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Title:
SPIROCYCLIC HETEROCYCLE COMPOUNDS USEFUL AS HIV INTEGRASE INHIBITORS
Document Type and Number:
WIPO Patent Application WO/2015/089847
Kind Code:
A1
Abstract:
Disclosed are Spirocyclic Heterocycle Compounds of Formula (I) and pharmaceutically acceptable salts thereof, where A, B, X, Y, R1, R2 and R11 are as defined herein. Composition comprising at least one Spirocyclic Heterocycle Compound, and methods of using the Spirocyclic Heterocycle Compounds for treating or preventing HIV infection in a subject are also disclosed.

Inventors:
GRAHAM THOMAS H (US)
LIU WENSHENG (US)
YU TAO (US)
ZHANG YONGLIAN (US)
WADDELL SHERMAN T (US)
WAI JOHN S (US)
COLEMAN PAUL J (US)
SANDERS JOHN M (US)
EMBREY MARK W (US)
WALJI ABBAS (US)
FERGUSON RONALD DALE II (US)
MARCO CHRISTINA NG DI (US)
STEELE THOMAS (US)
HU LIHONG (CN)
PENG XUANJIA (CN)
Application Number:
CN2013/090156
Publication Date:
June 25, 2015
Filing Date:
December 20, 2013
Export Citation:
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Assignee:
MERCK SHARP & DOHME (US)
GRAHAM THOMAS H (US)
LIU WENSHENG (US)
YU TAO (US)
ZHANG YONGLIAN (US)
WADDELL SHERMAN T (US)
WAI JOHN S (US)
COLEMAN PAUL J (US)
SANDERS JOHN M (US)
EMBREY MARK W (US)
WALJI ABBAS (US)
FERGUSON RONALD DALE II (US)
MARCO CHRISTINA NG DI (US)
STEELE THOMAS (US)
HU LIHONG (CN)
PENG XUANJIA (CN)
International Classes:
C07D471/20; A61K31/499; A61K31/53; A61K31/55; A61P31/18; C07D471/22; C07D493/20; C07D495/20
Domestic Patent References:
WO2006116764A12006-11-02
WO2007049675A12007-05-03
Foreign References:
EP2540720A12013-01-02
EP1544199A12005-06-22
US7858788B22010-12-28
Attorney, Agent or Firm:
LIU, SHEN & ASSOCIATES (10th Floor, Building 1 10 Caihefang Road, Haidian District, Beijing 0, 100080, CN)
Download PDF:
Claims:
CLAIMS

1. A compound having the formula:

(I)

or a pharmaceutically acceptable salt or prodrug thereof,

wherein:

A is -NHC(O)- or 5 or 6-membered monocyclic heteroaryl;

B is a 3 to 8-membered heterocycloalkyl, which can be optionally be substituted with one or more groups, each independently selected from R7;

X is C1-C3 alkylene;

Y is -C(R3)2- or -N(R4)-;

R1 is selected from Ci-C6 alkyl, C3-C7 cycloalkyl, -(C1-C4 alkylene)-0-(Ci-C6 alkyl), -(C1-C4 alkylene)-S-(Ci-C6 alkyl), -(C C4 alkylene)-S02-(Ci-C6 alkyl), -(C C4 alkylene)-N-(Ci-C6 alkyl)2, Ci-C6 haloalkyl, Ci-C6 hydroxyalkyl, phenyl, 3 to 8-membered monocyclic heterocycloalkyl and 5 or 6-membered monocyclic heteroaryl;

R2 represents up to 3 optional substitutents, each independently selected from halo, Ci-C6 alkyl, -0-(Ci-C6 alkyl) and Ci-C6 haloalkyl;

each occurrence of R3 is independently selected from H, Ci-C6 alkyl, Ci-C6 haloalkyl, C3-C7 cycloalkyl, -(Ci-C4 alkylene)-S-(Ci-C6 alkyl), -(Ci-C4 alkylene)-S02-(Ci-C6 alkyl), -(C1-C4 alkylene)-N-(Ci-C6 alkyl)2, phenyl, 3 to 8-membered monocyclic

heterocycloalkyl and 5 or 6-membered monocyclic heteroaryl;

R4 is selected from Ci-C6 alkyl, -S02R5, -C(0)R5, -(Ci-C6 alkylene)p-C(0)N(R6)2, -(C2-C4 alkylene)-0-(Ci-C6 alkyl), -(C2-C4 alkylene)-S-(Ci-C6 alkyl), -(C2-C4 alkylene)-S02- (Ci-C6 alkyl), -(C2-C4 alkylene)-N-(Ci-C6 alkyl)2, C3-C7 cycloalkyl, phenyl, 4 to 8-membered monocyclic heterocycloalkyl or 6-membered monocyclic heteroaryl and 8 to 10-membered bicyclic heteroaryl;

each occurrence of R5 is independently selected from Ci-C6 alkyl, Ci-C6 haloalkyl, C3-C7 cycloalkyl, phenyl, 3 to 8-membered monocyclic heterocycloalkyl or 6-membered monocyclic heteroaryl and 8 to 10-membered bicyclic heteroaryl, wherein said C3-C7 cycloalkyl group, said phenyl group, said 5 or 6-membered monocyclic heteroaryl group and said 8 to 10- membered bicyclic heteroaryl group can be optionally substituted with one or more groups, each independently selected from R7;

each occurrence of R6 is independently selected from H, Ci-C6 alkyl, C3-C7 cycloalkyl, -(Ci-C6 alkylene)-N(R8)2, Ci-C6 haloalkyl, -C(0)0(Ci-C6 alkyl), -(Ci-C6 alkylene)p- R9 and -(Ci-C6 alkylene)-0-(Ci-C6 alkyl);

each occurrence of R7 is independently selected from halo, Ci-C6 alkyl, Ci-C6 haloalkyl, 3 to 8-membered monocyclic heterocycloalkyl, 6 to 10-membered bicyclic

heterocycloalkyl, -0-(Ci-C6 alkyl), -0-(C6-Cio aryl), -0-(Ci-C6 alkylene)-0-(Ci-C6 alkyl), -O- (Ci-C6 haloalkyl), -0-(Ci-C6 alkylene)-0-(Ci-C6 alkylene)-0-(Ci-C6 alkyl), -NH2, -NH(Ci-C6 alkyl), -N(d-C6 alkyl)2, -S(0)2(C C6 alkyl), -NHS(0)2-(C C6 alkyl), -S(0)2NH-(C C6 alkyl), - OC(0)-(Ci-C6 haloalkyl), -(Ci-C6 alkylene)p-C(0)0-(Ci-C6 alkyl), -(Ci-C6 alkylene)p-C(0)-(Ci- C6 alkyl), -(Ci-C6 alkylene)p-C(0)N(R8)2, Ci-C6 hydroxyalkyl, -P(O)(OR10)2, and -CN;

each occurrence of R8 is independently selected from H, Ci-C6 alkyl, C3-C7 cycloalkyl, C C6 haloalkyl, -C(0)0(d-C6 alkyl), -(C C6 alkylene)p-R9 and -(C C6 alkylene)- 0-(Ci-C6 alkyl);

each occurrence of R9 is independently selected from H, Ci-C6 alkyl, C3-C7 cycloalkyl, 5 or 6-membered monocyclic heteroaryl and 3 to 8-membered monocyclic heterocycloalkyl;

each occurrence of R10 is independently selected from H and Ci-C6 alkyl;

R11 is selected from Ci-C6 alkyl, C3-C7 cycloalkyl and -(C1-C4 alkylene)-0-(C1-

C6 alkyl); and

each occurrence of p is independently 0 or 1. 2. The compound of claim 1 or 2, wherein X is -CH2-.

3. The compound of claim 1 or 2, wherein R11 is H or -CH2OCH3.

The compound of claim 1 having the formula

(la)

or a pharmaceutically acceptable salt thereof,

wherein:

A is: -NHC(O)- or:

B is a 5 or 6-membered heterocycloalkyl;

Y is -CH2- or -N(CH3)-;

R1 is selected from Ci-C6 alkyl and -(C2-C4 alkylene)-0-(Ci-C6 alkyl);

R2 represents up to 2 optional substituents, each independently selected from halo.

The compound of any of claims 1-3, wherein, wherein Y is -CH2- or -N(CH3)-.

The compound of any of claims 1-5, wherein A is -NH-C(O)-.

The compound of any of claims 1-5, wherein A is:

8. The compound of any of claims 1-7, wherein B is a 5 or 6-membered monocyclic heterocycloalkyl ring.

9. The compound of claim 8, wherein B is tetrahydrofuranyl or tetrahydropyranyl.

10. The compound of claim 8, wherein B is piperidinyl, optionally substituted on the ring nitrogen atom with -C(0)OR5, -C(0)R5, -S(0)2-(Ci-C6 alkyl) or -S(0)2NH-(Ci-C6 alkyl).

11. The compound of any of claims 1-10, wherein R1 is selected from methyl, ethyl, n-propyl and -CH2CH2OCH3.

12. The compound of any of claims 1-11, wherein R represents two fluoro groups, the ortho and para positions.

13. A compound having the structure:

88

89

or a pharmaceutically acceptable salt thereof.

14. A pharmaceutical composition comprising an effective amount of a compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

15. A method for the inhibition of HIV integrase in a subject in need thereof which comprises administering to the subject an effective amount of the compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof.

16. A method for the treatment of infection by HIV or for the treatment, prophylaxis, or delay in the onset or progression of AIDS in a subject in need thereof, which comprises administering to the subject an effective amount of the compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof.

17. A compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, for use in therapy.

18. A compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, for use in the preparation of a medicament for the inhibition of HIV integrase, for the treatment or prophylaxis of infection by HIV, or for the treatment, prophylaxis, or delay in the onset or progression of AIDS in a subject in need thereof.

19. The pharmaceutical composition of claim 14, further comprising one or more additional therapeutic agents selected from raltegravir, lamivudine, abacavir, ritonavir, dolutegravir, arunavir, atazanavir, emtricitabine, tenofovir, elvitegravir, rilpivirine and lopinavir.

20. The method of claim 16, further comprising administering to the subject one or more additional therapeutic agents selected from raltegravir, abacavir, lamivudine, ritonavir and lopinavir, wherein the amounts administered of the compound of any one of claims 1 to 13 and the one or more additional therapeutic agents, are together effective to treat infection by HIV or to treat, prevent or delay the onset or progression of AIDS.

Description:
SPIROCYCLIC HETEROCYCLE COMPOUNDS USEFUL AS HIV INTEGRASE

INHIBITORS

FIELD OF THE INVENTION

The present invention relates to Spirocyclic Heterocycle Compounds, compositions comprising at least one Spirocyclic Heterocycle Compound, and methods of using the Spirocyclic Heterocycle Compounds for treating or preventing HIV infection in a subject.

BACKGROUND OF THE INVENTION

A retrovirus designated human immunodeficiency virus (HIV), particularly the strains known as HIV type-1 (HIV-1) virus and type-2 (HIV-2) virus, is the etiological agent of the complex disease that includes progressive destruction of the immune system (acquired immune deficiency syndrome; AIDS) and degeneration of the central and peripheral nervous system. A common feature of retrovirus replication is the insertion by virally-encoded integrase of +proviral DNA into the host cell genome, a required step in HIV replication in human T- lymphoid and monocytoid cells. Integration is believed to be mediated by integrase in three steps: assembly of a stable nucleoprotein complex with viral DNA sequences; cleavage of two nucleotides from the 3' termini of the linear proviral DNA; covalent joining of the recessed 3' OH termini of the proviral DNA at a staggered cut made at the host target site. The fourth step in the process, repair synthesis of the resultant gap, may be accomplished by cellular enzymes.

Nucleotide sequencing of HIV shows the presence of a pol gene in one open reading frame [Ratner, L. et al, Nature, 313, 277(1985)]. Amino acid sequence homology provides evidence that the pol sequence encodes reverse transcriptase, integrase and an HIV protease [Tohours, H. et al, EMBO J. 4, 1267 (1985); Power, M.D. et al, Science, 231, 1567 (1986); Pearl, L.H. et al, Nature, 329, 351 (1987)]. All three enzymes have been shown to be essential for the replication of HIV.

It is known that some antiviral compounds which act as inhibitors of HIV replication are effective agents in the treatment of AIDS and similar diseases, including reverse transcriptase inhibitors such as azidothymidine (AZT) and efavirenz and protease inhibitors such as indinavir and nelfmavir. The compounds of this invention are inhibitors of HIV integrase and inhibitors of HIV replication.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides Compounds of Formula (I):

OH O or a pharmaceutically acceptable salt or prodrug thereof,

wherein:

A is -NHC(O)- or 5 or 6-membered monocyclic heteroaryl;

B is a 3 to 8-membered heterocycloalkyl, which can be optionally be substituted with one or more groups, each independently selected from R 7 ;

X is C1-C3 alkylene;

Y is -C(R 3 ) 2 - or -N(R 4 )-;

R 1 is selected from Ci-C 6 alkyl, C3-C7 cycloalkyl, -(C1-C4 alkylene)-0-(Ci-C 6 alkyl), -(C 1 -C4 alkylene)-S-(Ci-C 6 alkyl), -(C C 4 alkylene)-S0 2 -(Ci-C 6 alkyl), -(C C 4 alkylene)-N-(Ci-C6 alkyl) 2 , Ci-C 6 haloalkyl, Ci-C 6 hydroxyalkyl, phenyl, 3 to 8-membered monocyclic heterocycloalkyl and 5 or 6-membered monocyclic heteroaryl;

R 2 represents up to 3 optional substitutents, each independently selected from halo, Ci-C 6 alkyl, -0-(C C 6 alkyl) and C C 6 haloalkyl;

each occurrence of R 3 is independently selected from H, Ci-C 6 alkyl, Ci-C 6 haloalkyl, C3-C7 cycloalkyl, -(Ci-C 4 alkylene)-S-(Ci-C 6 alkyl), -(Ci-C 4 alkylene)-S0 2 -(Ci-C 6 alkyl), -(C1-C4 alkylene)-N-(Ci-C6 alkyl) 2 , phenyl, 3 to 8-membered monocyclic

heterocycloalkyl and 5 or 6-membered monocyclic heteroaryl;

R 4 is selected from Ci-C 6 alkyl, -S0 2 R 5 , -C(0)R 5 , -(Ci-C 6 alkylene) p -C(0)N(R 6 ) 2 ,

-(C2-C4 alkylene)-0-(Ci-C 6 alkyl), -(C 2 -C 4 alkylene)-S-(Ci-C 6 alkyl), -(C 2 -C 4 alkylene)-S0 2 - (Ci-C 6 alkyl), -(C 2 -C 4 alkylene)-N-(Ci-C6 alkyl) 2 , C3-C7 cycloalkyl, phenyl, 4 to 8-membered monocyclic heterocycloalkyl or 6-membered monocyclic heteroaryl and 8 to 10-membered bicyclic heteroaryl;

each occurrence of R 5 is independently selected from Ci-C 6 alkyl, Ci-C 6 haloalkyl,

C3-C 7 cycloalkyl, phenyl, 3 to 8-membered monocyclic heterocycloalkyl or 6-membered monocyclic heteroaryl and 8 to 10-membered bicyclic heteroaryl, wherein said C3-C 7 cycloalkyl group, said phenyl group, said 5 or 6-membered monocyclic heteroaryl group and said 8 to 10- membered bicyclic heteroaryl group can be optionally substituted with one or more groups, each independently selected from R 7 ; each occurrence of R 6 is independently selected from H, Ci-C 6 alkyl, C 3 -C 7 cycloalkyl, -(Ci-C 6 alkylene)-N(R 8 ) 2 , Ci-C 6 haloalkyl, -C(0)0(Ci-C 6 alkyl), -(Ci-C 6 alkylene) p - R 9 and -(C C 6 alkylene)-0-(Ci-C 6 alkyl);

each occurrence of R 7 is independently selected from halo, Ci-C 6 alkyl, Ci-C 6 haloalkyl, 3 to 8-membered monocyclic heterocycloalkyl, 6 to 10-membered bicyclic

heterocycloalkyl, -0-(Ci-C 6 alkyl), -O-(C 6 -Ci 0 aryl), -0-(Ci-C 6 alkylene)-0-(Ci-C 6 alkyl), -O- (Ci-C 6 haloalkyl), -0-(C C 6 alkylene)-0-(Ci-C 6 alkylene)-0-(Ci-C 6 alkyl), -NH 2 , -NH(C C 6 alkyl), -N(Ci-C 6 alkyl) 2 , -S(0) 2 (Ci-C 6 alkyl), -NHS(0) 2 -(Ci-C 6 alkyl), -S(0) 2 NH-(Ci-C 6 alkyl), - OC(0)-(Ci-C 6 haloalkyl), -(Ci-C 6 alkylene) p -C(0)0-(Ci-C 6 alkyl), -(Ci-C 6 alkylene) p -C(0)-(Ci- C 6 alkyl), -(Ci-C 6 alkylene) p -C(0)N(R 8 ) 2 , Ci-C 6 hydroxyalkyl, -P(O)(OR 10 ) 2 , and -CN;

each occurrence of R 8 is independently selected from H, Ci-C 6 alkyl, C 3 -C 7 cycloalkyl, Ci-C 6 haloalkyl, -C(0)0(Ci-C 6 alkyl), -(Ci-C 6 alkylene) p -R 9 and -(Ci-C 6 alkylene)- 0-(Ci-C 6 alkyl);

each occurrence of R 9 is independently selected from H, Ci-C 6 alkyl, C 3 -C 7 cycloalkyl, 5 or 6-membered monocyclic heteroaryl and 3 to 8-membered monocyclic heterocycloalkyl;

each occurrence of R 10 is independently selected from H and Ci-C 6 alkyl;

R 11 is selected from Ci-C 6 alkyl, C3-C7 cycloalkyl and -(C1-C4 alkylene)-0-(C 1 -

C 6 alkyl); and

each occurrence of p is independently 0 or 1.

The Compounds of Formula (I) (also referred to herein as the "Spirocyclic Heterocycle Compounds") and pharmaceutically acceptable salts thereof, can be useful, for example, for inhibiting HIV viral replication or replicon activity, and for treating or preventing HIV infection in a subject. Without being bound by any specific theory, it is believed that the Spirocyclic Heterocycle Compounds inhibit HIV viral replication by inhibiting HIV Integrase.

Accordingly, the present invention provides methods for treating or preventing HIV infection in a subject, comprising administering to the subject an effective amount of at least one Spirocyclic Heterocycle Compound.

The details of the invention are set forth in the accompanying detailed description below.

Although any methods and materials similar to those described herein can be used in the practice or testing of the present invention, illustrative methods and materials are now described. Other embodiments, aspects and features of the present invention are either further described in or will be apparent from the ensuing description, examples and appended claims.

DETAILED DESCRIPTION OF THE INVENTION

The present invention includes to Spirocyclic Heterocycle Compounds, compositions comprising at least one Spirocyclic Heterocycle Compound, and methods of using the Spirocyclic Heterocycle Compounds for treating or preventing HIV infection in a subject.

Definitions and Abbreviations

The terms used herein have their ordinary meaning and the meaning of such terms is independent at each occurrence thereof. That notwithstanding and except where stated otherwise, the following definitions apply throughout the specification and claims. Chemical names, common names, and chemical structures may be used interchangeably to describe the same structure. These definitions apply regardless of whether a term is used by itself or in combination with other terms, unless otherwise indicated. Hence, the definition of "alkyl" applies to "alkyl" as well as the "alkyl" portions of "hydroxyalkyl," "haloalkyl," "-O-alkyl," etc...

As used herein, and throughout this disclosure, the following terms, unless otherwise indicated, shall be understood to have the following meanings:

A "subject" is a human or non-human mammal. In one embodiment, a subject is a human. In another embodiment, a subject is a primate. In another embodiment, a subject is a monkey. In another embodiment, a subject is a chimpanzee. In still another embodiment, a subject is a rhesus monkey.

The term "effective amount" as used herein, refers to an amount of Spirocyclic Heterocycle Compound and/or an additional therapeutic agent, or a composition thereof that is effective in producing the desired therapeutic, ameliorative, inhibitory or preventative effect when administered to a subject suffering from HIV infection or AIDS. In the combination therapies of the present invention, an effective amount can refer to each individual agent or to the combination as a whole, wherein the amounts of all agents administered are together effective, but wherein the component agent of the combination may not be present individually in an effective amount.

The term "preventing," as used herein with respect to an HIV viral infection or AIDS, refers to reducing the likelihood or severity of HIV infection or AIDS.

The term "alkyl," as used herein, refers to an aliphatic hydrocarbon group having one of its hydrogen atoms replaced with a bond. An alkyl group may be straight or branched and contain from about 1 to about 20 carbon atoms. In one embodiment, an alkyl group contains from about 1 to about 12 carbon atoms. In different embodiments, an alkyl group contains from 1 to 6 carbon atoms (Ci-C 6 alkyl) or from about 1 to about 4 carbon atoms (C 1 -C4 alkyl). Non- limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, neopentyl, isopentyl, n-hexyl, isohexyl and neohexyl. An alkyl group may be unsubstituted or substituted by one or more substituents which may be the same or different, each substituent being independently selected from the group consisting of halo, alkenyl, alkynyl, aryl, cycloalkyl, cyano, hydroxy, -O-alkyl, -O-aryl, -alkylene-O-alkyl, alkylthio, -NH 2 , -NH(alkyl), -N(alkyl) 2 , -NH(cycloalkyl), -0-C(0)-alkyl, -0-C(0)-aryl, -O-C(O)- cycloalkyl, -C(0)OH and -C(0)0-alkyl. In one embodiment, an alkyl group is linear. In another embodiment, an alkyl group is branched. Unless otherwise indicated, an alkyl group is unsubstituted.

