Antiviral indoleoxoacetyl piperazine derivatives

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention provides compounds having drug and bio-affecting properties, their pharmaceutical compositions and method of use. In particular, the invention is concerned with indoleoxoacetyl piperazine derivatives. These compounds possess unique antiviral activity, whether used alone or in combination with other antivirals, antiinfectives, immunomodulators or HIV entry inhibitors. More particularly, the present invention relates to the treatment of HIV and AIDS.

2. Background Art

HIV-1 (human immunodeficiency virus-1) infection remains a major medical problem, with an estimated 33.4 million people infected worldwide. Currently available HIV drugs include six nucleoside reverse transcriptase (RT) inhibitors (zidovudine, didanosine, stavudine, lamivudine, zalcitabine and abacavir), three non-nucleoside reverse transcriptase inhibitors (nevirapine, delavirdine and efavirenz) as well as five peptidomimetic protease inhibitors (saquinavir, indinavir, ritonavir, nelfinavir and amprenavir). Each of these drugs can only transiently restrain viral replication if used alone. However, when used in combination, these drugs have a profound effect on disease progression. In fact, significant reductions in death rates among AIDS patients have been recently documented. Despite these results, 30 to 50% of patients ultimately fail combination drug therapies. Insufficient drug potency, non-compliance, restricted tissue penetration and drug-specific limitations within certain cell types (e.g. most nucleoside analogs cannot be phosphorylated in resting cells) may account for the incomplete suppression of sensitive viruses. Furthermore, the high replication rate and rapid turnover of HIV-1 combined with the frequent incorporation of mutations, leads to the appearance of drug-resistant variants and treatment failures when suboptimal drug concentrations are present (Larder and Kemp, Gulick, Morris-Jones, et al, Kuritzkes, Vacca and Condra, Schinazi, et al and Flexner, Ref. 6-12). Therefore, novel anti-HIV agents exhibiting distinct resistance patterns, and favorable pharmacokinetic as well as safety profiles are needed to provide more treatment options.

Currently marketed HIV-1 drugs are dominated by either nucleoside reverse transcriptase inhibitors or peptidomimetic protease inhibitors. Non-nucleoside reverse transcriptase inhibitors have recently gained an increasingly important role in the therapy of HIV infections. At least 30 different classes of NNRTIs have been published in the literature (DeClercq, Ref. 13). Dipyridodiazepinone (nevirapine), benzoxazinone (efavirenz) and bis(heteroaryl) piperazine derivatives (delavirdine) are already approved for clinical use. In addition, several indole derivatives including indole-3-sulfones, piperazino indoles, pyrazino indoles, and 5H-indolo[3,2-b][1,5]benzothiazepine derivatives have been reported as HIV-1 reverse transciptase inhibitors (Greenlee et al, Ref. 1, Williams et al, Ref. 2, Romero et al, Ref. 3, Font et al, Ref. 14, Romero et al, Ref. 15, Young et al, Ref. 16, Genin et al, Ref. 17, and Silvestri et al, Ref. 18). Indole 2-carboxamides have also been described as inhibitors of cell adhesion and HIV infection (Boschelli et al. in U.S. Pat. No. 5,424,329, Ref. 4). Finally, 3-substituted indole natural products (Semicochliodinol A and B, didemethylasterriquinone and isocochliodinol) were disclosed as inhibitors of HIV-1 protease (Fredenhagen et al, Ref. 19). However, nothing in these references can be construed to disclose or suggest the novel compounds of this invention and their use to inhibit antiviral infection, including HIV infection.

Structurally related compounds have been disclosed previously (Brewster et al, Ref. 20, Archibald et al, Ref. 21, American Home Products in GB 1126245, Ref. 5). However, the structures differ from those claimed herein in that they are symmetrical bis(3-indolylglyoxamides) rather than unsymmetrical aroyl indoleoxoacetyl piperazine derivatives, and there is no mention of use for treating antiviral infections. Interestingly, the indole moiety present in the compounds disclosed here is the common feature of many non-nucleoside HIV-1 reverse transcriptase inhibitors including Delavirdine from Upjohn (Dueweke et al. 1992, 1993, Ref. 22 and 23).

Additionally, the following compounds are available commercially but have not been reported as being useful as pharmaceuticals, and more specifically for antiviral use in mammals.

Compound LJ952 (available from Menai Organics Ltd., Gwynedd, North Wales):

Compound TRI-29586 (available from Tripos):

REFERENCES CITED

Patent Documents

1. Greenlee, W. J.; Srinivasan, P. C., Indole reverse transcriptase inhibitors. U.S. Pat. No. 5,4124,327.

2. Williams, T. M.; Ciccarone, T. M.; Saari, W. S.; Wai, J. S.; Greenlee, W. J.; Balani, S. K.; Goldman, M. E.; Theohrides, A. D., Indoles as inhibitors of HIV reverse transcriptase. European Patent 530907.

3. Romero, D. L.; Thomas, R. C., Preparation of substituted indoles as anti-AIDS pharmaceuticals. PCT WO 93/01181.

4. Boschelli, D. H.; Connor, D. T.; Unangst, P. C., Indole-2-carboxamides as inhibitors of cell adhesion. U.S. Pat. No. 5,424,329.

5. Therapeutic bis(indolyl) compounds. British Patent 1126245 (American Home Products Corp.).

OTHER PUBLICATIONS

6. Larder B. A & Kemp S. D., Multiple mutations in the HIV-1 reverse transcriptase confer high-level resistance to zidovudine (AZT),

Science

, 246:1155-1158, 1989.

7. Gulick R. M., Current antiretroviral therapy: an overview.,

Quality of Life Research

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8. Kuritzkes D. R., HIV resistance to current therapies,

Antiviral Therapy

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9. Morris-Jones S, Moyle G & Easterbrook P. J., Antiretroviral therapies in HIV-1 infection,

Expert Opinion on Inzestigational Drugs

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10. Schinazi R. F, Larder B. A & Mellors J. W., Mutations in retroviral genes associated with drug resistance,

International Antiviral News

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11. Vacca J. P & Condra J. H., Clinically effective HIV-1 protease inhibitors,

Drug Discovery Today

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12. Flexner D., HIV-protease inhibitors,

Drug Therapy

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13. De Clercq E., The role of non-nucleoside reverse transcriptase inhibitors (NNRTIs) in the therapy of HIV-1 infection, Antiviral Research Vol. 38 pp. 153-179, 1998.

14. Font, M.; Monge, A.; Cuartero, A.; Elorriaga, A.; Martinez-Irujo, J. J.; Alberdi, E.; Santiago, E.; Prieto, I.; Lasarte, J. J.; Sarobe, P. and Borras, F., Indoles and pyrazino[4,5-b]indoles as nonnucleoside analog inhibitors of HIV-1 reverse transcriptase, Eur. J. Med. Chem., 30, 963-971, 1995.

15. Romero, D. L.; Morge, R. A.; Genin, M. J.; Biles, C.; Busso, M,; Resnick, L.; Althaus, I. W.; Reusser, F.; Thomas, R. C and Tarpley, W. G., Bis(heteroaryl)piperazine (BHAP) reverse transcriptase inhibitors: structure-activity relationships of novel substituted indole analogues and the identification of 1-[(5-methanesulfonamido-1H-indol-2-yl)-carbonyl]-4-[3-[1-methylethyl)amino]-pyridinyl]piperazine momomethansulfonate (U-90152S), a second generation clinical candidate, J. Med. Chem., 36, 1505-1508,1993.

16. Young, S. D.; Amblard, M. C.; Britcher, S. F.; Grey, V. E.; Tran, L. O.; Lumma, W. C.; Huff, J. R.; Schleif, W. A.; Emini, E. E.; O'Brien, J. A.; Pettibone, D. J. 2-Heterocyclic indole-3-sulfones as inhibitors of HIV-reverse transcriptase, Bioorg. Med. Chem. Lett, 5, 491-496, 1995.

17. Genin, M. J.; Poel, T. J.; Yagi, Y.; Biles, C.; Althaus, I.; Keiser, B. J.; Kopta, L. A.; Friis, J. M.; Reusser, F.; adams, W. J.; Olmsted, R. A.; Voorman, R. L.; Thomas, R. C. and Romero, D. L., Synthesis and bioactivity of novel bis(heteroaryl)piperazine (BHAP) reverse transcriptase inhibitors: structure-activity relationships and increased metabolic stability of novel substituted pyridine analogs, J. Med. Chem., 39, 5267-5275,1996.

18. Silvestri, R.; Artico, M.; Bruno, B.; Massa, S.; Novellino, E.; Greco, G.; Marongiu, M. E.; Pani, A.; De Montis, A and La Colla, P., Synthesis and biological evaluation of 5H-indolo[3,2-b][1,5]benzothiazepine derivatives, designed as conformationally constrained analogues of the human immunodeficiency virus type 1 reverse transciptase inhibitor L-737,126. Antiviral Chem. Chemother., 9, 139-148, 1998.

19. Fredenhagen, A.; Petersen, F.; Tintelnot-Blomley, M.; Rosel, J.; Mett, H and Hug, P. J., Semicochliodinol A and B: inhibitors of HIV-1 protease and EGF-R protein Tyrosine Kinase related to Asterriquinones produced by the fungus

Chrysosporium nerdarium

, Antibiotics, 50, 395-401, 1997.

20. Brewster, K.; Green, D. M.; Pinder, R. M.; Thompson, P. B. J., Antihypertensive 1,4-bis(2-indol-3-ylethyl)piperazines,

Chim. Ther

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21. Archibald, John L.; Freed, Meier E., 1,4-Bis(2-indol-3-ylethyl)piperazines,

J. Med Chem

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22. Dueweke T. J. et al, The binding of a novel bisheteroaryliperazine mediates inhibition of human immunodeficiency virus type 1 reverse transcriptase, J Biol. Chem. Vol. 267 pp27-30,1992.

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24. Gribble, G. W., Recent developments in indole ring synthesis-methodology and applications,

Contemp. Org. Synth

., 1, 145-72 1994.

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Justus Liebigs Ann. Chem

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26. Desai, M.; Watthey, J. W. H.; Zuckerman, M., A convenient preparation of 1-aroylpiperazines, Org. Prep. Proced. Int., 8, 85-6, 1976.

27. Potts, B. J., Mini Reverse transcriptase (RT) assay, In Aldovini A., B. D. Walker (ed), Techniques in HIV Research, Stockton Press, NY, p. 103-106, 1990.

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29. Johnson, V. A. and R. E. Byrington, Infectivity assay, p. 71-76 in A. Aldovini and B. D. Walker (ed), techniques in HIV Research, Stockton Press, New York, 1990.

30. Harada, S., Koyanagi, Y., and N. Yamamoto, Infection of HTLV-III/LAV in HTLV-I carrying cells MT-2 and MT-4 and application in a plaque assay, Science 229:563-566, 1985.

31. (a) Behun, J. D.; Levine, R.

J. Org. Chem

. 1961, 26, 3379. (b) Rossen, K.; Weissman, S. A.; Sager, J.; Reamer, R. A.; Askin, D.; Volante, R. P.; Reider, P. J. Asymmetric Hydrogenation of tetrahydropyrazines: Synthesis of (S)-piperazine 2-tert-butylcarboxamide, an intermediate in the preparation of the HIV protease inhibitor Indinavir.

Tetrahedron Lett

., 1995, 36, 6419-6422. (c) Jenneskens, L. W.; Mahy, J.; den Berg, E. M. M. de B.-v.; Van der Hoef, I.; Lugtenburg, J.

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Bas

1995, 114, 97.

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J. Org. Chem

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Org. Prep. Proced. Int

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SUMMARY DESCRIPTION OF THE INVENTION

It has now been surprisingly found that compounds of formula I, or pharmaceutically acceptable salts thereof, are effective antiviral agents, particularly for treating HIV, whether used alone or in combination with other antivirals, antiinfectives, immunomodulators or HIV entry inhibitors.

The present invention comprises compounds of formula I, or pharmaceutically acceptable salts thereof,

wherein:

R

1

, R

2

, R

3

, R

4

and R

5

are each independently H, C

1

-C

6

alkyl, C

3

-C

6

cycloalkyl, C

2

-C

6

alkenyl, C

3

-C

6

cycloalkenyl, C

2

-C

6

alkynyl, halogen, CN, nitro, COOR

6

or XR

7

, each of said alkyl and cycloalkyl being optionally substituted with one to three same or different halogen, amino, OH, CN or NO

2

;

R

6

is H, C

1

-C

6

alkyl, or C

3

-C

6

cycloalkyl, benzyl, each of said alkyl, cycloalkyl and benzyl being optionally substituted with one to three same or different halogen, amino, OH, CN or NO

2

;

X is O, S or NR

6

R

7

;

R

7

is H, C

1

-C

6

alkyl, C

3

-C

6

cycloalkyl, C

2

-C

6

alkenyl, C

3

-C

6

cycloalkenyl, C

2

-C

6

alkynyl or C(O)R

8

, each of said alkyl and cycloalkyl being optionally substituted with one to three same or different halogen, OH, amino, CN or NO

2

;

R

8

is H, C

1

-C

6

alkyl or C

3

-C

6

cycloalkyl;

—W— is

R

9

, R

10

, R

11

, R

12

, R

13

, R

14

, R

15

, R

16

, R

17

, R

18

, R

19

, R

20

, R

21

, R

22

are each independently H, C

1

-C

6

alkyl, C

3

-C

6

cycloalkyl, C

2

-C

6

alkenyl, C

3

-C

6

cycloalkenyl, C

2

-C

6

alkynyl, CR

23

R

24

OR

25

, COR

26

, COOR

27

or C(O)NR

28

R

29

, each of said alkyl and cycloalkyl being optionally substituted with one to three same or different halogen, amino, OH, CN or NO

2

;

R

23

, R

24

, R

25

, R

26

, R

27

, R

28

, R

29

are each independently H, C

1

-C

6

alkyl, C

3

-C

6

cycloalkyl, C

2

-C

6

alkenyl, C

3

-C

6

cycloalkenyl or C

2

-C

6

alkynyl;

Ar is a 4-7 membered aromatic ring which may contain one to five heteroatoms independently selected from the group consisting of O, S, N or NR

6

, wherein said aromatic ring is optionally fused to group B;

B is an aromatic group selected from the group consisiting of phenyl, 1-naphthyl, 2-naphthyl, indenyl, azulenyl, fluorenyl, and anthracenyl; or a heteroaryl group selected from the group consisting of 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, 1,2,3-oxadiazolyl, 1,2,3-triazolyl, 1,3,4-thiadiazolyl, pyridazinyl, pyrimnidinyl, pyrazinyl, 1,3,5-triazinyl, 1,3,5-trithianyl, indolizinyl, indolyl, isoindolyl, 3H-indolyl, indolinyl, benzo[b]furanyl, benzo[b]thiophenyl, 1H-indazolyl, benzimidazolyl, benzthiazolyl, purinyl, 4H-quinolizinyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 1,8-naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, and phenoxazinyl;

B and said 4-7 membered aromatic ring may each independently contain one to five substituents which are each independently selected from R

30

R

31

, R

32

, R

33

or R

34

;

R

a

and R

b

are each independently H, C

1-6

alkyl or phenyl;

Z is 4-methoxyphenyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, pyrazyl, quinolyl, 3,5-dimethylisoxazoyl, isoxazoyl, 2-methylthiazoyl, thiazoyl, 2-thienyl, 3-thienyl, or pyrimidyl; and p is 0-2;

R

30

R

31

, R

32

, R

33

, and R

34

are each independently H, C

1

-C

6

alkyl, C

3

-C

6

cycloalkyl, C

2

-C

6

alkenyl, C

3

-C

6

cycloalkenyl, C

2

-C

6

alkynyl, halogen, CN, nitro, C(O)R

35

, COXR

36

, hydroxyl, COOR

6

, hydroxymethyl, trifluoromethyl, trifluoromethoxy, O—[(C

1

-C

4

)-straight or branched alkyl], O-benzyl, O-phenyl, 1,2-methylenedioxy, OC(O)C

1-6

alkyl, SC(O)C

1-6

alkyl, S(O)

m

C

1-6

alkyl, S(O)

2

NR

a

R

b

, amino, carboxyl, O-Z, CH

2

—(CH

2

)

p

-Z, O—(CH

2

)

p

-Z, (CH

2

)

p

—O-Z, CH═CH-Z or XR

37

, each of said alkyl and cycloalkyl being optionally substituted with one to three same or different halogen, amino, OH, CN or NO

2

;

m is 0-2;

R

35

and R

36

are each independently H, C

1

-C

6

alkyl or C

3

-C

6

cycloalkyl;

R

37

is H, C

1

-C

6

alkyl, C

3

-C

6

cycloalkyl, C

2

-C

6

alkenyl, C

3

-C

6

cycloalkenyl, C

2

-C

6

alkynyl, C(O)R

38

or C(O)OR

39

, each of said alkyl and cycloalkyl being optionally substituted with one to three same or different halogen, amino, OH, CN or NO

2

;

R

38

, R

39

are each independently H, C

1

-C

6

alkyl or C

3

-C

6

cycloalkyl, each of said alkyl and cycloalkyl being optionally substituted with one to three same or different halogen, amino, OH, CN or NO

2

; provided R

39

is not H;

R

40

is (CH

2

)

n

—Y, where n is 0-6;

Y is selected from:

(1) H, C

1-6

alkyl, C

3-6

cycloalkyl, C

2-6

alkenyl, C

3-6

cycloalkenyl, C

2-6

alkynyl, halogen, CN, nitro, Ar, COOR

6

, COOAr, —CONR

a

R

b

, TR

6

, NR

a

R

b

, —NC(O)NR

a

R

b

, —OC(O)R

6

, —C[N(R

a

)

2

]=N-T-R

b

, XR

6

, —C(O)R

6

, —C(O)Ar, —S(O)R

a

or —S(O)

2

R

a

, provided when Y is —S(O)R

a

or —S(O)

2

R

a

then R

a

is not H; and

(2) a 4-7 membered heterocyclic ring, optionally substituted with R

6

, which may contain 1-3 heteroatoms selected from the group consisting of O, S, SO, SO

2

, N, and NR

41

, wherein R

41

is selected from the group consisting of hydrogen, (C

1

-C

4

)-straight or branched alkyl, (C

2

-C

4

)-straight or branched alkenyl or alkynyl;

T is S or O;

provided R

1

-R

5

, R

9

-R

16

and R

30

-R

34

are not all H at the same time and Ar is phenyl; and

provided R

1

-R

5

, R

9

-R

16

and R

30

-R

34

are not all H at the same time and Ar is 2-furyl.

Another embodiment of the invention is a pharmaceutical formulation which comprises an antiviral effective amount of a compound of formula I.

Another embodiment of the invention is a pharmaceutical formulation useful for treating infection of HIV which additionally comprises an antiviral effective amount of an AIDS treatment agent selected from the group consisting of:

(a) an AIDS antiviral agent;

(b) an anti-infective agent;

(c) an immunomodulator; and

(d) HIV entry inhibitors.

