN—[ 1 -(5,6-dichloromethyloxo(3-hydropyrimidinyl))(4- piperidyl)](tert—butoxy)carboxamide (500 mg, 1.3 mmol), 4~cyanophenylboronic acid (195 mg, 1.3 mmol), [l,l’-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (81 mg, 0.13 mmol) and K2C03 (359 mg, 2.6 mmol) in DMF (10 mL) was ?ushed with nitrogen and stirred at 85 0C for 2 h. Water was added, and the mixture was extracted with EA (3x). The organics were combined, washed with water, washed with brine, dried and concentrated in vacuo. The residue was puri?ed puri?ed by silica chromatography (1:1, EA:PE) to give 250 mg (40%) ofthe title compound. 1H NMR (400 MHz, CDC13)I 8 1.45 (s, 9H), 1.54-1.61 (m, 2H), 2.05-2.10 (m, 2H), 2.99-3.05 (m, 2H), 3.48-3.56 (s, 5H), 3.70 (s, 1H), 4.56 (s, 1H), 7.73 (d, J: 8.0 Hz, 2H), 7.93 (d, J: 8.0 Hz, 2H). [M+H] Calc’d for C22H26C1N503, 444; Found, 444.
Preparation 1E: tert—butyl l-(4-(4-cyanophenyl)-l-methyloxop-tolyl-l,6- opyrimidinyl)piperidinylcarbamate A mixture of tert—butyl hloro(4-cyanophenyl)-l-methyloxo-l,6- dihydropyrimidinyl)piperidinylcarbamate (200 mg, 0.45 mmol), p-tolylboronic acid (123 mg, 0.90 mmol), [1,1’-bis(di-tert- butylphosphino)ferrocene]dichloropalladium(II) (28 mg, 0.045mol) and K2C03 (124 mg, 0.90 mmol) in DMF (10 mL) was ?ushed with nitrogen and stirred at 85 0C for 2 h.
Water was added, and the mixture was ted with EA (3x). The cs were combined, washed with water, washed with brine, dried and concentrated in vacuo. The residue was puri?ed by silica chromatography (1:1, EA:PE) to give 50 mg (22%) ofthe title compound. [M+H] Calc’d for C29H33N503, 500; Found, 500.
Example 1: 4-(2-(4-aminopiperidin- l -yl)- l -methyloxop-tolyl- l ,6- dihydropyrimidinyl)benzonitrile, HC1 salt 0 NH2 To a solution of utyl l-(4-(4-cyanophenyl)-l-methyloxop-tolyl- 1,6-dihydro pyrimidinyl)piperidinylcarbamate (50 mg, 0.1 mmol) in EA (10 mL) was added a 4N HC1 solution in EA (5 mL) and the mixture was stirred at RT for 2 h.
The solvent was concentrated in vacuo, and the residue was puri?ed by preparative HPLC to give 20 mg (46%) ofthe title nd as the hydrochloride salt. 1H NMR (400 MHz, CDC13): 8 1.74-1.79 (m, 2H), 2.00-2.04 (m, 2H), 2.21 (s, 3H), 2.96-3.03 (m, 2H), 3.29-3.03 (m, 1H), 3.48 (s, 3H), 3.71-3.74 (m, 2H), 6.89 (d, J: 8.0 Hz, 2H), 6.99 (d, J: 8.0 Hz, 2 H), 7.38 (d, J: 8.0 Hz, 2H), 7.44 (d, J: 8.4 Hz, 2H). [M+H] Calc’d for C24H25N50, 400; Found, 400.
Example 2: 4-[2-(4-amino-piperidinyl)(4-methoxy-phenyl)methyloxo-1 ,6- dihydro-pyrimidinyl]-benzonitrile NY.CVNH The title compound was prepared as the hydrochloride salt in 5% overall yield according to the general procedure for the preparation of e 1. 1H NMR (400 MHz, CDgOD): 8 .78 (m, 2H), 2.00-2.03 (m, 2H), 2.98-3.02 (m, 2H), .00 (m, 1H), 3.48 (s, 3H), 3.69 (s, 3H), 3.70-3.73 (m, 2H), 6.72 (d, J: 8.8 Hz, 2H), 6.93 (d, J = 8.4 Hz, 2H), 7.39 (d, J: 8.0 Hz, 2H), 7.46 (d, J: 8.0 Hz, 2H). [M+H] Calc’d for C24H25N502, 416; Found, 416.
Example 3: 4-[2-(4-amino-piperidinyl)(6-methoxy-pyridinyl)methyloxo- 1 ,6-dihydro-pyrimidinyl] -benzonitrile The title compound was prepared as the hydrochloride salt in 11% overall yield according to the general ure for the preparation of Example 1. 1H NMR (400 MHz, CDgOD): 8 1.87-1.95 (m, 2H), 2.14-2.17 (m, 2H), .24 (m, 2H), 3.43-3.48 (m, 1H), 3.62 (s, 3H), 3.93-3.98 (m, 2H), 4.23 (s, 3H), 7.46 (d, J: 9.2 Hz, 1H), 7.63 (d, J = 8.0 Hz, 2H), 7.71 (d, J: 8.4 Hz, 2H), 8.12 (dd, J: 8.8, 1.6 Hz, 1H), 8.28 (d, J: 2.0 Hz, 1H). [M+H] Calc’d for C23H24N602, 417; Found, 417.
Example 4: 4-[2-(4-amino-piperidinyl)methyl(6-methyl-pyridinyl)oxo- 1 ,6-dihydro-pyrimidinyl] -benzonitrile NC NH2 N\ N\ I/ 0 The title compound was prepared as the hydrochloride salt in 4% overall yield according to the general procedure for the preparation of Example 1. 1H NMR (400 MHz, CD30D)C 5 1.79-1.80 (m, 2H), .05 (m, 2H), 2.66 (s, 3H), 3.04-3.09 (m, 2H), 3.30-3.34 (m, 1H), 3.50 (s, 3H), 3.83-3.88 (m, 2H), 7.48 (d, J: 8.4 Hz, 2H), 7.58 (d, J: 8.4 Hz, 2H), 7.64 (d, J: 8.4 Hz, 1H), 8.00 (dd, J: 8.4, 2.0 Hz, 1H), 8.54 (d, J: 8.0 Hz, 1H). [M+H] Calc’d for C23H24N6O, 401; Found, 401.
Example 5: 4-[2-(4-amino-piperidinyl)(4-methoxy-phenyl)methyloxo-1 ,6- dihydro-pyrimidinyl]-benzonitrile 0 NONH2 The title compound was prepared as the hydrochloride salt in 7% overall yield according to the general procedure for the preparation of e 1. 1H NMR (400 MHz, CDgOD): 8 1.89-1.95 (m, 2H), 2.15-2.18 (m, 2H), 3.14-3.18 (m, 2H), 3.44-3.46 (m, 1H), 3.60 (s, 3H), 3.88-3.90 (m, 5H), 6.79 (d, J: 8.4 Hz, 1H), 6.96-7.02 (m, 2H), 7.54 (d, J: 8.0 Hz, 2H), 7.64 (d, J: 8.0 Hz, 2H). [M+H] Calc’d for C24H24FN502, 434; Found, 434.
Example 6: 4-[2-(4-amino-piperidinyl)(4-methoxy-phenyl)methyloxo-1 ,6- o-pyrimidinyl]?uoro-benzonitrile The title compound was prepared as the hydrochloride salt in 5% overall yield ing to the general procedure for the preparation of Example 1. 1H NMR (400 MHz, CDgOD): 8 1.83-1.89 (m, 2H), 2.10-2.13 (m, 2H), 3.05-3.11 (m, 2H), 3.35-3.38 (m, 1H), 3.55 (s, 3H), 3.76 (s, 3H), 3.77-3.82 (m, 2H), 6.84 (d, J: 8.8 Hz, 2H), 7.04 (d, J = 8.8 Hz, 2H), 7.21 (d, J: 8.0 Hz, 1H), 7.35 (d, J: 8.0 Hz, 1H) v 7.53-7.56 (m, 1H).
[M+H] Ca1c’d for FN502, 434; Found, 434.
Preparation 7A: tert—butyl hloro(3-?uorocyanopheny1)methy1oxo-1,6- dihydropyrimidin—2-y1)piperidiny1carbamate F §oc NC NH CI \ A mixture ofN—[1-(5,6-dichloromethy1oxo(3-hydropyrimidiny1))(4- piperidy1)](tert—butoxy)carboxamide (150 g, 0.40 mol), 3-?uorocyanopheny1boronic acid (65.8 g, 0.40 mol), Pd(Ph3P)4 (9.3 g, 8 mmol) and 0.4 N N32C03 (2 L, 0.80 mol) in ACN (4 L) was ?ushed with nitrogen and d at 85 0C for 2 h. Water was added and the mixture was extracted with EA (3x). The organics were combined, washed with water, washed with brine, dried and concentrated in vacuo. The residue was puri?ed puri?ed by silica chromatography (1 :1, EA:PE) to give 95 g (57%) of the title compound. 1H NMR (400 MHz, CDC13): 8 1.45 (s, 9 H),1.54-1.61 (m, 2H), .13 (m, 2H), 2.99-3.08 (m, 2H), 3.53-3.58 (s, 5H), 3.70 (s, 1H), 4.54 (d, J: 6.0 Hz, 1H), 7.68-7.80 (m, 3 H).
Preparation 7B: tert—butyl N—[1-[4-(4-cyano?uoropheny1)(3-?uoro methoxypheny1)methy1oxopyrimidiny1]piperidiny1] carbamate A mixture of (tert—butoxy)-N—{1-[5-chloro(4-cyano?uoropheny1)—3- methyloxo(3-hydropyrimidiny1)](4-piperidy1)}carboxamide (1 g, 2.169 mol), 3- ?uoromethoxy benzeneboronic acid (740 mg, 4.338 mmol), Pd(dppf)C12 (480 mg, 0.651 mmol) and N32C03 (690 mg, 6.51 mmol) in dioxane:HzO (3:1, 15 mL) was ?ushed with nitrogen, capped and stirred at 145 0C for 2 h in the microwave. The reaction mixture was trated and the residue was puri?ed by PC (1 :1, EA:PE) to give 800 mg (71%) ofthe title compound. [M+H] Ca1c’d for C29H31F2N504, 552; Found, 552. 1H NMR (400 MHz,CDC13): 8 ppm 1.46 (s, 9 H), 1.60 (d, J=10.11 Hz, 2 H), 2.11 (d, J=11.62 Hz, 2 H), 3.06 (t, J=12.00 Hz, 2 H), 3.54 (s, 3 H), 3.60 (d, J=13.64 Hz, 2 H), 3.72 (br. s., 1 H), 3.88 (s, 3 H), 4.52 (br. s., 1 H), 6.79 - 6.89 (m, 2 H), 6.97 (d, J=12.38 Hz, 1 H), 7.13 (d, J=8.34 Hz, 1 H), 7.31 (d, J=9.85 Hz, 1 H), 7.42 (br. s., 1 H).
Example 7: 4-[2-(4-amino-piperidinyl)-5 oromethoxy-phenyl)methyl oxo- 1 ,6-dihydro-pyrimidinyl] ?uoro -benzonitrile To a solution of tert—butyl N-[l-[4-(4-cyano?uorophenyl)(3-?uoro yphenyl)methyloxopyrimidinyl]piperidinyl]carbamate (5.2 g, 9.44 mmol) in EA (20 mL) was added a 1N HCl in EA (30 mL). The e was stirred at RT for 2 h. The solvent was concentrated in vacuo to give the title product as the HC1 salt (4.05 g, 88%). 1H NMR (400 MHz, CD30D)C 8 1.77-1.79 (m, 2H), 2.02-2.04 (m, 2H), 2.99-3.04 (m, 2H), 3.26-3.00 (m, 1H), 3.38 (s, 3H), 3.73 (s, 3H), 3.73-3.75 (m, 2H), 6.67-6.68 (m, 1H), 6.84-6.95 (m, 2 H), 7.12-7.14 (m, 1H), 7.24-7.36 (m, 1H) 7.46-7.50 (m, 1H). [M+H] Calc’d for C24H23F2N502, 452; Found, 452.
Example 8: 4-[2-(4-amino-piperidinyl)(6-methoxy-pyridinyl)methyloxo- 1 ,6-dihydro-pyrimidinyl] ?uoro-benzonitrile The title compound was prepared as the hydrochloride salt in 6% l yield ing to the general ure for the preparation of Example 1. 1H NMR (400 MHz, CD30D)I 8 1.79-1.83 (m, 2H), 2.02-2.06 (m, 2H), 3.04-3.11 (m, 2H), 3.21-3.22 (m, 1H), 3.49(s, 3H), 3.81-3.85 (m, 2H), 4.12 (s, 3H), 7.22-7.24 (m, 1H), 7.38 (d, J: 9.2 Hz, 1H), 7.49 (d, J: 9.2 Hz, 1H), 7.57-7.61 (m, 1H), 8.04-8.07 (m, 1H), 8.21 (s, 1H).
[M+H] Calc’d for C23H23FN602, 435; Found, 435.
Example 9: 4-[2-(4-amino-piperidinyl)(6-methoxy-pyridinyl)methyloxo- 1 ,6-dihydro-pyrimidinyl] ?uoro-benzonitrile NC NH2 u\ N\ l/ 0 The title compound was prepared as the hydrochloride salt in 8% overall yield according to the general procedure for the preparation of Example 1. 1H NMR (400 MHz, CDgOD): 8 1.92-1.96 (m, 2H), 2.16-2.19 (m, 2H), 2.80 (s, 3H), 3.19-3.25 (m, 2H), 3.45-3.49 (m, 1H), 3.62 (s, 3H), 3.96-3.99 (m, 2H), 7.34 (d, J: 8.0 Hz, 1H), 7.60 (d, J: 7.2 Hz, 1H), 7.71 (t, J: 7.6 Hz, 1H), 7.80 (d, J: 8.4 Hz, 1H), 8.18 (d, J: 8.4 Hz, 1H), 8.71 (s, 1H). [M+H] Calc’d for C23H23FN6O, 419; Found, 419.
Example 10: 4-amino-piperidinyl)(6-ethyl-pyridinyl)methyloxo-1,6- dihydro-pyrimidinyl]-benzonitrile The title compound was prepared as the hydrochloride salt in 7% overall yield according to the general procedure for the preparation of e 1. 1H NMR (400 MHz, CD30D)C 8 1.30 (t, J: 4.0 Hz, 3H), 1.83-1.88 (m, 2H), 2.06-2.09 (m, 2H), 2.96- 2.99 (m, 2H), 3.09-3.16 (m, 2H), 3.26-3.31 (m, 1H), 3.51 (s, 3H), 3.86-3.89 (m, 2H), 7.35 (d, J: 8.0 Hz, 2H), 7.61 (d, J: 8.0 Hz, 2H), 7.71 (d, J: 8.4 Hz, 1H), 8.08 (d, J: 8.4 Hz, 1H), 8.57 (s, 1H). [M+H] Calc’d for C24H26N6O, 415; Found, 415.
Example 1 1: 2-?uoro[5-(4-methoxy-phenyl)methyl(4-methylamino-piperidin- 1-yl)oxo- 1 ,6-dihydro-pyrimidinyl] -benzonitrile The title compound was prepared as the hydrochloride salt in 7% overall yield according to the l ure for the preparation of Example 1. 1H NMR (400 MHz, cogoo); 5 .90 (m, 2H), 2.19—2.23 (m, 2H), 2.75 (s, 3H) 6.06-3.12 (m, 2H), 3.32-3.36 (m, 1H), 3.56 (s, 3H), 3.76 (s, 3H), 3.84-3.87 (m, 2H), 6.84 (d, .1: 8.4 Hz, 2H), 7.04 (d, J: 8.4 Hz, 2H), 7.22 (d, J: 8.0 Hz, 1H), 7.36 (d, J: 10.8 Hz, 1H), 8.54-7.58 (m, 1H). [M+H] Calc’d for C25H26FN502, 448; Found, 448.
Example 12: 2-?uoro[5-(3-?uoromethoxy-phenyl)methyl(4-methylamino- piperidinyl)oxo- 1 ,6-dihydro -pyrimidinyl] -benzonitrile The title compound was prepared as the hydrochloride salt in 7% overall yield ing to the general procedure for the preparation of Example 1. 1H NMR (400 MHz, CDgOD): 8 1.78-1.88 (m, 2H), .20 (m, 2H), 2.73 (s, 3H) v3.05-3.11 (m, 2H), 3.30-3.35 (m, 1H), 3.54 (s, 3H), 3.82 (s, 3H), 3.83-3.86 (m, 2H), 6.76 (d, J: 8.4 Hz, 1H), .99 (m, 2H), 7.20 (d, J: 8.4 Hz, 1H), 7.38 (d, J: 10.4 Hz, 1H), 8.55- 7.589 (m, 1H). [M+H] Calc’d for C25H25F2N502, 466; Found, 466.
Preparation 13A: 2,6-dichloroethyl-3H-pyrimidinone CI N | A solution of2,6-dichloro-pyrimidinol (1.0 g, 6.1 mmol) and K2C03 (1.1 g, 7.9 mmol) in DMF (10 mL) was stirred at RT for 15 min. The on mixture was cooled to 0 0C, and iodoethane (1.1 mL, 6.7 mrnol) was added dropwise. After stirring overnight at RT, the reaction mixture was diluted with EA, washed with brine, dried (Na2S04) and concentrated in vacuo. The residue was puri?ed by silica chromatography (20:1, EA:PE) to give 330 mg (28%) ofthe title compound. 1H NMR (400 MHZ, CDC13): 8 1.37 (t, J: 7.6 Hz, 3H), 4.76 (q, J: 6.8 Hz, 2H), 6.67 (s, 1H). [M+H] Calc’d for C6H6C12N20, 193, 195, 197; Found, 193, 195, 197.
Preparation 13B: [1-(4-chloroethyloxo - 1 ,6-dihydro idiny1)-piperidin yl]-carbamic acid tert—butyl ester ] A solution of 2,6-dichloroethyl-3H-pyrimidinone (320 mg, 1.64 mmol), DIEA (423 mg, 3.28 mmol) and (tert-butoxy)-N-(4-piperidyl)carboxamide (328 mg, 1.64 mmol) in DMF (10 mL) was heated to 120 0C for 1 h. The solvent was concentrated in vacuo and the residue was puri?ed by silica tography (1 :5, EA:PE) to give 210 mg (36%) ofthe title compound as a yellow solid. 1H NMR (400 MHz, CDC13)C 8 1.25- 1.32 (m 2H), 1.35 (t, J: 7.2 Hz, 3H), 1.96-2.02 (m, 2H), 2.98-3.06 (m, 2H), 3.70 (br, 1H), 4.30 (q, .1: 5.2 Hz, 2H), 4.44 (br, 1H), 4.57-4.61 (m, 2H), 5.95 (s, 1H). [M+H] Calc’d for C16H25C1N403, 357, 359; Found, 357, 359.
Preparation 13C: {1-[4-(4-cyano?uoro-phenyl)ethyloxo-1,6-dihydro- pyrimidinyl]-piperidinyl} -carbamic acid tert—butyl ester F $00 N NH A mixture of [1-(4-chloroethyloxo-1,6-dihydro-pyrimidinyl)-piperidin- carbamic acid tert—butyl ester (210 mg, 0.59 mrnol) in CH3CN (10 mL), 3-?uoro cyanophenylboronic acid (126 mg, 0.77 mmol), Pd(PPh)4 (14 mg, 0.012 mmol) and 0.4 M N32C03 (4.5 mL, 1.77 mrnol) was stirred at 90 0C overnight under N2 atmosphere.
The organic was concentrated in vacuo, and the aqueous extracted with DCM (2x). The combined organics were washed with brine, dried (Na2S04) and concentrated. The residue was d by silica chromatography (1 :2, EA:PE) to give 185 mg (64%) ofthe title compound as a yellow solid. [M+H] Calc’d for C23H28FN503, 442; Found, 442.
Example 13: 4-[2-(4-amino-piperidinyl)ethyloxo-1,6-dihydro-pyrimidinyl]- 2-?uoro-benzonitrile \ NH2 To a mixture of {1-[4-(4-cyano?uoro-phenyl)ethyloxo-1,6-dihydro- pyrimidinyl]-piperidinyl} -carbamic acid tert-butyl ester (180 mg, 0.41 mmol) in EA (5 mL) was added a 4 M solution of HCl in EA (3 mL). The reaction e was stirred for 30 min. The solvent was evaporated in vacuo to give 150 mg of the titled compound (97 %) as a yellow solid (HCl salt). 1H NMR (400 MHz, C 8 1.28 (t, J = 7.2 Hz, 1H), 1.48-1.52 (m, 2H), 1.99-2.02 (m, 2H), 2.94-3.01 (m, 2H), 3.33-3.38 (m, 1H), 6.81 (q, J: 6.8 Hz, 2H), 4.85-4.88 (m, 2 H), 6.95 (s, 1H), 7.73 (t, J: 8.0 Hz, 1H), 7.90-7.95 (m, 2H). [M+H] Calc’d for C18H20FN50, 342; Found, 342.