The term "alkenyl," as used herein, refers to an aliphatic hydrocarbon group containing at least one carbon-carbon double bond and having one of its hydrogen atoms replaced with a bond. An alkenyl group may be straight or branched and contain from about 2 to about 15 carbon atoms. In one embodiment, an alkenyl group contains from about 2 to about 12 carbon atoms. In another embodiment, an alkenyl group contains from about 2 to about 6 carbon atoms. Non-limiting examples of alkenyl groups include ethenyl, propenyl, n-butenyl, 3- methylbut-2-enyl, n-pentenyl, octenyl and decenyl. An alkenyl group may be unsubstituted or substituted by one or more substituents which may be the same or different, each substituent being independently selected from the group consisting of halo, alkenyl, alkynyl, aryl, cycloalkyl, cyano, hydroxy, -O-alkyl, -O-aryl, -alkylene-O-alkyl, alkylthio, -NH 2 , -NH(alkyl), -N(alkyl) 2 , - NH(cycloalkyl), -0-C(0)-alkyl, -0-C(0)-aryl, -0-C(0)-cycloalkyl, -C(0)OH and -C(0)0-alkyl. The term "C 2 -C 6 alkenyl" refers to an alkenyl group having from 2 to 6 carbon atoms. Unless otherwise indicated, an alkenyl group is unsubstituted.

The term "alkynyl," as used herein, refers to an aliphatic hydrocarbon group containing at least one carbon-carbon triple bond and having one of its hydrogen atoms replaced with a bond. An alkynyl group may be straight or branched and contain from about 2 to about 15 carbon atoms. In one embodiment, an alkynyl group contains from about 2 to about 12 carbon atoms. In another embodiment, an alkynyl group contains from about 2 to about 6 carbon atoms. Non-limiting examples of alkynyl groups include ethynyl, propynyl, 2-butynyl and 3- methylbutynyl. An alkynyl group may be unsubstituted or substituted by one or more

substituents which may be the same or different, each substituent being independently selected from the group consisting of halo, alkenyl, alkynyl, aryl, cycloalkyl, cyano, hydroxy, -O-alkyl, - O-aryl, -alkylene-O-alkyl, alkylthio, -NH 2 , -NH(alkyl), -N(alkyl) 2 , -NH(cycloalkyl), -O-C(O)- alkyl, -0-C(0)-aryl, -0-C(0)-cycloalkyl, -C(0)OH and -C(0)0-alkyl. The term "C 2 -C 6 alkynyl" refers to an alkynyl group having from 2 to 6 carbon atoms. Unless otherwise indicated, an alkynyl group is unsubstituted.

The term "alkylene," as used herein, refers to an alkyl group, as defined above, wherein one of the alkyl group's hydrogen atoms has been replaced with a bond. Non- limiting examples of alkylene groups include -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, - CH(CH 3 )CH 2 CH 2 -, -CH(CH 3 )- and -CH 2 CH(CH 3 )CH 2 -. In one embodiment, an alkylene group has from 1 to about 6 carbon atoms. In another embodiment, an alkylene group has from about 3 to about 5 carbon atoms. In another embodiment, an alkylene group is branched. In another embodiment, an alkylene group is linear. In one embodiment, an alkylene group is -CH 2 -. The term "C 1-C4 alkylene" refers to an alkylene group having from 1 to 4 carbon atoms. The term "C 2 -C 4 alkylene" refers to an alkylene group having from 2 to 4 carbon atoms.

The term "alkenylene," as used herein, refers to an alkenyl group, as defined above, wherein one of the alkenyl group's hydrogen atoms has been replaced with a bond. Non- limiting examples of alkenylene groups include -CH=CH-, -CH=CHCH 2 -, -CH 2 CH=CH-, - CH 2 CH=CHCH 2 -, -CH=CHCH 2 CH 2 -, -CH 2 CH 2 CH=CH- and -CH(CH 3 )CH=CH-. In one embodiment, an alkenylene group has from 2 to about 6 carbon atoms. In another embodiment, an alkenylene group has from about 3 to about 5 carbon atoms. In another embodiment, an alkenylene group is branched. In another embodiment, an alkenylene group is linear. The term "C 2 -C 6 alkylene" refers to an alkenylene group having from 2 to 6 carbon atoms. The term "C 3 - C 5 alkenylene" refers to an alkenylene group having from 3 to 5 carbon atoms.

The term "aryl," as used herein, refers to an aromatic monocyclic or multicyclic ring system comprising from about 6 to about 14 carbon atoms. In one embodiment, an aryl group contains from about 6 to about 10 carbon atoms. An aryl group can be optionally substituted with one or more "ring system substituents" which may be the same or different, and are as defined herein below. In one embodiment, an aryl group can be optionally fused to a cycloalkyl or cycloalkanoyl group. Non-limiting examples of aryl groups include phenyl and naphthyl. In one embodiment, an aryl group is phenyl. Unless otherwise indicated, an aryl group is unsubstituted.

The term "arylene," as used herein, refers to a bivalent group derived from an aryl group, as defined above, by removal of a hydrogen atom from a ring carbon of an aryl group. An arylene group can be derived from a monocyclic or multicyclic ring system comprising from about 6 to about 14 carbon atoms. In one embodiment, an arylene group contains from about 6 to about 10 carbon atoms. In another embodiment, an arylene group is a naphthylene group. In another embodiment, an arylene group is a phenylene group. An arylene group can be optionally substituted with one or more "ring system substituents" which may be the same or different, and are as defined herein below. An arylene group is divalent and either available bond on an arylene group can connect to either group flanking the arylene group. For example, the group "A-arylene-B," wherein the arylene group is:

is understood to represent both:

In one embodiment, an arylene group can be optionally fused to a cycloalkyl or cycloalkanoyl group. Non-limiting examples of arylene groups include phenylene and naphthalene. In one embodiment, an arylene group is unsubstituted. In another embodiment, an arylene group is:

Unless otherwise indicated, an arylene group is unsubstituted.

The term "cycloalkyl," as used herein, refers to a non-aromatic mono- or multicyclic ring system comprising from about 3 to about 10 ring carbon atoms. In one embodiment, a cycloalkyl contains from about 5 to about 10 ring carbon atoms. In another embodiment, a cycloalkyl contains from about 3 to about 7 ring atoms. In another embodiment, a cycloalkyl contains from about 5 to about 6 ring atoms. The term "cycloalkyl" also encompasses a cycloalkyl group, as defined above, which is fused to an aryl (e.g., benzene) or heteroaryl ring. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. Non-limiting examples of multicyclic cycloalkyls include 1-decalinyl, norbornyl and adamantyl. A cycloalkyl group can be optionally substituted with one or more "ring system substituents" which may be the same or different, and are as defined herein below. In one embodiment, a cycloalkyl group is unsubstituted. The term "3 to 7-membered cycloalkyl" refers to a cycloalkyl group having from 3 to 7 ring carbon atoms. Unless otherwise indicated, a cycloalkyl group is unsubstituted. A ring carbon atom of a cycloalkyl group may be functionalized as a carbonyl group. An illustrative example of such a cycloalkyl group (also referred to herein as a "cycloalkanoyl" group) includes, but is not limited to, cyclobutanoyl:

The term "halo," as used herein, means -F, -CI, -Br or -I.

The term "haloalkyl," as used herein, refers to an alkyl group as defined above, wherein one or more of the alkyl group's hydrogen atoms has been replaced with a halogen. In one embodiment, a haloalkyl group has from 1 to 6 carbon atoms. In another embodiment, a haloalkyl group is substituted with from 1 to 3 F atoms. Non-limiting examples of haloalkyl groups include -CH 2 F, -CHF 2 , -CF 3 , -CH 2 C1 and -CC1 3 . The term "Ci-C 6 haloalkyl" refers to a haloalkyl group having from 1 to 6 carbon atoms.

The term "hydroxyalkyl," as used herein, refers to an alkyl group as defined above, wherein one or more of the alkyl group's hydrogen atoms have been replaced with an - OH group. In one embodiment, a hydroxyalkyl group has from 1 to 6 carbon atoms. Non- limiting examples of hydroxyalkyl groups include -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH and -CH 2 CH(OH)CH 3 . The term "Ci-C 6 hydroxyalkyl" refers to a hydroxyalkyl group having from 1 to 6 carbon atoms.

The term "heteroaryl," as used herein, refers to an aromatic monocyclic or multicyclic ring system comprising about 5 to about 14 ring atoms, wherein from 1 to 4 of the ring atoms is independently O, N or S and the remaining ring atoms are carbon atoms. In one embodiment, a heteroaryl group has 5 to 10 ring atoms. In another embodiment, a heteroaryl group is monocyclic and has 5 or 6 ring atoms. In another embodiment, a heteroaryl group is bicyclic. A heteroaryl group can be optionally substituted by one or more "ring system substituents" which may be the same or different, and are as defined herein below. A heteroaryl group is joined via a ring carbon atom, and any nitrogen atom of a heteroaryl can be optionally oxidized to the corresponding N-oxide. The term "heteroaryl" also encompasses a heteroaryl group, as defined above, which is fused to a benzene ring. Non-limiting examples of heteroaryls include pyridyl, pyrazinyl, furanyl, thienyl, pyrimidinyl, pyridone (including N-substituted pyridones), isoxazolyl, isothiazolyl, oxazolyl, oxadiazolyl, thiazolyl, pyrazolyl, furazanyl, pyrrolyl, triazolyl, 1 ,2,4-thiadiazolyl, pyrazinyl, pyridazinyl, quinoxalinyl, phthalazinyl, oxindolyl, imidazo[l ,2-a]pyridinyl, imidazo[2,l-b]thiazolyl, benzofurazanyl, indolyl, azaindolyl, benzimidazolyl, benzothienyl, quinolinyl, imidazolyl, benzimidazolyl, thienopyridyl, quinazolinyl, thienopyrimidyl, pyrrolopyridyl, imidazopyridyl, isoquinolinyl, benzoazaindolyl, 1 ,2,4-triazinyl, benzothiazolyl and the like, and all isomeric forms thereof. The term

"heteroaryl" also refers to partially saturated heteroaryl moieties such as, for example, tetrahydroisoquinolyl, tetrahydroquinolyl and the like. In one embodiment, a heteroaryl group is a 5-membered heteroaryl. In another embodiment, a heteroaryl group is a 6-membered monocyclic heteroaryl. In another embodiment, a heteroaryl group comprises a 5- to 6- membered monocyclic heteroaryl group fused to a benzene ring. Unless otherwise indicated, a heteroaryl group is unsubstituted.

The term "heterocycloalkyl," as used herein, refers to a non-aromatic saturated monocyclic or multicyclic ring system comprising 3 to about 1 1 ring atoms, wherein from 1 to 4 of the ring atoms are independently O, S, N or Si, and the remainder of the ring atoms are carbon atoms. A heterocycloalkyl group can be joined via a ring carbon, ring silicon atom or ring nitrogen atom. In one embodiment, a heterocycloalkyl group is monocyclic and has from about 3 to about 7 ring atoms. In another embodiment, a heterocycloalkyl group is monocyclic has from about 4 to about 7 ring atoms. In another embodiment, a heterocycloalkyl group is bicyclic and has from about 7 to about 1 1 ring atoms. In still another embodiment, a heterocycloalkyl group is monocyclic and has 5 or 6 ring atoms. In one embodiment, a heterocycloalkyl group is monocyclic. In another embodiment, a heterocycloalkyl group is bicyclic. There are no adjacent oxygen and/or sulfur atoms present in the ring system. Any -NH group in a heterocycloalkyl ring may exist protected such as, for example, as an -N(BOC), -N(Cbz), -N(Tos) group and the like; such protected heterocycloalkyl groups are considered part of this invention. The term "heterocycloalkyl" also encompasses a heterocycloalkyl group, as defined above, which is fused to an aryl (e.g. , benzene) or heteroaryl ring. A heterocycloalkyl group can be optionally substituted by one or more "ring system substituents" which may be the same or different, and are as defined herein below. The nitrogen or sulfur atom of the heterocycloalkyl can be optionally oxidized to the corresponding N-oxide, S-oxide or S,S-dioxide. Non-limiting examples of monocyclic heterocycloalkyl rings include oxetanyl, piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, 1 ,4-dioxanyl, tetrahydro furanyl, tetrahydrothiophenyl, delta-lactam, delta-lactone and the like, and all isomers thereof. A ring carbon atom of a heterocycloalkyl group may be functionalized as a carbonyl group. An illustrative example of such a heterocycloalkyl group is:

In one embodiment, a heterocycloalkyl group is a 5-membered monocyclic heterocycloalkyl. In another embodiment, a heterocycloalkyl group is a 6-membered monocyclic heterocycloalkyl. The term "3 to 6-membered monocyclic heterocycloalkyl" refers to a monocyclic heterocycloalkyl group having from 3 to 6 ring atoms. The term "4 to 7- membered monocyclic heterocycloalkyl" refers to a monocyclic heterocycloalkyl group having from 4 to 7 ring atoms. The term "7 to 11-membered bicyclic heterocycloalkyl" refers to a bicyclic heterocycloalkyl group having from 7 to 11 ring atoms. Unless otherwise indicated, a heterocycloalkyl group is unsubstituted.

Examples of "ring system substituents," include, but are not limited to, alkyl, alkenyl, alkynyl, aryl, heteroaryl, -alkylene-aryl, -arylene-alkyl, -alkylene-heteroaryl, - alkenylene-heteroaryl, -alkynylene-heteroaryl, -OH, hydroxyalkyl, haloalkyl, -O-alkyl, -O- haloalkyl, -alkylene-O-alkyl, -O-aryl, -O-alkylene-aryl, acyl, -C(0)-aryl, halo, -N0 2 , -CN, -SF 5 , -C(0)OH, -C(0)0-alkyl, -C(0)0-aryl, -C(0)0-alkylene-aryl, -S(0)-alkyl, -S(0) 2 -alkyl, -S(O)- aryl, -S(0) 2 -aryl, -S(0)-heteroaryl, -S(0) 2 -heteroaryl, -S-alkyl, -S-aryl, -S-heteroaryl, -S- alkylene-aryl, -S-alkylene-heteroaryl, -S(0) 2 -alkylene-aryl, -S(0) 2 -alkylene-heteroaryl, - Si(alkyl) 2 , -Si(aryl) 2 , -Si(heteroaryl) 2 , -Si(alkyl)(aryl), -Si(alkyl)(cycloalkyl), - Si(alkyl)(heteroaryl), cycloalkyl, heterocycloalkyl, -0-C(0)-alkyl, -0-C(0)-aryl, -O-C(O)- cycloalkyl, -C(=N-CN)-NH 2 , -C(=NH)-NH 2 , -C(=NH)-NH(alkyl), -N(Yi)(Y 2 ), -alkylene- N(Yi)(Y 2 ), -C(0)N(Yi)(Y 2 ) and -S(0) 2 N(Yi)(Y 2 ), wherein Yi and Y 2 can be the same or different and are independently selected from the group consisting of hydrogen, alkyl, aryl, cycloalkyl, and -alkylene-aryl. "Ring system substituent" may also mean a single moiety which simultaneously replaces two available hydrogens on two adjacent carbon atoms (one H on each carbon) on a ring system. Examples of such moiety are methylenedioxy, ethylenedioxy, - C(CH 3 ) 2 - and the like which form moieties such as, for example:

The term "substituted" means that one or more hydrogens on the designated atom is replaced with a selection from the indicated group, provided that the designated atom's normal valency under the existing circumstances is not exceeded, and that the substitution results in a stable compound. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds. By "stable compound' or "stable structure" is meant a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.

The term "in substantially purified form," as used herein, refers to the physical state of a compound after the compound is isolated from a synthetic process (e.g., from a reaction mixture), a natural source, or a combination thereof. The term "in substantially purified form," also refers to the physical state of a compound after the compound is obtained from a purification process or processes described herein or well-known to the skilled artisan (e.g., chromatography, recrystallization and the like), in sufficient purity to be characterizable by standard analytical techniques described herein or well-known to the skilled artisan.

It should also be noted that any carbon as well as heteroatom with unsatisfied valences in the text, schemes, examples and tables herein is assumed to have the sufficient number of hydrogen atom(s) to satisfy the valences.

When a functional group in a compound is termed "protected", this means that the group is in modified form to preclude undesired side reactions at the protected site when the compound is subjected to a reaction. Suitable protecting groups will be recognized by those with ordinary skill in the art as well as by reference to standard textbooks such as, for example, T. W. Greene et al, Protective Groups in Organic Synthesis (1991), Wiley, New York.

When any substituent or variable (e.g., alkyl, R 1 , R 7 , etc.) occurs more than one time in any constituent or in Formula (I), its definition on each occurrence is independent of its definition at every other occurrence, unless otherwise indicated.

As used herein, the term "composition" is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, from combination of the specified ingredients in the specified amounts.

Prodrugs and solvates of the compounds of the invention are also contemplated herein. A discussion of prodrugs is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems (1987) 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, (1987) Edward B. Roche, ed., American Pharmaceutical Association and

Pergamon Press. The term "prodrug" means a compound (e.g., a drug precursor) that is transformed in vivo to provide a Spirocyclic Heterocycle Compound or a pharmaceutically acceptable salt of the compound. The transformation may occur by various mechanisms (e.g., by metabolic or chemical processes), such as, for example, through hydrolysis in blood. For example, if a Spirocyclic Heterocycle Compound or a pharmaceutically acceptable salt, hydrate or solvate of the compound contains a carboxylic acid functional group, a prodrug can comprise an ester formed by the replacement of the hydrogen atom of the acid group with a group such as, for example, (Ci-Cg)alkyl, (C 2 -Ci 2 )alkanoyloxymethyl, l-(alkanoyloxy)ethyl having from 4 to 9 carbon atoms, 1 -methyl- l-(alkanoyloxy)-ethyl having from 5 to 10 carbon atoms,

alkoxycarbonyloxymethyl having from 3 to 6 carbon atoms, l-(alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms, 1 -methyl- l-(alkoxycarbonyloxy)ethyl having from 5 to 8 carbon atoms, N-(alkoxycarbonyl)aminomethyl having from 3 to 9 carbon atoms, 1-(N-

(alkoxycarbonyl)amino)ethyl having from 4 to 10 carbon atoms, 3-phthalidyl, 4-crotonolactonyl, gamma-butyrolacton-4-yl, di-N,N-(Ci-C 2 )alkylamino(C 2 -C 3 )alkyl (such as β- dimethylaminoethyl), carbamoyl-(Ci-C 2 )alkyl, N,N-di (Ci-C 2 )alkylcarbamoyl-(Ci-C 2 )alkyl and piperidino-, pyrrolidino- or morpholino(C 2 -C 3 )alkyl, and the like.

Similarly, if a Spirocyclic Heterocycle Compound contains an alcohol functional group, a prodrug can be formed by the replacement of one or more of the hydrogen atoms of the alcohol groups with a group such as, for example, (Ci-C 6 )alkanoyloxymethyl, l-((Ci- C 6 )alkanoyloxy)ethyl, 1 -methyl- 1 -((C i -C 6 )alkanoyloxy)ethyl, (C i-C 6 )alkoxycarbonyloxymethyl, N-(Ci-C 6 )alkoxycarbonylaminomethyl, succinoyl, (Ci-C 6 )alkanoyl, a-amino(Ci-C 4 )alkyl, a- amino(Ci-C 4 )alkylene-aryl, arylacyl and a-aminoacyl, or α-aminoacyl-a-aminoacyl, where each a-aminoacyl group is independently selected from the naturally occurring L-amino acids, or glycosyl (the radical resulting from the removal of a hydroxyl group of the hemiacetal form of a carbohydrate).

If a Spirocyclic Heterocycle Compound incorporates an amine functional group, a prodrug can be formed by the replacement of a hydrogen atom in the amine group with a group such as, for example, R-carbonyl-, RO-carbonyl-, NRR'-carbonyl- wherein R and R' are each independently (Ci-Cio)alkyl, (C 3 -C7) cycloalkyl, benzyl, a natural α-aminoacyl, - C(OH)C(0)OY 1 wherein Y 1 is H, (Ci-C 6 )alkyl or benzyl, -C(OY 2 )Y 3 wherein Y 2 is (C 1 -C4) alkyl and Y 3 is (Ci-C 6 )alkyl; carboxy (Ci-C 6 )alkyl; amino(Ci-C 4 )alkyl or mono-N- or di-N,N-(C - C 6 )alkylaminoalkyl; -C(Y 4 )Y 5 wherein Y 4 is H or methyl and Y 5 is mono-N- or di-N,N-(Ci- C 6 )alkylamino morpholino; piperidin-l-yl or pyrrolidin-l-yl, and the like.

Pharmaceutically acceptable esters of the present compounds include the following groups: (1) carboxylic acid esters obtained by esterification of the hydroxy group of a hydroxyl compound, in which the non-carbonyl moiety of the carboxylic acid portion of the ester grouping is selected from straight or branched chain alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, t-butyl, sec-butyl or n-butyl), alkoxyalkyl (e.g., methoxymethyl), aralkyl (e.g., benzyl), aryloxyalkyl (for example, phenoxymethyl), aryl (e.g., phenyl optionally substituted withours, for example, halogen, Ci_ 4 alkyl, -0-(Ci_ 4 alkyl) or amino); (2) sulfonate esters, such as alkyl- or aralkylsulfonyl (for example, methanesulfonyl); (3) amino acid esters, including those

corresponding to both natural and non-natural amino acids (e.g., L-valyl or L-isoleucyl); (4) phosphonate esters and (5) mono-, di- or triphosphate esters. The phosphate esters may be further esterified by, for example, a Ci_ 2 o alcohol or reactive derivative thereof, or by a 2,3-di (C 6 _ 2 4)acyl glycerol.

One or more compounds of the invention may exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and it is intended that the invention embrace both solvated and unsolvated forms. "Solvate" means a physical association of a compound of this invention with one or more solvent molecules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain instances the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. "Solvate" encompasses both solution-phase and isolatable solvates. Non- limiting examples of solvates include ethanolates, methanolates, and the like. A "hydrate" is a solvate wherein the solvent molecule is water.

