WO2018132636A1

WO2018132636A1 - [18f]maleimide-based glycogen synthase kinase-3beta ligands for positron emission tomography imaging and radiosynthesis method

Info

Publication number: WO2018132636A1
Application number: PCT/US2018/013446
Authority: WO
Inventors: Ming-Yu NGAI; Katarzyna N. LEE; Kongzhen HU; Neil NASDEV; Stephen J. Haggarty
Original assignee: The Research Foundation For The State University Of New York
Priority date: 2017-01-12
Filing date: 2018-01-12
Publication date: 2018-07-19

Abstract

The present invention provides a compound having the structure: (Formula I), and a method of inhibiting Glycogen synthase kinase-3 β (GSK-3β) in a subject comprising administering to the subject said compound, so as to thereby inhibit the GSK-3β in the subject.

Description

[18F] MALEIMIDE-BASED GLYCOGEN SYNTHASE KINASE-3BETA LIGANDS FOR POSITRON EMISSION TOMOGRAPHY IMAGING AND RADIOSYNTHESIS METHOD

This application claims priority of U.S. Provisional Application Nos. 62/571,954, filed October 13, 2017 and 62/445,331, filed January 12, 2017, the contents of each of which are hereby incorporated by reference .

Throughout this application, certain publications are referenced in parentheses. Full citations for these publications may be found immediately preceding the claims. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to describe more fully the state of the art to which this invention relates.

BACKGROUND OF THE INVENTION Glycogen synthase kinase-3 (GSK-3) is a serine-threonine protein kinase family consisting of two highly homologous isoforms (GSK-3 and GSK- 3β)^■. This enzyme is ubiquitously expressed in a variety of tissues and is particularly abundant in the central nervous system (CNS) (Woodgett, J. R. 1990) . GSK-3 is associated with a number of cellular functions and physiological events, such as cell proliferation, stem cell renewal, neuropathological events, and apoptosis (Cohen, P. et al. 2001; Kaidanovich-Beilin, O. et al. 2011; Cohen, P. et al . 2004; Jope, R. S. et al. 2004). Dysregulation of GSK-3 activity is linked to numerous severe pathologies, such as diabetes ( Eldar-Finkelman, H. et al. 1999), neurodegenerative conditions, such as Alzheimer' s disease (AD) (Hooper, C. et al. 2008; Avila, J. et al . 2010) and Parkinson disease (Morales-Garcia, J. et al. 2013), psychiatric disorders, such as bipolar disorder and major depression (Pan, J. Q. et a. 2011; Kaidanovich-Beilin, 0. 2012; Valvezan, A. J. et al. 2012), pain (Martins, D. F. 2011; Maixner, D. et al . 2013), cancer (Luo, J. 2009; Martinez, A. et al. 2002; Eldar-Finkelman, H. 2002) and cardiac hypertrophy (Haq, S. et al. 2000; Dorn, G.W. et al. 2005) . Accordingly, GSK~3 has emerged as an important target for development of new drugs lor the treatment of these diseases. However, in order to study GSK- 3β dysregulation in neurological disorders and to evaluate efficacy and mode of action of new Θεκ-3β therapeutic drugs, noninvasive in vivo imaging methods such as positron emission tomography (PET) are needed .

Positron emission tomography (PET) is a noninvasive in vivo imaging technique that uses radiotracers to characterize, visualize, and quantify physiological processes at the molecular level (miller, P.W. et al. 2008) . Development of successful PET tracers for imaging of GSK-3P has been hindered by the ability of potential tracers to pass the blood-brain barrier. Compounds with insufficient brain penetration are unsuitable for neuroimaging purposes. Although a few tracers that can pass the blood-brain-barrier have been reported (Li, L. et al . 2015; Liang, S.H. et al. 2016), they make use of short-lived carbon- 11 (ti/2 = 20.4 min) , which limits their applications to PET imaging centers equipped with an in-house cyclotron.

SUMMARY OF THE INVENTION

The present invention provides a compound having the structure:

wherein

Ri is -H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl- NH₂, alkyl-0 (alkyl) , alkyl-0 (aryl) , alkyl-0 (heteroaryl) , alkyl- NH (alkyl), alkyl- { alkyl ) ₂, alkyl-NH (aryl ) , alkyl-N (aryl) _2/ alkyl- N(aryl) (heteroaryl), alkyl-NH (heteroaryl ) , alkyl- (heteroaryl ) z, alkyl-S (alkyl) , alkyl-S (aryl) , alkyl-S (heteroaryl) , alkylhalide, aikylaryl, alkylheteroaryl or alkyl- (heterocycloalkyl ) ;

R₂, R3, R4, and R are each, independently, -H, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl-NH2, alkyl-0 (alkyl ) , alkyl- NH (alkyl), alkyl- (alkyl) 2, alkylhalide, aikylaryl, alkylheteroaryl, alkyl- (heterocycloalkyl ) , polyhaloalkyl , OH, 0 (alkyl), O(aryl), O (heteroaryl) , O-alkylhalide , O-polyhaloalkyl, NH₂, NH (alkyl), NH(aryl), NH (heteroaryl) , N(alkyl)₂, N (alkyl ) ( aryl) ,

N (alkyl) (heteroaryl ) , N(aryl)₂, (aryl ) (heteroaryl ) , (heteroaryl) 2, SH, S (alkyl), S(aryl), S (heteroaryl ) , O-polyhaloalkyl, CF₃ or SF₅; and A is an unsubstituted or substituted aryl or heteroaryl, wherein the compound contains at least one alkylhalide group, or a salt of the compound. BRIEF DESCRIPTION OF THE FIGURES

Fig . 1 : Spiked QC of [¹⁸F]10a. HPLC column: Eclipse Plus C-18 5 urn, 4.6 X 100 mm, Mobile phase: 50% Acetonitrile/Water, Flow rate: 1.5 ml/min, UV: 254 nm.

Fig . 2 : Time-course of [¹⁸F] -10a activity of the brain and tongue of the rat withand without pretreatment of non-radioactive tracer 10a.

Fig . 3 : Brain PET-CT scans of rat with [¹⁸F]-10a. Images were converted to units of Bq/mL and a scale running from 150k-35k Bq/itiL was used. Rat 2 was pretreated with non-radioactive tracer 10a.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a compound having the structure:

wherein

Ri is -H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl- NH₂, alkyl-0 (alkyl) , alkyl-0 (aryl) , alkyl-0 (heteroaryl ) , alkyl- NH (alkyl), alkyl- (alkyl ) ₂, alkyl-NH (aryl) , alkyl-N (aryl) ₂, alkyl- N(aryl) (heteroaryl), alkyl-NH (heteroaryl ) , alkyl-N (heteroaryl ) i, alkyl-S (alkyl) , alkyl-S ( aryl ) , alkyl-S (heteroaryl ) , alkylhalide, alkylaryl, alkylheteroaryl or alkyl- (heterocycloalkyl) ;

R-2, R3, R¾, and R₅ are each, independently, -H, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl-NH₂, alkyl-0 ( alkyl ) , alkyl- NH (alkyl), alkyl- (alkyl) 2, alkylhalide, alkylaryl, alkylheteroaryl, alkyl- (heterocycloalkyl ) , polyhaloalkyl, OH, 0 (alkyl), O(aryl), 0 (heteroaryl) , O-alkylhalide, O-polyhaloalkyl, NH? , NH (alkyl), NH(aryl), NH (heteroaryl ) , N (alkyl )₂, N ( alkyl ) ( aryl ) ,

N (alkyl ) (heteroaryl ) , N(aryl)₂, (aryl) (heteroaryl) , (heteroaryl) 2, SH, S (alkyl), S(aryl), S (heteroaryl ) , O-polyhaloalkyl, CF₃ or SF$; and A is an unsubstituted or substituted aryl or heteroaryl, wherein the compound contains at least one alkylhalide group, or a salt of the compound.

In some embodiments, the compound wherein

R-. is -H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl- NH:.., alkyl-0 (alkyl) , alkyl- (alkyl) 2, alkylhalide, alkylaryl, alkylheteroaryl or alkyl- (heterocycloalkyl ) ; R:_;, R_J, R-₄, and R₅ are each, independently, H or halogen; and

A is a unsubstituted or substituted aryl or heteroaryl, wherein the compound contains at least one alkylhalide group, or a salt of the compound.

In some embodiments, the compound wherein the compound contains at least one fluoroalkyl group.

In some embodiments, the compound wherein the compound contains at least one -(CH₂)_nF group, wherein n is 2-10.

In some embodiments, the compound wherein A is an unsubstituted or substituted monoaryl or monoheteroaryl.

In some embodiments, the compound wherein A is an unsubstituted or substituted phenyl, thiophene, furan, pyrrole, indole, benzofuran, benzothiophene, oxazole, isoxazole, imidazole, pyrazole, thiazole, isothiazole, triazole, pyrimidine, pyridazine, pyrazine, or pyridine.

In some embodiments, the compound wherein A has the structure:

wherein each of R7, Re, Rs and Rio is, independently, -H, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl-NH₂, alkyl- O(alkyl), alkyl-NH (alkyl) , alkyl- (alkyl ) ₂, alkylhalide, alkylaryl, alkylheteroaryl , alkyl- (heterocycloalkyl) , polyhaloalkyl , OH, 0 (alkyl), O(aryl), 0 ( heteroaryl ) , O-alkylhalide, O-polyhaloalkyl, NH₂, NH(alkyl), NH(aryl), NH (heteroaryl ) , N(alkyl):, ( alkyl ) ( aryl ) , N (alkyl) (heteroaryl) , N(aryl)₂, N ( aryl ) (heteroaryl ) , N (heteroaryl ) 2, SH, S (alkyl), S(aryl), S (heteroaryl ) , O-polyhaloalkyl, or SF5. 13446

In some embodiments, the compound wherein each of R₆, R7, Re, R9 and Rio is, independently, -H, halogen, -OH, -NH2, -CF₃, -O(alkyl), - O(haloalkyl) or -NH(alkyl).

In some embodiments, the compound wherein each of R¾, R₇, R«, R and Rio is, independently, -H, -CI, -OCH3 or -OCH2CH2 F .

In some embodiments, the compound wherein A has the structure:

In some embodiments, the compound wherein one of R6, R₇, R₈, R9 and Rio is -OCH2CH2 F . In some embodiments, the compound wherein A has the structure:

in some embodiments, the compound wherein A is an unsubstituted or substituted thiophene or pyridine.

In some embodiments, the compound wherein A has the structure:

wherein each of Ru, R12, R13, R14 and R15 is, independently, -H, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl-NH₂, alkyl- O(alkyl), alkyl-NH (alkyl ) , alkyl-N (alkyl ) 2, alkylhalide, alkylaryl, alkylheteroaryl, alkyl- (heterocycloalkyl) , polyhaloalkyl , OH, O(alkyl), O(aryl), 0 (heteroaryl ) , O-alkylhalide, O-polyhaloalkyl, NH₂, NH(alkyl), NH(aryl), NH (heteroaryl ) , N(alkyl)₂, (alkyl) (aryl) , N (alkyl) (heteroaryl) , N(aryl)₂, (aryl) (heteroaryl ) , N (heteroaryl ) ₂, SH, S (alkyl), S(aryl), S (heteroaryl ) , O-polyhaloalkyl or SF₅.

In some embodiments, the compound wherein A has the structure:

wherein each of Rie, R17, Ris and R19 is, independently, -H, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl-NH₂, alkyl- 0 (alkyl), alkyl-NH (alkyl ) , alkyl- (alkyl) 2, alkylhalide, alkylaryl, alkylheteroaryl, alkyl- (heterocycloalkyl) , polyhaloalkyl, OH, O(alkyl), O(aryl), 0 (heteroaryl ) , O-alkylhalide, O-polyhaloalkyl, NH₂, NH(alkyl), NH(aryl), NH (heteroaryl ) , N(alkyl)₂, ( alkyl ) ( aryl ) , N (alkyl) (heteroaryl) , N(aryl)₂, ( aryl ) ( heteroaryl ) , N (heteroaryl ) 2, SH, S (alkyl), S(aryl), S (heteroaryl ) , O-polyhaloalkyl or SF₅.

In some embodiments, the compound wherein each of Ru, R:₂, R13, R-.i and Ri5 is, independently, -H, halogen, -OH, -NH? , -CF₃, -O (alkyl), - 0 (alkylhalide) or -NH (alkyl) . In some embodiments, the compound wherein each of Ri , R17, Rie and R19 is, independently, -H, halogen, -OH, - H₂, -CF3, -O(alkyl), 0 (alkylhalide) or -NH(alkyl) . In some embodiments, the compound wherein each of Rn , Ri?, R1.1, R14 and Rib is, independently, -H, -CI, -OCH3 or -OCH2CH2F.

In some embodiments, the compound wherein each of Rie, Rn, Rie and R19 is, independently, -H, -CI, -OCH3 or -OCH₂CH₂F.

In some embodiments, the compound wherein A has the structure:

In some embodiments, the compound wherein Ri is alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl-N¾, alkyl-0 (alkyl) , alkyl , -NH (alkyl ) , alkyl-N (alkyl) 2, alkylhalide, alkylaryl, alkylheteroaryl or alkyl- (heterocycloalkyl) .

In some embodiments, the compound wherein ¾ is alkyl, alkyl-0 ( alkyl ) , alkyl- (alkyl) 2, alkylhalide or alkyl- (heterocycloalkyl) .

In some embodiments, the compound wherein Ri is unbranched C1-C₄ alkyl, C1-C4 alkyl-0 (C1-C4 alkyl), C1-C4 alkyl-N (C1-C4 alkyl) ₂, halo C1-C4 alkyl or Ci-C₄ alkyl- (heterocycloalkyl ) .

In some embodiments, the compound wherein Ri is -CH₃, -CH2CH3, CH₂CH₂CH₃, -CH₂CH₂CH₂CH₃, -CH₂CH?OH, -CH2CH2OCH3, -CH₂CH₂N (CH₃ ) 2 or

In some embodiments, the compound wherein Ri is -CH2CH2F.

In some embodiments, the compound wherein R2, R3, R-i, and R5 are each, independently, is H, F, CI or Br.

In some embodiments, the compound wherein 2, R3, R and R₅ are each, independently, -H, halogen or alkyl.

In some embodiments, the compound wherein R2, R3, 4, and R₅ are each, independently, -H, halogen, alkyl, alkenyl, alkynyl, aryl or heteroaryl .

In some embodiments, the compound wherein R2, R3, R4, and R5 are each, independently, -H, OH, SH, N¾, halogen, alkyl, alkenyl, alkynyl, aryl or heteroaryl.

In some embodiments, the compound wherein the F of the fluoroalkyl group is ¹⁸F.

In some embodiments, the compound wherein Ri is fluoroalkyl or the aryl or heteroaryl Λ is substituted with a fluoroalkyl.

In some embodiments, the compound wherein Ri is -CH2CH₂F or the aryl or heteroaryl A is substituted with a -CH2CH2F.

In some embodiments, the compound wherein the F of the -CH2CH2F group is ¹⁸F.

In some embodiments, the compound having the structure:

a salt of the compound.

some embodiments, the compound having the structure

n some embodiments, the compound having the structure:

or a salt of the compound.

In some embodiments, the compound having the structure:

or a salt of the compound.

In some embodiments, the compound wherein the F of the -CH₂CH2F group is ¹⁸F.

In some embodiments, the compound wherein

Ri is -H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl- NH;, alkyl-0 (alkyl) , alkyl-0 ( aryl ) , alkyl-0 (heteroaryl) , alkyl- NH (alkyl) , alkyl- (alkyl ) ₂, alkyl-NH (aryl) , alkyl-N (aryl) ₂, alkyl- N (aryl ) (heteroaryl ) , alkyl-NH (heteroaryl ) , alkyl-N (heteroaryl ) -_, alkyl-S (alkyl) , alkyl-S { aryl ) , alkyl-S (heteroaryl ) , alkylhalide, alkylaryl, alkylheteroaryl or alkyl- (heterocycloalkyl ) ;

R:-, R;_:, R4, and R5 are each, independently, -H, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl-NH₂, alkyl-0 ( alkyl ) , alkyl- NH (alkyl) , alkyl- (alkyl ) , alkylhalide, alkylaryl, alkylheteroaryl, alkyl- (heterocycloalkyl ) , polyhaloalkyl, OH, O(alkyl), O(aryl), O (heteroaryl) , O-alkylhalide, O-polyhaloalkyl , NH₂, NH (alkyl), NH(aryl), NH (heteroaryl ) , N(alkyl)₂, (alkyl) (aryl) ,

N (alkyl ) (heteroaryl ) , N(aryl)2, N (aryl) (heteroaryl ) , N (heteroaryl ) ₂, SH, 3 (alkyl), S(aryl), S (heteroaryl ) , O-polyhaloalkyl, CF₃ or SF¾; and A is an unsubstituted or substituted aryl or heteroaryl, wherein the compound contains at least one alkylhalide group as Rl or as the substitution on the A group,

or a salt of the compound.

In some embodiments, the compound wherein

Ri is -H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl- NH₂, alkyl-O(alkyl) , alkyl-0 (aryl ) , alkyl-0 (heteroaryl ) , alkyl- NH(alkyl), alkyl-N (alkyl) ₂, alkyl-NH (aryl) , alkyl- ( aryl ) ₂, alkyl- N (aryl ) (heteroaryl ) , alkyl-NH (heteroaryl) , alkyl- (heteroaryl ) ₂, alkyl-S (alkyl) , alkyl-S ( aryl ) , alkyl-S (heteroaryl ) , alkylhalide, alkylaryl, alkylheteroaryl or alkyl- (heterocycloalkyl ) ;

2, R3, R4, and R5 are each, independently, -H or halogen; and

A is an unsubstituted or substituted aryl or heteroaryl,

wherein the compound contains at least one alkylhalide group, or a salt of the compound.

In some embodiments, the compound wherein

Ri is an alkylhalide;

R₂, R3, R4, and R5 are each, independently, -H or halogen; and

A is an unsubstituted or substituted aryl or heteroaryl,

wherein the compound contains at least one alkylhalide group, or a salt of the compound. in some embodiments, the compound wherein

Ri is -H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl- NH-, alkyl-0 (alkyl) , alkyl-0 ( aryl ) , alkyl-0 (heteroaryl ) , alkyl- NH (alkyl), alkyl-N (alkyl) ₂, alkyl-NH (aryl) , alkyl- ( aryl ): , alkyi- N (aryl) (heteroaryl) , alkyl-NH (heteroaryl) , alkyl-N (heteroaryl) 18 013446

16 ^'

alkyl-S (alkyl ) , alkyl-S ( aryl ) , alkyl-S (heteroaryl ) , alkylhalide, alkylaryl, alkylheteroaryl or alkyl- (heterocycloalkyl ) ;

R2, R3, R4, and R5 are each, independently, -H or halogen; and

A is an alkylhalide substituted aryl or heteroaryl,

wherein the compound contains at lcaot ono alkylhalide group, or a salt of the compound.

In some embodiments, the compound wherein

Ri is -H, alkyl, alkenyl, alkyl-OH, alkyl-NHa or alkylhalide;

R2, R3, Ri, and R5 are each, independently, -H or halogen; and

A is an substituted aryl or heteroaryl,

In some embodiments, the compound wherein

Ri is -H, alkyl, alkenyl, alkyl-OH, alkyl-NH₂ or alkylhalide.

In some embodiments, the compound wherein

R?_r Rj, R4, and R are each, independently, -H or halogen. In some embodiments, the compound wherein R₄ is halogen.

In some embodiments, a composition comprising the compound of the present invention or a salt of the compound, and at least one acceptable carrier.

In some embodiments, a pharmaceutical composition comprising the compound of the present invention or a salt of the compound, and at least one acceptable carrier.

In some embodiments, a composition comprising two compounds of the present invention or a salt of the compounds, and at least one acceptable carrier, wherein one compound contains a -CH;CH;F group wherein the F of the -CH₂CH₂F group is ^I8F. 18 013446

17

The present invention provides a method of inhibiting Glycogen synthase kinase-3 p (GSK- 3p ) comprising contacting the Glycogen synthase kinase- 3 with the compound of the present invention, so as to thereby inhibit the GSK-3p .

The present invention provides a method of inhibiting Glycogen synthase kinase- 3 β (GSK-3P) in a subject comprising administering to the subject the compound of the present invention, so as to thereby inhibit the GSK-3 in the subject.

In some embodiments, the method wherein the Glycogen synthase kinase- 3 (GSK-3 ) is located in the brain of the subject.

The present invention provides a method of detecting the presence of Glycogen synthase kinase- 3 (GSK-3p ) in a subject which comprises determining if an amount of the compound of the present invention is present in the subject at a period of time after administration of the compound or salt thereof to the subject, thereby detecting the presence of the Glycogen synthase kinase-3p (GSK- 3p ) based on the amount of the compound determined to be present in the subject.

The present invention provides a method of detecting the presence of Glycogen synthase kinase- 3 (GSR-3 p ) in the brain of a subject which comprises determining if an amount of the compound of the present invention is present in the brain of the subject at a period of time after administration of the compound or salt thereof to the subject, thereby detecting the presence of the Glycogen synthase kinase-3p (GSK- 3 ) based on the amount of the compound determined to be present in the brain of the subject.

The present invention provides a method of detecting the location of Glycogen synthase kinase- 3p (GSK-3P ) in the brain of a subject which comprises determining where an amount of the compound of the present invention is present in the subject at a period of time after administration of the compound or salt thereof to the subject, thereby detecting the location of the Glycogen synthase kmase-SP (GSK-3p) based on the location of the compound determined to be present in the sub ect .

In some embodiments, the method further comprising quantifying the amount of the compound in the subject and comparing the quantity to a predetermined control.

In some embodiments, the method wherein the determining is performed by a Positron Emission Tomography (PET) device.

In some embodiments, the method further comprising determining whether the subject is afflicted with a disease associated with dysregulation of Glycogen synthase kinase-3P (GSK-3p) based on the amount of the compound in the subject.

In some embodiments, the method further comprising determining whether the subject is afflicted with a disease associated with up-regulation of Glycogen synthase kinase-3p (GSK~3 ) based on the amount of the compound in the subject.

In some embodiments, the method further comprising determining whether the subject is afflicted with a disease associated with down- regulation of Glycogen synthase kinase-3p (GSK-3P) based on the amount of the compound in the subject.

In some embodiments, the method wherein the disease associated with dysregulation of Glycogen synthase kinase-3p (GSK-3P) is a neurological disease.

In some embodiments, the method wherein the disease associated with dysregulation of Glycogen synthase kinase-3p (GSK-3P) is Parkinson's disease, Alzheimer's disease (AD), Huntington's disease (HD) , amyotrophic lateral sclerosis (ALS) , bipolar disorder, schizophrenia or major depression.

In some embodiments, the compound contains at least one fluoroalkyl grou . In some embodiments, the compound contains at least one (18)- fluoroalkyl group.

In some embodiments, the compound contains at least one -(CH₂)_nF group, wherein n is 2-10.

In some embodiments, the compound contains at least one -(CH₂)_n ¹⁸F group, wherein n is 2-10.

In some embodiments, the compound contains at least one -(CH2)₂ ¹⁸F group.

In some embodiments, the compound contains one ( 18 ) -fluoroalkyl group.

In some embodiments, the compound contains one -(CH₂)_nF group, wherein n is 2-10.

In some embodiments, the compound contains one -(CHi)_n ^ieF group, wherein n is 2-10. In some embodiments, the compound contains one - (CH-) ;^f,F group.

In some embodiments, the compound wherein A is other than thiophene.

In some embodiments, the compound wherein A is other than 3- hydroxyphenyl or 3-methoxyphenyl .

In some embodiments, the compound wherein A is thiophene. In some embodiments, the compound wherein A is 3-hydroxyphenyl or 3- methoxyphenyl .

In some embodiments of any of the disclosed methods, the compound accumulates in brain colls of the subject.

In some embodiments of any of the disclosed methods, the compound accumulates in brain cells of the subject, wherein the subject is afflicted with is Parkinson's disease, Alzheimer's disease (AD), Huntington's disease (HD) , amyotrophic lateral sclerosis (ALS), bipolar disorder, schizophrenia or major depression.

In some embodiments, a method for the detection of GSK-3 in a subject comprising :

(i) administering to the subject an effective amount of the composition of the present invention;

(ii) allowing a sufficient period of time for brain cells in the subject to take up the compound in the composition; and

(iii) determining whether the GSK-3P is present in the subject by detecting the compound in the subject.

In some embodiments, a method of imaging GSK-3 β in a subject which comprises:

(ii) imaging at least a portion of the subject;

(iii) detecting in the subject the location of the compound, thereby determining the location of the GSK-3 present in the subject based on the location of the compound in the subject;

(v) obtaining an image of the location of the GSK-3P; and optionally the following step: and, optionally,

(vi) repeating steps (i) - (v) one or more times.