Preparation 14A: { 1 -[4-(4-cyano -3 -?uoro -phenyl)-5 -cyclopentylethynylmethyl oxo-1,6-dihydro-pyrimidinyl]-piperidinyl}-carbamic acid tert—butyl ester A e of tert—butyl 1-(5-chloro(3-?uorocyanophenyl)methyl oxo-1,6-dihydropyrimidinyl)piperidinylcarbamate (200 mg, 0.43 mmol), ethynyl- cyclopentane (82 mg, 0.87 mrnol), Pd(MeCN)2C12 (4.5mg, 0.017mmol), X-Phos (10 mg, 0.022 mmol) and K2C03 (120 mg, 0.87 mmol) in ACN (15 mL) was stirred overnight at 95 0C in a sealed tube. The reaction mixture was cooled to RT and the solvent was concentrated in vacuo. The residue was puri?ed by silica chromatography (1 :2, EA:PE) to give 100 mg (45%) ofthe title compound. [M+H] Calc’d for FN503, 519; Found, 519.
Example 14: 4-[2-(4-amino-piperidinyl)cyclopentylethynylmethyloxo-1,6- dihydro-pyrimidinyl]?uoro-benzonitrile The title compound was ed as the hydrochloride salt in 70% overall yield ing to the general procedure for the preparation of Example 1. 1H NMR (400 MHz, CDClg): 8 1.50-1.74 (m, 8H), 1.94-1.99 (m, 4H), 2.88-3.01 (m, 4H), 3.51 (s, 3H), 3.60 (d, J: 13.2 Hz, 2H), 7.63-7.67 (m, 1H), 8.07-8.11 (m, 2H). [M+H] Calc’d for C24H26FN50, 419; Found, 419.
Preparation 15A: -trichlorooxo-6H-pyrimidinyl)-acetic acid methyl ester To a solution of 2,5,6-trichloro-3H-pyrimidinone (20.0 g, 0.1 mol) in DMF (150 mL) was added NaH (60 % in mineral oil, 6.0 g, 0.12 mol) in portions at 0 0C and the mixture was stirred for 30 min. cetic acid methyl ester (18.3 g, 0.12 mol) was then added, and the reaction mixture was stirred at RT overnight. The solution was diluted with water (800 mL) and extracted with EA (200 mL, 3x). The combined organics were washed with water (800 mL, 3x), washed with brine (500 mL), dried (Na2S04) and concentrated. The residue was puri?ed by silica chromatography (1 :50, EA:PE) to give 6.0 g ofthe title product (22%). 1H NMR (400 MHZ, CDC13)I 5 3.80 (s, 3H), 5.04 (s, 2H). [M+H] Calc’d for C7H5C13N203, 271; Found, 271.
Preparation 15B: [2-(4-tert—butoxycarbonylamino-piperidinyl)-4,5-dichlorooxo- imidinyl]-acetic acid methyl ester Cl NO To a solution of (2,4,5-trichlorooxo-6H-pyrimidinyl)-acetic acid methyl ester (6.0 g, 22.4 mmol) and piperidinyl-carbamic acid tert—butyl ester (4.9 g, 24.4 mmol) in DMF (50 mL) was added DIPEA (5.7 g, 44.3 mmol) dropwise at RT, and the mixture was stirred overnight. The reaction mixture was diluted with water (500 mL), and the solids were ted by ?ltration. The solids were then dissolved in DCM (100 mL), washed with water (100 mL, 3x), washed with brine (100 mL), dried (Na2S04) and concentrated. The residue was puri?ed by silica chromatography (1 :2 to 1:1, DCM:PE) to give 6.3 g ofthe title product (64%). 1H NMR (400 MHz, CDClg): 5 1.22-1.34 (m, 2H), 1.45 (s, 9H), 1.97-2.03 (m, 2H), .09 (m, 2H), 3.68-3.69 (m, 1H), 3.75 (s, 3H), 4.42-4.44 (m, 3H), 4.84 (s, 2H). [M+H] Calc’d for C17H24C12N405, 435; Found, 435.
Preparation 15C: [2-(4-tert—butoxycarbonylamino-piperidinyl)chloro(4-cyano- 3-?uoro-phenyl)oxo-6H-pyrimidinyl]-acetic acid methyl ester F Y \ NH A e of [2-(4-tert-butoxycarbonylamino-piperidin-l-yl)-4,5-dichloro oxo-6H-pyrimidinyl]-acetic acid methyl ester (5.76 g, 13.2 mmol), 4-cyano?uoro benzeneboronic acid (2.24 g, 16.1 mmol), Pd(PPh3)4 (306 mmol, 0.26 mmol) and N32C03 (2.8 g, 26.5 mrnol) in DMF:H20 (50 mL: 10 mL) was stirred at 65 0C overnight under nitrogen atmosphere. The reaction mixture was concentrated, and the residue was puri?ed by silica chromatography (1:20 to 1:0, EA:PE) to give 2.4 g of the title product (43%). 1H NMR (400 MHz, CDC13)C 5 1.27-1.37 (m, 2H), 1.45 (s, 9H), .02 (m, 2H), 2.99-3.06 (m, 2H), 3.68-3.76 (m, 1H), 3.78 (s, 3H), .52 (m, 3H), 4.90 (s, 2H), 7.63-7.66 (m, 1H), 7.67-7.71 (m, 2H). [M+H] Calc’d for C24H27C1FN505, 520; Found, Preparation 15D: [2-(4-tert—butoxycarbonylamino-piperidin-l-yl)(4-cyano?uoro- phenyl)(4-methoxy-phenyl)oxo-6H-pyrimidin- l -yl]-acetic acid methyl ester ] A solution of [2-(4-tert-butoxycarbonylamino-piperidin-l-yl)chloro(4- cyano?uoro-phenyl)oxo-6H-pyrimidin-l-yl]-acetic acid methyl ester (2.2 g, 4.2 mmol), oxyboronic acid (1.9 g, 12.7 mmol), Pd-l 18 (274 mg, 0.42 mmol) and K2C03 (1.2 g, 8.4 mmol) in DMF (50 mL) was stirred at 145 0C for 6 h under nitrogen atmosphere. The reaction mixture was diluted with water and extracted with EA (3x).
The combined organics were washed with water, washed brine, dried (Na2S04) and concentrated. The residue was puri?ed by preparative HPLC to give 600 mg ofthe title product (24%). [M+H] Calc’d for C31H34FN506, 592; Found, 592.
Preparation 15E: 4-[2-(4-amino-piperidin-l-yl)-l-cyclopropylmethyloxo-l,6- dihydro-pyrimidinyl]?uoro-benzonitrile To a solution of [2-(4-tert-butoxycarbonylamino-piperidin-l-yl)(4-cyano ?uoro-phenyl)(4-methoxy-phenyl)oxo-6H-pyrimidin- l -yl]-acetic acid methyl ester (600 mg, 1.02 mrnol) in MeOH (10 mL) was added a 2N NaOH on (5 mL). After completion ofthe reaction, the solution was ed with lN HCl and extracted with EA (3x). The combined organics were washed with brine, dried (Na2S04) and concentrated. The residue was puri?ed by preparative HPLC to give 240 mg ofthe title product as a yellow solid (41%). [M+H] Calc’d for C30H32FN506, 578; Found, 578.
Example 15: [2-(4-amino-piperidin- l -yl)(4-cyano?uoro-phenyl)(4-methoxy- phenyl)oxo-6H-pyrimidin- l -yl] -acetic acid To a solution of [2-(4-tert-butoxycarbonylamino-piperidin-l-yl)(4-cyano ?uoro-phenyl)(4-methoxy-phenyl)oxo-6H-pyrimidin-l-yl]-acetic acid (100 mg, 0.15 mmol) in EA (10 mL) was added a 5N HCl solution in EA (5 mL). The reaction mixture was stirred at RT for 2 h, and the solvent was concentrated in vacuo. The residue was d by preparative HPLC to give 25 mg ofthe title product as HCl salt (32%). 1H NMR (400 MHz, : 5 1.53-1.56 (m, 2H), 2.00-2.03 (m, 2H), 3.00-3.07 (m, 2H), 3.35-3.39 (m, 1H), 3.67 (s, 3H), 4.70 (s, 2H),4.76-4.77 (m, 2H), 6.74 (d, J: 8.4 Hz, 2H), 6.96 (d, J: 8.8 Hz, 2 H), 7.17 (d, J: 8.4 Hz, 1H), 7.26 (d, J: 10.0 Hz, 1H), 7.50 (dd, J: 7.2, 8.0 Hz, 1H). [M+H] Calc’d for C25H24FN504, 478; Found, 478.
Preparation 16A: {l-[l-carbamoylmethyl(4-cyano?uoro-phenyl)(4-methoxy- phenyl)oxo- l ,6-dihydro-pyrimidinyl]-piperidinyl} mic acid tert—butyl ester To a solution of [2-(4-tert-butoxycarbonylamino-piperidinyl)(4-cyano ?uoro-phenyl)(4-methoxy-phenyl)oxo-6H-pyrimidinyl]-acetic acid (120 mg, 0.2 mmol) in DMF (5 mL) was added NH4Cl (17 mg HATU (95 mg,0.25 mmol) , 0.3mmol), and DIEA (25 mg, 0.4 mmol). After completion ofthe reaction, the solution was diluted with H20 and extracted with DCM for (3x). The combined organics were dried 4) and concentrated. The residue was puri?ed by ative HPLC to give 50 mg ofthe title product as a yellow solid (43%). [M+H] Calc’d for C30H33FN605, 577; Found, 577. e 16: 2-[2-(4-amino-piperidinyl)(4-cyano?uoro-phenyl)(4- methoxy-phenyl)oxo-6H-pyrimidinyl]-acetamide The title compound was prepared as the hydrochloride salt in 96 % yield according to the procedure for the preparation of Example 15. 1H NMR (400 MHz, CD30D)C 8 1.49-1.53 (m, 2H), 1.98-2.01 (m, 2H), 2.97-3.04 (m, 2H), .36 (m, 1H), 3.68 (s, 3H), 4.69 (s, 2H), 4.75-4.78 (m, 2H), 6.75 (d, J: 8.4 Hz, 2H), 6.99 (d, J: 8.8 Hz, 2 H), 7.16 (dd, J: 1.2, 8.0 Hz, 1H), 7.25 (dd, J: 0.8, 10.4 Hz, 1H), 7.49 (dd, J: 7.2, 8.0 Hz, 1H). [M+H] Calc’d for C25H25FN603, 477; Found, 477.
Preparation 17A: 2,6-dichloro(3-methoxy-propyl)-3H-pyrimidinone CI NTCI \/\/O\ To a solution of 2,6-dichloro-3H-pyrimidinone (600 mg, 3.65 mmol) in DMF (10 mL) was added K2C03 (1.0 g, 7.3 mmol) and the mixture was stirred at RT for min. 1-Bromomethoxy-propane (101 mg, 7.3 mrnol) was then added dropwise at 0 0C, and the mixture was d at RT overnight. DMF was concentrated in vacuo, and the residue was puri?ed by silica chromatography to give 400 mg of the title compound (47%). [M+H] Calc’d for; Calc’d for CngoClzNzOz, 237; Found, 237.
Preparation 17B: chloro(3-methoxy-propyl)oxo-1,6-dihydro-pyrimidin yl]-piperidinyl} -carbamic acid tert-butyl ester A on of 2,6-dichloro(3 -methoxy-propyl)-3H-pyrimidinone (400 mg, 1.68 mmol), dinyl-carbamic acid tert—butyl ester (405 mg, 2 mmol) and DIEA (260 mg, 2.0 mmol) in DMF (20 mL) was stirred at 85 0C for 2 h. The solvent was trated, and the residue was puri?ed by silica chromatography to give 500 mg of the title compound (75%). [M+H] Calc’d for C18H29C1N4O4, 400; Found, 400.
Preparation 17C: {1-[4-(4-cyano?uoro-phenyl)(3-methoxy-propyl)oxo-1,6- dihydro-pyrimidinyl]-piperidinyl} -carbamic acid tert—butyl ester \ H 0/ Bee N N | ‘Y N\/\/O\ A mixture of {1-[4-chloro(3 -hydroxy-propyl)oxo-1,6-dihydro-pyrimidin— 2-yl]-piperidinyl} -carbamic acid tert-butyl ester (200 mg, 0.5 mmol), 4-cyano ?uorophenyl boric acid (107 mg, 0.65 mmol), Pd(PPh3)4 (12 mg, 0.01 mmol) and 0.4M N32C03 solution (4 mL) in ACN was stirred at 85 0C overnight. The reaction muixture was diluted with water and extracted with EA (3x). The reaction mixture was stirred at RT for 2 h and the solvent was concentrated in vacuo. The e was puri?ed by silica chromatography to give 240 mg ofthe title product (99%). [M+H] Calc’d for C25H32FN504, 485; Found, 485.
Example 17: 4-[2-(4-amino-piperidinyl)(3-hydroxy-propyl)oxo-1,6-dihydro- pyrimidinyl]?uoro-benzonitrile ] To a solution of {1-[4-(4-cyano?uoro-phenyl)(3-methoxy-propyl)oxo- 1,6-dihydro-pyrimidin—2-yl]-piperidinyl} -carbamic acid tert—butyl ester (200 mg, 0.41 mmol) in DCM was added 1M BBr3 (4 mL) at -78 0C. The mixture was d at RT for 2 h and quenched at 0 0C with MeOH. The solution was washed with aqueous saturated NaHC03. The c layer was dried and concentrated. The residue was puri?ed by preparative HPLC to give 35 mg of the title product as the hydrochloride salt (23%). 1H NMR (400 MHz, CDgOD): 1.65-1.69 (m, 2H), 1.97-2.19 (m, 4H), 3.13-3.22 (m, 2H), 3.48-3.55 (m, 1H), 3.73 (t, J: 8.0 Hz, 2H), 4.55 (t, J: 8.0 Hz, 2H), 4.94-4.95 (m, 2H), 6.71 (s, 1H), 7.88-8.05 (m, 3H). [M+H] Calc’d for FN502, 371; Found, 371. ation 18A: {1-[5-benzo?aranyl(4-cyano?uoro-phenyl)methyloxo- 1,6dihydro-pyrimidinyl]-piperidinyl} -carbamic acid tert—butyl ester A mixture of {1-[5-chloro(4-cyano?uoro-phenyl)methyloxo-1,6- dihydro-pyrimidinyl]-piperidinyl} -carbamic acid tert—butyl ester (200 mg, 0.45 mmol), benzo?lranboronic acid (120 mg, 0.68 mmol), Pd(PPh3)4 (26 mg, 0.05 mmol) and 2M Na2C03 (0.9 mL) in 1,4-dioxane (200 mL) was re?uxed overnight under N2 atmosphere. The reaction mixture was diluted with water and extracted with EA (3x).
The combined organics were washed with brine, dried (Na2S04) and concentrated. The residue was puri?ed by silica chromatography to give 100 mg ofthe title product (42%).
[M+H] Calc’d for C30H30FN504, 543; Found, 543.
Example 18: 4-[2-(4-amino-piperidinyl)benzo?1ranyl— 1 -methyloxo- 1 ,6- dihydro-pyrimidinyl]?uoro-benzonitrile To a on of Preparation 18A (60 mg, 0.11 mmol) in EA (20 mL) was added a 4M HCl solution in EA (10 mL). The mixture was stirred at RT for 2h. The solvent was concentrated in vacuo to give 43 mg of the title t as the hydrochloride salt (53%).1H NMR (400 MHz, CD30D)C 1.85-1.92 (m, 2H), 2.13-2.18 (m, 2H), 3.10 (t, J: 4.0 Hz, 2H), 3.31-3.33 (m, 1H), 3.61 (s, 3H), 3.87 (d, J: 13.2 Hz, 2H), 6.65-7.21 (m, 3H), .76 (m, 4H), 7.76 (s, 1H). [M+H] Calc’d for C25H22FN502, 443; Found, 443.
Preperation 19A: {l-[5-cyano(4-cyano?uoro-phenyl)- l -methyloxo- l ,6- dihydro-pyrimidinyl] -piperidinyl} -carbamic acid tert—butyl ester A mixture of {l-[5-chloro(4-cyano?uoro-phenyl)-l-methyloxo-1,6- dihydro-pyrimidinyl]-piperidinyl} -carbamic acid tert—butyl ester (460 mg, 1 mmol), Zn(CN)2 (175 mg 1.5 mmol) and Pd(PPh3)4 (l 16 mg, 0.0.1 mmol) in DMF (5 mL) was stirred for 4 h at 150 0C under N2 atmosphere. The reaction mixture was cooled to RT and d. The ?ltrate was concentrated in vacuo, and the residue was puri?ed by preparative HPLC to give 150 mg ofthe title product as a yellow solid (33%). [M+H] Calc’d for FN603, 453; Found, 453.
Example 19: 2-(4-amino-piperidin- l -yl)(4-cyano ro-phenyl)- l -methyloxo- l ,6-dihydro-pyrimidine-5 -carbonitrile To a solution of {l-[5-cyano(4-cyano?uoro-phenyl)-l-methyloxo-l,6- dihydro-pyrimidinyl]-piperidinyl} -carbamic acid tert—butyl ester (150 mg, 0.33 mmol) in EA (5 mL) was added a 5N HCl solution in EA (5 mL). The reaction mixture was stirred at RT for 2 h, and the solvent was concentrated in vacuo to give 120 mg of the title product as HCl salt (94%). 1H NMR (400 MHz, CDgOD): 8 1.67-1.72 (m, 2H), 2.02-2.06 (m, 2H), 3.13-3.16 (m, 2H), 3.34-3.38 (m, 1H), 3.42 (s, 3H), .02 (m, 2H), 7.82-7.90 (m, 3H). [M+H] Calc’d for C18H17FN6O, 353; Found, 353.
Example 20: 4-[2-(4-aminopiperidin- l -yl)-5 -chloro - l loxopyrimidinyl] ?uorobenzonitrile NC NH2 CI \ To a solution of {1-[5-chloro(4-cyano?uoro-phenyl)methyloxo-1,6- dihydro-pyrimidinyl]-piperidinyl} -carbamic acid tert—butyl ester (150 mg, 0.33 mmol) in EA (5 mL) was added a 5N HCl solution in EA (5 mL). The reaction mixture was stirred at RT for 2 h, and the solvent was concentrated in vacuo to give 120 mg of the title t as HCl salt (94%). 1H NMR (400 MHz, : 8 1.67-1.72 (m, 2H), 2.02-2.06 (m, 2H), 3.13-3.16 (m, 2H), 3.34-3.38 (m, 1H), 3.42 (s, 3H), 3.98-4.02 (m, 2H), 7.82-7.90 (m, 3H). [M+H] Calc’d for C18H17FN6O, 353; Found, 353. 1H NMR (400 MHz, METHANOL-d4): 8 ppm 1.73 - 1.91 (m, 2 H), 2.18 (d, J=12.13 Hz, 2 H), 3.06 (t, J=12.76 Hz, 2 H), 3.33 - 3.40 (m, 1 H), 3.57 (s, 3 H), 3.83 (d, J=13.14 Hz, 2 H), 7.75 - 7.93 (m, 3 H). ( 2, CDgOD): 8 1.89-1.93 (m, 2H), 2.18-2.21 (m, 2H), 2.73 (s, 3H), 2.74 (s, 3H), 3.11-3.17 (m, 2H), 3.33- 3.39 (m, 1H), 3.57 (s, 3H), 34-397 (m, 2H), 7.28 (d, J = 8.0 Hz, 1H), 7.55 (d, .1: Prepared by the ure of Example‘ 1 .0 Hz, 1H), 7.63-7.67 (m, 1H), 7.74 (d, J: 8.4 Hz, 1H), 8.12 (d, J: 8.4 Hz, 1H), 8.65 (s, 1H).
H NMR (400 MHz, CDClg): 8 1.74-1.80 (m, 4H), 1.93-1.97 (m, 2H), 3.11 (s, 2H), 3.26-3.35 (m, 6H), 3.47 (s, 3H), 3.75 (s, 3H), 6.68 (dd, J: 1.2, 8.4 Hz, 1H), 6.86-6.72 (m, 2H), 7.11 (dd, J: 1.2, 8.0 Hz, 1H), 7.30 (de= 1.2, Prepared by the procedure of Example 1 10.8 Hz, 1H), 7.46 (de = 6.8, 7.6 Hz, 1H .
H NMR (400 MHz, CD30D): .82 (m, 2H), 2.03-2.06 (m, 2H), 3.06-3.12(1n, 2H), 3.22— 3.34 (m, 1H), 3.49 (s, 3H), 3.1 (d, .1: 13.6 Hz, 2H), 7.07—7.09 (m, 1H), 7.36— 7.38 (m, 1H), 7.51—7.55 (m, 1H), 7.66 (d, .1: 8.0 Hz, 1H), 7.79—7.82 (m, 1H), 8.33 (s, 1H).