One or more compounds of the invention may optionally be converted to a solvate.

Preparation of solvates is generally known. Thus, for example, M. Caira et al, J. Pharmaceutical Sci., 93(3), 601-611 (2004) describe the preparation of the solvates of the antifungal fluconazole in ethyl acetate as well as from water. Similar preparations of solvates, hemisolvate, hydrates and the like are described by E. C. van Tonder et al, AAPS PharmSciTechours. , 5(1), article 12 (2004); and A. L. Bingham et al, Chem. Commun., 603-604 (2001). A typical, non-limiting, process involves dissolving the inventive compound in desired amounts of the desired solvent (organic or water or mixtures thereof) at a higher than room temperature, and cooling the solution at a rate sufficient to form crystals which are then isolated by standard methods.

Analytical techniques such as, for example IR spectroscopy, show the presence of the solvent (or water) in the crystals as a solvate (or hydrate).

The Spirocyclic Heterocycle Compounds can form salts which are also within the scope of this invention. Reference to a Spirocyclic Heterocycle Compound herein is understood to include reference to salts thereof, unless otherwise indicated. The term "salt(s)", as employed herein, denotes acidic salts formed with inorganic and/or organic acids, as well as basic salts formed with inorganic and/or organic bases. In addition, when a Spirocyclic Heterocycle

Compound contains both a basic moiety, such as, but not limited to a pyridine or imidazole, and an acidic moiety, such as, but not limited to a carboxylic acid, zwitterions ("inner salts") may be formed and are included within the term "salt(s)" as used herein. In one embodiment, the salt is a pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salt. In another embodiment, the salt is other than a pharmaceutically acceptable salt. Salts of the Compounds of Formula (I) may be formed, for example, by reacting a Spirocyclic Heterocycle Compound with an amount of acid or base, such as an equivalent amount, in a medium such as one in which the salt precipitates or in an aqueous medium followed by lyophilization.

Exemplary acid addition salts include acetates, ascorbates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, fumarates, hydrochlorides, hydrobromides, hydroiodides, lactates, maleates, methanesulfonates, naphthalenesulfonates, nitrates, oxalates, phosphates, propionates, salicylates, succinates, sulfates, tartarates, thiocyanates, toluenesulfonates (also known as tosylates) and the like.

Additionally, acids which are generally considered suitable for the formation of pharmaceutically useful salts from basic pharmaceutical compounds are discussed, for example, by P. Stahl et al, Camille G. (eds.) Handbook of Pharmaceutical Salts. Properties, Selection and Use. (2002) Zurich: Wiley- VCH; S. Berge et al, Journal of Pharmaceutical Sciences (1977) 66(1) 1-19; P. Gould, International J. of Pharmaceutics (1986) 33 201-217; Anderson et al, The Practice of Medicinal Chemistry (1996), Academic Press, New York; and in The Orange Book (Food & Drug Administration, Washington, D.C. on their website). These disclosures are incorporated herein by reference thereto.

Exemplary basic salts include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases (for example, organic amines) such as dicyclohexylamine, t-butyl amine, choline, and salts with amino acids such as arginine, lysine and the like. Basic nitrogen- containing groups may be quarternized with agents such as lower alkyl halides (e.g., methyl, ethyl, and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, and dibutyl sulfates), long chain halides (e.g., decyl, lauryl, and stearyl chlorides, bromides and iodides), aralkyl halides (e.g., benzyl and phenethyl bromides), and others.

All such acid salts and base salts are intended to be pharmaceutically acceptable salts within the scope of the invention and all acid and base salts are considered equivalent to the free forms of the corresponding compounds for purposes of the invention. Diastereomeric mixtures can be separated into their individual diastereomers on the basis of their physical chemical differences by methods well-known to those skilled in the art, such as, for example, by chromatography and/or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereomers to the corresponding pure enantiomers. Sterochemically pure compounds may also be prepared by using chiral starting materials or by employing salt resolution techniques. Also, some of the Spirocyclic Heterocycle Compounds may be

atropisomers (e.g., substituted biaryls) and are considered as part of this invention. Enantiomers can also be directly separated using chiral chromatographic techniques.

It is also possible that the Spirocyclic Heterocycle Compounds may exist in different tautomeric forms, and all such forms are embraced within the scope of the invention. For example, all keto-enol and imine-enamine forms of the compounds are included in the invention.

All stereoisomers (for example, geometric isomers, optical isomers and the like) of the present compounds (including those of the salts, solvates, hydrates, esters and prodrugs of the compounds as well as the salts, solvates and esters of the prodrugs), such as those which may exist due to asymmetric carbons on various substituents, including enantiomeric forms (which may exist even in the absence of asymmetric carbons), rotameric forms, atropisomers, and diastereomeric forms, are contemplated within the scope of this invention. If a Spirocyclic Heterocycle Compound incorporates a double bond or a fused ring, both the cis- and trans-forms, as well as mixtures, are embraced within the scope of the invention.

Individual stereoisomers of the compounds of the invention may, for example, be substantially free of other isomers, or may be admixed, for example, as racemates or with all other, or other selected, stereoisomers. The chiral centers of the present invention can have the S or R configuration as defined by the IUPAC 1974 Recommendations. The use of the terms "salt", "solvate", "ester", "prodrug" and the like, is intended to apply equally to the salt, solvate, ester and prodrug of enantiomers, stereoisomers, rotamers, tautomers, racemates or prodrugs of the inventive compounds.

In the Compounds of Formula (I), the atoms may exhibit their natural isotopic abundances, or one or more of the atoms may be artificially enriched in a particular isotope having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number predominantly found in nature. The present invention is meant to include all suitable isotopic variations of the compounds of generic Formula I. For example, different isotopic forms of hydrogen (H) include protium (1H) and deuterium ( 2 H). Protium is the predominant hydrogen isotope found in nature. Enriching for deuterium may provide certain therapeutic advantages, such as increasing in vivo half-life or reducing dosage requirements, or may provide a compound useful as a standard for characterization of biological samples.

Isotopically-enriched Compounds of Formula (I) can be prepared without undue experimentation by conventional techniques well known to those skilled in the art or by processes analogous to those described in the Schemes and Examples herein using appropriate isotopically-enriched reagents and/or intermediates. In one embodiment, a Compound of Formula (I) has one or more of its hydrogen atoms replaced with deuterium.

The Spirocyclic Heterocycle Compounds are useful in human and veterinary medicine for treating or preventing HIV infection in a subject. In one embodiment, the

Spirocyclic Heterocycle Compounds can be inhibitors of HIV viral replication. In a specific embodiment, the Spirocyclic Heterocycle Compounds are inhibitors of HIV- 1. Accordingly, the Spirocyclic Heterocycle Compounds are useful for treating HIV infections and AIDS. In accordance with the invention, the Spirocyclic Heterocycle Compounds can be administered to a subject in need of treatment or prevention of HIV infection.

Accordingly, in one embodiment, the invention provides methods for treating HIV infection in a subject comprising administering to the subject an effective amount of at least one Spirocyclic Heterocycle Compound or a pharmaceutically acceptable salt thereof. In a specific embodiment, the present invention provides methods for treating AIDS in a subject comprising administering to the subject an effective amount of at least one Spirocyclic

Heterocycle Compound or a pharmaceutically acceptable salt thereof.

List of Abbreviations

Anal. = analytical

w-BuLi = w-butyl lithium

m-CPBA = 3-chloroperoxybenzoic acid

DCM = dichloromethane

DEA = diethylamine

DIPEA = N,N-diisopropylethylamine

DMF = dimethylformamide

ESI = electrospray ionization EtOAc = ethyl acetate

EtOH = ethanol

HPLC = high-pressure liquid chromatography

IPA = zsopropyl alcohol

IPAc = zsopropyl acetate

KF = Karl-Fischer titration (to determine water content)

KOi-Bu = potassium tert-butoxide

LCMS = liquid chromatography-mass spectrometry

LiHMDS = lithum hexamethyl silazane

MeCN = acetonitrile

MeOH" = methanol

MPa = milipascal

MS = mass spectroscopy

MTBE = methyl tert-butyl ether

NHS = normal human serum

NMR = nuclear magnetic resonance spectroscopy

Piv = pivalate, 2,2-dimethylpropanoyl

Pd/C = palladium on carbon

rt = room temperature

SFC = supercritical fluid chromatography

TFA = trifluoroacetic acid

TLC = thin-layer chromatography

p-TsOH = para-toluene sulfonic acid

THF = tetrahydrofuran

The Compounds of Formula (I)

The present invention provides Spirocyclic Heterocycle Compounds of Formula

(I) and pharmaceutically acceptable salts thereof, wherein A, B, X, Y, R 1 , R 2 and R 11 are defined above for the Compounds of Formula (I).

In one embodiment, A is 5 or 6-membered monocyclic heterocycle.

In another embodiment, A is 5-membered monocyclic heterocycle.

In one embodiment, X is -CH 2 -.

In another embodiment, X is -CH(CH 3 )-.

In one embodiment, R 11 is H.

In another embodiment, R 11 is C 3 -C 7 cycloalkyl.

In another embodiment, R 11 is -(C 1 -C4 alkylene)-0-(Ci-C6 alkyl).

In another embodiment, R 11 is -CH 2 OCH 3 .

In one embodiment, the compounds of formula (I) have the formula (la):

(la)

or a pharmaceutically acceptable salt or prodrug thereof,

wherein:

A is: -NHC(O)- or:

B is a 5 or 6-membered heterocycloalkyl;

Y is -CH 2 - or -N(CH 3 )-;

R 1 is selected from Ci-C 6 alkyl and -(C 2 -C 4 alkylene)-0-(Ci-C 6 alkyl);

R 2 represents up to 2 optional substituents, each independently selected from halo.

In one embodiment, for the compounds of formula (I) or (la), A is -NHC(O)-. In another embodiment, embodiment, for the compounds of formula (I) or (la), A is: In one embodiment, for the compounds of formula (I) or (la), B is 5-membered heterocycloalkyl.

In another embodiment, for the compounds of formula (I) or (la), B is 6- membered heterocycloalkyl.

In another embodiment, for the compounds of formula (I) or (la), B is tetrahydrofuranyl.

In still another embodiment, for the compounds of formula (I) or (la), B is tetr ahy dropyrany 1.

In another embodiment, for the compounds of formula (I) or (la), B is piperidinyl, optionally substituted on the ring nitrogen atom with with -C(0)OR 5 , -C(0)R 5 , -S(0) 2 -(Ci-C 6 alkyl) or -S(0) 2 NH-(Ci-C 6 alkyl).

In one embodiment, for the compounds of formula (I) or (la), Y is -CH 2 -.

In another embodiment, for the compounds of formula (I) or (la), Y is -N(CH 3 )-. In one embodiment, for the compounds of formula (I) or (la), R 1 is Ci-C 6 alkyl. In another embodiment, for the compounds of formula (I) or (la), R 1 is -(C 2 -C 4 alkylene)-0-(Ci-C 6 alkyl).

In another embodiment, for the compounds of formula (I) or (la), R 1 is selected from methyl, ethyl and n-propyl.

In still another embodiment, for the compounds of formula (I) or (la), R 1 is -CH 2 CH 2 OCH 3 .

In one embodiment, for the compounds of formula (I) or (la), each occurrence of

R 2 is halo.

In another embodiment, for the compounds of formula (I) or (la), R 2 represents 2 fluoro groups.

In another embodiment, for the compounds of formula (I) or (la), R 2 represents 2 fluoro groups in the ortho and para positions.

In one embodiment, variables A, B, X, Y, R 1 , R 2 and R 11 for the Compounds of Formula (I) are selected independently of each other.

In another embodiment, the Compounds of Formula (I) are in substantially purified form.

Other embodiments of the present invention include the following: (a) A pharmaceutical composition comprising an effective amount of a Compound of Formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

(b) The pharmaceutical composition of (a), further comprising a second therapeutic agent selected from the group consisting of HIV antiviral agents, immunomodulators, and anti-infective agents.

(c) The pharmaceutical composition of (b), wherein the HIV antiviral agent is an antiviral selected from the group consisting of HIV protease inhibitors, HIV integrase inhibitors, nucleoside reverse transcriptase inhibitors, CCR5 co-receptor antagonists and non- nucleoside reverse-transcriptase inhibitors.

(d) A pharmaceutical combination that is (i) a Compound of Formula (I) and (ii) a second therapeutic agent selected from the group consisting of HIV antiviral agents, immunomodulators, and anti-infective agents; wherein the Compound of Formula (I) and the second therapeutic agent are each employed in an amount that renders the combination effective for inhibiting HIV replication, or for treating HIV infection and/or reducing the likelihood or severity of symptoms of HIV infection.

(e) The combination of (d), wherein the HIV antiviral agent is an antiviral selected from the group consisting of HIV protease inhibitors, HIV integrase inhibitors, nucleoside reverse transcriptase inhibitors, CCR5 co-receptor antagonists and non-nucleoside reverse-transcriptase inhibitors.

(f) A method of inhibiting HIV replication in a subject in need thereof which comprises administering to the subject an effective amount of a Compound of Formula (I).

(g) A method of treating HIV infection and/or reducing the likelihood or severity of symptoms of HIV infection in a subject in need thereof which comprises

administering to the subject an effective amount of a Compound of Formula (I).

(h) The method of (g), wherein the Compound of Formula (I) is administered in combination with an effective amount of at least one second therapeutic agent selected from the group consisting of HIV antiviral agents, immunomodulators, and anti-infective agents.

(i) The method of (h), wherein the HIV antiviral agent is an antiviral selected from the group consisting of HIV protease inhibitors, HIV integrase inhibitors, nucleoside reverse transcriptase inhibitors, CCR5 co-receptor antagonists and non-nucleoside reverse- transcriptase inhibitors. (j) A method of inhibiting HIV replication in a subject in need thereof which comprises administering to the subject the pharmaceutical composition of (a), (b) or (c) or the combination of (d) or (e).

(k) A method of treating HIV infection and/or reducing the likelihood or severity of symptoms of HIV infection in a subject in need thereof which comprises

administering to the subject the pharmaceutical composition of (a), (b) or (c) or the combination of (d) or (e).

The present invention also includes a compound of the present invention for use (i) in, (ii) as a medicament for, or (iii) in the preparation of a medicament for: (a) medicine, (b) inhibiting HIV replication or (c) treating HIV infection and/or reducing the likelihood or severity of symptoms of HIV infection. In these uses, the compounds of the present invention can optionally be employed in combination with one or more second therapeutic agents selected from HIV antiviral agents, anti-infective agents, and immunomodulators.

Additional embodiments of the invention include the pharmaceutical compositions, combinations and methods set forth in (a)-(k) above and the uses set forth in the preceding paragraphours, wherein the compound of the present invention employed therein is a compound of one of the embodiments, aspects, classes, sub-classes, or features of the

compounds described above. In all of these embodiments, the compound may optionally be used in the form of a pharmaceutically acceptable salt or hydrate as appropriate. It is understood that references to compounds would include the compound in its present form as well as in different forms, such as polymorphs, solvates and hydrates, as applicable.

It is further to be understood that the embodiments of compositions and methods provided as (a) through (k) above are understood to include all embodiments of the compounds, including such embodiments as result from combinations of embodiments.

The Compounds of Formula (I) may be referred to herein by chemical structure and/or by chemical name. In the instance that both the structure and the name of a Compound of Formula (I) are provided and a discrepancy is found to exist between the chemical structure and the corresponding chemical name, it is understood that the chemical structure will predominate.

Non- limiting examples of the Compounds of Formula (I) include compounds 5- 61 as set forth below, and pharmaceutically acceptable salts thereof.

Treatment or Prevention of HIV Infection

The Spirocyclic Heterocycle Compounds are useful in the inhibition of HIV, the inhibition of HIV integrase, the treatment of HIV infection and/or reduction of the likelihood or severity of symptoms of HIV infection and the inhibition of HIV viral replication and/or HIV viral production in a cell-based system. For example, the Spirocyclic Heterocycle Compounds are useful in treating infection by HIV after suspected past exposure to HIV by such means as blood transfusion, exchange of body fluids, bites, accidental needle stick, or exposure to subject blood during surgery or other medical procedures.

Accordingly, in one embodiment, the invention provides methods for treating HIV infection in a subject, the methods comprising administering to the subject an effective amount of at least one Spirocyclic Heterocycle Compound or a pharmaceutically acceptable salt thereof. In a specific embodiment, the amount administered is effective to treat or prevent infection by HIV in the subject. In another specific embodiment, the amount administered is effective to inhibit HIV viral replication and/or viral production in the subject. In one embodiment, the HIV infection has progressed to AIDS.

The Spirocyclic Heterocycle Compounds are also useful in the preparation and execution of screening assays for antiviral compounds. For example the Spirocyclic Heterocycle Compounds are useful for identifying resistant HIV cell lines harboring mutations, which are excellent screening tools for more powerful antiviral compounds. Furthermore, the Spirocyclic Heterocycle Compounds are useful in establishing or determining the binding site of other antivirals to the HIV Integrase.

The compositions and combinations of the present invention can be useful for treating a subject suffering from infection related to any HIV genotype.

Combination Therapy

In another embodiment, the present methods for treating or preventing HIV infection can further comprise the administration of one or more additional therapeutic agents which are not Spirocyclic Heterocycle Compounds.

In one embodiment, the additional therapeutic agent is an antiviral agent.

In another embodiment, the additional therapeutic agent is an immunomodulatory agent, such as an immunosuppressive agent.

Accordingly, in one embodiment, the present invention provides methods for treating a viral infection in a subject, the method comprising administering to the subject: (i) at least one Spirocyclic Heterocycle Compound (which may include two or more different Spirocyclic Heterocycle Compounds), or a pharmaceutically acceptable salt thereof, and (ii) at least one additional therapeutic agent that is other than a Spirocyclic Heterocycle Compound, wherein the amounts administered are together effective to treat or prevent a viral infection. When administering a combination therapy of the invention to a subject, therapeutic agents in the combination, or a pharmaceutical composition or compositions comprising therapeutic agents, may be administered in any order such as, for example, sequentially, concurrently, together, simultaneously and the like. The amounts of the various actives in such combination therapy may be different amounts (different dosage amounts) or same amounts (same dosage amounts). Thus, for non-limiting illustration purposes, a

Spirocyclic Heterocycle Compound and an additional therapeutic agent may be present in fixed amounts (dosage amounts) in a single dosage unit (e.g., a capsule, a tablet and the like).

In one embodiment, the at least one Spirocyclic Heterocycle Compound is administered during a time when the additional therapeutic agent(s) exert their prophylactic or therapeutic effect, or vice versa.

In another embodiment, the at least one Spirocyclic Heterocycle Compound and the additional therapeutic agent(s) are administered in doses commonly employed when such agents are used as monotherapy for treating a viral infection.

In another embodiment, the at least one Spirocyclic Heterocycle Compound and the additional therapeutic agent(s) are administered in doses lower than the doses commonly employed when such agents are used as monotherapy for treating a viral infection.

In still another embodiment, the at least one Spirocyclic Heterocycle Compound and the additional therapeutic agent(s) act synergistically and are administered in doses lower than the doses commonly employed when such agents are used as monotherapy for treating a viral infection.

In one embodiment, the at least one Spirocyclic Heterocycle Compound and the additional therapeutic agent(s) are present in the same composition. In one embodiment, this composition is suitable for oral administration. In another embodiment, this composition is suitable for intravenous administration. In another embodiment, this composition is suitable for subcutaneous administration. In still another embodiment, this composition is suitable for parenteral administration.

Viral infections and virus-related disorders that can be treated or prevented using the combination therapy methods of the present invention include, but are not limited to, those listed above.

In one embodiment, the viral infection is HIV infection.

In another embodiment, the viral infection is AIDS.

The at least one Spirocyclic Heterocycle Compound and the additional therapeutic agent(s) can act additively or synergistically. A synergistic combination may allow the use of lower dosages of one or more agents and/or less frequent administration of one or more agents of a combination therapy. A lower dosage or less frequent administration of one or more agents may lower toxicity of therapy without reducing the efficacy of therapy.

In one embodiment, the administration of at least one Spirocyclic Heterocycle Compound and the additional therapeutic agent(s) may inhibit the resistance of a viral infection to these agents.