Another embodiment of the invention is a method for treating mammals infected with a virus (e.g. HIV), comprising administering to said mammal an antiviral effective amount of a compound of formula II, or pharmaceutically acceptable salts thereof,

R

9

, R

10

, R

11

, R

12

, R

13

, R

14

, R

15

, R

16

, R

17

, R

18

, R

19

, R

20

, R

21

, R

22

are each independently H, C

1

-C

6

alkyl, C

3

-C

6

cycloalkyl, C

2

-C

6

alkenyl, C

3

-C

6

cycloalkenyl, C

2

-C

6

alkynyl, CR

23

R

24

OR

25

, COR

26

, COOR

27

or C(O)NR

28

R

29

, each of said alkyl and cycloalkyl being optionally substituted with one to three same or different halogen, amino, OH, CN or NO

2

;

R

23

, R

24

, R

25

, R

26

, R

27

, R

28

, R

29

are each independently H, C

1

-C

6

alkyl, C

3

-C

6

cycloalkyl, C

2

-C

6

alkenyl, C

3

-C

6

cycloalkenyl or C

2

-C

6

alkynyl;

Ar is a 4-7 membered aromatic ring which may contain one to five heteroatoms independently selected from the group consisting of O, S, N or NR

6

, wherein said aromatic ring is optionally fused to group B;

B is an aromatic group selected from the group consisting of phenyl, 1-naphthyl, 2-naphthyl, indenyl, azulenyl, fluorenyl, and anthracenyl; or a heteroaryl group selected from the group consisting of 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, 1,2,3-oxadiazolyl, 1,2,3-triazolyl, 1,3,4-thiadiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,3,5-triazinyl, 1,3,5-trithianyl, indolizinyl, indolyl, isoindolyl, 3H-indolyl, indolinyl, benzo[b]furanyl, benzo[b]thiophenyl, 1H-indazolyl, benzimidazolyl, benzthiazolyl, purinyl, 4H-quinolizinyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 1,8-naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, and phenoxazinyl;

B and said 4-7 membered aromatic ring may each independently contain one to five substituents which are each independently selected from R

3

o R

31

, R

32

, R

33

or R

34

;

R

a

and R

b

are each independently H, C

1-6

alkyl or phenyl;

Z is 4-methoxyphenyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, pyrazyl, quinolyl, 3,5-dimethylisoxazoyl, isoxazoyl, 2-methylthiazoyl, thiazoyl, 2-thienyl, 3-thienyl, or pyrimidyl; and p is 0-2;

R

30

R

31

, R

32

, R

33

, and R

34

are each independently H, C

1

-C

6

alkyl, C

3

-C

6

cycloalkyl, C

2

-C

6

alkenyl, C

3

-C

6

cycloalkenyl, C

2

-C

6

alkynyl, halogen, CN, nitro, C(O)R

35

, COXR

36

, hydroxyl, COOR

6

, hydroxymethyl, trifluoromethyl, trifluoromethoxy, O—[(C

1

-C

4

)-straight or branched alkyl], O-benzyl, O-phenyl, 1,2-methylenedioxy, OC(O)C

1-6

alkyl, SC(O)C

1-6

alkyl, S(O)

m

C

1-6

alkyl, S(O)

2

NR

a

R

b

, amino, carboxyl, O-Z, CH

2

—(CH

2

)

p

-Z, O-(CH

2

)

p

-Z, (CH

2

)

p

-O-Z, CH═CH-Z or XR

37

, each of said alkyl and cycloalkyl being optionally substituted with one to three same or different halogen, amino, OH, CN or NO

2

;

m is 0-2;

R

35

and R

36

are each independently H, C

1

-C

6

alkyl or C

3

-C

6

cycloalkyl;

R

37

is H, C

1

-C

6

alkyl, C

3

-C

6

cycloalkyl, C

2

-C

6

alkenyl, C

3

-C

6

cydloalkenyl, C

2

-C

6

alkynyl, C(O)R

38

or C(O)OR

39

, each of said alkyl and cycloalkyl being optionally substituted with one to three same or different halogen, amino, OH, CN or NO

2

;

R

38

, R

39

are each independently H, C

1

-C

6

alkyl or C

3

-C

6

cycloalkyl, each of said alkyl and cycloalkyl being optionally substituted with one to three same or different halogen, amino, OH, CN or NO

2

; provided R

39

is not H;

R

40

is (CH

2

)

n

—Y, where n is 0-6;

Y is selected from:

(1) H, C

1-6

alkyl, C

3-6

cycloalkyl, C

2-6

alkenyl, C

3-6

cycloalkenyl, C

2-6

alkynyl, halogen, CN, nitro, Ar, COOR

6

, COOAr, —CONR

a

R

b

, TR

6

, NR

a

R

b

, —NC(O)NR

a

R

b

, —OC(O)R

6

, —C[N(R

a

)

2

]=N-T-R

b

, XR

6

, —C(O)R

6

, —C(O)Ar, —S(O)R

a

or S(O)

2

R

a

, provided when Y is —S(O)R

a

or —S(O)

2

R

a

then R

a

is not H; and

(2) a 4-7 membered heterocyclic ring, optionally substituted with R

6

, which may contain 1-3 heteroatoms selected from the group consisting of O, S, SO, SO

2

, N, and NR

41

, wherein R

41

is selected from the group consisting of hydrogen, (C

1

-C

4

)-straight or branched alkyl, (C

2

-C

4

)-straight or branched alkenyl or alkynyl; and

T is S or O.

In a preferred embodiment, compounds of formula I and II include those where Ar is phenyl, furyl, isoxazolyl, thiophenyl, pyrazolyl, pyridyl, benzofuryl, benzothiophenyl, indolyl, pyrazinyl, thiazolyl, imidazolyl, thiadiazolyl.

Also preferred are compounds of formulas I and II wherein

W is

R

9

, R

10

, R

11

, R

12

, R

13

, R

14

, and R

15

are each H; and

R

16

is methyl.

Also preferred are compounds of formulas I and II wherein R

2

is H, fluoro or methoxy.

Also preferred are compounds of formulas I and II wherein R

1

, R

3

and R

4

are each H.

DETAILED DESCRIPTION OF THE INVENTION

The synthesis procedures and anti-HIV-1 activities of the novel indoleoxoacetyl piperazine analogs of formula I are summarized below.

Chemistry

The present invention comprises compounds of formula I, their pharmaceutical formulations, and their use in patients suffering from or susceptible to HIV. The compounds of formula I which include pharmaceutically acceptable salts thereof,

The term “C

1-6

alkyl” as used herein and in the claims (unless the context indicates otherwise) mean straight or branched chain alkyl groups such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, amyl, hexyl and the like. Similarly, “C

2-6

alkenyl” and “C

2-6

alkynyl” include straight or branched chain groups.

The term “pharmaceutically acceptable salt” as used herein and in the claims is intended to include nontoxic base addition salts. Suitable salts include those derived from organic and inorganic acids such as, without limitation, hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, methanesulfonic acid, acetic acid, tartaric acid, lactic acid, sulfinic acid, citric acid, maleic acid, fumaric acid, sorbic acid, aconitic acid, salicylic acid, phthalic acid, and the like.

“Halogen” refers to chlorine, bromine, iodine or fluorine.

In the method of the present invention, the term “antiviral effective amount” means the total amount of each active component of the method that is sufficient to show a meaningful patient benefit, i.e., healing of acute conditions characterized by inhibition of antiviral infection, including HIV infection. When applied to an individual active ingredient, administered alone, the term refers to that ingredient alone. When applied to a combination, the term refers to combined amounts of the active ingredients that result in the desired antiviral effect, whether administered in combination, serially or simultaneously. The terms “treat, treating, treatment” as used herein and in the claims means preventing or ameliorating diseases associated with viral infection, including HIV infection.

The present invention is also directed to combinations of the compounds with one or more agents useful in the treatment of AIDS. 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 the AIDS antivirals,

ANTIVIRALS

Drug Name

Manufacturer

Indication

097

Hoechst/Bayer

HIV infection,

AIDS, ARC

(non-nucleoside

reverse tran-

scriptase (RT)

inhibitor)

Amprenivir

Glaxo Wellcome

HIV infection,

141 W94

AIDS, ARC

GW 141

(protease inhibitor)

Abacavir (1592U89)

Glaxo Wellcome

HIV infection,

GW 1592

AIDS, ARC

(RT inhibitor)

Acemannan

Carrington Labs

ARC

(Irving, TX)

Acyclovir

Burroughs Wellcome

HIV infection, AIDS,

ARC, in combination

with AZT

AD-439

Tanox Biosystems

HIV infection, AIDS,

ARC

AD-519

Tanox Biosystems

HIV infection, AIDS,

ARC

Adefovir dipivoxil

Gilead Sciences

HIV infection

AL-721

Ethigen

ARC, PGL

(Los Angeles, CA)

HIV positive, AIDS

Alpha Interferon

Glaxo Wellcome

Kaposi's sarcoma,

HIV in combination

w/Retrovir

Ansamycin

Adria Laboratories

ARC

LM 427

(Dublin, OH)

Erbamont

(Stamford, CT)

Antibody which

Advanced Biotherapy

AIDS, ARC

Neutralizes pH

Concepts

Labile alpha aberrant

(Rockville, MD)

Interferon

AR177

Aronex Pharm

HIV infection, AIDS,

ARC

Beta-fluoro-ddA

Nat'l Cancer Institute

AIDS-associated

diseases

BMS-232623

Bristol-Myers Squibb/

HIV infection,

(CGP-73547)

Novartis

AIDS, ARC

(protease inhibitor)

BMS-234475

Bristol-Myers Squibb/

HIV infection,

(CGP-61755)

Novartis

AIDS, ARC

(protease inhibitor)

CI-1012

Warner-Lambert

HIV-1 infection

Cidofovir

Gilead Science

CMV retinitis,

herpes,

papillomavirus

Curdlan sulfate

AJI Pharma USA

HIV infection

Cytomegalovirus

MedImmune

CMV retinitis

Immune globin

Cytovene

Syntex

Sight threatening

Ganciclovir

CMV

peripheral CMV

retinitis

Delaviridine

Pharmacia-Upjohn

HIV infection,

AIDS, ARC

(RT inhibitor)

Dextran Sulfate

Ueno Fine Chem.

AIDS, ARC, HIV

Ind. Ltd. (Osaka,

positive

Japan)

asymptomatic

ddC

Hoffman-La Roche

HIV infection, AIDS,

Dideoxycyfidine

ARC

ddI

Bristol-Myers Squibb

HIV infection, AIDS,

Dideoxyinosine

ARC; combination

with AZT/d4T

DMP-450

AVID

HIV infection,

(Camden, NJ)

AIDS, ARC

(protease inhibitor)

Efavirenz

DuPont Merck

HIV infection,

(DMP 266)

AIDS, ARC

(-)6-Chloro-4-(S)-

(non-nucleoside RT

cyclopropylethynyl-

inhibitor)

4(S)-trifluoro-

methyl-1,4-dihydro-

2H-3,1-benzoxazin-

2-one, STOCRINE

EL10

Elan Corp, PLC

HIV infection

(Gainesville, GA)

Famciclovir

Smith Kline

herpes zoster,

herpes simplex

FTC

Emory University

HIV infection,

AIDS, ARC

(reverse transcriptase

inhibitor)

GS 840

Gilead

HIV infection,

AIDS, ARC

(reverse transcriptase

inhibitor)

HBY097

Hoechst Marion

HIV infection,

Roussel

AIDS, ARC

(non-nucleoside

reverse transcriptase

inhibitor)

Hypericin

VIMRx Pharm.

HIV infection, AIDS,

ARC

Recombinant Human

Triton Biosciences

AIDS, Kaposi's

Interferon Beta

(Almeda, CA)

sarcoma, ARC

Interferon alfa-n3

Interferon Sciences

ARC, AIDS

Indinavir

Merck

HIV infection, AIDS,

ARC, asymptomatic

HIV positive, also in

combination with

AZT/ddI/ddC

ISIS 2922

ISIS Pharmaceuticals

CMV retinitis

KNI-272

Nat'l Cancer Institute

HIV-assoc. diseases

Lamivudine, 3TC

Glaxo Wellcome

HIV infection,

AIDS, ARC

(reverse

transcriptase

inhibitor); also

with AZT

Lobucavir

Bristol-Myers Squibb

CMV infection

Nelfinavir

Agouron

HIV infection,

Pharmaceuticals

AIDS, ARC

(protease inhibitor)

Nevirapine

Boeheringer

HIV infection,

Ingleheim

AIDS, ARC

(RT inhibitor)

Novapren

Novaferon Labs, Inc.

HIV inhibitor

(Akron, OH)

Peptide T

Peninsula Labs

AIDS

Octapeptide

(Belmont, CA)

Sequence

Trisodium

Astra Pharm.

CMV retinitis, HIV

Phosphonoformate

Products, Inc.

infection, other CMV

infections

PNU-440690

Pharmacia Upjohn

HIV infection,

AIDS, ARC

(protease inhibitor)

Probucol

Vyrex

HIV infection, AIDS

RBC-CD4

Sheffield Med.

HIV infection,

Tech (Houston, TX)

AIDS, ARC

Ritonavir

Abbott

HIV infection,

AIDS, ARC

(protease inhibitor)

Saquinavir

Hoffmann-

HIV infection,

LaRoche

AIDS, ARC

(protease inhibitor)

Stavudine; d4T

Bristol-Myers Squibb

HIV infection, AIDS,

Didehydrodeoxy-

ARC

thymidine

Valaciclovir

Glaxo Wellcome

Genital HSV & CMV

infections

Virazole

Viratek/ICN

asymptomatic HIV

Ribavirin

(Costa Mesa, CA)

positive, LAS, ARC

VX-478

Vertex

HIV infection, AIDS,

ARC

Zalcitabine

Hoffmann-LaRoche

HIV infection, AIDS,

ARC, with AZT

Zidovudine; AZT

Glaxo Wellcome

HIV infection, AIDS,

ARC, Kaposi's

sarcoma, in

combination with

other therapies

IMMUNOMODULATORS

Drug Name

Manufacturer

Indication

AS-101

Wyeth-Ayerst

AIDS

Bropirimine

Pharmacia Upjohn

Advanced AIDS

Acemannan

Carrington Labs, Inc.

AIDS, ARC

(Irving, TX)

CL246,738

American Cyanamid

AIDS, Kaposi's

Lederle Labs

sarcoma

EL10

Elan Corp, PLC

HIV infection

(Gainesville, GA)

FP-21399

Fuki ImmunoPharm

Blocks HIV fusion

with CD4+ cells

Gamma Interferon

Genentech

ARC, in combination

w/TNF (tumor

necrosis factor)

Granulocyte

Genetics Institute

AIDS

Macrophage Colony

Sandoz

Stimulating Factor

Granulocyte

Hoechst-Roussel

AIDS

Macrophage Colony

Immunex

Stimulating Factor

Granulocyte

Schering-Plough

AIDS,

Macrophage Colony

combination

Stimulating Factor

w/AZT

HIV Core Particle

Rorer

Seropositive HIV

Immunostimulant

IL-2

Cetus

AIDS, in combination

Interleukin-2

w/AZT

JL-2

Hoffman-LaRoche

AIDS, ARC, HIV, in

Interleukin-2

Immunex

combination w/AZT

IL-2

Chiron

AIDS, increase in

Interleukin-2

CD4 cell counts

(aldeslukin)

Immune Globulin

Cutter Biological

Pediatric AIDS, in

Intravenous

(Berkeley, CA)

combination w/AZT

(human)

IMREG-1

Imreg

AIDS, Kaposi's

(New Orleans, LA)

sarcoma, ARC, PGL

IMREG-2

Imreg

AIDS, Kaposi's

(New Orleans, LA)

sarcoma, ARC, PGL

Imuthiol Diethyl

Merieux Institute

AIDS, ARC

Dithio Carbamate

Alpha-2

Schering Plough

Kaposi's sarcoma

Interferon

w/AZT, AIDS

Methionine-

TNI Pharmaceutical

AIDS, ARC

Enkephalin

(Chicago, IL)

MTP-PE

Ciba-Geigy Corp.

Kaposi's sarcoma

Muramyl-Tripeptide

Granulocyte

Amgen

AIDS, in combination

Colony Stimulating

w/AZT

Factor

Remune

Immune Response

Immunotherapeutic

Corp.

rCD4

Genentech

AIDS, ARC

Recombinant

Soluble Human CD4

rCD4-IgG

AIDS, ARC

hybrids

Recombinant

Biogen

AIDS, ARC

Soluble Human CD4

Interferon

Hoffman-LaRoche

Kaposi's sarcoma

Alfa 2a

AIDS, ARC,

in combination

w/AZT

SK & F106528

Smith Kline

HIV infection

Soluble T4

Thymopentin

Immunobiology

HIV infection

Research Institute

(Annandale, NJ)

Tumor Necrosis

Genentech

ARC, in combination

Factor; TNF

w/gamma Interferon

ANTI-INFECTIVES

Drug Name

Manufacturer

Indication

Clindamycin with

Pharmacia Upjohn

PCP

Primaquine

Fluconazole

Pfizer

Cryptococcal

meningitis,

candidiasis

Pastille

Squibb Corp.

Prevention of

Nystatin Pastille

oral candidiasis

Ornidyl

Merrell Dow

PCP

Eflornithine

Pentamidine

LyphoMed

PCP treatment

Isethionate (IM & IV)

(Rosemont, IL)

Trimethoprim

Antibacterial

Trimethoprim/sulfa

Antibacterial

Piritrexim

Burroughs Wellcome

PCP treatment

Pentamidine

Fisons Corporation

PCP prophylaxis

Isethionate for

Inhalation

Spiramycin

Rhone-Poulenc

Cryptosporidial

diarrhea

Intraconazole-

Janssen-Pharm.

Histoplasmosis;

R51211

cryptococcal

Meningitis

Trimetrexate

Warner-Lambert

PCP

Daunorubicin

NeXstar, Sequus

Kaposi's sarcoma

Recombinant Human

Ortho Pharm. Corp.

Severe anemia

Erythropoietin

assoc. with AZT

Therapy

Recombinant Human

Serono

AIDS-related

Growth Hormone

wasting, cachexia

Megestrol Acetate

Bristol-Myers Squibb

Treatment of

Anorexia assoc.

W/AIDS

Testosterone

Alza, Smith Kline

AIDS-related wasting

Total Enteral

Norwich Eaton

Diarrhea and

Nutrition

Pharmaceuticals

malabsorption

Related to AIDS

Additionally, the compounds of the invention herein may be used in combination with another class of agents for treating AIDS which are called HIV entry inhibitors. Examples of such HIV entry inhibitors are discussed in DRUGS OF THE FUTURE 1999, 24(12), pp. 1355-1362; CELL, Vol. 9, pp. 243-246, Oct. 29, 1999; and DRUG DISCOVERY TODAY, Vol. 5, No. 5, May 2000, pp. 183-194.

It will be understood that the scope of combinations of the compounds of this invention with AIDS antivirals, immunomodulators, anti-infectives, HIV entry inhibitors or vaccines is not limited to the list in the above Table, but includes in principle any combination with any pharmaceutical composition useful for the treatment of AIDS.

Preferred combinations are simultaneous or alternating treatments of with a compound of the present invention and an inhibitor of HIV protease and/or a non-nucleoside inhibitor of HIV reverse transcriptase. An optional fourth component in the combination is a nucleoside inhibitor of HIV reverse transcriptase, such as AZT, 3TC, ddC or ddI. A preferred inhibitor of HIV protease is indinavir, which is the sulfate salt of N-(2(R)-hydroxy-1-(S)-indanyl)-2(R)-phenylmethyl-4-(S)-hydroxy-5-(1-(4-(3-pyridyl-methyl)-2(S)-N′-(t-butylcarboxamido)-piperazinyl))-pentaneamide ethanolate, and is synthesized according to U.S. Pat. No. 5,413,999. Indinavir is generally administered at a dosage of 800 mg three times a day. Other preferred protease inhibitors are nelfinavir and ritonavir. Another preferred inhibitor of HIV protease is saquinavir which is administered in a dosage of 600 or 1200 mg tid. Preferred non-nucleoside inhibitors of HIV reverse transcriptase include efavirenz. The preparation of ddC, ddI and AZT are also described in EPO 0,484,071. These combinations may have unexpected effects on limiting the spread and degree of infection of HIV. Preferred combinations include those with the following (1) indinavir with efavirenz, and, optionally, AZT and/or 3TC and/or ddI and/or ddC; (2) indinavir, and any of AZT and/or ddI and/or ddC and/or 3TC, in particular, indinavir and AZT and 3TC; (3) stavudine and 3TC and/or zidovudine; (4) zidovudine and lamivudine and 141W94 and 1592U89; (5) zidovudine and lamivudine.

In such combinations the compound of the present invention and other active agents may be administered separately or in conjunction. In addition, the administration of one element may be prior to, concurrent to, or subsequent to the administration of other agent(s).

Procedures for making compounds of formula I are shown in Schemes 1-13, and further exemplified in Tables 5-8.