In some embodiments, a method of determining the location of GSK-3p in a subject which comprises: (i) administering to the subject an effective amount of the composition of the present invention;

(ii) allowing a sufficient period of time for brain cells in the subject to take up the compound in the composition;

(iii) imaging at least a portion of the subject; and

(iv) detecting in the subject the location of the compound, thereby determining the location of the GSK-3P present in the subject based on the location of the compound in the subject.

In one embodiment, a process for manufacturing a composition which comprises obtaining the compound of the present invention and combining the compound with a carrier so as to thereby manufacture the composition.

In one embodiment, a process for manufacturing a composition which comprises obtaining the ¹⁸F labeled compound of the present invention and combining the compound with a carrier so as to thereby manufacture the composition.

As used herein, a "symptom" associated with a disease or disorder includes any clinical or laboratory manifestation associated with the disease or disorder and is not limited to what the subject can feel or observe.

As used herein, "treating", e.g. of an infection, encompasses inducing prevention, inhibition, regression, or stasis of the disease or a symptom or condition associated with the infection.

The compounds of the present invention include all hydrates, solvates, and complexes of the compounds used by this invention. If a chiral center or another form of an isomeric center is present in a compound of the present invention, all forms of such isomer or isomers, including enantiomers and diastereomers , are intended to be covered herein. Compounds containing a chiral center may be used as a racemic mixture, an enantiomerically enriched mixture, or the racemic mixture may be separated using well-known techniques and an individual enantiomer may be used alone. The compounds described in the present invention are in racemic form or as individual enantiomers. The enantiomers can be separated using known techniques, such as those described in Pure and Applied Chemistry 69, 1469-1474, (1997) IUPAC. In cases in which compounds have unsaturated carbon-carbon double bonds, both the cis-(Z) and trans- (E) isomers are within the scope of this invention.

The compounds of the subject invention may have spontaneous tautomeric forms. In cases wherein compounds may exist in tautomeric forms, such as keto-enol tautomers, each tautomeric form is contemplated as being included within this invention whether existing in equilibrium or predominantly in one form.

In the compound structures depicted herein, hydrogen atoms are not shown for carbon atoms having less than four bonds to non-hydrogen atoms. However, it is understood that enough hydrogen atoms exist on said carbon atoms to satisfy the octet rule.

This invention also provides isotopic variants of the compounds disclosed herein, including wherein the isotopic atom is ²H and/or wherein the isotopic atom ¹³C and/or wherein the isotopic atom ¹⁸F. Accordingly, in the compounds provided herein hydrogen can be enriched in the deuterium isotope. It is to be understood that the invention encompasses all such isotopic forms.

It is understood that the structures described in the embodiments of the methods hereinabove can be the same as the structures of the compounds described hereinabove.

It is understood that where a numerical range is recited herein, the present invention contemplates each integer between, and including, the upper and lower limits, unless otherwise stated.

Except where otherwise specified, if the structure of a compound of this invention includes an asymmetric carbon atom, it is understood that the compound occurs as a racemate, racemic mixture, and isolated single enantiomer. All such isomeric forms of these compounds are 13446

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expressly included in this invention. Except where otherwise specified, each stereogenic carbon may be of the R or S configuration. It is to be understood accordingly that the isomers arising from such asymmetry (e.g., all enantiomers and diastereomers ) are included within the scope of this invention, unless indicated otherwise. Such isomers can be obtained in substantially pure form by classical separation techniques and by stereochemically controlled synthesis, such as those described in "Enantiomers, Racemates and Resolutions" by J. Jacques, A. Collet and S. Wilen, Pub. John Wiley & Sons, NY, 1981 . For example, the resolution may be carried out by preparative chromatography on a chiral column.

The subject invention is also intended to include all isotopes of atoms occurring on the compounds disclosed herein. Isotopes include those atoms having the same atomic number but different mass numbers. By way of general example and without limitation, isotopes of hydrogen include tritium and deuterium. Isotopes of carbon include C-13 and C- 14 . It will be noted that ^'any notation of a carbon in structures throughout this application, when used without further notation, are intended to represent all isotopes of carbon, such as ¹²C, ¹³C, or ¹⁴C. Furthermore, any compounds containing ¹³C or ¹⁴C may specifically have the structure of any of the compounds disclosed herein.

It will also be noted that any notation of a hydrogen in structures throughout this application, when used without further notation, are intended to represent all isotopes of hydrogen, such as ^:H, ²H, or ³H. Furthermore, any compounds containing ²H or ³H may specifically have the structure of any of the compounds disclosed herein.

It will also be noted that any notation of a flourines (F) in structures throughout this application, when used without further notation, are intended to represent all isotopes of fluorine, such as I 9F or ¹⁸F. Furthermore, any compounds containing ¹⁹F or ¹⁸F may 6

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specifically have the structure of any of the compounds disclosed herein .

Isotopically-labeled compounds can generally be prepared by conventional techniques known to those skilled in the art using appropriate isotopically-labeled reagents in place of the non-labeled reagents employed.

In the compounds used in the method of the present invention, the substituents may be substituted or unsubstituted, unless specifically defined otherwise.

It is understood that substituents and substitution patterns on the compounds used in the method of the present invention can be selected by one of ordinary skill in the art to provide compounds that are chemically stable and that can be readily synthesized by techniques known in the art from readily available starting materials. If a substituent is itself substituted with more than one group, it is understood that these multiple groups may be on the same carbon or on different carbons, so long as a stable structure results.

In choosing the compounds used in the method of the present invention, one of ordinary skill in the art will recognize that the various substituents, i.e. R-_I; R2, etc. are to be chosen in conformity with well-known principles of chemical structure connectivity.

As used herein, "alkyl" includes both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms and may be unsubstituted or substituted. Thus, Ci-C_n as in "Ci-C_n alkyl" is defined to include individual groups each having 1, 2...., n-1 or n carbons in a linear or branched arrangement. For example, Ci-Cc, as in "C--Ce. alkyl" is defined to include individual groups each having 1, 2, 3, 4, 5, or 6 carbons in a linear or branched arrangement, and specifically includes methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, pentyl, hexyl, and octyl . As used herein, "alkenyl" refers to a non-aromatic hydrocarbon radical, straight or branched, containing at least 1 carbon to carbon double bond, and up to the maximum possible number of non-aromatic carbon-carbon double bonds may be present, and may be unsubstituted or substituted. For example, "Cz-Cc alkenyl" means an alkenyl radical having 2, 3, 4, 5, or 6 carbon atoms, and up to 1, 2, 3, 4, or 5 carbon-carbon double bonds respectively. Alkenyl groups include ethenyl, propenyl, butenyl and cyclohexenyl . The term "alkynyl" refers to a hydrocarbon radical, straight or branched, containing at least 1 carbon to carbon triple bond, and up to the maximum possible number of non-aromatic carbon-carbon triple bonds may be present, and may be unsubstituted or substituted. Thus, "C₂-C6 alkynyl" means an alkynyl radical having 2 or 3 carbon atoms and 1 carbon-carbon triple bond, or having 4 or 5 carbon atoms and up to 2 carbon-carbon triple bonds, or having 6 carbon atoms and up to 3 carbon-carbon triple bonds. Alkynyl groups include ethynyl, propynyl and butynyl. As herein, "cycloalkyl" shall mean cyclic rings of alkanes of three to eight total carbon atoms, or any number within this range (i.e., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl ) . As used herein, "aryl" is intended to mean any stable monocyclic, bicyclic or polycyclic carbon ring of up to 10 atoms in each ring, wherein at least one ring is aromatic, and may be unsubstituted or substituted. Examples of such aryl elements include phenyl, p-toluenyl ( -methylphenyl ) , naphthyl, tetrahydro-naphthyl, indanyl, biphenyl, phenanthryl, anthryl or acenaphthyl . In cases where the aryl substituent is bicyclic and one ring is non-aromatic, it is understood that attachment is via the aromatic ring.

The term "alkylaryl" refers to alkyl groups as described above wherein one or more bonds to hydrogen contained therein are replaced by a bond to an aryl group as described above. It is understood that an 2018/013446

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"arylalkyl" group is connected to a core molecule through a bond from the alkyl group and that the aryl group acts as a substituent on the alkyl group. Examples of arylalkyl moieties include, but are not limited to, benzyl (phenylmethyl ) , p-trifluoromethylbenzyl (4- tririuoromethyl-phenylmcthyl ) , 1-phenylethyl, 2-phenyl ethyl , 3- phenylpropyl, 2-phenylpropyl and the like.

The term "heteroaryl", as used herein, represents a stable monocyclic, bicyclic or polycyclic ring of up to 10 atoms in each ring, wherein at least one ring is aromatic and contains from 1 to 4 heteroatoms selected from the group consisting of O, N and S. Bicyclic aromatic heteroaryl groups include phenyl, pyridine, pyrimidine or pyridizine rings that are (a) fused to a 6-membered aromatic (unsaturated) heterocyclic ring having one nitrogen atom; (b) fused to a 5- or 6- membered aromatic (unsaturated) heterocyclic ring having two nitrogen atoms; (c) fused to a 5-membered aromatic (unsaturated) heterocyclic ring having one nitrogen atom together with either one oxygen or one sulfur atom; or (d) fused to a 5-membered aromatic (unsaturated) heterocyclic ring having one heteroatom selected from 0, N or S. Heteroaryl groups within the scope of this definition include but are not limited to: benzoimidazolyl , benzofuranyl, benzofurazanyl, benzopyrazolyl , benzotriazolyl , benzothiophenyl , benzoxazolyl, carbazolyl, carbolinyl, cinnolinyl, furanyl, indolinyl, indolyl, indolazinyl, indazolyl, isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl , isoxazolyl, naphthpyridinyl , oxadiazolyl, oxazolyl, oxazoline, isoxazoline, oxetanyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridopyridinyl , pyridazinyl, pyridyl, pyrimidyl, pyrrolyl, quina zolinyl , quinolyl, quinoxalinyl , tetrazolyl, tetrazolopyridyl , thiadiazolyl , thiazolyl, thienyl, triazolyl, azetidinyl, aziridmyl, 1, 4-dioxanyl, hexahydroazepinyl, dihydrobenzoimidazolyl , dihydrobenzofuranyl , dihydrobenzothiophenyl , dihydrobenzoxazolyl, dihydrofuranyl , dihydroimidazolyl, dihydroindolyl , dihydroisooxazolyl , dihydroisothiazolyl, dihydrooxadiazolyl , dihydrooxazolyl , dihydropyrazinyl , dihydropyrazolyl, dihydropyridinyl , dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl , dihydrotetrazolyl , 6

27 dihydrothiadiazolyl, dihydrothiazolyl , dihydrothienyl, dihydrotriazolyl , dihydroazetidinyl , methylenedioxybenzoyl, tetrahydrofuranyl, tetrahydrothienyl, acridinyl, carbazolyl, cinnolinyl, quinoxalinyl , pyrrazolyl, indolyl, benzotriazolyl, benzothiazolyl , benzoxazolyl, isox zolyl, isothiazolyl, furanyl, thienyl, benzothionyl , benzofuranyl , qui nol i yl , isoquinolinyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, tetra-hydroquinoline . In cases where the heteroaryl substituent is bicyclic and one ring is non-aromatic or contains no heteroatoms, it is understood that attachment is via the aromatic ring or via the heteroatom containing ring, respectively. If the heteroaryl contains nitrogen atoms, it is understood that the corresponding N-oxides thereof are also encompassed by this definition .

The term "alkylheteroaryl" refers to alkyl groups as described above wherein one or more bonds to hydrogen contained therein are replaced by a bond to an heteroaryl group as described above. It is understood that an "alkylheteroaryl" group is connected to a core molecule through a bond from the alkyl group and that the heteroaryl group acts as a substituent on the alkyl group. Examples of alkylheteroaryl moieties include, but are not limited to, -CH₂- ( C₅H₄N) , -CH₂-CH₂- (C₅H₄N) and the like. The term "heterocycle" or "heterocyclylic" refers to a mono- or poly- cyclic ring system which can be saturated or contains one or more degrees of unsaturation and contains one or more heteroatoms. Preferred heteroatoms include N, 0, and/or S, including N-oxides, sulfur oxides, and dioxides. Preferably the ring is three to ten- membered and is either saturated or has one or more degrees of unsaturation. The heterocycle may be unsubstituted or substituted, with multiple degrees of substitution being allowed. Such rings may be optionally fused to one or more of another "heterocyclic" ring(s), heteroaryl ring(s), aryl ring(s), or cycloalkyl ring(s). Examples of heterocycles include, but are not limited to, tetrahydrofuran, pyran, 1,4-dioxane, 1,3-dioxane, piperidine, piperazine, pyrrolidine, 2018/013446

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morpholine, thiomorpholine, tetrahydrothiopyran, tetrahydrothiophene, 1 , 3-oxathiolane, imidazole, tetrahydropyran, dihydropiperidine, tetrahydrothiophene and the like. The alkyl, alkcnyl, alkynyl, aryl, heteroaryl and heterocyclyl subsLituents may be substituted or unsubstituted, unless specif cally defined otherwise. In the compounds of the present invention, alkyl, alkenyl, alkynyl, aryl, heterocyclyl and heteroaryl groups can be further substituted by replacing one or more hydrogen atoms with alternative non-hydrogen groups. These include, but are not limited to, halo, hydroxy, mercapto, amino, carboxy, cyano and carbamoyl.

As used herein, the term "halogen" refers to F, Cl, Br, and I. The term "alkylhalide" refers to alkyl groups as described above wherein one or more bonds to hydrogen contained therein are replaced by a bond to a halogen.

The terms "substitution", "substituted" and "substituent" refer to a functional group as described above in which one or more bonds to a hydrogen atom contained therein are replaced by a bond to non-hydrogen or non-carbon atoms, provided that normal valencies are maintained and that the substitution results in a stable compound. Substituted groups also include groups in which one or more bonds to a carbo (s) or hydrogen (s) atom are replaced by one or more bonds, including double or triple bonds, to a heteroatom. Examples of substituent groups include the functional groups described above, and halogens (i.e., F, Cl, Br, and I); alkyl groups, such as methyl, ethyl, n- propyl, isopropryl, n-butyl, tert-butyl, and trifluoromethyl; hydroxyl; alkoxy groups, such as methoxy, ethoxy, n-propoxy, and isopropoxy; aryloxy groups, such as phenoxy arylalkyloxy, such as benzyloxy (phenylmethoxy ) and p-trifluoromethylbenzyloxy (4- trifluoromethylphenylmethoxy ) ; heteroaryloxy groups; sulfonyl groups, such as trifluoromethanesulfonyl, methanesul fonyl , and p- toluenesulfonyl; nitro, nitrosyl; mercapto; sulfanyl groups, such as methylsulfanyl, ethylsulfanyl and propyl sulfanyl ; cyano; amino groups, such as amino, methylamino, dimethylamino, ethylamino, and diethylamino; and carboxyl . Where multiple substituent moieties are disclosed or claimed, the substituted compound can be independently substituted by one or more of the disclosed or claimed substituent moietie3, singly or pluraly. By independently .substituted, it is meant that the (two or more) substituents can be the same or different.

In choosing the compounds used in the method of the present invention, one of ordinary skill in the art will recognize that the various substituents, i.e. Ri, R2, etc. are to be chosen in conformity with well-known principles of chemical structure connectivity.

In the compounds of the present invention, the substituents may be substituted or unsubstituted, unless specifically defined otherwise.

In the compounds of the present invention, alkyl, heteroalkyl, monocycle, bicycle, aryl, heteroaryl and heterocycle groups can be further substituted by replacing one or more hydrogen atoms with alternative non-hydrogen groups. These include, but are not limited to, halo, hydroxy, mercapto, amino, carboxy, cyano, carbamoyl and aminocarbonyl and aminothiocarbonyl .

It is understood that substituents and substitution patterns on the compounds of the instant invention can be selected by one of ordinary skill in the art to provide compounds that are chemically stable and that can be readily synthesized by techniques known in the art, as well as those methods set forth below, from readily available starting materials. If a substituent is itself substituted with more than one group, it is understood that these multiple groups may be on the same carbon or on different carbons, so long as a stable structure results.

In choosing the compounds of the present invention, one of ordinary skill in the art will recognize that the various substituents, i.e. Ri, R_j, etc. are to be chosen in conformity with well-known principles of chemical structure connectivity. The various R groups attached to the aromatic rings of the compounds disclosed herein may be added to the rings by standard procedures, for example those set forth in Advanced Organic Chemistry: Part B: Reaction and Synthesis, Francis Carey and Richard Sundberg, (Springer) 5th cd. Edition. (2007), the content of which i,s hereby incorporated by reference.

The compounds used in the method of the present invention may be prepared by techniques well known in organic synthesis and familiar to a practitioner ordinarily skilled in the art. However, these may not be the only means by which to synthesize or obtain the desired compounds .

The compounds used in the method of the present invention may be prepared by techniques described in Vogel's Textbook of Practical Organic Chemistry, A.I. Vogel, A.R. Tatchell, B.S. Furnis, A.J. Hannaford, P.W.G. Smith, (Prentice Hall) 5^th Edition (1996), March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, Michael B. Smith, Jerry March, (Wiley-Interscience) 5^th Edition (2007), and references therein, which are incorporated by reference herein. However, these may not be the only means by which to synthesize or obtain the desired compounds.

Another aspect of the invention comprises a compound of the present invention as a pharmaceutical composition.

In some embodiments, a pharmaceutical composition comprising the compound of the present invention and a pharmaceutically acceptable carrier .

As used herein, the term "pharmaceutically active agent" means any substance or compound suitable for administration to a subject and furnishes biological activity or other direct effect in the treatment, cure, mitigation, diagnosis, or prevention of disease, or affects the structure or any function of the subject. Pharmaceutically active agents include, but are not limited to, substances and compounds described in the Physicians' Desk Reference (PDR Network, LLC; 64th edition; November 15, 2009) and "Approved Drug Products with Therapeutic Equivalence Evaluations" (U.S. Department Of Health And Human Services, 30^th edition, 2010) , which are hereby incorporated by reference. Pharmaceutically active agents which have pendant carboxylic acid groups may be modified in accordance with the present invention using standard esterification reactions and methods readily available and known to those having ordinary skill in the art of chemical synthesis. Where a pharmaceutically active agent does not possess a carboxylic acid group, the ordinarily skilled artisan will be able to design and incorporate a carboxylic acid group into the pharmaceutically active agent where esterification may subsequently be carried out so long as the modification does not interfere with the pharmaceutically active agent's biological activity or effect.

The compounds used in the method of the present invention may be in a salt form. As used herein, a "salt" is a salt of the instant compounds which has been modified by making acid or base salts of the compounds. In the case of compounds used to treat an infection or disease caused by a pathogen, the salt is pharmaceutically acceptable. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as phenols. The salts can be made using an organic or inorganic acid. Such acid salts are chlorides, bromides, sulfates, nitrates, phosphates, sulfonates, formates, tartrates, maleates, malates, citrates, benzoates, salicylates, ascorbates, and the like. Phenolate salts are the alkali earth metal salts, sodium, potassium or lithium. The term "pharmaceutically acceptable salt" in this respect, refers to the relatively non-toxic, inorganic and organic acid or base addition salts of compounds of the present invention. These salts can be prepared in situ during the final isolation and purification of the compounds of the invention, or by separately reacting a purified compound of the invention in its free base or free acid form with a suitable organic or inorganic acid or base, and isolating the salt thus formed. Representative salts include the hydrobromide , hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napthylate, mesylate, glucoheptonate, lactobionate, and laurylsulphonate salts and the like. (See, e.g., Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19).

As used herein, "administering" an agent may be performed using any of the various methods or delivery systems well known to those skilled in the art. The administering can be performed, for example, orally, parenterally, intraperitoneally, intravenously, intraarterially, transdermally, sublingually, intramuscularly, rectally, transbuccally, intranasally, liposomally, via inhalation, vaginally, intraoccularly, via local delivery, subcutaneously, intraadiposally, intraarticularly, intrathecally, into a cerebral ventricle, intraventicularly, intratumorally, into cerebral parenchyma or intraparenchchymally .

As used herein, a "pharmaceutically acceptable carrier" is a pharmaceutically acceptable solvent, suspending agent or vehicle, for delivering the instant compounds to the animal or human. The carrier may be liquid or solid and is selected with the planned manner of administration in mind. Liposomes are also a pharmaceutically acceptable carrier as are slow-release vehicles .

The dosage of the compounds administered in treatment will vary depending upon factors such as the pharmacodynamic characteristics of a specific chemotherapeutic agent and its mode and route of administration; the age, sex, metabolic rate, absorptive efficiency, health and weight of the recipient; the nature and extent of the symptoms; the kind of concurrent treatment being administered; the frequency of treatment with; and the desired therapeutic effect.

The compounds can be administered in oral dosage forms as tablets, capsules, pills, powders, granules, elixirs, tinctures, suspensions, syrups, and emulsions. The compounds may also be administered in intravenous (bolus or infusion) , intraperitoneal, subcutaneous, or intramuscular form, or introduced directly, e.g. by injection, topical application, or other methods, into or topically onto a site of disease or lesion, all using dosage forms well known to those of ordinary skill in the pharmaceutical arts.

The compounds used in the method of the present invention nan be administered in admixture with suitable pharmaceutical diluents, extenders, excipients, or in carriers such as the novel programmable sustained-release multi-compartmental nanospheres (collectively referred to herein as a pharmaceutically acceptable carrier) suitably selected with respect to the intended form of administration and as consistent with conventional pharmaceutical practices. The unit will be in a form suitable for oral, nasal, rectal, topical, intravenous or direct injection or parenteral administration. The compounds can be administered alone or mixed with a pharmaceutically acceptable carrier. This carrier can be a solid or liquid, and the type of carrier is generally chosen based on the type of administration being used. The active agent can be co-administered in the form of a tablet or capsule, liposome, as an agglomerated powder or in a liquid form. Examples of suitable solid carriers include lactose, sucrose, gelatin and agar. Capsule or tablets can be easily formulated and can be made easy to swallow or chew; other solid forms include granules, and bulk powders. Tablets may contain suitable binders, lubricants, diluents, disintegrating agents, coloring agents, flavoring agents, flow- inducing agents, and melting agents. Examples of suitable liquid dosage forms include solutions or suspensions in water, pharmaceutically acceptable fats and oils, alcohols or other organic solvents, including esters, emulsions, syrups or elixirs, suspensions, solutions and/or suspensions reconstituted from non-effervescent granules and effervescent preparations reconstituted from effervescent granules. Such liquid dosage forms may contain, for example, suitable solvents, preservatives, emulsifying agents, suspending agents, diluents, sweeteners, thickeners, and melting agents. Oral dosage forms optionally contain flavorants and coloring agents. Parenteral and intravenous forms may also include minerals P T/US2018/013446

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and other materials to make them compatible with the type of injection or delivery system chosen.

Techniques and compositions for making dosage forms useful in the present invention are described in the following references: 7 Modern Pharmaceutics, Chapters 9 and 10 (Banker f, Rhodes, Editors, 1979); Pharmaceutical Dosage Forms: Tablets (Lieberman et al . , 1981); Ansel, Introduction to Pharmaceutical Dosage Forms 2nd Edition (1976) ; Remington's Pharmaceutical Sciences, 17th ed. (Mack Publishing Company, Easton, Pa., 1985); Advances in Pharmaceutical Sciences (David Ganderton, Trevor Jones, Eds., 1992); Advances in Pharmaceutical Sciences Vol. 7. (David Ganderton, Trevor Jones, James McGinity, Eds., 1995); Aqueous Polymeric Coatings for Pharmaceutical Dosage Forms (Drugs and the Pharmaceutical Sciences, Series 36 (James McGinity, Ed., 1989); Pharmaceutical Particulate Carriers: Therapeutic Applications: Drugs and the Pharmaceutical Sciences, Vol 61 (Alain Rolland, Ed., 1993); Drug Delivery to the Gastrointestinal Tract (Ellis Horwood Books in the Biological Sciences. Series in Pharmaceutical Technology; J. G. Hardy, S. S. Davis, Clive G. Wilson, Eds.); Modem Pharmaceutics Drugs and the Pharmaceutical Sciences, Vol 40 (Gilbert S. Banker, Christopher T. Rhodes, Eds.) . All of the aforementioned publications are incorporated by reference herein.

Tablets may contain suitable binders, lubricants, disintegrating agents, coloring agents, flavoring agents, flow-inducing agents, and melting agents. For instance, for oral administration in the dosage unit form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, gelatin, agar, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth, or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

The compounds used in the method of the present invention may also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids such as lecithin, sphingomyelin, proteolipids , protein-encapsulated vesicles or from cholesterol, stearylamine, or phosphatidylcholines. The compounds may be administered as components of tissue-targeted emulsions .