H NMR (400 MHz, CD30D): 1.96-2.01(rn, 2H), 2.20—2.22 (m, 2H), 3.23—3.32 (m, 2H), 3.46- 3.49 (m, 1H), 3.65 (s, 3H), 3.94—3.97 (m, 2H), 4.39 (s, 3H), 7.55—7.77 (m, 7H), 8.76 (s, 1H).
H NMR (300 MHz, CD30D): 5 1.72—1.93 (m, 3H), 1.97—2.23 (m, 1H), 3.16-3.30(1n, 2H), 3.50— 3.55 (m, 2H), 3.60 (s, 3H), 3.83—3.84 (m, 1H), 3.86 (s, 3H), 6.82 (d, .1: 8.1 Hz, 1H), .05 H), 7.25 (d, .1: 8.1 Hz, 1H), 7.44 (d, .1: 10.8 Hz, 1H), H NMR (400 MHz, CDgOD): 1.64-1.69(1n, 2H), 1.89-1.92 (m, 2H), 2.85-2.91 (111,2H),3.15- 3.20 (m, 1H), 3.34 (s, 3H), 3.62 (d, .1: 8.4 Hz, 2H), 3.71 (s, 3H), 6.99 (d, .1: 8.4 Hz, 1H),7.20-7.40(1n, Pm oarcd bv the ?rmedure OfEXEiE‘YII a 4H).
H NMR (400 MHz, CDgOD): 5 2.19—2.22 (m, 1H),2.49-2.51 (m, 1H), 3.63 (s, 3H), 3.75-3.81 (m, 2H), 3.87 (s, 3H), 3.87— 3.93 (m, 1H), 4.02—4.06 (m, 2H), 6.80 (d, .1: 8.4 Hz, 1H), 7.00 (t, .1: 10.8 Hz, 2H), 7.25 (d, .1: 9.6 Hz, 1H), 7.44 (d, .1: 10.8 Hz, 1H), 7.61 (t, .1: 7.4 H NMR (400 MHz, CDgOD): 5 1.69-1.99 (m, 3H),2.14-2.19(rn, 1H), .24 (m, 2H), 3.43— 3.46 (m, 1H), 3.56—3.60 28 (m, 4H), 3.77—3.80 (m, 1H), 3.82 (s, 3H), 6.77 (d, .1: 8.0 Hz, 1H), 6.94-7.00 (m, 2H), 7.21 (d, .1: 8.0 Hz, 1H), 7.39 (d, .1: 10.4 Hz, 1H), 8.56-7.60 (m, H NMR (400 MHz, CDgOD): 5 2.25—2.29 (m, 1H), 2.50—2.55 (m, 1H), 3.69 (s, 3H), 3.89—3.84 (m, 5H), 3.99—4.03 (m, 1H), 29 5.05—4.16 (m, 2H), 6.80 (d, .1: 8.4 Hz, 1H), .03 (m, 2H), 7.29 (dd, .1: 2.4, 8.0 Hz, 1H), 7.47 (d, .1: .4 Hz, 1H), 7.64 (dd, .1: 6.8, 8.0 Hz, 1H. 1H NMR (400 MHz, CDgOD): 5 1.73—2.02 (m, 3H), 2.19—2.23 (m, 1H), 3.13—3.26 (m, 2H), 3.49— 3.52 (m, 2H), 3.60 (s, 3H), 3.77-3.85 (m, 1H), 3.85 (s, 3H), 6.89 (d, .1: 11.6 Hz, 2H), .10 (d, .1: 11.6 Prepared? by the prmedm? 0f Eixampie 3 Hz, 2H), 7.24 (d, .1: 8.0 Hz, 1H), 7.41 (d, .1: 10.8 Hz, 1H), 7.57-7.61 (m, H NMR (400 MHz, CDgOD): 8 .99 (m, 3H), 2.07-2.10 (m, 1H), 3.09-3.24 (m, 2H), 3.43- 3.46 (m, 1H), 3.56-3.60 (m, 4H), 3.68 (s, 3H), 31 3.76-3.79 (m, 1H), 6.75 (d, J: 8.8 Hz, 2H), 6.96 (d, J Prepared by the procedure if Exampie 1 = 9.2 Hz, 2H), 7.13 (dd, J = 2.0, 8.0 Hz, 1H), 7.27 (dd, J: 0.8, 10.4 Hz, 1H), 7.47 (dd, J: 6.8, 8.0 Hz, H NMR (400 MHz, CDgOD): 8 1.41 (s, 3H), 1.82-1.85(rn,2H),1.91- 1.99 (m, 2H), 3.22-3.25 (m, 2H), 3.47 (s, 3H), 3.50- 32 3.57 (m, 2H), 3.75 (s, 3H), 6.69 (d, J: 8.4 Hz, 1H), 6.86-6.92 (m, 2H), 7.14 (d, J: 8.4 Hz, 1H), 7.32 (d, J = 10.8 Hz, 1H), 7.47-7.51 m, 1H .
H NMR (400 MHz, CD3OD): 8 1.95-1.99 (m, 2H), 2.19-2.22 (m, 2H), 3.20-3.26 (m, 2H), 3.45- 33 3.50 (m, 1H), 3.63 (s, 3H), 3.90 (d, J: 12.8 Hz, 2H), 4.06 (s, 3H), 7.21 (d, J: 8.4 Hz, 1H), 7.48-7.57 (m, 6H), 7.96 (s, 1H).
H NMR (400 MHz, CDgOD): 8 1.75-1.79 (m, 2H), 2.02-2.05 (m, 2H), .06 (m, 2H), 3.21- 3.31 (m, 1H), 3.48 (s, 3H), 3.72-3.75 (m, 2H), 4.77- 4.81 (m, 2H), 7.22 (s, 1H), 7.31 (d, J: 8.0 Hz, 1H), 7.39 (d, J: 2.0 Hz, 1H), Prepared by the ure if Exampie 1 7.60-7.64 m, 2H .
H NMR (400 MHz, CD30D): 1.85-1.99 (m, 2H), 2.18-2.20 (m, 2H), 3.19—3.24 (m, 2H), 3.46- 3.50 (m, 1H), 3.86 (s, 3H), 457 3.86-3.92 (m, 2H), 4.10 (s, 3H), 7.21—7.25 (m, 2H), 7.40—7.53 (m, 4H), 8.01 (s, H NMR (400 MHz, CDgOD): 8 1.79-1.82 (m, 2H), 2.04-2.07 (m, 2H), 3.09-3.15 (m, 2H), 3.32- 3.38 (m, 1H), 3.50 (s, 3H), 36 458 .79 (m, 2H), 4.74- 4.78 (m, 2H), 7.12 (s, 1H), 7.55 (d, J: 8.4 Hz, 2H), 7.62 (s, 1H), 7.67 (d, J: 8.0 Hz, 2H).
H NMR (400 MHz, CDgOD): 8 1.98-2.04 (m, 2H), 2.21-2.24 (m, 2H), 3.27-3.30 (m, 2H), 3.50- 3.52 (m, 1H), 3.65 (s, 3H), 37 458 3.98 (d, J: 12.8 Hz, 2H), 4.42 (s, 3H), 7.33 (d, J: 8.0 Hz, 1H), 7.49 (d, J: .0 Hz, 1H), 7.60-7.73 (m, 3H), 7.84 (s, 1H), 8.85 s, 1H .
H NMR (400 MHz, : 1.87-1.94 (m, 2H), 2.15 (d, J: 12.0 Hz, 2H), 3.13 (t, J: 8.4 Hz, 2H), 3.39-3.43 (m, 1H), 38 452 3.59 (s, 3H), 3.87 (d, J: 12.8 Hz, 2H), 3.97 (s, 3H), 6.79 (d, J: 8.8 Hz, 2H), 7.55 (d, J: 8.4 Hz, 2H), Prapa?‘ed by the pmcedure Hfiixampie 1 7.70 d, J: 8.4 Hz, 2H .
H NMR (400 MHz, CD30D):81.91-1.94(m, 2H), 2.16-2.19 (m, 2H), 3.15-3.21 (m, 2H), 3.50- 3.52 (m, 1H), 3.61 (s, 3H), 39 448 3.90 (d, J: 12.4 Hz, 2H), 7.22 (d, J: 8.0 Hz, 1H), 7.30 (d, J: 8.0 Hz, 2H), 7.43 (d, J: 10.8 Hz, 1H), 7.59 (t, J: 7.2 Hz, 1H), 7.86 (d, J: 8.0 Hz, 2H).
H NMR (400 MHz, CD30D):81.88-1.91(m, 2H), 2.12-2.13 (m, 2H), 2.94 (s, 3H), 3.13-3.15 (m, 2H), 3.30-3.34 (m, 1H), 40 461 3.61 (s, 3H), 3.89 (d, J: 14.4 Hz, 2H), 7.00 (d, J: 8.0 Hz, 1H), 7.11 (d, J: 12.0 Hz, 1H), 7.53 (d, J: 12.0 Hz, 2H), .64 m, 3H .
H NMR (400 MHz, CDgOD): 8 1.87-1.91 (m, 2H), 2.14-2.16 (m, 2H), 3.15 (t, J: 12.0 Hz, 2H), 41 447 3.30-3.40 (m, 1H), 3.61 (s, 3H), 3.89 (d, J: 14.0 Hz, 2H), 7.01(d, J: 8.0 Hz, 1H), 7.13 (d, J: 12.0 Hz, 1H), 7.52-7.77 (m, 5H).
H NMR (400 MHz, CDgOD): 8 79 (m, 2H), 2.03—2.05 (m, 2H), 3.00-3.06 (m, 2H), 3.21- 3.31 (m, 1H), 3.49 (s, 3H), 3.75-3.78 (m, 2H), 4.32 (s, 42 459 2H), 7.06 (dd, J: 1.2, 8.0 Hz, 1H), 7.12 (dd, J=1.2, 8.0 Hz, 1H), 7.31-7.36 (m, Prepared by the pi‘dcedure of Exampk: 1 2H), 7.42 (dd, J: 6.4, 7.6 Hz, 1H), 7.58 (d, .1: 7.6 Hz, 1H).
H NMR (400 MHz, CDgOD): 5 .80 (m, 2H), 2.02—2.05 (m, 2H), 3.01—3.05 (m, 2H), 3.35— 3.36 (m, 1H), 3.49 (s, 3H), 43 448 3.74—3.98 (m, 2H), 7.07 (dd, .1: 1.6, 8.4 Hz, 1H), 7.27—7.32 (m, 3H), 7.44 (dd, .1: 6.8, 8.0 Hz, 1H), 7.73 (d, .1: 1.2 Hz, 1H), 7.84 (d, .1: 7.2 Hz, 1H).
H NMR (400 MHz, CDgOD): 5 1.87—1.91 (m, 1H), 2.25-2.28 (m, 1H), 2.87—2.92 (m, 1H), 3.11— 3.17 (m, 1H), 3.30—3.32 44 434 (m, 1H), 3.41—3.56 (m, 5H), 3.69—3.71 (m, 2H), 3.84 (s, 3H), 6.75 (d, .1: 8.4 Hz, 1H), .96 (m, Prepared by the proced 1m: 611’ 588.1111in 3 2H), 7.53 (d, .1: 8.0 Hz, 2H), 7.66 (d, .1: 8.4 Hz, H NMR (400 MHz, CDgOD): 5 1.74—1.80 (m, 1H),2.14-2.19(m, 1H), 2.77—2.81(m, 1H), 3.01— 3.06 (m, 1H), 3.31—3.34 45 434 (m, 1H), 3.36-3.45 (m, 5H), 3.59-3.60 (m, 2H) 3.71 (s, 3H), 6.63 (d, .1: 8.4 Hz, 1H), 6.80—6.84 (m, 2H), 7.44 (d, .1: 8.0 Hz, 2H), 7.60 (d, .1: 8.4 Hz, 1H NMR (400 MHz, : 5 2.15-2.18 (m, 2H), 3.31—3.34 (m, 2H), 3.46—3.51 (m, 5H), 3.56- 3.59 (m, 2H), 3.74 (s, 3H), 3.78—3.81 (m, 2H), 6.68 46 452 (dd, .1: 1.2, 8.4 Hz, 1H), 6.85—6.89 (m, 2H), 7.12 (dd, .1: 1.2, 7.6 Hz, 1H), Prapa?‘ed by the pmcedure ufElxampie I 7.28 (dd, .1: 1.6, 10.8 Hz, 1H), 7.47 (dd, .1: 6.8, 8.0 Hz, 1H .
H NMR (400 MHz, DMSO-d6): 8 3.27-3.34 (m, 4H), 3.45 (s, 3H), 3.51- 3.53 (m, 4H), 3.81 (s, 3H), 6.78 (d, J: 8.4 Hz, 1H), 47 438 7.02-7.08 (m, 2H), 7.18 (dd, J: 1.6, 8.4 Hz, 1H), 7.45 (dd, J: 1.6, 10.8 Hz, 1H), 7.80 (dd, J: 7.2, 8.0 Hz, 1H), 9.41 (br, 1H).
H NMR (400 MHz, CDgOD): 8 184-194 (m, 2H), 2.20-2.23 (m, 2H), 3.00-3.07 (m, 2H), 3.38- 3.42 (m, 5H), 3.72 (s, 3H), 48 434 4.22-4.27 (m, 1H), 6.61 (d, J: 8.8 Hz, 1H), 6.79-6.83 (m, 2H), 7.39 (d, J: 8.0 Hz, 2H), 7.51 (d, J: 8.0 Hz, 2H).
H NMR (400 MHz, CDgOD): 8 1.82-1.87 (m, 2H), 2.04-2.07 (m, 2H), 3.06-3.12 (m, 2H), 3.25 (s, 49 449 6H), 3.28-3.39 (m, 1H), 3.49 (s, 3H), 3.81-3.84 (m, 2H), 7.37 (d, J: 8.0 Hz, 1H), 7.56 (d, J: 9.6 Hz, 1H), 7.42 (t, J: 6.8 Hz, 1H), 8.31 (s, 2H).
H NMR (400 MHz, CDgOD): 8 1.93-1.97 (m, 2H), 2.17-2.20 (m, 2H), 3.03 (s, 3H), 3.20-3.26 (m, 2H), .53 (m, 1H), 50 462 3.62 (s, 3H), 3.98-4.02 (m, 2H), 7.32 (d, J: 8.0 Hz, 1H), 7.60 (d, J: 10.0 Hz, 1H), 7.67 (t, J: 6.4 Hz, Prepared by the pzmedure ofExmane 1 1H), 8.32 (s, 2H), 8.83 (s, H NMR (400 MHz, CDgOD): 8 1.89-1.91 (m, 1H), 2.26-2.28 (m, 1H), 2.91-2.93 (m, 1H), 3.12- 3.15 (m, 1H), 3.30-3.32 (m, 1H), 3.42-3.55 (m, 5H), .72 (m, 2H) 3.84 (s, 3H), 6.84 (d, J: Prepared by the procedure 61" Exampie 1 8.4 Hz, 2H), 7.03 (d, J 8.4 Hz, 2H), 7.29 (d, J 8.0 Hz, 1H), 7.39 (d, J .0 Hz, 1H), 7.64 (t, J: 7.2 Hz, 1H).
H NMR (400 MHz, CDgOD): 8 1.86-1.91 (m, 1H), 2.22-2.28 (m, 1H), 2.97-2.91 (m, 1H), 3.10- 3.13 (m, 1H), 3.29-3.32 (m, 1H), 3.40-3.51 (m, 5H), 3.67-3.69 (m, 2H) 3.82 (s, 3H), 6.84 (d, J: Prepared by the pmcedure pie 1 8.0 Hz, 2H), 7.02 (d, J: 8.4 Hz, 2H), 7.27 (d, J: 8.0 Hz, 1H), 7.35 (d, J: .4 Hz, 1H), 7.64 (t, J: 7.2 Hz, 1H .
H NMR (400 MHz, CDgOD): 8 2.03-2.06 (m, 2H), 2.32-2.35 (m, 2H), 3.14-3.21(rn,2H),3.51- 3.56 (m, 5H), 3.78 (s, 3H), 4.37-4.39 (m, 1H), 6.84 (d, J: 7.2 Hz, 2H), 7.02 (d, J = 8.0 Hz, 2H), 7.27 (d, J: eci by the pra‘medur? 0f Eixampie 1 8.0 Hz, 1H), 7.38 (d, J: .4 Hz, 1H), 7.62 (t, J: 7.2 Hz, 1H).
H NMR (400 MHz, CDgOD): 8 1.66-1.71 (m, 1H), 2.11-2.16 (m, 1H), 2.77-2.81 (m, 1H), 2.93- 2.97 (m, 4H), 3.16-3.20 (m, 1H), 3.30-3.38 (m, 54 448 2H), 3.43-3.50 (m, 5H), 3.69 (s, 3H), 6.75 (d, J: Prepared by the procedure 61" e 1 8.4 Hz, 2H), 6.95 (d, J: 8.4 Hz, 2H), 7.16 (d, J: 8.4 Hz, 1H), 7.28 (d, J: .8 Hz, 1H), 7.50 (dd, J: 6.8, 8.0 Hz, 1H).
H NMR (400 MHz, CDgOD): 8 2.03-2.13 (m, 4H), 2.84 (s, 3H), 3.01- 3.05 (m, 2H), 3.39-3.43 (m, 2H), 3.48 (s, 3H), 3.67 55 448 (s, 3H), 3.87-3.92 (m, 1H), 6.74 (d, J: 8.8 Hz, 2H), 6.95 (d, J: 8.8 Hz, 2H), 7.13 (dd, J: 1.2, 8.0 Hz, 1H), 7.21 (dd,.]= 1.6, 10.4 Hz, 1H), 7.45 (dd, J: 6.8, 7.6 Hz, 1H).
H NMR (400 MHz, CDgOD): 8 1.77-1.80 (m, 1H), 2.22-2.26 (m, 1H), 2.90-2.92 (m, 1H), 3.03- 3.07 (m, 4H), 3.27-3.30 (m, 1H), 3.39-3.41 (m, 56 448 2H), 3.44-3.46 (m, 5H), 3.77 (s, 3H), 6.86 (d, J: Prepared by the re ofExmane 1 8.4 Hz, 2H), 7.06 (d, J: 8.0 Hz, 2H), 7.29 (d, J: 8.4 Hz, 1H), 7.36 (d, J: 8.4 Hz, 1H), 7.58 (dd, J: 6.8, 7.6 Hz, 1H).
H NMR (400 MHz, CDgOD): 8 1.77-1.85 (m, 2H), 2.03-2.06 (m, 2H), 3.03-3.09 (m, 2H), 3.18 (s, 6H), 3.31-3.38 (m, 1H), 57 448 3.48 (s, 3H), 3.78-3.81 (m, 2H), 7.03 (d, J: 9.2 Hz, 1H), 7.28 (d, J: 7.6 Hz, 1H), 7.46 (d, J: 10.0 Hz, Prepared by the pi‘dcedure of Exampk: 1 1H), 7.59-7.63 (m, 2H), s, 1H .
H NMR (400 MHz, CD30D):51.83-1.88(rn, 2H), 2.21—2.24 (m, 2H), 2.77 (s, 3H), 3.06-3.14 (m, 2H), 3.31—3.32 (m, 1H), 3.58 (s, 3H), 3.87-3.91 (m, 58 449 5H), 6.83 (d, .1: 11.2 Hz, 1H), 7.22 (dd, J: 2.0, 10.8 Hz, 1H), 7.42 (dd, .1: 2.0, 14.4 HZ, 1H), 7.59-7.65 (m, 2H), 7.84 (d, J: 3.2 Hz, 1H).
H NMR (400 MHz, CDgOD): 8 1.88-1.89 (m, 2H), 2.14—2.19 (m, 2H), 3.13-3.19 (m, 2H), 3.28 (s, 59 447 6H), 3.41-3.46 (m, 1H), 3.60 (s, 3H), 3.87-3.91 (m, 2H), 7.24 (d, J: 10.8 Hz, 1H), .44 (m, 3H), 7.59-7.67 (m, 3H).
H NMR (400 MHz, CD30D):81.91-1.97(m, 2H), 2.16-2.20 (m, 6H), 3.14—3.20 (m, 2H), 3.47— 3.49 (m, 1H), 3.60-3.63 474 (m, 7H), 3.89-3.92 (m, 2H), 7.31 (d, .1: 9.6 Hz, 1H), 7.01 (d, .1: 9.6 Hz, 1H), 7.41 (d, .1: 8.0 Hz, 1H), 7.58 (d, .1: 10.0 Hz, 1H), 7.68-7.75 (111,2H), 7.79 (s, 1H).