Starting indoles 1 (Scheme 1) are known or are readily prepared according to literature procedures, such as those described in Cribble, G. (Ref. 24) or Bantoli et al (Ref. 36). The indoles 1 are treated with oxalyl chloride in either THF (tetrahydrofuran) or ether to afford the desired glyoxyl chlorides 2 according to literature procedures (Lingens, F. et al, Ref. 25). The intermediate glyoxyl chlorides 2 are then coupled with benzoyl piperazine 3 (Desai, M. et al, Ref. 26) under basic conditions to afford 4.

Treatment of indole-3-glyoxyl chloride 2 (Scheme 2) with tert-butyl 1-piperazinecarboxylate 5 affords the coupled product 6. Deprotection of the Boc group of 6 is effected with 20% (trifluoroacetic acid) TFA/CH

2

Cl

2

to yield 7. This product is then coupled with carboxylic acid in the presence of polymer supported 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide (P-EDC) to afford products 8.

For Examples 58-81, piperazine 7 (Scheme 3) was treated with Boc-protected aminobenzoic acid in the presence of EDC to afford 8a. A portion of the resulting product was separated and subjected to TFA in order to remove the Boc group, thus yielding amino derivatives 9.

For Examples 82-89, piperazine 7 (Scheme 4) was treated with acetoxybenzoic acid in the presence of EDC to afford 8b. A portion of the resulting product was separated and subjected to LiOM hydrolysis in order to remove the acetate group, thus yielding hydroxy derivatives 10.

Examples containing substituted piperazines are prepared using the general procedures outlined in Schemes 5-13.

Substituted piperazines are either commercially available from Aldrich, Co. or prepared according to literature procedures (Behun et al, Ref. 31(a), Scheme 5, eq. 01). Hydrogenation of alkyl substituted pyrazines under 40 to 50 psi pressure in ethanol afforded substituted piperazines. When the substituent was an ester or amide, the pyrazine systems could be partially reduced to the tetrahydropyrazine (Rossen et al, Ref. 31(b), Scheme 5, eq. 02). The carbonyl substituted piperazines could be obtained under the same conditions described above by using commercially available dibenzyl piperazines (Scheme 5, eq. 03).

2-Trifluoromethylpiperazine (Jenneskens et al., Ref. 31c) was prepared through a four step route (Scheme 6). Using Lewis acid TiCl

4

, N,N′-dibenzylethylenediamine 11 reacted with trifluoropyruvates 12 to afford hemiacetal 13, which was reduced at room temperature by Et3SiH in CF3COOH to lactam 14. LiAlH

4

treatment then reduced lactam 14 to 1,4-dibenzyl-2-trifluoromethylpiperazine 15. Finally, hydrogenation of compound 15 in HOAc gave the desired product 2-trifluoromethylpiperazine 16.

Mono-benzoylation of symmetric substituted piperazines could be achieved by using one of the following procedures (Scheme 7). (a) Treatment of a solution of piperazine in acetic acid with acetyl chloride afforded the desired mon-benzoylated piperazine (Desai et al. Ref. 26, Scheme 7, eq. 04). (b) Symmetric piperazines were treated with 2 equivalents of n-butyllithium, followed by the addition of benzoyl chloride at room temperature (Wang et al, Ref. 32, Scheme 7, eq. 05).

Mono-benzoylation of unsymmetric substituted piperazines could be achieved by using one of the following procedures (Scheme 8), in which all the methods were exemplified by mono-alkyl substituted piperazines. (a) Unsymmetric piperazines were treated with 2 equivalents of n-butyllithium, followed by the addition of benzoyl chloride at room temperature to afford a mixture of two regioisomers, which could be separated by chromatography (Wang et al, Ref. 32 and 33(b), Scheme 8 eq. 06); (b) Benzoic acid was converted to its pentafluorophenyl ester, and then further reaction with 2-alkylpiperazine to provide the mono-benzoylpiperazines with the benzoyl group at the less hindered nitrogen (Adamczyk et al, Ref. 33(a), Scheme 8, eq. 07); (c) A mixture of piperazine and methyl benzoate was treated with dialkylaluminum chloride in methylene chloride for 2-4 days to yield the mono-benzoylpiperazine with the benzoyl group at the less hindered nitrogen (Scheme 8, eq. 08); (d) Unsymmetric piperazines were treated with 2 equivalents of n-butyllithium, followed by subsequent addition of triethylsilyl chloride and benzoyl chloride in THF at room temperature to afford mono-benzoylpiperazines with the benzoyl group at the more hindered nitrogen (Wang et al, Ref. 33(b), Scheme 8, eq. 09). When the substituent at position 2 was a ester or amide, the mono-benzoylation with benzoyl chloride occurred at the less hindered nitrogen of the piperazine with triethylamine as base in THF (Scheme 8, eq. 10).

In the case of tetrahydropyrazines (Scheme 9, eq. 11), mono-benzoylation occurred at the more hindered nitrogen under the same conditions as those in equation 10 of Scheme 8, in the well precedented manner. (Rossen et al, Ref. 31(b)).

Furthermore, the ester group can be selectively reduced by NaBH

4

in the presence of the benzamide (Masuzawa et al, Ref. 34), which is shown in Scheme 10.

Hydrolysis of Ester Group to Acid:

The ester groups on either the piperazine linkers or on the indole nucleus could be hydrolyzed to the corresponding acid under basic conditions such as K

2

CO

3

(Scheme 11, eq. 13) or NaOMe (Scheme 11, eq. 14) as bases in MeOH and water.

Coupling Reaction:

Reaction of glyoxyl chloride 2 with substituted benzoyl piperazines or tetrahydropyrazines (17) in CH2Cl2 using i-Pr2NEt as base afforded the desired products 18.

In the case of coupling reactions using 3-hydroxylmethylbenzoylpiperazine, the hydroxyl group was temporarily protected as its TMS (trimethylsilyl) ether with BSTFA (N,O-bistrimethylsilyl)fluoroacetamide) (Furber et al, Ref. 35). The unprotected nitrogen atom was then reacted with glyoxyl chlorides 2 to form the desired diamides. During workup, the TMS masking group was removed to give free hydroxylmethylpiperazine diamides 19 (Scheme 13).

Antiviral Activity

The antiviral activity of the compounds of Examples 1-34 was determined in MT-2 cells (a CD4 positive T-lymphocytic cell line) acutely infected by the BRU strain of HIV-1 in the presence of 10 μM compound. The virus yields were quantitated 6 days after infection using a reverse transcriptase assay (Potts, Ref. 27). The anti-viral results are summarized in Table 1, shown below. Cytotoxicity was determined by incubating cells in the presence of serially diluted compound and cell viability determined using an XTT dye reduction assay (Weislow, Ref. 28). The 50% cytotoxicity concentrations of all compounds were significantly higher than 10 μM, indicating that the compounds are relatively non-toxic.

The antiviral activity of the compounds of Examples 35-215 was determined in HeLa CD4 CCR

5

cells infected by single-round infectious HIV-1 reporter virus in the presence of compound at concentrations≦10 μM. The virus infection was quantified 3 days after infection by measuring luciferase expression from integrated viral DNA in the infected cells (Chen et al, Ref. 41). The percent inhibition for each compound was calculated by quantifying the level of luciferase expression in cells infected in the presence of each compound as a percentage of that observed for cells infected in the absence of compound and subtracting such a determined value from 100. Compounds exhibiting anti-viral activity without appreciable toxicity at concentrations ≦10 μM are presented in Tables 1-4 and 9-13.

TABLE 1

%

Example

Inhibition

#

R

1

R

2

R

3

@ 10 uM

1

H

H

H

>98

2

H

H

2,6-Difluoro-

66.8

3

H

H

2,4-Difluoro-

28.5

4

H

H

2-Fluoro-3-

91.2

chloro-

5

H

H

2-Fluoro-

>98

6

H

H

2-Acetoxy-

65.4

7

H

H

2-Hydroxy-

88

8

H

H

3-Chloro-4-

46

fluoro-

9

H

H

3-Fluoro-4-

23.4

methyl-

10

H

H

3,4-Difluoro-

32.7

11

H

H

3-Fluoro-

92.5

12

H

H

3-Bromo-

88.4

13

H

H

4-Hydroxy-

87.7

14

H

H

4-Fluoro-

73.6

15

H

H

4-Methyl-

61.8

16

H

H

4-tertButyl-

47.9

17

H

H

4-Acetoxy-

91.5

18

2-Methyl-

H

H

54.9

19

4-Fluoro-

H

H

>98

20

4-Chloro-

H

H

>98

21

4-Nitro-

H

H

88.3

22

5,6-Diacetoxy-

H

H

34

23

5-Fluoro-

H

H

94.6

24

5-Acetoxy-

H

H

75.8

25

5-Methyl-

H

H

49.5

26

5-Bromo-

H

H

>98

27

5-Chloro-

H

H

95.8

28

6-Fluoro-

H

H

>98

29

6-Chloro-

H

H

95.6

30

6-Methoxy-

H

H

96.4

31

7-Chloro-

H

H

>98

32

7-Carboethoxy-

H

H

>98

33

7-Ethyl-

H

H

>98

34

7-Methyl-

H

H

>98

35

7-Bromo

H

H

>98

36

7-Methoxy

H

H

>98

37

6-Trifluoromethyl

H

H

38

38

7-Fluoro

H

H

>98

39

4,7-Dimethoxy

H

H

>98

40

5,6-Dichloro

H

H

>98

41

4-Bromo

H

H

>98

42

4,6-Difluoro

H

H

>98

43

5-Fluoro-6-chloro

H

H

60

44

5,6-Difluoro

H

H

63

45

4,5,6,7-tetrafluoro

H

H

>98

46

4,7-Difluoro

H

H

>98

47

4-Methoxy

H

H

>98

48

5-Fluoro-7-bromo

H

H

75

49

4-Fluoro-7-methyl

H

H

>98

50

4,6-Difluoro-5-

H

H

97

Bromo

51

4-Fluoro-7-

H

H

>98

trifluoroethoxy

52

4-Methoxy-7-chloro

H

H

>98

53

4-ethoxy

H

H

>98

54

4-methoxy-7-bromo

H

H

>98

55

4-Bromo-7-fluoro

H

H

97

56

4-Fluoro-7-methoxy

H

H

>98

57

4-

H

H

>98

Trifluoromethoxy-

7-bromo

58

4-Fluoro

H

4-NHC(O)OBu-t

95

59

4-Chloro

H

2-

51

NHC(O)OBu-t

60

4-Chloro

H

3-

97

NHC(O)OBu-t

61

4-Chloro

H

4-

93

NHC(O)OBu-t

62

4-Fluoro

H

2-

51

NHC(O)OBu-t

63

4-Fluoro

H

3-

98

NHC(O)OBu-t

64

4-Methoxy

Me

3-

95

NHC(O)OBu-t

65

4-Fluoro

H

2-Amino

84

66

4-Fluoro

H

3-Amino

>98

67

4-Fluoro

H

4-Amino

>98

68

4-Chloro

H

2-Amino

75

69

4-Chloro

H

3-Amino

>98

70

4-Chloro

H

4-Amino

98

71

H

H

2-Amino

36

72

H

H

3-Amino

87

73

4,7-Difluoro

H

3-Amino

>98

74

4,7-Difluoro

Me

3-Amino

>98

75

4,7-Difluoro

Me

4-Amino

>98

76

4-Fluoro

Me

3-Amino

>98

77

4-Methoxy-7-

Me

3-Amino

>98

Chloro

78

4-Methoxy-7-

Me

4-Amino

>98

79

4-Methoxy

Me

3-Amino

>98

80

4-Fluoro-7-

Me

3-Amino

>98

Methoxy

81

4-Fluoro-7-

Me

4-Amino

>98

Methoxy

82

4-Fluoro

H

2-Acetoxy

91

83

4-Fluoro

H

3-Acetoxy

>98

84

4-Fluoro

H

4-Acetoxy

98

85

4-Chloro

H

4-Acetoxy

93

86

4-Chloro

H

3-Acetoxy

>98

87

4-Fluoro

H

2-OH

86

88

4-Fluoro

H

3-OH

>98

89

4-Fluoro

H

4-OH

95

90

4-Fluoro-7-

H

H

>98

carboxaldehyde

TABLE 2

%

Example

Inhibition

#

R

1

W

@ 10 uM

91

4-Fluoro

>98

92

4-Fluoro

94

93

4-Fluoro

>98

94

4-Fluoro

>98

95

4-Fluoro

>98

96

4-Fluoro

>98

97

4-Fluoro

97

98

4-COOMe

84

99

4-Fluoro

89

100

7-COOMe

86

101

4-Fluoro

74

102

7-COOMe

>98

103

7-COOMe

>98

104

7-COOMe

>98

105

7-OMe

>98

106

4,7- Difluoro

>98

107

4,5,6,7- tetrafluoro

>98

108

4,5,6,7- tetrafluoro

>98

109

7-Nitro

>98

110

7-Ethyl

>98

111

7-OMe

>98

112

7-Nitro

84

113

6-Chloro

81

114

5,6- Dichloro

89

115

4-Chloro

79

116

4-Chloro

77

117

5,6- dichloro

89

118

5-Fluoro

69

119

7-Ethyl

72

120

4-Bromo

58

121

7-COOMe

92

122

4-Br

40

123

5-Fluoro

95

124

6-Chloro

>98

125

7-COOMe

>98

126

7-COOMe

>98

127

7-COOMe

95

128

7-COOMe

95

129

7-COOMe

81

130

7-COOMe

77

131

7-COOMe

63

132

7-COOMe

59

133

7-COOMe

92

134

7-COOMe

87

135

4-OMe

>98

136

4-OMe

>98

137

7-COOH

>98

138

4-Fluoro-7- methyl

>98

139

7-COOMe

>98

140

7-Fluoro

>98

141

7-Fluoro

96

142

7-COOMe

>98

143

4-Fluoro

91

144

4-fluoro-7- bromo

>98

145

4-Fluoro-7- COOMe

>98

146

4-Fluoro-7- COOH

>98

147

4-Fluoro-7- OMe

>98

TABLE 3

%

Inhibition

Entry

R

Ar

at 10 μM

148

4-Fluoro

3-Thiophenyl

>98

149

4-Fluoro

1,2,3-Thiadiazolyl

88

150

4-Fluoro

2-(4-Methoxy)-

81

thiophenyl

151

4-Fluoro

2-(5-Methylthio)-

75

thiophenyl

152

4-Fluoro

2-(3-Bromo)-

97

thiophenyl

153

4-Fluoro

2-(5-Bromo)-

>98

thiophenyl

154

4-Fluoro

2-Pyrazinyl

80

155

4-Fluoro

2-(5-Methyl)-

>98

thiophenyl

156

4-Fluoro

2-(5-Chloro)-

>98

thiophenyl

157

4-Fluoro

2-Indolyl

70

158

4-Fluoro

4-(2-Methyl)-

92

thiazolyl

159

4-Fluoro

4-Thiazolyl

>98

160

4-Fluoro

4-Pyridyl

98

161

4-Fluoro

3-(6-Methyl)-

94

pyridyl

162

4-Fluoro

3-Pyridyl

97

163

4-Fluoro

5-Isoxazolyl

90

164

4-Fluoro

2-Furanyl

>98

165

4-Fluoro

3-Pyrazolyl

92

166

4-Fluoro

2-Pyridyl

>98

167

4-Fluoro

3-Furanyl

>98

168

4-Fluoro

2-Thiophenyl

>98

169

4-Fluoro

2-Benzofuranyl

80

170

4-Fluoro

2-(5-Bromo)-

>98

furanyl

171

4-Fluoro

2-(3-Methyl)-

>98

furanyl

172

4-Fluoro

2-(3-Chloro)-

>98

thiophenyl

173

4-Fluoro

3-(5-Chloro-4-

>98

methoxy)-

thiophenyl

174

4-Fluoro

2-(5-Chloro)-

>98

furanyl

175

4-Chloro

3-Thiophenyl

>98

176

4-Chloro

2-[5-(Pyrid-2-yl)]-

86

thiophenyl

177

4-Chloro

2-Thieno[3,2-B]-

>98

thiophenyl

178

4-Chloro

2-(5-Methylthio)-

93

thiophenyl

179

4-Chloro

2-(5-Bromo)-

>98

thiophenyl

180

4-Chloro

2-Pyrazinyl

76

181

4-Chloro

2-Pyridyl

96

182

4-Chloro

2-

89

Benzothiophenyl

183

4-Chloro

2-(5-Chloro)-

>98

thiophenyl

184

4-Chloro

2-(3-Chloro)-

94

thiophenyl

185

4-Chloro

2-Indolyl

85

186

4-Chloro

4-(2-Methyl)-

91

thiazolyl

187

4-Chloro

4-Thiazolyl

98

188

4,7-Difluoro

2-(5-Chloro)-

>98

furanyl

189

4,7-Difluoro

2-(5-Bromo)-

>98

furanyl

190

4,7-Difluoro

2-furanyl

>98

191

4,7-Difluoro

2-Pyridyl

95

192

4,7-Difluoro

2-(3,4-Dichloro)-

>98

furanyl

193

4,7-Difluoro

2-(5-

>98

Trifluoromethyl)-

furanyl

194

4,7-Difluoro

2-(4,5-Dimethyl)-

>98

furanyl

195

4-Fluoro

2-Imidazolyl

71

TABLE 4

%

Inhibition

Entry

R

Ar

@ 10 μM

196

4,7-

2-Pyridyl

>98

difluoro

197

4-Fluoro-7-

2-Pyridyl

93

methyl

198

4,7-

2-(5-Bromo)-

>98

Difluoro

furanyl

199

4,7-

2-(5-Bromo)-

>98

Dimethoxy

furanyl

200

7-COOMe

2-(5-Bromo)-

>98

furanyl

201

4,7-

2-Pyridyl

>98

Difluoro

202

4-Fluoro

2-Pyridyl

>98

203

4-Chloro

2-Pyridyl

80

204

4-Bromo

2-Pyridyl

50

205

5-Fluoro

2-Pyridyl

61

206

6-Chloro

2-Pyridyl

88

207

7-Fluoro

2-Pyridyl

>98

208

7-Methoxy

2-Pyridyl

>98

209

7-Methyl

2-Pyridyl

>98

210

7-Ethyl

2-Pyridyl

88

211

4-methoxy-

2-Pyridyl

>98

7-chloro

212

7-cyano

2-Pyridyl

>98

213

4-Methoxy

2-Pyridyl

>98

214

4-

2-Pyridyl

>98

Methoxy-

7-Bromo

215

4-Fluoro-7-

2-Pyridyl

>98

Methoxy

EXPERIMENTAL PROCEDURES

Biology

Abbreviations

“μM” means micromolar;

“μci” means microcurie;

“ml” means milliliter;

“μl” means microliter;

“μg” means microgram;

“M” means molar;

“μm” means micromolar;

“mM” means millimolar;

“a” refers to percent inhibition data as representing the mean values of at least two experiments with duplicate determinations in each experiment.

“RT” refers to reverse transcriptase.

The materials and experimental procedures used to obtain the anti-viral results for Examples 1-34 are described below.

Cells—MT-2 T cell lines propagated in Roswell Park Memorial Institute (RPMI) 1640 medium (Life Technologies, Gaithersburg, Md.) containing 10% fetal Bovine serum (FBS, Sigma, St. Louis, Mo.).

Virus—The laboratory HIV-1 strain BRU was titrated using an infectivity assay (johnson, V. A. and R. E. Byrington, 1990).

Experiment

1. MT-2 cells (Harada, et al, Ref. 30) were infected by HIV-1 BRU at a multiplicity of infection (MOI) of 0.005 in RPMI 1640 medium containing 10% fetal Bovine serum at a concentration of 1×10

5

cells/ml.

2. Compound was added to 100 μl RPMI 1640 media containing 10% fetal Bovine serum per well in a 96 well plate at a concentration of 20 μM.

3. 100 μl of 1×10

5

/ml infected MT-2 cells were added to each well in such plates, resulting in a final cell concentration of 5×10

4

cells/ml and a final compound concentration of 10 μM.