The compounds used in the method of the present invention may also be coupled to soluble polymers as targetable drug carriers or as a prodrug. Such polymers include polyvinylpyrrolidone, pyran copolymer, polyhydroxylpropylmethacrylamide-phenol , polyhydroxyethylasparta- midephenol, or polyethyleneoxide-polylysine substituted with palmitoyl residues. Furthermore, the compounds may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters , polyacetals, polydihydropyrans , polycyanoacylates , and crosslinked or amphipathic block copolymers of hydrogels.

Gelatin capsules may contain the active ingredient compounds and powdered carriers, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as immediate release products or as sustained release products to provide for continuous release of medication over a period of hours. Compressed tablets can be sugar-coated or film-coated to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric coated for selective disintegration in the gastrointestinal tract . For oral administration in liquid dosage form, the oral drug components are combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Examples of suitable liquid dosage forms include solutions or suspensions in water, pharmaceutically acceptable fats and oils, alcohols or other organic solvents, including esters, emulsions, syrups or elixirs, suspensions, solutions and/or suspensions reconstituted from non-effervescent granules and effervescent preparations reconstituted from effervescent granules. Such liquid dosage forms may contain, for example, suitable solvents, preservatives, emulsifying agents, suspending agents, diluents, sweeteners, thickeners, and melting agents.

Liquid dosage forms for oral administration can contain coloring and flavoring to increase patient acceptance. In general, water, asuitable oil, saline, aqueous dextrose (glucose) , and related sugar solutions and glycols such as propylene glycol or polyethylene glycols are suitable carriers for parenteral solutions. Solutions for parenteral administration preferably contain a water soluble salt of the active ingredient, suitable stabilizing agents, and if necessary, buffer substances. Antioxidizing agents such as sodium bisulfite, sodium sulfite, or ascorbic acid, either alone or combined, are suitable stabilizing agents. Also used are citric acid and its salts and sodium EDTA. In addition, parenteral solutions can contain preservatives, such as benzalkonium chloride, methyl- or propyl-paraben, and chlorobutanol . Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, Mack Publishing Company, a standard reference text in this field.

The compounds used in the method of the present invention may also be administered in intranasal form via use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art. To be administered in the form of a transdermal delivery system, the dosage administration will generally be continuous rather than intermittent throughout the dosage regimen.

Parenteral and intravenous forms may also include minerals and other materials such as solutol and/or othanol to make them compatible with the type of injection or delivery 3yotcm chosen.

Specific examples of pharmaceutically acceptable carriers and excipients that may be used to formulate oral dosage forms of the present invention are described in U.S. Pat. No. 3,903,297 to Robert, issued Sept. 2, 1975. Techniques and compositions for making dosage forms useful in the present invention are described-in the following references: 7 Modern Pharmaceutics, Chapters 9 and 10 (Banker & Rhodes, Editors, 1979); Pharmaceutical Dosage Forms: Tablets (Lieberman et al . , 1981); Ansel, Introduction to Pharmaceutical Dosage Forms 2nd Edition (1976); Remington's Pharmaceutical Sciences, 17th ed. (Mack Publishing Company, Easton, Pa., 1985); Advances in Pharmaceutical Sciences (David Ganderton, Trevor Jones, Eds., 1992); Advances in Pharmaceutical Sciences Vol 7. (David Ganderton, Trevor Jones, James McGinity, Eds., 1995); Aqueous Polymeric Coatings for Pharmaceutical Dosage Forms (Drugs and the Pharmaceutical Sciences, Series 36 (James McGinity, Ed., 1989); Pharmaceutical Particulate Carriers : Therapeutic Applications : Drugs and the Pharmaceutical Sciences, Vol 61 (Alain Rolland, Ed., 1993); Drug Delivery to the Gastrointestinal Tract (Ellis Horwood Books in the Biological Sciences. Series in Pharmaceutical Technology; J. G. Hardy, S. S. Davis, Clive G. Wilson, Eds.); Modem Pharmaceutics Drugs and the Pharmaceutical Sciences, Vol 40 (Gilbert S. Banker, Christopher T. Rhodes, Eds.). All of the aforementioned publications are incorporated by reference herein.

Solid dosage forms, such as capsules and tablets, may be enteric- coated to prevent release of the active ingredient compounds before they reach the small intestine. Materials that may be used as enteric coatings include, but are not limited to, sugars, fatty acids, proteinaceous substances such as gelatin, waxes, shellac, cellulose acetate phthalate (CAP), methyl acrylate-methacrylic acid copolymers, cellulose acetate succinate, hydroxy propyl methyl cellulose phthalate, hydroxy propyl methyl cellulose acetate succinate (hypromellose acetate succinate), polyvinyl acetate phthalate (PVAP), and methyl methacrylaLe-iuethacrylic acid copolymers .

Variations on those general synthetic methods will be readily apparent to those of ordinary skill in the art and are deemed to be within the scope of the present invention.

Each embodiment disclosed herein is contemplated as being applicable to each of the other disclosed embodiments. Thus, all combinations of the various elements described herein are within the scope of the invention.

This invention will be better understood by reference to the Experimental Details which follow, but those skilled in the art will readily appreciate that the specific experiments detailed are only illustrative of the invention as described more fully in the claims which follow thereafter.

EXPERIMENTAL DETAILS

All air- and moisture-insensitive reactions were carried out under an ambient atmosphere, magnetically stirred, and monitored by thin layer chromatography (TLC) using Agela Technologies TLC plates pre-coated with 250 ym thickness silica gel 60 F254 plates and visualized by fluorescence quenching under UV light. Flash chromatography was performed on SiliaFlash® Silica Gel 40-63 μπι 60 A particle size using a forced flow of eluent at 0.3-0.5 bar pressure (Still, W. et al . 1978) . All air- and moisture-sensitive manipulations were performed using oven-dried glassware, including standard Schlenk and glovebox techniques under an atmosphere of nitrogen. Diethyl ether and THF were distilled from deep purple sodium benzophenone ketyl . Methylene chloride, chloroform and acetonitrile were dried over Ca¾ and distilled. Methylene chloride was degassed via three freeze-pump-thaw cycles. All other chemicals were used as received. All deuterated solvents were purchased from Cambridge Isotope Laboratories.

NMR spectra were recorded on either a Bruker Ascend 700 spectrometer operating at 700 MHz for ¾ acquisitions and 175 MHz for ¹³C acquisitions, a Bruker 500 Advance spectrometer operating at 500 MHz and 125 MHz for ¹H and ¹³C acquisitions, respectively, a Bruker 400 Nanobay spectrometer operating at 400 MHz, 100 MHz, and 376 MHz for ^ΊΗ, ¹³C, and ¹⁹F acquisitions, respectively. Chemical shifts were referenced to the residual proton solvent peaks (¹H: CDCI₃ , δ 7.26; (CD₃)₂SO, δ 2.50; CD₃OD, δ 3.31; CD₃CN, δ 1.94), solvent ¹³C signals ( CDCI₃ , δ 77.16; (CD₃)₂SO, δ 39.52; CD₃OD, δ 49.00) (Fulmer, G. et al . 2010), dissolved or external neat PhCF₃ (¹⁹F, δ -63.3 relative to C FC-. ) (Wang, X. et al. 2013) . Signals are listed in ppm, and multiplicity identified as s = singlet, br = broad, d = doublet, t = triplet, q = quartet, m = multiplet; coupling constants in Hz; integration. High- resolution mass spectra were performed at Mass Spectrometry Services at the Univ. of Illinois at Urbana-Champaign and were obtained using Waters Q-TOF Ultima ESI mass spectrometer. Concentration under reduced pressure was performed by rotary evaporation at 25-30 °C at appropriate pressure. Purified compounds were further dried under high vacuum (0.01-0.05 Torr) . Yields refer to purified and spectroscopically pure compounds. GE Tracerlab FXF_N synthesis module and Waters QMA cartridge were used for the radiosynthesis of [¹⁸F]10a.

Methyl 2- (5-fluoro-lH-indol-3-yl) -2-oxoacetate (5a)

To a solution of 5-fluoro-lff-indole (2.00 g, 14.8 mmol, 1.00 equiv) in Et₂0 (20.0 mL, 0.740 M) was added oxalyl chloride (1.80 mL, 21.1 mmol, 1.43 equiv) dropwise at 0 °C. The yellow slurry was stirred at 0 °C for 30 min and then cooled to -78 °C. A solution of NaOMe in MeOH (25%, 8.00 mL) was added to this slurry at the same temperature. The reaction mixture was then allowed to warm up to room temperature, and was quenched by addition of water (10.0 mL) . The precipitate was collected by filtration, washed with water and dried to afford the title compound as a yellow solid (2.85 g, 12.9 mmol, 87% yield) .

NMR Spectroscopy: *H NMR (700 MHz, (CD₃)₂SO, 25 °C, δ) : 12.54 (s, 1H) , 8.51 (s, 1H) , 7.82 (s, 1H) , 7.56 (s, 1H) , 7.12-7.20 (m, 1H) , 3.89 (s, 3H) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ) : 178.4, 163.6, 159.2 (d, J = 232.8 Hz), 139.7, 133.3, 126.3 (d, J = 10.5 Hz), 114.2 (d, J = 10.5 Hz), 112.5 (d, J = 5.3 Hz), 112.0 (d, J = 26.3 Hz), 106.2 (d, J = 24.5 Hz), 52.6. The ¹H NMR data were in good agreement with values reported in the literature (Ye, Q. et al . 2015) .

Methyl 2- 5-chloro-lH-indol-3-yl) -2-oxoacetate (5b)

To a solution of 5-chloro-lii-indole (2.80 g, 18.5 mmol, 1.00 equiv) in Et₂0 (25.0 mL, 0.740 M) was added oxalyl chloride (1.89 mL, 22.2 mmol, 1.22 equiv) dropwise at 0 °C. The yellow slurry was stirred at 0 °C for 30 min and then cooled to -78 °C. A solution of NaOMe in MeOH (25%, 8.50 mL) was added to this slurry at the same temperature. The reaction mixture was then allowed to warm up to room temperature, and wa3 quenched by addition of water (10.0 mL) . The precipitate was collected by filtration, washed with water and dried to afford the title compound as a gray solid (4.00 g, 16.8 mmol, 91% yield).

NMR Spectroscopy: *H NMR (500 MHz, (CD₃)₂SO, 25 °C, δ) : 12.71 (s, 1H) , 8.52 (d, J = 3.3 Hz, 1H) , 8.13 (d, J = 2.1 Hz, 1H) , 7.58 (d, J = 8.6 Hz, 1H), 7.32 (dd, J = 2.1, 8.6 Hz, 1H) , 3.89 (s, 3H) . ¹³C NMR (125 MHz, (CD₃)₂SO, 25 °C, δ): 178.4, 163.4, 139.5, 135.2, 127.5, 126.8, 123.9, 120.2, 114.5, 112.0, 52.6. The ^ΧΗ NMR data were in good agreement with values reported in the literature (Ye, Q. et al . 2015) . Methyl 2- (lH-indol-3-yl) -2-oxoacetate (5c)

To a solution of lH-indole (6.00 g, 51.2 mmol, 1.00 equiv) in Et₂0 (60.0 mL, 0.850 M) was added oxalyl chloride (5.27 mL, 61.4 mmol, 1.20 equiv) dropwise at 0 °C. The yellow slurry was stirred at 0 °C for 30 min and then cooled to -78 °C. A solution of NaOMe in MeOH (25%, 23.0 mL) was added to this slurry at the same temperature. The reaction mixture was then allowed to warm up to room temperature, and was quenched by addition of water (40.0 mL) . The precipitate was collected by filtration, washed with water and dried to afford the title compound as a gray solid (8.96 g, 44.0 mmol, 86% yield).

NMR Spectroscopy: ¾ NMR (700 MHz, (CD₃)₂SO, 25 °C, δ): 12.50 (s, 1H) , 8.44 (d, J = 3.2 Hz, 1H) , 8.16 (d, J = 7.6 Hz, 1H) , 7.55 (d, J = 7.7 Hz, 1H) , 7.25 - 7.33 (m, 2H) , 3.89 (s, 3H) . ¹³C NMR (175 MHz , (CD₃)2SO, 25 °C, δ) : 178.7, 164.0, 138.4, 136.7, 125.5, 123.8, 122.9, 121.1, 112.8, 112.4, 52.5. The ^lH NMR data were in good agreement with values reported in the literature (Ye, Q. et al. 2015) . Methyl 2- (5-bromo-lff-indol-3- l) -2-oxoacetate (5d)

To a solution of 5-bromo-lH-indole (l.OOg, 5.10 mmol, 1.00 equiv) in Et₂0 (6.00 mL, 0.850 M) was added oxalyl chloride (0.524 mL, 6.12 mmol, 1.20 equiv) dropwise at 0 °C. The yellow slurry was stirred at 0 °C for 30 min and then cooled to -78 °C. A solution of NaOMe in eOH (25%, 2.34 mL) was added to this slurry at the same temperature. The reaction mixture was then allowed to warm up to room temperature, and was quenched by addition of water (10.0 mL) . The precipitate was collected by filtration, washed with water and dried to afford the title compound as a gray solid (1.05 g, 3.72 mmol, 73% yield) .

NMR Spectroscopy: ^JH NMR (700 MHz, (CD3)₂SO, 25 °C, δ): 12.62 (s, 1H) , 8.51 (d, J = 2.2 Hz, 1H) , 8.29 (s, 1H) , 7.53 (d, J= 8.5 Hz, 1H) , 7.43 (d, J = 7.9 Hz, 1H), 3.89 (s, 3H) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ): 178.4, 163.4, 139.4, 135.5, 127.4, 126.5, 123.3, 115.6, 114.9, 111.9, 52.6. The ¹H NMR data were in good agreement with values reported in the literature (Ilovich, 0. et al . 2010) . Methyl 2- (l-ethyl-5-fluoro-lH-indol-3-yl) -2-oxoacetate (6a)

To a solution of methyl 2- ( 5-fluoro-lif-indol-3-yl ) -2-oxoacetate (1.23 g, 5.55 mmol, 1.00 equiv) in anhydrous DMF (20.0 mL, 0.278 M) was added sodium hydride (57-63% suspension in mineral oil, 289 mg, 7.22 mmol, 1.30 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 30 min before bromoethane (786 mg, 7.22 mmol, 1.30 equiv) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with ice water (60.0 mL) and the aqueous layer was extracted with EtOAc (6 ^χ 20.0 mL) . The organic extracts were combined, washed with 5% LiCl (aq) (5.00 mL) , and then dried over a₂ l¼ . After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel ( EtOAc : hexane3 = 1:2) to afford the title compound as a yellow solid (1.10 g, 4.41 mmol, 80% yield). NMR Spectroscopy: ^:H NMR (400 MHz, CDC1₃, 25 °C, δ): 8.43 (s, 1H) , 8.12 (dd, J = 2.6, 9.4 Hz, 1H) , 7.32 (dd, J = 4.2, 8.9 Hz, 1H) , 7.08 (dt, J = 2.6, 9.0 Hz, 1H) , 4.23 (q, J = 7.3 Hz, 2H) , 3.97 (s, 3H) , 1.58 (t, J = 7.3 Hz, 3H) . NMR (100 MHz, CDC1₃, 25 °C, δ) : 176.7,

163.3, 160.3 (d, J= 238.2 Hz), 139.7, 133.0, 128.3 (d, J= 27.3 Hz), 113.0, 112.5 (d, J = 26.3 Hz), 111.0 (d, J = 9.5 Hz), 108.6 (d, J = 25.1 Hz) , 52.9, 42.5, 15.1.

Methyl 2- -fluoro-l-methyl-lff-indol-3-yl) - -oxoacetate (6b)

To a solution of methyl 2- ( 5-fluoro-lii-indol-3-yl ) -2-oxoacetate (850 mg, 3.83 mmol, 1.00 equiv) in anhydrous DMF (10.0 mL, 0.383 M) was added sodium hydride (57-63% suspension in mineral oil, 169 mg, 4.20 mmol, 1.10 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 30 min before iodomethane (653 mg, 4.60 mmol, 1.20 equiv) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with ice water (30.0 mL) and the aqueous layer was extracted with EtOAc (4 ^χ 30.0 mL) . The organic extracts were combined, washed with 5% LiCl(aq) (5.00 mL) , and then dried over . After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:2) to afford the title compound as a white solid (860 mg, 3.66 mmol, 95 yield) . NMR Spectroscopy: ^]H NMR (400 MHz, CDCI3, 25 °C, δ): 8.28 (s, 1H) , 8.04 (dd, J = 2.4, 9.4 Hz, 1H) , 7.23 (dd, J = 4.2, 8.9 Hz, 1H) , 7.03 (dt, J = 2.5, 9.0 Hz, 1H) , 3.94 (s, 3H) , 3.82 (s, 3H) . ¹³C NMR (100 MHz , CDCI3, 25 °C, δ): 176.7, 163.2, 160.4 (d, J = 238.1 Hz), 141.4, 133.9, 128.0 (d, J = 11.2 Hz), 112.7 (d, J = 4.4 Hz), 112.5 (d, J = 26.3 Hz), 111.0 (d, J = 10.0 Hz), 108.2 (d, J = 25.2 Hz), 52.9, 34.2. The ¹H NMR ddLd were in good agreement with values reported in the literature (Ye, Q. eL al . 2015) .

Methyl 2- (5-fluoro-l-propyl-lH-indol-3-yl) -2-oxoacetate (6c)

To a solution of methyl 2- ( 5-fluoro-lH-indol-3-yl ) -2-oxoacetate (0.210 g, 0.949 mmol, 1.00 equiv) in anhydrous DMF (10.0 mL, 0.095 M) was added sodium hydride (57-63% suspension in mineral oil, 41.0 mg, 1.04 mmol, 1.10 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 30 min before 1-bromopropane (174 mg, 1.42 mmol, 1.50 equiv) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with ice water (40.0 mL) and the aqueous layer was extracted with EtOAc (4 ^χ 30.0 mL) . The organic extracts were combined, washed with 5% LiCl(aq) (5.00 mL) , and then dried over a₂ S0₄. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:2) to afford the title compound as a yellow solid (124 mg, 0.471 mmol, 50% yield) .

NMR Spectroscopy: ^JH NMR (400 MHz, CDCi;_:, 25 °C, δ): 8.41 (s, 1H) , 8.13 (dd, J = 2.6, 9.4 Hz, 1H), 7.32 (dd, J = 4.2, 8.9 Hz, 1H) , 7.08 (dt, J = 2.6, 9.0 Hz, 1H) , 4.15 (t, J = 7.2 Hz, 2H) , 3.96 (s, 3H) , 1.90-2.00 (m, 2H) , 0.99 (t, J = 7.4 Hz, 3H) . ^'-³C NMR (100 MHz, CDC13, 25 °C, δ ) : 176.8, 163.3, 160.4 (d, J = 237.9 Hz), 140.6, 133.3, 128.3 (d, J= 11.3 Hz), 112.9 (d, J= 4.3 Hz), 112.6 (d, J= 26.3 Hz), 111.2 (d, J = 9.6 Hz), 108.6 (d, J = 25.0 Hz), 53.0, 49.5, 23.3, 11.5.

Methyl 2- (l-butyl-5-fluoro-lH-indol-3-yl) -2-oxoacetate (6d)

To a solution of methyl 2- ( 5-fluoro-lff-indol-3-yl ) -2-oxoacetate (221 mg, 1.00 mmol, 1.00 equiv) in anhydrous DMF (10.0 mL, 0.100 M) was added sodium hydride (57-63% suspension in mineral oil, 44.0 mg, 1.10 mmol, 1.10 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 30 min before 1-bromobutane (206 mg, 1.50 mmol, 1.50 equiv) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with ice water (30.0 mL) and the aqueous layer was extracted with EtOAc (4 ^χ 30.0 mL) . The organic organic extracts were combined, washed with 5% LiCl(aq) (5.00 mL) , and then dried over Na_?SC>4. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:2) to afford the title compound as a white solid (200 mg, 0.721 mmol, 72% yield) .

NMR Spectroscopy: ^XH NMR (400 MHz, CDC1₃, 25 °C, δ) : 8.39 (s, 1H) , 8.10 (dd, J = 2.5, 9.4 Hz, 1H) , 7.29 (dd, J = 4.2, 8.9 Hz, 1H) , 7.06 (dt, J = 2.6, 9.0 Hz, 1H) , 4.15 (t, J = 7.2 Hz, 2H) , 3.95 (s, 3H) , 1.88 (q, J = 7.4 Hz, 2H) , 1.31-1.46 (m, 2H) , 0.97 (t, J = 7.4 Hz, 3H) . ¹³C NMR (100 MHz, CDC1₃, 25 °C, δ) : 176.7, 163.3, 160.3 (d, J = 238.2 Hz), 140.4, 133.2, 128.2 (d, J = 11.2 Hz), 113.0 (d, J = 4.3 Hz), 112.5 (d, J = 26.3 Hz), 111.1 (d, J = 9.8 Hz), 108.5 (d, J = 24.9 Hz), 52.9, 47.6, 31.8, 20.1, 13.7.

Methyl 2- (5-fluoro-l-isopropyl-lH-indol-3-yl) -2-oxoacetate (6e) (1.50 equiv)

0 "C to rt, overnight

51 %

To a solution of methyl 2- ( 5-fluoro-lH-indol-3-yl ) -2-oxoacetate (245 mg, 1.11 mmol, 1.00 equiv) in anhydrous DMF (5.00 mL, 0.222 M) was added sodium hydride (57-63% suspension in mineral oil, 48.8 mg, 1.22 mmol, 1.10 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 30 min before 2-bromopropane (283 mg, 1.66 mmol, 1.50 equiv) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with ice water (20.0 mL) and the aqueous layer was extracted with EtOAc (4 ^χ 20.0 mL) . The organic extracts were combined, washed with 5% LiCl(aq) (5.00 mL) , and then dried over Na₂S0₄. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel ( EtOAc : hexanes = 1:2) to afford the title compound as a white solid (150 mg, 0.570 mmol, 51% yield). NMR Spectroscopy: ^JH NMR (700 MHz, CDC1₃, 25 °C, δ) : 8.48 (s, 1H) , 8.10 (d, J= 7.0 Hz, 1H), 7.33 (t, J= 7.0 Hz, 1H) , 7.05 (s, lH) , 4.66 (t, J = 7.3 Hz, 2H) , 3.94 (s, 3H) , 1.60 (d, J = 6.7 Hz, 6H) . ¹³C NMR (175 MHz, CDCI3, 25 °C, δ): 176.7, 163.3, 160.2 (d, J = 238.1 Hz), 136.9, 132.9, 128.2 (d, J = 11.1 Hz), 113.1 (d, J = 4.2 Hz), 112.3 (d, J= 26.2 Hz), 111.2 (d, J= 9.7 Hz), 108.5 (d, J= 25.0 Hz), 52.9, 49.0, 22.6.

Methyl 2- (1- (2- (dime hylamino) ethyl) -5-fluoro-lff-indol-3-yl) -2- oxoacetate (6f)

To a solution of methyl 2- ( 5-fluoro-liT-indol-3-yl ) -2-oxoacetate (613 mg, 2.77 mmol, 1.00 equiv) in anhydrous DMF (15.0 mL, 0.185 M) was added sodium hydride (57-63% suspension in mineral oil, 288 mg, 7.2 mmol, 2.60 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 30 min before 2-chloro-N, W-dimethylethan-l-amine hydrochloride (479 mg, 3.32 mmol, 1.20 equiv) was added. The reaction mixture was heated to 50 °C and stirred overnight. The reaction was quenched with ice water (30.0 mL) and the aqueous layer was extracted with EtOAc (4 30.0 mL) . The organic extracts were combined, washed with 5% LiCl (aq) (5.00 mL) , and then dried over a2S0₄. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (CH₂Cl2:MeOH = 20:1) to afford the title compound as a colorless oil (410 mg, 1.40 mmol, 51% yield).

NMR Spectroscopy: ^XH NMR (700 MHz, CDCI3 , 25 °C, δ): 8.42 (s, 1H) , 8.05 (dd, J = 2.1, 9.4 Hz, 1H) , 7.28 (dd, J = 4.0, 8.8 Hz, 1H) , 7.02 (dt, J = 2.1, 8.8 Hz, 1H) , 4.21 (t, J - 6.7 Hz, 2H) , 3.92 (s, 3H) , 2.73 (t, J = 6.7 Hz, 2H) , 2.28 (s, 6H) . ¹³C NMR (175 MHz, CDCI3 , 25 °C, δ) : 176.8, 163.1, 160.2 (d, J = 237.7 Hz), 141.0, 133.2, 128.0 (d, J = 11.0 Hz), 112.9 (d, J= 3.9 Hz), 112.4 (d, J= 26.4 Hz), 111.0 (d, J = 9.9 Hz), 108.3 (d, J = 24.7 Hz), 58.3, 52.8, 45.7, 45.6.