As noted above, the present invention is also directed to use of a compound of Formula I with one or more anti-HIV agents. An "anti-HIV agent" is any agent which is directly or indirectly effective in the inhibition of HIV reverse transcriptase or another enzyme required for HIV replication or infection, the treatment or prophylaxis of HIV infection, and/or the treatment, prophylaxis or delay in the onset or progression of AIDS. It is understood that an anti-HIV agent is effective in treating, preventing, or delaying the onset or progression of HIV infection or AIDS and/or diseases or conditions arising therefrom or associated therewith. For example, the compounds of this invention may be effectively administered, whether at periods of pre-exposure and/or post-exposure, in combination with effective amounts of one or more anti- HIV agents selected from HIV antiviral agents, imunomodulators, antiinfectives, or vaccines useful for treating HIV infection or AIDS. Suitable HIV antivirals for use in combination with the compounds of the present invention include, for example, those listed in Table A as follows:

Table A

Name Type

abacavir, ABC, Ziagen® nRTI abacavir +lamivudine, Epzicom® nRTI abacavir + lamivudine + zidovudine, Trizivir® nRTI amprenavir, Agenerase® PI atazanavir, Reyataz® PI

AZT, zidovudine, azidothymidine, Retrovir® nRTI darunavir, Prezista® PI ddC, zalcitabine, dideoxycytidine, Hivid® nRTI ddl, didanosine, dideoxyinosine, Videx® nRTI ddl (enteric coated), Videx EC® nRTI delavirdine, DLV, Rescriptor® nnRTI

Dolutegravir Inl efavirenz, EFV, Sustiva®, Stocrin® nnRTI efavirenz + emtricitabine + tenofovir DF, Atripla® nnRTI + nRTI

Elvitegravir Inl emtricitabine, FTC, Emtriva® nRTI emtricitabine + tenofovir DF, Truvada® nRTI emvirine, Coactinon® nnRTI enfuvirtide, Fuzeon® FI enteric coated didanosine, Videx EC® nRTI etravirine, TMC-125 nnRTI fosamprenavir calcium, Lexiva® PI indinavir, Crixivan® PI lamivudine, 3TC, Epivir® nRTI lamivudine + zidovudine, Combivir® nRTI lopinavir PI lopinavir + ritonavir, Kaletra® PI maraviroc, Selzentry® EI nelfmavir, Viracept® PI nevirapine, NVP, Viramune® nnRTI raltegravir, MK-0518, Isentress® Inl rilpivirine, TMC-278 nnRTI ritonavir, Norvir® PI saquinavir, Invirase®, Fortovase® PI stavudine, d4T,didehydrodeoxythymidine, Zerit® nRTI tenofovir DF (DF = disoproxil fumarate), TDF, nRTI Viread®

tipranavir, Aptivus® PI

EI = entry inhibitor; FI = fusion inhibitor; Inl = integrase inhibitor; PI = protease inhibitor; nRTI = nucleoside reverse transcriptase inhibitor; nnRTI = non-nucleoside reverse transcriptase inhibitor. Some of the drugs listed in the table are used in a salt form; e.g., abacavir sulfate, indinavir sulfate, atazanavir sulfate, nelfmavir mesylate. In one embodiment, the one or more anti-HIV drugs are selected from raltegravir, lamivudine, abacavir, ritonavir, dolutegravir, darunavir, atazanavir, emtricitabine, tenofovir, elvitegravir, rilpivirine and lopinavir.

In another embodiment, the compound of formula (I) is used in combination with a single anti-HIV drug which is raltegravir.

In another embodiment, the compound of formula (I) is used in combination with a single anti-HIV drug which is lamivudine.

In still another embodiment, the compound of formula (I) is used in combination with a single anti-HIV drug which is atazanavir.

In another embodiment, the compound of formula (I) is used in combination with a single anti-HIV drug which is darunavir.

In another embodiment, the compound of formula (I) is used in combination with a single anti-HIV drug which is rilpivirine.

In yet another embodiment, the compound of formula (I) is used in combination with a single anti-HIV drug which is dolutegravir.

In another embodiment, the compound of formula (I) is used in combination with a single anti-HIV drug which is elvitegravir.

In one embodiment, the compound of formula (I) is used in combination with two anti-HIV drugs which are lamivudine and abacavir.

In another embodiment, the compound of formula (I) is used in combination with two anti-HIV drugs which are darunavir and raltegravir.

In another embodiment, the compound of formula (I) is used in combination with two anti-HIV drugs which are emtricitabine and tenofovir.

In still another embodiment, the compound of formula (I) is used in combination with two anti-HIV drugs which are atazanavir and raltegravir.

In another embodiment, the compound of formula (I) is used in combination with two anti-HIV drugs which are ritonavir and lopinavir.

In another embodiment, the compound of formula (I) is used in combination with two anti-HIV drugs which are lamivudine and raltegravir.

In one embodiment, the compound of formula (I) is used in combination with three anti-HIV drug which are abacavir, lamivudine and raltegravir.

In another embodiment, the compound of formula (I) is used in combination with three anti-HIV drug which are lopinavir, ritonavir and raltegravir. In one embodiment, the present invention provides pharmaceutical compositions comprising (i) a compound of formula (I) or a pharmaceutically acceptable salt thereof; (ii) a pharmaceutically acceptable carrier; and (iii) one or more additional anti-HIV agents selected from lamivudine, abacavir, ritonavir and lopinavir, or a pharmaceutically acceptable salt thereof, wherein the amounts present of components (i) and (iii) are together effective for the treatment or prophylaxis of infection by HIV or for the treatment, prophylaxis, or delay in the onset or progression of AIDS in the subject in need thereof.

In another embodiment, the present invention provides a method for the treatment or prophylaxis of infection by HIV or for the treatment, prophylaxis, or delay in the onset or progression of AIDS in a subject in need thereof, which comprises administering to the subject (i) a compound of formula (I) or a pharmaceutically acceptable salt thereof and (ii) one or more additional anti-HIV agents selected from lamivudine, abacavir, ritonavir and lopinavir, or a pharmaceutically acceptable salt thereof, wherein the amounts administered of components (i) and (ii) are together effective for the treatment or prophylaxis of infection by HIV or for the treatment, prophylaxis, or delay in the onset or progression of AIDS in the subject in need thereof.

It is understood that the scope of combinations of the compounds of this invention with anti-HIV agents is not limited to the HIV antivirals listed in Table A, but includes in principle any combination with any pharmaceutical composition useful for the treatment or prophylaxis of AIDS. The HIV antiviral agents and other agents will typically be employed in these combinations in their conventional dosage ranges and regimens as reported in the art, including, for example, the dosages described in the Physicians' Desk Reference, Thomson PDR, Thomson PDR, 57 th edition (2003), the 58 th edition (2004), the 59 th edition (2005), and the like. The dosage ranges for a compound of the invention in these combinations are the same as those set forth above.

The doses and dosage regimen of the other agents used in the combination therapies of the present invention for the treatment or prevention of HIV infection can be determined by the attending clinician, taking into consideration the approved doses and dosage regimen in the package insert; the age, sex and general health of the subject; and the type and severity of the viral infection or related disease or disorder. When administered in combination, the Spirocyclic Heterocycle Compound(s) and the other agent(s) can be administered

simultaneously (i.e. , in the same composition or in separate compositions one right after the other) or sequentially. This particularly useful when the components of the combination are given on different dosing schedules, e.g., one component is administered once daily and another component is administered every six hours, or when the preferred pharmaceutical compositions are different, e.g., one is a tablet and one is a capsule. A kit comprising the separate dosage forms is therefore advantageous. Compositions and Administration

When administered to a subject, the Spirocyclic Heterocycle Compounds can be administered as a component of a composition that comprises a pharmaceutically acceptable carrier or vehicle. The present invention provides pharmaceutical compositions comprising an effective amount of at least one Spirocyclic Heterocycle Compound and a pharmaceutically acceptable carrier. In the pharmaceutical compositions and methods of the present invention, the active ingredients will typically be administered in admixture with suitable carrier materials suitably selected with respect to the intended form of administration, i.e., oral tablets, capsules (either solid-filled, semi-solid filled or liquid filled), powders for constitution, oral gels, elixirs, dispersible granules, syrups, suspensions, and the like, and consistent with conventional pharmaceutical practices. For example, for oral administration in the form of tablets or capsules, the active drug component may be combined with any oral non-toxic pharmaceutically acceptable inert carrier, such as lactose, starchours, sucrose, cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, talc, mannitol, ethyl alcohol (liquid forms) and the like. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets and suppositories. Powders and tablets may be comprised of from about 0.5 to about 95 percent inventive composition. Tablets, powders, cachets and capsules can be used as solid dosage forms suitable for oral administration.

Moreover, when desired or needed, suitable binders, lubricants, disintegrating agents and coloring agents may also be incorporated in the mixture. Suitable binders include starchours, gelatin, natural sugars, corn sweeteners, natural and synthetic gums such as acacia, sodium alginate, carboxymethylcellulose, polyethylene glycol and waxes. Among the lubricants there may be mentioned for use in these dosage forms, boric acid, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include starchours, methylcellulose, guar gum, and the like. Sweetening and flavoring agents and preservatives may also be included where appropriate.

Liquid form preparations include solutions, suspensions and emulsions and may include water or water-propylene glycol solutions for parenteral injection.

Liquid form preparations may also include solutions for intranasal administration. Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for either oral or parenteral administration. Such liquid forms include solutions, suspensions and emulsions.

For preparing suppositories, a low melting wax such as a mixture of fatty acid glycerides or cocoa butter is first melted, and the active ingredient is dispersed homogeneously therein as by stirring. The molten homogeneous mixture is then poured into convenient sized molds, allowed to cool and thereby solidify.

Additionally, the compositions of the present invention may be formulated in sustained release form to provide the rate controlled release of any one or more of the

components or active ingredients to optimize therapeutic effects, i.e., antiviral activity and the like. Suitable dosage forms for sustained release include layered tablets containing layers of varying disintegration rates or controlled release polymeric matrices impregnated with the active components and shaped in tablet form or capsules containing such impregnated or encapsulated porous polymeric matrices.

In one embodiment, the one or more Spirocyclic Heterocycle Compounds are administered orally.

In another embodiment, the one or more Spirocyclic Heterocycle Compounds are administered intravenously.

In one embodiment, a pharmaceutical preparation comprising at least one

Spirocyclic Heterocycle Compound is in unit dosage form. In such form, the preparation is subdivided into unit doses containing effective amounts of the active components.

Compositions can be prepared according to conventional mixing, granulating or coating methods, respectively, and the present compositions can contain, in one embodiment, from about 0.1% to about 99% of the Spirocyclic Heterocycle Compound(s) by weight or volume. In various embodiments, the present compositions can contain, in one embodiment, from about 1% to about 70%> or from about 5% to about 60%> of the Spirocyclic Heterocycle Compound(s) by weight or volume.

The compounds of Formula I can be administered orally in a dosage range of 0.001 to 1000 mg/kg of mammal (e.g., human) body weight per day in a single dose or in divided doses. One preferred dosage range is 0.01 to 500 mg/kg body weight per day orally in a single dose or in divided doses. Another preferred dosage range is 0.1 to 100 mg/kg body weight per day orally in single or divided doses. For oral administration, the compositions can be provided in the form of tablets or capsules containing 1.0 to 500 milligrams of the active ingredient, particularly 1, 5, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, and 500 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the subject to be treated. The specific dose level and frequency of dosage for any particular subject may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general healthours, sex, diet, mode and time of administration, rate of excretion, drug

combination, the severity of the particular condition, and the host undergoing therapy.

For convenience, the total daily dosage may be divided and administered in portions during the day if desired. In one embodiment, the daily dosage is administered in one portion. In another embodiment, the total daily dosage is administered in two divided doses over a 24 hour period. In another embodiment, the total daily dosage is administered in three divided doses over a 24 hour period. In still another embodiment, the total daily dosage is administered in four divided doses over a 24 hour period.

The amount and frequency of administration of the Spirocyclic Heterocycle Compounds will be regulated according to the judgment of the attending clinician considering such factors as age, condition and size of the subject as well as severity of the symptoms being treated. The compositions of the invention can further comprise one or more additional therapeutic agents, selected from those listed above herein.

Kits

In one aspect, the present invention provides a kit comprising a therapeutically effective amount of at least one Spirocyclic Heterocycle Compound, or a pharmaceutically acceptable salt or prodrug of said compound and a pharmaceutically acceptable carrier, vehicle or diluent.

In another aspect the present invention provides a kit comprising an amount of at least one Spirocyclic Heterocycle Compound, or a pharmaceutically acceptable salt or prodrug of said compound and an amount of at least one additional therapeutic agent listed above, wherein the amounts of the two or more active ingredients result in a desired therapeutic effect. In one embodiment, the one or more Spirocyclic Heterocycle Compounds and the one or more additional therapeutic agents are provided in the same container. In one embodiment, the one or more Spirocyclic Heterocycle Compounds and the one or more additional therapeutic agents are provided in separate containers.

Methods For Making the Compounds of Formula (I) The Compounds of Formula (I) may be prepared from known or readily prepared starting materials, following methods known to one skilled in the art of organic synthesis.

Methods useful for making the Compounds of Formula (I) are set forth in the Examples below and generalized in Scheme 1 below. Alternative synthetic pathways and analogous structures will be apparent to those skilled in the art of organic synthesis.

The following schemes describe general methods for preparing the compounds of formula (I), which correspond to the 4-pyridotriazines compounds of Formula (I).

A pyranone compound of formula A is reacted with an amine of formula B to provide dihydropyridine compounds of formula C. Base promoted hydrolysis of the ester moiety of C to provide carboxylic acids of formula D, followed by amide coupling of D with an amine compound of formula E provides cyclization precursors of formula F. Acid catalyzed deprotection of the benzyl protecting group, followed by condensation with an aldehyde or ketone of formula G provides the product H.

Scheme 2

A pyranone compound of formula A is reacted with an amine of formula B under a variety of conditions to provide either a compound of formula C or to directly provide a compound of formula D. Compounds of formula C can also be converted to a compound of formula D.

Scheme 3

I

A pyranone compound of formula A is converted to a pyridine of the formula B. Amide coupling with a suitable amine provides a compound of the formula C. An amine transfer reagent is utilized to provide a compound of formula D. Acid catalysis and a reagent of the formula E provides the trazinone of the formula F. Alkylation with a suitable alkyl halide provide compound of the formula G. Halogen transfer then provide a compound of the formula H. Transition methal catalysis with a suitable coupling partner provides a compound of the formula I. Finally, deprotection provides a comlound of the formula J .

Scheme 4

E F G A pyranone compound of formula A is coupled to a suitably functionalized amine B for provide a compound of the general formula C. Amine deprotection and subsequent cyclization provides a compound of the general structure D. Halogen transfer provides a compound of the general structure E. Transition methal catalysis with a suitable coupling partner provides a compound of the formula F. Finally, deprotection provides a comlound of the formula G.

EXAMPLES

General Methods

The compounds described herein can be prepared according to the procedures of the following schemes and examples, using appropriate materials and are further exemplified by the following specific examples. The compounds illustrated in the examples are not, however, to be construed as forming the only genus that is considered as the invention. The examples further illustrate details for the preparation of the compounds of the present invention. Those skilled in the art will readily understand that known variations of the conditions and processes of the following preparative procedures can be used to prepare these compounds. All temperatures are degrees Celsius unless otherwise noted. Mass spectra (MS) were measured by electrospray ion- mass spectroscopy (ESI). 1H NMR spectra were recorded at 400-500 MHz. Compounds described herein were synthesized as a racemic mixture unless otherwise stated in the experimental procedures.

Example 1

Preparation of Intermediate Compound 1-1

1a 1 b 1c

Step A - Synthesis of Intermediate Compound lb

To a solution of compound la (200g, 1.2 mol) in dry methanol (2 L) was added SOCb (424 g, 3.6 mol) under N 2 at 0 °C, then heated under reflux for 6 hours. The reaction mixture was concentrated in vacuo and the resulting residue was dissolved in Ethyl acetate (3 L). The organic phase was washed with NaHC0 3 (2 L><2), brine, dried over Na 2 S0 4 and

concentrated in vacuo to provide compound lb as an oil. Step B - Synthesis of Intermediate Compound lc

To a solution of compound lb (322 g, 2.78mol) in dry THF (2.8 L) was added LiHMDS (2.78 L, 2.78 mol) at -70 °C under N 2 . After stirred at -70 °C for 1 hours, then compound 2 (250 g, 1.39 mol) was added and the mixture was allowed to stir at -70 °C for 1.5 hours. The reaction was quenched with water (2 L) and extracted with Ethyl acetate (8 L><3). The combined organic phase was washed with brine, dried over Na 2 S0 4 , and concentrated in vacuo to provide compound lc as an oil.

Step C - Synthesis of Intermediate Compound Id

A mixture of compound lc (85 g, 0.32 mol) and DMF-DMA (76 g, 0.64 mol) in DMF (200 mL) was heated to 100 °C for 6 hours. The reaction was concentrated in vacuo and the resulting residue was purified using flash chromatography on silica gel (PE:EA=50: 1 to 1 :1) to provide compound Id as a solid.

Step D - Synthesis of Intermediate Compound le

To a solution of compound Id (50 g, 0.16 mol) in THF (300 mL) was added

LiHMDS (190 mL, 0.19 mmol) at -70°C under N 2 . After stirring at -70 °C for 0.5hours, ethyl 2- chloro-2-oxoacetate (25.8 g, 0.19 mol) was added and the mixture was allowed to stir at -70 °C for 1 hour. TLC(PE:EA=1 : 1) showed the reaction was complete. The reaction was quenched with sat. aq. KHS0 4 (250 mL) and extracted with Ethyl acetate (500 mL><3). The combined organic phase was washed with brine, dried over Na 2 S0 4 and concentrated in vacuo to provide the crude product. The resulting residue was diluted with toluene and concentrated, then toluene (500 mL) and triethylamine (50 mL) were added and the mixture was allowed to stir at room temperature for 1 hour. The mixture was concentrated in vacuo and the crude product was purified using flash chromatography on silica gel (PE:EA= 1 :0 to 40: 1) to provide compound le as a solid.

Step E - Synthesis of Intermediate Compound If

To a stirring solution of the compound le (85 g, 0.23 mol) in Ethyl acetate (100 mL) was added HCl/Ethyl acetate (4 N, 920 mL) at 0 °C and the resulting mixture was allowed to stir at room temperature for 1 hour. TLC (PE: EA= 5: 1) showed the reaction was complete. The reaction was concentrated in vacuo and to the resulting residue was added hexane (1 L). The mixture was allowed to stir for 1 h and filtered to provide If as a solid.

Steps F - Synthesis of Compound I -I

To a stirring solution of the compound If (3.3 g, 10.3 mmol) in toluene (70 mL) was added oxalyl chloride (20.6 mL) and DMF (0.2 mL) at 0 °C under N 2 and the resulting mixture was allowed to stir at room temperature for 2 hours. The mixture was concentrated in vacuo and to the resulting residue was added CHCI3 (100 mL) and compound 2e (2.84 g, 15.45 mmol). The resulting mixture was allowed to stir at room temperature overnight. TLC (CH 2 C1 2 : methanol= 10: 1) showed the starting material was consumed. To the reaction mixture was added HC1 (4M, 10 mL in MTBE) the mixture was allowed to stir at room temperature for 2h. To the mixture was added 5% aq. KHS0 4 (200 mL) and extracted with CHC1 2 twice. The organic layer was washed with brine, dried over Na 2 S0 4 and concentrated in vacuo. The crude product was purified using silica gel column (PE:EA=50: 1 to 1 : 1) to provide 1-1 as a solid. 1H NMR (CDCI3, 400 MHz) 9.07 (s, 1H), 7.45 (m, 2H), 7.35-7.25 (m, 5H), 7.03(m, 2H).5.34 (s, 2H),4.39-4.37 (q, 2H), 1.36- 1.32 (t, 3H) LCMS (M + H) = 467.

The intermediate compound set forth in the table below was made using the method described in the Example above and substituting the appropriate reactants and reagents:

Example 2

Preparation of Intermediate Compound 2

Step A - Synthesis of Intermediate Compound 2b

To a solution of compound 2a (100 g, 0.65 mol) in anhydrous dichloromethane (1 L) was added SOCl 2 (200 mL) dropwise at 0 °C under a drying tube charged with CaCl 2 . After the addition, the mixture was heated to reflux and stirred overnight. The reaction was done in 2 batches, which were combined and concentrated in vacuo to provide crude compound 2b as an oil that was used without further purification.

Step B - Synthesis of Intermediate Compound 2c To a solution of BocNHNH 2 (102.9 g, 0.78 mol) and triethylamine (135.4 mL, 0.97 mol) in anhydrous dichloromethane (800 mL) was added a solution of compound 2b (138 g) in anhydrous dichloromethane (500 mL) at 0 °C under drying tube charging with CaCl 2 . The mixture was warmed up to room temperature and stirred for 2 hours. The mixture was quenched with water (1 L). The reaction was done in two batches which were combined. The two phases were separated and the aqueous layer was extracted with dichloromethane (1L><2). The organic layer was washed with water (1L><4), then brine, dried over Na 2 S0 4 , and concentrated in vacuo to provide compound 2c as a solid. Step C - Synthesis of Intermediate Compound 2d

A mixture of compound 2c (50 g, 186.4 mmol) and Lawesson's reagent (75.4 g, 186.4 mmol) in THF (600 mL) was allowed to stir at 60 °C overnight. The mixture was poured into 10 % Na 2 C03 (1000 mL) and stirred at room temperature for 1 hour. The reaction was done in two batches which were combined, and the mixture was extracted with Ethyl acetate (1L><3). The organic layer was washed with brine, dried over Na 2 S0 4 , and concentrated in vacuo to provide compound 2d as a solid.

Step D - Synthesis of Compound 2

A suspension of compound 2d (240 g, 186.4 mmol, crude) in HCl/methanol (1.2 L, 4N) was allowed to stir at room temperature for 1 hour. The mixture was filtered and the filtrate was concentrated in vacuo to provide the HC1 salt of compound 2, which was dissolved into water. The aqueous layer was basified with 10 % Na 2 C03 until pH = 8 and extracted with ethyl acetate (1L <4). The organic layer was washed with brine, dried over Na 2 S0 4 , concentrated in vacuo to provide compound 2 (35 g) as a white solid. 1H NMR (CDC1 3 , 400 MHz) 8.44 (br, 1H), 7.26-7.21 (m, 2H), 7.07-7.02 (m, 2H), 4.83 (br, 2H), 4.05 (s, 2H) LCMS (M + H) = 185.0

Example 3

Preparation of Intermediate Compound 3

3a 3b 3c

Step A - Synthesis of Intermediate Compound 3b

To the solution of compound 3a (200g, 2.2 mol) in dry methanol (2 L) was added SOCb (778 g, 6.6 mol) under N 2 at 0 °C, then heated to reflux for 6 hours. The reaction mixture was concentrated, the resulting residue was dissolved in EtOAc (3 L) and washed with NaHC0 3 (2 L x 2), the organic phase was washed with brine, dried over anhydrous Na 2 S0 4 and

concentrated in vacuo to provide compound 3b as an oil that was used without further

purification.