4. Samples were incubated at 37° C. and harvested 6 days after infection.

5. HIV-1 replication was quantified by measuring the HIV-1 reverse transcriptase (RT) activity present in cell-free supernatants (Potts, et al, Ref. 27). For each sample, 20 μl of cell-free supernatant was added to 40 μl RT cocktail [42 μM Tris(hydroxymethyl) aminomethane pH 7.8 (Sigma, St. Louis, Mo.), 63 μM potassium chloride (Mallinckrodt, Paris Ky.), 2 μM dithiothreitol (Sigma, St. Louis, Mo.), 4 μM magnesium chloride (Mallinckrodt, Paris Ky.), 4 μg/ml polyadenylic acid (Pharmacia, Piscataway, N.J.), 1.3 μg/ml oligonucleotide deoxythymidinel 2-18 (Pharmacia, Piscataway, N.J.), 0.04% (octylphenoxy)-polyethoxyethanol (Nonidet P40, Sigma, St. Louis, Mo.), and 17 μCi/ml 3H-deoxythymidine 5′-triphosphate (NEN, Boston, Mass.)]. Assays were incubated for 1 hour at 37° C. and then 1 μl portions of each reaction were spotted on diethylaminoethyl cellulose (DE-81) filter paper (Whatman, Hillsboro, Oreg.), allowed to dry, washed four times with 0.3 M sodium chloride (Fisher Scientific, Pittsburgh, Pa.), 30 mM sodium citrate pH 7.0 (Sigma, St. Louis, Mo.), followed by two washes in 95% ethanol. Bound radioactivity was quantified by scintillation counting.

6. The percent inhibition for each compound was calculated by quantifying the level of HIV-1 replication in the presence of each compound as a percentage of the no compound control and subtracting such a determined value from 100.

7. To determine the cytotoxicity of compounds, uninfected cells were incubated with a series of concentrations of each compound for 3-6 days. Cell viability was determined by the XTT {2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide} dye reduction method (Weislow et al, Ref. 28). The percentage of living cells in compound containing wells compared to untreated controls was determined. The 50% cytotoxic concentration was calculated as the concentration of drug that decreased the percentage of living cells to 50% of those in untreated cells.

The materials and methods for determination of anti-viral activity for examples 35-215 are described below:

Cells:

Virus production—Human embryonic Kidney cell line, 293, propagated in Dulbecco's Modified Eagle Medium (Life Technologies, Gaithersburg, Md.) containing 10% fetal Bovine serum (FBS, Sigma, St. Louis, Mo.).

Virus infection—Human epithelial cell line, HeLa, expressing the HIV-1 receptors CD4 and CCR5 was propagated in Dulbecco's Modified Eagle Medium (Life Technologies, Gaithersburg, Md.) containing 10% fetal Bovine serum (FBS, Sigma, St. Louis, Mo.) and supplemented with 0.2 mg/ml Geneticin (Life Technologies, Gaithersburg, Md.) and 0.4 mg/ml Zeocin (Invitrogen, Carlsbad, Calif.).

Virus-Replication defective reporter virus was produced by co-transfecting human embryonic Kidney 293 cells with an HIV-1 envelope DNA expression vector and a proviral cDNA containing an envelope deletion mutation and the luciferase reporter gene inserted in place of HIV-1 nef sequences (Chen, 1994). Transfections were performed using lipofectAMINE PLUS reagent as described by the manufacturer (Life Technologies, Gaithersburg, Md.).

Experiment

1. Compound was added to HeLa CD4 CCR

5

cells plated in 96 well plates at a cell density of 5×10

4

cells per well in 100 ul Dulbecco's Modified Eagle Medium containing 10% fetal Bovine serum at a concentration of <20 uM.

2. 100 ul of replication defective reporter virus in Dulbecco's Modified Eagle Medium was then added to the plated cells and compound at a multiplicity of infection (MOI) of 0.01, resulting in a final volume of 200 ul per well and a final compound concentration of <10 uM.

3. Samples were harvested 72 hours after infection.

4. Viral infection was monitored by measuring luciferase expression from viral DNA in the infected cells using a luciferase reporter gene assay kit (Roche Molecular Biochemicals, Indianapolis, Ind.). Infected cell supernatants were removed and 50 ul of Dulbecco's Modified Eagle Medium (without phenol red) and 50 ul of luciferase assay reagent reconstituted as described by the manufacturer (Roche Molecular Biochemicals, Indianapolis, Ind.) was added per well. Luciferase activity was then quantified by measuring luminescence using a Wallac microbeta scintillation counter.

5. The percent inhibition for each compound was calculated by quantifying the level of luciferase expression in cells infected in the presence of each compound as a percentage of that observed for cells infected in the absence of compound and subtracting such a determined value from 100.

References

Chen, B. K., Saksela, K., Andino, R., and D. Baltimore. 1994. Distinct modes of human immunodeficiency type 1 proviral latency revealed by superinfection of nonproductively infected celllines with recombinant luciferase-encoding viruses. J. Virol. 68:654-660 (Ref. 37).

Chemistry

General: Unless otherwise noted, solvents and reagents were used directly as obtained from commercial sources, and reactions were performed under the nitrogen atmosphere. Flash chromatography was conducted on Silica gel 60 (0.040-0.063 particle size; EM Science supply).

1

H NMR spectra were recorded at 500 MHz, unless otherwise noted, and the chemical shifts are reported relative to residual solvent signals. The following standard acronyms were employed to describe the multiplicity patterns: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), b (broad), app (apparent). The coupling constant (J) is in hertz.

All Liquid Chromatography (LC) data were recorded on a Shimadzu LC-10AS liquid chromotograph using a SPD-10AV UV-Vis detector and Mass Spectrometry (MS) data were determined with a Micromass Platform for LC in electrospray mode.

LC/MS Method (i.e., Compound Identification)

Unless otherwise noted, all compounds were analyzed using the following conditions:

Column: YMC ODS S7 3.0×50 mm column

Gradient: 100% Solvent A/0% Solvent B to 0% Solvent A/100% Solvent B

Gradient time: 2 minutes

Hold time: 1 minute

Flow rate: 5 mL/min

Detector Wavelength: 220 nm

Solvent A: 10% MeOH/90% H

2

O/0.1% Trifluoroacetic Acid

Solvent B: 10% H

2

O/90% MEOH/0.1% Trifluoroacetic Acid

When noted, the following conditions were used for HPLC analysis:

Method A: Column YMC ODS-A C18 S7 3.0×50 min

Start%B=0/Finish%B=100

Method B: Column YMC ODS-A C18 S7 3.0×50 min

Start%B=30/Finish%B=100

Method C: Column PHX-LUNA C18 4.6×30 mm

Start%B=0/Finish%B=100

Compounds purified by preparative HPLC were diluted in methanol (1.2 mL) and purified using the following methods on a Shimadzu LC-10A automated preparative HPLC system.

Preparative HPLC Method (i.e., Compound Purification)

Purification Method: Initial gradient (40% B, 60% A) ramp to final gradient (100% B, 0% A) over 20 minutes, hold for 3 minutes (100% B, 0% A)

Solvent A: 10% MeOH/90% H

2

O/0.1% Trifluoroacetic Acid

Solvent B: 10% H

2

O/90% MeOH/0.1% Trifluoroacetic Acid

Column: YMC C18 S5 20×100 mm column

Detector Wavelength: 220 nm

Indoles were either commercially available, or were prepared using known chemistry such as the method of Bartoli (Ref. 36) or as described by Gribble (Ref. 24)

Representative indole syntheses are shown below.

Preparation of 4-Fluoro-7-methyl Indole

Step A

A flame dried 50 ml three neck flask was charged with BCl3 (44 mmol, 44 ml, 1 M in benzene) and 10 mL dry benzene at rt. under N2. The mixture was cooled down to 0° C. followed by dropwise addition of 5-fluoro-2-methylaniline(5 g, 40 mmol) in 10 ml dry benzene over 10 min, chloroacetonitrile (2.18 g, 48 mmol) over 2 min and AlCl3 in one portion. After stirring at 0° C. for 5 min, the ice-bath was removed and the mixture was refluxed for 6 hr under N2. The resulted mixture was cooled down to rt. and poured into EtOAc/1N HCl (300 mL, 50:50 v:v with ice). After separation, the aqueous phase was extracted with EtOAc (2×100 mL). The combined organic layers were washed with water (100 mL), brine (2×100 mL), and dried with MgSO4. The solvent was removed in vacuo, and the crude intermediate was used directly in next step without further purification.

Step B

The above residue was dissolved in 100 mL ETOH. Then the mixture was cooled down to 0° C., followed by dropwise addition of NaBH4 in 2 ml H2O. After stirring at 0° C. for 1 hr, the reaction was quenched with H2O (10 ml). The solvent was removed in vacuo and the residue was dissolved in EtOAc (150 ml), and washed with brine (2×50 ml). The organic layer was dried with MgSO4, the solvent was removed, and the expected reduced intermediate was used directly in next cyclization step.

Step C:

The above intermediate, a yellow oil, was dissolved in 100 ml EtOH, followed by addition of K2CO3 (11.0 g, 80 mmol). The mixture was refluxed under N2 for 2 hr, and cooled down to rt. The solids were removed by filtering through celite, and the resulted solution was concentrated in vacuo. The residue was dissolved in EtOAc (200 mL), washed with brine (2×50 ml) and dried over MgSO4. The solvent was removed in vacuo and gave a brown oil which was purified by flash chromotography (12% EtOAc in hexanes), to yield 2.3 g (39% overall yield) of pure product. M+H, 150.0; Retention time, 1.297 min.

Synthesis of 4-Ethoxyindole

An oven dried two neck flask was charged with 5 ml DMF and NaH (66 mg, 60% in oil, 1.65 mmol). The mixture was cooled down to 0° C., followed by dropwise addition of 4-hydroxy indole (200 mg, 1.5 mmol) in 5 ml DMF over 10 second. After stirring for 30 min under N2, bromoethane in 2 ml DMF was added dropwise, and the reaction was allowed to warm to rt. with continued stirring for 2 hr. Removal of the solvent in vacuo, followed by aqueous work up afforded crude 4-ethoxyindole which was purified by prep. HPLC to afford 201 mg (83%) of pure 4-ethxyindole; HPLC retention time, 1.190 min.

Synthesis of 4-Fluoro-7-carbomethoxy Indole

Step A:

A mixture of 4-fluoro-7-bromoindole (600 mg, 2.8 mmol) and CuCN (1.004 g, 11.2 mmol) in DMF (4 ml) was refluxed for 16 hours. After cooling to room temperature, the reaction mixture was poured into a solution of ammonia in MeOH (30 ml, sat.) and the residue removed by filtration. The filtrate was added to a mixture of water (20 ml)/ammonia (20 ml, sat. aq.) and extracted with EtOAc/Ether (1/1) until TLC analysis showed no product in the aqueous phase. The combined organic extracts were washed with brine (2×200 ml) and water (200 ml), dried (MgSO

4

); evaporation in zacuo gave 4-fluoro-7-cyanoindole as a tan yellow solid (310 mg, 69%).

Step B

To a solution of KOH (13.04 g, 0.232 mol) in 14% H

2

O/EtOH (50 ml) was added 4-fluoro-7-cyanoindole (900 mg, 5.60 mmol). The resulting mixture was refluxed for 12 hours, slowly cooled to room temperature, and concentrated in vacuo to about 30 ml. The residue was acidified to pH 2 with HCl (˜5.5 N aq.). The precipitate was filtered, washed with excess of water, and dried under high vacuum to afford 4-fluoro-7-carboxyindole as a white solid (100% conversion). The material was used without further purification.

Step C

To a suspension of 4-fluoro-7-carboxyindole in a mixture of MeOH (18 ml)/PhH (62 ml) was added (trimethylsilyl)diazomethane (8.8 ml, 17.6 mmol, 2 M in hexane). The resulting mixture was stirred at room temperature for 30 min., quenched with excess acetic acid and evaporated in vacuo. The crude oily material was purified by flash chromatography using a gradient elution (Hexane to 10% EtOAc/Hexane) to afford methyl (4-fluoro)indole-7-carboxylate as a white solid (1.04 g, 83% two steps)

Preparation of 4-Fluoroindole-7-carboxaldehyde

To a solution of 4-fluoro-7-bromoindole (1.0 g, 4.7 mmol) in THF (5 mL) at −78° C. was added n-BuLi (5.6 mL, 2.5M in hexanes) dropwise. The mixture was stirred for 15 min at −78° C., was allowed to warm to 5° C. for 30 min and was then re-cooled to −78° C. DMF (1.8 mL) was then added and the mixture was allowed to warm to room temperature slowly. The reaction was quenched with water and was extracted with ether. The organic phase was dried over MgSO4, filtered and concentrated to afford 4-fluoroindole-7-carboxaldehyde.

General Procedure for Preparation of Examples 1-17 in Table 5

Step A

To commercially available indole-3-glyoxylyl chloride I (3 gram, 14.45 mmol) in CH

2

Cl

2

at room temperature was added tert-butyl 1-piperazinecarboxylate (2.7 gram, 14.45 mmol) and diisopropylethylamine (2.76 ml, 15.9 mmol). The light-brown color solution was stirred for 2 hr at room temperature after which time LC/MS analysis indicated the completion of the reaction. The solvent was removed in vacuo and the resulting residue was diluted with ethyl acetate (250 ml) and diethylether (250 ml). The organic solution was then washed with water (100 ml×3) and brine (50 ml), dried over MgSO

4

, filtered and concentrated. To the light-yellow solid was then added 30 ml of 20% trifluoroacetic acid in CH

2

Cl

2

. The solution was concentrated and the light-brown solid was dried in vacuo to give 3.5 g (95%) of product II. LC/MS analysis indicated this product was 100% pure and it was used for the next reaction without further purification.

Step B

To piperazine indole-3-glyoxylamide II (0.03 mmol) was added resin-bound 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (P-EDC) (0.21 mmol) and carboxylic acid (RCOOH) (0.06 mmol) in dichloroethane (DCE) (1 mL) or DMF (dimethylformamide) (1 mL) in cases where the carboxylic acids are not soluble in DCE. The reaction was shaken for 12 hr at room temperature. The product III was filtered and concentrated. Products with purity less than 70% were diluted in methanol and purified using a Shimadzu automated preparative HPLC System.

2) General Procedure for Preparation of Examples 18-56 in Table 5

Step A.

To a solution of substituted indole IV (1 eq) in dry Et

2

O was dropwise added oxalylchloride (1.2 eq) at 0° C. After 5 min., the reaction mixture was warmed to room temperature, or heated to 35° C. overnight if necessary. The intermediate substituted-indole-3-glyoxylyl chloride V, 10 which was formed as a solid, was filtered and washed with dry ether (2×1 ml) to remove excessive oxalyl chloride. The product was then dried under vacuum to give desired glyoxyl chlorides V.

In cases where reaction in Et

2

O was unsuccessful, the following procedure was adopted: To a solution of substituted indole IV (1 eq) in dry THF (tetrahydrofuran) solvent was dropwise added oxalyl chloride (1.2 eq) at 0° C. After 5 min., the reaction was warmed to room temperature, or heated to ˜70° C. under nitrogen if necessary. After concentration in vacuo, the resulting crude intermediate V was submitted to next step without further treatment.

Step B

To a solution of indole glyoxyl chloride V (1 eq) in dry THF was added benzoylpiperazine (1 eq) at room temperature. Then the mixture was cooled down to 0° C., followed by dropwise addition of diisopropylamine (1.3 eq). After 5 min., the reaction mixture was warmed to room temperature and was shaken for 3 hr. The resulting crude products VI were purified by preparative HPLC and characterized as shown in Table 5.

General Procedure for Preparation of Examples 58-63 in Table 5

Step A

To glyoxyl chloride V (1 equiv.) in CH

2

Cl

2

at room temperature was added tert-butyl 1-piperazinecarboxylate (1 equiv) and diisopropylethylamine (1.2 equiv). The solution was stirred for 2 hr at room temperature after which time LC/MS analysis indicated the completion of the reaction. The solvent was removed in vacuo and the resulting residue was diluted with ethyl acetate and diethylether. The organic solution was then washed with water (100 ml×3) and brine (50 ml), dried over MgSO

4

, filtered and concentrated. To the solid was then added 30 ml of 20% trifluoroacetic acid in CH

2

Cl

2

. The solution was concentrated and the light-brown solid was dried in vacuo to give glyoxamide VII.

Step B

To piperazine glyoxamide VII (0.1 mmol, 1 eq) in DMF (1 mL) at room temperature was added EDC (1.5 eq) and Boc-aminobenzoic acid (1.5 eq). The reaction mixture was stirred at room temperature for 16 hours. The crude product was then purified by preparative HPLC to afford product VIII.

General Procedure for Preparation of Examples 65-73 in Table 5

To Boc-protected derivative VIII (0.03 mmol) was added 50% TFA/CH

2

Cl

2

(1.5 mL). The reaction mixture was stirred at room temperature for 16 hours. The product was then concentrated in vacuo to afford product IX as its TFA salt. The purity of IX was sufficient that no further purification was necessary.

General Procedure for Preparation of Examples 82-86 in Table 5

To piperazine glyoxamide VII (0.1 mmol, 1 eq) in DMF (1 mL) at room temperature was added EDC (1.5 eq) and acetoxybenzoic acid (1.5 eq). The reaction mixture was stirred at room temperature for 16 hours. The crude product was then purified by preparative HPLC to afford product X.

General Procedure for Preparation of Examples 87-89 in Table 5

To acetate-protected derivative X (0.03 mmol, 1 eq) was added aqueous LiOH (3 eq) in THF/MeOH (1.5 mL, 1:1). The reaction mixture was stirred at room temperature for 16 hours. The crude product was then purified by preparative HPLC to afford product XI.

General Procedure for Preparation of Examples 64 and 74-81 in Table 5

Step A

To a solution of Boc-protected amino benzoic acid (5 mmol) in DMF (10 mL) at room temperature was added pentafluorophenol (5 mmol) followed by EDC (5 mmol). The reaction mixture was stirred at room temperature for 3 h. The crude product was diluted with CH2Cl2 and was washed with water, 0.1 M HCl and brine. The organic phase was dried over MgSO4, filetered and concentrated. The pentafluorophenyl ester XII was used in the following reaction without further purification.

Step B

To a stirred solution (R)-2-methylpiperazine in DMF (15 mL) at room temperature was added a solution of pentafluorophenyl ester XII in DMF (2 mL) dropwise. The reaction mixture was stirred at room temperature for 16 hours. The crude product was diluted with CH2Cl2 and was washed with Na2CO3 (sat) and brine. The organic phase was dried over MgSO4, filtered and concentrated. The crude product was purified by flash chromatography (50% EtOAc/Hexane—10% MeOH/EtOAc) to afford product XIII.

Step C

To indole-3-glyoxyl chloride V (1 eq.) in CH2Cl2 was added acylpiperazine XII followed by i-Pr

2

NEt (3 eq.). The reaction mixture was stirred at room temperature for 5 hours, was then diluted with methanol and product XIV was purified by preparative HPLC.

Step D

To Boc-protected derivative XIV (˜0.03 mmol) was added 50% TFA/CH

2

Cl

2

(1.5 mL). The reaction mixture was stirred at room temperature for 16 hours. The product was then concentrated in vacuo to afford product XV. The purity of XV was sufficient that no further purification was necessary.

Procedure for the Synthesis of Examples 57 and 90 in Table 5.

Step A

To substituted indole IV (1 equiv) in CH

2

Cl

2

at 0° C. was added ethyl chloroxalate (2 equiv) dropwise followed by addition of AlCl

3

(2 equiv). The reaction was stirred at 0° C. and was then allowed to warm to room temperature overnight. The reaction was quenched by dropwise addition of HCl (1N). The crude material was extracted with EtOAc and was washed with water, dried over MgSO4, filtered and concentrated. The crude product was then recrystallised from EtOAc/Hexanes to afford ester XVI.

Step B

To ester XVI (1 equiv) in EtOH was added NaOH (2.5 equiv, 10N) dropwise. The reaction mixture was stirred at rt for 30 min and was then heated to 45° C. for an additional 90 min. The product was concentrated in vacuo. The resulting residue was partitioned between EtOAc and water. The organic layer was separated and was washed with water, dried over MgSO4, filtered and concentrated to afford acid XVII.