H NMR (400 MHz, CD30D): 5 2.27—2.30 (m, 2H), 344—347 (m, 2H), 3.60-3.64 (m, 5H), 3.70— 3.73 (m, 2H), 3.91—3.94 61 435 (m, 5H), 6.83 (d, .1: 8.4 Hz, 1H), 7.24 (dd, .1: 1.6, 8.0 Hz, 1H), 7.43 (dd, .1: Prepared by the procedure 61‘," Exampie 1 1.2, 10.4 Hz, 1H), 7.59— 7.66 (m, 2H), 7.84 (d, .1: 2.4 Hz, 1H .
H N MR (400 MHz, CDgOD): 8 .31 (m, 2H), 344—347 (m, 2H), 3.60—3.64 (m, 5H), 3.70— 3.73 (m, 2H), 3.91—3.94 62 417 (m, 5H), 6.81 (d, .1: 8.4 Hz, 1H), 7.53 (d, .1: 8.4 Hz, 2H), 7.58 (dd, .1: 2.4, 8.8 Hz, 1H), 7.64 (d, .1: 8.8 Hz, 2H), 7.84 (d, .1: 1.2 Hz, 1H).
H NMR (400 MHz, CDgOD): 8 2.31—2.33 (m, 2H), 3.27 (s, 6H) 344—347 (m, 2H), 3.60—3.67 (m, 5H), 3.73—3.76 (m, 2H), 63 448 3.96—3.99 (m, 2H), 6.81 (d, .1: 8.4 Hz, 1H), 7.53 (d, .1 = 8.4 Hz, 1H), 7.58 (dd, .1 = 2.4, 8.8 Hz, 1H), 7.64 (d, .1: 8.8 Hz, 2H), 7.84 (d, .1 = 1.2 Hz, 1H H NMR (400 MHz, CD30D): 8 3.39 (s, 3H), 3.67 (s, 3H), 4.09—4.10 (m, 1H), 4.17—4.21 (m, 2H), 4.55—4.59 (m, 2H), 6.74 (d, 64 406 .1: 8.8 Hz, 2H), 6.96 (d, .1 = 8.8 Hz, 2H), 7.10 (dd, .1 = 1.6, 8.4 Hz, 1H), 7.23 (dd, .1: 1.6, 10.8 Hz, 1H), 7.45 (dd, .1: 6.8, 8.0 Hz, 1H NMR (400 MHz, CDgOD): 8 .98 (m, 2H), 2.23—2.27 (m, 2H), 2.78 (s, 3H), 3.38—3.40 (m, 1H), 3.62 (s, 3H), 3.90— 65 472 3.95 (m, 2H), 4.41 (s, 3H), 7.26 (d, .1: 10.8 Hz, 1H), 7.40 (d, .1: 13.6 Hz, 1H), Prepared by the pmcedure ofExampie 1 7.49—7.57 (m, 2H), 7.65 (dd, .1: 6.8, 11.6 Hz, 1H), 7.73 (s, 1H), 8.66 (s, 1H). l H NMR (400 MHz, CDgOD): 8 2.19-2.20 (m, 2H), 3.33-3.36 (m, 2H), 3.50-3.53 (m, 5H), 3.60- 3.63 (m, 2H), 3.83-3.85 458 (m, 2H), 4.42 (s, 3H), 7.13 (d, J: 7.6 Hz, 1H), 7.28 (d, J: 10.4 Hz, 1H), 7.34 (d, J: 9.2 Hz, 1H), 7.40 (t, J: 7.2 Hz, 1H), 7.52 (d, J = 8.8 Hz, 1H), 7.58 (s, 1H), 8.48 (s, 1H).
H NMR (400 MHz, CDgOD): 8 2.33-2.35 (m, 2H), 3.27 (s, 6H), 3.46- 3.49 (m, 2H), 3.62-3.66 430 (m, 5H), 3.75-3.78 (m, 2H), 3.98-4.02 (m, 2H), 6.73 (d, J: 9.2 Hz, 1H), 7.67-7.72 (m, 5H), 7.80 (s, l H NMR (400 MHz, CD30D): 5 .97 (m, 2H), 2.15-2.18 (m, 2H), .21 (m, 2H), 3.43— 3.49 (m, 1H), 3.61 (s, 3H), 490 3.69-3.71 (m, 4H), 3.86- 3.94 (m, 6H), 7.31 (d, .1: 9.6 Hz, 1H), 7.41 (d, .1: 8.0 Hz, 1H), 7.59 (d, .1: .0 Hz, 1H), 7.72-7.80 (m, 2H), 7.91 (s, 1H).
H NMR (400 MHz, CD30D): 5 2.99-3.06 (m, 1H), 3.30—3.32 (m, 2H), 3.49 (s, 3H), 3.78 (s, 3H), 4.10—4.15 (m, 2H), 4.46- 420 4.52 (m, 2H), 6.83 (d, .1: 8.4 Hz, 2H), 7.01 (d, .1: 8.8 Hz, 2H), 7.19 (dd, .1: Prepared by the pmcedure of EXEi?‘Ep1C 1 2.0, 10.8 Hz, 1H), 7.33 (dd, J: 2.0, 14.4 Hz, 1H), 7.45 (dd,.]= 88,108 Hz, 1H).
H NMR (400 MHz, CDgOD): 8 2.78 (s, 3H), 3.07-3.10(rn, 1H), 3.37— 3.39 (m, 2H), 3.48 (s, 3H), 3.78 (s, 3H), 4.12—4.15 (m, 70 2H), 4.47—4.52 (m, 2H), 6.84 (d, .1: 8.8 Hz, 2H), 7.01 (d, .1: 8.8 Hz, 2H), 7.20 (d, .1: 8.0 Hz, 1H), 7.33 (d, .1: 10.4 Hz, 1H), 7.45 (t, .1: 7.2 Hz, 1H).
H NMR (400 MHz, CDgOD): 5 1.67-1.73 , 2.02-2.06 (m, 2H), 2.33 (s, 6H), 2.41—2.45 (m, 1H), 2.95—3.03 (m, 2H), 3.59 (s, 3H),3.79—3.84 (m, 2H), 4.18 (s, 3H), 7.10 (dd, .1: 2.0, 12.0 Hz, 1H), 7.23 (dd, .1: 1.6, 10.8 Hz, 1H), 7.37 (dd, .1: 2.0, 14.4 Hz, 1H), 7.46-7.56 (m, 3H), 8.11 (s, 1H).
H NMR (400 MHz, DMSO-d6): 5 1.50-1.63 (m, 2H), 1.85-1.89 (m, 2H), 2.18 (s, 6H), 2.21— 2.27 (m, 1H), 2.85-2.92 72 (m, 2H), 3.45 (s, 3H), 3.67- 3.71 (m, 2H), 4.00 (s, 3H), 7.09-7.17(rn, 2H), 7.39 Prepared? by the prmedur? 0f ie 3 (dd, .1: 1.6, 14.4 Hz, 1H), 7.51—7.54 (m, 2H), 7.69 (t, .1: 9.2 Hz, 1H), 7.97 (s, 1H NMR (300 MHz, :81.85-1.91(m, 2H), 2.11-2.16 (m, 2H), 3.06-3.14 (m, 2H), 3.36- 3.40 (m, 1H), 3.57 (s, 3H), 3.81-3.85 (m, 2H), 6.85 (d, .1: 8.4 Hz, 2H), 7.22 (d, .1 = 3.0 Hz, 1H), 7.24 (s, 1H), 7.33—7.48 (m, 4H).
Prepared by the procedure of Exampie 18 H NMR (400 MHz, CD30D): 5 1.53-1.56 (m, 2H), 1.88-1.91 (m, 2H), 2.87-2.95 (m, 3H), 3.49 (s,3H), 3.62 (d, .1: 13.6 Hz, 2H), 3.69 (s, 3H), 6.26 (s, 1H), 6.81 (d, .1: 4.0 Hz, 1H), .35 (m, 6H).
H NMR (400 MHz, CDgOD): 8 1.88-1.94 (m, 2H), 2.13-2.16 (m, 2H), 3.05-3.16 (m, 2H), 3.33- 3.42 (m, 1H), 3.60 (s, 3H), 3.84-3.86 (m, 2H), 6.74- 6.77 (m, 1H), 7.20-7.50 (m, 7H).
H NMR (400 MHz, CD30D):81.51-1.54(rn, 2H), 1.88-1.91 (m, 2H), 2.84-2.97 (m, 3H), 3.49 (s, 3H), 3.62-3.65 (m, 5H), 6.31 (d, .1: 2.8 Hz, 1H), 6.61 (d, .1: 8.0 Hz, 1H), 7.06 (d, .1: 3.2 Hz, 1H), 7.13-7.18 (m, 3H), 7.27 (d, J: 10.8 HZ, 1H), 7.33-7.37 (m, 2H).
H NMR (400 MHz, CD3OD): 5 1.65-1.68 (1’11, 2H), 2.01—2.04 (m, 2H), 2.98-3.12 (m, 3H), 3.63 (s, 3H), 3.78-3.82 (m, 2H), 6.94 (d, .1: 8.4 Hz, 1H), 7.24 (d, .1: 8.0 Hz, 1H), 7.43-7.45 (m, 2H), 7.50 (t, J: 7.2 Hz, 1H), 7.72 (d, .1 Prepared by the pmmedure ofExampEe 1 = 8-4 HZ, 1H), 8-06 (S, 1H)- H NMR (300 MHz, CDgOD): 8 1.84-1.91 (m, 1H), 2.01-2.06 (m, 1H), .08 (m, 2H), 3.16- 3.21 (m, 1H), 3.58 (s, 3H), 3.75-3.82 (m, 4H), 3.93- 78 4.01 (m, 1H), .82 (m, 1H), 6.86 (d, J: 9.0 Hz, 2H), 7.06 (d, J: 8.7 Prapa’d‘ed by the pmcedure dfliixampie 1 Hz, 2H), 7.24 (dd, J: 0.9, 8.1 Hz, 1H), 7.34 (dd, J: 1.5, 10.8 Hz, 1H), 7.54- 7.58 (m, 1H).
H NMR (400 MHz, CDgOD): 8 1.87-1.91 (m, 1H), 2.03-2.07 (m, 1H), 3.02-3.08 (m, 2H), 3.19- 3.29 (m, 1H), 3.59 (s, 3H), 3.77-3.83 (m, 4H), 3.95- 79 4.01 (m, 1H), 4.73-4.85 (m, 1H), 6.87 (d, J: 8.8 Hz, 2H), 7.07 (d, J: 8.8 Prepared by the pzmedure ofExmane 1 Hz, 2H), 7.26 (dd, J: 1.2, 8.4 Hz, 1H), 7.36 (dd, J: 1.2, 10.8 Hz, 1H), 7.56 (t, J = 6.8 Hz, 1H H NMR (400 MHz, CD30D):81.71-1.77(m, 2H), 2.05-2.08 (m, 2H), 2.38 (s, 6H), 2.45-2.48 (m, 1H), 2.98-3.05 (m, 2H), 3.61 (s, 3H), 3.83-3.86 (m, 2H), 4.20 (s, 3H), 6.93 (dd, J: 1.2, 8.8 Hz, 1H), 7.27 (dd, J: 1.2, 7.6 Hz, 1H), .41 (m, 2H), 7.49- 7.53 (m, 1H), 7.66 (d, J: 8.8 Hz, 1H), 8.18 (s, 1H).
H NMR (400 MHz, CDgOD): 8 2.03-2.06 (m, 2H), 2.25-2.27 (m, 2H), 2.98 (s, 6H), 3.14-3.20 (m, 81 2H), 3.36 (s, 6H), 3.56- 3.60 (m, 1H), 3.62 (s, 3H), 4.01-4.04 (m, 2H), 7.49 (d, J: 4.4 Hz, 1H), 7.67 (d, J = 10.0 Hz, 1H), 7.75-7.78 Prepared by the pmcedure of Exampk: 1 (m, 1H), 8.43 (s, 2H).
H NMR (400 MHz, : 5 1.56-1.62 (m, 2H), 1.91—1.94 (m, 2H), 2.25 (s, 6H), 2.31—2.37 (m, 1H), 2.41 (s, 3H), 2.87- 2.93 (m, 2H), 3.47 (s, 3H), 3.72-3.76 (m, 2H), 7.10 (dd, .1: 1.2, 8.0 Hz, 1H), 7.15 (d, .1: 8.0 Hz, 1H), 7.28-7.31 (m, 1H), 7.49— 7.52 (m, 2H), 7.80 (d, J: 2.0 Hz, 1H).
H NMR (400 MHz, : 5 1.92—2.04 (m, 2H), 2.24-2.26 (m, 2H), 2.79 (s, 3H), 3.14—3.20 (m, 2H), 3.30 (s, 6H), 3.37— 3.40 (m, 1H), 3.61 83 (s, 3H), 3.94—3.98 (m, 2H), 7.16 (d, .1: 9.6 Hz, 1H), 7.42 (d, .1 = 8.0 Hz, 1H), 7.61 (d, .1: .0 HZ, 1H), 7.71-7.76 (m, 2H),7.84 (d, .1: 1.2 Hz, 1H .
H NMR (400 MHz, CDC13):51.61-1.69(m, 2H), 1.98-2.01 (m, 2H), 2.32 (s, 6H), 2.32—2.33 (m, 1H), 2.88-2.94 (m, 2H), 84 (s, 3H), 3.65-3.69 (m, 2H), 6.73 (dd, J: 1.2, 8.8 Hz, 1H), 7.00 (dd, J=1.2, 8.0 HZ, 1H), 7.19-7.30 (m, 2H), 7.37 (s, 1H), 7.52 (d, .1: 8.4 Hz, 1H), 7.90 (s, 1H), 10.65 (br, 1H).
H NMR (400 MHz, CD3OD): 5 .92 (m, 2H), 2.15-2.18 (m, 2H), 3.12-3.18 (m, 2H), 3.40- 3.46 (m, 1H), 3.85-3.88 85 (m, 5H), 6.81 (d, .1: 8.4 Hz, 1H), 6.98-7.05 (111,2 H), 7.25 (dd, .1: 1.2, 8.4 Hz, 1H), 7.42 (d, J: 11.2 Prepared 333/1706 pmcedure oi‘Emnmie 1 HZ, 1H), 7.61 (t, J = 7.2 Hz, 1H .
H NMR (400 MHz, CDgOD): 8 .92 (m, 2H), 2.14-2.17 (m, 2H), 3.11-3.17 (m, 2H), 3.42- 3.47 (m, 1H), 3.59 (s, 3H), 3.85-3.88 (m, 2H), 6.81 (d, J: 8.4 Hz, 1H), 6.98-7.05 (m, 2 H), 7.25 (dd, J: 1.2, 8.0 Hz, 1H), 7.42 (d, J: .4 Hz, 1H), 7.60 (dd, J: 0.8, 7.6 Hz, 1H).
H NMR (400 MHz, DMSO-d6): 8 ppm 1.79 (br. s., 2 H) 2.11 (br. s., 2 H) 2.97 (br. s., 2 H) 3.10 - 3.31 (m, 1 H) 3.46 (br. s., 3 87 H) 3.74 (d, J=18.19 Hz, 2 H) 4.03 (br. s., 3 H) 7.12 (d, J=13.39 Hz, 1 H) 7.40 - 7.61 (m, 4 H) 7.71 (br. s., 1 H) 7.87 - 8.07 (m, 1 H) 9.15 (br. s., 2 H).
H NMR (400 MHz, METHANOL-d4): 8 ppm 1.84 (d, J=13.39 Hz, 2 H) 2.11 (d, J=13.14 Hz, 2 H) 3.05 - 3.17 (m, 2 H) 3.35 - 3.40 (m, 1 H) 3.59 (s, 3 H) 3.83 (d, J=14.40 Hz, 2 H) 7.17 (d, J=8.08 Hz, 2 H) Prepared by the pramedur? 0f EixampEe 1 7.41 (d, 1 Hz, 1 H) 7.44 - 7.52 (m, 2 H) 7.57 (s, 1 H) 7.98 (s, 1 H) 8.54 (br. s., 1 H).
H NMR (400 MHz, METHANOL-d4): 8 ppm 1.74 - 1.96 (m, 2 H) 2.11 (d, J=12.13 Hz, 2 H) 3.08 (q, J=11.54 Hz, 2 H) 3.38 (br. s., 1 H) 3.57 (br. s., 3 H) 3.71 - 3.93 (m, 2 H) 6.64 (d, J=8.08 Hz, 2 H) Prepared by 11163 procedure 61’ Exampic 1 6.86 (d, J=8.08 Hz, 2 H) 7.07 - 7.17 (m, 1 H) 7.18 - 7.30 (m, 1 H) 7.31 - 7.43 m, 1 H .
H NMR (400 MHz, METHANOL-d4): 8 ppm 1.72 - 1.93 (m, 2 H) 2.09 (d, J=11.62 Hz, 2 H) 2.75 (s, 3 H) 2.99 - 3.14 (m, 2 H) 3.36 - 3.43 (m, 1 H) 3.56 (s, 3 H) 3.78 (d, J=12.38 Hz, 2 H) 6.54 (d, J=7.83 Hz, 2 H) 6.89 (d, Prepared by the procedure 0fExamp1e 1 J=7.83 Hz, 2 H) 7.27 (d, J=8.34 Hz, 1 H) 7.32 - 7.43 (m, 1 H) 7.48 - 7.62 (m, 1 H NMR (400 MHz, METHANOL-d4): 8 ppm 1.86 (d, J=11.87 Hz, 2 H) 2.12 (d, J=11.12 Hz, 2 H) 2.96 (s, 3 H) 3.11 (t, J=12.25 Hz, 2 H) 3.40 (br. s., 1 H) 3.57 (s, 3 H) 3.84 (d, J=12.38 Hz, 2 H) 6.90 (d, J=8.59 Hz, 1 H) 7.05 (t, Prepared by ?le ure 9f c 1 J=8.46 Hz, 1 H) 7.12 (d, J=12.38 Hz, 1 H) 7.26 (d, J=8.34 Hz, 1 H) 7.40 (d, J=10.61 Hz, 1 H) 7.60 (t, J=7.20 Hz, 1 H).
H NMR (400 MHz, CHLOROFORM-d): 8 ppm 1.71 (m, J=11.37 Hz, 2 H) 1.74 (br. s., 1 H) 2.04 (d, J=11.87 Hz, 2 H) 2.38 (br. s., 6 H) 2.96 (t, J=12.76 Hz, 2 H) 3.55 (s, 3 H) 3.71 (d, J=12.88 Hz, 2 Prepared 135/1106 pmcedure ofExmane 1 H) 3.91 (s, 3 H) 6.73 (d, J=8.59 Hz, 1 H) 7.12 (d, J=7.83 Hz, 1 H) 7.34 (d, J=10.11Hz, 1 H) 7.43 (t, J=7.07 Hz, 1 H) 7.53 (d, J=8.34 Hz, 1 H) 7.81 (br. s., 1 H).
H NMR (400 MHz, DMSO-d6): 8 ppm 1.32 (td, J=7.01, 1.39 Hz, 3 H) 1.58 (d, 2 Hz, 2 H) 1.92 (d, J=11.62 Hz, 2 H) 2.80 (s, 3 H) 2.91 - 3.03 (m, 2 H) 3.08 (br. s., 1 H) 3.69 (d, J=10.36 Hz, 2 H) 4.29 - 4.40 (m, 2 H) 6.86 (s, 1 H) Prepareci by the pra‘medur? 0f ne 1 6.89 (d, J=8.08 Hz, 1 H) 7.21 (d, J=8.08 Hz, 1 H) 7.56 (d, J=1.77 Hz, 1 H) 7.81 - 7.86 (m, 1 H) 8.33 (s, 3 H).
H NMR (400 MHz, DMSO-d6) 8 ppm 1.28 (td, J=7.01, 2.40 Hz, 3 H) 1.58 (br. s., 2 H) 1.89 (br. s., 2 H) 2.92 - 3.02 (m, 2 H) 3.07 (br. s., 1 H) 3.43 (s, 3 H) 3.68 (d, J=13.39 Hz, 2 H) 4.21 - 4.29 (m, 2 H) Prapared by the pmcedure (H‘Elxampie 1 6.73 (d, J=3.79 Hz, 1 H) 6.80 (s, 1 H) 6.93 (d, J=7.83 Hz, 1 H) 7.20 (d, J=8.59 Hz, 1 H) 7.54 (d, J=8.08 Hz, 1 H) 7.80 - 7.85 m,1H 8.31 s,3H.
H NMR (400 MHz, DMSO-d6): 8 ppm 1.31 (t, J=6.69 Hz, 3 H) 1.73 (d, J=9.09 Hz, 2 H) 2.00 (d, J=12.13 Hz, 2 H) 2.99 (t, J=12.51 Hz, 2 H) 3.28 (br. s., 1 H) 3.43 (s, 3 H) 3.71 (d, J=12.38 Hz, 2 H) 3.95 - Prepared by the pi‘?cedure of Example 1 4.06 (m, 2 H) 6.83 (d, J=8.08 Hz, 2 H) 7.01 (d, J=8.59 Hz, 2 H) 7.18 (d, J=8.59 Hz, 1 H) 7.41 (d, J=10.86 Hz, 1 H) 7.61 (m, 1 H) 7.79 (t, J=7.83 Hz, 1 H 8.07 br. s., 3 H.