Step B - Synthesis of Intermediate Compound 3c

A 10 L three-necked round bottom flask equipped with a mechanic stirrer and thermometer was charged with a solution of iert-butyl acetate (1612, 13.9 mol) in dry THF (14 L) and cooled to -70 °C under N 2 . A solution of LiHMDS (13.9 L, 13.9 mol) in THF was added at - 70 °C and the mixture was allowed to stir at -70 °C for 1 hour. A solution of 3b (723 g, 6.95 mol) in THF (1 L) was added to the reaction and stirred at -70 °C for 0.5 hours. The reaction was quenched by the addition of water (10 L) and the mixture was extracted with EtOAc. The combined the organic phase was washed with brine, dried over Na 2 S0 4 and concentrated in vacuo to provide 3c as oil. 1H NMR (400 MHz, CDC1 3 ) δ 4.09 (s, 2H), 3.43 (s, 2H), 3.42 (s, 2H), 1.47 (s, 9H). Step C - Synthesis of Intermediate Compound 3d

A mixture of 3c (564 g, 3 mol) and DMF-DMA (696 g, 6 mol) in toluene (1380L) was heated to 80 °C for 2 hours. The reaction was concentrated to provide 3d as solid that was used without further purification. 1H NMR (400 MHz, CDC1 3 ) δ 7.65 (s, 1H), 4.35 (s, 2H), 3.40 (s, 2H), 3.36-2.87 (m, 6H), 1.48 (s, 9H).

Step D - Synthesis of Intermediate Compound 3e

A solution of 3d (600 g, 2469 mmol) in dry THF (3600 mL) was cooled to -70 °C under N 2 , and treated dropwise over 0.5 h with a solution of LiHMDS (3 L, 3 mol) in THF. The resulting mixture was then treated at -70°C with ethyl 2-chloro-2-oxoacetate (403 g, 2962 mmol) and mixture was aged at -70°C for 1 hour. The reaction was quenched with saturated aqueous KHSO 4 and extracted with EtOAc. The combined organic phase was washed with brine, dried over anhydrous Na 2 S0 4 and concentrated in vacuo to provide the crude product. The resulting residue was diluted with toluene and concentrated in vacuo. Toluene (6 L) and triethylamine (600 mL) were were added and the mixture was allowed to stir at r.t for 1 h and then

concentrated in vacuo. The crude product was purified using column chromatography on silica gel column (PE:EA= 100:1 to 10: 1) to provide 3e as a solid. 1H NMR (400 MHz, CDC1 3 ) δ 8.41 (s, lH),4.43-4.42 (m, 2H), 4.03 (s, 2H),1.57 (s, 9H), 1.43-1.39 (t, J=7.2, 3H). Step E - Synthesis of Intermediate Compound 3f

To a stirred solution of the 3e (140 g, 469 mmol) in EtOAc (140 mL) at 0 °C was added HC1/ EtOAc (4 N, 1400 mL). The resulting mixture was allowed to stir at r.t for 1 h and then concentrated in vacuo. To the resulting residue was added to hexane (1.4 L). The resulting mixture was allowed to stir at room temperature to provide a precipitate. The mixture was filtered and the solid ws collected and dried in vacuo to provide 3f .

Steps F - Synthesis of Compound 3

To a stirred solution of the 3f (10 g, 41.3 mmol) in toluene (200 mL) was added oxalyl chloride (60 mL) and DMF (0.6 mL) at 0 °C under N 2 . The resulting mixture was allowed to stir at r.t for 2 h and then the mixture was concentrated in vacuo. The resulting residue was dissolved in CHC1 3 (300 mL), treated with 4 (12.5 g, 61.9 mmol) and aged at room temperature for 16 hours. The crude product was recystallization with PE and EA to provide 3 as a solid. 1H NMR (400 MHz, CDC1 3 ) δ 9.08 (s, 1H), 7.33-7.27 (m, 1H), 6.89-6.84 (m, 2H), 4.49-4.44 (m, 4H), 4.08 (s, 3H),1.45-1.42 (t, J=7.2, 3H). Example 4

Preparation of Intermediate Compound 4

4c 4

Step A - Synthesis of Intermediate Compound 4a

To a solution of 2-(2,4-difluorophenyl)acetic acid (100 g, 0.58 mol) in anhydrous dichloromethane (1000 mL) was added thionyl chloride (200 mL) at 0 °C. After addition was complete, the mixture was heated at reflux and stirred overnight. The solution was concentrated in vacuo to provide the crude compound 4b as an oil that was used without further purification.

Step B - Synthesis of Intermediate Compound 4b

To a solution of BocNHNH2 (48 g, 0.64 mol) and triethylamine (110 mL, 0.72 mol) in anhydrous dichloromethane (600 mL) was added a solution of crude compound 4b (100 g, 0.53 mol) in anhydrous dichloromethane (1000 mL) at 0 °C. After addition ws complete, the mixture was warmed up to room temperature and stirred for 2 hrs. The mixture was quenched by the addition of water (500 mL). The two phases were separated and the aqueous layer was extracted with dichloromethane (500 mL x 2). The organic layer was washed with water (500 mL x 4), brine, dried over anhydrous Na 2 S0 4 , and concentrated in vacuo to provide 4c as a solid.

Step C - Synthesis of Intermediate Compound 4d

A mixture of compound 4c (50 g, 0.19 mol) and Lawesson's reagent (105 g, 0.25 mol) in THF (600 mL) was allowed to stir at 50 °C for 2 hours. The mixture was then poured into 10% aqueous Na 2 C0 3 (1 L) and extracted with ethyl acetate (1 L x 3). The combined organic extracts was washed with brine (500 mL), dried over anhydrous Na 2 S0 4 and

concentrated in vacuo to provide compound 4d as a solid. Step D - Synthesis of Compound 4

The suspension of 4d (70 g, 23 mmol) in HCl/ methanol (500 mL, 4N) was allowed to stir at room temperature for 1 hour. The mixture was filtered and the filtrate was concentrated in vacuo to provide 3e as the HCl salt. The solid was dissolved in water and the aqueous layer was adjusted to pH = 10 with 10% of aqueous Na 2 C0 3 . The solution was extracted with EtOAc (200 mL*6). The combined organic phase was washed with brine (500 mL), dried over Na 2 S0 4 , and concentrated in vacuo to provide 4 as a solid. 1H NMR (400 MHz, CDC1 3 ) d 8.51 (br, 1H), 7.35-7.37 (d, 1H), 6.84-6.91 (m, 2H), 4.2 (br, 2H), 4.0 (s, 2H).

Step A - Synthesis of Intermediate Compound 5a

Compound lg (3.00 g, 6.43 mmol) in N-methylimidazole (15.0 ml) was treated with ie/t-butyl 1-methylhydrazinecarboxylate (1.43 ml, 9.65 mmol) at room temperature. The mixture was heated at 70 °C for 6 h and then cooled to room temperature and poured into ethyl acetate (50 mL). The combined organic extracts were washed with aqueous citric acid (1 M, 2 x 25 mL), water and brine. The organic layer was dried (Na 2 S0 4 ), filtered and the filtrate was concentrated in vacuo. Purification using column chromatography on silica gel (0 to 100% Ethyl acetate/ heptane) provided compound 5a. 1H NMR (CDC1 3 , 400 MHz) 8.67 (s, 1H), 7.43- 7.40 (m, 2H), 7.36-7.30 (m, 5H), 7.05-7.00 (m, 2H), 5.42-5.35 (bs, 1H), 5.31-5.26 (bs, 1H), 4.45 (s, 2H), 4.31-4.28 (m, 2H), 3.38 (s, 3H), 1.40 (s, 9H), 1.25 (t, 3H) LCMS (M + H) = 595

Step B - Synthesis of Intermediate Compound 5b Compound 5a (0.10 g, 0.17 mmol) in THF (0.50 ml)/Water (0.17 ml)/methanol (0.17 ml) was added sodium hydroxide (0.21 ml, 0.42 mmol) and stirred for 16 h at 25 °C.

Neutralize with 2N HC1 (0.21 ml) and concentrate. The solid was taken up in Cf^CVmethanol and filtered. The filtrate was concentrated in vacuo and purified using preparative reverse-phase HPLC to provide 5b. 1H NMR (CDC1 3 , 400 MHz) 8.78 (s, IH), 7.43-7.41 (m, 2H), 7.34-7.28 (m, 5H), 7.04-7.00 (m, 2H), 5.45 (s, 2H), 4.45 (s, 2H), 3.39 (s, 3H), 1.41 (s, 9H), LCMS (M + H) = 567

Step C - Synthesis of Intermediate Compound 5c

3 -(benzyloxy)- 1 -((tert-butoxycarbonyl)(methyl)amino)-5 -(5 -(4-fluorobenzyl)- l,3,4-thiadiazol-2-yl)-4-oxo-l,4-dihydropyridine-2-carboxyli c acid 5b (0.16 g, 0.29 mmol) in CH 2 CI 2 (1.4 ml) was added l-(chloror-l-pyrrolidinylmethylene)pyrrolidinium

hexafluorophosphate (0.096 g, 0.29 mmol) followed by ethylamine (0.15 ml, 0.29 mmol) and N,N-diisopropylethylamine (0.20 ml, 1.2 mmol). The reaction stirred for 30 minutes and was added another equivalent of PyClu and ethylamine. After stirring for an additional 1 hour, the solution was extracted with CH 2 CI 2 and washed with saturated NaHC0 3 . The organic layer was dried, concentrated in vacuo, and purified using preparative reverse-phase HPLC to provide 5c. 1H NMR (CDC1 3 , 400 MHz) 8.68 (s, IH), 7.38-7.29 (m, 7H), 7.04-7.00 (m, 2H), 5.85 (bs, IH), 5.57 (d, IH), 5.11 (d, IH), 4.45 (s, 2H), 3.43 (s, 3H), 3.27-3.20 (m, 2H), 1.45 (s, 9H), 1.02 (t, 3H) LCMS (M + H) = 594

Step D - Synthesis of Compound 5

Compound 5c (101 mg, 0.170 mmol) and N-carbethoxy-4-piperidone (146 mg,

0.851 mmol) were dissolved in dichloroacetic acid (0.5 ml). The mixture was heated at 95 °C for 8 hours. The mixture was cooled to room temperature, diluted with DMSO (3 mL) and directly purified using preparative reverse-phase HPLC to provide compound 5. LCMS (M + H)

= 557; 1H NMR (500 MHz, acetone): δ 8.82 (s; 1 H); 7.44 (dd; J = 8.35; 5.43 Hz; 2 H); 7.11 (t;

J = 8.73 Hz; 2 H); 4.47 (s; 2 H); 4.21 (br s; 2 H); 4.08 (d; J = 8.13 Hz; 2 H); 3.97 (br s; 2 H);

3.83 (dq; J = 14.5; 7.1 Hz; 1 H); 3.65 (dq; J = 14.5; 7.1 Hz; 1 H); 3.29 (br s; 3 H); 2.98 (s; 3 H); 1.88-1.94 (m; 1 H); 1.29 (t; J = 7.12 Hz; 3 H); 1.21 (s; 3 H).

The following compounds of the present invention were made using the method described in the Example above and substituting the appropriate reactants and reagents.

Example 6

Preparation of Compound 32

Step A- Synthesis of Intermediate Compound 6a

Compound 3 (l .OOg, 2.45 mmol) in N-methylimidazole (6.0 ml) was treated with iert-butyl 1-methylhydrazinecarboxylate (0.363 ml, 2.45 mmol) and the mixture was allowed to stir at 60 °C for 20 hours, cooled to room temperature and diluted with DMSO (30 mL) and acetic acid (10 mL). Direct purification using prepartive RP-MPLC provided intermediate compound 6a that was used without further purification. LCMS (M + H) = 537, 1H NMR (500 MHz, CDCI 3 ): δ 8.73 (s; 1 H); 7.34-7.29 (m, 1 H); 6.83-6.87 (m; 2 H); 4.44-4.48 (m; 3 H); 4.39 (dd; J = 10.8; 7.0 Hz; 1 H); 4.02 (s; 3 H); 3.41 (s; 3 H); 1.37-1.43 (m; 12 H).

Step B - Synthesis of Intermediate Compounds 6b-l and 6b-2

Compound 6a (1.03 g, 1.920 mmol) in ethanol (10 ml) treated at room

temperature with LiOH (2 M aqueous) (0.960 ml, 1.920 mmol). The mixture was allowed to stir at room temperature for 20 h and then neutralized with glacial acetic acid and concentrated to remove a majority of the ethanol. The resulting residue was purified using RP-MPLC to separately provide intermediate compound 6b- 1 and intermediate compound 6b -2. 6b- 1 : LCMS (M + H) = 409; 1H NMR (500 MHz, DMSO) δ 8.92 (s; 1 H); 7.50-7.55 (m; 1 H); 7.28 (td; J = 9.8; 2.5 Hz; 1 H); 7.11 (td; J = 8.5; 2.7 Hz; 2 H); 4.48 (s; 2 H); 3.82 (s; 3 H); 2.80 (s; 3 H); 6b-2: LCMS (M + H) = 509; 1H NMR (500 MHz, CDC1 3 ) δ 8.75 (s; 1 H); 7.31-7.34 (m; 1 H); 6.82-6.87 (m; 2 H); 4.47-4.48 (m; 2 H); 4.04 (s; 3 H); 3.46 (s; 3 H); 1.42 (s; 9 H).

Step C - Synthesis of Intermediate Compound 6c-l

Compound 6b-l (950 mg, 2.326 mmol) in DMF (4.0 ml) treated at 0 °C with PyClock (1549 mg, 2.79 mmol) and ethylamine (2 M in THF) (2.56 ml, 5.12 mmol). Stirred at room temperature for 6 hours. Quenched with water (1.0 mL). Neutralized with glacial acetic acid. Purification using preparative reverse-phase HPLC provided a solid after lyophilization. The solid was suspended in acetonitrile (50 mL) and gently heated until dissolved. Water (50 mL) was added. The mixture was aged in at -20 °C for 15 minutes. The resulting flocculant precipitate was filtered and washed with 50% aqueous acetonitrile (2 x 10 mL) to provide intermediate compound 6c- 1 as a solid. LCMS (M + H) = 436; 1H NMR (500 MHz, DMSO): δ 8.89 (s; l H); 8.67 (t; J = 5.6 Hz; 1 H); 7.51-7.56 (m; 1 H); 7.29 (td; J = 9.8; 2.6 Hz; 1 H); 7.11 (td; J = 8.5; 2.5 Hz; 1 H); 6.92-6.95 (m; 1 H); 4.48 (s; 2 H); 3.80 (s; 3 H); 3.25 (p; J = 6.7 Hz; 2 H); 2.79 (d; J = 5.8 Hz; 3 H); 1.10 (t; J = 7.2 Hz; 3 H).

Step C - Synthesis of Intermediate Compound 6c-2

Compound 6b-2 (2.654 g, 5.22 mmol) and PyClock (3.48 g, 6.26 mmol) in N,N- dimethylformamide (5 ml) was treated at room temperature with ethylamine (2 M in THF) (5.74 ml, 11.48 mmol). The misture was allowed to stir at room temperature for 16 h and then quenched by the addition of water (0.5 mL) and neutralized with glacial acetic acid. Purification using RP-MPLC (C18, 150g ISCO, 10 to 95% MeCN/water+0.1% TFA, 10CV) provided intermediate compound 6c-2. LCMS (M + H) = 536; 1H NMR (500 MHz, DMSO) δ 9.18 (s; 1 H); 8.78 (s; 1 H); 7.52-7.57 (m; 1 H); 7.27-7.31 (m; 1 H); 7.11 (td; J = 8.5; 2.5 Hz; 1 H); 4.50 (s; 2 H); 3.84 (s; 3 H); 3.32 (s; 3 H); 3.22 (t; J = 7.3 Hz; 2 H); 1.40 (br s; 9 H); 1.07 (t; J = 7.2 Hz; 3 H). Step D - Synthesis of Intermediate Compound 6d

Compound 6c- 1 (100 mg, 0.230 mmol) and ethyl 4-oxopiperidine-l-carboxylate (197 mg, 1.148 mmol) in dichloroacetic acid (1 mL) was heated at 100 °C for 8 hours, cooled to room temperature and diluted with acetonitrile. Direct purification using preparative reverse- phase HPLC provided intermediate compound 6d. LCMS (M + H) = 575, 1H NMR (500 MHz, CDCI3): δ 8.88 (s; 1 H); 7.28-7.33 (m; 1 H); 6.83-6.87 (m; 2 H); 4.50 (s; 2 H); 4.29 (br t; J = 18.32 Hz; 1 H); 4.15 (s; 3 H); 3.98 (br s; 1 H); 3.58-3.69 (m; 2 H); 3.10 (br s; 2 H); 2.87 (s; 3 H); 2.09-2.20 (m; 2 H); 1.73-1.79 (m; 1 H); 1.66 (d; J = 13.75 Hz; 1 H); 1.24-1.29 (m; 6 H). Step E - Synthesis of Compound 32

Compound 6d was dissolved in dioxane and treated with 1M aqueous LiOH at room temperature and then heated to 100 °C and allowed to stir at this temperature for 16 hours. The mixture was cooled to room temperature and neutralized with glacial acetic acid. Direct purification using preparative reverse-phase HPLC provided compound 32 as the TFA salt. LCMS (M + H) = 503, 1H NMR (500 MHz, CD 3 OD): δ 8.97 (s; 1 H); 7.43-7.48 (m; 1 H);

6.97-7.03 (m; 2 H); 4.49 (s; 2 H); 3.76-3.83 (m; 1 H); 3.67 (dq; J = 14.7; 7.2 Hz; 1 H); 3.40- 3.56 (m; 3 H); 3.30-3.31 (m; 3H); 2.92 (s; 3 H); 2.59-2.61 (m; 2 H); 2.15-2.17 (m; 2 H); 1.32 (t; J = 7.1 Hz; 3 H). The following compound of the present invention was made using the method described in the Example above and substituting the appropriate reactants and reagents.

Example 7

Preparation of Compound 34

Step A- Synthesis of Intermediate Compound 7a

A mixture of tert-butyl (3-(benzyloxy)-5-(5-(2,4-difluorobenzyl)-l,3,4- thiadiazol-2-yl)-2-(methylcarbamoyl)-4-oxopyridin-l(4H)-yl)( methyl)carbamate, prepared in a similar manner to the prior examples, (25 mg, 0.042 mmol) and (9H-fluoren-9-yl)methyl 4- oxoazepane-l-carboxylate (70.2 mg, 0.209 mmol) in dichloroacetic acid (0.25 ml) was heated at 100 °C for 4 hours. The mixture was cooled to room temperature and diluted with DMSO. Direct purification using preparative reverse-phase HPLC provided intermediate compound 7a. LCMS (M + H) = 725.

Step B- Synthesis of Compound 34

A solution of intermediate compound 7a (16 mg, 0.022 mmol) in DMF treated at room temperature with piperidine (0.5 mL) and stirred at room temperature for 20 hours. Direct purification using preparative reverse-phase HPLC provided compound 34 as its TFA salt. LCMS (M + H) = 503.

Example 8

Preparation of Compounds 35 and 36

Step A- Synthesis of Intermediate Compound 8a

A mixture of compound 1-2 (5000 mg, 10.32 mmol) and iert-butyl hydrazinecarboxylate (1364 mg, 10.32 mmol) in N-methylimidazole (20.0 ml) was heated at 60 °C for 14 hours. The mixture was cooled to room temperature, diluted with ethyl acetate (15 mL) and the organic phase was washed with 1 M aq citric acid (3 x 10 mL) and brine. The organic layer was dried over anhydrous sodium sulfate and filtered and the filtrate was concentrated in vacuo. Purification using column chromatography on silica gel (0 to 100% ethyl acetate/ heptane) provided intermediate compound 8a. LCMS (M + H) = 599, 1H NMR (500

MHz, DMSO): δ 8.90 (s; 1 H); 7.53-7.58 (m; 1 H); 7.27-7.37 (m; 6 H); 7.12 (td; J = 8.54; 2.57 Hz; l H); 5.19 (s; 2 H); 4.51 (s; 2 H); 4.26 (q; J = 7.1 Hz; 2 H); 1.42 (s; 9 H); 1.20 (t; J = 7.1 Hz; 3 H). Step B- Synthesis of Intermediate Compound 8b

Compound 8a (6.00 g, 10.02 mmol) in ethanol (20 ml) was treated at room temperature with 2 M aq LiOH (50.1 ml, 100 mmol). The mixture was heated at 75 °C for 48 hours, cooled to room temperature and concentrated in vacuo. The resulting residue was neutralized with glacial acetic acid in acetonitrile. Purification using preparative reverse-phase HPLC provided intermediate compound 8b as a powder . LCMS (M + H) = 571; 1H NMR (500 MHz, DMSO): δ 11.36 (br s; 1 H); 8.83 (s; 1 H); 7.53-7.58 (m; 1 H); 7.26-7.42 (m; 6 H); 7.12 (td; J = 8.5; 2.52 Hz; 1 H); 5.16 (s; 2 H); 4.51 (s; 2 H); 1.43 (s; 9 H).