Methyl 2- (5-fluoro-l- (2-morpholinoethyl) -lH-indol-3-yl) -2-oxoacetate (6g)

To a solution of methyl 2- ( 5-fluoro-lff-indol-3-yl) -2-oxoacetate (715 mg, 3.23 mmol, 1.00 equiv) in anhydrous DMF (15.0 mL, 0.215 M) was added sodium hydride (57-631 suspension in mineral oil, 155 mg, 3.88 mmol, 1.20 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 30 min before 4- (2-bromoethyl) morpholine (894 mg, 3.88 mmol, 1.20 equiv) was added. The reaction mixture was heated to 50 °C and stirred overnight. The reaction was quenched with ice water (30 mL) and the aqueous layer was extracted with EtOAc (4 ^χ 30.0 mL) . The organic extracts were combined, washed with 5% LiCl(aq) (5.00 mL) , and then dried over a2S0₄. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (CH₂Cl₂:MeOH = 10:1) to afford the title compound as a colorless oil (460 mg, 1.38 mmol, 43% yield) .

NMR Spectroscopy: ^l NMR (700 MHz, CDCI₃, 25 °C, δ) : 8.47 (s, 1H) , 8.06 (dd, J = 2.4, 9.3 Hz, 1H) , 7.29 (dd, J = 4.0, 8.8 Hz, 1H) , 7.04 (dd, J = 2.5, 8.8 Hz, 1H) , 4.24 (t, J = 6. 1 Hz, 2H) , 3.93 ( s , 3H) , 3.68 (d, J = 3.9 Hz, 4H) , 2.78 (t, J = 6.1 Hz, 2H) , 2.49 (s, 4H) . ¹³C NMR (175 MHz, CDCI₃, 25 °C, δ) : 176.8, 163. 1 , 160.3 (d, J = 238 Hz), 141.3, 133.2, 128.0 (d, J = 11.3 Hz), 112.9 (d, J = 4.1 Hz), 112.4 (d, J = 26.3 Hz), 111.0 (d, J= 9.9 Hz), 108.4 (d, J = 24.6 Hz), 67.0, 57.5, 53.8, 52.9, 44.9.

Methyl 2- <5-fluoro-1- (2-me hoxyethyl) -lH-indol-3-yl) -2-oxoacetate (6h)

To a solution of methyl 2- ( 5-fluoro-lH-indol-3-yl ) -2-oxoacetate (570 mg, 2.58 mmol, 1.00 equiv) in anhydrous DMF (15.0 mL, 0.172 M) was added sodium hydride (57-63% suspension in mineral oil, 114 mg, 2.84 mmol, 1.10 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 30 min before 1 -bromo-2 -methoxyethane (430 mg, 3.09 mmol, 1.20 equiv) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with ice water (30.0 mL) and the aqueous layer was extracted with EtOAc (4 ^χ 30.0 mL) . The organic extracts were combined, washed with 5% LiCl(aq) (5.00 mL) , and then dried over NaiSC . After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel ( EtOAc : hexanes = 1:1) to afford the title compound as a colorless oil (401 mg, 1.44 mmol, 56% yield) .

NMR Spectroscopy: NMR (700 MHz, CDCl₃, 25 °C, δ): 8.33 (s, 1H),

7.80-8.06 (m, III), 7.10 7.28 (m, 1H) , 6.82-7.03 (m, 1H) , 4.2? (s, 2H), 3.88 (s, 3H), 3.66 (s, 2H) , 3.24 (s, 3H) . ¹³C NMR (175 MHz, CDCI3, 25 "C, δ) : 176.8, 163.0, 160.0 (d, J= 237.4 Hz), 141.3, 133.1, 127.6 (d, J = 10.4 Hz), 112.6, 112.0 (d, J = 26.1 Hz), 111.2 (d, J = 8.4 Hz), 107.8 (d, J = 24.9 Hz), 70.3, 58.8, 52.6, 47.0.

Methyl 2- (l-

To a solution of methyl 2- ( lH-indol-3-yl ) -2-oxoacetate (1.04 g, 5.12 mmol, 1.00 equiv) in anhydrous DMF (25.0 mL, 0.205 M) was added sodium hydride (57-63% suspension in mineral oil, 225 mg, 5.63 mmol, 1.10 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 30 min before iodomethane (872 mg, 6.14 mmol, 1.20 equiv) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with ice water (50.0 mL) and the aqueous layer was extracted with EtOAc (6 ^χ 30.0 mL) . The organic extracts were combined, washed with 5% LiCl(aq) (5.00 mL) , and then dried over Na S0₄ . After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:1) to afford the title compound as a white solid (1.01 g, 4.65 mmol, 91% yield) .

NMR Spectroscopy: ^:H NMR (400 MHz, CDCl₃, 25 °C, δ): 8.40-8.46 (m, 1H) , 8.29 (s, 1H), 7.31- 7.40 (m, 3H) , 3.95 (s, 3H) , 3.84 (s, 3H) . ¹³C NMR (100 MH z , CDCI3, 25 °C, δ) : 177.0, 163.5, 140.6, 137.5, 127.1, 124.3, 123.7, 122.8, 112.9, 110.1, 52.8, 33.9. The ^lH NMR data were in good agreement with values reported in the literature (M. Wang et al . 2011) . Methyl 2- (5-chloro-l-methyl-lfl-indol-3-yl) -2-oxoacetate (6j)

5b 85%

To a solution of methyl 2- (5-chloro-lii-indol-3-yl) -2-oxoacetate (1.00 g, 4.20 mmol, 1.00 equiv) in anhydrous DMF (15.0 mL, 0.280 M) was added sodium hydride (57-63% suspension in mineral oil, 185 mg, 4.62 mmol, 1.10 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 30 min before iodomethane (715 mg, 5.04 mmol, 1.20 equiv) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with ice water (30.0 mL) and the aqueous layer was extracted with EtOAc (5 ^χ 30.0 mL) . The organic extracts were combined, washed with 5% LiCl(aq) (5.00 mL) , and then dried over Na2≤04. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:1) to afford the title compound as a white solid (900 mg, 3.58 mmol, 85% yield) . NMR Spectroscopy: ¾ NMR (500 MHz, CDCI3, 25 °C, δ) : 8.27 (d, J = 1.9 Hz, 1H) , 8.21 (s, 1H), 7.13-7.20 (m, 2H) , 3.93 (s, 3H) , 3.79 (s, 3H) . ¹³C NMR (125 MHz, CDCI3, 25 °C, δ): 176.6, 163.0, 141.1, 135.6, 129.4, 127.9, 124.2, 121.9, 112.2, 111.1, 52.9, 34.0. The 1H NMR data were in good agreement with values reported in the literature (Ye, Q. et al. 2015) .

Methyl 2- (5-bromo-l-meth l-lH-indol-3-yl) -2-oxoacetate 6k)

To a solution of methyl 2- (5-bromo-li7-indol-3-yl ) -2-oxoacetate (984 _mg, 3.49 mmol, 1.00 equiv) in anhydrous DMF (12.0 mL, 0.291 M) was added sodium hydride (57-63% suspension in mineral oil, 168 mg, 4.19 mmol, 1.20 equiv) at 0 °c. The reaction mixture was stirred at 0 °C for 30 min before iodomethane (595 mg, 4.19 mmol, 1.20 equiv) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with ice water (30.0 rtiL) and the aqueous layer was extracted with EtOAc (5 ^χ 30.0 iriL) . The organic extracts were combined, washed with 5% LiCl(aq) (5.00 mL) , and then dried over a2S0 . After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:1) to afford the title compound as a white solid (739 mg, 2.50 mmol, 72% yield) .

NMR Spectroscopy: ^lH NMR (700 MHz, CDC1₃, 25 °C, δ) : 8.34 (s, 1H) , 8.11 (s, 1H), 7.25 (d, J = 7.7 Hz, 1 H) , 7.03 (d, J = 8.1 Hz, 1 H) , 3.91 (s, 3H), 3.74 (s, 3H) . ¹³C NMR (175 MHz, CDC1₃, 25 °C, δ) : 176.5, 162.9, 141.0, 135.8, 128.3, 126.9, 124.8, 117.1, 112.0, 111.5, 52.9, 34.0. The ^XH NMR data were in good agreement with values reported in the literature (Q. Ye et al 2015).

2- (3-Hydroxy henyl) acetamide (8)

To a solution of 2- ( 3-hydroxyphenyl ) acetic acid (20.0 g, 130 mmol, 1.00 equiv) in MeOH (300 mL, 0.433 M) was added concentrated sulphuric acid (1.00 mL) . The solution was heated to reflux for 5 h. The solvent was evaporated and the residue was dissolved in CH2CI2 (300 mL) and washed with water (3 ^χ 20.0 mL) . The organic layer was dried over Na:SC¼ and evaporated to dryness to afford an ester. To the residue was added 28-30% NH₃ -H20 (110 mL) at 0 °C, then the mixture was stirred overnight at room temperature. The resulting solution was evaporated and the residue neutralized with acetic acid. The crude product was recrystal 1 i zed from EtOAc to afford the title compound as a white crystalline solid (10.8 g, 71.4 mmol, 55% yield).

NMR Spectroscopy: ^JH NMR (400 MHz, (CD₃)₂SO, 25 °C, δ): 9.27 (s, 1H), 7.39 (s, 1H) , 7.06 (t, J = 7.8 Hz, 1H) , 6.83 (s, 1H) , 6.50-6.72 (m, 3H) , 3.26 (s, 2H) . ¹³C NMR (100 MHz, (CD₃)₂SO, 25 °C, δ): 172.2, 157.2, 137.7, 129.0, 119.7, 115.9, 113.2, 42.3. All the analytical data were in good agreement with values reported in the literature (WSS Spectral Data from Wiley) .

2- (3- (2-Fluoroethoxy) phenyl) acetamide (9)

To a solution of 2- (3-hydroxyphenyl) acetamide (466 mg, 3.09 mmol, 1.00 equiv) in DMF (20.0 mL, 0.155 M) was added K2C03 (2.56 g, 18.5 mmol, 6.00 equiv), Nal (93.0 mg, 0.62 mmol, 0.200 equiv), and l-bromo-2- fluoroethane (510 mg, 4.01 mmol, 1.30 equiv) under nitrogen atmosphere at room temperature. The reaction mixture was heated to 60 °C and stirred for 20 h. The reaction was cooled to 0 °C and quenched with water (50.0 mL) and the aqueous layer was extracted with EtOAc (6 ^χ 30.0 mL) . The organic extracts were combined, washed with 5% LiCl (aq) (5.00 mL) , and then dried over a₂S0₄. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc) to afford the title compound as a white solid (494 mg, 2.50 mmol, 81% yield) .

NMR Spectroscopy: ^]H NMR (400 MHz, (CD₃)₂SO, 25 °C, δ) : 7.44 (s, 1H) , 7.21 (t, J = 7.8 Hz, 1H) , 6.75-6.91 (m, 4H) , 4.73 (td, J = 3.9, 47.8 Hz, 2H), 4.20 (td, J = 3.9, 30.1 Hz, 2H) , 3.36 (s, 2H) . ¹³C NMR (100 MHz, (CD₃);-SO, 25 °C, 5): 172.1, 158.0, 138.1, 129.2, 121.7, 115.5, 112.2, 82.2 (d, J = 165.7 Hz), 66.9 (d, J = 18.8 Hz), 42.3.

3- (l-Ethyl-5-fluoro-lH-indol-3-yl) -4- (3-me hoxyphenyl) -lH-pyrrole- 2,5-dione (10a)

Under N₂ atmosphere, to a suspension of methyl 2- ( l-ethyl-5-fluoro- lif-indol-3-yl) -2~oxoacetate (410 mg, 1.65 mmol, 1.00 equiv) and 2-(3- ( 2-fluoroethoxy) phenyl ) acetamide (357 mg, 1.81 mmol, 1.10 equiv) in THF (10.0 mL, 0.165 M) was slowly added (dropwise) t-BuOK solution (463 mg in 20.0 mL THF, 4.13 mmol, 2.5 mmol) at 0 °C for 30 min. The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with HCl (IN, 5.0 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 ^χ 20.0 mL) . The organic extracts were combined, washed with saturated NaCl (aq) (5.00 mL) , and then dried over Na₂S0₄. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAchexanes = 1:1) to afford the title compound as a red solid (450 mg, 1.14 mmol, 69% yield).

NMR Spectroscopy: ¾ NMR (400 MHz, (CD₃)₂SO, 25 °C, δ): 11.10 (s, 1H) , 8.13 (s, 1H), 7.56 (dd, ,7= 4.6, 9.0 Hz, 1H) , 7.20-7.31 (m, 1H) , 6.90- 7.05 (m, 4H) , 5.98 (dd, J = 2.5, 10.6 Hz, 1H) , 4.62-4.70 (m, 1H) , 4.51-4.58 (m, lH) , 4.32 (q, J = 7.2 Hz, 2H), 4.00-4.15 (m, 2 H) , 1.41 (t, J = 7.2 Hz, 3H) . ¹³C NMR (100 MHz, (CD₃)₂SO, 25 °C, δ): 172.2, 172.0, 157.7, 156.9 (d, J = 232.6 Hz), 134.8, 132.7, 132.0, 131.7, 129.3, 128.5, 125.1 (d, J = 10.5 Hz), 122.5, 116.0, 115.0, 111.8 (d, J = 9.8 Hz), 110.1 (d, J = 25.7 Hz), 106.5 id, J = 25.0 Hz), 103.3 (d, J = 4.1 Hz), 81.9 (d, J= 165.9 Hz), 67.1 (d, J = 19.1 Hz), 41.2, 15.2. ¹⁹F NMR (376 MHz, (CD₃)2SO, 25 °C, δ): -124.6 (m) , - 224.5 (m) . HRMS (ESI-TOF) (m/z): calcd for C22H19F2N2O3 ( [M + H]⁺), 397.1364, found, 397.1357.

3- (5-Fl oro-l-met yl-lH-indol-3-yl) -4- (3- (2-fluoroethoxy) henyl) -lff- pyrrole-2 , 5-dione (10b)

Under ₂ atmosphere, to a suspension of methyl methyl 2- (b-fluoro-1- methyl-lfi-indol-3-yl ) -2- oxoacetate (316 mg, 1.34 mmol, 1.00 equiv) and 2- (3- ( 2-fluoroethoxy) phenyl) acetamide (318 mg, 1.61 mmol, 1.20 equiv) in THF (15.0 mL, 0.089 M) was slowly added (dropwise) t-BuOK solution (451 mg, 4.02 mmol in 15 mL THF, 3.00 equiv) at 0 °C for 30 min. The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with HC1 (I , 5.00 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 10.0 mL) . The organic extracts were combined, washed with saturated NaCl (aq) (5.00 mL) , and then dried over Na₂S0₄. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:1) to afford the title compound as a red solid (240 mg, 0.628 mmol, 47% yield) .

NMR Spectroscopy: ¾ NMR (400 MHz, (CD₃)₂SO, 25 °C, δ): 11.09 (s, IH) , 8.10 (s, IH) , 7.51 (dd, J = 4.6, 9.0 Hz, IH) , 7.26 (t, J = 7.8 Hz, IH) , 6.97-7.04 (m, 3H) , 6.95 (d, J = 7.7 Hz, IH) , 5.94 (dd, J = 2.4, 10.6 Hz, IH) , 4.63 (td, J= 3.7, 47.9 Hz, 2H) , 4.09 (td, J= 3.7, 30.0 Hz, 2H) , 3.90 (s, 3H) . ¹³C NMR (175 MHz, (CD3J2SO, 25 °C, δ) : 172.2, 172.1, 157.7, 157.0 (d, J = 233.7 Hz), 136.4, 133.7, 131.9, 131.8, 129.4, 128.4, 124.9, 122.5, 116.0, 115.0, 111.8 (d, J = 10.0 Hz), 110.1 (d, J = 25.8 Hz), 106.4 (d, J = 25.1 Hz), 103.1, 81.9 (d, J = 165.6 Hz), 67.1 (d, J= 18.9 Hz), 33.3. ¹⁹F NMR (376 MHz, (CD₃)₂SO, 25 °C, δ) : -124.5 (m) , -224.4 (m) . HRMS (ESI-TOF) (m/z): calcd for C₂iHi₇F_{2 2}0₃ ( [M + H]⁺), 383.1207, found, 383.1202.

3- (5-Fluoro-l-propyl-lH-indol-3-yl) -4- (3- (2-fluoroethoxy) henyl) -1H- pyrrole-2 , 5-dione (10c)

Under N₂ atmosphere, to a suspension of methyl methyl 2- (5-fluoro-1- propyl-lH-indol-3-yl) -2- oxoacetate (60.0 mg, 0.228 mmol, 1.00 equiv) and 2- (3- ( 2-fluoroethoxy) phenyl) acetamide (54.0 mg, 0.274 mmol, 1.20 equiv) in THF (5.00 mL) was added t-BuOK solution (64.0 mg, 0.570 mmol in 10.0 mL THF, 2.50equiv) at 0 °C for 30 min. The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with HC1 (IN, 5.00 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 x 10.0 mL) . The organic extracts were combined, washed with saturated NaCl (aq) (5 mL) , and then dried over a2S0₄. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:1) to afford the title compound as a red solid (56.1 mg, 0.134 mmol, 60% yield).

NMR Spectroscopy: ¾ NMR (700 MHz, ((CD₃)₂SO, 25 °C, δ) : 11.10 (s, 1H) , 8.10 (s, 1H), 7.57 (t, J = 4.4 Hz, 1H) , 7.27 (t, J = 7.9 Hz, 1H) , 6.90-7.04 (m, 4H) , 6.01 (dd, J= 2.1, 10.4 Hz, 1H) , 4.61 (td, J= 3.5, 47.9 Hz, 2H) , 4.25 (t, J = 7.0 Hz, 2H) , 4.06 (td, J = 3.5, 30.0 Hz, 2H) , 1.80 (q, J = 7.1 Hz, 2H) , 0.85 (t, J = 7.3 Hz, 3H) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ): 172.1, 172.0, 157.7, 157.0 (d, J = 232.6 Hz), 135.5, 133.0, 132.0, 131.7, 129.4, 128.6, 125.1 (d, J= 10.7 Hz), 122.4, 115.9, 115.1, 111.9 (d, J = 9.8 Hz), 110.1 (d, J = 25.6 Hz), 106.5 (d, J= 25.0 Hz), 103.2 (d, J= 4.1 Hz), 81.9 (d, J= 165.7 Hz), 67.0 (d, J = 18.9 Hz), 47.8, 23.0, 11.0. ^,9F NMR (376 MHz, (CD₃)2SO, 25 °C, δ) : -124.5 (m) , -224.6 (m) . HRMS (ESI-TOF) (m/z): calcd for C₂3H2iF₂ 203 ( [M + Hp), 411.1520, found, 411.1534.

3- (l-Butyl-5-fluoro-lH-indol-3-yl) -4- (3- (2-fluoroethoxy) phenyl) -1H- pyrrole-2 , 5-dione (10d)

Under 2 atmosphere, to a suspension of methyl methyl 2- ( l-butyl-5- fluoro-lH-indol-3-yl) -2- oxoacetate (106 mg, 0.380 mmol, 1.00 equiv) and 2- (3- (2-fluoroethoxy) phenyl ) acetamide (90.5 mg, 0.459 mmol, 1.20 equiv) in THF (10.0 mL) was slowly added (dropwise) t-BuOK solution (102 mg, 0.912 mmol in 10 mL THF, 2.40 equiv) at 0 °C for 30 min. The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with HCl (IN, 5.00 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 ^χ 10.0 mL) . The organic extracts were combined, washed with saturated NaCl aqueous (5.00 mL) , and then dried over Na2S0 . After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1/3) to afford the title compound as a red solid (100 mg, 0.236 mmol, 62% yield) . NMR Spectroscopy: H NMR (400 MHz, (CD₃)₂SO, 25 °C, δ): 11.11 (s, 1H) , 8.08 (s, 1H), 7.54 (dd, J = 4.6, 9.0 Hz, 1H) , 7.22-7.30 (m, 1H) , 6.90- 7.03 (m, 4H), 6.03 (dd, J= 2.4, 10.6 Hz, 1H) , 4.61 (td, J = 3.7, 47.9 Hz, 2H) , 4.26 (t, J = 7.0 Hz, 2H) , 4.06 (td, J = 3.7, 30.0 Hz, 2H) , 1.70-1.79 (m, 2H) , 1.22-1.32 (m, 2H) , 0.88 (t, J = 7.4 Hz, 3H) . ¹³C NMR (100 MHz, (CD₃).-SO, 25 °C, δ): 172.2, 172.0, 157.7, 157.0 (d, J = 232.6 Hz), 135.3, 132.9, 132.0, 131.7, 129.3, 128.6, 125.1 (d, J =

10.7 Hz), 122.5, 115.9, 115.1, 111.8 (d, J = 9.7 Hz), 110.1 (d, J =

25.8 Hz), 106.5 (d, J = 24.9 Hz), 103.3 (d, J = 4.2 Hz), 81.9 (d, J = 166.1 Hz), 67.0 (d, J = 19.0 Hz), 64.9, 46.0, 31.6, 19.4, 13.5. ¹⁹F NMR (376 MHz, (CD₃)₂SO, 25 °C, δ): -121.6 (m) , -223.2 (m) . HRMS (ESI- TOF) (m/z): calcd for

([M + H]⁺), 425.1677, found, 425.1679.

3- (5-Fluoro-l-isopropyl-lH-indol-3-yl) -4- (3- (2-fluoroethoxy) henyl) - lH-pyrrole-2 , 5-dione (lOe)

Under 2 atmosphere, to a suspension of methyl methyl 2- (5-fluoro-l- isopropyl-lff-indol-3-yl) -2- oxoacetate (90.0 mg, 0.342 mmol, 1.00 equiv) and 2- ( 3- ( 2-fluoroethoxy) phenyl ) acetamide (80.5 mg, 0.408 mmol, 1.19 equiv) in THF (5.00 mL, 0.068 M) was added t-BuOK solution (95.9 mg, 0.855 mmol in 10 mL THF, 2.50 equiv) at 0 °C for 30 min. The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with HC1 (IN, 5.00 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 * 10.0 mL) . The organic extracts were combined, washed with saturated NaCl (aq) (5.00 mL) , and then dried over Na₂S0 . After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel ( EtOAc : hexanes = 1:2) to afford the title compound as a red solid (91.0 mg, 0.223 mmol, 65% yield).

NMR Spectroscopy: ^JH NMR (400 MHz, (CD₃)₂SO, 25 °C, δ) : 11.11 (s, 1H) , 8.13 (s, 1H), 7.61 (dd, J= 4.6, 9.0 Hz, 1H) , 7.22-7.31 (m, 1H) , 6.90- 7.04 (m, 4H) , 6.04 (dd, J = 2.5, 10.6 Hz, 1H) , 4.85 (q, J = 6.7 Hz, 1H), 4.62 (td, J =3.8, 51.4 Hz, 2H) , 4.09 (td, J= 3.8, 30.0 Hz, 2H) , 1.50 (d, J = 6.7 Hz, 6H) . ¹³C NMR (100 MHz, (CD₃)₂SO, 25 °C, δ): 172.1, 172.0, 157.7, 157.0 (d, J = 233.0 Hz), 132.5, 132.0, 131.7, 131.5, 129.4, 128.6, 125.1 (d, J = 10.4 Hz), 122.4, 115.9, 115.0, 111.9 (d, J = 8.6 Hz),, 110.1 (d, J = 24.1 Hz),, 106.6 (d, J = 22.6 Hz), 103.7, 81.9 (d, J = 165.8 Hz), 67.1 (d, J = 18.8 Hz), 47.7, 22.3. ¹⁹F NMR (376 MHz , (CD₃)₂SO, 25 °C, δ): -124.4 (m) , -224.4 (m) . HRMS (ESI-TOF) (m/z) : calcd for C?3H₂:F₂N₂0₃ ( [M + H]⁺), 411.1520, found, 411.1534.