H NMR (400 MHz, DMSO-d6) 8 ppm 1.74 (d, J=10.36 Hz, 2 H) 2.00 (d, J=11.62 Hz, 2 H) 2.99 (t, J=12.25 Hz, 2 H) 3.43 (s, 3 H) 3.64 (br. s., 2 H) 3.71 (d, 7 Hz, 2 H) 4.07 (br. s., 2 H) 6.85 (d, J=8.34 Prepareci by the pi‘medur? 0f Eixan’q?e 1 Hz, 2 H) 7.01 (d, J=8.34 Hz, 2 H) 7.18 (d, J=8.08 Hz, 1 H) 7.41 (d, J=10.36 Hz, 1 H) 7.58 - 7.67 (m, 1 H) 7.79 (t, J=7.45 Hz, 1 H) 8.14 (br. s., 3 H).
H NMR (400 MHz, METHANOL-d4): 8 ppm 1.87 (d, J=11.12 Hz, 2 H) 2.13 (d, J=12.13 Hz, 2 H) 3.04 - 3.21 (m, 2 H) 3.38 (d,J=10.61Hz, 1 H) 3.57 (s, 3 H) 3.77 - 3.88 (m, 4 Prapared by the pmcedure (H‘Elxampie 1 H) 4.02 (br. s., 2 H) 6.86 (d, J=7.83 Hz, 2 H) 7.04 (d, J=8.34 Hz, 2 H) 7.47 - 7.53 (m, 2 H) 7.57 (d, J=7.58 Hz, 2 H .
H NMR (400 MHz, DMSO-d6) 8 ppm: 1.74 (d, J=11.87 Hz, 2 H) 2.00 (d, J=12.38 Hz, 2 H) 2.99 (t, J=12.25 Hz, 2 H) 3.28 (br. s., 1 H) 3.43 (s, 3 H) 3.44 - 3.54 (m, 2 H) 3.70 (m, 5 H) 3.90 - 4.05 (m, 2 H) 6.85 (d, J=8.34 Hz, 2 H) 7.01 Prepared by the pmcedure ofExampic 1 (d, J=7.83 Hz, 2 H) 7.18 (d, J=8.08 Hz, 1 H) 7.42 (d,J=10.61Hz, 1 H) 7.79 (t, J=7.20 Hz, 1 H) 8.11 (br. s., 3 H).
H NMR (400 MHz, DMSO-d6): 8 ppm 1.73 (d, J=11.37 Hz, 2 H) 2.00 (d, J=12.63 Hz, 2 H) 2.64 - 2.76 (m, 2 H) 3.00 (t, J=12.13 Hz, 2 H) 3.29 (br. s., 1 H) 3.43 (br. s., 3 H) 3.48 (d, J=9.85 Hz, 2 H) ed? by the pra‘medur? 0f EixampEe 1 3.69 - 3.77 (m, 2 H) 7.00 (d, J=7.33 Hz, 2 H) 7.13 (d, J=7.83 Hz, 2 H) 7.18 (d, J=8.34 Hz, 1 H) 7.34 - 7.42 (m, 1 H) 7.75 - 7.80 (m, 1 H) 8.06 (br. s., 3 H).
H NMR (400 MHz, DMSO-d6): ppm 1.73 (d, J=12.13 Hz, 2 H) 1.94 - 2.03 (m, 2 H) 3.00 (br. s., 2 H) 3.29 (br. s., 1 H) 3.44 (s, 3 H) 3.48 (d, J=9.60 Hz, 2 H) 3.70 (br. s., 2 H) 7.06 (d, J=7.33 Hz, 2 H) 7.16 - 7.25 (m, 3 H) 7.41 (d, J=10.86 Hz, 1 H) 7.75 Prepared by the procedure 61" Exampie 1 - 7.82 (m, 1 H) 8.05 (br. s., 3 H NMR (400 MHz, METHANOL-d4): 8 ppm 1.78 - 1.94 (m, 2 H) 2.13 (d, J=11.87 Hz, 2 H) 3.10 (t, 1 Hz, 2 H) 3.39 (d, J=12.13 Hz, 1 H) 3.57 (s, 3 H) 3.73 - 3.93 (m, 2 H) 6.99 - 7.09 (m, 2 H) 7.12 - 7.25 (m, 3 H) 7.37 Prepared by the prmtedure ofExmane 1 (d, J=10.36 Hz, 1 H) 7.57 (t, J=6.95 Hz, 1 H).
H NMR (400 MHz, METHANOL-d4): 8 ppm 1.79 - 1.93 (m, 2 H) 2.13 (d, J=12.38 Hz, 2 H) 3.11 (t, J=12.63 Hz, 2 H) 3.39 (d, 2 Hz, 1 H) 3.57 102 (s, 3 H) 3.85 (d, J=13.64 Hz, 2 H) 6.89 (d, J=7.83 Hz, 1 H) 6.96 - 7.07 (m, 2 H) 7.23 (d, J=8.34 Hz, 1 Prepared by the pmcedurf: ofExamp1c 1 H) 7.25 - 7.33 (m, 1 H) 7.38 (d, J=10.36 Hz, 1 H) 7.58 (t, J=7.20 Hz, 1 H).
H NMR (400 MHz, METHANOL-d4): 8 ppm 1.77 - 1.95 (m, 2 H) 2.13 (d, J=11.62 Hz, 2 H) 3.12 (t, J=12.76 Hz, 2 H) 3.36 - 3.45 (m, 1 H) 3.55 (s, 3 H) 103 3.86 (d, J=13.64 Hz, 2 H) 6.78 (d, J=6.57 Hz, 2 H) 6.89 (t, J=9.22 Hz, 1 H) 7.24 (d, J=8.08 Hz, 1 H) 7.43 (d, J=10.11 Hz, 1 H) 7.62 (t, J=7.07 Hz, 1 H).
H NMR (400 MHz, METHANOL-d4): 8 ppm 1.79 - 1.93 (m, 2 H) 2.12 (d, J=11.62 Hz, 2 H) 3.11 (t, J=12.63 Hz, 2 H) 3.33 - 104 3.49 (m, 1 H) 3.57 (s, 3 H) 3.85 (d, J=13.64 Hz, 2 H) 6.87 (br. s., 1 H) 7.11 - 7.25 (m, 3 H) 7.42 (d, J=10.36 Hz, 1 H) 7.60 (t, J=7.20 Hz, 1 H).
H NMR (400 MHz, METHANOL-d4): 8 ppm 1.79 - 1.92 (m, 2 H) 2.12 (d, J=11.62 Hz, 2 H) 3.05 - 3.29 (m, 5 H) 3.40 (br. s., 1 H) 3.58 (s, 3 H) 3.80 - 3.94 (m, 2 H) 7.16 (d, J=7.58 Hz, 1 H) 7.36 - 7.48 (m, 3 H) 7.58 (t, J=7.20 Hz, 1 H) Prepared by 11163 procedure 91’ Exampic 1 7.87 (d, J=8.08 Hz, 2 H).
H NMR (400 MHz, CHLOROFORM-d): 8 ppm 1.89 (d, J=11.12 Hz, 2 H) 2.16 (d, J=10.86 Hz, 2 H) 3.05 (t, J=11.87 Hz, 2 s., 1 H) 3.55 106 H) 3.28 (br. (s, 3 H) 3.71 (d, J=12.13 Hz, 2 H) 7.04 (d, J=8.34 Hz, 1 H) 7.10 (d, J=8.08 Hz, 2 H) 7.27 - 7.30 (m, 1 Prepared by 11163 procedure 91’ Example 1 H) 7.33 - 7.44 (m, 2 H) 8.31 (br.s., 1H).
H NMR (400 MHz, METHANOL-d4): 8 ppm 1.81 - 1.94 (m, 2 H) 2.13 (d, J=12.13 Hz, 2 H) 3.12 (t, J=12.38 Hz, 2 H) 3.36 (s, 3 H) 3.41 (br. s., 1 H) 3.58 (s, 3 H) 3.84 (d, J=12.63 Hz, 2 H) 4.45 (s, 2 H) 7.14 (d, J=7.58 Hz, 2 H) 7.23 (d, J=7.83 Hz, 1 Prapared by the pmcedure (H‘Elxampie 1 H) 7.28 (d, J=7.83 Hz, 2 H) 7.34 (d, J=10.61 Hz, 1 H) 7.55 (t, J=7.20 Hz, 1 H NMR (400 MHz, DMSO-d6): 8 ppm 1.62 - 1.78 (m, 2 H) 2.00 (d, J=11.87 Hz, 2 H) 3.02 (t, J=12.00 Hz, 2 H) 3.32 (s, 3 H) 3.76 (d, J=12.88 Hz, 2 H) 6.87 (s, 1 H) 7.95 (br. s., 3 H) 8.01 - 8.08 (m, 1 H) 8.08 - 8.12 (m, 1 H) 8.16 (d, 2 Hz, 1H).
HNMR (400 MHz, CD30D)I 8 0.37-0.39 (m, 2H), 0.60-0.65 (m, 2H), 1.29-1.32 (m, 1H), 1.59- 1.64 (m, 2H), 2.10-2.14 109 (m, 2H), .14 (m, 2H), 3.43-3.47 (m, 1H), 6.81 (d, J: 7.2 Hz, 2H), 4.92-4.95 (m, 2 H), 6.66 (s, Prepared by the pmcedul‘e pkz 13 1H), 7.84-7.88 (m, 1H), 7.99-8.05 (m, 2H).
H NMR (400 MHz, CD30D)I 8 1.40-1.41 (m, NH2 2H), 1.81-1.84 (m, 2H), 2.75-2.78 (m,1H), 2.89- 2.95 (m, 2H), 3.37 (s, 3H), 110 352 3.65-3.68 (m, 2H), 3.77 (s, 1H), 7.66 (t, J: 8.0 Hz, 1H), 7.89 (d, J: 8.0 Hz, 314 1H), 7.96 (d, J: 8.4 Hz, H NMR (400 MHz, DMSO-d6): 1.75-1.83 (m, 2H), 2.06 ( d, J: 10.8 Hz, 2H), 2.99 (t, J: 11.6 Hz, 2H), .30 (m, 1H), 111 442 3.42 (s, 3H), 3.68-3.74 (m, 5H), 6.85 (d, J: 8.0 Hz, 2H), 7.02-7.08 (m, 3H), 7.28-7.45 (m, 2H), 8.38- 8.44 (m, 2H).
H NMR (400 MHz, CDgOD): 1.09-1.17 (m, 2H), 1.57-1.62 (m, 2H), .56 (m, 3H), 2.96- 112 429 3.03 (m, 2H), 3.35 (s, 3H), 3.78 (s, 3H), 6.02 (s, 1H), 6.37 (s, 1H), 6.79-6.97 (m, P1613211661 by 11163 procedure 91' 5x31311311: 1 3H), 7.13-7.27 (m, 2H), 7.42-7.52 (m, 2H).
H NMR (400 MHz, CDgOD): 1.06-1.17 (m, 2H), 1.57-1.62 (m, 2H), 2.49-2.56 (m, 3H), 2.96- 3.09 (m, 2H), 3.36 (s, 3H), 113 431 3.78 (s, 3H), 6.07 (s, 1H), 6.40 (s, 1H), 6.97-7.20 (m, 4H), 7.27 (d, J: 11.8 Hz e1 ,1H), 7.45 (s, 1H), 7.61- 7.66 (m, 1H).
H NMR (400 MHz, CDgOD): 8 152-155 (m, 2H), 1.87-1.90 (m, 2H), 2.83-3.95 (m, 3H), 3.49 (s, 114 414 3H), 3.59-3.67 (m, 5H), 6.23 (s, 1H), 6.81 (d,.]= 8.4 Hz, 1H), 6.98-7.08 (m, P’repareci by the pra‘medma 0f Eixan’q?e 1 4H), 8.14 (t, J: 8.0 Hz, 1H 7.24 , m, 3H.
H NMR (400 MHz, CD30D): 5 .05 (m, 2H), 2.16-2.19 (m, 2H), 3.12—3.20 (m, 2H), 3.44— 3.49 (m, 1H), 3.62 (s, 3H), 115 410 3.88 (s, 3H), 3.90—3.92 (m, 2H), 6.88—6.90 (m, 1H), 7.02-7.06 (m, 1H), 7.08— 7.13 (m, 1H), 8.07 (d, .1: 6.0 Hz, 2H), 8.79 (d, .1: 6.0 Hz, 2H).
H NMR (400 MHz, CD30D): 5 1.50—1.53 (m, 2H), 1.86-1.89 (m, 2H), 2.82-3.95 (m, 3H), 3.48 (s, 116 415 3H), 3.60—3.69 (m, 5H), 6.24 (s, 1H), 6.81 (d, .1: 8.4 Hz, 1H), 7.03 (s, 1H), 7.18—7.24 (m 4H), 8.17 (t, .1 = 4.4 Hz, 1H).
H NMR (400 MHz, CD30D): 5 1.62—1.68 (m, 2H), .03 (m, 2H), 2.96—3.06 (m, 3H), 3.55 (s, 117 444 3H), 3.71—3.74 (m, 5H), 3.81 (s, 3H), 6.36 (d, .1: 3.2 Hz, 1H), 6.65 (d, .1: 8.8 Hz, 1H), 6.93 (t, .1: 8.8 Hz, 1H), 7.13 (d,.]= 3.2 Hz, 1H),7.29-7.38 (m, 4H).
H NMR (300 MHz, CD30D): 5 1.84—1.89 (m, 2H), 2.12-2.16 (m, 2H), 3.13 (t, .1: 12.0 Hz, 2H), .41(rn, 1H), 3.57 (s, 118 434 3H), 3.84 (s, 3H), 3.84— 3.86 (m, 2H), 6.79—6.82 (m, 1H), 6.93—7.00 (m, 2 H), 7.38 (t, .1: 7.5 Hz, 1H), 7.54 (d, .1: 8.4 Hz, Prepareci by the pi‘medur? 0f Eixan’q?e 1 1H), 7.64 (d, .1: 7.8 Hz, 1H), 7.78 (s, 1H).
H NMR (300 MHz, DMSO-d6): 8 1.70-1.74 (m, 2H), 1.99-2.03 (m, 2H), 2.95 (t, J: 12.0 Hz, 2H), 3.23-3.24 (m, 1H), 3.44 (s, 3H), 3.74 (s, 3H), 3.84-3.86 (m, 2H), 6.68- 6.70 (m, 1H), 6.91-6.96 (m, 2 H), 7.31-7.34 (m, Prepared by the ure of Example 1 1H), 7.40-7.59 (m, 2H), 7.77-7.80 (m, 1H), 8.34 (m, 3H).
Preparation 120A: [1-(5-chlorocyanomethyloxo-1,6-dihydro-pyrimidinyl)- piperidinyl]-carbamic acid tert-butyl ester A mixture ofN—[1-(5,6-dichloromethyloxo(3-hydropyrimidinyl))(4- dyl)](tert—butoxy)carboxamide (2.4 g, 6.38 mmol), Zn(CN)2 (388 mg, 3.32 mmol) and Pd(PPh3)4 (740 mg, 0.64 mmol) in DMF (20 mL) was stirred at 130 0C for 5 h under N2 here. The reaction mixture was cooled to RT and ?ltered. The ?ltrate was concentrated in vacuo, and the residue was puri?ed by preperative HPLC to give 200 mg ofthe title product (9%). [M+H] Calc’d for C16H22C1N503, 368; Found, 368.
Preparation 120B: {1-[4-cyano(3-?uoromethoxy-phenyl)methyloxo-1,6- dihydro-pyrimidinyl]-piperidinyl} -carbamic acid tert-butyl ester F110c A mixture of chlorocyanomethyloxo-1,6-dihydro-pyrimidin yl)-piperidinyl]-carbamic acid tert-butyl ester (200 mg, 0.54 mmol), 3-?uoro methoxybenzeneboronic acid (278 mg, 1.63 mmol), Pd(dppf)2Clz (119 mg, 0.16 mmol), and Na2C03 (173 mg, 1.63 mmol) in dioxane (5 mL) and H20 (1 mL) was degassed with N2 and stirred at 145 0C in the microwave for 2 h. The reaction e was cooled to RT and ?ltered. The ?ltrate was concentrated in vacuo and the residue puri?ed by ative HPLC to to give 110 mg ofthe desired t (45%). [M+H] Calc’d for ngHngN504, 458; Found, 458.
Example 120: 2-(4-amino-piperidinyl)-5 -(3-?uoromethoxy-phenyl)methyl oxo- 1 ,6-dihydro-pyrimidinecarbonitrile A mixture of {1-[4-cyano(3-?uoromethoxy-phenyl)methyloxo-1,6- dihydro-pyrimidinyl]-piperidinyl} -carbamic acid tert-butyl ester (100 mg, 0.23 mmol) in EA (5 mL) was added a 5N HCl solution in EA (5 mL) was stirred at RT for 2 h. The solvent was concentrated in vacuo to give 85 mg of the title product as the HCl salt (93 %). 1H NMR (400 MHz, CD30D)C 8 1.71-1.75 (m, 2H), 1.89-2.03 (m, 2H), 2.96- 3.02 (m, 2H), 3.27-3.31 (m, 1H), 3.42 (s, 3H), 3.69-3.73 (m, 2H), 3.83 (s, 3H), 7.06 (t, J = 8.0 Hz, 1H), 7.17-2.01 (m, 2H). [M+H] Calc’d for ClgHzoFNsoz, 358; Found, 358.
Preparation 121A: {1-[5-cyano(4-cyano?uoro-phenyl)methyloxo-1,6- dihydro-pyrimidinyl]-piperidinyl} -carbamic acid tert-butyl ester A mixture of chloro(4-cyano?uoro-phenyl)methyloxo-1,6- dihydro-pyrimidinyl]-piperidinyl} -carbamic acid tert-butyl ester (460 mg, 1 mmol), Zn(CN)2 (175 mg 1.5 mmol) and Pd(PPh3)4(116 mg, 0.0.1 mmol) in DMF (5 mL) was stirred 4 h at 150 0C under N2 atmosphere. The mixture was cooled to RT and ?ltered. The ?ltrate was concentrated in vacuo, and the residue puri?ed by preperative HPLC to give 150 mg ofthe title t as a yellow solid (33%). [M+H] Calc’d for C23H25FN603, 453; Found, 453.
Example 121: 2-(4-amino-piperidinyl)(4-cyano?uoro-phenyl)methyloxo- 1 ,6-dihydro-pyrimidine-5 -carbonitrile To a mixture of {l-[5-cyano(4-cyano?uoro-phenyl)-l-methyloxo-l,6- dihydro-pyrimidinyl]-piperidinyl} -carbamic acid tert—butyl ester (150 mg, 0.33 mmol) in EA (5 mL) was added a 5 N HCl solution in EA (5 mL), and the mixture was stirred at RT for 2 h. The solvent was concentrated in vacuo to give 120 mg the title product as HCl salt (94%). 1H NMR (400 MHZ, CD30D)C 8 1.67-1.72 (m, 2H), 2.02-2.06 (m, 2H), 3.13-3.16 (m, 2H), 3.34-3.38 (m, 1H), 3.42 (s, 3H), .02 (m, 2H), 7.82- 7.90 (m, 3H). [M+H] Calc’d for FN6O, 353; Found, 353.
Preparation 122A: 4-cyano?uoro-benzoyl chloride A mixture of o?uoro-benzoic acid (2.0 g, 12.12 mmol) in SOClz (20 mL) was re?uxed for 2 h, and SOClz was removed in vacuo to give 4-cyano?uoro- benzoyl chloride (2.2 g, 99%). The crude was carried to the next step without ?arther puri?cation.
Preparation 122B: 3-(4-cyano?uoro-phenyl)(4-methoxy-phenyl)oxo-propionic acid methyl ester To a solution of (4-methoxy-phenyl)-acetic acid (2.18 g, 12.12 mmol) in THF (20 mL) was added LiHMDS (18.2 mL, 18.18 mol) at -78°C and the mixture was d for 30 min. A solution of 4-cyano?uoro-benzoyl de (2.2 g, 12 mmol) in THF was added dropwise at -78°C; and the reaction mixture was allowed to warm up to RT and stirred at ght. Aqueous NH4Cl was added and the aqueous was extracted with EA (3x). The combined organics were concentrated in vacuo and the residue was puri?ed by silica column chrmatography (1:5, EA: PE) to give 18 g (45%) ofthe title compound. [M+H] Calc’d for C18H14FNO4, 328; Found, 328. ation 122C: {1-[4-(4-cyano?uoro-phenyl)(4-methoxy-phenyl)oxo- 1 ,6- dihydro-pyrimidinyl]-piperidinyl} -carbamic acid tert—butyl ester A mixture of 3-(4-cyano?uoro-phenyl)(4-methoxy-phenyl)oxo- propionic acid methyl ester (1.8 g, 5.5 mmol), (1-carbamimidoyl-piperidinyl)- carbamic acid tert—butyl ester (2.6 g, 9.2 mmol), DIEA (2.4 g, 18.3 mmol) in toluene (50 mL) was re?uxed overnight. The solvent was concentrated in vacuo. The e was suspended in MeOH and the solids were ?ltered to give 100 mg (4%) ofthe title nd. [M+H] Calc’d for C28H30FN504, 520; Found, 520.
