U.S. patent application number 10/871931 was filed with the patent office on 2005-04-07 for indole, azaindole and related heterocyclic n-substituted piperazine derivatives.
Invention is credited to Coulter, Thomas Stephen, Farkas, Michelle, Johnston, David, Kadow, John F., Meanwell, Nicholas A., Taylor, Malcolm, Wright, J. J. Kim, Yeung, Kap-Sun.
Application Number | 20050075364 10/871931 |
Document ID | / |
Family ID | 34069101 |
Filed Date | 2005-04-07 |
United States Patent
Application |
20050075364 |
Kind Code |
A1 |
Yeung, Kap-Sun ; et
al. |
April 7, 2005 |
Indole, azaindole and related heterocyclic N-substituted piperazine
derivatives
Abstract
This invention provides compounds of Formula I, including
pharmaceutically accceptable salts thereof, having drug and
bio-affecting properties, their pharmaceutical compositions and
method of use. 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. The compounds of Formula I have the formula 1 wherein: Z
is 2 Q is selected from the group consisting of 3 m is 2; A is
selected from the group consisting of cinnolinyl, napthyridinyl,
quinoxalinyl, pyridinyl, pyrimidinyl, quinolinyl, isoquinolinyl,
quinazolinyl, azabenzofuryl, and phthalazinyl each of which may be
optionally substituted with one or two groups independently
selected from methyl, methoxy, hydroxy, amino and halogen; and
--W-- is 4
Inventors: |
Yeung, Kap-Sun; (Madison,
CT) ; Farkas, Michelle; (Pasadena, CA) ;
Kadow, John F.; (Wallingford, CT) ; Meanwell,
Nicholas A.; (East Hampton, CT) ; Taylor,
Malcolm; (Didcot, GB) ; Johnston, David;
(Didcot, GB) ; Coulter, Thomas Stephen; (Wantage,
GB) ; Wright, J. J. Kim; (Redwood City, CA) |
Correspondence
Address: |
STEPHEN B. DAVIS
BRISTOL-MYERS SQUIBB COMPANY
PATENT DEPARTMENT
P O BOX 4000
PRINCETON
NJ
08543-4000
US
|
Family ID: |
34069101 |
Appl. No.: |
10/871931 |
Filed: |
June 18, 2004 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
60541970 |
Feb 5, 2004 |
|
|
|
60493283 |
Aug 7, 2003 |
|
|
|
60484224 |
Jul 1, 2003 |
|
|
|
Current U.S.
Class: |
514/300 ;
514/419; 546/113; 548/495 |
Current CPC
Class: |
A61P 31/00 20180101;
C07D 491/04 20130101; C07D 403/12 20130101; C07D 413/14 20130101;
C07D 401/12 20130101; A61P 37/02 20180101; C07D 471/04 20130101;
A61P 31/18 20180101; C07D 401/14 20130101 |
Class at
Publication: |
514/300 ;
514/419; 546/113; 548/495 |
International
Class: |
C07D 471/02; A61K
031/4745; A61K 031/405 |
Claims
What is claimed is:
1. A compound of Formula I, including pharmaceutically acceptable
salts thereof, 465wherein: Z is 466Q is selected from the group
consisting of 467R.sup.1 is hydrogen; R.sup.2, R.sup.3, R.sup.4,
and R.sup.5, are independently selected from the group consisting
of hydrogen, halogen, cyano, COOR.sup.8, XR.sup.9 and B; m is 2;
R.sup.6 is 0 or does not exist; R.sup.7 is hydrogen; R.sup.10 is
selected from the group consisting of (C.sub.1-6)alkyl,
--CH.sub.2CN, --CH.sub.2COOH, --CH.sub.2C(O)NR.sup.11R.sup.12,
phenyl and pyridinyl; R.sup.11 and R.sup.12 are each independently
H or (C.sub.1-3)alkyl; - - represents a carbon-carbon bond; A is
selected from the group consisting of cinnolinyl, napthyridinyl,
quinoxalinyl, pyridinyl, pyrimidinyl, quinolinyl, isoquinolinyl,
quinazolinyl, azabenzofuryl, and phthalazinyl each of which may be
optionally substituted with one or two groups independently
selected from methyl, methoxy, hydroxy, amino and halogen; --W-- is
468R.sup.15, R.sup.16, R.sup.17, R.sup.18, R.sup.19, R.sup.20,
R.sup.21, R.sup.22 are each independently H or one of them is
methyl; B is selected from the group consisting of
C(O)NR.sup.11R.sup.12C(.dbd.NH)N- HNHC(O)--R.sup.10,
C(.dbd.NH)cyclopropyl, C(.dbd.NOH)NH.sub.2, and heteroaryl; wherein
said heteroaryl is independently optionally substituted with a
substituent selected from F; heteroaryl is selected from the group
consisting of pyridinyl, pyrazinyl, pyrimidinyl, thiazolyl,
pyrrolyl, imidazolyl, benzoimidazolyl, oxadiazolyl, pyrazolyl,
tetrazolyl and triazolyl; F is selected from the group consisting
of (C.sub.1-6)alkyl, (C.sub.1-6)alkoxy, cyano,
COOR.sup.8--CONR.sup.11R.sup.- 12; --CH.sub.2CN, --CH.sub.2COOH,
--CH.sub.2C(O)NR.sup.11R.sup.12, phenyl and pyridinyl; R.sup.8 and
R.sup.9 are independently selected from the group consisting of
hydrogen and (C.sub.1-6)alkyl; X is O; provided that when A is
pyridinyl or pyrimidinyl and Q is 469 then R.sup.5 is B.