3- (1- (2- (Dimethylamino) ethyl) -5-fluoro-lH-indol-3-yl) -4- (3- (2- fluoroethoxy) henyl) -lH-pyrrole-2 , 5-dione (lOf)

if

Under 2 atmosphere, to a suspension of methyl 2-(l-(2- (dimethylamino) ethyl ) -5-fluoro-lif-indol- 3-yl ) -2-oxoacetate (318 mg, 1.09 mmol, 1.00 equiv) and 2- (3- (2-fluoroethoxy) phenyl) acetamide (257 mg, 1.31 mmol, 1.20 equiv) in THF (20.0 mL, 0.055 M) was slowly added (dropwise) t-BuOK solution (367 mg, 3.27 mmol in 15 mL THF, 3.00 equiv) at 0 °C for 30 min . The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with ice water (5.00 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 ^χ 20.0 mL) . The organic extracts were combined, washed with saturated NaCl (aq) (5.00 mL) , and then dried over Na2S04. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (CH₂Cl₂:MeOH = 20:1) to afford the title compound as a yellow solid (330 mg, 0.751 mmol, 69% yield).

NMR Spectroscopy: ²H NMR (700 MHz, (CD₃)₂SO, 25 °C, δ) : 11.10 (s, 1H) , 8.13 (s, 1H), 7.57 (t, J = 4.4 Hz, 1H) , 7.28 (t, J = 7.9 Hz, 1H) , 6.90-7.10 (m, 4H) , 6.02 (dd, J= 2.5, 10.5 Hz, 1H) , 4.61 (td, J= 3.8, 47.8 Hz, 2H) , 4.37 (t, J = 6.4 Hz, 2H), 4.06 (td, J = 3.8, 30.0 Hz, 2H) , 2.65 (t, J = 6.3 Hz, 2H) , 2.19 (s, 6H) . "C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ) : 172.2, 172.0, 157.7, 157.0 (d, J= 232.5 Hz), 156.3, 135.8, 133.1, 132.0, 131.7, 129.4, 128.6, 125.0 (d, J = 10.3 Hz), 122.4, 115.8, 115.1, 111.8 (d, J = 9.9 Hz), 110.1 (d, J = 25.6 Hz), 106.5 (d, J = 24.8 Hz), 103.3 (d, J = 3.8 Hz), 81.9 (d, J = 165.8 Hz), 67.0 (d, J = 19.0 Hz), 58.3, 45.2, 44.3. ¹⁹F NMR (376 MHz, (CD₃)₂SO, 25 °C, δ) : -124.7 (m) , -224.4 (m) . HRMS (ESI-TOF) (m/z): calcd for C₂-iH2₄F₂N₃03 ( [M + H]'), 440.1786, found, 440.1785.

3- (5-Fluoro-l- (2-morpholinoethyl) -lff-indol-3-yl) -4" (3- (2- fluoroethoxy) phenyl) -lH-pyrrole-2 , 5-dione (lOg)

Under 2 atmosphere, to a suspension of methyl 2 - (5-fluoro-l- ( 2 - morpholinoethyl ) -lH-indol- 3 - yl ) -2-oxoacetate ( 360 mg, 1. 08 mmol, 1.00 equiv) and 2- ( 3 - (2-fluoroethoxy) phenyl) acetamide (255 mg, 1 . 30 mmol, 1.20 equiv) in THF (20 mL, 0.054 M ) was slowly added (dropwise) t-BuOK solution (364 mg, 3.24 mmol in 15 mL THF, 3.00 equiv) at 0 °C for 30 min. The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with ice water ( 5 . 00 mL) at 0°C and the aqueous layer was extracted with EtOAc (4 20 . 0 mL) . The organic extracts were combined, washed with saturated NaCl (aq) ( 5 mL) , and then dried over Na2S04. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (CH₂Cl₂:MeOH = 20 : 1 ) to afford the title compound as a yellow solid (360 mg, 0. 748 mmol, 69% yield). N R Spectroscopy: ²H NMR (700 MHz, (CD₃)₂SO, 25 °C, δ) : 11.10 (s, 1H), 8.18 (s, 1H) , 7.57 (t, J = 4.4 Hz, 1H) , 7.27 (t, J = 7.9 Hz, 1H) , 6.90-7.04 (m, 4H) , 5.99 (dd, J = 2.2, 10.4 Hz, 1H) , 4.61 (td, J = 3.5, 47.9 Hz, 2H) , 4.38 (t, J = 6.3 Hz, 2H) , 4.07 (td, J = 3.5, 30.0 Hz, 2H), 3.54 (s, 4H) , 2.68 (t, J = 6.2 Hz, 2H) , 2.42 (s, 4H) . ¹³C NMR (100 MHz, (CD₃)?SO, 25 °C, δ): 172.2, 172.0, 157.7, 157.0 (d, J = 232.5 Hz), 136.1, 133.1, 132.0, 131.8, 129.4, 128.5, 125.0 (d, J = 10.3 Hz), 122.5, 116.0, 115.0, 111. 8 (d, J = 9.8 Hz), 110.0 (d, J= 25.9 Hz), 106.4 (d, J = 25.1 Hz), 103.3 (d, J= 4.3 Hz) , 81.9 (d, J = 165.7 Hz), 67.1 (d, J = 18.9 Hz), 66.2, 57.3, 53.2, 43.5. ¹³F NMR (376 MHz, (CD₃)_zSO, 25 °C, δ): -124.7 (m) , -224.4 (m) . HRMS (ESI- TOF ) (m/z) : calcd for C₂₆H_2fcF_N₃0₄ ([M + H]⁺), 482.1891, found, 482.1883.

3- (5-Fluoro-l- (2-methoxyethyl) -lff-indol-3-yl) -4- (3- (2-fluoroethoxy) phenyl) -lii-pyrrole-2 , 5- dione (lOh)

Under N₂ atmosphere, to a suspension of methyl 2 - (5-fluoro-l- ( 2 - methoxyethyl ) -lff-indol-3-yl ) - 2-oxoacetate (370 mg, 1.32 mmol, 1.00 equiv) and 2 - ( 3- (2-fluoroethoxy) phenyl ) acetamide (312 mg, 1.58 mmol, 1.20 equiv) in THF (20 mL, 0. 066 M) was slowly added (dropwise) t- BuOK solution (415 mg, 3.70 mmol in 15 mL THF, 2 . 80 equiv) at 0 °C for 30 min. The reaction mixture was stirred at 0 ° C for another 4 h. The reaction was quenched with ice water (5.00 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 ^χ 20.0 mL) . The organic extracts were combined, washed with saturated NaCl (aq) ( 5.00 mL) , and then dried over Na₂S0₄. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel ( EtOAc : hexanes = 1:2) to afford the title compound as a red fluffy solid (197 mg, 0.462 mmol, 35% yield). NMR Spectroscopy: ^lH NMR (700 MHz, (CD₃)₂SO, 25 °C, δ ) : 11.10 (s, 1H) , 8 . 10 (s, 1H) , 7.58 (t, J = 4.4 Hz, 1H), 7.28 (t, J = 7.9 Hz, 1H) , 6.90-7.05 (m, 4H) , 6.01 (dd, J = 1.8, 10.4 Hz, 1H) , 4.62 (td, J = 3.3, 51.1 Hz, 2H) , 4.45 (t, J = 4.9 Hz, 2H) , 4.07 (td, J = 3.3, 30.0 Hz, 2H) , 3.69 (t, J = 4.9 Hz, 2H) , 3.23 (s, 3H) . ¹³C MR (175 MHz, (CD₃)₂SO, 25 °C, δ) : 172.2, 172.0, 157.7, 157.0 (d, J = 232.5 Hz), 156.3, 135.8, 133.2, 131.9, 131.7, 129.4, 128.6, 125.0 (d, J = 10.9 Hz), 122.5, 115.9, 115.1, 112.0 (d, J = 9.7 Hz), 110.1 (d, J = 25.6 Hz), 106.4 (d, J = 25.1 Hz), 103.4 (d, J= 4.3 Hz), 81.9 (d, J = 165.7 Hz), 70.7, 67.0 (d, J = 19.0 Hz), 58.1, 46.2. ¹⁹F NMR (376 MHz, (CD₃)₂SO, 25 °C, δ): -124.6 (m) , -224.4 (m) . HRMS (ESI-TOF) (m/z): calcd for C₂₃H₂iF_{? 2}0₄ ( [M + H]⁺), 427.1469, found, 427.1463.

3- (3- (2-Fluoroethoxy) henyl) -4- ( l-methyl-l/i-indol-3-yl) -lH-pyrrole- 2,5-dione (lOi)

6i 9 101

Under N2 atmosphere, to a suspension of methyl 2- ( l-methyl-lH-indol- 3-yl) -2-oxoacetate (333 mg, 1.53 mmol, 1.00 equiv) and 2-(3-(2- fluoroethoxy) phenyl ) acetamide (333 mg, 1.69 mmol, 1.10 quiv) in THF (10.0 mL, 0.153 M) was slowly added (dropwise) t-BuOK solution (516 mg, 4.60 mmol in 25 mL THF, 3.00 equiv) at 0 °C for 30 min. The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with HC1 (IN, 10.0 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 ^χ 20.0 mL) . The organic extracts were combined, washed with saturated NaCl (aq) (5 mL) , and then dried over a2SOi . After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:2) to afford the title compound as a red solid (280 mg, 0.768 mmol, 50% yield). NMR Spectroscopy: ¾ NMR (700 MHz, (CD₃)₂SO, 25 °C, δ): 11.08 (s, 1H) , 8.04 (s, 1H) , 7.49 (d, J = 8.2 Hz, 1H) , 7.22 (t, J= 7.6 Hz, 1H) , 7.00 (s, 1H) , 6.90-6.98 (m, 2H) , 6.76 (t, J = 7.6 Hz, 1H) , 6.34 (d, J = 8.1 Hz, 1H), 4.60 (td, J = 3.7, 47.8 Hz, 2H) , 4.04 (td, J= 3.7, 30.1 Hz, 2H) , 3.90 (s, 3H) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ ) : 172.4, 172.2, 157.6, 137.1, 134.9, 132.2, 131.9, 129.2, 128.0, 124.4, 122.5, 122.1, 120.1, 115.9, 114.9, 110.6, 103.1, 81.9 (d, J = 165.6), 67.0 (d, J = 18.9), 33.1. ¹⁹F NMR (376 MHz, (CD₃)₂SO, 25 °C, δ): -224.4 (m) . HRMS (ESI-TOF) (m/z): calcd for C21H18FN2O3 ( [M + H]⁺), 365.1301, found, 365.1289. 3- (5-Chloro-l-methyl-lH-indol-3-yl) -4- (3- (2-fluoroethoxy) henyl) -1H- pyrrole-2 , 5-dione (10 )

10J

Under N₂ atmosphere, to a suspension of methyl 2- ( 5-chloro-l-methyl- lH-indol-3-yl) -2- oxoacetate (279 mg, 1.11 mmol, 1.00 equiv) and 2- (3- (2-fluoroethoxy) phenyl) acetamide (262 mg, 1.33 mmol, 1.20 equiv) in THF (20 mL, 0.056 M) was slowly added (dropwise) t-BuOK solution (374 mg, 3.33 mmol in 15 mL THF, 3.00 equiv) at 0 °C for 30 min. The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with ice water (5.00 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 ^χ 20.0 mL) . The organic extracts were combined, washed with saturated NaCl (aq) (5.00 mL) , and then dried over a2SO« . After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:2) to afford the title compound as a yellow solid (270 mg, 0.677 mmol, 61% yield). N R Spectroscopy: ^]H N R (700 MHz, (CDahSO, 25 °C, δ): 11.11 (s, 1H) , 8.10 (s, 1H) , 7.52 (d, J = 8.6 Hz, 1H) , 7.28 (t, J = 7.9 Hz, 1H), 7.14 (dd, J = 1.8, 8.6 Hz, 1H) , 7.03 (dd, J = 2.3, 8.3 Hz, 1H), 6.98 (s, 1H), 6.94 (d, J = 7.6 Hz, 1H) , 6.21 (d, J= 1.8 Hz, 1H), 4.62 (td, J= 3.8, 47.8 Hz, 2H) , 4.08 (td, J= 3.8, 30.0 Hz, 2H) , 3.90 (s, 3H) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ): 172.2, 172.0, 157.8, 136.1, 135.5, 131.8, 129.3, 128.8, 125.5, 124.9, 122.6, 121.9, 121.0, 116.2, 115.1, 112.0, 102.9, 81.9 (d, J = 166.1 Hz), 67.2 (d, J = 19.0 Hz), 20.8. ¹⁹F NMR (376 MHz, (CD₃)₂SO, 25 °C, δ) : -224.4 (m) . HRMS (ESI-TOF) (m/z): calcd for C₂1H1₇CI FN₂O3 ( [M + H]⁺), 399.0912, found, 399.0900.

3- ( 5 -Bromo-l -methyl- lH-indol-3-yl) -4 - ( 3- ( 2- luoroethoxy) phenyl ) - 1H- pyrrole-2 , 5-dione ( 10k)

9 10k

Under 2 atmosphere, to a suspension of methyl 2- (5-bromo-l-methyl- lH-indol-3-yl) -2- oxoacetate (120 mg, 0.405 mmol, 1.00 equiv) and 2- (3- (2-fluoroethoxy)phenyl) acetamide (87.9 mg, 0.446 mmol, 1.10 equiv) in THF (10 mL, 0.041 M) was slowly added (dropwise) t-BuOK solution (114 mg, 1.01 mmol in 10 mL THF, 2.50 equiv) at 0 ° C for 30 min. The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with ice water (5.00 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 ^χ 20.0 mL) . The organic extracts were combined, washed with saturated NaCl (aq) (5.00 mL) , and then dried over Na,?S0₄. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:2) to afford the title compound as a yellow solid (91.6 mg, 0.207 mmol, 51% yield). ^XH NMR (700 MHz, DMSO) δ 11.11 (s, 1H) , 8.09 (s, 1H) , 7.46 (d, J = 8.6 Hz , 1H), 7.31-7.22 (m, 2H) , 7.07-7.02 (m, 1H) , 6.97 (s, 1H), 6.93 (d, J = 7.5 Hz, 1H) , 6.35 (s, 1H) , 4.68-4.64 (m, 1 H ) , 4.61- 4.57 (m, 1H), 4.13-4.09 (m, 1H) , 4.08-4.05 (m, 1H) , 3.89 ( s , 3H) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ): 172.1, 172.0, 157.8, 136.0, 135.7, 128.9, 124.5, 122.6, 116.2, 112.9, 112.5, 102.7, 82.4, 81.9 (d, J = 166.3 Hz), 67.2 (d, J = 19.3 Hz), 33.2. ¹⁹F NMR (376 MH z , ( CD.-);SO, 25 °C, δ) : -224.3 (s) . HRMS (ESI-TOF) (m/z): calcd for C₂iHi-,BrFN_:.0₃ ( [M + H ⁺ ) , 433.0407, found, 433.0421.

Methyl 2- (5-fluoro-l- (2-fluoroethyl) -lH-indol-3-yl) -2-oxoacetate (11a)

TU 2018/013446

64

To a solution of methyl 2- ( 5-fluoro-lJi-indol-3-yl ) -2-oxoacetate (0.55 g, 2.48 mmol, 1.00 equiv) in anhydrous DMF (10.0 mL, 0.248 M) was added sodium hydride (57-63% suspension in mineral oil, 119 mg, 3.00 mmol, 1.21 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 30 min before l-bromo-2-fluoroethanc (378 mg, 3.00 mmol, 1.21 equiv) was added. The reaction mixture was 3tirrcd overnight at room temperature. The reaction was quenched with ice water (40.0 mL) and the aqueous layer was extracted with EtOAc (4 ^χ 30.0 mL) . The organic extracts were combined, washed with 5% LiCl(aq) (5.00 mL) , and then dried over Na₂SO<! . After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:1) to afford the title compound as a yellow solid (0.47 g, 1.76 mmol, 71% yield) .

NMR Spectroscopy: ¾ NMR (700 MHz, CD₃C1, 25 °C, δ) : 8.44 (d, J = 4.3 Hz, 1H) , 8.08-8.12 (m, 1H) , 7.27-7.32 (m, 1H) , 7.04-7.09 (m, 1H) , 4.79 (td, J = 4.6, 46.8 Hz, 2H) , 4.42-4.51 (m, 2H) , 3.95 (d, J = 2.0 Hz, 3H) . ¹³C NMR (175 MHz, CD3CI, 25 °C, δ): 177.0, 163.0, 160.4 (d, J = 238 Hz), 140.9, 133.2, 128.1 (d, J = 11.0 Hz), 113.5 (d, J = 3.4 Hz), 112.8 (d, J = 26.3 Hz), 111.0 (d, J = 9.8 Hz), 108.5 (dd, J = 4.0, 25.0 Hz), 81.5 (d, J = 172.6 Hz), 53.0, 48.0.

Methyl 2- (5-chloro-l- (2-fluoroethyl) -lH-indol-3-yl) -2-oxoacetate

(lib)

11b ^F

To a solution of 2- (5-chloro-lJi-indol-3-yl) -2-oxoacetate (1.70 g, 7.14 mmol, 1.00 equiv) in anhydrous DMF (30.0 mL, 0.238 M) was added sodium hydride (57-63% suspension in mineral oil, 316 mg, 7.90 mmol, 1.11 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 30 mm before l-bromo-2 -fluoroethane (1.09 g, 8.57 mmol, 1.20 equiv) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with ice water (60.0 mL) and the aqueous layer was extracted with EtOAc (6 * 30.0 mL) . The organic extracts were combined, washed with 5% LiCl(aq) (5.00 mL) , and then dried over Na2S0₄. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:1) to afford the title compound as a white solid (700 mg, 2.47 mmol, 35% yield) , NMR Spectroscopy: ^]H R (500 MHz, CDC1₃, 25 °C, δ) ; 8.47 (s, 1H) , 8.45 (d, J = 1.7 Hz, 1H), 7.28-7.35 (m, 2H) , 4.81 (td, J = 4.8, 46.8 Hz, 2H), 4.49 (td, J = 4.8, 26.2 Hz, 2H) , 3.98 (s, 3H) . »C NMR (125 MHz, CDCI₃, 25 °C, δ) : 177.0, 163.0, 140.6, 135.1,130.0, 128.3, 124.9, 122.6, 113.2, 111.0, 81.4 (d, J = 173.2 Hz), 53.0, 47.9 (d, J = 21.3 Hz) .

Methyl-2- (5-fluoro-l- <2-fluoroethyl) -lff-indol-3-yl) -2-oxoacetate

(11c)

To a solution of methyl 2- ( lif-indol-3-yl ) -2-oxoacetate (983 mg, 4.84 mmol, 1.00 equiv) in anhydrous DMF (20.0 mL, 0.242 M) was added sodium hydride (57-63% suspension in mineral oil, 213 mg, 5.32 mmol, 1.10 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 30 min before l-bromo-2-fluoroetiane (737 mg, 5.81 mmol, 1.20 equiv) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched with ice water (40.0 mL) and the aqueous layer was extracted with EtOAc (6 ^χ 30.0 mL) . The organic extracts were combined, washed with 5% LiCl (aq) (5.00 mL ) , and then dried over Na₂S0₄. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel ( EtOAc : hexanes = 1:1) to afford the title compound as a white solid (780 mg, 3.13 mmol, 65% yreld) . NMR Spectroscopy: ^JH NMR (700 MHz, CDC1;,, 25 °C, δ): 8.46-8.50 (m, 1H), 8.44 (s, 1H), 7.32- 7.42 (m, 3H) , 4.80 (td, J = 4.8, 46.8 Hz, 2H), 4.49 (td, J = 4.8, 26.0 Hz, 2H) , 3.97 (s, 3H) . ¹ C NMR (175 MHz, CDCla, 25 °C, δ) : 177.3, 163.3, 140.1, 136.8, 127.3, 124.6, 123.9, 123.2, 113.7, 110.0, 81.5 (J = 172 Hz), 53.0, 47.7 (J = 21.0 Hz).

2- (3-Me hoxyphenyl) acetamide (13a)

To a solution of 2- ( 3-methoxyphenyl ) acetic acid (1.00 g, 6.00 mmol, 1.00 equiv) in dry CH₂C1₂ (20.0 rtiL, 0.300 M) was added thionyl chloride

(5 mL, 69.0 mmol, 11.5 equiv) at 0 °C. The reaction mixture was refluxed for 2 h. The solvent was evaporated and the residue was azeotroped with toluene (2 xlO.O mL. THF (30.0 mL) was added to the residue followed by a slow addition of N¾ ·Η₂0 (7.00 mL, 28-30%) under vigorous stirring at 0 °C. After stirring at room temperature for 2 h, the solvent was removed, Water (10.0 mL) was added and the mixture was heated for 10 min while stirring. The suspension was cooled to 0 °C, filtered and the residue was washed with ice water. The product was dried overnight to afford the title compound as a white solid (580 mg, 3.51 mmol, 59% yield). NMR Spectroscopy: *H NMR (700 MHz, (CD₃)₂SO, 25 °C, δ) : 7.46 (s, 1H) , 7.20 (t, J - 7.8 Hz, 1H) , 6.87 (s, 1H) , 6.83

(s, 1H), 6.82 (s, 1H) , 6.79 (dd, J = 1.9, 8.2 Hz, 1H) , 3.72 (s, 3H) , 3.32 (s, 2H) . ¹³C NMR (175 MHz, (CDsk-SO, 25 °C, δ) : 172.1, 159.1, 138.0, 129.1, 121.3, 114.8, 111.7, 54.9, 42.3. All the analytical data were in good agreement with values reported in the literature

(Kaboudin, B. et al. 2009) .

2- (Naphthalen-l-yl) acetamide (13b)

To a solution of 2- ( naphthalen-l-yl ) acetic acid (1.50 g, 8.01 mmol, 1.00 equiv. ) in dry CH₂C1₂ (10.0 mL, 0.801 M) was added thionyl chloride (6.73 mL, 92.6 mmol, 11.5 equiv.) at 0 °C, then the reaction was refluxed for 2 h. The solvent was evaporated and the residue was azeotroped with toluene (2 xlO.O mL) . To the crude acyl chloride was added THF (30.0 mL) and NH₃ ·Η₂0 (30.0 mL) under vigorous stirring at 0 °C. After the reaction mixture was stirred at room temperature for overnight, the solvent was removed. The residue was purified by column chromatography using with EtOAc:hexane (1 :2 (v/v) ) to afford the title compound as a white solid (1.31 g, 7.07 mmol, 88% yield).

NMR Spectroscopy: ^JH NMR (700 MHz, (CD₃)₂SO, 25 °C, δ): 8.08 (d, J = 8.3 Hz, 1H) , 7.92 (d, J= 7.7 Hz, 1H) , 7.81 (d, J= 7.7 Hz, 1H) , 7.56- 7.49 (m, 3H) , 7.45-7.41 (m, 2H) , 6.99 (s, 1H) , 3.86 (s, 2H) . "C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ): 172.1, 133.3, 132.9, 132.0, 128.3, 127.8, 126.9, 125.9, 125.5, 125.5, 124.2. The ^JH NMR data were in good agreement with values reported in the literature (Tu, T. et al . 2012).

2-(lff-Indol-3-yl)acetamide (13c)

To a solution of indole-3-acetic acid (1.00 g, 5.70 mmol, 1.00 equiv.) in THF (15.0 mL, 0.380 M) was added carbonyl diimidazole (1.02 g, 6.28 mmol, 1.10 equiv.). The resulting solution was stirred at room temperature for 1.5 h. Then NH₃ -H₂0 (20.0 mL) was added, and the reaction was stirred at room temperature for 18 h. The solvent was removed under reduced pressure, and the residue was purified by column chromatography using EtOAc:hexane (2:1 (v/v)). The purification gave the title compound as a white solid (0.902 g, 5.12 mmol, 91% yield). NMR Spectroscopy: Ή NMR (500 MHz, (CD₃)_:S0, 25 °C, δ) : 10.84 (s, 1H) , 7.54 (d, J= 7.9 Hz, 1H) , 7.33 (d, J = 8.1 Hz, 1H), 7.26 (s, 1H) , 7.17 (d, J = 2.3 Hz, 1H) , 7.06-7.03 (m, 1H) , 6.97-6.94 (m, 1H) , 6.80 (s, 1H), 3.45 (s, 2H) . ¹³C NMR (125 MHz, (CD₃) SO, 25 °C, δ): 172.8, 136.0, 127.2, 123.7, 120.8, 118.6, 118.2, 111.2, 109.0, 32.4. All the analytical data were in good agreement with values reported in the literature (Mauger, J. et al . 1989) .

2- (Thiophen-2-yl) acetamide (13d)

12d overnight 13d

72%

To a solution of 2-thiopheneacetic acid (500 mg, 3.52 mmol, 1.00 equiv.) in dry CH2CI2 (10.0 raL) was added thionyl chloride (2.94 mL, 40.4 mmol, 11.5 equiv.) at 0 °C, then the reaction was refluxed for 2 h. The solvent was removed under reduced pressure and the residue was azeotroped with toluene (2 ^χ 5.00 mL) . NH3 -¾0 (10.0 mL) was added to the crude acyl chloride residue in THF (10.0 mL) at 0 °C. After the reaction mixture was stirred at room temperature for overnight, the organic solvent was removed under reduced pressure, and the aqueous mixture was extracted with CH₂C1.₂ (20.0 mL ^χ 3), the organic extracts were dried with MgS0 and concentrated to afford the title compound as a white solid (0.356 g, 2.52 mmol, 72% yield) .