Step C

To acid XVII (1 equiv) in DMF was added benzoyl pierazine (1.2 equiv) followed by DEPBT (1.2 equiv) and i-Pr2NEt (2 equiv). The reaction mixture was stirred at room temperature for 2 h. It was then diluted with EtOAc, was washed with water and brine, dried over MgSO4, filtered and concentrated. The crude product was then purified by flash chromatography (EtOAc/MeOH, 95:5) to afford the desired product XVIII.

TABLE 5

HPLC

MS

Retention

Data

Example

Time

(M +

#

R

1

R

2

R

3

(min)

H)

+

1

H

H

H

1.13

362

2

H

H

2,6-

1.36

398

Difluoro-

3

H

H

2,4-

1.43

398

Difluoro

4

H

H

2-Fluoro-3-

1.54

415

chloro-

5

H

H

2-Fluoro-

1.42

380

6

H

H

2-Acetoxy-

1.63

378

7

H

H

2-Hydroxy-

1.25

378

8

H

H

3-Chloro-

1.56

415

4-fluoro-

9

H

H

3-Fluoro-4-

1.54

394

methyl-

10

H

H

3,4-

1.45

398

Difluoro

11

H

H

3-Fluoro-

1.44

380

12

H

H

3-Bromo-

1.56

442

13

H

H

4-Hydroxy-

1.17

378

14

H

H

4-Fluoro-

1.43

380

15

H

H

4-Methyl-

1.34

376

16

H

H

4-tertButyl-

1.76

419

17

H

H

4-Acetoxy-

1.58

420

18

2-Methyl-

H

H

1.86

376

19

4-Fluoro-

H

H

1.21

380

20

4-Chloro-

H

H

1.35

397

21

4-Nitro-

H

H

1.16

407

22

5,6-Diacetoxy-

H

H

1.78

478

23

5-Fluoro-

H

H

1.69

380

24

5-Acetoxy-

H

H

1.76

420

25

5-Methyl-

H

H

1.37

376

26

5-Bromo-

H

H

1.51

442

27

5-Chloro-

H

H

1.58

396

28

6-Fluoro-

H

H

1.80

380

29

6-Chloro-

H

H

2.01

396

30

6-Methoxy-

H

H

1.28

392

31

7-Chloro-

H

H

1.50

397

32

7-Carboethoxy-

H

H

1.41

380

33

7-Ethyl-

H

H

1.57

390

34

7-Methyl-

H

H

1.47

376

35

7-Bromo

H

H

1.54

442

36

7-Methoxy

H

H

1.37

392

37

6-Trifluoromethyl

H

H

ND

430

38

7-Fluoro

H

H

1.59

380

39

4,7-Dimethoxy

H

H

1.30

422

40

5,6-Dichloro

H

H

1.86

432

41

4-Bromo

H

H

1.62

440

42

4,6-Difluoro

H

H

1.44

398

43

5-Fluoro-6-chloro

H

H

1.52

414

44

5,6-Difluoro

H

H

ND

398

45

4,5,6,7-tetrafluoro

H

H

1.54

434

46

4,7-Difluoro

H

H

1.30

398

47

4-Methoxy

H

H

1.23

392

48

5-Fluoro-7-bromo

H

H

ND

ND

49

4-Fluoro-7-methyl

H

H

1.34

394

50

4,6-Difluoro-5-

H

H

1.63

477

Bromo

51

4-Fluoro-7-

H

H

1.44

478

trifluoroethoxy

52

4-Methoxy-7-

H

H

1.36

426

chloro

53

4-ethoxy

H

H

1.28

406

54

4-methoxy-7-

H

H

1.35

471

bromo

55

4-Bromo-7-fluoro

H

H

1.45

458

56

4-Fluoro-7-

H

H

1.29

410

methoxy

57

4-

H

H

ND

525

Trifluoromethoxy-

7-bromo

58

4-Fluoro

H

4-

1.43

495

NHC(O)O

Bu-t

59

4-Chloro

H

2-

1.56

511

NHC(O)O

Bu-t

60

4-Chloro

H

3-

1.55

511

NHC(O)O

Bu-t

61

4-Chloro

H

4-

1.54

511

NHC(O)O

Bu-t

62

4-Fluoro

H

2-

1.42

495

NHC(O)O

Bu-t

63

4-Fluoro

H

3-

1.44

495

NHC(O)O

Bu-t

64

4-Methoxy

Me

3-

1.46

522

NHC(O)O

Bu-t

65

4-Fluoro

H

2-Amino

1.07

395

66

4-Fluoro

H

3-Amino

0.91

395

67

4-Fluoro

H

4-Amino

0.86

395

68

4-Chloro

H

2-Amino

1.21

411

69

4-Chloro

H

3-Amino

1.06

411

70

4-Chloro

H

4-Amino

1.03

411

71

H

H

2-Amino

1.12

377

72

H

H

3-Amino

0.93

377

73

4,7-Difluoro

H

3-Amino

0.98

413

74

4,7-Difluoro

Me

3-Amino

1.02

427

75

4,7-Difluoro

Me

4-Amino

1.01

427

76

4-Fluoro

Me

3-Amino

0.97

409

77

4-Methoxy-7-

Me

3-Amino

1.11

455

Chloro

78

4-Methoxy-7-

Me

4-Amino

1.11

455

Chloro

79

4-Methoxy

Me

3-Amino

0.93

421

80

4-Fluoro-7-

Me

3-Amino

1.05

439

Methoxy

81

4-Fluoro-7-

Me

4-Amino

1.04

439

Methoxy

82

4-Fluoro

H

2-Acetoxy

1.18

438

83

4-Fluoro

H

3-Acetoxy

1.17

438

84

4-Fluoro

H

4-Acetoxy

1.16

438

85

4-Chloro

H

4-Acetoxy

1.28

454

86

4-Chloro

H

3-Acetoxy

1.29

454

87

4-Fluoro

H

2-OH

1.17

396

88

4-Fluoro

H

3-OH

1.13

396

89

4-Fluoro

H

4-OH

1.09

396

90

4-Fluoro-7-

H

H

0.79

408

carboxaldehyde

(M + H)

+

refers to the molecular ion peak in positive ionization mode. ND means not determined.

Procedure for Synthesis of Compounds in Table 6

A. Preparation of Substituted Piperazines:

Preparation of 2-Alkylpiperazines

5 g of 2-alkyl pyrazine (46.3 mmol, from Pyrazine Specialties, Inc.) was dissolved in 200 ml of 95% ethanol with 500 mg 10% palladium on active carbon. The reaction mixture was hydrogenated under pressure (40-50 psi) for 2 days. The solid was filtered and removed. The filtrate was concentrated to afford 2-alkyl piperazine, which did not require further purification.

2-ethylpiperazine XIX:

1

H NMR (500 MHz, CD

3

OD) δ 2.89 (t, J=15.05 Hz, 1H), 2.85 (d, J=15.11 Hz, 2H), 2.75 (t, J=11.80 Hz, 1H), 2.65 (t, J=11.90 Hz, 1H), 2.48 (m, 1H), 2.28 (t, J=6.12 Hz, 1H), 1.35 (m, 2H), 0.93 (t, J=7.55 Hz, 3H).

2-propylpiperazine XX:

1

H NMR (300 MHz, CDCl

3

) δ 3.00-2.60 (m, 6H), 2.65 (t, J=10.20 Hz, 1H), 1.70 (m, 2H), 1.30 (m, 2H), 0.92 (t, J=6.9 Hz, 3H).

2-iso-propylpiperazine XXI:

1

H NMR (300 MHz, CDCl

3

) δ 3.03-2.30 (m, 7H), 1.50 (m, 1H), 0.91 (dd, J=6.60 & 6.60 Hz, 3H).

2-iso-butylpiperazine XXII:

1

H NMR (500 MHz, CD

3

OD) δ 3.00-2.62 (m, 6H), 2.28 (t, J=10.55 Hz, 1H), 1.68 (m, 1H), 1.38 (m, 2H), 0.92 (dd, J=6.65 & 6.55 Hz, 3H).

2-tert-butylpiperaine XXIII:

1

H NMR (500 MHz, CD

3

OD) δ 2.96 (d, J=11.85 Hz, 2H), 2.80 (d, J=12.05 Hz, 1H), 2.74 (t, J=11.75 Hz, 1H), 2.63 (t, J=11.95 Hz, 1H), 2.41 (t, J=11.85 Hz, 1H), 2.31 (d, J=13.91 Hz, 1H), 0.92 (s, 9H).

2-pentylpiperazine XXIV::

1

H NMR (500 MHz, CD

3

OD) δ 2.89 (m, 2H), 2.83 (d, J=11.95 Hz, 1H), 2.75 (t, J=11.80 Hz, 1H), 2.65 (t, J=11.85 Hz, 1H), 2.56 (m, 1H), 2.28 (t, J=12.3 Hz, 1H), 1.35 (m, 8H), 0.90 (t, J=7.15 Hz, 3H).

Preparation of 2-Methoxycarbonyltetrahydropyrazine XXV

5 g of pyrazine carboxylic acid methyl ester (36.2 mmol, from Lancaster, Inc.) was dissolved in 200 ml of 95% ethanol with 500mg 10% palladium on active carbon. The reaction mixture was hydrogenated under pressure (40-50 psi) for 2 days. The solid was filtered and removed. The filtrate was concentrated to afford methoxycarbonyltetrahydropyrazine XXV, which was sufficiently pure enough for subsequent reactions.

2-Methoxycarbonyltetrahydropyrazine XXV:

1

H NMR (300 MHz, CD

3

OD) δ 7.10(s, 1H), 4.84 (b, 2H), 3.66 (s, 3H), 3.29 (t, J=6.0 Hz, 2H), 3.08 (t, J=6.0 Hz, 2H);

13

C NMR (75 MHz, CD

3

OD) δ 166.1, 130.8, 105.4, 48.4, 40.6, 40.0; MS m/z: (M+H)

+

calcd for C

6

H

11

N

2

O

2

: 143.08, found 143.09. HPLC retention time 0.11 (Method C).

Preparation of 2-Ethoxycarbonylpiperazine XXVI

5 g of N,N′-dibenzylpiperazine carbhoxylic acid ethyl ester (14.8 mmol, from Maybridge Chemical Company Ltd.) was dissolved in 200 ml of 95% ethanol with 500 mg 10% palladium on active carbon. The reaction mixture was hydrogenated under pressure (40-50 psi) for 2 days. The solid was filtered and removed. The filtrate was concentrated to afford 2-ethoxycarbonylpiperazine XXVI, which was sufficiently pure for subsequent reactions.

2-Ethoxycarbonylpiperazine XXVI:

1

H NMR (300 MHz, CD

3

OD) δ 4.20 (q, J=7.20 Hz, 2H), 3.46-2.60 (m, 7H), 1.27 (t, J=6.9 Hz, 3H.).

Preparation of 2-Trifluoromethylpiperazine XXVII

Step 1:

To a solution of N,N′-dibenzylethylenediamine (1.51 ml, 6.41 mmol), methyl 3,3,3-trifluoro-2-oxopropanate (1.0 g, 6.41 mmol) and triethylamine (1.78 ml, 12.8 mmol) in dichloromethane (100 ml) was added via a syringe titanium chloride (1M in CH

2

Cl

2

, 3.21 ml, 3.21 mmol). The reaction was stirred for 8 hours and the solvents were removed in vacuo. The residue was carried to the next step without further purification.

Step 2:

The crude product (200 mg, <0.55 mmol) from the previous step was dissolved in TFA (5 ml). An excess of triethylsilane (0.88 ml, 5.5 mmol) was then added. After 30 minutes, TFA was removed under vaccum and the residue was carried to the next step without further purification.

Step 3:

The crude product (<0.55 mmol) from step 2 was suspended in ether. LiAlH

4

(1M in THF, 0.55 ml, 0.55 mmol) was then added at room temperature. After stirring for 8 hours, the reaction was quenched with saturated NaHCO

3

solution. The aqueous layer was extracted with EtOAc. Organic layers were combined, dried over MgSO

4

and concentrated to give a residue, which was carried to the next step without purification.

Step 4:

The crude product from the step 3 was dissolved in HOAc (20 ml) with 10 mg 10% palladium on active carbon. The reaction mixture was hydrogenated under pressure (40-50 psi) for 8 hours. The solid was filtered and removed. The filtrate was concentrated to afford 2-trifluoromethylpiperazine XXVII as a HOAc salt, which was pure enough for the further reactions.

2-Trifluoromethylpiperazine XXVII as its HOAc (2 equivalents) salt:

1

H NMR (300 MHz, CD

3

OD) δ 3.80-2.80 (m, 7H), 1.95 (s, 6H);

13

C NMR (75 MHz, CD

3

OD) δ 174.5, 53.8, 53.3, 42.7, 41.3, 40.8, 19.8; HRMS m/z: (M+H)

+

calcd for C

5

H

10

F

3

N

2

: 155.0796, found 155.0801.

B. Mono-benzoylation of Piperazine Derivatives:

Unless otherwise started, substituted piperazine were mono-benzoylated using the following procedures:

Preparation of Benzoylpiperazines XXVIII and XXIX

To a stirred solution of substituted piperazine (1.0 g, 11.6 mmol) in dry THF (50 ml) under argon was added 2.5M n-BuLi in THF (10.23 ml, 25.5 mmol) at room temperature. After stirring for 1 hour at room temperature, benzoyl chloride (1.27 ml, 11.0 mmol) was added to the solution of dianion and the reaction mixture was stirred for an additional 10 minutes. The reaction mixture was quenched with MEOH, and the solvents evaporated. The residue was partitioned between EtOAc (50 ml) and sat. NaHCO

3

. The aqueous layer was saturated with NaCl and extracted with EtOAc (2×30 ml). The organic layer was dried over MgSO

4

and concentrated to afford the crude product benzoylpiperazine, which was generally of sufficient purity to be used directly without further purification. Chromatography on a silica gel column (EtOAc/MeOH/Et

3

N, 7:3:1) gave the purified product.

Preparation of Benzoylpiperazine XXXIII and XLIII

To a stirred solution of 2-isopropylpiperazine (1.0 g, 7.81 mmol) in dry THF (50 mL), maintained at room temperature under argon atmosphere, was added a solution of 2.5 M n-BuLi in THF (6.88 mL, 17.2 mmol). After stirring for 30 minutes at room temperature, benzoyl chloride (0.86 ml, 7.42 mmol) was added and the reaction mixture stirred for an additional 10 minutes. The reaction mixture was then quenched with MeOH, the solvents were evaporated in vacuo and the residue was purified by silica gel flash chromatography. Elution with a mixture of EtOAc and MeOH (1:1) afforded product XXXIII (0.62 g, 36% yield) and XLIII (0.3 g, 17% yield). Benzoyl piperazines XXXIII, XXXIV, XXXV, XXXVI, XXXVII were prepared using the same procedure as that outlined above.

Preparation of Benzoylpiperazines XXXI, XXXII, XXXVIII,

Commercially available benzoic acid (4.8 g, 40 mmol), pentafluorophenol (7.4 g, 40 mmol) and EDAC (7.6 g, 40 mmol) were combined in 60 ml of dry DMF. The mixture was stirred at room temperature for 2 hours. To this solution, 2-methylpiperazine (4.0 g, 40 mmol) in 30 ml of DMF was added slowly and the reaction mixture was stirred at room temperature for 12 hours.

Evaporation of DMF gave a residue which was diluted with 400 ml of EtOAc and washed with water (2×100 nm). The organic phase was dried over anhydrous MgSO4 and concentrated in vacuo to provide a crude product, which was purified by column chromatography with EtOAc/MeOH (100:1) and then EtOAc/MeOH (10:1) to give 4.8 g of product XX in 60% yield.

Preparation of Benzoylpiperazine XXX

To a stirred solution of 2-methylpiperazine (10.0 g, 0.1 mol) in dry CH

2

Cl

2

(500 ml) under argon was added a solution of 1.0 M Me

2

AlCl or Et

2

AlCl in hexanes (100 ml, 0.1 mmol) and methyl benzoate (12.4 ml, 0.1 mmol) at room temperature. The reaction mixture was then stirred for 2 days before 2N NaOH (200 ml) was added. Aqueous layer was extracted with EtOAc (3×100 ml). The combined organic layer was dried over MgSO

4

and concentration of solution provided 20.0 g of crude product (98%), with was pure enough for the further reactions.

Preparation of N-Benzoyl-cis-2,6-dimethylpiperazine XLVII

To a stirred solution of 2,6-di-methylpiperazine (0.82 g, 7.2 mmol) in dry THF (50 mL), maintained at room temperature under an argon atmosphere, was added a solution of 2.5 M n-BuLi in THF (6.3 mL, 15.8 mmol). After stirring for 30 minutes at room temperature, trimethylsilyl chloride (1.0 mL, 7.9 mmol) was added and the reaction mixture stirred for one hour before the addition of benzoyl chloride (0.80 mL, 6.9 mmol). After 10 minutes, the reaction mixture was quenched with MeOH and the solvents were evaporated in vacuo. The residue was purified by silica gel flash column chromatography eluting with a mixture of EtOAc and MeOH (1:1) to provide product XLVII (1.48 g, 99% yield). Benzoyl piperazines XL, XLI, XLII, XLIII, XLIV, XLV, and XLVI were synthesised using the same procedure as outlined above.

Preparation of Benzoylpiperazine XXXIX

To a stirred solution of 2-ethoxycarbonylpiperazine (4.6 g, 29.1 mmol) in dry methylene chloride (200 mL), was added benzoyl chloride (3.55 ml, 29.1 mmol) and triethylamine (2 ml) sequentially. After stirring for 8 hours at room temperature, a saturated NaHCO

3

solution was added and the aqueous phase was extracted with ethyl acetate (3×200 ml). The organic layers were combined, dried over MgSO

4

and concentrated to give a crude mixture, which included the desired product XXXIX. The crude was then used for the further reaction without purification.

Preparation of Benzoylpiperazine XLVIII

To a stirred solution of 2-methoxycarbonyltetrahydropyrazine (1.0 g, 7.0 mmol) in dry methylene chloride (50 ml), was added benzoyl chloride 0.76 ml, 6.7 mmol) and triethylamine (5 ml) sequentially. After stirring for 8 hours at room temperature, a saturated NaHCO

3

solution was added and the aqueous phase was extracted with ethyl acetate (3×20 ml). The organic layers were combined, dried over MgSO

4

and concentrated to give a crude mixture, which included the desired product XLVII. The crude was then used for the further reaction without purification.

Preparation of 3-Hydroxylmethyl-benzoylpiperazine XLIX

To a stirred solution of 3-ethoxycarbonyl-benzoylpiperazine XLIX (200 mg, 0.76 mmol) in THF (5 ml), was added lithium chloride (36 mg, 0.84 mmol), NaBH4 (32 mg, 0.84 mmol) and EtOH (5 ml) sequentially. After stirring for 8 hours at room temperature, a saturated NaHCO

3

solution was added and the aqueous phase was extracted with ethyl acetate (3×20 ml). The organic layers were combined, dried over MgSO

4

and concentrated to give a crude mixture, which was used for the further reaction without purification.

Characterization of Mono-Benzoylated Piperazine Derivatives

N-Benzoylpiperazine XXVIII:

1

H NMR (300 MHz, CD

3

OD) δ 7.37 (m, 5H), 3.73 (br s, 2H), 3.42 (br s, 2H), 2,85 (br s, 4H);

13

C NMR (75 MHz, CD

3

OD) δ 170.9, 135.0, 129.6, 128.2, 126.5, 44.5; HRMS m/z: (M+H)

+

calcd for C

11

H

15

N

2

O 191.1184, found 191.1181.

N-(Benzoyl)-trans-2,5-Dimethylpiperazine XXIX.

1

H NMR (300 MHz, CD

3

OD) δ 7.50-7.28 (m, 5H), 4.38 (br s, 1H), 3.70 (br s, 1H), 3.40-3.20 (m, 3H), 2.57 (dd, 1H, J=12.96, 1.98 Hz), 1,35 (d, 3H, J=6.87 Hz), 1.22 (d, 3H, J=6.78 Hz);

13

C NMR (75 MHz, CD

3

OD) δ 171.9, 135.9, 129.3, 128.3, 126.0, 47.6, 46.7, 43.8, 42.3, 14.7, 14.3; HRMS m/z: (M+H)

+

calcd for C

13

H

19

N

2

O 219.1497, found 219.1499.