Step C- Synthesis of Intermediate Compound 8c

A solution of compound 8b (1.00 g, 1.753 mmol) in DMF (5.0 ml) was treated at room temperature with PyClock (1.167 g, 2.103 mmol) and ethylamine (2 M in THF) (1.928 ml, 3.86 mmol). The mixture was allowed to stir at room temperature for 16 hours, quenched with water and neutralized with glacial acetic acid. Purification using RP-MPLC provided

intermediate compound 8c. LCMS (M + H) = 598; 1H NMR (500 MHz, DMSO): δ 11.30 (br s; 1 H); 8.77 (s; 1 H); 7.53-7.58 (m; 1 H); 7.28-7.42 (m; 6 H); 7.12 (td; J = 8.5; 2.6 Hz; 1 H); 5.17 (s; 2 H); 4.50 (s; 2 H); 3.19 (p; J = 6.7 Hz; 2 H); 1.44 (s; 9 H); 1.03 (t; J = 7.2 Hz; 3 H)

Step D- Synthesis of Intermediate Compound 8d

Compound 8c (50 mg, 0.084 mmol) and dihydrofuran-3(2H)-one (138 mg, 0.418 mmol) in acetic acid (1.0 ml) heated at 85 °C for 12 hours. The reaction was cooled to room temperature, then diluted with acetonitrile (4 mL). Purification of the resulting acetonitrile solution using preparative mass-guided reverse-phase HPLC provided intermediate compound 8d. LCMS (M + H) = 566. Step E- Synthesis of Intermediate Compound 8e and Intermediate Compound 8f

Compound 8d was dissolved in 5% aq DMF (1 mL) and treated with K2CO3 (5 equiv) and iodomethane (5 equiv). The mixture was allowed to stir at room temperature for 16 h and then quenched with the addition of glacial acetic acid. Purification using preparative reverse-phase HPLC provided a mixture of intermediate compounds 8e and 8f as the racemate. (1H NMR indicates confomational isomers at room temperature). LCMS (M + H) = 580; 1H

NMR (500 MHz, CDC13): δ 8.82 (d; J = 12.4 Hz; 1 H); 7.45-7.50 (m; 2 H); 7.26-7.29 (m; 3 H);

6.86-6.88 (m; 2 H); 5.62 (t; J = 10.5 Hz; 1 H); 5.30 (dd; J = 10.6; 3.28 Hz; 1 H); 4.52 (s; 2 H);

4.19 (d; J = 10.2 Hz; 1 H); 4.04-4.10 (m; 1 H); 3.86-3.95 (m; 2 H); 3.68 (d; J = 10.2 Hz; 1 H);

3.47 (d; J = 9.3 Hz; 1 H); 3.34 (td; J = 14.5; 7.2 Hz; 1 H); 3.11 (d; J = 9.6 Hz; 1H); 2.87 (s; 1 H); 2.77 (s; 2 H); 2.49 (t; J = 7.0 Hz; 1 H); 2.26 (d; J = 11.9 Hz; 1 H); 2.12 (t; J = 6.1 Hz; 1 H);

1.50-1.54 (m; 1 H); 1.28-1.33 (m; 3 H). Resolution to the enantiomers was accomplished by SFC-HPLC (Chiralcel OD, 21x250mm, 50% methanol+0.2%DEA, 55ml/min, 20mg/ml in methanol) to provide

intermediate compound 8e (enantiomer A, earlier eluting) and intermediate compound 8f (enantiomer B, later eluting).

Step F- Synthesis of Compound 35 and 36

A mixture of compound 8e (enantiomer A) (45 mg, 0.078 mmol) and Pd/C (15 mg, 0.141 mmol) in ethyl acetate (3 ml) was sparged with nitrogen. Hydrogen (1 atm) was introduced and the reaction mixture was allowed to stir at room temperature for 16 hours. After filtration and concentration of the filtrate, the resulting residue was purified using preparative reverse-phase HPLC to provide intermediate compound 8g. (1H NMR indicates confomational isomers at room temperature) LCMS (M + H) = 490; 1H NMR (500 MHz, DMSO): δ 8.71 (s; 1 H); 8.65 (s; 1 H); 7.50-7.55 (m; 2 H); 7.28 (td; J = 9.8; 2.5 Hz; 2 H); 7.11 (td; J = 8.5; 2.5 Hz; 2 H); 4.50 (s; 4 H); 4.32 (d; J = 10.8 Hz; 1 H); 3.99-4.05 (m; 4 H); 3.77-3.88 (m; 4 H); 3.64- 3.71 (m; 2 H); 2.86 (s; 3 H); 2.77 (s; 3 H); 2.59 (dt; J = 14.1; 6.9 Hz; 1 H); 2.35-2.46 (m; 2 H); 2.13 (dt; J = 13.3; 8.1 Hz; 1 H); 1.21-1.26 (m; 6 H).

Similar conditions to those used for the preparation of compound 35 provided compound 36 using intermediate compound 8f (enantiomer B).

The following compounds of the present invention were made using the method described in the Example above and substituting the appropriate reactants and reagents.

Exact

Compound Structure Mass

ΓΜ+Η1+

37 504

OH O

38 458

OH O Example 8

Preparation of Compound 39

Step A- Synthesis of Intermediate Compound 8a

To a solution of 3-(aminomethyl) tetrahydrofuran-3 -amine dihydrochloride (2.0 g, 10.58 mmol) and triethylamine (2.95 mL, 21.15 mmol) in methanol (35 mL) was added di-tert- butyl dicarbonate (1.38 g, 6.35 mmol) at 0 °C, and the mixture was allowed to stir at room temperature for 6 hours. The mixture was concentrated to remove the methanol, diluted with water (100 mL) and extracted with dichloromethane (8x 80 mL). The combined organic portion was dried over anhydrous Na 2 S0 4 and concentrated to provide intermediate compound 8 a as an oil. 1H NMR (400 MHz, CDC1 3 ) δ 5.07 (s, 1H), 3.89-4.00 (m, 3H), 3.62 (d, J = 8.8 Hz, 1H), 3.49 (d, J = 8.4 Hz, 1H), 3.21-3.25 (m, 2H), 1.95-1.98 (m, 1H), 1.72-1.75 (m, 1H), 1.45 (s, 9H).

Step B- Synthesis of Intermediate Compound 8b

To a solution of intermediate compound 8a (1.35 g, 6.24 mmol), 3-(benzyloxy)-4- oxo-4H-pyran-2-carboxylic acid (1.69 g, 6.87 mmol) and triethylamine (1.914 mL, 13.73 mmol) in DMF (25 mL) was added HATU (2.6 g, 6.87 mmol). The mixture was allowed to stir at room temperature for 1.5 hours. The mixture was concentrated to remove the DMF, then diluted with water (100 mL), extracted with ethyl acetate (4 x 100 mL). The combined organic portion was washed with brine (200 mL), dried over anhydrous Na 2 S0 4 and concentrated in vacuo. The resulting residue was purified using column chromatography on silica gel (petroleum ether: ethyl acetate, 10: 1 to 1.5: 1) to provide intermediate compound 8b as an oil. LRMS (+ESI) m/z 445.5. 1H NMR (400 MHz, CD 3 OD) δ 8.06-8.08 (m, 1H), 7.37-7.53 (m, 5H), 6.51-6.54 (m, 1H), 5.31- 5.38 (m, 2H), 3.75-3.80 (m, 3H), 3.64-3.67 (m, 1H), 3.47-3.49 (m, 1H), 3.38-3.41 (m, 1H), 1.96- 1.99 (m, 1H), 1.81-1.83 (m, 1H), 1.40 (s, 9H).

Step C- Synthesis of Intermediate Compound 8c

A mixture of intermediate compound 8b (2.0 g, 4.50 mmol) in TFA (2

mL)/dichloromethane (10 mL) was allowed to stir at room temperature for 3 hours. The mixture was concentrated and the resulting residue was diluted with EtOH (50 mL). The mixture was heated at 100 °C for 16 hours, cooled to room temperature and concentrated in vacuo. The resulting residue was purified using column chromatography on silica gel (petroleum ether: ethyl acetate 1 : 1 to dichloromethane: methanol 20: 1) to provide intermediate compound 8c as an oil. 1H NMR (400 MHz, CD 3 OD) δ 8.09-8.11 (m, 1H), 7.29-7.45 (m, 5H), 6.57-6.61 (m, 1H), 5.17- 5.28 (m, 1H), 3.54-4.21 (m, 7H), 1.82-2.44 (m, 2H).

Step D- Synthesis of Intermediate Compound 8d

A mixture solution of intermediate compound 8c (500 mg, 1.532 mmol) and N- bromosuccinimide (409 mg, 2.298 mmol) in THF (8 mL) was allowed to stir at room

temperature for 4 hours. The mixture was concentrated and the resulting residue was purified using preparative TLC (dichloromethane: methanol, 10: 1) to provide intermediate compound 8d as an oil. 1H NMR (400 MHz, CD 3 OD) δ 8.25 (s, 1H), 7.43-7.45 (m, 2H), 7.30-7.32 (m, 3H), 5.28-5.29 (m, 2H), 4.22 (s, 2H), 3.90-3.94 (m, 2H), 3.56 (d, J = 8.8 Hz, 1H), 3.44 (d, J = 9.2 Hz, 1H), 1.97-2.03 (m, 1H), 1.83-1.87 (m, 1H).

Step E- Synthesis of Intermediate Compound 8e

A mixture solution of intermediate compound 8d (80 mg, 0.20 mmol) and iodomethane (34 mg, 0.24 mmol) in DMF (4 mL) was allowed to stir at 80 °C for 2.5 hours. The mixture was diluted with water (30 mL), extracted with ethyl acetate (3x 25 mL). The combined organic extracts was washed with brine (30 mL) and concentrated to provide the residue, which was purified using preparative TLC (dichloromethane : methanol, 10: 1) separation to provide intermediate compound 8e as a solid. LRMS (+ESI) m/z: 420.9. 1H NMR (400 MHz, CD 3 OD) δ 8.22 (s, 1H), 7.40-7.43 (m, 2H), 7.31-7.33 (m, 3H), 5.22-5.30 (m, 2H), 4.21-4.22 (m, 2H), 3.79- 3.96 (m, 3H), 3.43 (d, J = 10.4 Hz, 1H), 3.06 (s, 3H), 1.91-1.94 (m, 2H).

Step F- Synthesis of Intermediate Compound 8f A solution of intermediate compound 8e, N,N-diisopropylethylamine (0.056 mL, 0.322 mmol) and (2,4-difluorophenyl)methanamine (46.1 mg, 0.322 mmol) in DMSO (3 mL) was treated at room temperature with Pd(PPh 3 ) 4 (9.3 mg, 8.1 μιηοΐ) . The mixture was heated at 90 °C under carbon monoxide (1 atm) for 16 hours. The mixture was cooled to room

temperature, diluted with water (30 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic extracts was washed with brine (30 mL), dried over anhydrous Na 2 S0 4 and concentrated in vacuo. The resulting residue was purified using preparative TLC

(dichloromethane: methanol, 10: 1) to provide intermediate compound 8f as an oil. LRMS (+ESI) m/z: 510.2. 1H NMR (400 MHz, CD 3 OD) δ 8.50 (s, 1H), 7.40-7.42 (m, 2H), 7.30-7.32 (m, 3H), 6.97-7.02 (m, 3H), 5.23-5.30 (m, 2H), 4.63-4.66 (m, 2H), 4.32 (s, 2H), 3.82-3.98 (m, 3H), 3.45 (d, J = 10.4 Hz, 1H), 3.07 (s, 3H), 1.921-1.96 (m, 2H).

Step G- Synthesis of Compound 39

A mixture of intermediate compound 8f (10 mg, 0.02 mmol) and lithium chloride (25.2 mg, 0.595 mmol) in DMF (3 mL) was allowed to stir at 90 °C for 5 hours. The mixture was filtered and the filtrate was purified using preparative HPLC to provide compound 39 as a solid. LRMS (+ESI) m/z: 420.0. 1H NMR (400 MHz, CD 3 OD) δ 8.47 (s, 1H), 7.41-7.51 (m, 1H), 6.93-7.01 (m, 2H), 4.64 (s, 2H), 4.40-4.43 (m, 2H), 4.08-4.11 (m, 2H), 3.90-3.94 (m, 1H), 3.66 (d, J = 10.8 Hz, 1H), 3.20 (s, 3H), 2.05-2.31 (m, 2H).

Example 9

Preparation of Compound 40

Step A- Synthesis of Intermediate Compound 9a

A solution of 4-(aminomethyl)tetrahydro-2H-pyran-4-amine hydrochloride (450 mg, 2.70 mmol) in THF (40 mL) was added triethylamine (1.505 mL, 10.80 mmol), Boc 2 0 (0.627 mL, 2.70 mmol) at room temperature. The mixture was allowed to stir for 12 h at room temperature and then water (40 mL) was added. The mixture was extracted with

dichloromethane (5x 30 mL) and the combined organic portion was washed with water (2x 30 mL), dried over anhydrous Na 2 S0 4 and concentrated to provide intermediate compound 9a as a colorless oil. LRMS (+ESI) m/z: 231.1. 1H NMR: (400 MHz, CDC1 3 ) δ 4.96 (br, 1H), 3.73-3.76 (m, 4H), 3.10 (d, J = 6.0 Hz, 2H), 1.671-1.67 (m, 2H), 1.45 (s, 9H), 1.34-1.38 (m, 2H).

Step B- Synthesis of Intermediate Compound 9b

To a solution of intermediate compound 9a (1 g, 4.34 mmol) in DMF (20 mL) was added, at room temperature, triethylamine (0.822 g, 8.12 mmol), HATU (1.699 g, 4.47 mmol) and 3-(benzyloxy)-4-oxo-4H-pyran-2-carboxylic acid (0.982 g, 4.26 mmol). The mixture was allowed to stir for 12 h at room temperature and then treated with water (30 mL). The mixture was extracted with ethyl acetate (5x 15 mL) and the combined organic portion was washed with water (2x 25 mL) and brine (25 mL), dried over anhydrous Na 2 S0 4 and concentrated, the resulting residue was purified using column chromatography on silica gel (petroleum ether: ethyl acetate = 1 : 1 to 1 : 10) to provide intermediate compound 9b as a solid.

Step C- Synthesis of Intermediate Compound 9c

To a solution of intermediate compound 9b (1.5 g, 3.27 mmol) in CH 2 CI 2 (35 mL) was added TFA (7 mL). The mixture was allowed to stir at room temperature for 2 h and then concentrated to provide intermediate compound 9c as an oil. LRMS (+ESI) m/z: 359.0. 1 FiNMR: (300 MHz, CD 3 OD) δ 8.13 (d, J = 5.4 Hz, 1H), 7 ' .52-7 ' .55 (m, 2H), 7.41-7.44 (m, 3H), 6.60 (d, J = 5.4 Hz, 1H), 5.44 (s, 2H), 3.34-3.85 (m, 6H), 1.58-1.90 (m, 4H).

Step D- Synthesis of Intermediate Compound 9d

A solution of intermediate compound 9c (0.9 g, 1.976 mmol)) in EtOH (15 mL) was heated at reflux for 12 h and then concentrated and purified using preparative HPLC to provide intermediate compound 9d as a solid. LRMS (+ESI) m/z: 359.0. 1H NMR: (400 MHz, CD3OD) δ 7.67 (d, J = 7.6 Hz, 1H), 7.37-7.40 (m, 2H), 7.26-7.29 (m, 3H), 6.53 (d, J = 7.6 Hz, 1H), 5.30 (s, 2H), 4.11 (s, 2H), 3.62-3.71 (m, 4H), 1.52-1.59 (m, 2H), 1.29-1.32 (m, 2H).

Step E- Synthesis of Intermediate Compound 9e

To a solution of intermediate compound 9d (30 mg, 0.088 mmol) in dichloromethane (4 mL) was added N-bromosuccinimide (31.4 mg, 0.176 mmol). The mixture was allowed to stir at room temperature for 2 hours. The mixture was washed with water (2 mL)and brine (2 mL) and the comgined organic portion was concentrated and purified using preparative TLC (dichloromethane: methanol= 10: 1) to provide intermediate compound 9e as an oil. LRMS (+ESI) m/z: 420.7. 1H NMR: (400 MHz, CD 3 OD) δ 8.23 (s, 1H), 7.37-7.40 (m, 2H), 7.27-7.29 (m, 3H), 5.32 (s, 2H), 4.13 (s, 2H), 3.63-3.71 (m, 4H), 1.54-1.61 (m, 2H), 1.31-1.34 (m, 2H).

Step F- Synthesis of Intermediate Compound 9f

To a solution of intermediate compound 9e (0.02 g, 0.048 mmol) in DMF (2 mL) was added Cs 2 C0 3 (0.062 g, 0.19 mmol) and CH 3 I (0.068 g, 0.48 mmol), then the mixture was allowed to stir at 80 °C for 3 hours, then water (4 mL) was added, extracted with

dichloromethane (2 mL x6), the combined organic portion was dried over anhydrous Na 2 S0 4 , filtered and the filtrate was concentrated in vacuo. The resulting residue was purified using preparative TLC (dichloromethane: methanol, 10:1) to provide intermediate compound 9f as an oil. LRMS (+ESI) m/z: 434.8. 1 HNMR: (400 MHz, CD 3 OD) δ 8.30 (s, 1H), 7.36-7.38 (m, 2H), 7.27-7.29 (m, 3H), 5.24 (s, 2H), 4.39 (s, 2H), 3.78-3.82 (m, 2H), 3.51-3.57 (m, 2H), 3.01 (s, 3H), 1.93-2.01 (m, 2H), 1.19-1.23 (m, 2H).

Step G- Synthesis of Intermediate Compound 9g

A solution of intermediate compound 9f (10 mg, 0.023 mmol) in DMSO (2 mL) was treated with (2,4-difluorophenyl)methanamine (33.03 mg, 0.231 mmol), N,N- diisopropylethylamine (14.91 mg, 0.115 mmol) and Pd(PPh 3 ) 4 (2.67 mg, 0.0023 mmol) at room temperature under an atmosphere of carbon monoxide. The mixture was allowed to stir at 90 °C under carbon monoxide (1 atm) for 15 h and then diluted with water (4 mL). The mixture was extracted with ethyl acetate (3 mL x 4) and the combined organic portion was washed with water (3 mL x2) and concentrated in vacuo. The resulting residue was purified using preparative TLC (dichloromethane : methanol, 20: 1) to provide intermediate compound 9g as a solid. LRMS (+ESI) m/z: 523.9. 1 HNMR: (400 MHz, CD 3 OD) δ 8.57 (s, 1H), 7.30-7.57 (m, 4H), 6.97-7.12 (m, 4H), 5.27 (s, 2H), 4.52 (s, 2H), 4.19 (s, 2H), 3.81-3.83 (m, 2H), 3.58-3.64 (m, 2H), 3.05 (s, 3H), 1.28-1.30 (m, 4H).

Step H- Synthesis of Compound 40

To a solution of intermediate compound 9g (5 mg, 0.010 mmol) in DMF (2 mL) was added LiCl (12.15 mg, 0.287 mmol) at room temperature under N 2 atmosphere, then the mixture was allowed to stir at 95 °C for 2 hours, the mixture was purified using preparative reverse-phase HPLC to provide compound 40 as a solid. LRMS (+ESI) m/z: 434. 1H NMR: (400 MHz, CD 3 OD) δ 8.53-8.69 (m, 1H), 7.40-7.43 (m, 1H), 3.77-6.93 (m, 2H), 4.64 (s, 2H), 3.89-3.90 (m, 2H), 3.70-3.71 (m, 2H), 3.16 (s, 3H), 2.24-2.25 (m, 2H), 1.59-1.63 (m, 2H), 1.27- 1.28 (m, 2H).

Example 10

Preparation of Compound 41

10b 41

Step A- Synthesis of Intermediate Compound 10a

A mixture of 8d (30 mg, 0.074 mmol), Cs 2 C0 3 (48.2 mg, 0.148 mmol) and 1- iodopropane (25.2 mg, 0.148 mmol) in DMF (3 mL) was allowed to stir at room temperature for 3 hours. The mixture was diluted with water (30 mL) and extracted with ethyl acetate (4x 20 mL) The combined organic portion was dried over anhydrous Na 2 S0 4 , filtered and the filtrate was concentrated in vacuo. The resulting residue was purified using preparative TLC

(dichloromethane: methanol, 10: 1) to provide intermediate compound 10a as an oil. LRMS (+ESI) m/z: 448.9. 1H NMR (400 MHz, CD 3 OD) δ 8.23 (s, 1H), 7.31-7.39 (m, 5H), 5.28 (s, 2H), 4.19-4.20 (m, 2H), 3.86-3.96 (m, 2H), 3.65-3.67 (m, 1H), 3.40-3.41 (m, 3H), 1.93 (brs, 2H), 1.65-1.69 (m, 2H), 0.96-1.00 (m, 3H).

Step B- Synthesis of Intermediate Compound 10b

A mixture of intermediate compound 10a (18 mg, 0.040 mmol), N,N-diisopropyl- ethylamine (0.056 mL, 0.322 mmol) and (2,4-difluorophenyl)methanamine (46.1 mg, 0.322 mmol) in DMSO (3 mL) was treated with Pd(Ph 3 P) 4 (9.30 mg, 8.05 μιηοΐ). The solution was heated at 90 °C under carbon monoxide (1 atm) for 16 hours. The mixture was diluted with water (30 mL) and extracted with ethyl acetate (3x 20 mL). The combined organic portion was washed with brine (30 mL), dried over anhydrous Na 2 S0 4 , filtered and the filtrate was

concentrated The resulting residue was purified using preparative TLC (dichloromethane :

methanol, 8: 1) to provide intermediate compound 10b as an oil. LRMS (+ESI) m/z: 538.3. 1H NMR (400 MHz, CD 3 OD) δ 8.48 (s, 1H), 7.39-7.47 (m, 3H), 7.30-7.31 (m, 3H), 6.97-6.99 (m, 2H), 5.28-5.31 (m, 2H), 4.66 (s, 2H), 4.29-4.31 (m, 2H), 3.89-3.98 (m, 2H), 3.67-3.69 (m, 1H), 3.39-3.45 (m, 3H), 1.93-1.97 (m, 2H), 1.64-1.72 (m, 2H), 0.96-1.00 (m, 3H).