NMR Spectroscopy: ^JH NMR (700 MHz, (CD₃)₂SO, 25 °C, δ) : 7.49 (s, 1H) , 7.33-7.27 (m, 1H) , 6.96 (s, 1H) , 6.93 (t, J = 2.2 Hz, 1H) , 6.88 (s, 1H) , 3.58 (s, 2H) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ) : 171.1, 137.7, 126.5, 126.0, 124.7, 36.3. The ^JH NMR data were in good agreement with values reported in the literature (Gupta, M. et al. 2014) .

2- (2 , -Dichlorophenyl) acetamide (13e)

To a solution of 2- (2, 4-dichlorophenyl) acetic acid (3.60 g, 17.6 mmol,

1.00 equiv) in dry CH₂C1₂ (60.0 mL, 0.293 M) was added thionyl chloride (15.0 mL, 207 mmol, 11.8 equiv) at 0 . C. The reaction mixture was refluxed for 2 h. The solvent was evaporated and the residue was azeotroped with toluene (2 ^χ 5.00 mL) . The residue was dissolved in THF (30.0 mL) and the resulting solution was cooled to 0 °C. NH₃ Ή₂Ο {30.0 mL, 28-30%) was added 3lowly under vigorous stirring. The mixture was warmed Lu room temperature and 3tirrcd for 2 h. Tho solvent was removed, water (10.0 mL) was added and the resulting mixture was heated for 10 min while stirring. The suspension was cooled to 0 °C and filtered. The residue was washed with ice water (20.0 mL) . The product was dried under vacuum for overnight to afford the title compound as a white solid (3.00 g, 14.7 mmol, 84% yield).

NMR Spectroscopy: ¾ NMR (700 MHz , (CD₃)₂SO, 25 °C, δ) : 7.55 (s, 1H) , 7.50 (s, 1H) , 7.30- 7.40 (m, 2H) , 7.02 (s, 1H) , 3.56 (s, 2H) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ): 170.6, 134.6, 133.6, 133.4, 131.9, 128.3, 127.1. The ^JH NMR data was in good agreement with values reported in the literature (M. Wang et al . 2011).

2-Phenylacetamide (13f)

1.2f 89% 13f

To a solution of 2-phenylacetic acid (3.00 g, 22.0 mmol, 1.00 equiv) in dry CH₂C1₂ (70.0 mL, 0.314 M) was added thionyl chloride (19.2 mL, 264 mmol, 12.0 equiv) at 0 °C. The reaction mixture was refluxed for 2 h. The solvent was evaporated and the residue was azeotroped with toluene (2 5 mL) . The residue was dissolved in THF (30.0 mL) and the resulting solution was cooled to 0 °C. NH₃ -H₂0 (40.0 mL, 28-30%) was added slowly under vigorous stirring. The mixture was warmed to room temperature and stirred for 2 h. The solvent was removed, water (20.0 mL) was added and the resulting mixture was heated for 10 min while stirring. The suspension was cooled to 0 °C and filtered. The residue was washed with ice water (20.0 mL) . The product was dried under vacuum overnight to afford the title compound as a white solid (2.66 g, 19.6 mmol, 89% yield) . NMR Spectroscopy: ^!H NMR (700 MHz, (CD₃)₂SO, 25 °C, δ) : 7.49 (s, IH), 7.24-7.36 (m, 4H), 7.20-7.24 (m, IH) , 6.91 (s, IH) , 3.38 (d, J = 2.4 Hz, IH) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ): 172.3, 136.5, 129.1, 128.2, 126.3, 42.3. All the analytical data were in good agreement with values reported in Lhe literature (Allen, C. et al . 2010).

2- (2-Chlorophenyl) acetamide (13g)

120 89% 13g

To a solution of 2-chlorophenylacetic acid 12g (2.00 g, 11.80 mmol, 1.00 equiv.) in dry CH₂C1₂ (30.0 mL) was added thionyl chloride (9.80 ml, 135.3 mmol, 11.5 equiv.) at 0 °C. The reaction was refluxed for 2 h. The solvent was removed under reduced pressure and the residue was azeotroped with toluene (2 5.00 mL) . To a solution of crude acyl chloride in THF (30.0 mL) was added NH₃ -H₂0 (60.0 mL) , and the reaction was stirred at room temperature for overnight. The solvent was removed under reduced pressure, and the precipitate was filtered, washed with water, and dried under vacuum to afford the title compound as a white solid (1.77 g, 10.5 mmol, 89% yield) . NMR Spectroscopy: ¾ NMR (500 MHz, (CD₃)₂SO, 25 °C, δ) : 7.44 (s, IH) , 7.41-7.38 (m, IH) , 7.35-7.33 (m, IH), 7.28-7.24 (m, 2H) , 6.96 (s, IH) , 3.54 (s, 2H) . ¹³C NMR (125 MHz, (CD₃)₂SO, 25 °C, δ): 170.8, 134.3, 133.5, 132.0, 128.8, 128.2,

126.9. The ^JH NMR data was in good agreement with values reported in the literature (O'Connell, J. et al . 2006). 3- (5-Fluoro-l- (2-fluoroet yl) -lH-indol-3-yl) -4- (3-methoxyphenyl) -1H- pyrrole-2 , 5-dione (14a)

Under N2 atmosphere, to a suspension of methyl 2- ( 5-fluoro-1- ( 2 - fluoroethyl) -lH-indol-3-yl) -2 - oxoacetate (153 mg, 0.57 mmol, 1.00 equiv) and 2- { 3-methoxyphenyl ) acetamide (104 mg, 0.627 mmol, 1.10 equiv ) in THF (10.0 mL, 0.057 ) was slowly added (dropwise) t-BuOK solution (154 mg, 1.37 mmol in 15 mL THF, 2 . 40 equiv) at 0 °C for 30 min . The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with ice water (5.00 mL) at 0 °C. The reaction was quenched with HC1 (IN, 4 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 10.0 mL) . The organic extracts were combined, washed with saturated NaCl (aq) (5.00 mL) , and then dried over Na2S0 . After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc:hexanes = 1:2) to afford the title compound as a red solid (142 mg, 0.371 mmol, 65% yield).

NMR Spectroscopy: ^:H NMR (400 MHz, (CD₃)₂SO, 25 °C, δ ) : 11.11 (s, 1H) , 8.13 (s, 1H), 7.59 (dd, J = 4.5, 9.0 Hz, 1H) , 7.24-7.30 (m, 1H) , 6.90- 7.00 (m, 4H), 5.99 (dd, J= 2.5, 10.6 Hz, 1H) , 4.58-4.86 (m, 4H) , 3.58 (s, 3H) . ¹³C NMR (100 MHz, (CD₃)₂SO, 25 °C, δ): 172.1, 172.0, 158.8, 157.0 (d, J = 232.6 Hz), 135.6, 133.2, 131.7, 131.5, 129.3, 129.2, 125. 1 (d, J = 10.6 Hz), 122.1, 115.2, 114.5, 112.0 (d, J = 9.5 Hz), 110.2 (d, J= 25.9 Hz), 106.5 (d, J= 25.1 Hz), 103.8 (d, J= 4.3 Hz), 82.6 (d, J = 166.9 Hz), 55.0, 46.7 (d, J = 19.4 Hz) . ¹⁹F NMR (376 MHz, (CD₃)₂SO, 25 °C, δ): -124.4 (m) , -221.9 (m) . HRMS (ESI-TOF) (m/z): calcd for C₂iH_;7F;N;0;₅ ( [M + H]⁺), 383.1207, found, 383.1211.

3- (5-Chloro-l- (2-fluoroethyl) -lH-indol-3-yl) -4- (3-methoxyphenyl) -1H- pyrrole-2 , 5-dione (14b)

Under N2 atmosphere, to a suspension of methyl 2- (5-chloro-l- (2- fluoroethyl) -lff-indol-3-yl) -2- oxoacetate (281 mg, 0.99 mmol, 1.00 equiv) and 2- (3-methoxyphenyl) acetamide (196 mg, 1.19 mmol, 1.20 equiv) in THF (10.0 mL, 0.099 M) was slowly added (dropwise) t-BuOK solution (333 mg, 2.97 mmol in 15 mL THF, 3.00 equiv) at 0 °C for 30 min. The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with ice water (5.00 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 ^χ 20 mL) . The organic extracts were combined, washed with saturated NaCl (aq) (5.00 mL) , and then dried over Na?S04. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:2) to afford the title compound as a yellow solid (190 mg, 0.476 mmol, 48% yield). NMR Spectroscopy: ¾ NMR (700 MHz, (CD₃)₂SO, 25 °C, δ) : 11.13 (s, 1H), 8.14 (s, 1H) , 7.60 (d, J = 8.6 Hz, 1H) , 7.28 (t, J = 7.9 Hz, 1H) , 7.14 (d, J= 8.6 Hz, 1H) , 7.00 (d, J = 8.2 Hz, 1H) , 6.96 (d, J= 7.6 Hz, 1H), 6.92 (s, 1H) , 6.25 (d, J= 1.4 Hz, 1H) , 4.77 (td, J= 4.4, 47.3 Hz, 2H), 4.65 (td, J = 4.2, 28.0 Hz, 2H) , 3.58 (s, 3H) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ): 172.1, 171.9, 158.9, 135.3, 135.0, 131.6, 131.5, 129.6, 129.3, 125.6., 124.8, 122.1, 122.0, 121.0, 115.3, 114.6, 112.3, 103.5, 82.7 (d, J = 166.8 Hz), 55.0 (d, J = 20.8 Hz), 46.6 (d, <J = 19.4 Hz). ¹⁹F NMR (376 MHz, (CD₃)₂SO, 25 °C, δ) : - 222.0 (m) . HRMS (ESI-TOF) (m/z): calcd for C₂i¾ 7CIFN₂O₃ ( [M + H]⁺), 399.0912, found, 399.0909.

3- (5-Fluoro-l- (2-fluoroethyl) -lH-indol-3-yl) -4- (naphthalen-l-yl) -1H- pyrrole-2 , 5-dione (14c)

11a 14c

To a solution of compound methyl 2- ( 5-fluoro-l~ (2-fluoroethyl) -1H- indol-3-yl ) -2-oxoacetate (27.7 mg, 0.104 mmol, 1.00 equiv.) 2- (naphthalen-l-yl) acetamide (25.0 mg, 0.135 mmol, 1.30 equiv) in THF (5.00 ml, 0.021 ) was added with t-BuOK solution (32.6 mg, 0.291 mmol, 2.20 equiv.) /THF (3.00 mL) under nitrogen atmosphere at 0 °C. After the reaction mixture was stirred at 0 °C for another 5 h, it was quenched with 1 N HCl(aq) (5 mL) . The excess acid was neutralized by sodium bicarbonate, and the solution was extracted with EtOAc (5.00 mL x 3) . The organic layer was washed with water, brine, and dried over a2S04. The reaction mixture was then concentrated and the residue purified by column chromatography using EtOAc:hexane (1:2 (v/v) ) ) to afford the title compound as a yellow solid (23.3 mg, 0.058 mmol, 56% yield). N R Spectroscopy: ²H NMR (700 MHz, (CD₃)₂SO, 25 °C, δ) : 11.19 (s, 1H), 8.19 (s, 1H), 8.05 (d, J = 8.3 Hz, 1H) , 7.97 (d, J = 8.1 Hz, 1H) , 7.71 (d, J = 8.3 Hz, 1H) , 7.57 (t, J = 7.6 Hz, 1H) , 7.49- 7.43 (m, 3H) , 7.35-7.33 (m, J = 2.7 Hz, 1H), 6.82 (dt, J = 2.5, 13.4 Hz, 1H) , 5.63 (dd, J = 2.5, 10.9 Hz, 1H) , 4.73 (q, J = 4.3 Hz, 1H) , 4.66 (q, J = 4.3 Hz, 1H) , 4.61 (t, J = 4.6 Hz, 1H) , 4.56 (t, J = 4.6 Hz, 1H) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ) : 172.3, 172.1, 135.8), 134.4, 133.0, 132.9, 131.1, 129.3, 128.8, 128.7, 128.6, 128.3, 126.3, 126.1, 125.7, 125.6, 125.2, 111.7 (d, J =9.6 Hz), 110.1 (d, J = 26.5 Hz), 105.7 (d, J =■ 25.2 Hz), 104.7 (d, J = 3.8 Hz), 83.0, 82.0, 46.6 (d, J = 19.6 Hz). ¹ F NMR (37b MHz, (CDn^SO, 25 °C, δ): -124.8 ( in ) , -221.9 (m) . HRMS (ESI-TOF) (m/z): calcd for C24H17N2O2 F2 ( [M + H]-), 383.1196, found, 383.1196. 6

74

3 - ( 5-Fluoro-l- (2-fluoroethyl) -lH-indol-3-yl) -4- (lH-indol-3-yl) -1H- pyrrole-2 , 5-dione (14d)

To a solution of compound methyl 2- ( 5-fluoro-l- ( 2-fluoroethyl ) -1H- indol-3-yl ) -2-oxoacetate (100.0 mg, 0.370 mmol, 1.00 equiv.) and 2- (lff-indol-3-yl) acetamide (78.2 mg, 0.450 mmol, 1.20 equiv.) in THF (5.00 mL, 0.074 M) was added with t-BuOK solution (109 mg, 0.970 mmol, 2.60 equiv. ) /THF (5.00 mL) under nitrogen atmosphere at 0 °C. After the reaction mixture was stirred at 0 °C for another 5 h, it was quenched with 1 N HCl(aq) (10.0 mL) . The excess acid was neutralized by sodium bicarbonate, and the solution was extracted with EtOAc (5 mL x 3) . The organic layer was washed with water, brine, and dried over Na₂S0₄. The reaction mixture was concentrated and the residue was purified by column chromatography using EtOAc : hexane (1:1.5 (v/v) ) to afford the title compound as an orange solid (15.0 mg, 0.038 mmol, 10% yield). NMR Spectroscopy: ¾ N R (700 MHz, (CD₃)?SO, 25 °C, δ) : 11.73 (d, J = 2.2 Hz, 1H) , 11.09 (s, 1H) , 7.86 (s, 1H) , 7.79 (d, J = 2.8 Hz, 1H) , 7.52 (q, J = 4.5 Hz, 1H), 7.39 (d, J = 8.1 Hz, 1H) , 7.00 (dt, J = 1.0, 11.6 Hz, 1H), 6.89 (dt, J = 2.6, 9.0 Hz, 1H) , 6.72 (d, J = 8.1 Hz, 1H), 6.63-6.60 (m, 1H) , 6.49 (dd, J = 2.5, 10.4 Hz, 1H) , 5.75 (s, 1H) , 4.71 (t, J= 4.6 Hz, 1H) , 4.65 (t, J = 4.7 Hz, 1H) , 4.60 (t, J = 4.6 Hz, 1H) , 4.55 (t, J = 4.7 Hz, 1H) . ¹³C NMR (175 MHz, (CD₃);_.SO, 25 °C, δ) : 172.8, 157.5, 156.1, 136.0, 133.6, 132.7,129.3, 128.2, 126.4, 125.2, 121.7, 120.7, 119.3, 111.6, 109.9 (d, J = 25.7 Hz), 106.0 (d, J= 24.3 Hz), 105.5 (d, J = 3.8 Hz), 105.3, 83.2, 82.2, 54.9, 46.5 (d, J = 19.4 Hz). ¹⁹F NMR (376 MHz, (CDs SO, 25 °C, δ): - 125.1 (m) , -221.5 (m) . HRMS (ESI-TOF) (m/z): calcd for C₂:H₁₆N₃0_: Fi- ( [M + H] ), 392.1211, found, 392.1207. 3- (5-Fluoro-l- (2-fluoroethyl) -lH-indol-3-yl) -4- (thiophen-2-yl) -1H- pyrrol -2 , 5-dione (14e)

To a solution of compound methyl 2- (5-fluoro-1- ( 2-fluoroethyl ) -1H- indol-3-yl ) -2-oxoacetate (100 mg, 0.370 mmol, 1.00 equiv.) and 2- (thiophen-2-yl) acetamide (69.3 mg, 0.490 mmol, 1.30 equiv.) in THF (5.00 mL, 0.074 M) was added with t-BuOK solution (116 mg, 1.04 mmol, 2.80 equiv.) /THF (5.00 mL) at 0 °C under nitrogen atmosphere. After the reaction mixture was stirred at 0 °C for another 5 h, it was quenched by 1 N HC1 (10.0 mL) . The excess acid was neutralized by sodium bicarbonate. The solution was extracted by EtOAc (10 mL x 3), washed with brine, dried with anhydrous MgSO^, filtered, and concentrated in vacuo. The residue was purified by column chromatography using EtOAc:hexane (1:2 (v/v) ) to afford the title compound a red solid (31.4 mg, 0.088 mmol, 23% yield). NMR Spectroscopy: ¾ NMR (300 MHz, (CD₃)₂SO, 25 °C, δ): 11.19 (s, 1H) , 8.02 (s, 1H), 7.77 (d, J = 5.04 Hz, lH) , 7.67 (q, J = 4.23 Hz, 1H) , 7.26 (d, 3,72 Hz, 1H), 7.10-7.06 (m, 2H) , 6.47 (dd, J = 2.25, 10.29 Hz, 1H) , 4.88 (t, J = 4.47 Hz, 1H) , 4.72-4.69 (m, 2H) , 4.63 (t, J = 4.56 Hz, 1H) . ¹³C NMR (175 Hz, (CD₃)₂SO, 25 °C, δ): 171.7 (d, J = 11.7 Hz), 156.2, 134.8, 133.2, 130.7 (d, J = 5.5 Hz), 130.2, 128.1, 127.1, 125.7, 125.0 (d, J= 10.1 Hz), 112.2, 110.4, 106.4, 103.5, 83.3, 82.0, 46.7 (d, J = 19.7 Hz), 30.6. ¹⁹F NMR (376 MHz, (CD₃)₂SO, 25 °C, δ): - 124.7 (m) , -221.9 (m) . HRMS (ESI-TOF) (m/z): calcd for C:sHi₃N₂0₂SF₂ ( [M + H]⁺), 359.0666, found, 359.0662.

3- (2 , 4-Dichlorophenyl) -4- (5-fluoro-1- (2-fluoroethyl) -lH-indol-3-yl) - lH-pyrrole-2, 5-dione (14f)

Under 2 atmosphere, to a suspension of methyl 2- (5-fluoro-l- (2- fluoroethyl) -lH-indol-3-yl) -2- oxoacetate (377 mg, 1.41 mmol, 1.00 equiv) and 2- (2, 4-dichlorophenyl) acetamide (345 mg, 1.69 mmol, 1.20 equiv) in THF (15.0 mL) was slowly added (dropwise) t-BuOK solution (475 mg, 4.23 mmol in 20.0 mL THF) at 0 °C for 30 min. The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with ice water (5.00 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 ^χ 20.0 mL) . The organic extracts were combined, washed with saturated NaCl (aq) (5.00 mL) , and then dried over Na₂S0₄. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel ( EtOAc : hexanes = 1:2) to afford the title compound as a yellow solid (315 mg, 0.747 mmol, 53% yield). NMR Spectroscopy: 'H NMR (700 MHz, (CD₃)₂SO, 25 °C, δ): 11.26 (s, 1H) , 8.18 (s, 1H) , 7.76 (d, J= 1.8 Hz, 1H) , 7.60 (q, J= 4.4 Hz, 1H) , 7.54 (dd, J = 1.9, 8.3 Hz, 1H) , 7.47 (d, J = 8.3 Hz, 1H) , 6.98-7.08 (m, 1H) , 6.03 (d, J= 10.6 Hz, 1H) , 4.71 (td, J = 4.5, 47.3 Hz, 2H) , 4.64 (td, J = 4.5, 28.0 Hz, 2H) . ¹³C NMR (175 MHz, (CD₃ SO, 25 °C, δ) : 171.7, 171.1, 157.4 (d, J =232.8 Hz), 136.1, 134.9, 134.7, 134.6, 133.6, 133.3, 129.4, 129.1, 127.4, 126.2, 125.7 (d, J = 10.3 Hz), 112.4 (d, J = 10.0 Hz), 110.6 (d, J = 25.7 Hz), 104.7 (d, J = 3.4 Hz), 82.5 (d, J = 166.8 Hz), 46.8 (d, J = 19.3 Hz). ¹⁹F NMR (376 MHz, (CD₃)₂SO, 25 °C, δ) : -124.3 (m) , -222.2 (m) . HRMS (ESI-TOF) (m/z): calcd for C:.₀Hi₃Cl₂F: 2O_? ( [M + H]⁺), 421.0322, found, 421.0317.

3- (2 , 4-Dichlorophenyl) -4- (1- (2-fluoroethyl) -lH-indol-3-yl) -1H- pyrrole-2 , 5-dione (14g)

Under N₂ atmosphere, to a suspension of methyl 2- (1- (2-fluoroethyl) - lH-indol-3-yl) -2-oxoacetate (274 mg, 1.10 mmol, 1.00 equiv) and 2- (2, -dichlorophenyl) acetamide (269 mg, 1.32 mmol, 1.20 equiv) in THF (15.0 mL) was slowly added (dropwise) t-BuOK solution (370 mg, 3.30 mmol in 20.0 mL THF, 3.00 equiv) at 0 °C for 30 min. The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with ice water (5.00 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 ^χ 20.0 mL) . The organic extracts were combined, washed with saturated NaCl (aq) (5.00 mL) , and then dried over Na₂S0 . After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:2) to afford the title compound as a yellow solid (190 mg, 0.471 mmol, 43% yield) . NMR Spectroscopy: ¾ NMR (700 MHz, (CD_)₂S0, 25 °C, δ) : 11.23 (s, 1H) , 8.13 (s, 1H), 7.73 (d, J = 1.0 Hz, 1H) , 7.57 (d, J = 8.2 Hz, 1H) , 7.48 (d, J = 8.3 Hz, 1H), 7.42 (d, J = 8.3 Hz, 1H) , 7.10- 7.16 (m, 1H) , 6.70- 6.85 (m, 1H) , 6.41 (d, J = 8.1 Hz, 1H) , 4.75 (td, J = 4.6, 47.4 Hz, 2H) , 4.64 (td, J = 4.6, 28.0 Hz, 2H) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ) : 171.8, 171.1, 136.6, 135.2, 134.7, 134.6, 134.4, 133.5, 129.6, 129.1, 127.3, 126.1, 125.2, 122.5, 120.7, 120.0, 111.0, 104.6, 82.5 (d, J = 166.7 Hz), 46.5 (d, J = 19.8 Hz) . ¹⁹F NMR (376 MHz, (CD₃)?S0, 25 °C, δ) : -222.2 (m) . HRMS (ESI-TOF) (m/z) : calcd for C₂₀H₁₄CI2FN2O2 ( [M + H]⁺), 403.0388, found, 403.0399. 3- (5-Chloro-l- (2-fluoroethyl) -lH-indol-3-yl) - -phenyl-ltf-pyrrole- 2,5-dione (14h)

Under ₂ atmosphere, to a suspension of methyl 2- (5-chloro-l- (2- fluoroethyl ) -lif-indol-3-yl) -2- oxoacetate (318 mg, 1.12 mmol, 1.00 equiv) and 2-phenylacetamide (182 mg, 1.35 mmol, 1.20 equiv) in THF (10 mL, 0.112 M) was slowly added (dropwise) t-BuOK solution (377 mg, 3.36 mmol in 20 mL THF, 3.00 equiv) at 0 °C for 30 min . The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with ice water (5.00 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 * 20.0 mL) . The organic extracts were combined, washed with saturated NaCl (aq) (5.00 mL) , and then dried over Na₂SO,3. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:2) to afford the title compound as a yellow solid (220 mg, 0.600 mmol, 54% yield).

NMR Spectroscopy: *H NMR (700 MHz, (CD₃)₂SO, 25 °C, δ) : 11.14 (s, 1H) , 8.14 (s, 1H), 7.58 (d, J = 8.7 Hz, 1H) , 7.32-7.45 (m, 5H) , 7.12 (dd, J = 1.6, 8.7 Hz, 1H), 6.19 (d, J = 1.6 Hz, 1H), 4.77 (td, J = 4.5, 40.3 Hz, 2H) , 4.64 (td, J = 4.5, 28.1 Hz, 2H) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ) : 172.2, 172.0, 135.3, 135.1, 131.4, 130.4, 129.9, 129.7, 129.0, 128.2, 125.6, 124.8, 122.1, 120.9, 112.3, 103.5, 82.7 (d, J = 166.7 Hz), 46.7 (d, J = 19.7 Hz). ¹⁹F NMR (376 MHz, (CD₃)2SO, 25 °C, δ) : -221.9 (m) . HRMS (ESI-TOF) (m/z): calcd for C₂oH₁₅Cl FN2O2 ( [M + H]⁺), 369.0806, found, 369.0790.