N-(Benzoyl)-3-methylpiperazine XXX.

1

H NMR (300 MHz, CD

3

OD) δ 7.45 (m, 5H), 4.50 (d, 1H, J=10.8 Hz), 3.60 (b, 1H), 3.33-2.60 (m, 5H), 1.16-0.98 (m, 3H);

13

C NMR (75 MHz, CD

3

OD) δ 170.9, 135.3, 129.6, 128.3, 126.5, 54.0, 50.6, 50.1, 45.0, 44.4, 41.7, 17.50; HRMS m/z: (M+H)

+

calcd for C

12

H

17

N

2

O 205.1341, found 205.1336.

N-(Benzoyl)-3-ethylpiperazine XXXI.

1

H NMR (300 MHz, CD

3

OD) δ 7.47 (m, 5H), 4.55 (b, 1H), 3.64 (b, 1H), 3.36-2.59 (m, 5H), 1.51-0.82 (m, 5H);

13

C NMR (75 MHz, CD

3

OD) δ 171.5, 135.8, 130.1, 128.8, 126.9, 57.2, 56.7, 52.9, 47.1, 45.5, 42.5, 26.4, 26.0, 9.3; HRMS m/z: (M+H)

+

calcd for C

13

H

19

N

2

O 219.1497, found 219.1495.

N-(Benzoyl)-3-propylpiperazine XXXII.

1

H NMR (300 MHz, CD

3

OD) δ 7.45 (m, 5H), 4.53 (t, 1H, J=13.44 Hz), 3.64 (b, 1H), 3.17-2.64 (m, 5H), 1.46-0.86 (m, 7H);

13

C NMR (75 MHz, CD

3

OD) δ 171.4, 135.9, 130.1, 128.8, 126.9, 55.4, 54.9, 53.2, 45.6, 45.0, 42.6, 35.8, 35.3, 18.8, 13.4; HRMS m/z: (M+H)

+

calcd for C

14

H

21

N

2

O 233.1654, found 233.1652.

N-(Benzoyl)-3-iso-propylpiperazine XXXIII.

1

H NMR (300 MHz, CD

3

OD) δ 7.45 (m, 5H), 4.30 (m, 1H), 3.64 (m, 1H), 3.10-2.40 (m, 5H), 1.70-0.75 (m, 7H);

13

C NMR (75 MHz, CD

3

OD) δ 171.5, 135.9, 130.5, 129.3, 126.9, 61.7, 61.1, 51.2, 45.9, 45.4, 42.5, 31.2, 30.7, 18.3; HRMS m/z: (M+H)

+

calcd for C

14

H

21

N

2

O 233.1654, found 233.1654.

N-(Benzoyl)-3-pentylpiperazine XXXIV.

1

H NMR (300 MHz, CD

3

OD) δ 7.47 (m, 5H), 4.50 (t, 1H, J=17.85 Hz), 3.62 (b, 1H), 3.17-2.64 (m, 5H), 1.46-0.87 (m, 11H);

13

C NMR (75 MHz, CD

3

OD) δ 171.4, 135.9, 130.1, 129.3, 126.8, 55.6, 55.2, 53.1, 45.6, 45.0, 42.5, 33.6, 33.0, 32.0, 28.9, 25.9, 25.3, 22.6, 13.4; HRMS m/z: (M+H)

+

calcd for C

16

H

25

N

2

O 261.1967, found 261.1969.

N-(Benzoyl)-3-iso-butylpiperazine XXXV. MS m/z: (M+H)

+

calcd for C

15

H

23

N

2

O: 247.18, found 247.22. HPLC retention time: 1.04 minutes (Method C).

N-(Benzoyl)-3-tert-butylpiperazine XXXVI.

1

H NMR (300 MHz, CD

3

OD) δ 7.45 (m, 5H), 4.70 (m, 1H), 3.66 (m, 1H), 3.17-2.43 (m, 5H), 1.17-0.84 (m, 9H);

13

C NMR (75 MHz, CD

3

OD) δ 171.6, 135.9, 131.0, 129.4, 126.9, 65.3, 64.6, 49.6, 46.5, 45.9, 43.7, 42.3, 32.7.25.7; HRMS m/z: (M+H)

+

calcd for C

15

H

23

N

2

O 247.1810, found 247.1815.

N-(Benzoyl)-cis-3,5-di-methylpiperazine XXXVII.

1

H NMR (300 MHz, CD

3

OD) δ 7.43 (m, 5H), 4.55 (d, 1H,J=12.0 Hz), 3.55 (d, 1H, J=9.60 Hz), 2.74-2.38 (m, 5H), 1.13-0.94 (m, 6H);

13

C NMR (75 MHz, CD

3

OD) δ 170.5, 135.5, 129.6, 128.3, 126.6, 53.4, 50.9, 50.2, 17.7, 17.3; HRMS m/z: (M+H)

+

calcd for C

13

H

19

N

2

O 219.1497, found 219.1492.

N-(Benzoyl)-3-trifluoromethylpiperazine XXXVIII. MS m/z: (M+H)

+

calcd for C

12

H

14

F

3

N

2

O: 259.11, found 259.05. HPLC retention time: 0.65 minutes (Method A).

N-(Benzoyl)-3-ethoxycarbonylpiperazine XXXIX. MS m/z: (M+H)

+

calcd for C

14

H

19

N

2

O

3

: 263.14, found 263.20. HPLC retention time: 0.80 minutes (Method C).

N-(Benzoyl)-2-methylpiperazine XL.

1

H NMR (300 MHz, CD

3

OD) δ 7.47 (m, 5H), 3.30-2.70 (m, 7H), 1.36 (d, 3H, J=6.90 Hz);

13

C NMR (75 MHz, CD

3

OD) δ 171.0, 135.4, 129.7, 128.5, 126.3, 48.5, 44.3, 14.5; HRMS m/z: (M+H)

+

calcd for C

12

H

17

N

2

O 205.1341, found 205.1341.

N-(Benzoyl)-2-ethylpiperazine XLI.

1

H NMR (300 MHz, CD

3

OD) δ 7.49 (m, 5H), 3.34-2.80 (m, 7H), 2.10-1.70 (m, 2H), 0.85 (b, 3H);

13

C NMR (75 MHz, CD

3

OD) δ 171.5, 135.1, 129.8, 128.5, 126.5, 48.5, 46.0, 43.9, 21.8, 9.6; HRMS m/z: (M+H)

+

calcd for C

13

H

19

N

2

O 219.1497, found 219.1501.

N-(Benzoyl)-2-propylpiperazine XLII.

1

H NMR (300 MHz, CD

3

OD) δ 7.50 (m, 5H), 3.60-2.80 (m, 7H), 2.10-0.70 (m, 7H); 13C NMR (75 MHz, CD

3

OD) δ 172.5, 135.0, 129.9, 128.6, 126.7, 48.7, 46.2, 43.8, 30.9, 18.9, 13.1; HRMS m/z: (M+H)

+

calcd for C

14

H

21

N

2

O 233.1654, found 233.1650.

N-(Benzoyl)-2-iso-propylpiperazine XLIII.

1

NMR (300 MHz, CD

3

OD) δ 7.50 (b, 5H), 4.40 (m, 1H), 3.60-2.50 (m, 6H), 1.10-0.70 (m, 7H);

13

C NMR (75 MHz, CD

3

OD) δ 171.1, 135.0, 130.0, 128.7, 127.0, 60.6, 54.1, 43.9, 42.3, 25.4, 19.3, 18.4; HRMS m/z: (M+H)

+

calcd for C

14

H

21

N

2

O 233.1654, found 233.1653.

N-(Benzoyl)-2-pentylpiperazine XLIV.

1

H NMR (300 MHz, CD

3

OD) δ 7.47 (b, 5H), 3.40-2.80 (m, 7H), 2.10-0.70 (m, 11H); 13C NMR (75 MHz, CD

3

OD) δ 71.2, 135.0, 129.9, 128.6, 126.7, 48.7, 46.2, 43.8, 31.0, 28.8, 25.3, 22.2, 13.4; HRMS m/z: (M+H)

+

calcd for C

16

H

25

N

2

O 261.1967, found 261.1970.

N-(Benzoyl)-2-iso-butylpiperazine XLV. MS m/z: (M+H)

+

calcd for C

15

H

23

N

2

O: 247.18, found 247.23. HPLC retention time: 1.06 minutes (Method C).

N-(Benzoyl)-2-tert-butylpiperazine XLVI.

1

H NMR (300 MHz, CD

3

OD) δ 7.45 (m, 5H), 4.53 (t, 1H, J=5.70 Hz), 3.60-2.60 (m, 6H), 1.14 (s, 9H);

13

C NMR (75 MHz, CD

3

OD) δ 173.5, 136.7, 129.9, 128.9, 126.6, 55.9, 44.8, 44.5, 42.7, 36.5, 27.8; HRMS m/z: (M+H)

+

calcd for C

15

H

23

N

2

O 247.1810, found 247.1808.

N-(Benzoyl)-cis-2,6-di-methylpiperazine XLVII.

1

H NMR (300 MHz, CD

3

OD) δ 7.45 (m, 5H), 4.18 (b, 2H), 2.85 (m, 4H), 1.33 (d, 6H, J=6.90 Hz);

13

C NMR (75 MHz, CD

3

OD) δ 172.0, 136.7, 128.9, 128.3, 125.8, 49.1, 47.1, 19.2; HRMS m/z: (M+H)

+

calcd for C

13

H

19

N

2

O 219.1497, found 219.1491.

N-(Benzoyl)-3-methoxycarbonyltetrahydropyrazine XLVIII. MS m/z: (M+H)

+

calcd for C

13

H

15

N

2

O

3

: 247.11, found 247.13. HPLC retention time: 1.00 minutes (Method C).

3-Hydroxylmethyl-benzoylpiperazine XLIX. MS m/z: (M+H)

+

calcd for C

12

H

17

N

2

O

2

: 221.13, found 221.17. HPLC retention time: 0.32 minutes (Method C).

C. Coupling of Mono-benzoyl Piperazines with Glyoxyl Chlorides

To a solution of indole glyoxoyl chloride V (1 eq) in dry CH

2

Cl

2

was added substituted benzoylpiperazine (1 eq) at room temperature. The mixture was then cooled down to 0° C., followed by dropwise addition of diisopropylamine (1.3 eq). After 5 min., the reaction mixture was warmed to room temperature and was shaken for 3 hr. The resulting crude products XL were purified by preparative HPLC and characterized as shown in Table 6.

Preparation of N-(Benzoyl)-3-hydroxylmethyl-N′-[(7-methoxycarbonyl-indol-3-yl)-oxoacetyl]-piperazine Example 98

To a stirred solution of 3-hydroxylmethyl-benzoylpiperazine XLIX (8.0 mg, 0.036 mmol) in acetonitrile (5 ml) was added BSTFA (8.1 mg, 0.036 mmol). After stirring for 30 minutes at room temperature, (7-methoxycarbonyl-indol-3-yl)-oxoacetyl chloride (8.1 mg, 0.036 mmol) and pyridine (0.5 ml) were added. The reaction was stirred for another 2 hours at room temperature. Concentration under vaccum provided a residue, which was then purified by Shimazu HPLC purification system to give 2 mg of N-(benzoyl)-3-hydroxylmethyl-N′-[(7-methoxycarbonyl-indol-3-yl)-oxoacetyl]-piperazine (Example 98).

D. Hydrolysis of Ester Group to Acid Group:

Preparation of N-(Benzoyl)-3-hydroxycarbonyl-N′-[(4-fluoro-indol-3-yl)-oxoacetyl]-piperazine (Example 101)

To a stirred solution of N-(benzoyl)-3-ethoxycarbonyl-N′-[(4-fluoro-indol-3-yl)-oxoacetyl]-piperazine (100 mg, 0.02 mmol) in methanol (1 ml) and water (1 ml), was added potassium carbonate (9 mg, 0.06 mmol). After stirring for 8 hours at room temperature, the product was concentrated in vacuo to give a residue which was purified by preparative HPLC to yield 2 mg of N-(benzoyl)-3-hydroxycarbonyl-N′-[(4-fluoro-indol-3-yl)-oxoacetyl]-piperazine (Example 101).

Preparation of N-(Benzoyl)-3-(R)-methyl-N′-[(7-hydroxycarbonyl-indol-3-yl)-oxoacetyl]-piperazine (Example 137)

To a stirred solution of N-(benzoyl)-3-(R)-methyl-N′-[(7-methoxycarbonyl-indol-3-yl)-oxoacetyl]-piperazine (20 mg, 0.05 mmol) in 0.5N sodium methoxide in methanol (5 ml), was added 0.5 ml of water. After stirring for 8 hours at room temperature, 10% HCl was added to the reaction mixture to pH=6. N-(benzoyl)-3-(R)-methyl-N ′-[(7-hydroxycarbonyl-indol-3-yl)-oxoacetyl]-piperazine (Example 137) precipated out from the solution, which was collected via filtration.

TABLE 6

HPLC

Exam-

retention

ple

time

MS Data

#

R1

W

(min)

(M+H)

+

91

4-Fluoro

1.27

A

394

92

4-Fluoro

1.32

A

408

93

4-Fluoro

0.92

B

394

94

4-Fluoro

1.32

A

394

95

4-Fluoro

1.32

A

394

96

4-Fluoro

1.29

A

408

97

4-Fluoro

1.29

A

408

98

4-COOMe

1.44

A

450

99

4-Fluoro

1.23

A

432

100

7-COOMe

1.60

A

492

101

4-Fluoro

1.25

A

424

102

7-COOMe

1.41

B

434

103

7-COOMe

1.39

A

434

104

7-COOMe

1.57

A

434

105

7-OMe

1.55

406

106

4,7-difluoro

1.33

412

107

4,5,6,7- tetrafluoro

1.57

448

108

4,5,6,7- tetrafluoro

1.58

448

109

7-Nitro

1.48

421

110

7-Ethyl

1.58

404

111

7-OMe

1.39

406

112

7-Nitro

1.49

421

113

6-Chloro

1.55

411

114

5,6-dichloro

1.71

446

115

4-Chloro

1.45

410

116

4-Chloro

1.45

410

117

5,6-dichloro

1.72

446

118

5-Fluoro

1.43

394

119

7-Ethyl

1.55

1.67

120

4-Bromo

1.48

456

121

7-COOMe

1.62

A

476

122

4-Br

1.48

456

123

5-Fluoro

1.42

394

124

6-Chloro

1.57

410

125

7-COOMe

1.61

A

448

126

7-COOMe

1.69

A

462

127

7-COOMe

1.67

A

462

128

7-COOMe

1.69

A

462

129

7-COOMe

1.76

A

476

130

7-COOMe

1.76

A

476

131

7-COOMe

1.71

A

476

132

7-COOMe

1.84

A

490

133

7-COOMe

1.85

A

490

134

7-COOMe

1.76

A

476

135

4-OMe

1.24

406

136

4-OMe

1.24

406

137

7-COOH

1.43

A

420

138

4-Fluoro-7- methyl

1.37

408

139

7-COOMe

1.62

A

448

140

7-Fluoro

1.74

394

141

7-Fluoro

1.74

394

142

7-COOMe

1.65

A

488

143

4-Fluoro

1.42

A

488

144

4-fluoro-7- bromo

1.12

N.D

145

4-Fluoro-7- COOMe

1.51

452

146

4-Fluoro-7- COOMe

1.39

438

147

4-Fluoro-7- OMe

1.31

424

Note in Table 6, and other tables herein, in the HPLC column, numbers with superscript “A”, “B,” or “C” refer to the HPLC method used (i.e. Methods A, B or C, respectively).

Preparation of Examples 148 -194 in Table 7

Step A.

To substituted indole-3-glyoxylyl chloride V (1 eq) in CH

2

Cl

2

at room temperature was added tert-butyl 1-piperazinecarboxylate (1 eq) and diisopropylethylamine (1.5 eq). The solution was stirred for 2 hr at room temperature after which time LC/MS analysis indicated the completion of the reaction. The solvent was removed in vacuo and the resulting residue was diluted with ethyl acetate (250 ml) and diethylether (250 ml). The organic solution was then washed with water (100 ml×3) and brine (50 ml), dried over MgSO

4

, filtered and concentrated. To the light-yellow solid was then added 30 ml of 20% trifluoroacetic acid in CH

2

Cl

2

. The solution was concentrated and dried in vacuo to give the desired product VII. LC/MS analysis indicated this product was 100% pure and it was used for the next reaction without further purification.

Step B.

To piperazine indole-3-glyoxylamide (1 eq) was added resin-bound 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (P-EDC) (7 eq) and carboxylic acid (RCOOH) (2 eq) in dichloroethane (DCE) or DMF(dimethylfornamide) in cases where the carboxylic acids are not soluble in DCE. The reaction was shaken for 12 hr at room temperature. The product XLI was filtered and concentrated. Products with purity less than 70% were diluted in methanol and purified using a Shimadzu automated preparative HPLC System.

Preparation of Example 195 in Table 7

Step A

To a solution of tert-butyl-1-piperazine carboxylate (601 mg, 3.23 mmol) and 4-acetic acid imidazole (330 mg, 2.94 mmol) in dichloromethane (30 ml), were added DMAP (394 mg, 3.22 mmol) and EDC (616 mg, 3.22 mmol). The reaction mixture was stirred at room temperature for 21.5 hours. Removal of solvent in vacuo afforded a white solid, which was subjected to flash chromatography using a gradient elution (100% EtOAc, to 2% to 5% MeOH/EtOAc, to 1/5/95 NH

3

(sat. aq.)/MeOH/EtOAc) to give XLII as a white solid.

Step B

To compound XXLII (130 mg, 0.464 mmol) was added a solution of HCl in dioxane (4 M, 5 ml), and the mixture stirred at room temperature for 3 hours. Removal of the excess reagent in vacuo afforded the hydrochloride salt XLIII as a white solid (100% conversion).

Step C

4-Fluoroindole glyoxyl chloride was coupled with XLIII as described previously.

TABLE 7

HFLC

Retention

Time

MS Data

Entry

R

Ar

(min)

(M+H)

+

148

4-Fluoro

3-Thiophenyl

1.74

386

149

4-Fluoro

1,2,3-Thiadiazolyl

1.52

388

150

4-Fluoro

2-(4-Methoxy)-

1.78

416

thiophenyl

151

4-Fluoro

2-(5-Methylthio)-

2.28

432

thiophenyl

152

4-Fluoro

2-(3-Bromo)-

2.22

465

thiophenyl

153

4-Fluoro

2-(5-Bromo)-

2.29

465

thiophenyl

154

4-Fluoro

2-Pyrazinyl

1.76

382

155

4-Fluoro

2-(5-Methyl)-

2.24

400

thiophenyl

156

4-Fluoro

2-(5-Chloro)-

2.08

421

thiophenyl

157

4-Fluoro

2-Indolyl

2.07

419

158

4-Fluoro

4-(2-Methyl)-

2.00

401

thiazolyl

159

4-Fluoro

4-Thiazolyl

1.05

387

160

4-Fluoro

4-Pyridyl

0.84

381

161

4-Fluoro

3-(6-Methyl)-pyridyl

0.87

395

162

4-Fluoro

3-Pyridyl

0.93

381

163

4-Fluoro

5-Isoxazolyl

1.08

371

164

4-Fluoro

2-Furanyl

1.17

370

165

4-Fluoro

3-Pyrazolyl

1.03

370

166

4-Fluoro

2-Pyridyl

1.08

381

167

4-Fluoro

3-Furanyl

1.14

370

168

4-Fluoro

2-Thiophenyl

1.24

386

169

4-Fluoro

2-Benzofuranyl

1.48

420

170

4-Fluoro

2-(5-Bromo)-furanyl

1.37

449

171

4-Fluoro

2-(3-Methyl)-furanyl

1.30

384

172

4-Fluoro

2-(3-Chloro)-

1.34

420

thiophenyl

173

4-Fluoro

3-(5-Chloro-4-

1.45

451

methoxy)-

thiophenyl

174

4-Fluoro

2-(5-Chloro)-furanyl

1.32

404

175

4-Chloro

3-Thiophenyl

2.02

403

176

4-Chloro

2-[5-(Pyrid-2-yl)]-

2.07

480

thiophenyl

177

4-Chloro

2-Thieno[3,2-B]-

2.33

459

thiophenyl

178

4-Chloro

2-(5-Methylthio)-

2.33

449

thiophenyl

179

4-Chloro

2-(5-Bromo)-

2.34

481

thiophenyl

180

4-Chloro

2-Pyrazinyl

1.91

398

181

4-Chloro

2-Pyridyl

1.92

397

182

4-Chloro

2-Benzothiophenyl

2.36

453

183

4-Chloro

2-(5-Chloro)-

2.33

437

thiophenyl

184

4-Chloro

2-(3-Chloro)-

2.27

437

thiophenyl

185

4-Chloro

2-Indolyl

2.33

436

186

4-Chloro

4-(2-Methyl)-

2.22

418

thiazolyl

187

4-Chloro

4-Thiazolyl

1.20

404

188

4,7-Difluoro

2-(5-Chloro)-furanyl

1.39

422

189

4,7-Difluoro

2-(5-Bromo)-furanyl

1.46

467

190

4,7-Difluoro

2-furanyl

1.28

388

191

4,7-Difluoro

2-Pyridyl

1.17

399

192

4,7-Difluoro

2-(3,4-Dichloro)-

1.47

457

furanyl

193

4,7-Difluoro

2-(5-

1.54

456

Trifluoromethyl)-

furanyl

194

4,7-Difluoro

2-(4,5-Dimethyl)-

1.49

416

furanyl

195

4-Fluoro

2-Imidazolyl

0.81

370

Synthesis of Examples 195-215 in Table 8

Step A

To pentafluorophenol (1.84 g, 10 mmol) in DMF (15 mL) was added picolinic acid (1.23 g, 10 mmol) and EDC (1.91 g, 10 mmol) at room temperature for 4 h. The crude product XLIV was diluted with CH

2

Cl

2

and was washed with water, 0.1 M HCl and brine. The organic phase was dried over MgSO4, filtered and concentrated. The crude material was used without further purification.