Example 122: 4-[2-(4-amino-piperidinyl)(4-methoxy-phenyl)oxo-1,6-dihydro- pyrimidinyl]?uoro-benzonitrile ] To a solution of {1-[4-(4-cyano?uoro-phenyl)(4-methoxy-phenyl)oxo- 1,6-dihydro-pyrimidinyl]-piperidinyl} -carbamic acid tert—butyl ester (50 mg, 0.096 mmol) in EA (10 mL) was added a 5M HCl solution in EA and the mixture was stirred at RT for 2h. The solvent was removed in vacuo and the residue was puri?ed by preparative HPLC to give 18 mg (40%) of the title compound as the hydrochloride salt. 1H NMR (400 MHz, CD30D)C 8 1.81-1.87 (m, 2H), 2.22-2.25 (m, 2H), 3.34-3.38 (m, 2H), 3.56-3.60 (m, 1H), 3.78 (s, 3H), 4.61-4.64 (m, 2H), 6.86 (d, J: 7.2 Hz, 2H), 7.08 (d, J: 8.4 Hz, 2H), 7.37-7.38 (m, 1H), 7.51-7.53 (m, 1H), 7.74 (s,1H). [M+H] Calc’d for C23H22FN502, 420; Found, 420.
II. Biological Evaluation Example 1a: In Vitro Enzyme Inhibition Assay — LSD-1 This assay determines the ability of a test compound to inhibit LSDl demethylase activity. E.coli expressed ength human LSDl sion number 060341) was purchased from Active Motif (Cat#31334). 1 1 1 The enzymatic assay ofLSDl activity is based on Time Resolved- Fluorescence Resonance Energy Transfer (TR-FRET) detection. The tory properties of nds to LSDl were determined in 384-well plate format under the ing reaction conditions: 0.1- 0.5 nM LSDl, 50 nM H3K4mel-biotin labeled peptide (Anaspec cat # 64355), 2 uM FAD in assay buffer of 50 mM HEPES, pH7.3, 10 mM NaCl, 0.005% Brij35, 0.5 mM TCEP, 0.2 mg/ml BSA. Reaction product was determined quantitatively by TR-FRET after the addition of detection reagent Phycolink Streptavidin-allophycocyanin (Prozyme) and Europium-anti—unmodi?ed histone H3 lysine 4 (H3K4) antibody (PerkinElmer) in the ce ofLSDl inhibitor such as 1.8 mM ofTranylcypromine hydrochloride (2-PCPA) in LANCE detection buffer (PerkinElmer) to ?nal concentration of 125 nM and 0.25 nM respectively.
The assay reaction was performed according to the ing procedure: 2 uL ofthe mixture of 150 nM H3K4mel-biotin labeled peptide with 2 uL of l l-point serial diluted test compound in 3% DMSO were added to each well of plate, followed by the addition of 2 uL of 0.3 nM LSDl and 6 uM ofFAD to initiate the reaction. The reaction mixture was then incubated at room temperature for one hour, and ated by the addition of 6 uL of 1.8 mM 2-PCPA in LANCE detection buffer ning 25 nM Phycolink Streptavidin-allophycocyanin and 0.5 nM Europium-anti—unmodi?ed H3K4 antibody. Enzymatic reaction is terminated within 15 minutes if 0.5 LSDl enzyme is used in the plate. Plates were read by EnVision Multilabel Reader in TR-FRET mode (excitation at 320nm, emission at 615nm and 665nm) after 1 hour incubation at room temperature. A ratio was calculated (665/6l5) for each well and ?tted to determine inhibition constant (ICso).
The y of the compounds disclosed herein to inhibit LSDl activity was quanti?ed and the tive IC50 value was ined. Table 3 provides the IC50 values ofvarious substituted heterocyclic compounds disclosed herein.
TABLE 3 4-(2-(4-aminopiperidin- l -yl) - l -methyloxo-5 -p-tolyl- l ,6- dihydropyrimidinyl)benzonitrile 4- [2-(4-amino-piperidin- l -yl) -5 thoxy—phenyl) - l -methyloxo- l ,6-dihydro-pyrimidinyl] -benzonitrile 1 -[2-(4-amin0-piperidin-1 -y1) -5 -(6-meth0xy-pyridin-3 -methy1 0x0-1,6-dihydr0-pyrimidiny1]-benzonitrile 4-[2-(4-amin0-piperidiny1)methy1(6-methy1-pyridin-3 -y1) 6-dihydr0-pyrimidiny1]-benzonitrile 4-[2-(4-amin0-piperidin-1 -y1) (4-meth0xy-pheny1)methy10x0- 1 ,6-dihydr0-pyrimidiny1] -benzonitrile 4-[2-(4-amin0-piperidin-1 -y1) (4-meth0xy-pheny1)methy10x0- 1,6-dihydr0-pyrimidiny1] ?u0r0-benzonitrile 4-amin0-piperidiny1)-5 -(3 -?u0r0meth0xy-pheny1) methyl0x0-1,6-dihydr0-pyrimidiny1]?u0r0-benzonitrile 1 -amin0-piperidin-1 -y1) -5 th0xy-pyridin-3 -y1)methy1 0x0-1,6-dihydr0-pyrimidiny1]?u0r0-benzonitrile 1 -[2-(4-amin0-piperidin-1 -y1) -5 -(6-meth0xy-pyridin-3 -y1)methy1 0x0-1,6-dihydr0-pyrimidiny1]?u0r0-benzonitrile ,_. O 4-[2-(4-amin0-piperidiny1)-5 -(6-ethy1-pyridin-3 -y1)methy1 0x0-1,6-dihydr0-pyrimidiny1]-benzonitrile ,_. ,_. 2-?u0r0 [5-(4-meth0xy-pheny1)methy1(4-methy1amin0- piperidiny1)0x0- 1,6-dihydr0-pyrimidiny1] -benzonitrile 2-?u0r0 [5 -(3 -?u0r0meth0xy-pheny1)methy1(4- methylamino—piperidiny1)0x0- 1,6-dihydr0-pyrimidiny1]- benzonitrile ,_. U.) 4-[2-(4-amin0-piperidiny1)ethy10x0-1,6-dihydr0-pyrimidin y1] ?uoro—benzonitrile ,_. 4; 4-[2-(4-amin0-piperidin-1 -y1) -5 -cyclopentylethyny1methy10X0- 1,6-dihydr0-pyrimidiny1] ?u0r0-benzonitrile ,_. Ul [2-(4-amin0-piperidiny1)(4-cyan0-3 -pheny1)-5 -(4- methoxy—phenyl)0X0-6H-pyrimidiny1]-acetic acid ._. O\ 2-[2-(4-amin0-piperidiny1)(4-cyan0-3 -?u0r0-pheny1)-5 -(4- methoxy—pheny1)0x0-6H-pyrimidiny1]-acetamide ,_. \] 4-[2-(4-amin0-piperidiny1)(3 xy-pr0py1)0X0-1,6- dihydro—pyrimidiny1] ?u0r0-benzonitrile 00 4-[2-(4-amin0-piperidiny1)benzofurany1methy10x0-1,6- dihydro—pyrimidiny1] ?u0r0-benzonitrile O 2-(4-amin0-piperidiny1)(4-cyan0-3 -?u0r0-pheny1)methy1 0x0- 1 ,6-dihydr0-pyrimidine-5 -carb0nitrile NO 4-[2-(4-amin0piperidin-1 -y1) ch10r0methy10x0pyrimidin y1] ?uorobenzonitrile N._. 2-?u0r0[1-methy1(4-methy1amin0-piperidiny1)(6-methy1- pyridin-3 -y1)0x0-1,6-dihydr0-pyrimidiny1]-benzonitrile NN 4-[2-(2,8—diaza-spir0[4.5]dec-8—y1)(3 -?u0r0meth0xy-pheny1) methyl0x0-1,6-dihydr0-pyrimidiny1]?u0r0-benzonitrile N U.) 4- {2-(4-amin0piperidy1)methy10x0[6-(tri?u0r0methy1) (3 - pyridy1)] hydropyrimidiny1} ?u0r0benzenecarb0nitrile NL 4-[2-(4-amin0piperidyl)methy1(2-methy1(2H-indazoly1)) oxohydropyrimidiny1]benzenecarb0nitrile N Ul 4- [2-((3 R)-3 -amin0piperidy1)-5 -(3 -?u0r0meth0xypheny1) methyl0x0hydr0pyrimidinyl] r0benzenecarb0nitrile 4-[2-(4-amin0piperidy1)-5 -(5 meth0xy(3 -5 ,6- N O\ dihydropyridy1))methy10x0hydr0pyrimidiny1] ?uorobenzenecarbonitrile N \l 4- [2-((3 R)-3 -amin0pyrrolidiny1) -5 -(3 -?u0r0meth0xypheny1) methyl0x0hydr0pyrimidiny1]?u0r0benzenecarb0nitrile N 00 4- [2-((3 S)-3 -amin0-piperidin- 1-y1)-5 -(3 -?u0r0meth0xy-pheny1) methyl0x0-1,6-dihydr0-pyrimidiny1]?u0r0-benzonitrile N0 - [2-((3 S)-3 -amin0-pyrrolidiny1)-5 -(3 -?u0r0meth0xy—pheny1) methyl0x0-1,6-dihydr0-pyrimidiny1]?u0r0-benzonitrile U.) 0 4- [2-((3 R)-3 -amin0piperidy1)(4-meth0xypheny1)methy1 oxohydro pyrimidiny1] ?u0r0benzenecarb0nitrile U.) ,_. 4- [2-((3 S)-3 -amin0-piperidin- 1-y1)-5 -(4-meth0xy-pheny1) methy1 0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile 4-[2-(4-amin0methy1-piperidiny1)-5 -(3 -?u0r0meth0xy— pheny1)methy10X0-1,6-dihydr0-pyrimidiny1]?u0r0- benzonitrile U.) U.) 4- [2-(4-amin0piperidyl)methy1-5 -(1-methy1(1H-indazol-S-y1)) ropyrimidiny1]benzenecarb0nitrile 4-{2-(4-amin0-piperidiny1)methy10X0-5 -[1-(2,2,2 -tri?u0r0- wL - az01y1]— 1,6-dihydr0-pyrimidiny1} ?u0r0- benzonitrile U.) £11 4-[2-(4-amin0-piperidiny1)methy1(1-methy1-1H-indazol-5 - y1)0x0-1,6-dihydr0-pyrimidiny1]?u0r0-benzonitrile U.) C\ 4-{2-(4-amin0-piperidiny1)methy10X0-5 -[1-(2,2,2 -tri?u0r0- ethy1)- 1H-pyraz01y1]— 1,6-dihydr0-pyrimidiny1} -benzonitrile U.) \1 4-[2-(4-amin0piperidyl)methy1(2-methy1(2H-indazoly1)) 0x0hydr0pyrimidiny1] ?u0r0benzenecarb0nitrile U.) 00 4- [2-(4-amin0piperidy1)-5 -(3 ,5 -di?u0r0meth0xypheny1) - 1 l- 6-0X0hydr0pyrimidiny1]benzenecarb0nitrile U.) 0 4- [2-(4-amin0piperidy1)(4-cyan0-3 -?u0r0pheny1)-3 -methy10x0hydr0pyrimidiny1]benzoic acid Lo {4- [2-(4-amin0piperidyl)(4-cyan0pheny1) -3 -methy10X0(3 -hydr0 pyrimidin-S -y1)] ?uorophenyl} hylcarb0xamide L,_. 4-[2-(4-amin0piperidy1)(4-cyan0pheny1)methy10X0(3-hydr0 pyrimidin-S-y1)] ?u0r0benzamide LN 4-[2-(4-amin0-piperidiny1)methy10X0(1-0X0-2,3 -dihydr0- ind01y1)-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile 3 -[2-(4-amin0-piperidiny1)(4-cyan0-3 -?u0r0-pheny1)methy1— 6-0x0-1,6-dihydr0-pyrimidiny1]-benzoic acid 4- {5-(3 -?u0r0meth0xy-pheny1)methy10X0[(3 S)- LL (pyrrolidin-3 -y1methy1)-amin0] - 1,6-dihydr0-pyrimidiny1} - benzonitrile 4- {5-(3 -?u0r0meth0xy-phenyl)methy10x0[(3R)- L£11 (pyrrolidin-3 -y1methy1)-amin0] - 1,6-dihydr0-pyrimidiny1} - benzonitrile LO\ 4-[2-[1,4]diazepany1—5 -(3 -?u0r0meth0xy-phenyl)methy1—6- 6-dihydr0-pyrimidinyl]?u0r0-benzonitrile L\1 2-?u0r0 [5 -(3 -?u0r0meth0xy-pheny1)methy10X0 ziny1-1,6-dihydr0-pyrimidinyl]-benzonitrile L 00 4- [5 -(3 -?u0r0meth0xy-pheny1)methy10x0(piperidin y1amin0)- 1 ,6-dihydr0-pyrimidinyl] -benzonitrile LO 4-[2-(4-amin0-piperidiny1)-2'—dimethy1amin0methy10x0-1,6- dihydro— [5 ,5 ']bipyrimidiny1y1] ?u0r0-benzonitrile -[2-(4-amin0-piperidiny1)(4-cyan0-3 -?u0r0-pheny1)methy1— £11 0 6-0x0-1,6-dihydr0-pyrimidiny1]-pyridinecarb0xylic acid methylamide 2-?u0r0 {5-(4-meth0xy-pheny1)methy10X0[(3 S)- (pyrrolidin-3 -y1methy1)-amin0] - 1,6-dihydr0-pyrimidiny1} - benzonitrile 2-1u0r0 {5-(4-meth0xy-pheny1) methy10X0 [(3 R)- £11 N lidin-3 -y1methy1)-amin0] - 1,6-dihydr0-pyrimidiny1} - benzonitrile £11 U.) 2-?u0r0 [5-(4-meth0xy-pheny1) methy10X0(piperidin y1amin0)- 1 ,6-dihydr0-pyrimidinyl] -benzonitrile 2-?u0r0 [5-(4-meth0xy-pheny1)methy1(methy1-(3 S)- £11 L pyrrolidin-3 -y1methy1—amin0)0x0- 1 ,6-dihydr0-pyrimidiny1] - benzonitrile £11 £11 2-?u0r0 [5-(4-meth0xy-pheny1) methy1(methy1-piperidiny1— amin0)0X0- 1 ,6-dihydr0-pyrimidiny1] -benzonitrile £11 0 2-?u0r0 meth0xy-pheny1)methy1(methy1-pyrrolidin-3 - ylmethyl-amin0)0x0- 1 ,6-dihydr0-pyrimidiny1] -benzonitrile £11 \] 4-[2-(4-amin0-piperidiny1)(6-dimethy1amin0-pyridin-3 -y1) methyl0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile £11 00 2-?u0r0 [5-(6-meth0xy-pyridin-3 -y1)methy1(4-methy1amin0- piperidiny1)0x0- 1,6-dihydr0-pyrimidinyl] -benzonitrile £11 0 4-[2-(4-amin0-piperidin-1 -y1) dimethylamin0-phenyl) methyl- 6-0X0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile 4-[2-(4-amin0-piperidiny1)methy10X0-5 -(6-pyrrolidiny1- pyridin-3 -y1)-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile O\ ._. 4-[2-[1,4]diazepany1-5 -(6-meth0xy-pyridin-3 -y1)methy10x0- 1,6-dihydr0-pyrimidiny1] r0-benzonitrile O\N 4-[2-[1,4]diazepany1-5 -(6-meth0xy-pyridin-3 -y1)methy10x0- 1,6-dihydr0-pyrimidiny1] ?u0r0-benzonitrile O\ U.) 4-[2-[1,4]diazepany1-5 -(6-dimethy1amin0-pyridin-3 -y1)methy1- 6-0X0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile O\4; 4- [2-(3 -amin0-azetidiny1)(4-meth0xy-pheny1)methy10x0- 1,6-dihydr0-pyrimidiny1] ?u0r0-benzonitrile 0UI 2-?u0r0[1-methy1(4-methy1amin0-piperidiny1)-5 -(2-methyl- az01y1)0x0-1,6-dihydr0-pyrimidiny1]-benzonitrile 4-[2-[1,4]diazepany1methy1(2-methy1-2H-indaz01y1) 0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile O\ \] 4-[2-[1,4]diazepany1-5 -(6-dimethy1amin0-pyridin-3 -y1)methy1- 6-0x0-1,6-dihydr0-pyrimidinyl]-benzonitrile 4-[2-(4-amin0-piperidiny1)methy1(6-m0rpholiny1-pyridiny1)0x0- 1 ,6-dihydr0-pyrimidiny1] ?u0r0-benzonitrile 4- [2-(3 -amin0methy1-azetidiny1)(4-meth0xy—pheny1)methyl- 6-0X0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile -?u0r0 meth0xy—pheny1)methy1(3 -methy1amin0methy1- azetidin-l-y1)0x0-1,6-dihydr0-pyrimidinyl]-benzonitn'le \] ,_. 4-dimethylamino—piperidiny1)methy1-5 thy1-2H- indazol-S-y1)0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile \]N 4-[2-(4-dimethylamino—piperidiny1)methy1(1-methy1-1H- indazol-S-y1)0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile \] U.) 4-[2-(4-amin0-piperidiny1)(1H-indol-S-y1)methy10x0-1,6- dihydro—pyrimidiny1] ?u0r0-benzonitrile ‘ -[2-(4-amin0-piperidiny1)methy1(1-methy1-1H-ind01-5 -y1) \1 4; 0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile \] Ul 4-[2-(4-amin0-piperidiny1)(1H-ind01y1)methy10x0-1,6- dihydro—pyrimidiny1] ?u0r0-benzonitrile ‘ -[2-(4-amin0-piperidiny1)methy1(1-methy1-1H-ind01y1) \] O\ 0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile \] \l 4-[2-(4-amin0-piperidiny1)(1H-indazoly1)methy10x0- 1,6-dihydr0-pyrimidiny1] ?u0r0-benzonitrile 4- [2-((4R, 3 S)amin0-3 -?u0r0-piperidiny1)(4-meth0xy- \] 00 pheny1)methy10X0-1,6-dihydr0-pyrimidiny1]?u0r0- itrile 4-[2-((4S, 3R)amin0-3 -piperidiny1)(4-meth0xy- \]O pheny1)methy10X0-1,6-dihydr0-pyrimidiny1]?u0r0- benzonitrile 4-[2-(4-dimethylamino—piperidiny1)methy1-5 -(2-methy1-2H- ly1)0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile 00 ,_. 4-[2'-dimethy1amin0(4-dimethy1amin0-piperidiny1)methy1 0x0- 1 ,6-dihydr0- [5 ,5']bipyrimidiny1y1] ?u0r0-benzonitrile 00N 4-[2-(4-dimethylamino—piperidiny1)methy1(6-methy1-pyridiny1)0x0- 1 ,6-dihydr0-pyrimidiny1] ?u0r0-benzonitrile 4- [5 -(6-dimethy1amin0-pyridin-3 -y1)methy1(4-methy1amin0- 00 U.) piperidiny1)0X0-1,6-dihydr0-pyrimidinyl]?u0r0- benzonitrile oo 4; 4-[2-(4-dimethylamino—piperidiny1)(2H-indazoly1)methyl- 6-0X0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile 4-[2-(4-amin0-piperidiny1)-5 -(3 meth0xy-pheny1) 00 £11 deuteratedmethy10x0- 1 ,6-dihydr0-pyrimidiny1]?u0r0- benzonitrile 4-[2-(4-amin0-piperidiny1)-5 -(3 -?uorodeuteratedmeth0xy— pheny1)methy10X0-1,6-dihydr0-pyrimidiny1]?u0r0- benzonitrile 00 \] 2-?u0r0[1-methy1[4-(methy1amin0)piperidiny1]—5 -(1- methylindazol-S -y1) 0x0pyrimidiny1]benz0nitrile 4-[2-(4-amin0piperidiny1)(1H-indazoly1)methy1 0x0pyrimidiny1] ?u0r0benzonitrile 4- [5 -(4-amin0pheny1)(4-amin0piperidiny1)methy1 0x0pyrimidiny1] ?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)methy1-5 -[4-(methy1amin0)pheny1] imidiny1] ?u0r0benzonitrile O._. 4-[2-(4-amin0piperidin-1 -y1) -5 - [3 -?u0r0(methylamin0)phenyl] methyl0x0pyrimidiny1] ?u0r0benzonitrile 0N 4-[2-[4-(dimethylamin0)piperidiny1]—5-(6-meth0xypyridin-3 -y1) methyl0x0pyrimidiny1] ?u0r0benzonitrile O U.) 4- [2-(4-amin0piperidin(6-eth0xy?u0r0pyridin-3 -y1) methyl0x0pyrimidiny1] ?u0r0benzonitrile O4; 4-[2-(4-amin0piperidiny1)(6-eth0xypyridin-3 -y1)methy1 0x0pyrimidiny1] r0benzonitrile 0 UI 4-[2-(4-amin0piperidiny1)(4-eth0xypheny1)methy1 0x0pyrimidiny1] ?u0r0benzonitrile 4-amin0piperidiny1)-5 - hydr0xyeth0xy)pheny1] methyl0x0pyrimidiny1] ?u0r0benzonitrile O \] 4-[2-(4-amin0piperidiny1)-5 - [4-(2-hydr0xyeth0xy)pheny1] methyl0x0pyrimidiny1]benzonitrile 4-[2-(4-amin0piperidiny1)-5 - [4-(2-meth0xyeth0xy)pheny1]— 1 - methyl0x0pyrimidiny1] ?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)-5 - [4-(2 xyethy1)pheny1]— 1 l- 6-0x0pyrimidiny1]?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)[4-(hydr0xymethy1)pheny1]methy1- yrimidiny1]?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)(4-?u0r0pheny1)methy1 imidiny1] ?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)-5 -(3 -?u0r0pheny1)methy1 0x0pyrimidiny1] ?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)-5 -(3 ,5 -di?u0r0phenyl)methy1 0x0pyrimidiny1] ?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)-5 -(3 ,4-di?u0r0pheny1)methy1 0x0pyrimidiny1] ?u0r0benzonitrile 4-amin0piperidiny1)methy1(4-methylsu1f0ny1pheny1) 0x0pyrimidiny1] ?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)(4-ch10r0pheny1)methy1 0x0pyrimidiny1] ?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)-5 -[4-(meth0xymethy1)phenyl] methyl- 6-0x0pyrimidiny1]?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)methy10X0pyrimidiny1] ?uorobenzonitrile 4-[2-(4-amin0-piperidiny1)cyclopr0py1methy10X0-1, 6- dihydro—pyrimidiny1] ?u0r0-benzonitrile 4-[2-(4-amin0-piperidiny1)cyclopr0py1methy10X0-1, 6- dihydro—pyrimidiny1] ?u0r0-benzonitrile 2-(4-amin0-piperidiny1)(4-ch10r0-3 -?u0r0-pheny1)-5 -(4- methoxy—pheny1)-3 1-3H-pyrimidin0ne 2-(4-amin0-piperidin-1 -y1) (4-hydr0xy-pheny1) -3 -methy1-5 -(1- methyl- 1H-ind01-5 -y1) -3H-pyrimidin0ne 2-(4-amin0-piperidiny1)(4-?u0r0-phenyl)-3 -methy1(1-methy1- 1H-ind01-5 -y1)-3H-pyrimidin0ne 2-(4-amin0-piperidiny1)-3 -methy1(1-methy1-1H-ind01-5 -y1) pheny1-3H-pyrimidin0ne 2-(4-amin0-piperidiny1)-5 -(3 -?u0r0meth0xy-pheny1) -3 -methyl- 6-pyridiny1-3H-pyrimidin0ne 2-(4-amin0-piperidiny1)-3 -methy1(1-methy1-1H-ind01-5 - pyridiny1-3H-pyrimidin0ne 2-(4-amin0-piperidin-1 -y1) (4-meth0xy-pheny1) -3 -methyl(1- methyl- 1H-ind01-5 -y1) -3H-pyrimidin0ne 3 -[2-(4-amin0piperidiny1)-5 -(3 -?u0r0meth0xypheny1)methyl- 6-0X0pyrimidiny1]benzonitrile -[2-(4-aminopiperidinyl)-5 -(3 -?uoromethoxyphenyl) methyl- 6-oxopyrimidinyl]benzonitrile 2-(4-amino-piperidinyl)-5 -(3 -?uoromethoxy—phenyl) methyl- 6-oxo-1,6-dihydro-pyrimidinecarbonitrile 2-(4-amino-piperidinyl)(4-cyano-3 -?uoro-phenyl)methyl 0x0- 1 ,6-dihydro-pyrimidine-5 -carbonitrile 4-aminopiperidinyl)(4-methoxyphenyl)oxo-1H- pyrimidinyl] ?uorobenzonitrile Note: Biochemical assay IC50 data are designated within the following ranges: A: S 0.10 uM B: > 0.10 uM to S 1.0 uM C: > 1.0 tho S 10 uM D: > 10 uM Example 2: In Vitro Enzyme Inhibition Assay — MAO selectivity Human recombinant monoamine oxidase proteins MAO-A and MAO-B are obtained. MAOs catalyze the oxidative deamination ary, secondary and tertiary amines. In order to monitor MAO enzymatic actiVities and/or their inhibition rate by inhibitor(s) of interest, a ?uorescent-based (inhibitor)-screening assay is performed. 3-(2- Aminophenyl)oxopropanamine (kynuramine dihydrobromide, Sigma Aldrich), a non- ?uorescent compound is chosen as a substrate. Kynuramine is a non-speci?c substrate for both MAOs actiVities. While undergoing ive deamination by MAO actiVities, kynuramine is converted into 4-hydroxyquinoline (4-HQ), a ing ?uorescent The monoamine oxidase actiVity was estimated by measuring the conversion of kynuramine into 4-hydroxyquinoline. Assays were conducted in 96-well black plates with clear bottom g) in a ?nal volume of 100 ul. The assay buffer was 100 mM HEPES, pH 7.5. Each experiment was performed in cate within the same ment.