3- (5-Chloro-l- (2-fluoroethyl) -lH-indol-3-yl) -4- (2-chlorophenyl) -1H- pyrrole-2 , 5-dione (14i)

To a solution of compound methyl 2- (5-chloro-l- (2-fluoroethyl) -1H- indol-3-yl ) -2-oxoacetate (200 mg, 0.710 initio1, 1.00 equiv.) and compound 2- (2-chlorophenyl) acetamide (132 mg, 0.780 mmol, 1.10 equiv.) in THF (10 mL, 0.071 M) was added with t-BuOK solution (190 mg, 1.69 mmol, 2.40 equiv. ) /THF (5.00 mL) at 0 °C under nitrogen atmosphere. After the reaction mixture was stirred at 0 °C for another 5 h, it was quenched by IN HC1 (10.0 mL) . The excess acid was neutralized by sodium bicarbonate. The solution was extracted with EtOAc (10 mL x 3), washed with brine, dried with MgS0₄, filtered, and concentrated in vacuo. The residue was purified through column chromatography using EtOAc : hexanes (1 : 1.5 (v/v) ) to afford the title compound as a light yellow solid (64.8 mg, 0.161 mmOl, 23% yield). NM Spectroscopy: ^lH NMR (700 MHz, (CD₃)₂SO, 25 °C, δ): 11.22 (s, 1H), 8.21 (s, 1H) , 7.58 (d, J = 8.7 Hz, 1H), 7.55 (d, J = 7.9 Hz, 1H) , 7.51-7.49 (m, 1H) , 7.41-7.40 (m, 2H) , 7.13 (dd, J = 2.0, 8.7 Hz, 1H) , 6.19 (d, J = 2.0 Hz, 1H) , 4.76 (t, J = 4.6 Hz, 1H) , 4.70 (t, J = 4.6 Hz, 1H) , 4.65 (t, J = 4.6 Hz, 1H), 4.60 (t, J = 4.6 Hz, 1H) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ) : 171.7, 171.2, 135.7, 135.0, 134.2, 133.5, 132.2, 130.8, 129.4, 127.6, 127.0, 126.2, 125.2, 122.2, 119.8, 112.4, 104.4, 83.0, 82.0, 46.6 (d, J -19.9 Hz). ⁹F NMR (376 MHz, (CD₃)₂SO, 25 °C, δ) : - 222.1 (m) . HRMS (ESI-TOF) (m/z): calcd for C_?QHI4 ₂0₂FC1² ( [M + H]⁺), 403.0416, found, 403.0408. 2- (3- (2- ( (Tetrahydro-2H-pyran-2-yl) oxy) ethoxy) henyl) acetamide

To a solution of 2- (3-hydroxyphenyl) (700 mg, 4.64 mmol, 1.00 equiv) in DMF (10 mL, 0.464 M) was added K₂C0₃ (3.20 g, 23.2 mmol, 5.00 equiv), Nal (139 mg, 0.93 mmol, 0.200 equiv), and 2- (2- bromoethoxy) tetrahydro-2ff-pyranl4 (1.16 g, 5.56 mmol, 1.20 equiv) under nitrogen atmosphere at room temperature. The reaction mixture was heated to 60 °C and stirred for 20 h. The reaction was cooled to 0 °C and quenched with water (50.0 mL) and the aqueous layer was extracted with EtOAc (4 ^χ 30.0 mL) . The organic extracts were combined, washed with 5% LiCl(aq) (5.00 mL) , and then dried over Na2S04. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc) to afford the title compound as a colorless oil (1.05 g, 3.76 mmol, 81% yield). NMR Spectroscopy: ^XH NMR (700 MHz, (CD₃)₂SO, 25 °C, δ): 7.45 (s, 1H) , 7.12-7.23 (m, 1H) , 6.72-6.93 (m, 4H) , 4.65 (d, J = 3.2 Hz, 1H) , 4.06-4.15 (m, 2H) , 3.87- 3.93 (m, 1H), 3.75-3.82 (m, 1H) , 3.67-3.73 (m, 1H) , 3.40-3.47 (m, 1H) , 3.32 (s, 2H) , 1.71 (d, J = 6.4 Hz, 1H) , 1.59-1.65 (m, 1H) , 1.38-1.51 (m, 4H) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ): 172.1, 158.3, 138.0, 129.1, 121.4, 115.5, 112.2, 98.1, 67.0, 65.3, 61.3, 42.3, 30.2, 25.0, 19.1.

3- (l-Ethyl-5-fl oro-lH-indol-3-yl) -4- (3- (2-hydroxyethoxy) henyl) -1H- pyrrole-2 , 5-dione

Under 2 atmosphere, to a suspension of methyl 2- (l-ethyl-5-fluoro- ltf-indol-3-yl) -2-oxoacetate (1.00 g, 4.00 mmol, 1.00 equiv) and 2-(3- (2- ( ( tetrahydro-2ff-pyran-2- yl ) oxy) ethoxy) phenyl ) acetamide (1.12 g, 4.00 mmol, 1.00 equiv) in THF (16 mL, 0.250 equiv) was slowly added (dropwise) t-BuOK solution (900 mg in 70 ml THF, 8.0? mmol, 2.00 equiv) at 0 °C for 30 min. The reaction mixture was stirred at 0 °C for another 4 h. The reaction was quenched with HC1 (IN, 10 mL) at 0 °C and the aqueous layer was extracted with EtOAc (4 ^χ 20 mL) . The combined organic phase was concentrated in vacuo to get crude THP- protected coupling product, which was dissolved in EtOAc:MeOH (v/v = 1:1, 30 mL) and was added HC1 (37%, 3 mL) . The reaction mixture was stirred at room temperature for 3h. The solvent was evaporated in vacuo and the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 2:1) to afford the title compound as a red solid (918 mg, 2.33 mmol, 58% yield).

NMR Spectroscopy: ¾ NMR (700 MHz, (CD₃)₂SO, 25 °C, δ) : 11.09 (s, 1H) , 8.13 (s, 1H) , 7.52 (dd, J = 4.6, 8.9 Hz, 1H) , 7.23 (t, J = 7.9 Hz, 1H) , 7.00 (s, 1H) , 6.93-6.99 (m, 2H) , 6.90 (d, J = 7.6 Hz, 1H) , 5.99 (dd, J = 2.4, 10.5 Hz, 1H) , 4.86 (s, 1H) , 4.30 (q, J = 7.2 Hz, 2H) , 3.84 (t, J = 5.0 Hz, 2H) , 3.63 (d, J = 4.9 Hz, 2H) , 1.39 (t, J = 7.2 Hz, 3H) . ¹³C NMR (175 MHz, (CD₃)₂SO, 25 °C, δ): 172.3, 172.1, 158.3,

157.0 (d, J = 232.5 Hz), 134.8, 132.7, 131.9, 131.7, 129.3, 128.6, 125.2 (d, J = 10.9 Hz), 122.1, 116.0, 115.0, 111.7 (d, J = 9.5 Hz),

110.1 (d, J = 25.8 Hz), 106.6 (d, J = 25.1 Hz), 103.4 (d, J= 4.5 Hz), 69.6, 59.5, 41.3, 15.3. ¹⁹F NMR (376 MHz, (CD₃)₂SO, 25 °C, δ): -124.3

(m) . HRMS (ESI-TOF) (m/z): calcd for C22H20 FN2O4 ( [M + H] ⁽), 395.1407, found, 395.1416.

2- (3- (4- (l-Ethyl-5-fluoro-lH-indol-3-yl) -2 , 5-dioxo-2 , 5-dihydro-lH- pyrrol-3- yl) henoxy) ethyl 4-methylbenzenesulfonate (15)

Under 2 atmosphere, to a suspension of 3- (l-ethyl-5-fluoro-lH-indol- 3-yl) -4- (3- (2-hydroxyethoxy) phenyl) -lii-pyrrole-2 , 5-dione (750 mg, 1.90 mmol, 1.00 equiv) in pyridine (5.00 mL, 0.380 M) was slowly added (dropwise) 4-methylbenzenesulfonyl chloride (TsCl) solution (1.09 g, 5.70 mmol in 5 mL pyridine, 3.00 equiv) at 0 °C for 30 min. After the reaction mixture was stirred at 0 °C for 2 h, to the reaction was added TsCl solution (726 mg, 3.80 mmol in 3.00 mL pyridine, 2.00 equiv) at 0 °C for 15 min. After the reaction mixture was stirred at 0 °C for another 2 h, the reaction was quenched with water (10.0 mL) and the aqueous layer was extracted with EtOAc (3 * 20.0 mL) . The organic extracts were combined, washed with saturated NaCl (aq) (5.00 mL) , and then dried over Na?S0₄. After being concentrated in vacuo, the residue was purified by flash chromatography on silica gel (EtOAc : hexanes = 1:2) to afford the title compound as a yellow solid (570 mg, 1.04 mmol, 55% yield) .

NMR Spectroscopy: ^ΧΗ NMR (700 MHz, (CD₃)₂SO, 25 °C, δ): 11.11 (s, 1H) , 8.14 (s, 1H) , 7.75 (d, J = 8.3 Hz, 2H) , 7.56 (dd, J = 4.6, 9.0 Hz, 1H) , 7.43 (d, J = 8.1 Hz, 2H), 7.22 (t, J = 7.9 Hz, 1H) , 6.86-7.00 (m, 4H), 5.94 (dt, J — 2.5, 10.5 Hz, 1H) , 4.32 (q, J = 7.2 Hz, 2H) , 4.23-4.27 (m, 2H) , 4.00-4.03 (m, 2H) , 2.39 (s, 3H) , 1.40 (t, J = 7.2 Hz, 3H) . ¹³C NMR (175 MHz, (CDs SO, 25 °C, δ): 172.2, 172.0, 157.4, 156.9 (d, J = 232.8 Hz), 145.0, 134.8, 132.7, 132.2, 132.0, 131.7, 130.1, 129.3, 128.4, 127.7, 125.1 (d, J = 10.9 Hz), 122.6, 116.1, 114.8, 111.8 (d, J = 10.0 Hz), 110.1 (d, J = 25.7 Hz), 106.5 (d, J = 25.2 Hz), 103.3 (d, J = 4.5 Hz), 68.9, 65.5, 41.2, 21.1, 15.3. ¹⁹F NMR (376 MHz, (CD₃)₂SO, 25 °C, δ): -224.48 (s). HRMS (ESI-TOF) (m/z): calcd for C:;,H₂₆FN0₆S ( [M + H]'), 549.1475, found, 549.1481. Radiosynthesis of [¹⁸F]10a

Fully automated radiosynthesis of [ ^{1 8} F]10a was performed on a GE Tracerlab FX _FN synthesis module. [ ^{1 8} F ] Fluoride was loaded onto a Waters QMA cartridge and eluted with a solution of K222 (10 mg) and K₂CO3 (1.2 mg) in CIIjC /H?0 (4/1 v/v, 1 mL) into the reactor. The reactor contents are dried under vacuum with holium flow at 95 oC for 4 min, before the addition of CH3CN (1 mL) and the drying continued for 3 min, followed by vacuum alone. The dried [ ^{1 8} F] K/ K222 complex was reacted with 19, at 90 °C, for 10 minutes, to afford the desired [ ^{1 8} F]10a. The tracer, [¹⁸F]10a, was isolated from the automated preparative HPLC (Fig. 1) in 36% radiochemical yield (non-decay corrected) and formulated for injection. The specific activity was >4 Ci/μπιοΐ, and radiochemical purity was >99% at the end of synthesis. In vitro evaluation of GSK-3P inhibition by maleimides .

General Procedure: Commercially available human GSK-3p (BPS Biosciences) was assayed for its ability to phosphorylate the primed peptide substrate (GSP-2: Tyr-Arg-Arg-Ala-Ala-Val-Pro-Pro-Ser-Pro- Ser-Leu-Ser-Arg-His-Ser-Ser-Pro-His-Gln-Ser (PO3H₂) -Glu-AspGlu-Glu- Glu) . ADP-Glo Kinase assay reagents were purchased from Promega (Cat V9102). Kinase reactions were performed with 9 μΜ GSP-2), 15 μΜ ATP, 10 nM GSK3 in reaction buffer (50 m Tris pH 7.5, 5 mM MgCl₂, 0.01% Brij-35, 3 mM DTT ) in the presence of 0-30 μΜ of the maleimides. Test compounds were preincubated with enzyme and substrate for 15 minutes prior to the addition of ATP. Kinase reactions were carried out for a duration of 30 minutes and terminated by addition of ADP-Glo reagent. After 40 minutes incubation with the ADP-Glo reagent, kinase detection reagent was added as per manufacturer's recommendations (Promega). Kinase activity was measured as luminescence resulting due to ADP formation via a multilabel plate reader (Envision, Perkin Elmer) .

In Vivo PET-CT Imaging with [¹⁸F] 10a in Rodents.

All animal studies were performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and were approved by the Massachusetts General Hospital Institutional Animal Care and Use Committee. PET imaging was conducted in Sprague-Dawley rats at baseline (n = 2) and after pretreatment with non-radioactive 10a (1 mg/kg, iv, n = 2) to determine brain uptake and binding specificity. Animals were anesthetized using isoflurane and positioned in the scanner with their heads in the field of view. PET acquisition was initiated at radioLracei administration for GO minutee, followed by CT. Radiotracer doses were 1.06 and 0.62 μθί and 0.92 and 0.67 μθί for animals at baseline and pretreatment conditions, respectively. Dynamic PET data were binned into 33 timeframes and reconstruction of each frame was performed via an iterative MLEM (maximum likelihood expectation maximization) algorithm. Whole brain and regional uptake were quantified using overlaid volumes-of-interest to derive time-activity curves . In Vitro Metabolic Stability Evaluation of 3- (l-ethyl-5-fluoro-lff- indol-3-yl) -4- (3- (2-fluoroethoxy) phenyl) -Iff- pyrrole-2 , 5-dione (10a) in Human and Rat Liver Microsomes .

Materials

Pooled human liver microsomes (pool of 50 donors; Cat # 452165) and pooled male rat liver microsomes (cattt 452501) were purchased from BD Gentest, Woburn, MA. Terfenadine (Cat # T9652, 97.5% pure), quinidine (Q3625), NADPH (Cat # N1630), potassium phosphate monobasic (Cat # P5655, >99-% pure), potassium phosphate dibasic (Cat # P2222, 99% pure) and DMSO (Cat # D5879, >99.5% pure) were purchased from Sigma, Germany. Albendazole (Cat # A4673) and Glipizide (Cat # 50402G) were purchased from Apin Chemicals, Abingdon, UK. 96 well-plates were procured from Axygen, Union City, California. Equipment

Refrigerated centrifuge (Kubota, Tokyo, Japan), water bath (Bio- Techniques, India) and LC-MS/MS Waters Acquity ultra performance LC, API-4000 MDS Sciex, Applied Biosystems, Canada) .

Preparation of reagent (i) Potassium phosphate buffer, 50 mM (pH 7.4) - Potassium phosphate buffer (Kphos) was prepared by adding 0.647 g potassium phosphate monobasic (KH₂PO₄) and 3.527 g potassium phosphate dibasic (K_?HPOi.) to 400 mL of Milli-Q water. pH of the buffer was adjusted to 7.4 and volume was made up to 500 mL. (ii) Preparation of microsomes - Microsomes (20 mg/mL) were diluted in K_Phos buffer to prepare a concentration of 0.178 mg/mL. (iii) Test compound - Stock solutions of 3- (l-ethyl-5-fluoro-lii-indol-3-yl) -4- (3- ( 2-fluoroethoxy) phenyl) - 1Ji-pyrrole-2 , 5-dione (10a) was prepared in DMSO at a concentration of 1 mM. (iv) NADPH solution - A stock solution of 3.33 mM NADPH (3.33X) was prepared by -dissolving appropriate amount of NADPH in Kphos buffer .

Assay Conditions

Total Incubation volume: 100 μL; Compound concentration: 1 μΜ Protein: 0.25 mg/mL; Concentration NADPH: 1 mM; Final DMSO contain: 0.1%; Number of replicates: 2; Time points: 0, 5, 15, 30 and 60 minutes. An 1120 pL aliquot of K_Phos buffer (50 mM, pH 7.4) containing liver microsome (0.357 mg/mL) were added to individual 2 mL tubes (final concentration 0.25 mg/mL) . Test and positive control compounds (1 mM DMSO stocks) were directly spiked into respective tubes to prepare a concentration of 1.428 μΜ (final concentration 1 μΜ) . From the above mix, 70 μL was added to individual wells of 96 well reaction plates and pre-incubated in a 37 °C water bath for 5 min. All the reactions were initiated by adding 30 ih of 3.33 mM NADPH (final concentration 1 mM) . Reactions without NADPH and heat inactivated microsomes minus NADPH (0 min and 60 min) were also incubated to rule out non-NADPH metabolism or chemical instability in the incubation buffer. All reactions were terminated using 100 μL of ice-cold acetonitrile containing internal standard (glipizide, 1 μΜ) at 0, 5, 15, 30 and 60 min. The plates were centrifuged at 4000 RPM for 15 min and 100 L aliquots were submitted for analysis by LC-MS/MS.

Bio-Analysis

Samples were monitored for parent compound disappearance in MRM mode using LC- MS/MS. Example 1. Synthesis of Compounds lOa-k

Target compounds (lOa-k) were synthesized according to procedures (Gunosewoyo, H. et al. 2013; Faul, . M. et al. 1998; Wang, M. et al. 2011) depicLed in Scheme 1.

Scheme 1.

(a) (i) (COC1 ) 2, Et?0, 0 °C to rt, 1 h; (ii) NaOMe, MeOH, -78 °C to rt, 2 h; (b) NaH, DMF, R1X, 0 °C to rt, overnight; (c) (l) MeOH,

H2S04, reflux, 5 h; (ii) NH3 (aq) , rt, overnight; (d) K₂CO_;, DMF, Nal, l-bromo-2-fluoroethane, 60 °C, 20 h; (e) t-BuOK, THF, 0 °C, 4 h.

Commercially available indole derivatives 4a-d were first treated with oxalyl chloride and then with sodium methoxide to afford methylindole- 3-glyoxalates 5a-d. N-Alkylation of 5a-d with alkyl halides in the presence of sodium hydride afforded 6a-k. Preparation of the coupling partner, 2- ( 3- (2-fluoroethoxy ) phenyl ) acetamide 9 began with an acid catalyzed esterification of 2- ( 3-hydroxyphenyl ) acetic acid 7 followed by amidation of the resulting ester with aqueous ammonia to give acetamide 8. O-Alkylation of 8 with l-bromo-2-fluoroethane in the presence of potassium carbonate and sodium iodide afforded 9. With 6a-k and 9 in hand, a condensation reaction using potassium tert- butoxide in tetrahydrofuran at 0 °C was used to provide the desired fluorinated maleimides lOa-k in 35-69% yield. Maintaining reaction temperature at. 0 °C and dropwise addition of the potassium tert- butoxide solution (in THF) were critical in minimizing formation of an alkene side product (HF elimination) and obtaining the desired product in good yields.

Example 2. Synthesis of Compounds 14a-i

The synthetic route to inhibitors containing different aromatic and heteroaromatic moieties at the C4 ' position of the maleimide (14a-i) is illustrated in Scheme 2.

Scheme 2.

14a X^■ f , ¾ «^ m^≠

14b X · C!, * 3-m#t oxypheny1

14c X * F. R₂ » ratfilhatatvl-yl

14d X » F ₂ - 1H4niM^yl

12d, 13d ₂ » fljfX » F, % » 2. <iic loR)phenyl

1¾M «^■& ¾.» 2.4-d¾t*)ropheny(

1*tX » ¾ % » phenyl

130_* i*8¾*2 Moiv≠m≠ 1#X* Cf₈ % « 2*«Noraf)fieny I (a) NaH, DMF, l-bromo-2-fluoroethane, 0 °C to rt, overnight; (b) (i) SOCl₂, CH₂CI₂, rt to reflux, 2 h; (ii) NH₃(aq), rt; overnight; (c) t- BuOK, THF, 0 °C, 4 h. N-Fluoroethylindole derivatives lla-c were prepared from 5a-c and 1 bromo-2-fluoroethane in the presence of sodium hydride in DMF, The amide coupling partners 13a-g were synthesized from the corresponding carboxylic acids 12a-g in a two-step procedure: 12a-g were treated with thionyl chloride to produce acyl chloride intermediates, which were subsequently treated with ammonia to afford acetamides 13a-g. Condensation of N-fluoroethylindole derivatives lla-c with substituted acetamides 13a-g afforded 14a-i in 10-65% yield.

Example 3. SAR Study

With fluorine-substituted maleimides in hand, we first investigated the relationship between indole rings with different substitution patterns and the inhibitory activity of the maleimides (lOa-k) , while keeping the fluoroethyl substituent fixed on the aromatic ring (Scheme 3) .

Scheme 3.

As indicated in Table la, most of the compounds tested showed potent GSK-3 inhibitory activity. The maleimide derivative with an ethyl group on the indole nitrogen and a fluorine atom at the C5- position of the indole ring (10a) was found to be the most potent GSK-3 inhibitor among the whole series, with an IC50 value of 1.70 nM, which is of equivalent potency to the known and highly selective GSK-3p inhibitor CHIR-99021 (IC50 = 1.50 nM) . We observed that both shortening (10b, IC50 = 45.6 nM) and lengthening of the alkyl chain (10c, ICso = 63.9 nM; lOd, IC₅o = 144 nM; lOe, IC₅o > 10, 000 nM) diminished the GSK-3p inhibition. Chemical modifications were carried out to decrease the lipophilicity of the maleimides, which is commonly accomplished by introduction of an oxygen- or nitrogencontaining functional group. The aminoethyl (lOf, IC₅o = 169 nM) and 2- morpholinylethyl (lOg, IC₅₀ = 50.0 nM) analogues were 99- and 29-fold less potent than the ethyl analogue 10a, respectively. Substitution of ethyl by a 2-methoxyethyl group at the indole nitrogen (lOh, IC₅0 = 5.60 nM) resulted in an approximately 3-fold decrease in potency. Comparison of the substituents at the 5-position of indole showed that 5-halogen analogues (10b, IC50 = 45.6 nM; lOj, IC50 = 27.0 nM; and 10k, IC₅₀ = 127 nM) were about 1-6-fold higher in potency than the unsubstituted indole analogue 101 (IC50 = 164 nM) .

Table la.

10b F -CH₂CH₂OCH₃ 5,60

10i H -CH₃ 164

10J Ci -CH₃ 27 0

10k Br -CH₃ 127 IC₅₀ values were experimentally determined under literature methods (Wagner, F. F. 2016) . Kinase reactions were performed with 9 μΜ GSP- 2, 15 μΜ ATP, and 10 nM GSK-3p in reaction buffer (50 mM Tris pH 7.5, 5 mM MgC12, 0.01% Brij-35, 3 mM DTT) in the presence of 0-30 μΜ of the maleimides. For comparison purposes, the IC5₀ values of the known GSK-3 inhibitor CHIR-9902145 in the same assay are shown.

In the next set of SAR modifications, the fluoroethyl group was fixed at indole's nitrogen, while the substituent at the C4 ' position of the maleimide scaffold was varied (Table lb) . 3-methoxyphenyl maleimide analogues 14a and 14b were very potent inhibitors, with IC50 values of 14.7 and 8.80 nM, respectively. Replacement of the 3- methoxyphenyl substituent with either a 1-naphthalenyl (14c) or a 1H- indol-3-yl (14d) moiety was found to be deleterious for the inhibitory activity. Substitution of 3-methoxyphenyl with 2-thiophenyl (14e) resulted in an approximately 2-fold decrease in potency. 2,4- Dichlorophenyl analogues (14f and 14g) significantly diminished GSK- 3β inhibition. Replacement of 3-methoxyphenyl with a phenyl substituent (14h, IC₅₀ = 6.90 nM) rendered an inhibitor with the highest potency in the series. An inhibitor with a 2-chlorophenyl group at the C4 ' position (14i, IC50 = 81.0 nM) was almost 10-fold less potent than the 3-methoxyphenyl analogue 14b.

Table lb.

14f F 2,4-dichlofOphenyl 242

14g H 2,4-dichlorophenyl 1 ,880

14b CI phenyl 6.90

14i C! 2-chlorophenyf 80.9 Compiund 14j (IC50 = 0.84 nM) and compound 14k (IC50 = 0.25 nM) were pre are according to the above methods.