Step B

To a solution of (R)-methyl pierazine (1.0 g, 10 mmol) in DMF (20 mL) at room temperature was slowly added a solution of picolinic acid pentafluorophenylester XLIV in DMF (20 mL). The reaction mixture was stirred at room temperature for 16 h. The product was diluted with CH2Cl2 and was washed with water and brine, dried over MgSO4, filtered and concentrated. The product XLV was then purified by flash chromatography (100% EtOAc—50% MeOH/EtOAc).

Piperazine XLVI was prepared using similar methodology to that outlined in Step A and Step B above.

Step C

To the mixture of indole glyoxylchloride V (1 eq) and 3-(R)-methyl-1-piperazinecarboxylate XLV or XLVI (1 eq) in THF was added diisopropylethylamine (1.5 eq) dropwise at 0° C. The solution was stirred for additional 2 hr at room temperature and the resulting crude compounds were purified by preparative HPLC.

TABLE 8

HPLC

Retention

Time

MS Data

Entry

R

Ar

(min)

(M+H)

+

196

4,7-

2-Pyridyl

1.23

413

difluoro

197

4-Fluoro-7-

2-Pyridyl

1.17

409

methyl

198

4,7-

2-(5-Bromo)-

1.52

481

Difluoro

furanyl

199

4,7-

2-(5-Bromo)-

1.45

506

Dimethoxy

furanyl

200

7-COOMe

2-(5-Bromo)-

1.70

504

furanyl

201

4,7-

2-Pyridyl

1.23

413

Difluoro

202

4-Fluoro

2-Pyridyl

1.07

395

203

4-Chloro

2-Pyridyl

1.22

411

204

4-Bromo

2-Pyridyl

1.25

457

205

5-Fluoro

2-Pyridyl

1.21

395

206

6-Chloro

2-Pyridyl

1.43

411

207

7-Fluoro

2-Pyridyl

1.29

395

208

7-Methoxy

2-Pyridyl

1.26

407

209

7-Methyl

2-Pyridyl

1.31

391

210

7-Ethyl

2-Pyridyl

1.46

403*

211

4-methoxy-

2-Pyridyl

1.22

441

7-chloro

212

7-cyano

2-Pyridyl

1.24

402

213

4-Methoxy

2-Pyridyl

1.09

407

214

4-

2-Pyridyl

1.28

487

Methoxy-

7-Bromo

215

4-Fluoro-7-

2-Pyridyl

1.16

425

Methoxy

*(M-H) measured in negative ionization mode

Additional Analytical Data for Selected Compounds

1-(4-Methylbenzoyl)-4-[(1H-indol-3-yl)oxoacetyl]piperazine (Example 15)

MS (ESI): 376 (M+H)

+

; IR (KBr): 3150, 3104, 2922, 2868, 1780, 1629, 1519, 1433, 1272, 1158, 1006, 829, 775, 753, 645 cm

−1

;

1

H NMR (CDCl

3

) δ 2.40 (s, 3H), 3.60-3.79 (m, 8H), 7.23-7.45 (m, 7H), 7.99 (d, J=3.1 Hz, 1H), 8.34 (m, 1H), 9.10 (s, 1H).

1-(Benzoyl)-4-[(1H-4-fluoroindol-3-yl)oxoacetyl]piperazine (Example 19)

MS (ESI): 380 (M+H)

+

HRMS calcd for C

21

H

18

FN

3

O

3

[(M+H)]

+

, 380.14105; found, 380.1412.

1

H-NMR (DMF-d7) δ 12.71(s, 1H), 8.02 (s, 1H), 7.46-7.56 (m, 6H), 7.31 (ddd, J=4.71, 7.99 Hz, 1H), 7.03 (dd, J=7.84, 10.98 Hz, 1H), 3.77 (br. s, 4H), 3.57 (br. s, 4H).

1-(Benzoyl)-4-[(1H-4-chloroindol-3-yl)oxoacetyl]piperazine (Example 20)

MS (ESI): 396 (M+H)

+

HRMS calcd for C

21

H

18

ClN

3

O

3

[(M+H)]

+

, 396.11150; found, 396.1105. 1H NMR (DMF-d7) δ 12.74 (br. s, 1H), 8.03 (s,1H), 7.57-7.65 (m, 1H), 7.50 (s, 5H), 7.28-7.38 (m, 1H), 3.42-3.83 (m, 8H).

1-(Benzoyl)-4-[(1H-6-fluoroindol-3-yl)oxoacetyl]piperazine (Example 28)

MS (ESI): 380 (M+H)

+

HRMS calcd for C

21

H

18

FN

3

O

3

[(M+H)]

+

, 380.14105; found, 380.1414. 1H NMR (DMF-d7) δ 12.09 (s, 1H), 7.81 (dd, J=5.64, 8.46 Hz, 1H), 7.62(s, 1H), 7.08 (s, 5H), 7.01 (dd, J=2.28, 9.61 Hz, 1H), 2.86-3.52 (m, 8H). Anal. Calcd for C

21

H

18

FN

3

O

3

: C, 66.48; H, 4.78, N, 11.08. Found: C, 66.09, H, 4.78, N, 10.94.

1-(Benzoyl)-4[(1H-4,6-difluoroindol-3-yl)oxoacetyl]piperazine (Example 42)

1

H NMR (DMSO-d

6

) δ 3.40 (br s, 4H), 3.65 (br s, 4H), 7.06 (t, 1H), 7.20 (d, J=8.49 Hz, 1H), 8.27 (s, 1H), 12.65 (br s, 1 H). Anal. Calcd for C

21

H

17

F

2

N

3

.O

3

0.322 H

2

O: C, 62.57; H, 4.41; N, 10.42; Found: C, 62.56; H, 4.46; N, 10.11.

1-(benzoyl)-4-[(1H-5-fluoro-7-bromoindol-3-yl)oxoacetyl]piperazine (Example 48)

1

H NMR (DMSO-d

6

) δ 3.40 (br s, 4H), 3.67 (br s, 4H), 7.43 (br s, 5H), 7.54 (dd, J=2.25, 8.97 Hz, 1H), 7.83 (d, J=8.4 Hz, 1H), 8.29 (s, 1H), 12.79 (br s, 1 H). Anal. Calcd for C

21

H

17

N

3

BrF

3

.1.2 H

2

O: C, 52.56; H, 4.07; N, 8.76. Found: C, 52.33; H, 3.69; N, 8.50.

1-(benzoyl)-4-[(1H-4-fluoro-7-trifluoroethoxyindol-3-yl)oxoacetyl]piperazine (Example 51)

1

H NMR (DMSO-d

6

) δ 3.61 (br m, 4H), 3.80 (br m, 4H), 4.52 (m, 2H), 6.68 (m, 1H), 6.91 (m, 1H), 7.45 (s, 5H), 8.07 (d, J=2.91 Hz, 1H), 9.37 (s 1H). Anal. Calcd for C

23

H

19

F

4

N

3

O

4

.0.59 H

2

O, 0.47 ethyl acetate C, 56.44; H, 4.56; N,=7.94; Found: C, 56.44; H, 4.16; N,=8.19.

1-(Benzoyl)-4-[(1H-4-bromo-7-fluoroindol-3-yl)oxoacetyl]piperazine (Example 55)

1

H NMR (CDCl

3

) δ 3.6-3.9 (br m, 8H), 6.92 (t, 1H), 7.42 (br s, 6H), 8.09 (s, 1H), 9.5 (br s, 1 H). Anal. Calcd for C

21

H

17

BrFN

3

O

3

.0.25 H

2

O, 0.21 ethyl acetate: C, 54.5; H, 4.02; N, 16.6; Found: C, 54.50; H, 4.09; N,

8.44. Example

90

1

H NMR: (DMSO-d

6

) δ 3.66 (br. s, 4H), 7.27 (t, J=8.31 Hz, 1H), 7.43 (br. s, 7H), 8.01 (m, 1H), 8.14 (s, 1H), 10.14 (s, 1H), 12.42 (br. s, 1H) MS: (M+H)

+

480.00, (M−H) 406.02; IR: 1636,1592 cm

−1

.

N-(Benzoyl)-(R)-3-methyl-N′-[(4-fluoro-indol-3-yl)-oxoacetyl]-piperazine (Example 93)

1

H NMR (300 MHz, CD

3

OD) δ 8.20 (s, 0.5H), 8.15 (s, 0.5H), 7.48-6.90 (m, 8H), 5.00-3.00 (m, 7H), 1.30 (b, 3H). MS m/z: (M+H)

+

calcd for C

22

H

21

FN

3

O

3

: 394.16; found 394.23. HPLC retention time: 0.92 minutes (Method B).

N-(Benzoyl)-(S)-3-methyl-N′-[(fluoro-indol-3-yl)-oxoacetyl]-piperazine (Example 94)

1

H NMR (300 MHz, CD

3

OD) δ 8.20 (s, 0.5H), 8.13 (s, 0.5H), 7.48-6.90 (m, 8H), 5.00-3.00 (m, 7H), 1.30 (b, 3H). MS m/z: (M+H)

+

calcd for C

22

H

21

FN

3

O

3

: 394.16; found 394.25. HPLC retention time: 1.32 minutes (Method A).

N-(Benzoyl)-2-methyl-N′-[(4-fluoro-indol-3-yl)-oxoacetyl]-piperazine (Example 95)

1

H NMR (300 MHz, CD

3

OD) δ 8.20 (s, 0.5H), 8.13 (s, 0.5H), 7.48-6.90 (m, 8H), 5.00-3.00 (m, 7H), 1.37 (d, J=6.78 Hz, 1.5H), 1.27 (d, J=6.84 Hz, 1.5H). MS m/z: (M+H)

+

calcd for C

22

H

21

FN

3

O

3

: 394.16; found 394.23. HPLC retention time: 1.32 minutes (Method A).

N-(Benzoyl)-3-hydroxylmethyl-N′-[(4-fluoro-indol-3-yl)-oxoacetyl]-piperazine (Example 99)

1

H NMR (300 MHz, CD

3

OD) δ 7.50 (b, 5H), 7.39-6.72 (m, 4H), 5.00-2.80 (m, 9H). MS m/z: (M+Na)

+

calcd for C

22

H

20

FN

3

NaO

4

: 432.13; found 432.19. HPLC retention time: 1.23 minutes (Method A).

N-(benzoyl)-(R)-3-methyl-N′-[(7-methoxycarbonyl-indol-3-yl)-oxoacetyl]-piperazine (Example 102)

1H NMR (300 MHz, CD

3

OD) 8.50 (d, J=6.48 Hz, 1H), 8.15 (s, 0.5H), 8.10 (s, 0.5H), 8.00 (d, J=7.38 Hz, 1H), 7.42 (m, 6H), 5.00-3.00 (m, 7H), 4.02 (s, 3H), 1,34 (b, 3H);

13

C NMR (75 MHz, CD

3

OD); δ 186.2, 166.9, 137.9, 135.3, 130.3, 128.8, 127.2, 126.9, 126.4, 122.7, 114.5, 114.0, 51.6, 50.7, 45.6, 15.4, 14.2. MS m/z: (M+H)

+

calcd for C

24

H

24

N

3

O

5

: 434.17; found 434.24. HPLC retention time: 1.41 minutes (Method B).

N-(Benzoyl)-3-hydroxylmethyl-N′-[(7-methoxycarbonyl-indol-3-yl)-oxoacetyl]-piperazine (Example 98)

1

H NMR (300 MHz, CD

3

OD) δ 8.54 (b, 1H), 8.24 (s, 0.5H), 8.16 (s, 0.5H), 8.00 (m, 1H), 7.47 (m, 6H), 5.00-3.00 (m, 9H), 4.02 (s, 3H). MS m/z: (M+H)

+

calcd for C

24

H

24

N

3

O

6

: 450.17; found 450.24. HPLC retention time: 1.44 minutes (Method A).

N-(Benzoyl)-2-methoxycarbonyl-N′-[(7-methoxycarbonyl-indol-3-yl)-oxoacetyl]-tetrahydropyrazine (Example 121)

1

H NMR (500 MHz, CD

3

OD) δ 8.50 (m, 1H), 8.21 (s, 1H), 7.93 (m, 1H), 7.44 (m, 7H), 4.00 (s, 6H), 4.00-3.30 (m, 4H);

13

C NMR (125 MHz, CD

3

OD) δ 184.7, 167.9, 166.1, 165.3, 165.1, 164.9, 140.2, 137.2, 132.5, 129.6, 128.3, 128.2, 127.6, 125.1, 123.9, 116.0, 115.6, 115.0, 52.8, 52.6, 47.0, 43.8. MS m/z: (M+H)

+

calcd for C

25

H

22

N

3

O

7

: 476.15; found 476.21. HPLC retention time: 1.62 minutes (Method A).

N-(Benzoyl)-2-propyl-N′-[(7-methoxycarbonyl-indol-3-yl)-oxoacetyl]-piperazine (Example 126)

1

H NMR (300 MHz, CD

3

OD) δ 8.50 (d, J=6.93 Hz, 1H), 8.17 (s, 0.5H), 8.08 (s, 0.5H), 7.98 (d, J=6.00 Hz, 1H), 7.45 (m, 6H), 5.00-2.90 (m, 7H), 4.02 (s, 3H), 1.70-0.60 (m, 7H);

13

C NMR (75 MHz, CD

3

OD) 5186.0, 167.6, 166.9, 138.0, 136.1, 135.8, 130.2, 128.9, 127.2, 126.9, 126.4, 122.7, 114.5, 114.1, 51.7, 46.2, 44.0, 41.3, 31.5, 19.2, 13.2, 12.9. MS m/z: (M+H)

+

calcd for C

26

H

28

N

3

O

5

: 462.20; found 462.30. HPLC retention time: 1.69 minutes (Method A).

N-(Benzoyl)-(R)-3-methyl-N′-[(7-hydroxycarbonyl-indol-3-yl)-oxoacetyl]-piperazine (Example 137)

1

H NMR (300 MHz, CD

3

OD) δ 8.46 (b, 1H), 8.14 (s, 0.5H), 8.09 (s, 0.5H), 8.00 (d, J=7.17 Hz, 1H), 7.43 (m, 6H), 5.00-2.90 (m, 7H), 1.32 (b, 3H);

13

C NMR (75 MHz, CD

3

OD) δ 186.3, 168.2, 167.1, 137.8, 135.3, 130.3, 128.8, 127.1, 126.9, 126.7, 122.7, 115.4, 113.9, 50.7, 45.6, 15.4, 14.2. MS m/z: (M+H)

+

calcd for C

27

H

30

N

3

O

5

: 420.16; found 420.16. HPLC retention time: 1.43 minutes (Method A).

N-(Benzoyl)-3-trifluoromethyl-N′-[(7-methoxycarbonyl-indol-3-yl)-oxoacetyl]-piperazine (Example 142)

1

H NMR (300 MHz, CD

3

OD) δ 8.52 (b, 1H), 8.03 (s, 1H), 8.02 (d, J=7.56 Hz, 1H), 7.45 (m, 6H), 5.00-3.00 (m, 7H), 4.03 (s, 3H). MS m/z: (M+H)

+

calcd for C

24

H

21

F

3

N

3

O

5

: 488.14; found 488.15. HPLC retention time: 1.65 minutes (Method A).

N-(Benzoyl)-3-trifluoromethyl-N′-[(4-fluoro-indol-3-yl)-oxoacetyl]-piperazine (Example 143)

1

H NMR (300 MHz, CD

3

OD) δ 8.11 (s, 1H), 7.50-6.90 (m, 9H), 5.00-3.00 (m, 7H). MS m/z: (M+H)

+

calcd for C

24

H

21

F

3

N

3

O

5

: 488.14; found 488.12. HPLC retention time: 1.42 minutes (Method A).

N-(Benzoyl)-(R)-3-methyl-N′-[(4-fluoro-7-bromoindol-3-yl)-oxoacetyl]-piperazine (Example 144)

1

H NMR (CDCl

3

) δ 1.31 (br. s, 3H), 3.34 (br. s, 4H), 3.59 (br. s, 3H), 6.90 (t, J=8.7 Hz, 1H), 7.38 (br. s, 6H), 8.05 (br. s, 1H), 9.46 (br. s, 1H); MS: (M+H)

+

473.80, (M−H) 470.02; IR: 1634, 1579 cm

−1

; Anal. calcd. for C22H19N3O3BrF. 0.6H2O C, 54.68; H, 4.21; N, 8.7. Found C, 54.46; H, 4.14; N, 8.56.

1-[(Pyrid-2-yl)oxo]-4-[(1H-4-fluoro-indol-3-yl)oxoacetyl]piperazine (Example 166)

MS (ESI): 381 (M+H)

+

;

1

H NMR (CDCl

3

) δ 3.56 (m, 2H), 3.66 (m, 2H), 3.82-3.94 (m, 4H), 6.97 (m, 1H), 7.33 (m, 2H), 7.57 (m, 1H), 7.70 (m, 1H), 8.01 (m, 1H), 8.20 (s, 1H), 8.66 (m, 1H).

1-[(Pyrid-2-yl)oxo]-4-[(1H-4,7-difluoro-indol-3-yl)oxoacetyl]piperazine (Example 191)

MS (ESI): 413 (M+H)

+

;

1

H NMR (CDCl

3

) δ 3.54-3.65 (m, 4H), 3.76-3.93 (m, 4H), 6.95 (m, 2H), 7.52 (m, 1H), 7.66 (m, 1H), 7.96 (m, 1H), 8.20 (m, 1H), 8.60 (m, 1H). Example 195

MS (ESI): 370 (M+H)

+1

H NMR: (CD

3

OD, δ=3.30 ppm) 8.82 (s, 1H), 8.21 (s,1H), 8.00 (s, 1H), 7.37-7.26 (m, 2H), 7.02-6.96 (m, 1H), 3.97(b s, 2H), 3.86 (app dd, J=6.4, 3.3, 4H), 3.64 (app dd, J=6.3, 4.0, 2H); LC/MS: (ES+) m/z (M+H)

+

=370, Analytical HPLC (R

t

=0.810 min) purity: 100%. Example 212

MS (ESI): 402 (M+H)

+

1

H NMR: (CD

3

OD, δ=3.30 ppm) 8.65-8.51 (m, 2H), 8.24-8.19 (m, 1H), 8.02-7.94 (m, 1H), 7.73-7.68 (m, 2H), 7.56-7.39 (m, 2H), 4.63-3.09 (b m, 7H), 1.35 (m, 3H).