Step C- Synthesis of Compound 41 A mixture of intermediate compound 10b (8 mg, 0.015 mmol) and lithium chloride (25.2 mg, 0.595 mmol) in DMF (3 mL) was heated at 90 °C for 5 hours. The mixture was cooled to room temperature and filtered. The filtrate was purified using preparative reverse- phase HPLC to provide compound 41 as a solid. LRMS (+ESI) m/z: 448.2. 1H NMR (400 MHz, DMSO-de) δ 12.4 (brs, 1H), 10.40-10.42 (m, 1H), 8.45 (s, 1H), 7.40-7.41 (m, 1H), 7.21-7.27 (m, 1H), 7.07-7.08 (m, 1H), 4.54-4.55 (m, 2H), 4.42-4.43 (m, 2H), 4.01-4.03 (m, 1H), 3.80-3.87 (m, 2H), 3.59-3.61 (m, 1H), 3.36-3.41 (m, 2H), 2.08-2.12 (m, 2H), 1.62-1.65 (m, 2H), 0.89-0.93 (m, 3H). The following compounds of the present invention were made using the method described in the Example above and substituting the appropriate reactants and reagents.

Example 11

Preparation of Compound 44

Step A- Synthesis of Intermediate Compound 11a

To a solution of intermediate compound 9e (20 mg, 0.048 mmol) in DMF (2 mL) was added NaH (2.86 mg, 0.072 mmol) at 5 °C. The mixture was allowed to stir for 0.5 hours, then 1-iodopropane (16.22 mg, 0.095 mmol) was added and the mixture was allowed to stir at 5 °C for 3 hours. Water (4 mL) was added dropwise and the resulting mixture was extracted with ethyl acetate (4 mL x 5). The combined organic portion was washed with water (4 mLx2) and brine (4 mL), dried over anhydrous Na 2 S0 4 , filtered and the filtrate was concentrated in vacuo. The resulting residue was purified using preparative TLC (ethyl acetate) to provide intermediate compound 1 la as an oil. LRMS (+ESI) m/z. 463; 1H NMR (400 MHz, CD 3 OD) δ 8.30 (s, 1H), 7.38-7.35 (m, 2H), 7.29-7.27 (m, 3H), 5.26 (s, 2H), 4.40 (s, 2H), 3.82-3.78 (m, 2H), 3.53 (t, J = 11.6 Hz, 2H), 3.41 (t, J = 8.0 Hz, 2H), 1.98-1.91 (m, 2H), 1.64-1.59 (m, 2H), 1.26- 1.22 (m, 2H), 0.97 (t, J = 7.6 Hz, 3H).

Step B- Synthesis of Intermediate Compound lib

To a solution of intermediate compound 11a (15 mg, 0.033 mmol) in DMSO (2 mL) was added N,N-diisopropylethylamine (21.01 mg, 0.163 mmol), (2,4- difluorophenyl)methanamine (46.5 mg, 0.325 mmol) and Pd(PPh 3 ) 4 (3.76 mg, 3.25 μιηοΐ) at room temperature. The mixture was heated at 90 °C under carbon monoxide (1 atm) for 15 h and then diluted with water (6 mL) . The mixture was extracted with ethyl acetate (4 mL x 4) and the combined organic portion was washed with water (4 mL x2) and brine (4 mL), dried over anhydrous Na 2 S0 4 , filtered and the filtrate was concentrated in vacuo. The resulting residue was purified using preparative TLC (ethyl acetate) to provide intermediate compound l ib as an oil. LRMS (+ESI) m/z: 552.2. 1 HNMR: (400 MHz, CD 3 OD) δ 8.57 (s, 1H), 7.56-7.55 (m, 1H), 7.48-7.46 (m, 1H), 7.39-7.38 (m, 2H), 7.30-7.29 (m, 3H), 7.12-7.09 (m, 1H), 5.28 (s, 2H), 4.66-4.65 (m, 2H), 4.52 (s, 2H), 3.83-3.79 (m, 2H), 3.62-3.59 (m, 2H), 3.44 (t, / = 8.0Hz, 2H), 2.01-1.94 (m, 2H), 1.65-1.61 (m, 2H), 1.24-1.22 (m, 2H), 0.98 (t, / = 7.2Hz, 3H).

Step C- Synthesis of Compound 44

To a solution of intermediate compound l ib (10 mg, 0.018 mmol) in DMF (2 mL) was added lithium chloride (23.06 mg, 0.544 mmol) at room temperature. The mixture was heated at 90 °C for 3 hours, cooled to room temperature and directly purified using preparative reverse-phase HPLC to provide compound 44. LRMS (+ESI) m/z: 462.2. 1H NMR: (400 MHz, DMSO- ) δ 12.51 (s, 1H), 10.44 (t, / = 6.0 Hz, 1H), 8.63 (s, 1H), 7.42-7.38 (m, 1H), 7.24 (t, / = 9.6 Hz, 1H), 7.07 (t, / = 7.2 Hz, 1H), 4.68 (s, 2H), 4.55 (d, / = 5.6 Hz, 2H), 3.78-3.75 (m, 2H), 3.68-3.65 (m, 2H), 3.48 (t, / = 8.0 Hz, 2H), 2.08-2.03 (m, 2H), 1.60-1.57 (m, 4H), 0.92 (t, / = 7.2Hz, 3H).

The following compound of the present invention was made using the method described in the Example above and substituting the appropriate reactants and reagents.

Example 12

Preparation of Compound 46

Step A- Synthesis of Intermediate Compound 12a

To a solution of compound 3-(benzyloxy)-4-oxo-6-(((tetrahydro-2H-pyran-2-yl) oxy)methyl)-4H-pyran-2-carboxylic acid (233 mg, 0.648 mmol), intermidiate compound 8a (140 mg, 0.648 mmol) and N,N-diisopropylethylamine (167 mg, 1.296 mmol) in DMF (10 mL) was added HATU (492 mg, 1.296 mmol) and HO AT (176 mg, 1.296 mmol). The mixture was allowed to stir at room temperature for 16 h and then diluted with water (20 mL) and extracted with ethyl acetate (4x 20 mL). The combined organic portion was washed with brine (20 mL), dried over anhydrous Na 2 S0 4 , filtered and the filtrate was concentrated in vacuo. The resulting residue was purified using preparative TLC (petroleum ether: ethyl acetate, 5: 1) to provide intermediate compound 12a as an oil. LRMS (+ESI) m/z 559.2.

Step B- Synthesis of Intermediate Compound 12b

To a solution of intermediate compound 12a (160 mg, 0.287 mmol) in ethyl acetate (10 mL) was added a solution of HCl in ethyl acetate (4 M, 4 mL) at room temperature. The mixture was allowed to stir for 2 h at room temperature and then concentrated in vacuo to provide intermediate compound 12b, which used without further purification. LRMS (+ESI) m/z 375.1.

Step C- Synthesis of Intermediate Compound 12c

Intermediate compound 12b (150 mg, 0.401 mmol) in EtOH (50 mL) was heated at 100 °C for 54 hours. The mixture was concentrated and the resulting residue was purified using preparative TLC (PE: EA = 2: 1) to provide intermediate compound 12c as an oil. LRMS (+ESI) m z 357.1.

Step D- Synthesis of Intermediate Compound 12d

A mixture of intermediate compound 12c (20 mg, 0.056 mmol) in DMF (3 mL) was treated at 0 °C with NaH (10 mg, 0.25 mmol) and iodomethane (31 mg, 0.224 mmol) and the mixture was allowed to stir at 0 °C for 2 hours. The mixture was diluted with water (30 mL) and extracted with ethyl acetate (3x 25 mL). The combined organic portion was washed with brine (30 mL) and concentrated in vacuo. The resulting residue was purified using preparative TLC to provide intermediate compound 12d as a solid. LRMS (+ESI) m/z 385.1.

Step E- Synthesis of Intermediate Compound 12e

A solution of intermediate compound 12d (15 mg, 0.039 mmol) in CH3CN (5 mL) was treated with N-bromosuccinimide (10 mg, 0.058 mmol), stirred at room temperature for 1 h and then concentrated in vacuo. The resulting residue was purified using preparative TLC to provide intermediate compound 12e. LRMS (+ESI) m/z 463.1.

Step F- Synthesis of Intermediate Compound 12f

A solution of 9'-(benzyloxy)-7'-bromo-6'-(methoxymethyl)-2'-methyl-4,5- dihydro-2H-spiro[furan-3,3'-pyrido[l,2-a]pyrazine]- ,8'(2'H,4'H)-dione (20 mg, 0.041 mmol) in DMSO (1 mL) and methanol (3 mL) was treated with (2,4-difluorophenyl)methanamine (60 mg, 0.419 mmol), N,N-diisopropylethylamine (60 mg, 0.465 mmol) and Pd(PPh 3 ) 4 (10 mg, 0.0086 mmol) at room temperature . The mixture was heated at 80 °C under carbon monoxide (1 atm) for 15 hours, cooled to room temperature and diluted with water (4 mL). The mixture was extracted with ethyl acetate (4x 3 mL) and the combined organic portion was washed with water (2 x 3 mL), concentrated and the resulting residue was purified using preparative TLC

(dichloromethane: methanol = 20: 1) to provide intermediate compound 12f as a solid. LRMS (+ESI) m/z 554.2. Step G- Synthesis of Compound 46

A solution of intermediate compound 12f (20 mg, 0.035 mmol) in dichloromethane (1 mL) was treated with TFA (3 mL) at 25 °C. The reaction mixture was allowed to stir at 25 °C for 2 h and then conentrated. The resulting residue was purified using preparative reverse-phase HPLC to provide compound 46. LRMS (+ESI) m/z =: 464.1. 1H NMR (400 MHz, MeOD) δ 7.85-7.90 (m, 1H), 6.96-7.01(m, 2H), 4.32-4.60 (m, 6H), 4.04-4.13 (m, 2H), 3.89-3.93 (m, 1H), 3.70-3.72 (m, 1H), 3.34 (s, 3H), 3.20 (s, 3H), 2.20-2.33 (m, 2H).

Example 13

Preparation of Compound 47

Step A- Synthesis of Intermediate Compound 13a and Intermediate Compound 13b

3-(aminomethyl)tetrahydrofuran-3-amine dihydrochloride salt (10.0 g, 52.9 mmol) and trityl chloride (16.2 g, 58.2 mmol) in dichloromethane (100 ml) was cooled to 0 °C and treated with triethylamine (24.33 ml, 175 mmol). The ice bath was removed and the mixture was allowed to stir at room temperature for 48 hours. The reaction mixture was filtered and the filtrated was directly loaded onto a dry pre-packed silica column. Purification using column chromatography on silica gel (0 to 100% Ethyl acetate/ hexanes) provided 3- ((tritylamino)methyl)tetrahydrofuran-3-amine. LCMS (M + H) = 359; 1H NMR (500 MHz,

CDCI 3 ): δ 7.50 (d; J = 7.8 Hz; 6 H); 7.39 (d; J = 7.9 Hz; 6 H); 7.18 (t; J = 7.3 Hz; 3 H); 4.01 (q; J = 8.0 Hz; 1 H); 3.79 (bs; 1 H); 3.68 (d; J = 10.0 Hz; 1 H); 3.57 (d; J = 9.2 Hz; 1 H); 2.31 (s; 2 H); 1.83 (s; 2 H). Resolution to the enantiomers was accomplished with SFC (OJ-H column, 40% 2-propanol + diethylamine/ C0 2 ) to provide 13a as enantiomer A (earlier eluting) and 13b as enantiomer B (later eluting). Step B- Synthesis of Intermediate Compound 13c

Compound 13a (1.70 g, 4.74 mmol) in dichloromethane (20.0 ml) cooled to 0 °C and treated with triethylamine (1.322 ml, 9.48 mmol) and methoxyacetyl chloride (0.477 ml, 5.22 mmol). Stirred at 0 °C for 1 hour. Directly loaded onto a 120 g silica column with hexanes. Purification using column chromatography on silica gel (0 to 100% EtOAc/ heptane; eluted at 90% EtOAc) provided intermediate compound 13c as a wax. LCMS (M + H) = 431; l U NMR (500 MHz, CDCI 3 ): δ 7.50-7.40 (m; 6 H); 7.32-7.23 (m; 6 H); 7.15-7.23 (m; 3 H); 6.97 (s; 1 H); 3.92-4.00 (m; 3 H); 3.83 (s; 3 H); 3.72 (d; J = 9.5 Hz; 1 H); 3.45 (s; 3 H); 2.56-2.49 (m; 2 H); 2.35 (bs, 1H) 1.98-1.86 (m; 1 H). Step C- Synthesis of Intermediate Compound 13d

Intermediate compound 13c (1.614 g, 3.75 mmol) in tetrahydrofuran (20 ml) was cooled to 0 °C and treated with a slurry of lithium aluminum hydride (0.300 g, 7.90 mmol) in THF (2 x 5 ml). The resulting mixture was heated under nitrogen at reflux for 4 hours, cooled to 0 °C and sequentially treated with water (0.30 mL), 15% w/w aqueous NaOH (0.30 mL), and water (0.90 mL). The resulting mixture was filtered through a pad of Solka-Floc and the filtrate was concentrated in vacuo. The resulting residue was purified using column chromatography on silica gel (0 to 100% EtOAc/ heptane) to provide intermediate compound 13d. LCMS (M + H) = 417. Step D- Synthesis of Intermediate Compound 13e

3-(benzyloxy)-4-oxo-4H-pyran-2-carboxylic acid (520 mg, 2.11 mmol) in THF (6 mL) treated at room temperature with l-(chloro-l-pyrrolidinylmethylene)pyrrolidinium hexafluorophosphate (843 mg, 2.54 mmol) and N,N-diisopropylethylamine (0.738 mL, 4.23 mmol), stirred at room temperature for 10 minutes and then treated with a solution of

intermediate compound 13d (880 mg, 2.113 mmol) in THF (2x2 mL) at room temperature. The mixture was allowed to stir at room temperature for 5 hours. The crude reaction mixture was directly loaded onto a dry silica gel column. Purification using column chromatography on silica gel (0 to 100% ethyl acetate/ hexanes) provided intermediate compound 13e. LCMS (M + H) = 645; Step E- Synthesis of Intermediate Compound 13f

Intermediate compound 13e (1.30 g, 2.02 mmol) in CH 2 CI 2 (20 ml) was treated dropwise with TFA (1.55 ml, 20.16 mmol) and the mixture was allowed to stir at room temperature for 16 hours. After concentration, the resulting residue was dissolved in N- methylimidazole (10 ml) heated at 80 °C for 16 hours. The mixture was cooled to room temperature and neutralized with glacial acetic acid. Purification using preparative reverse-phase HPLC provided intermediate compound 13f. LCMS (M + H) = 385. Step F- Synthesis of Intermediate Compound 13g

Intermediate compound 13f (180.8 mg, 0.470 mmol) in CH 2 C1 2 (5 mL) was treated with N-bromosuccinimide (92 mg, 0.517 mmol) at room temperature. The reaction mixture was allowed to stir for 2 h at room temperature. After concentration, the resulting residue was purified using column chromatography on silica gel (0 to 10% methanol/ CH 2 C1 2 / ) to provide intermediate compound 13g. LCMS (M + H) = 463.

Step G- Synthesis of Intermediate Compound 13h

A mixture of intermediate compound 13g (99 mg, 0.214 mmol), N,N- diisopropylethylamine (0.112 mL, 0.641 mmol), 2,4-difluorobenzylamine (0.058 mL, 0.470 mmol) and tetrakis(triphenylphosphine) palladium (123 mg, 0.107 mmol) in DMSO (2 mL) was sparged with nitrogen for 10 minutes and then carbon monoxide for 10 minutes. The reaction solution was heated at 90 °C under carbon monoxide (1 atm) for 8 hours. The mixture was cooled to room temperature, filtered and the filtrate was purified using preparative reverse-phase HPLC to provide intermediate compound 13h. LCMS (M + H) = 554.

Step H- Synthesis of Compound 47

Intermediate compound 13h (132 mg, 0.238 mmol) in TFA (1 mL, 12.98 mmol) was allowed to stir at room temperature for 3 hours. The mixture was diluted with aqueous DMSO and purified using preparative reverse-phase HPLC to provide compound 47. LCMS (M + H) = 464; 1H NMR (500 MHz, DMSO): δ 12.33 (s; 1 H); 10.39 (t; J = 6.0 Hz; 1 H); 8.43 (s; 1 H); 7.36-7.41 (m; 1 H); 7.22-7.26 (m; 1 H); 7.04-7.08 (m; 1 H); 4.53 (d; J = 5.9 Hz; 2 H); 4.46 (d; J = 13.3 Hz; 1 H); 4.35 (d; J = 13.3 Hz; 1 H); 4.06 (d; J = 10.3 Hz; 1 H); 3.98 (td; J = 8.6; 4.7 Hz; 1 H); 3.66-3.76 (m; 2 H); 3.51-3.63 (m; 4 H); 3.27 (s, 3H) 2.06-2.21 (m; 2 H). Similar conditions to those used for the preparation of compound 13 provided compound 14 using intermediate 13b (enantiomer B, later eluting).

The following compound of the present invention was made using the method described in the Example above and substituting the appropriate reactants and reagents.

Example 14

Preparation of Compound 49

49

Step A- Synthesis of Intermediate Compound 14a

A mixture of l-(2,4-difluorobenzyl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxabor olan-2- yl)-lH-pyrazole (26.7 mg, 0.083 mmol) and 9'-(benzyloxy)-7'-bromo-2'-methyl-4,5-dihydro-2H- spiro [furan-3,3'-pyrido[l,2-a]pyrazine]- ,8' (2'H,4'H)-dione (30 mg, 0.07 mmol) and Cs 2 C0 3 (27.2 mg, 0.083 mmol) in water (0.1 mL) and dioxane (1 mL) was treated with Pd(Ph 3 P) 4 (7.23 mg, 6.26 μιηοΐ) . The mixture was heated under microwave heating at 130 °C for 1 hour. The mixture was directly purified using preparative TLC (dichloromethane: methanol, 8: 1) to provide intermediate compound 14a as a solid. LRMS (+ESI) m/z. 533.0. 1H NMR (400 MHz, CD 3 OD) δ 8.53 (s, 1H), 8.22 (s, 1H), 8.06 (s, 1H), 7.27-7.38 (m, 6H), 6.97-6.99 (m, 2H), 5.42 (s, 2H), 5.23-5.24 (m, 2H), 4.23 (s, 2H), 3.75-3.94 (m, 3H), 3.40-3.43 (m, 1H), 3.04 (s, 3H), 1.92-1.93 (m, 2H).

Step B- Synthesis of Compound 49

A mixture of intermediate compound 14a and lithium chloride (25.2 mg, 0.595 mmol) in DMF (3 mL) was heated at 90°C for 5 hours. The mixture was filtered and the filtrate was purified using preparative HPLC to provide compound 49 as a solid. LRMS (+ESI) m/z: 443; 1H NMR: (400 MHz, CD 3 OD) δ 8.46-8.48 (m, 1H), 7.98-8.02(m, 2H), 7.28-7.32 (m, 1H), 6.96-7.01 (m, 2H), 5.35-5.39 (m, 2H), 4.25-4.28 (m, 1H), 3.86-4.06 (m, 4H), 3.61-3.06 (m, 1H), 3.14 (s, 3H), 2.14-2.28 (m, 2H).

Example 15

Preparation of Compound 50

50

Step A- Synthesis of Intermediate Compound 15a

To a solution of 9'-(benzyloxy)-7'-bromo-2'-methyl-2,3,5,6-tetrahydrospiro

[pyran-4,3'-pyrido[l,2-a]pyrazine]- ,8'(2'H,4'H)-dione (30 mg, 0.069 mmol) in 1,4-Dioxane (1 mL) and water (0.1 mL) was added l-(2,4-difluorobenzyl)-4-(4,4,5,5-tetramethyl- 1,3,2- dioxaborolan-2-yl)-lH-pyrazole (66.5 mg, 0.208 mmol), Cs 2 C0 3 (45.1 mg, 0.138 mmol) and Pd(PPh 3 ) 4 (8.00 mg, 6.92 μιηοΐ). The mixture was heated with microwave heating at 130 °C for 1 hours, cooled to room temperature and water (2 mL) was added. The mixture was extracted with ethyl acetate (1 mL x 4) and the combined organic extractss was concentrated in vacuo. The resulting residue was purified using preparative TLC (dichloromethane:methanol, 10: 1) to provide intermediate compound 15a as a white solid. LRMS (+ESI) m/z: 547. 1 HNMR: (300 MHz, CDC1 3 ) δ 8.61 (s, 1H), 7.72 (s, 1H), 7.51-7.53 (m, 3H), 7.15-7.26 (m, 4H), 6.76 (t, / = 8.1 Hz, 2H), 5.26 (s, 2H), 5.24 (s, 2H), 4.09 (s, 2H), 3.81-3.87 (m, 2H), 3.41 (t, /

2.99 (s, 3H), 1.92-2.03 (m, 2H), 1.27-1.32 (m, 2H).

Step B- Synthesis of Compound 50

To a solution of intermediate compound 15a (14 mg, 0.026 mmol) in DMF (2 mL) was added lithium chloride (32.6 mg, 0.768 mmol) at room temperature. The mixture was heated at 90 °C for 3 hours. The mixture was cooled to room temperature and purified using preparative HPLCto provide compound 50 (10 mg, 86 %) as a solid. LRMS (+ESI) m/z: 457.2. 1 HNMR: (400 MHz, DMSO-d 6 ) δ 8.53 (s, 1H), 8.39 (s, 1H), 7.91 (s, 1H), 7.24-7.33 (m, 2H), 7.06-7.09 (m, 1H), 5.40 (s, 2H), 4.54 (s, 2H), 3.81-3.83 (m, 2H), 3.61 (t, / = 11.6 Hz, 2H), 3.05 (s, 3H), 2.08-2.14 (m, 2H), 1.52-1.55 (m, 2H).