Compiund 141-aa (Table 2) were also prepared according to the above methods .

Table 2. Compounds 141-14aa

Example 4. Jn vivo studies

Based on the in vitro pharmacological results, compound 10a (ICso = 1.70 nM) was selected as the potential PET radioligand for in vivo imaging of 03Κ~3β. The required precursor 15 was prepared according to Scheme 4. Amide 8 was treated with 2- ( 2-bromoethoxy) tetrahydro-2JJ- pyran in the presence of potassium carbonate and sodium iodide to afford the alkylated amide product. Condensation between the amide with glyoxalate 6a followed by removal of the tetrahydro-2fi-pyran (THP) group under acidic conditions gave the alcohol in 58% yield. Finally, O-tosylation of the alcohol with tosyl chloride in the presence of pyridine at 0 °C afforded the desired precursor 15 in 55% yield. Then, fully automated radiosynthesis of [¹⁸F] 10a was performed on a GE Tracerlab FXFN synthesis module. Treatment of precursor 15 with [^1SF]K/K222 complex at 90 °C for 10 min afforded the desired

[¹⁸F]10a in 36% radiochemical yield (non-decay-corrected) with excellent radiochemical purity (>99%) and high specific activity (>148 W

94

GBq/ mol) at the end of the synthesis (Scheme 5) . These results demonstrate the feasibility of synthesizing radiotracers using a fully automated module, which thereby facilitated preclinical studies with

[¹⁸F]10a and is readily translatable to structurally related [¹⁸F]- analogs .

Scheme .

(a) 2CO3, DMF, Nal, 60 °C, overnight; (b) 6a, t-BuOK, THF, 0 °C, 4 h; then HC1 (37%), rt, 4 h; (c) TsCl, Pyridine, 0 °C, 5 h. THP = tetrahydro-2H-pyran.

Scheme 5.

(i) [ ^,fiF] KF, K222 complex, MeCN, 90 °C, 10 min. Ts =toluenesul fonyl , RCY = radiochemical yield.

PET imaging was conducted in Sprague-Dawley rats at baseline (n = 2) and after pretreatment with nonradioactive 10a (1 mg/kg, iv, n = 2) to determine brain uptake and binding saturability (Figures 2 & 3) . Radiotracer uptake in whole-brain was reasonable, reached a peak >0.4%ID/cc at 1-2 min postinfection, and cleared slowly over 60 min. Greater uptake (>0.5%ID/cc) was observed at baseline in certain loci outside of the brain, including harderian glands and tissue near the jaws. Pretreatment did not measurably affect whole-brain radiotracer uptake, but did diminish uptake in other VOIs to ~0. %ID/cc. Although

[¹⁸F] 10a did not display saturable binding in vivo in the rodent brain, these efforts reveal the first report with biological evaluation and imaging studies attempting to develop a [¹⁸F] -labeled GSK-Ββ PET tracer for the central nervous system.

Example 5. Detection of GSK-3P In A Subject

An amount of a composition comprising a compound of the present invention or a salt of the compound, and at least one acceptable carrier, is administered to a subject. The compound is effective to inhibit GSK-3p in the subject.

An amount of a composition comprising a compound of the present invention or a salt of the compound, and at least one acceptable carrier, is administered to a subject. The compound is effective to inhibit GSK-3 in the brain of the subject.

An amount of a composition comprising an ¹⁸F labeled compound of the present invention or a salt of the compound, and at least one acceptable carrier, is administered to a subject. At a period of time after administration, the location of the compound is detected to determine the presense of GSK-3 in the subject.

An amount of a composition comprising an ^ieF labeled compound of the present invention or a salt of the compound, and at least one acceptable carrier, is administered to a subject. At a period of time after administration, the location of the compound is detected to determine the presense of GSK-3p in the brain of the subject. DISCUSSION

Despite the recent promising attempts to study GSK-3p in the CNS, further efforts to quantify 6ΞΚ-3β activity in vivo using brain- penetrating radiotracers are needed. A major drawback of the current PET radiotracers is the use of a short-lived carbon-11 (ti_/z - 20.4 min) , which limits their applications to PET imaging centers equipped with an in-house cyclotron. In contrast, the longer half-life of fluorine-18 (ti_/2 = 109.7 min) compared with that of carbon-11 offers the potential advantage of facilitating widespread use and distribution of the tracer as well as longer imaging protocols to be carried out. In addition, ¹⁸F radioisotope emits positrons with a short positron range, which offers a better spatial resolution and image quality. Thus, development of [ ¹⁸F] -labeled brain-penetrating GSK-3 PET tracers is highly desired (Mossine, A. et al . 2015; Mossine, A. et al. 2016; Pandey, M.K. et al . 2016) .

The maleimide scaffold is highly modular and allows fast assembly of a wide range of molecules through condensation of methylindole-3- glyoxalates (6a-k, lla-c) and (hetero) arylacetamide fragments (9, 13a- g) in a few synthetic steps. Efforts were focused on the synthesis of maleimide derivatives with different aryl/heteroaryl substituents at the C3 ' and C ' positions and compounds with a fluorine atom at a position to which [¹⁸F] fluoride could be incorporated in the PET radiotracer

Embodiments of the invention disclosed herein provide a series of fluorine-substituted maleimide derivatives that are high-affinity GSK-3 inhibitors. The maleimide derivative with an ethyl group on the indole nitrogen and a fluorine atom at the C5-position of the indole ring (e.g. compound 10a) was found to be a potent GSK-3p inhibitor, with an IC₅.1 value of 1.70 nM .

Also disclosed is a radiosynthesis method of a GSR-3p tracer and in vivo PET imaging studies in rodents, which demonstrated that the inhibitors show brain uptake. These results demonstrate that these fluorine-containing maleimides provide a molecular scaffold for potent [¹⁸F] -labeled GSK-3P PET tracers for the study and treatment of GSK- 3β diseases of the central nervous system.

Also disclosed is radiosynthesis melhud of a GSK-3P Lracer [¹⁸F] . Compound [¹⁸F] 10a was synthesized in 36% radiochemical yield (non- decay corrected) with radiochemical purity of >99% and high specific activity of > 148 GBq/μιηοΙ at the end of synthesis. These results demonstrate that in the synthesis of F-18, labelling of structurally related analogs of 10a can be obtained in high yield and high specific activity. In addition, in vivo PET imaging studies in rodents demonstrated that [¹⁸F]10a penetrates the blood-brain barrier. These results indicate that the disclosed fluorine-containing maleimides provide a molecular scaffold for the development of potent [18F]- labeled GSK-3p PET tracers.

The ¹⁸F-labeled PET tracers dislosed herein offer numerous benefits when compared to the existing PET tracers. First, the longer half- life of ¹⁸F (ti/2 = 109.7 min) compared with that of carbon-11 offers an advantage of facilitating widespread use and distribution of the tracer as well as longer imaging protocols. In addition, ¹⁸F radioisotope emits positrons with a short positron range, which offers a better spatial resolution and image quality. The disclsoed PET tracer compositions disclosed herein pass through the blood brain barrier and make use of the long-lived ¹⁸F isotope. Additionally, the disclsoed PET tracer compositions are high-affinity GSK-3 inhibitors and thus are used in the treatment of neurological disorders.

In conclusion, a series of maleimide-based GSK-3 inhibitors containing a fluoroethyl group were prepared for use in in vivo PET imaging. Through structural modifications of the maleimide scaffol at the C3' and C4 ' positions, highly potent GSK-3 inhibitors with nanomolar IC : values were identified. Radiosynthesis of ti^acer [^lsF]10a on an automated module was achieved with excellent radiochemical purity and high specific activity. Thus, using conventional chemical transformations to quickly and reliably access the [¹⁸F] -labeled maleimide-based radiotracers is one of the advantages of the reported chemistry. In vivo PET-CT imaging studies in rodents showed tha [¹⁸F] 10a enters the rodent brain.

REFERENCES

Avila, J. et al . Role of glycogen synthase kinase-3 in Alzheimer's disease pathogenesis and glycogen synthase kinase-3 inhibitors. Expert Rev. Neurother. 2010, 10, 703-10.

Anzini, M. et al . J. Med. Chem. 2013, 56, 3191-3206.

Cohen, P.; Frame, S. The renaissance of GSK3. Nat. Rev. Mol. Cell Biol. 2001, 2, 769-76.

Cohen, P.; Goedert, M . GSK3 inhibitors: development and therapeutic potential. Nat. Rev. Drug Discovery 2004, 3, 479-87.

Copeland, R. A. , Evaluation of enzyme inhibitors in drug discovery: a guide for medicinal chemists and pharmacologists. John Wiley & Sons: 2013, page 131.

Dorn, G. W.; Force, T. Protein kinase cascades in the regulation of cardiac hypertrophy. J. Clin. Invest. 2005, 115, 527-537.

Eldar-Finkelman, H. Glycogen synthase kinase 3: an emerging therapeutic target. Trends Mol. Med. 2002, 8, 126-32.

Eldar-Finkelman, H. et al . Increased glycogen synthase kinase-3 activity in diabetes- and obesity-prone C57BL/6J mice. Diabetes 1999, 48, 1662-6

Faul, M. M. et al . A New, Efficient Method for the Synthesis of Bisindolylmaleimides . J. Org. Chem. 1998, 63, 6053-6058.

Fulmer, G. R. et al . Organometallics . 2010, 29, 2176-2179.

Gunosewoyo, H. et al. Characterization of maleimide-based glycogen synthase kinase-3 (GSK-3) inhibitors as stimulators of steroidogenesis. J. Med. Chem. 2013, 56, 5115-29. Gupta, M.K. et al . , Org. Lett., 2014, 16, 1602.

Haq, S. et al. Glycogen synthase kinase-3 beta is a negative regulator of cardiomyocyte hypertrophy. J. Cell Biol. 2000, 151, 117-129.

Hooper, C. et al. The GSK3 hypothesis of Alzheimer's disease. J. Neuroche . 2008, 104, 1433-9.

Ilovich, 0. et al . Bioorg. Med. Chem., 2010, 18(2), 612.

Jope, R. S . ; Johnson, G. V. The glamour and gloom of glycogen synthase kinase-3. Trends Biochem. Sci. 2004, 29, 95-102.

Kaboudin, B. and Abedi, Y. Synthesis, 2009, 12, 2025.

Kaidanovich-Beilin, 0. Neurological functions of the masterswitch protein kinase-GSK-3 ; Frontiers E-books: 2012.

Kaidanovich-Beilin, 0.; Woodgett, J. R. GSK-3: Functional Insights from Cell Biology and Animal Models. Front. Mol . Neurosci . 2011, 4, 40.

Li, L. et al . Synthesis and Initial in Vivo Studies with [(11)C]SB- 216763: The First Radiolabeled Brain Penetrative Inhibitor of GSK-3. ACS Med. Chem. Lett. 2015, 6, 548-52.

Liana Allen, C. et al . Tetrahedron Lett. 2010, 51(20), 2724.

Liang, S. H. et al . Discovery of a Highly Selective Glycogen Synthase Kinase-3 Inhibitor ( PF-04802367 ) That Modulates Tau Phosphorylation in the Brain: Translation for PET Neuroimaging . Angew. Chem., Int. Ed. 2016, 55, 9601-05.

Luo, J. Glycogen synthase kinase 3beta (GSK3beta) in tumorigenesis and cancer chemotherapy. Cancer Lett. 2009, 273, 194-200. Maixner, D. W. ; Weng, H. R. The Role of Glycogen Synthase Kinase 3 Beta in Neuroinflammation and Pain. J. Pharm. Pharmacol. 2013, 1, 001.

Martinez, A. et al . Glycogen synthase kinase 3 (GSK-3) inhibitors as new promising drugs for diabetes, neurodegeneration, cancer, and inflammation. Med. Res. Rev. 2002, 22, 373-384.

Martins, D. F. et al. The antinociceptive effects of AR-A014418, a selective inhibitor of glycogen synthase kinase-3 beta, in mice. J. Pain 2011, 12, 315-322.

Mauger, J.e tal . Tetrahedron, 1989, 45, 1347.

Miller, P. W. et al . Synthesis of C-ll, F-18, 0-15, and N-13 Radiolabels for Positron Emission Tomography. Angew. Chem. , Int. Ed. 2008, 47, 8998-9033.

Morales-Garcia, J. A. et al . Glycogen Synthase Kinase-3 Inhibitors as Potent Therapeutic Agents for the Treatment of Parkinson Disease. ACS Chem. Neurosci. 2013, 4, 350-360.

Mossine, A. et al . Synthesis and evaluation of 4-[18F] fluorophenacylbromide (4FPB), a selective and irreversible inhibitor of Glycogen Synthase Kinase 3β (GSK-3p) . J. Nucl . Med. 2016, 57, 1099-1099.

Mossine, A. et al . Glycogen Synthase Kinase 3β Inhibitor Scaffolds for PET Radiotracer Development. J. Nucl. Med. 2015, 56, 1094-1094. O'Connell, J. L. et al . Org. Biomol . Chem., 2006, 4, 2716-2723.

Pan, J. Q. et al . AKT Kinase Activity Is Required for Lithium to

Modulate Mood-Related Behaviors in Mice. Neuropsychopharmacology 2011, 36, 1397-1411. Pandey, M. K. ; DeGrado, T. R. Glycogen Synthase Kinase-3 (GSK-3)- Targeted Therapy and Imaging. Theranostics 2016, 6, 571-93. Still, W. C. et al. J. Org. Chem. 1978, 43, 2925-2927.

Tu, T. et al. Green Chem., 2012, 14, 921. Vdlvezan, A. J.; Klein, P. S. GSK-3 and Wnt signaling in neurogenesis and bipoldi disorder. Front. Mol . Ncurocci. 2012 Jan 30;5;1.

Wagner, F. F. et al . Inhibitors of Glycogen Synthase Kinase 3 with Exquisite Kinome-wide Selectivity and their Functional Effects. ACS Chem. Biol. 2016, 11, 1952-63.

Wang, X. et al . J. Am. Chem. Soc. 2013, 135, 10330-10333.

Wang, M. et al . The first synthesis of [ (11) C] SB-216763, a new potential PET agent for imaging of glycogen synthase kinase-3 (GSK- 3). Bioorg. Med. Chem. Lett. 2011, 21, 245-9.

Ye, Q. et al. Bioorg. Med. Chem., 2015, 23, 1179-1188.

Claims

WHAT IS CLAIMED IS:

1. A compound having the structure:

wherein

Ri is -H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl-NH2, alkyl-0 (alkyl) , alkyl-0 ( aryl ) , alkyl-0 (heteroaryl ) , alkyl-NH (alkyl) , alkyl-M(alkyl)₂, alkyl-NH (aryl) , alkyl-N (aryl ) ₂, alkyl- (aryl) (heteroaryl) , alkyl-NH (heteroaryl ) , alkyl- (heteroaryl) ₂, alkyl- S (alkyl), alkyl-S (aryl) , alkyl-S (heteroaryl ) , alkylhalide, alkylaryl, alkylheteroaryl or alkyl- (heterocycloalkyl) ;

R₂, R3, _/ and R5 are each, independently, -H, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl-NH2, alkyl-0 (alkyl) , alkyl- NH (alkyl), alkyl- (alkyl) ₂, alkylhalide, alkylaryl, alkylheteroaryl, alkyl- (heterocycloalkyl) , polyhaloalkyl , OH, 0 (alkyl), O(aryl), O (heteroaryl) , O-alkylhalide, O-polyhaloalkyl , NH₂, NH (alkyl), NH(aryl), NH (heteroaryl) , N(alkyl)₂, (alkyl ) (aryl) , (alkyl ) (heteroaryl ) , N(aryl)₂, N (aryl) (heteroaryl) , N (heteroaryl ) 2, SH, S(alkyl), S(aryl), S (heteroaryl ) , O-polyhaloalkyl, CF₃ or SF=; and

A is an unsubstituted or substituted aryl or heteroaryl, wherein the compound contains at least one alkylhalide group, or a salt of the compound. 2. The compound of claim 1, wherein

Ri is -H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alKyl-NH₂, alkyl-0 (alkyl) , alkyl- (alkyl) :, alkylhalide, alkylaryl, alkylheteroaryl or alkyl- (heterocycloalkyl) ; ί½, 3, Ri, and R5 are each, independently, H or halogen; and A is a unsubstituted or substituted aryl or heteroaryl, wherein the compound contains at least one alkylhalide group, or a salt of the compound.

3. The compound of claim 1 or 2, wherein the compound contains at least one fluoroalkyl group.

4. The compound of claim 1 or 2, wherein the compound contains at least one -(CH2)nF group, wherein n is 2-10.

5. The compound of any one of claims 1-4, wherein A is an unsubstituted or substituted monoaryl or monoheteroaryl .

6. The compound of claim 5, wherein A is an unsubstituted or substituted phenyl, thiophene, furan, pyrrole, indole, benzofuran, benzothiophene, oxazole, isoxazole, imidazole, pyrazole, thiazole, isothiazole, triazole, pyrimidine, pyridazine, pyrazine, or pyridine.

7. The compound of claim 6, wherein A has the structure:

wherein each of Re, R7, Rs, R9 and Rio is, independently, -H, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl-NH2, alkyl-0 (alkyl) , alkyl-NH (alkyl) , alkyl-N (alkyl) 2, alkylhalide, alkylaryl, alkylheteroaryl , alkyl- (heterocycloalkyl ) , polyhaloalkyl, OH, O(alkyl), O(aryl), 0 (heteroaryl ) , O-alkylhalide, O-polyhaloalkyl , N¾, NH(alkyl), NH(aryl), NH (heteroaryl ) , N (alkyl):-, N (alkyl ) (aryl) , N (alkyl ) (heteroaryl ) , N(aryl)₂, N (aryl) (heteroaryl) , (heteroaryl) 2, SH, S (alkyl), S(aryl), S (heteroaryl) , O-polyhaloalkyl, or SF5.

8. The compound of claim 7, wherein each of R₆, R?, Re, 9 and Rio is, independently, -H, halogen, -OH, -NH2, -CF3, -O(alkyl), -0 (haloalkyl) or -NH (alkyl) .

The compound of claim 8, wherein Λ has the structure

10. The compound of claim 7, wherein one of R₆, R7, Rs, R9 and Rio is - OCH2CH2F.

11. The compound of claim 10, wherein A has the structure:

wherein each of Rn, R12, R13, R14 and R15 is, independently, -H, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl-NH₂, alkyl- O(alkyl), alkyl-NH ( alkyl ) , alkyl- (alkyl ) ₂, alkylhalide, alkylaryl, alkylheteroaryl , alkyl- (heterocycloalkyl ) , polyhaloalkyl , OH, 0 (alkyl), O(aryl), 0 (heteroaryl ) , O-alkylhalide, O-polyhaloalkyl, NH₂, NH (alkyl), NH(aryl), NH (heteroaryl ) , N(alkyl)₂, ( alkyl ) ( aryl ) , N ( alkyl ) (heteroaryl ) , N(aryl)₂, ( aryl ) ( heteroaryl ) , ( heteroaryl ) ₂ , SH, S (alkyl), S(aryl), S (heteroaryl) , O-polyhaloalkyl or SF₅.

13. The compound of claim 6 , wherein A has the structure:

wherein each of R-,₆, Rn, R-, s and RIQ is, independently, -H, halogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl-NH2, alkyl-0 (alkyl ) , alkyl-NH (alkyl) , alkyl- (alkyl) 2, alkylhalide, alkylaryl, alkylheteroaryl, alkyl- (heterocycloalkyl) , polyhaloalkyl , OH, O(alkyl), O(aryl), O (heteroaryl) , O-alkylhalide, O-polyhaloalkyl, NH₂, NH (alkyl), NH(aryl), NH (heteroaryl) , N(alkyl)₂, (alkyl ) (aryl) , (alkyl) (heteroaryl ) , N(aryl)₂, N (aryl) (heteroaryl) , N (heteroaryl) 2, SH, S (alkyl), S(aryl), S (heteroaryl) , O-polyhaloalkyl or SF5.

14. The compound of claim 12 , wherein each of R , R_i2 , R13, R14 and R15 is, independently, -H, halogen, -OH, -NH₂, -CF₃, -O(alkyl), -O (alkylhalide) or -NH(alkyl) .

15 . The compound of claim 13 , wherein each of Rie , R17, Ri8 and R19 is, independently, -H, halogen, -OH, -NH₂, -CF₃, -O(alkyl), -0 (alkylhalide) or -NH (alkyl) .

16 . The compound of claim 14 or 15 , wherein A has the structure:

17 . The compound of any one of claims 1 - 16 , wherein Ri is alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkyl-OH, alkyl-NH₂, alkyl-0 (alkyl ) , alkyl, -

NH (alkyl), alkyl- ( alkyl ) ₂, alkylhalide, alkylaryl, alkylheteroaryl or alkyl- (heterocycloalkyl ) .

18. The compound of claim 17, wherein Ri is -CH₃, -CH2CH2, -CH2CH2CH." ,

-CH2CH2CH2CH3, -CH₂CH₂OH, -CH2CH2OCH3, -CH2CH2N (CH₃) 2 or

19. The compound of claim 17, wherein ¾ is

20. The compound of any one of claims 1-19, wherein R₂, R₃, R4, and R5 are each, independently, is H, F, CI or Br.

21. The compound of any one of claims 1-20, wherein Ri is fluoroalkyl, or the aryl or heteroaryl A is substituted with a fluoroalkyl.

22. The compound of any one of claims 1-20, wherein Ri is -CH₂CH₂F or the aryl or heteroaryl A is substituted with a -CH2CH2F.

23. The compound of anyone of claims 3-22, wherein the F of the fluoroalkyl group is ¹⁸F.

24. The compound of claim 1 having the structure:

or a salt of the compound.

25. The compound of claim 1 having the structure:

or a salt of the compound.

26. The compound of claim 1 having the structure:

a salt of the compound.

The compound of claim 1 having the structure

or a salt of the compound.

28. The compound of any one of claims 24-27, wherein the F of the -CH2CH₂F group is ^1BF.

29. A composition comprising the compound of any one of claims 1-28 or a salt of the compound, and at least one acceptable carrier.

30. A composition comprising two compounds of any one of claims 1-28 or a salt of the compounds, and at least one acceptable carrier,

wherein one compound contains a -CH2CH₂F group wherein the F of the -CH2CH2F group is ¹⁸F.

31. A method of inhibiting Glycogen synthase kinase-3P (GSK-3 β) comprising contacting the Glycogen synthase kinase-3 with the compound of any one of claims 1-28, so as to thereby inhibit the GSK-3p.

32. A method of inhibiting Glycogen synthase kinase-3 β (GSK-3 β) in a subject comprising administering to the subject the compound of any one of claims 1-28, so as to thereby inhibit the 63Κ-3β in the subject.

33. The method of claim 32, wherein the Glycogen synthase kinase-3 (GSK- 3β) is located in the brain of the subject.

34. A method of detecting the presence of Glycogen synthase kinase-3 (GSK- 3β) in a subject which comprises determining if an amount of the compound of claim 23 or 28 is present in the subject at a period of time after administration of the compound or salt thereof to the subject, thereby detecting the presence of the Glycogen synthase kinase-3p (GSK-3p) based on the amount of the compound determined to be present in the subject.

35. A method of detecting the presence of Glycogen synthase kinase-3 (GSK- 3β) in the brain of a subject which comprises determining if an amount of the compound of claim 23 or 28 is present in the brain of the subject at a period of time after administration of the compound or salt thereof to the subject, thereby detecting the presence of the Glycogen synthase kinase-3p (GSK-3P ) based on the amount of the compound determined to be present in the brain of the subject.

36. Ά method of detecting the location of Glycogen synthase kinase-3p (GSK- 3β) in the brain of a subject which comprises determining where an amount of the compound of claim 23 or 28 is present in the subject at a period of time after administration of the compound or salt thereof to the subject, thereby detecting the location of the Glycogen synthase kinase-3 (GSK-3P) based on the location of the compound determined to be present in the subject .

37. The method of any one of claims 34-36, further comprising quantifying the amount of the compound in the subject and comparing the quantity to a predetermined control.

38. The method of any one of claims 34-37, wherein the determining is performed by a Positron Emission Tomography (PET) device.

39. The method of any one of claims 34-38, further comprising determining whether the subject is afflicted with a disease associated with dysregulation, up-regulation or down-regulation of Glycogen synthase kinase-3p (GSK-3 ) based on the amount of the compound in the subject.

40. The method of claim 39, wherein the disease associated with dysregulation, up-regulation or down-regulation of Glycogen synthase kinase-3 (GSK-3 ) is Parkinson's disease, Alzheimer's disease (AD), Huntington's disease (HD) , amyotrophic lateral sclerosis (ALS) , bipolar disorder, schizophrenia or major depression.