Brie?y, a ?xed amount ofMAO (0.25 ug for MAO-A and 0.5 ug for AO-B) was ted on ice for 15 minutes in the reaction buffer, in the absence and/or in the presence ous concentrations of compounds as disclosed herein (e.g., from 0 to 50 uM, depending on the inhibitor strength). Tranylcypromine (Biomol International) was used as a control for inhibition.
A?er leaving the enzyme(s) interacting with the test nd, 60 to 90 uM of kynuramine was added to each reaction for MAO-B and MAO-A assay respectively, and the reaction was le? for 1 hour at 37 CC in the dark. The oxidative deamination of the substrate was stopped by adding 50 ul of2N NaOH. The conversion ofkynuramine to 4- hydroxyquinoline was monitored by ?uorescence (excitation at 320 nm, on at 360 nm) using a microplate reader te 200, Tecan). Arbitrary units were used to measure levels of ?uorescence produced in the absence and/or in the ce of test compound.
The maximum of oxidative deamination activity was obtained by measuring the amount of 4-hydroxyquino line formed from kynuramine deamination in the absence of test compound and corrected for background ?uorescence. The Ki (IC50) of each inhibitor was determined at Vmax/2. Chemical sis examples 1-94, 101-106, 108- 117, and 120-122 were tested in the above described assay and found to have an IC50 greater than 2 micromolar.
Example 3: LSDl CD1 1b cellular assay To e LSDl inhibitor ef?cacy in cells, a CD1 lb ?ow cytometry assay was performed. LSDl inhibition induces CD1 lb expression in THP-l (AML) cells which is ed by ?ow cytometry. THP-1 cells were seeded at 100,000 cells/well in 10% Fetal Bovine Serum containing RPMI 1640 media in a 24 well plate with a ?nal volume of 500 uL per well. LSDl test compounds were serially diluted in DMSO. The dilutions were added to each well accordingly to a ?nal concentration of 0.2% DMSO. The cells were incubated at 37 degrees Celsius in 5% C02 for 4 days. 250 uL of each well was transferred to a well in a 96 well round bottom plate. The plate was centrifuged at 1200 rpm at 4 degrees Celsius in a n Coulter Alegra 6KR centri?lge for 5 minutes.
The media was removed leaving the cells at the bottom ofthe wells. The cells were washed in 100 uL cold HBSS (Hank’s Balanced Salt Solution) plus 2% BSA (Bovine Serum Albumin) solution and centri?Jged at 1200 rpm at 4 degrees Celsius for 5 minutes. The wash was removed. The cells were resuspended in 100 uL HBSS plus 2% BSA ning 1:15 dilution ofAPC conjugated mouse anti-CD1 1b antibody (BD ngen Cat# 555751) and incubated on ice for 25 minutes. The cells were centrifuged and washed two times in 100 ul HBSS plus 2% BSA. A?er the ?nal spin the cells were resuspended in 100 uL HBSS plus 2% BSA containing lug/mL DAPI (4',6- diamidinophenylindole). The cells were then analyzed by ?ow cytometry in a BD ia machine. Cells were analyzed for CD1 1b expression. The percent of CD1 1b expressing cells for each inhibitor concentration was used to determin e an IC50 curve for each compound analyzed.
] Table 4 provides the ar IC50 values of s substituted heterocyclic compounds disclosed herein.
TABLE 4 4-(2-(4-arninopiperidin- l -yl) - l -rnethyloxo-5 -p-tolyl- l ,6- dihydropyrimidinyl)benzonitrile 4- [2-(4-arnino-piperidin- l -yl) -5 -(4-rnethoxy-phenyl) - l -rnethyloxol ,6-dihydro-pyrirnidinyl] -benzonitrile 4- [2-(4-arnino-piperidin- l -yl) -5 -(6-rnethoxy-pyridin-3 -yl)- l -rnethyl 0x0- 1 ,6-dihydro-pyrirnidinyl] -benzonitrile 4- [2-(4-arnino-piperidin- l -yl) - l -rnethyl-5 -(6-rnethyl-pyridin-3 -yl) 0x0- 1 ,6-dihydro-pyrirnidinyl] -benzonitrile 4- [2-(4-arnino-piperidin- l -yl) -5 -(4-rnethoxy-phenyl) - l -rnethyloxol ,6-dihydro-pyrirnidinyl] -benzonitrile [2-(4-arnino-piperidin- l -yl) -5 -(4-rnethoxy-phenyl) - l -rnethyloxo- 1,6-dihydro-pyrirnidinyl] ?uoro-benzonitrile 4- [2-(4-arnino-piperidin- l -yl) -5 -(3 -?uorornethoxy—phenyl) methyl- 6-oxo- l ydro-pyrimidinyl] ?uoro-benzonitrile arnino-piperidin- l -yl) -5 -(6-rnethoxy-pyridin-3 -yl)- l -rnethyl 0x0- 1 ,6-dihydro-pyrimidinyl] ?uoro-benzonitrile [2-(4-arnino-piperidin- l -yl) -5 -(6-rnethoxy-pyridin-3 -yl)- l -rnethyl 0x0- 1 ,6-dihydro-pyrimidinyl] ?uoro-benzonitrile [2-(4-arnino-piperidin- l -yl) -5 -(6-ethyl-pyridin-3 -yl)- l yloxo- ,_. O l ,6-dihydro-pyrirnidinyl] -benzonitrile -,_.,_. 2-?uoro [5-(4-rnethoxy-phenyl) - l -rnethyl(4-rnethylarnino- piperidin- l -yl)oxo- l,6-dihydro-pyrirnidinyl] nitrile -._. N 2-?uoro [5 -(3 -?uorornethoxy-phenyl) - l -rnethyl(4-rnethylarnino- piperidin- l -yl)oxo- l,6-dihydro-pyrirnidinyl] -benzonitrile -1,_. 4; - [2-(4-arnino-piperidin- l -yl) -5 -cyclopentylethynyl- l yloxo- l ,6- dihydro-pyrimidinyl] ?uoro-benzonitrile -,_. Ul [2-(4-arnino-piperidin- l -yl) (4-cyano-3 -?uoro-phenyl) -5 -(4-rnethoxy— phenyl)oxo-6H-pyrirnidin- l -yl] -acetic acid -._. O\ 2- [2-(4-arnino-piperidin- l -yl) (4-cyano-3 -?uoro-phenyl)-5 -(4- methoxy—phenyl)oxo-6H-pyrirnidin- l -yl] -acetarnide -,_. 00 4- [2-(4-arnino-piperidin- l -yl) -5 -benzofuran-5 -yl- l -rnethyloxo- l ,6- o-pyrimidinyl] ?uoro-benzonitrile NO 4-[2-(4-amin0piperidiny1)ch10r0methy10x0pyrimidiny1]- 2-?u0r0benzonitrile NN 4-[2-(2,8—diaza-spir0[4.5]dec-8—y1)(3 -?u0r0meth0xy-pheny1) methyl0x0-1,6-dihydr0-pyrimidiny1]?u0r0-benzonitrile N U.) 4- {2-(4-amin0piperidy1)methy10x0[6-(tri?u0r0methy1) (3 - pyridy1)] hydropyrimidiny1} r0benzenecarb0nitrile NL 4-[2-(4-amin0piperidyl)methy1(2-methy1(2H-indazoly1)) oxohydropyrimidiny1]benzenecarb0nitrile N Ul 4- [2-((3 R)-3 -amin0piperidy1)-5 -(3 -?u0r0meth0xypheny1)- 1 l- 6-0X0hydr0pyrimidiny1] ?u0r0benzenecarb0nitrile N O\ 4- [2-(4-amin0piperidy1)-5 - (5 -?u0r0meth0xy(3 -5 ,6-dihydr0pyridy1)) - 1 10x0hydr0pyrimidiny1]?u0r0benzenecarb0nitrile N \l 4- [2-((3 R)-3 -amin0pyrrolidiny1) -5 -(3 -?u0r0meth0xypheny1) methyl0x0hydr0pyrimidiny1]?u0r0benzenecarb0nitrile N0 4- [2-((3 S)-3 -amin0-pyrrolidiny1)-5 -(3 -?u0r0meth0xy—pheny1) methyl0x0-1,6-dihydr0-pyrimidiny1]?u0r0-benzonitrile U.) 0 4- [2-((3 R)-3 -amin0piperidy1)(4-meth0xypheny1)methy1 oxohydro pyrimidiny1] ?u0r0benzenecarb0nitrile U.) ,_. 4- [2-((3 S)-3 -amin0-piperidin- 1-y1)-5 -(4-meth0xy-pheny1) methy1 0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile 4-[2-(4-amin0methy1-piperidiny1)-5 -(3 -?u0r0meth0xy-pheny1)- 1-methy10X0-1,6-dihydr0-pyrimidiny1]?u0r0-benzonitrile U.) U.) 4- [2-(4-amin0piperidyl)methy1-5 -(1-methy1(1H-indazol-S-y1)) oxohydropyrimidiny1]benzenecarb0nitrile 4-{2-(4-amin0-piperidiny1)methy10X0-5 -[1-(2,2,2 0r0- wL ethy1)- 1H-pyraz01y1]— 1,6-dihydr0-pyrimidiny1} ?u0r0- benzonitrile U.) £11 4-[2-(4-amin0-piperidiny1)methy1(1-methy1-1H-indazol-5 -y1) 0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile U.) C\ 4-{2-(4-amin0-piperidiny1)methy10X0-5 ,2,2 -tri?u0r0- ethy1)- 1H-pyraz01y1]— 1,6-dihydr0-pyrimidiny1} -benzonitrile U.) \1 4-[2-(4-amin0piperidyl)methy1(2-methy1(2H-indazoly1)) 0x0hydr0pyrimidiny1] ?u0r0benzenecarb0nitrile U.) 00 4- [2-(4-amin0piperidy1)-5 - (3 ,5 -di?u0r0meth0xyphenyl)methy1 oxohydropyrimidiny1]benzenecarb0nitrile Lo {4- [2-(4-amin0piperidyl)(4-cyan0pheny1) -3 -methy10X0(3 -hydr0 pyrimidin-S -y1)] ?uorophenyl} -N-methylcarb0xamide L 4-amin0piperidy1)(4-cyan0pheny1)methy10X0(3-hydr0 pyrimidin-S-y1)] ?u0r0benzamide ‘ -[2-(4-amin0-piperidiny1)methy10x0(1-0X0-2,3 -dihydr0-1H- isoindol-S -y1)- 1 ,6-dihydr0-pyrimidiny1] ?u0r0-benzonitrile LL 4- {5 -(3 -?u0r0meth0xy—pheny1)methy10X0[(3 rrolidiny1methy1)-amin0]—1,6-dihydr0-pyrimidiny1}-benzonitrile L£11 4- {5 -(3 -?u0r0meth0xy—pheny1)methy10X0[(3 R)-(pyrrolidiny1methy1)-amin0]—1,6-dihydr0-pyrimidiny1}-benzonitrile LO\ 1,4]diazepany1(3 -?u0r0meth0xy-phenyl)methy1—6-0x0- 1,6-dihydr0-pyrimidiny1] ?u0r0-benzonitrile L\1 2-?u0r0 [5 -(3 -?u0r0meth0xy-phenyl)methy10x0piperazin- 1-y1- 1 ,6-dihydr0-pyrimidiny1] nitrile L 00 4- [5 -(3 -?u0r0meth0xy-pheny1)methy10x0(piperidin y1amin0)- 1 ,6-dihydr0-pyrimidinyl] -benzonitrile LO 4-[2-(4-amin0-piperidiny1)-2'—dimethy1amin0methy10x0-1,6- dihydro— [5 ,5 ']bipyrimidiny1y1] ?u0r0-benzonitrile -[2-(4-amin0-piperidiny1)(4-cyan0-3 -?u0r0-pheny1)methy1—6- £11 0 0x0-1,6-dihydr0-pyrimidin-5 -y1] -pyridinecarb0xylic acid methylamide £11 {-1 2-?u0r0 {5-(4-meth0xy—phenyl)methy10x0[(3 S)-(pyrrolidiny1methy1)-amin0]—1,6-dihydr0-pyrimidiny1}-benzonitrile £11 N -1u0r0 {5 -(4-meth0xy—phenyl)methy10x0[(3R)-(pyrrolidin-3 - ylmethy1)-amin0] - 1 ,6-dihydr0-pyrimidiny1} -benzonitrile 2-?u0r0 [5-(4-meth0xy-pheny1) methy10X0(piperidin 0)- 1 ,6-dihydr0-pyrimidinyl] -benzonitrile £11 L 2-?u0r0 [5-(4-meth0xy—pheny1)methy1(methy1-(3 S)-pyrrolidiny1methy1—amin0)0x0-1,6-dihydr0-pyrimidiny1]-benz0nitrile £11 £11 2-?u0r0 [5-(4-meth0xy-pheny1) methy1(methy1-piperidiny1— amin0)0X0- 1 ,6-dihydr0-pyrimidiny1] -benzonitrile £11 0 2-?u0r0 [5-(4-meth0xy-pheny1)methy1(methy1-pyrrolidin-3 - ylmethyl-amin0)0x0- 1 ,6-dihydr0-pyrimidiny1] nitrile £11 \] 4-[2-(4-amin0-piperidiny1)(6-dimethy1amin0-pyridin-3 -y1) methyl0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile £11 00 2-?u0r0 [5-(6-meth0xy-pyridin-3 -y1)methy1(4-methy1amin0- piperidiny1)0x0- 1,6-dihydr0-pyrimidinyl] -benzonitrile £11 0 4-[2-(4-amin0-piperidiny1)(4-dimethy1amin0-phenyl)methy1—6- 0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile 4-amin0-piperidiny1)methy10X0-5 -(6-pyrrolidiny1- pyridin-3 -y1)-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile O\ 4-[2-[1,4]diazepany1—5 -(6-meth0xy-pyridin-3 -y1)methy1—6-0x0- 1,6-dihydr0-pyrimidiny1] ?u0r0-benzonitrile O\N 4-[2-[1,4]diazepany1—5 -(6-meth0xy-pyridin-3 -methy1—6-0x0- 1,6-dihydr0-pyrimidiny1] ?u0r0-benzonitrile O\ U.) 4-[2-[1,4]diazepany1(6-dimethy1amin0-pyridin-3 -y1)methy1—6- 0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile O\4; 4- [2-(3 -amin0-azetidiny1)(4-meth0xy-pheny1)methy10x0-1,6- dihydro—pyrimidiny1] ?u0r0-benzonitrile 0UI 0[1-methy1(4-methy1amin0-piperidiny1)-5 -(2-methy1-2H- indazol-S-y1)0x0-1,6-dihydr0-pyrimidinyl]-benzonitrile 4-[2-[1,4]diazepany1methy1(2-methy1-2H-indaz01y1)0x0- 1,6-dihydr0-pyrimidiny1] ?u0r0-benzonitrile O\ \] 1,4]diazepany1(6-dimethy1amin0-pyridin-3 -y1)methy1 0x0-1,6-dihydr0-pyrimidinyl]-benzonitrile 4-[2-(4-amin0-piperidiny1)methy1(6-m0rpholiny1-pyridin-3 - y1)0x0-1,6-dihydr0-pyrimidiny1]?u0r0-benzonitrile 4- [2-(3 methy1-azetidiny1)(4-meth0xy—pheny1)methy1 0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile \] O 2-?u0r0 [5-(4-meth0xy—pheny1)methy1(3 -methy1amin0methy1- azetidin-l-y1)0x0-1,6-dihydr0-pyrimidinyl]-benzonitn'le \] ,_. 4-[2-(4-dimethylamino—piperidiny1)methy1-5 -(2-methy1-2H- indazol-S-y1)0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile \]N 4-[2-(4-dimethylamino—piperidiny1)methy1(1-methy1-1H- indazol-S-y1)0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile \] U.) 4-[2-(4-amin0-piperidiny1)(1H-ind01y1)methy10x0-1,6- dihydro—pyrimidiny1] ?u0r0-benzonitrile 4-[2-(4-amin0-piperidiny1)methy1(1-methy1-1H-ind01-5 -y1) 0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile \] Ul 4-[2-(4-amin0-piperidiny1)(1H-ind01y1)methy10x0-1,6- dihydro—pyrimidiny1] ?u0r0-benzonitrile \] O\ 4-[2-(4-amin0-piperidiny1)methy1(1-methy1-1H-ind01y1) 6-dihydr0-pyrimidinyl]?u0r0-benzonitrile \] \l 4-[2-(4-amin0-piperidiny1)(1H-indazoly1)methy10x0-1,6- dihydro—pyrimidiny1] ?u0r0-benzonitrile \] 00 4- [2-((4R, 3 S)amin0-3 - ?uoro—piperidiny1)-5 -(4-meth0xy-pheny1)- 1-methy10X0-1,6-dihydr0-pyrimidiny1]?u0r0-benzonitrile \]O 4- [2-((4S, 3 R)amin0-3 - ?uoro—piperidiny1)-5 -(4-meth0xy-pheny1)- 1-methy10X0-1,6-dihydr0-pyrimidiny1]?u0r0-benzonitrile 4-[2-(4-dimethylamino—piperidiny1)methy1-5 -(2-methy1-2H- indazoly1)0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile 00 4-[2'-dimethy1amin0(4-dimethy1amin0-piperidiny1)methy1 0x0- 1 ,6-dihydr0- [5 ,5']bipyrimidiny1y1] ?u0r0-benzonitrile 00N 4-[2-(4-dimethylamino—piperidiny1)methy1(6-methy1-pyridin-3 - y1)0X0-1,6-dihydr0-pyrimidiny1]?u0r0-benzonitrile 00 U.) 4- [5 methy1amin0-pyridin-3 -y1)methy1(4-methy1amin0- piperidiny1)0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benz0nitrile oo 4; 4-[2-(4-dimethylamino—piperidiny1)(2H-indazoly1)methy1 0x0-1,6-dihydr0-pyrimidinyl]?u0r0-benzonitrile 4-[2-(4-amin0-piperidiny1)-5 -(3 -?u0r0meth0xy-pheny1) 00 £11 deuteratedmethy10x0- 1 ,6-dihydr0-pyrimidiny1]?u0r0- itrile 4-[2-(4-amin0-piperidiny1)-5 -(3 deuteratedmethoxy-phenyl)- 1-methy10X0-1,6-dihydr0-pyrimidiny1]?u0r0-benzonitrile 00 \] 2-?u0r0[1-methy1[4-(methy1amin0)piperidiny1]—5 -(1- methylindazol-S -y1) 0x0pyrimidiny1]benz0nitrile 00 00 4-[2-(4-amin0piperidiny1)(1H-indazoly1)methy1 0x0pyrimidiny1] ?u0r0benzonitrile 4- [5 -(4-amin0pheny1)(4-amin0piperidiny1)methy1 imidiny1] ?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)methy1-5 -[4-(methy1amin0)pheny1] 0x0pyrimidiny1] ?u0r0benzonitrile ,_. 4-[2-(4-amin0piperidin-1 -y1) -5 - [3 -?u0r0(methylamin0)phenyl] methyl0x0pyrimidiny1] ?u0r0benzonitrile 0N 4-(dimethylamin0)piperidiny1]—5-(6-meth0xypyridin-3 -y1) methyl0x0pyrimidiny1] r0benzonitrile O U.) 4- [2-(4-amin0piperidin- 1-y1)-5 -(6-eth0xy?u0r0pyridin-3 -y1) 0x0pyrimidiny1] ?u0r0benzonitrile O4; 4-[2-(4-amin0piperidiny1)(6-eth0xypyridin-3 -y1)methy1 0x0pyrimidiny1] ?u0r0benzonitrile 0 UI 4-[2-(4-amin0piperidiny1)(4-eth0xypheny1)methy1 0x0pyrimidiny1] ?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)-5 - [4-(2-hydr0xyeth0xy)pheny1] methyl- 6-0x0pyrimidiny1]?u0r0benzonitrile O \] 4-[2-(4-amin0piperidiny1)-5 - [4-(2-hydr0xyeth0xy)pheny1] methyl- 6-0X0pyrimidiny1]benzonitrile 4-[2-(4-amin0piperidiny1)-5 - [4-(2-meth0xyeth0xy)pheny1] methyl- 6-0x0pyrimidiny1]?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)-5 - [4-(2-hydr0xyethy1)pheny1]—1-methy1 0x0pyrimidiny1] ?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)[4-(hydr0xymethy1)pheny1] methy1 0x0pyrimidiny1] ?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)(4-?u0r0pheny1)methy1 0x0pyrimidiny1] ?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)-5 -(3 -?u0r0pheny1)methy1 0x0pyrimidiny1] ?u0r0benzonitrile 4-[2-(4-amin0piperidiny1)-5 -(3 ,5 -di?u0r0phenyl)methy1 0x0pyrimidiny1] ?u0r0benzonitrile 4-[2-(4-aminopiperidiny1)-5 -(3 ,4-di?uorophenyl)methy1 oxopyrimidinyl] ?uorobenzonitrile 4-[2-(4-aminopiperidiny1)methy1(4-methy1sulfonylphenyl) oxopyrimidinyl] ?uorobenzonitrile 4-[2-(4-aminopiperidiny1)(4-chloropheny1)methy1 oxopyrimidinyl] ?uorobenzonitrile 4-[2-(4-aminopiperidiny1)[4-(methoxymethyl)phenyl]methy1 imidinyl] ?uorobenzonitrile 4-[2-(4-aminopiperidiny1)methy1oxopyrimidinyl] ?uorobenzonitrile 4- [2-(4-amino-piperidiny1)cyclopropylmethyloxo-1,6-dihydro- pyrimidinyl] ?uoro-benzonitrile 2-(4-amino-piperidiny1)(4-chloro-3 -?uoro-pheny1)(4-methoxy— pheny1)-3 -methy1-3H-pyrimidinone 4-[2-(4-aminopiperidiny1)(4-methoxypheny1)oxo-1H- pyrimidiny1]?uorobenzonitrile Note: Cellular assay IC50 data are designated within the following ranges: A: S 0.10 uM B: > 0.10 uM to S 1.0 uM C: > 1.0 tho S 10 uM D: > 10 uM Example 4: Kasumi-l AML Cell Line Proliferation Assay (Cell-MTS Assay) Colorimetric cellular assay to assess the ability of LSD-1 small molecule inhibitors to effect the eration of the established AML cancer cell line Kasumi-l.