Example 16

Preparation of Compound 51

16a 16b

Step A- Synthesis of Intermediate Compound 16a

To a solution of 3-(benzyloxy)-6-((methoxymethoxy)methyl) -4-oxo-4H-pyran-2- carboxylic acid (10.0 g, 32.0 mmol), HATU (24.0 g, 64.0 mmol), HO At (9.0 g, 64.0 mmol) in DMF (80 mL) was added ethanamine (10.0 mL, 153 mmol). The reaction mixture was allowed to stir in a seal tube at room temperature overnight. The reaction mixture was diluted with water (40 mL) and extracted with ethyl acetate (2x 100 mL). The combined organic portion was washed with brine, dried, filtered and evaporated under reduced pressure to provide a residue that was purified using column chromatography on silica gel (petroleum ether: ethyl acetate, 5: 1) to provide intermediate compound 16a as a solid. LRMS (+ESI) m/z = 348.4. 1H NMR (300 MHz, DMSO-J6) δ 8.54-8.58 (m, 1H), 7.34-7.44 (m, 5H), 6.53 (s, 1H), 5.18 (s, 2H), 4.68 (d, / = 3.9 Hz, 2H), 4.46 (d, / = 15.0 Hz, 2H), 3.30 (s, 3H), 3.17-3.27 (m, 2H), 1.02-1.07 (m, 3H).

Step B- Synthesis of Intermediate Compound 16b

To a solution of compound intermediate compound 16a (7.0 g, 20.0 mmol) in EtOH (90 mL) was added NH 3 . water (90 mL) at room temperature, and the mixture was allowed to stir at room temperature for 5 hours. The mixture was concentrated in vacuo to provide crude intermediate compound 16b that was used directly for next step without further purification. LRMS (+ESI) m/z = 347.2. Step C- Synthesis of Intermediate Compound 16c

To a solution of crude intermediate compound 16b (6.0 g, 17 mmol) in DMF (60 mL) was added K 2 C0 3 (4.9 g, 36 mmol) and 0-(2, 4-dinitrophenyl)hydroxylamine (4.0 g, 20 mmol). The reaction mixture was allowed to stir at room temperature for 16 hours. The mixture was directly purified using preparative reverse-phase HPLC to provide intermediate compound 16c as a solid. LRMS (+ESI) m/z = 362.2. 1H NMR (400 MHz, DMSO-J6) δ 8.52-8.54 (m, 1H), 7.28-7.39 (m, 5H), 6.26 (s, 1H), 5.85 (s, 2H), 5.03 (s, 2H), 4.68 (d, J = 4.8 Hz, 2H), 4.58- 4.60 (m, 2H), 3.28 (d, J = 7.2 Hz, 3H), 3.17-3.23 (m, 2H), 1.01-1.05 (m, 3H).

Step D- Synthesis of Intermediate Compound 16d

To a solution of intermediate compound 16c (200 mg, 0.56 mmol) in THF (8 mL) was added Acetic acid (9 mL) and ethyl 4-oxopiperidine-l-carboxylate (2.6 g, 15 mmol) at room temperature and the reaction mixture was heated to 80 °C under microwave heating for 1 hour. The mixture was concentrated in vacuo and the resulting residue was purified using column chromatography on silica gel (EA: CH 3 OH = 120: 1) to provide intermediate compound 16d as a solid. LRMS (+ESI) m/z = 515.2. 1H NMR (400 MHz, CD 3 OD) δ 7.39-7.41 (m, 2H), 7.28-7.35 (m, 3H), 6.69 (s, 1H), 5.26 (d, J = 8.0 Hz, 1H), 5.13 (d, J = 12.0 Hz, 1H), 4.66 (s, 2H), 4.38 (d, J = 12.0 Hz, 1H), 4.28 (d, J = 12.0 Hz, 1H), 4.13-4.19 (m, 2H), 3.90-3.93 (m, 1H), 3.73-3.77 (m, 1H), 3.57 (s, 1H), 3.46 (s, 1H), 3.36 (s, 2H), 3.28-3.32 (m, 1H), 3.22-3.26 (m, 1H), 3.16-3.21 (m, 1H), 2.59-2.66 (m, 2H), 2.24-2.31 (m, 2H), 1.26-1.30 (m, 3H) , 1.06-1.10 (m, 3H).

Step E- Synthesis of Intermediate Compound 16e

To a solution of intermediate compound 16d (200 mg, 0.39 mmol) in DMF (2 mL) was added Cs 2 C0 3 (130 mg, 0.40 mmol) and iodomethane (7 drops). The mixture was allowed to stir for 2 h at room temperature. The reaction mixture was filtered and extracted with ethyl acetate (2x 10 mL). The combined organic extractss were dried over Na 2 S0 4 and concentrated in vacuo to provide intermediate compound 16e, which was used for the next step directly.

LRMS (+ESI) m/z = 529.3. Step F- Synthesis of Intermediate Compound 16f

To a solution of intermediate compound 16e (150 mg, 0.28 mmol) in dichloromethane (2.0 mL) was added N-bromosuccinimide (71 mg, 0.40 mmol) and the mixture was allowed to stir at room temperature for 3 hours. The mixture was concentrated in vacuo and the resulting residue was purified using column chromatography on silica gel (petroleum ether: ethyl acetate = 3: 1 to ethyl acetate) to provide intermediate compound 16f as a solid. LRMS (+ESI) m/z = 607.2. 1H NMR (400 MHz, Methanol-d4) δ 7.36-7.40 (m, 2H), 7.28-7.32 (m, 3H), 5.47 (d, J = 8.0 Hz, 1H), 5.17 (d, J = 12.0 Ηζ,ΙΗ), 5.03 (d, J = 12.0 Hz, 1H), 4.95 (d, J = 8.0 Hz, 1H), 4.71-4.85 (m, 3H), 4.09-4.17 (m, 4H), 3.65-3.73 (m, 2H), 3.54-3.59 (m, 3H), 3.30-3.32 (m, 1H), 2.82 (s, 3H), 2.15-2.31 (m, 2H), 1.38-1.48 (m, 1H), 1.21-1.27 (m, 7H).

Step G- Synthesis of Intermediate Compound 16g

To a solution of intermediate compound 16f (50 mg, 0.08 mmol) in dioxane (3 mL) was added Cs 2 C0 3 (54 mg, 0.16mmol), Pd(PPh 3 ) 4 (20 mg, 0.02 mmol) andl-(2,4- difluorobenzyl -4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrazole (80 mg, 0.25 mmol) at room temperature and the reaction mixture was heated to 130 °C under microwave heating for 1 hour. The mixture was filtered and the filtrate was purified using preparative reverse-phase HPLC to provide intermediate compound 16g as a solid. LRMS (+ESI) m/z = 721.3.

Step H- Synthesis of Intermediate Compound 16h To a solution of intermediate compound 16g (16 mg, 0.02 mmol) in ethyl acetate (4 mL) was added a solution of HCl/ethyl acetate (4 mL) at 0 °C. The mixture was allowed to stir for 3 h at room temperature. The mixture was concentrated in vacuo to provide intermediate compound 16h which was used for the next without purification. LRMS (+ESI) m/z = 677.2.

Step I- Synthesis of Compound 51

To a solution of intermediate compound 16h (12 mg, 0.02 mmol) in dichloromethane (8 mL) was added TFA (3 mL) at 0 °C. The mixture was allowed to stir for 3 hours at room temperature. The mixture was concentrated in vacuo and the resulting residue was purified using preparative HPLC to provide compound 51 as a solid. LRMS (+ESI) m/z = 587.2. 1H NMR (400 MHz, Methanol-d4) 8.06-8.20 (m, 1H), 7.94 (s, 1H), 7.37-7.40 (m, 1H), 6.99-7.06 (m, 2H), 5.37-5.51 (m, 2H), 4.70 (d, J = 12.0 Hz, 1H), 4.26 (d, J = 12.0 Hz, 1H), 4.13- 4.18 (m, 2H), 3.97 (d, J = 12.0 Hz, 1H), 3.84-3.89 (m, 1H), 3.56-3.62 (m, 1H), 3.32-3.33 (m, 3H), 2.91 (s, 3H), 2.30-2.42 (m, 2H), 1.80-1.92 (m, 2H), 1.28-1.33 (m, 6H).

Example 17

Preparation of Compound 52

17c 17 17d Step A - Synthesis of Intermediate Compound 17a

A mixture of intermediate compound 1-2 (100 mg, 0.206 mmol) and tert-butyl 4- amino-4-(aminomethyl)piperidine-l-carboxylate oxalate (65.9 mg, 0.206 mmol) in N- methylimidazole (2 mL) was allowed to stir at room temperature for 2 hours. The reaction mixture was diluted with water (0.5 mL) and directly purified using reverse-phase MPLC to provide intermediate compound 17a as a solid. LCMS (M + H) = 668

Step B - Synthesis of Intermediate Compound 17b A mixture of intermediate compound 17a (60 mg, 0.090 mmol), HATU (51.2 mg, 0.135 mmol) and diisopropylethylamine (34.8 mg, 0.270 mmol) in DMF (1 mL) was allowed to stir at room temperature for 4 hours. The reaction mixture was diluted with water (0.2 mL) and directly purified using reverse-phase HPLC to provide intermediate compound 17b as a solid. LCMS (M + H) = 650

Step C - Synthesis of Intermediate Compound 17c

A mixture of intermediate compound 17b (900 mg, 1.385 mmol), iodomethane (590 mg, 4.16 mmol) and cesium carbonate (2257 mg, 6.93 mmol) in DMF (5 mL) was allowed to stir at room temperature for 1 hour. The reaction mixture was diluted with water (3 mL) and directly purified using reverse-phase MPLC to provide intermediate compound 17c as a solid. LCMS (M + H) = 664

Step D - Synthesis of Intermediate Compound 17 d and Compound 52

A solution of intermediate compound 17c (850 mg, 1.281 mmol) in methanol (10 mL) was treated with a solution of HC1 in methanol (1.25 M, 10.25 ml, 12.81 mmol) at room temperature. The reaction was heated at 60 °C for 2 hours. The solvent was removed in vacuo.

The resulting residue was dissolved in DMSO (5 mL) and purified using reverse-phase MPLC to provide intermediate compound 17d as a solid [LCMS (M + H) = 564] and compound 52 as a solid. 17: 1H NMR (400 MHz, CD 3 OD): δ 8.84 (s, 1H), 7.45 (m, 1H), 6.96-7.02 (m, 2H), 4.73 (s,

2H), 4.48 (s, 2H), 3.51-3.53 (m, 2H), 3.35-3.40 (m, 2H), 3.18 (s, 3H), 2.41-2.44 (m, 2H), 2.05-

2.07 (m, 2H). LCMS (M + H) = 474

Example 18

Preparation of Compound 53

53

Using the methods described in Example 17, and substitute iodethane for iodomethane in Step C, compound 53 was prepared. 1H NMR (400 MHz, CDC1 3 ): δ 8.82 (s, 1H), 7.24-7.40 (m, 1H), 6.81-6.92 (m, 2H), 4.16-4.45 (m, 8H), 3.60-3.68 (m, 2H), 2.97-3.09 (m, 2H), 2.02-2.18 (m, 2H), 1.73-1.83 (m, 2H), 1.26-1.40 (m, 6H). LCMS (M + H) = 560.10.

Example 19

Preparation of Compound 54

Step A - Synthesis of Intermediate Compound 19a

A solution of intermediate compound 17d (380 mg, 0.674 mmol) in THF (7 mL) was cooled at 0 °C and treated with triethylamine (205 mg, 2.023 mmol) and ethyl chloroformate (110 mg, 1.011 mmol). The mixture was allowed to stir at 0 °C for 15 minutes, treated with methanol (3 mL) and stirred at room temperature for 30 minutes. The mixture was concentrated in vacuo and the resulting residue was purified using chromatography on silica gel (5% methanol/dichloromethane) to provide intermediate compound 19a as a solid. LCMS (M + H) = 636.

Step B- Synthesis of Compound 54

Intermediate compound 19a (300 mg, 0.472 mmol) in dichloromethane (2 mL) was treated at room temperature with TFA (4 mL). The mixture was allowed to stir at room temperature for 1 h and then concentrated in vacuo. The resulting residue was purified using reverse-phase HPLC to provide compound 54 as a solid. l U NMR (400 MHz, CDC1 3 ): δ 8.66 (s, 1H), 7.33-7.34 (m, 1H), 6.84-6.87 (m, 2H), 4.45 (s, 2H), 4.44 (s, 2H), 4.27 (m, 2H), 4.19 (q, J = 5.2 Hz, 2H), 3.09 (s, 3H), 3.06 (m, 2H), 2.09 (m, 2H), 1.74 (m, 2H), 1.30 (t, J = 5.2 Hz, 3H). LCMS (M + H) = 546

Step A - Synthesis of Intermediate Compound 20a

A solution of intermediate compound 17b (20 mg, 0.031 mmol) in methanol (1 mL) was treated with a solution of HC1 in methanol ( 1.25 M, 1 ml, 1.250 mmol). The mixture was heated at 60 °C for 2 h and then concentrated in vacuo. The resulting residue was dissolved in methanol (1 mL), diluted with dichloromethane (10 mL) and treated with saturated aqueous NaHCC"3 (2 mL). The mixture was aged at room temperature for 5 minutes, treated with anhydrous Na 2 S0 4 , filtered and the filtrate was concentrated to provide intermediate compound 20a as a solid. LCMS (M + H) = 550.

Step B - Synthesis of Intermediate Compound 20b

A solution of intermediate compound 20a (15 mg, 0.027 mmol) in THF (1 mL) was cooled at 0 °C and treated with triethylamine (7.6 μΐ, 0.06 mmol) and ethyl chloroformate (2.6 μΐ, 0.03 mmol). The reaction was allowed to stir at 0 °C for 15 min and then concentrated in vacuo. The resulting residue was purified using chromatography on silica gel column (8% methanol/dichloromethane) to provide intermediate compound 20b as a solid. LCMS (M + H) = 622. Step C - Synthesis of Compound 55

A solution of intermediate compound 20b (11.5 mg, 0.018 mmol) in dichloromethane (0.5 mL) was treated with TFA (1 mL) and the mixture was allowed to stir at room temperature for 1 h and then concentrated in vacuo. The resulting residue was purified using a reverse-phase HPLC to provide compound 55 as a solid. . 1H NMR (400 MHz, CDCI3): δ 8.70 (s, 1H), 7.55 (s, 1H), 7.31-7.39 (m, 1H), 6.84-6.92 (m, 2H), 4.48 (brs, 2H), 4.24 (brs, 2H), 4.19 (q, J = 7.1 Hz, 2H), 3.77-3.85 (m, 2H), 3.50-3.58 (m, 2H), 1.81-1.92 (m, 4H), 1.30 (t, J = 7.1 Hz, 3H). LCMS (M + H) = 532.14.

Example 21

Preparation of Compound 56

56 Step A - Synthesis of Intermediate Compound 21a

A mixture of compound 3 (100 mg, 0.245 mmol) and 2-((3-(aminomethyl)- tetrahydrofuran-3-yl)amino)ethanol » 2TFA (238 mg, 0.612 mmol) in N-methylimidazole (2.0 ml) heated at 100 °C for 14 hours. The mixture was then cooled to room temperature, diluted with acetonitrile (5 mL) and neutralized with acetic acid. Direct purification of the mixture using preparative reverse-phase HPLC provided intermediate compound 21a. LCMS (M + H) = 505.

1H NMR (500 MHz, CDC1 3 ): δ 8.80 (s; 1 H); 7.30-7.34 (m; 1 H); 6.83-6.88 (m; 2 H); 4.48 (s;

2 H); 4.36 (d; J = 11.9 Hz; 1 H); 4.22 (d; J = 12.3 Hz; 1 H); 4.16 (d; J = 10.1 Hz; 1 H); 4.08 (s;

3 H); 4.03-4.09 (m, 1H); 3.91-4.01 (m; 3 H); 3.77-3.84 (m; 2 H); 3.65 (d; J = 10.5 Hz; 1 H);

2.33 (br s; 1 H); 2.13 (br s; 1 H).

Step B - Synthesis of Compound 56 Intermediate compound 21a (20 mg, 0.04 mmol) was dissolved in DMF (2.0 mL), treated with LiCl (17 mg, 0.40 mmol) and heated at 100 °C for 2 hours. The mixture was cooled to room temperature and directly purified using preparative reverse-phase HPLC to provide compound 56 as a powder. LCMS (M + H) = 491 ; 1H NMR (500 MHz, CDC1 3 ) 8.70 (s; 1 H); 7.36-7.27 (m, 1 H); 6.82-6.92 (m, 2 H); 4.47 (s; 2 H); 4.19-4.31 (m; 3 H); 4.07-4.14 (m; 1 H); 3.88-4.02 (m; 3 H); 3.75-3.79 (m; 2 H); 3.66 (d; J = 10.6 Hz; 1 H). 2.32-2.42 (m; 1 H); 2.14-2.24 (m, 1 H)

Example 22

Preparation of Compound 57 and Intermediate Compound 22a

A mixture of 4-(aminomethyl)tetrahydro-2H-pyran-4-amine » 2HCl (413 mg, 2.04 mmol) and compound 1-2 (1000 mg, 2.06 mmol) in N-methylimidazole (10 mL) was treated with DBU (0.622 mL, 4.13 mmol). The mixture was heated to 100 °C and stirred for 16 hours.

The mixture was cooled to room temperature, diluted with ethyl acetate and washed with water.

The organic portion was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The resulting residue was purified using silica gel chromatography (0-5% methanol/dichloromethane) to provide intermediate compound 22a. LCMS (M + H) = 551.

The aqueous portion was concentrated in vacuo and the resulting residue was purified using reverse-phase HPLC to provide compound 57 LCMS (M + H) = 461.

Example 23

Preparation of Compound 58 and Compound 59

Intermediate compound 22a (500 mg, 0.908 mmol) in DMF (4 mL) was cooled to -20 °C and treated dropwise with LiHMDS (1M in THF, 1.82 mL, 1.82 mmol). The mixture was allowed to stir at -20 °C for 20 minutes, treated with EtI (0.11 mL, 1.36 mmol) and aged for 16 h at -20 °C. The mixture was treated with aqueous IN HC1 (0.2 mL) and the resulting mixture was extracted with ethyl acetate and the combined organic portion was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated in vacuo. The resulting residue was dissolved in TFA (3 mL) and stirred at room temperature for 4 h and then concentrated in vacuo. The resulting residue was directly purified using reverse-phase HPLC to provide compound 58. LCMS (M + H) = 489 and compound 59 LCMS (M + H) = 517.

The following compounds of the present invention were prepared using the method described in the Example above using the appropriate reactants and reagents.

Example 24

Assessing antiviral potency in a multiple round HIV-1 infection assay

HIV-1 replication was monitored using MT4-gag-GFP clone D3 (hereafter designate MT4-GFP), which are MT-4 cells modified to harbor a GFP reporter gene, the expression of which is dependent on the HIV-1 expressed proteins tat and rev. Productive infection of an MT4-GFP cell with HIV-1 results in GFP expression approximately 24h postinfection.

MT4-GFP cells were maintained at 37°C/5% CO 2 /90% relative humidity in RPMI 1640 supplemented with 10% fetal bovine serum, 100 U/ml penicillin/streptomycin, and

400μg/ml G418 to maintain the reporter gene. For infections, MT4-GFP cells were placed in the same medium lacking G418 and infected overnight with H9IIIB or NL4-3 virus at an approximate multiplicity of infection of 0.01 in the same incubation conditions. Cells were then washed and resuspended in either RPMI 1640 containing no serum at 1.6 x 10 5 cells/mL (serum free conditions), 10% normal human serum at 1.6 x 10 5 cells/mL (10% NHS conditions) or in 100% normal human serum at 2 x 10 5 cells/mL (100% NHS conditions). Compound plates were prepared by dispensing compounds dissolved in DMSO into wells of 384 well poly D lysine-coated plates (0.2μ1Λνε11) using an ECHO acoustic dispenser. Each compound was tested in a 10 point serial 3-fold dilution (typical final concentrations: 4.2 μΜ - 0.21 nM). Controls included no inhibitor (DMSO only) and a combination of three antiviral agents (efavirenz, indinavir, and the integrase strand transfer inhibitor L-002254051 at final concentrations of 4μΜ each). Cells were added (50μΕΛνε11) to compound plates and the infected cells were maintained at 37°C/5% CO 2 /90% relative humidity.

Infected cells were quantified at two time points, ~48h and- 72h post-infection, by counting the number of green cells in each well using an Acumen eX3 scanner. The increase in the number of green cells over ~24h period gives the reproductive ratio, Ro, which is typically 5-15 and has been shown experimentally to be in logarithmic phase (data not shown). Inhibition of Ro is calculated for each well, and IC50S determined by non-linear 4-parameter curve fitting.

ViKinG IP

compound ViKinG IP (nM) ViKinG IP (nM)

(nM) with 0%

# with 10% NHS with 100% NHS

NHS

5 16 22 250

6 NA 13 159

7 30 58 148

8 6 NA 131

9 90 NA 344

10 NA 57 1499

11 NA 31 359

12 NA 24 126

13 9 14 106

14 NA 41 NA

15 8 178 3135

16 17 47 NA

17 29 NA 1518

18 NA 13 254

19 NA 9 133 20 NA 234 NA

21 NA 141 NA

22 NA 47 731

23 NA 100 NA

24 12 6 86

25 5 17 257

26 14 26 139

27 21 72 712

28 54 122 679

29 89 NA 2075

30 38 NA 280

31 NA 121 2099

32 8 NA 74

33 NA 49 89

34 10 34 NA

35 2 NA 133

36 4 NA 368

37 5 NA 456

38 4 NA 132

39 2 NA 42

40 4 NA 49

41 7 NA 3602

42 2 NA 36

43 2 NA 26

44 11 NA 7804

45 3 NA 236

46 125 NA 194

47 2 NA 101

48 4 NA 118

49 34 NA 123

50 60 NA 181

51 159 NA 3646

52 74 114 NA

53 12 NA 104

54 11 NA 116

55 69 NA 2241 56 42 NA 1051

57 92 NA 2032

58 41 NA 870

59 5 NA 70

60 29 NA 704

61 7 NA 85

NA = not available