Procedures for making compounds of formula I are shown in Schemes 14-22, and further exemplified in Tables 14-18.

Starting indoles 1 (Scheme 14) are known or are readily prepared according to literature procedures, such as those described in Gribble, G. (Ref. 24) or Bartoli et al (Ref. 36). The indoles 1 are treated with oxalyl chloride in either THF (tetrahydrofuran) or ether to afford the desired glyoxyl chlorides 2 according to literature procedures (Lingens, F. et al, Ref. 25). The intermediate glyoxyl chlorides 2 are then coupled with benzoyl piperazine 3 (Desai, M. et al, Ref. 26) under basic conditions to afford 4.

Treatment of indole glyoxamide 4 (Scheme 15) with an alkylating agent (R

40

X) under basic conditions (BEMP or NaH) affords N-alkylated derivatives 5.

N-acyl derivatives 6 are prepared by treatment of indole gyloxamide 4 with an acid chloride (R

40a

COCl) in the presence of i-Pr

2

NEt (Scheme 16). Alternatively, bis-acylated products are prepared as shown in Scheme 17.

Treatment of indole-3-glyoxyl chloride 2 (Scheme 17) with tert-butyl 1-piperazinecarboxylate 7 affords the coupled product 8. Removal of the Boc protecting group of 8 is effected with 20% TFA/CH

2

Cl

2

to yield 9. This product is then coupled with acid chloride (R

40c

COCl) to afford bis-acyl products 10.

Carbamates 11 are synthesised by reaction of indole glyoxamide 4 with chloroformate (R

40d

OCOCl) in the presence of i-Pr

2

NEt or NaH (Scheme 18).

Ureas are prepared by three methods. Direct treatment of indole glyoxamide 4 with carbamoyl chloride (R

a

R

b

NCOCl) in the presence of i-Pr

2

NEt affords the desired ureas 12 (Scheme 19).

Alternatively, treatment of 4 (Scheme 20) with p-nitrophenylchloroformate and i-Pr

2

NEt affords p-nitrophenylcarbamate 13 which, on exposure to amine (R

a

R

b

NH), affords the desired urea 14.

Finally, reaction of indole glyoxamide 4 with isocyanate (R

a

NCO) in the presence of i-Pr

2

NEt affords urea 15 (Scheme 21).

Indole sulfonamides 15 (Scheme 22) are readily prepared by treatment of indole glyoxamide 4 with sulfonyl chloride (R

a

SO

2

Cl) in the presence of i-Pr

2

NEt.

TABLE 9

%

Inhibition

Example #

R

1

R

40

@ 10 μM

1

H

65

2

H

98

3

H

71

4

H

45

5

H

83

6

H

89

7

H

60

8

H

89

9

H

82

10

H

84

11

H

84

12

2-Methyl

69

13

4-Fluoro

>98

14

4-Chloro

>98

15

6-Fluoro

97

16

4,7- Difluoro

71

17

4,7- Difluoro

97

18

4,7- Difluoro

96

19

4,7- Difluoro

88

20

4,7- Difluoro

>98

21

4,7- Difluoro

93

22

4,7- Difluoro

89

23

4,7- Difluoro

98

24

4-Fluoro

91

25

4-Fluoro

92

26

4-Fluoro

95

27

4-Fluoro

98

28

4-Fluoro

96

29

4-Fluoro

93

30

4-Fluoro

92

31

4-Fluoro

82

32

4-Fluoro

97

33

4-Fluoro

80

34

4-Fluoro

TABLE 10

%

Inhibition

Example #

R

1

R

40

R

5

@ 10 μM

35

H

95

36

H

55

37

H

50

38

H

<10

39

H

81

40

H

82

41

H

86

42

H

35

43

H

86

44

H

52

45

H

64

46

H

46

47

H

>98

48

H

93

49

H

97

50

H

97

51

H

96

52

H

96

53

H

96

54

4-Chloro

>98

55

4-Fluoro

>98

56

4-Fluoro

>98

TABLE 11

Example

% Inhibition

#

R

1

R

t

@ 10 μM

57

4-Choro

Phenyl

>98

58

H

Phenyl

97

59

4-Fluoro

Phenyl

>98

60

4-Fluoro

Benzyl

>98

61

7-Methyl

t-Butyl

96

62

4-Fluoro

Methyl

>98

63

4-Fluoro

Ethyl

>98

64

4-Fluoro

t-Butyl

>98

TABLE 12

Example

% Inhibition

#

R

m

@ 10 uM

65

93

66

>98

67

97

68

97

69

>98

TABLE 13

Example

% Inhibition

#

R

n

@ 10 uM

70

Phenyl

90

71

Methyl

82

EXPERIMENTALS

3) General Procedure for Preparation of Examples 1-34

Step A.

To a solution of substituted indole IV (1 eq) in dry Et

2

O was dropwise added oxalylchloride (1.2 eq) at 0° C. After 5 min., the reaction mixture was warmed to room temperature, or heated to ˜35° C. overnight if necessary. The intermediate substituted-indole-3-glyoxyl chloride V, which was formed as a solid, was filtered and washed with dry ether (2×1 ml) to remove excessive oxalyl chloride. The product was then dried under vacuum to give desired glyoxyl chlorides V.

In cases where reaction in Et

2

O was unsuccessful, the following procedure was adopted: To a solution of substituted indole IV (1 eq) in dry THF (tetrahydrofuran) solvent was dropwise added oxalyl chloride (1.2 eq) at 0° C. After 5 min., the reaction was warmed to room temperature, or heated to ˜70° C. under nitrogen if necessary. After concentration in vacuo, the resulting crude intermediate V was submitted to next step without further treatment.

Step B

To a predried 5 ml vial was added indoles glyoxamide VIa (0.0416 μM), alkyl or aryl halide R

2

X (0.0478 μM), dry DMF (2 ml) and BEMP (0.0541 μM) at rt. The reaction was shaken at 70˜80° C. in a heating block under nitrogen for 4 hr. After evaporation of the solvent in vacuo, the crude compound was purified by prep HPLC and characterized as shown in Table 14.

For examples 33 and 34, the reactions were conducted in NMP and were heated to 80° C. for 16 h before purification by prep. HPLC.

Preparation of Example 19

To indole glyoxamide VIId (200 mg, 0.5 mmol) in THF (1 mL) in a sealed tube was added BEMP (0.2 equiv) and t-butylacrylate (0.37 mL, 2.5 mmol). The reaction mixture was heated to 90° C. overnight. The crude product was poured into 1M HCl and was extracted with EtOAc. The organic phase was washed with sat. NaCl and dried over MgSO4, filtered and concentrated. The crude product was purified by flash chromatography (2:1 EtOAc/Hexane) to afford 195 mg of alkylated product 19.

Preparation of Example 21

To ester 19 (956 mg) was added CH2Cl2 (4 mL) followed by TFA (4 mL). The reaction mixture was stirred at room temperature for 1 h. The solvent was removed in vacuo and the product was triturated with ether to afford acid 21 (802 mg) as a white solid.

Preparation of Example 22

To nitrile 17 (330 mg, 0.76 mmol) in EtOH/H2O (18 mL, 2:1) was added hydroxylamine (189 mg, 2.72 mmol) followed by K2CO3 (209 mg, 1.5 mmol). The reaction mixture was heated to 65° C. overnight. The solvent was then removed in vacuo. The residue was partitioned between water and EtOAc. The organic phase was washed with brine, dried over MgSO4, filtered and concentrated. The product was then triturated with ether to afford 22 (276 mg) as a white solid.

Preparation of Example 23

To glyoxamide 22 (100 mg, 0.21 mmol) was added toluene (1.5 mL) followed by K

2

CO

3

(35 mg, 0.26 mmol) and phosgene in toluene (1.09 mL, 20% solution). The reaction mixture was heated to reflex for 2.5 h. The mixture was then cooled to r.t. and was stirred overnight. The product was filtered, concentrated and triturated with ether to yield 23 (89 mg) as a gold colored solid.

TABLE 14

Ex-

HPCL

am-

Reten-

MS

ple

tion

Data

#

R

1

R

2

Time

(M + H)

+

1

H

1.62

402.19

2

H

1.99

400.16

3

H

1.92

520.22

4

H

1.87

466.24

5

H

1.75

482.20

6

H

1.52

376.23

7

H

1.82

432.30

8

H

1.91

458.20

9

H

1.77

430.30

10

H

1.73

390.20

11

H

1.74

418.30

12

2-Methyl

1.36

390.15

13

4-Fluoro

1.24

394.05

14

4-Chloro

1.39

410.10

15

6-Fluoro

1.38

393.79

16

4,7- Difluoro

1.16

469

17

4,7- Difluoro

1.35

437

18

4,7- Difluoro

1.49

511

19

4,7- Difluoro

1.68

526

20

4,7- Difluoro

1.47

412

21

4,7- Difluoro

1.32

470

22

4,7- Difluoro

1.18

470

23

4,7- Difluoro

1.47

496

24

4-Fluoro

1.09

451

25

4-Fluoro

1.40

442

26

4-Fluoro

2.08

494

27

4-Fluoro

2.14

522

28

4-Fluoro

2.00

466

29

4-Fluoro

1.24

471

30

4-Fluoro

1.09

471

31

4-Fluoro

1.39

471

32

4-Fluoro

1.67

494

33

4-Fluoro

2.03

528

34

4-Fluoro

1.79

501

General Procedure for Preparation of Examples 35-56

Step A

To indole-3-glyoxylyl chloride I (3 gram, 14.45 mmol) in CH

2

Cl

2

at room temperature was added tert-butyl 1-piperazinecarboxylate (2.7 gram, 14.45 mmol) and diisopropylethylamine (2.76 ml, 15.9 mmol). The light-brown color solution was stirred for 2 hr at room temperature after which time LC/MS analysis indicated the completion of the reaction. The solvent was removed in vacuo and the resulting residue was diluted with ethyl acetate (250 ml) and diethylether (250 ml). The organic solution was then washed with water (100 ml×3) and brine (50 ml), dried over MgSO

4

, filtered and concentrated. To the light-yellow solid was then added 30 ml of 20% trifluoroacetic acid in CH

2

Cl

2

. The solution was concentrated and the light-brown solid was dried in vacuo to give 3.5 g (95%) of product II. LC/MS analysis indicated this product was 100% pure and it was used for the next reaction without further purification.

Step B

To piperazine glyoxamide II (1 equiv.) in dichloroethane (DCE) was added substituted benzoyl chloride (3 equiv.) followed by i-Pr

2

NEt (4 equiv.). The reaction mixture was stirred at room temperature for 16 h and product III was then purified by prep HPLC.

General Procedure for Preparation of Examples 35-56

To indole gloxamide VIb (1 equiv.) in DCE was added substituted acid chloride (3 equiv.) followed by i-Pr

2

NEt (4 equiv.). The reaction mixture was stirred at room temperature for 16 h and product VIIa was then purified by prep HPLC.

TABLE 15

HFLC

Reten-

tion

MS

Exp. #

R

1

R

2

R

3

Time

Data

35

H

2.20

502

36

H

2.28

536

37

H

2.27

624

38

H

2.19

526

39

H

2.32

502

40

H

2.34

534

41

H

2.37

623

42

H

2.25

526

43

H

2.20

502

44

H

2.33

534

45

H

2.37

623

46

H

2.34

526

47

H

1.80

466

48

H

1.58

404

49

H

1.86

484

50

H

1.90

500

51

H

1.91

546

52

H

1.87

496

53

H

1.77

496

54

4-Chloro

1.75

500

55

4-Fluoro

1.64

484

56

4-Fluoro

1.73

514

Procedure for Preparation of Examples 57-64

To indole gloxamide VI (1 equiv.) in DCE was added chloroformate R

2

OCOCl (3 equiv.) followed by i-Pr

2

NEt (4 equiv.). The reaction mixture was stirred at room temperature for 16 h and carbamate VIIb was then purified by prep HPLC.

TABLE 16

HPLC

Example

Retention

MS Data

#

R

1

R

2

Time

(M+H)+

57

4-Choro

Phenyl

1.82

516

58

H

Phenyl

1.77

482

59

4-Fluoro

Phenyl

2.22

500

60

4-Fluoro

Benzyl

2.25

514

61

7-Methyl

t-Butyl

1.93

476

62

4-Fluoro

Methyl

1.54

438

63

4-Fluoro

Ethyl

1.65

452

64

4-Fluoro

t-Butyl

1.82

480

General Procedure for Preparation of Examples 65-69

To indole gloxamide VIe (1 equiv.) in NMP was added carbamoyl chloride R

1

R

2

NCOCl (2 equiv.) followed by i-Pr

2

NEt (4 equiv.). The reaction mixture was stirred at room temperature for 16 h and urea VIIe was then purified by prep HPLC.

General Procedure for Preparation of Examples 65-69

Step A

To indole gloxamide VIIe (1 equiv.) in DCE was added p-nitrophenylchloroformate (1.1 equiv.) followed by i-Pr

2

Nt (3 equiv.). The reaction mixture was stirred at room temperature for 3 h and the crude product was used in the following reaction without further work-up or purification.

Step B

To crude p-nitrophenylcarbamate VIIe was added secondary amine R

1

R

2

NH. The reaction mixture was stirred for 16 h at room temperature and urea IXe was then purified by prep. HPLC.

Preparation of Examples 65-69 in Table 17

To indole glyoxamide VIIe (1 equiv.) in CH

2

Cl

2

at room temperature was added isocyanate (R

1

NCO) (2 equiv.) followed by i-Pr

2

NEt (3 equiv.). The reaction mixture was stirred at room temperature for 18 h and the crude product Xe was purified by prep. HPLC.

TABLE 17

HPLC

Example

Retention

MS Data

#

R

Time

(M+H)

+

65

1.76

513

66

1.60

493

67

1.58

451

68

1.45

506

69

1.48

451

Preparation of Examples 70-71 in Table 18

To indole gloxamide XIf (1 equiv.) in DCE was added psulfonyl chloride (2 equiv.) followed by i-Pr

2

NEt (3 equiv.). The reaction mixture was stirred at room temperature for 3 h and the crude product XIIf was purified by prep. HPLC.

TABLE 18

HPLC

Example

Retention

MS Data

#

R

Time

(M+H)

+

70

Phenyl

1.85

502

71

Methyl

1.69

440

Additional Analytical Data for Selected Compounds

1

H-NMR (300 MHz,DMF-d7): δ 8.30 (s, 1H), 8.22(d, 1H, J=7.26 Hz), 7.84 (d, 1H, J=7.26 Hz), 7.30-7.54 (m, 7H), 4.99 (s, 3H), 3.52-3.92 (m, 8H).

1

H-NMR (300 MHz, CD3OD): δ 8.28 (d, 1H, J=8.37 Hz), 8.22 (s, 1H), 7.64 (d, 1H, J=7.26), 7.27-7.56 (m, 7H), 5.16 (d, 2H, J=2.52 Hz), 3.36-4.02 (m, 8H), 3.02 (t, 1H, J=2.55 Hz).

1

H-NMR (500 MHz, CD3Cl): δ 8.05 (s, 1H), 7.45 (br. s, 5H), 6.88-7.01 (m, 2H), 5.04 (s, 2H), 3.79 (br. m, 4H), 3.59 (br. m, 4H). Anal. Calcd for C

23

H

18

F

2

N

4

O

3

: C 63.30; H 4.16; N 12.84; F 8.71; O 11.00. Found C 62.33; H 4.52; N 12.05. MS 437 (M+H)

+

.

1

H-NMR (500 MHz, CD3Cl): δ 7.99 (s, 1H), 7.44 (br. s, 5H), 6.74-6.85 (m, 2H), 4.90 (s, 2H), 3.79 (br. m, 4H), 3.57 (br. m, 4H), 3.45 (q, J=11.4 Hz, 4H). 1.35 (t, J=11.4 Hz, 3H), 1.21 (t, J=11.4 Hz, 3H). Anal. Calcd for C

27

H

28

F

2

N

4

O

4

: C 63.52; H 5.54; N 10.97. Found C 62.75; H 5.54; N 10.80.

1

H-NMR (500 MHz, DMSO-d6): δ 8.32 (s, 1H), 7.44 (br. s, 6H), 7.12 (app. t, 1H), 3.87 (s, 3H), 3.2-3.7 (br. m, 8H). MS 412 (M+H)

+

Anal. for C

22

H

19

F

2

N

3

O

3

: C 64.23; H 4.65; N 10.1. Found C

64.58

; H 4.74; N 9.63.

1

H-NMR (500 MHz, DMSO-d6): δ 8.40 (s, 1H), 7.44 (br. s, 6H), 7.14 (app. t, 1H), 5.01 (br. s, 2H), 3.2-3.7 (br. m). MS 470 (M+H)

+

Anal. for C

23

H

21

F

2

N

5

O

4

: C 58.85; H 4.51; N 14.92. Found C 58.17; H 5.06; N 13.85.

1

H-NMR (500 MHz, DMSO-d6): δ 8.40 (s, 1H), 7.4-7.55 (m, 6H), 7.16 (app. t, 1H), 5.61 (br. s, 2H), 3.2-3.7 (br. m). MS 496 (M+H)

+

The compounds of the present invention may be administered orally, parenterally (including subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques), by inhalation spray, or rectally, in dosage unit formulations containing conventional non-toxic pharmaceutically-acceptable carriers, adjuvants and vehicles.

Thus, in accordance with the present invention there is further provided a method of treating and a pharmaceutical composition for treating viral infections such as HIV infection and AIDS. The treatment involves administering to a patient in need of such treatment a pharmaceutical composition comprising a pharmaceutical carrier and a therapeutically-effective amount of a compound of the present invention.

The pharmaceutical composition may be in the form of orally-administrable suspensions or tablets; nasal sprays, sterile injectable preparations, for example, as sterile injectable aqueous or oleagenous suspensions or suppositories.

When administered orally as a suspension, these compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may contain microcrystalline cellulose for imparting bulk, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, and sweetners/flavoring agents known in the art. As immediate release tablets, these compositions may contain microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate and lactose and/or other excipients, binders, extenders, disintegrants, diluents and lubricants known in the art.

The injectable solutions or suspensions may be formulated according to known art, using suitable non-toxic, parenterally-acceptable diluents or solvents, such as mannitol, 1,3-butanediol, water, Ringer's solution or isotonic sodium chloride solution, or suitable dispersing or wetting and suspending agents, such as sterile, bland, fixed oils, including synthetic mono- or diglycerides, and fatty acids, including oleic acid.

The compounds of this invention can be administered orally to humans in a dosage range of 1 to 100 mg/kg body weight in divided doses. One preferred dosage range is 1 to 10 mg/kg body weight orally in divided doses. Another preferred dosage range is 1 to 20 mg/kg body weight orally in divided doses. It will be understood, however, that the specific dose level and frequency of dosage for any particular patient 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 health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy.

Abbreviations

TFA

Trifluoroacetic Acid

P-EDC

Polymer supported 1-(3-dimethylaminopropyl)-3-

ethylcarbodiimide

EDC

1-(3-dimethylaminopropyl)-3-ethylcarbodiimide

DCE

1,2-Dichloroethane

DMF

N,N-dimethylformamide

THF

Tetrahydrofuran

NMP

N-methylpyrrolidone

BEMP

2-tert-Butylimino-2-diethylamino-1,3-dimethyl-

perhydro-1,3,2-diazaphosphorine

Number	Name	Date	Kind
5124327	Greenlee et al.	Jun 1992	A
5192770	Clark et al.	Mar 1993	A
5424329	Boschelli et al.	Jun 1995	A
5681954	Yamamoto et al.	Oct 1997	A

Number	Date	Country
0484071	May 1992	EP
0530907	Mar 1993	EP
WO 9301181	Jan 1993	WO
9709308	Mar 1997	WO
WO 9955696	Nov 1999	WO

Antiviral indoleoxoacetyl piperazine derivatives

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CPC

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