Assay Background The LSD-1 protein has been shown to play a key role in the biology of a variety of cancer types including SCLC and AML. To demonstrate small molecule tion of LSD-1 as a potential anti-cancer therapy, an assay to measure the degree of proliferative tion in an ished cancer cell line ofAML was implemented.
Assay Principle ] This Cell-MTS assay is a 7-day plate based colorimetric assay which quantifies the amount y generated NADH in the presence and absence of test compound.
These NADH levels are used as a proxy for the quantification of cancer cell proliferation.
Assay Method The established cancer cell line -l with a veri?ed p53 mutation were purchased from American Type Culture Collection (ATCC) and routinely passaged according to ATCC published protocols. For routine assay these cells were seeded at a density of 20,000 cells per 96-well. 24 hours after plating, cells received an 11 point dilution of test compound with ?nal concentration ranges from 100 uM to 2.0 nM. Cells are incubated in the presence of compound for 168 hours at 37 oC, 5% C02. At the end ofthis compound incubation period, 80 ul of media is removed and 20 uL of CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay on (Promega) is added.
The cells are incubated until the OD490 is >0.6. IC50 values are calculated using the [DBS XL?t software package and include ound subtracted OD490 values and normalization to DMSO controls.
Table 5 provides the Kasumi-l cellular IC50 values of s substituted heterocyclic compounds disclosed herein.
TABLE 5 4-(2-(4-aminopiperidin- l -yl) - l -methyloxo-5 -p-tolyl- l ,6- dihydropyrimidinyl)benzonitrile 4- [2-(4-amino-piperidin- l -yl) -5 thoxy—pyridin-3 -yl)- l -methyl 0x0- 1 ,6-dihydro-pyrimidinyl] -benzonitrile 4- [2-(4-amino-piperidin- l -yl) - l -methyl-5 -(6-methyl-pyridin-3 -oxol ,6-dihydro-pyrimidinyl] -benzonitrile 4- [2-(4-amino-piperidin- l -yl) -5 -(4-methoxy—phenyl) - l -methyloxol ,6-dihydro-pyrimidinyl] -benzonitrile 4- [2-(4-amino-piperidin- l -yl) -5 -(4-methoxy—phenyl) - l -methyloxo- 1,6-dihydro-pyrimidinyl] ?uoro-benzonitrile 4- [2-(4-amino-piperidin- l -yl) -5 -(3 -?uoromethoxy—phenyl) methyl- 6-oxo- l ,6-dihydro-pyrimidinyl] ?uoro-benzonitrile 4- [2-(4-amino-piperidin- l -yl) -5 -(6-methoxy—pyridin-3 -yl)- l -methyl 0x0- 1 ,6-dihydro-pyrimidinyl] ro-benzonitrile 4- [2-(4-amino-piperidin- l -yl) -5 -(6-methoxy—pyridin-3 -yl)- l -methyl 0x0- 1 ,6-dihydro-pyrimidinyl] ?uoro-benzonitrile N 4-[2-(4-aminopiperidyl)- l -methyl(2-methyl(2H-indazolyl)) oxohydropyrimidinyl]benzenecarbonitrile U.) 4- {2-(4-amino-piperidin- l -yl) - l loxo-5 -[ l -(2,2,2-tri?uoro- - l zolyl] - l ,6-dihydro-pyrimidinyl} ?uoro- 4-[2-(4-amino-piperidinyl)methyl(1-methyl-1H-indazol-5 -yl) oxo-1,6-dihydro-pyrimidinyl]?uoro-benzonitrile 4- amino-piperidinyl)methyloxo-5 ,2,2-tri?uoro- ethyl)-1H-pyrazolyl]—1,6-dihydro-pyrimidinyl}-benzonitrile 2-?uoro[1-methyl(4-methylamino-piperidinyl)-5 -(2-methyl-2H- indazol-S-yl)oxo-1,6-dihydro-pyrimidinyl]-benzonitrile 4-[2-[1,4]diazepanylmethyl(2-methyl-2H-indazolyl)oxo- 1,6-dihydro-pyrimidinyl] ?uoro-benzonitrile 4-dimethylamino-piperidinyl)methyl-5 -(2-methyl-2H- indazol-S-yl)oxo-1,6-dihydro-pyrimidinyl]?uoro-benzonitrile 4-[2-(4-aminopiperidinyl)(1H-indazolyl)methyl oxopyrimidinyl] ?uorobenzonitrile Example 5: In Vivo Xenograph Study — MCF-7 Xenograph Time release pellets containing 0.72 mg 17-[3 Estradiol are subcutaneously implanted into nu/nu mice. MCF-7 cells are grown in RPMI containing 10% PBS at 5% C02, 37 oC. Cells are spun down and re-suspended in 50% RPMI (serum free) and 50% Matrigel at 1X107 cells/mL. MCF-7 cells are subcutaneously injected /animal) on the right ?ank 2-3 days post pellet implantation and tumor volume (length x width2/2) is monitored bi—weekly. When tumors reach an average volume of ~200 mm3 animals are randomized and treatment is started. Animals are treated with vehicle or compound daily for 4 weeks. Tumor volume and body weight are monitored bi-weekly throughout the study. At the conclusion ofthe treatment period, plasma and tumor samples are taken for pharmacokinetic and pharmacodynamic analyses, respectively.
Example 6: In Vivo Xenograph Study — LNCaP Xenograph LNCaP cells with a stable knockdown ofLSDl (shLSDl cells) or control cells (such as shNTC cells) are ated in the dorsal ?ank of nude mice by subcutaneous injection (such as 3 x 106 cells in 100 ul of 50% RPMI 1640/BD Matrigel). Mouse weight and tumor size are ed once per week and tumor volume is estimated using the a (7i/6)(LXW), where L = length oftumor and W = width oftumor. A two sample t-test is performed to determine statistical differences in mean tumor volume between the two groups.
Unmodi?ed LNCaP cells are inoculated by subcutaneous injection into the dorsal ?ank ofnude mice (such as 3 x 106 cells in 100 ul of 50% RPMI 1640/BD Matrigel). A?er three weeks, mice are injected intraperitoneally once per day with water (control), pargyline (0.53 mg or 1.59 mg; 1 or 3 mM ?nal concentration, assuming 70% bioavailability), or XB154 (4 or 20 ug; 1 or 5 uM ?nal concentration, assuming 70% bioavailability) or treated with a test nd (5 mg/kg each week or 10 mg/kg each week). Treatment continues for three weeks, during which time mouse weight and tumor volume are measured as above. shLSDl LNCaP cells or control cells are injected in nude mice as above. After three weeks, mice are treated with 2.6 ug mitomycin C (predicted ?nal concentration of 1 uM assuming 40% bioavailability), olaparib (for example, about 0.5 mg/kg to 25 mg/kg), or vehicle intraperitoneally once per day for three weeks. In other examples, unmodi?ed LNCaP cells are injected in nude mice as above.
A?er three weeks, mice are treated with test compounds, or vehicle as above, plus MMC or ib. Treatment continues for three weeks, during which time mouse weight and tumor volume are measured as above.
] A decrease in tumor volume compared to control in mice injected with shLSDl cells indicates that LSDl inhibition decreases tumor growth in vivo.
Similarly, a decrease in tumor volume compared to control in mice injected with LNCaP cells and treated with a compound disclosed herein indicates that LSDl inhibition decreases tumor grth in vivo. Finally, a decrease in tumor volume in mice injected with LNCaP cells and treated with a compound disclosed herein plus olaparib as ed to mice treated with a compound disclosed herein alone indicates that inhibition of LSDl plus inhibition ofPARP decreases tumor growth in vivo.
The harvested xenogra? tissue is examined for evidence ofLSDl inhibition.
This is ed with Western blots to examine global levels of the 2MK4 and 2MK9 histone marks, expression ofFA/BRCA genes, FANCD2 ubiquitination, and LSDl protein levels in the cases ofthe shRNA cells. A se in one or more of these parameters indicates the effective inhibition ofLSD 1. Additionally, effects on DNA damage repair are assessed with staining for H2AX foci.
III. ation of ceutical Dosage Forms e 1: Oral Tablet A tablet is prepared by mixing 48% by weight of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, 45% by weight of microcrystalline cellulose, 5% by weight of low-substituted ypropyl cellulose, and 2% by weight ofmagnesium stearate. Tablets are prepared by direct compression. The total weight of the compressed tablets is maintained at 250-500 mg.

Claims (29)

1. A compound having the structure of Formula (I), or a pharmaceutically able salt thereof, NC W N Z X Y O (I) wherein, W is C-H, or C-F; X is hydrogen, n, -CN, C2 alkynyl optionally substituted with C5 cycloalkyl, C5 carbocyclyl-C2-alkynyl, C6 aryl optionally substituted with halogen, C1-C2 alkyl optionally substituted with OH, OMe, methoxy, ethoxy, CF3, carboxyl, carboxamide, amine, NMe2, OCD3, OCH2CH2OH, OCH2CH2OMe, or -SO2Me, or monocyclic or bicyclic C3-C8 heteroaryl containing oxygen or nitrogen and optionally substituted with oxo group, NMe2, C1-C2 alkyl optionally substituted with CF3, methoxy, ethoxy, trifluoromethyl, amide, or N-attached C4 heterocyclyl containing nitrogen or oxygen; Y is hydrogen, CD3, C1-C3 alkyl optionally substituted with COOH, ), or OH, or loalkyl(C1-C3)alkyl; Z is N-attached C3-C8 heterocyclyl containing nitrogen and ally substituted with halogen, NHMe, NMe2, amine, methyl optionally substituted with NHMe or NH2, - N(R)-C5 heterocyclyl containing nitrogen and -N(R)-C4 heterocyclyl(C1-C3)alkyl containing nitrogen; and R is hydrogen or C1-C4alkyl.
2. The nd of claim 1, or a ceutically acceptable salt thereof, wherein W is C-H.
3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein W is C-F.
4. The compound of any one of claims 1-3, or a pharmaceutically acceptable salt thereof, wherein X is en.
5. The compound of any one of claims 1-3, or a pharmaceutically acceptable salt thereof, wherein X is halogen.
6. The nd of any one of claims 1-3, or a pharmaceutically acceptable salt thereof, wherein X is C2 alkynyl.
7. The compound of any one of claims 1-3, or a pharmaceutically acceptable salt thereof, wherein X is C5 carbocyclylC2alkynyl.
8. The compound of any one of claims 1 -3, or a pharmaceutically acceptable salt thereof, wherein X is C6 aryl.
9. The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein X is phenyl.
10. The compound o f any one of claims 1-3, or a pharmaceutically acceptable salt thereof, wherein X is clic or bicyclic C3-C8 heteroaryl.
11. The compound of claim 10, or a pharmaceutically acceptable salt thereof, wherein the monocyclic or bicyclic C3-C8 heteroaryl is chosen from pyridinyl, pyrazolyl, or indazolyl.
12. The compound of any one of claims 1 -11, or a pharmaceutically able salt thereof, wherein Z is -N(H)-C4heterocyclyl(C1-C3)alkyl.
13. The compound of any one of claims 1-11, or a pharmaceutically acceptable salt thereof, wherein Z is -N(Me)-C4heterocyclyl(C1-C3)alkyl.
14. The compound of any one of claims 1 -11, or a pharmaceutically acceptable salt thereof, wherein Z is -N(R)-C4heterocyclylC1alkyl.
15. The compound of any one of claims 1 -11, or a pharmaceutically acceptable salt thereof, wherein Z is N-attached C3-C8 heterocyclyl.
16. The compound of claim 15, or a ceutically acceptable salt thereof, wherein the N- attached C3-C8 heterocyclyl is a 4-, 5-, 6-, or 7-membered N-attached heterocyclyl.
17. The compound o f claim 15, or a pharmaceutically able salt f, wherein the N- attached C3-C8 heterocyclyl is a 6-membered N-attached heterocyclyl.
18. The nd of claim 15, or a pharmaceutically acceptable salt thereof, wherein the N- attached C3-C8 heterocyclyl is piperidine.
19. The compound of claim 18, or a pharmaceutically acceptable salt thereof, wherein the piperidine is 4-aminopiperidine.
20. The compound of any one of claims 1-19, or a ceutically able salt thereof, wherein Y is hydrogen.
21. The compound of any one of claims 1 -19, or a pharmaceutically acceptable salt thereof, wherein Y is C1-C3 alkyl.
22. The nd of claim 21, or a pharmaceutically acceptable salt thereof, wherein the C1-C3 alkyl is a methyl group.
23. The compound of claim 1, or a ceutically acceptable salt thereof, having the any one of the following structures of: (i) ; (ii) ; (iii) ; (iv) ; (v) ; or (vi) .
24. The nd of claim 1, or a pharmaceutically acceptable salt thereof, having the following structure of:
25. The compound of claim 1, or a pharmaceutically acceptable salt thereof, having any one of the following structures of: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and or pharmaceutically acceptable salt thereof.
26. A pharmaceutical composition comprising a compound of Formula (I) according to any one of claims 1 to 25, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.
27. An in vitro method of regulating gene transcription in a cell comprising inhibiting specific demethylase 1 activity by exposing the lysine-specific ylase 1 enzyme to a compound of Formula (I) according to any one of claims 1 to 25.
28. Use of a compound of Formula (I) according to any one of claims 1 to 25, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament effective in the ent of cancer.
29. The use of claim 28, wherein the cancer is acute myeloid leukemia (AML), te cancer, and breast cancer. Celgene Quanticel Research, Inc. By the Attorneys for the Applicant SPRUSON & FERGUSON
NZ725914A 2015-04-30 Inhibitors of lysine specific demethylase-1 NZ725914B2 (en)

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US201461987354P 2014-05-01 2014-05-01
PCT/US2015/028635 WO2015168466A1 (en) 2014-05-01 2015-04-30 Inhibitors of lysine specific demethylase-1

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