2. A compound of claim 1, wherein: R.sup.15, R.sup.16, R.sup.17,
R.sup.18, R.sup.19, R.sup.20, R.sup.21, R.sup.22 are H; R.sup.6
does not exist; A is selected from members of the group consisting
of 470 where Xw is the point of attachment and each member is
independently optionally substituted with one group selected from
the group consisting of methyl, methoxy, hydroxy, amino and
halogen; Q is selected from the group consisting of 471 provided
when Q is 472 then R.sup.2 is hydrogen, methoxy or halogen; R.sup.3
and R.sup.4 are hydrogen; and R.sup.5 is selected from the group
consisting of hydrogen, halogen, cyano, COOR.sup.8, XR.sup.9 and B;
or provided when Q is 473 then R.sup.2 is hydrogen, methoxy or
halogen; R.sup.3 is hydrogen; and R.sup.4 is selected from the
group consisting of hydrogen, halogen, cyano, COOR.sup.8, XR.sup.9
and B; or provided when Q is 474 then R.sup.2 and R.sup.3 are each
hydrogen; and R.sup.4 is selected from the group consisting of
hydrogen, halogen, cyano, COOR.sup.8, XR.sup.9 and B.
3. A compound of claim 2 wherein: B is selected from the group
consisting of C(O)NR.sup.11R.sup.12 and heteroaryl; wherein said
heteroaryl is independently optionally substituted with a
substituent selected from F; heteroaryl is selected from the group
consisting of pyridinyl, pyrazinyl, pyrimidinyl, thiazolyl,
pyrrolyl, imidazolyl, benzoimidazolyl, oxadiazolyl, tetrazolyl and
triazolyl.
4. A compound of claim 3 wherein: B is heteroaryl wherein said
heteroaryl is independently optionally substituted with a
substituent selected from F.
5. A compound of claim 3 wherein: A is selected from the group
consisting of 475 where Xw is the point of attachment.
6. A compound of claim 5 wherein: B is heteroaryl; wherein said
heteroaryl is independently optionally substituted with a
substituent selected from F; and heteroaryl is selected from the
group consisting of triazolyl, pyridinyl, pyrazinyl and
pyrimidinyl.
7. A compound of claim 6 wherein: B is heteroaryl; wherein said
heteroaryl is independently optionally substituted with a
substituent selected from F; and heteroaryl is selected from the
group consisting of triazolyl.
8. A compound of claim 7 where F is methyl.
9. A pharmaceutical composition which comprises an antiviral
effective amount of a compound of Formula I, including
pharmaceutically acceptable salts thereof, as claimed in claim 1,
and one or more pharmaceutically acceptable carriers, excipients or
diluents.
10. The pharmaceutical composition of claim 9, useful for treating
infection by 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.
11. A method for treating a mammal infected with the HIV virus
comprising administering to said mammal an antiviral effective
amount of a compound of Formula I, including pharmaceutically
accceptable salts thereof, as claimed in claim 1, and one or more
pharmaceutically acceptable carriers, excipients or diluents.
12. The method of claim 11, comprising administering to said mammal
an antiviral effective amount of a compound of Formula I, including
pharmaceutically accceptable salts thereof, in combination with an
antiviral effective amount of an AIDS treatment agent selected from
the group consisting of an AIDS antiviral agent; an anti-infective
agent; an immunomodulator; and an HIV entry inhibitor.
Description
REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Application Ser. No. 60/541,970 filed Feb. 5, 2004, 60/493,283
filed Aug. 7, 2003 and 60/484,224 filed Jul. 1, 2003.
FIELD OF THE INVENTION
[0002] This invention provides compounds having drug and
bio-affecting properties, their pharmaceutical compositions and
method of use. In particular, the invention is concerned with new
N-heteroaryl and N-aryl piperazine derivatives that possess unique
antiviral activity. More particularly, the present invention
relates to compounds useful for the treatment of HIV and AIDS.
BACKGROUND ART
[0003] HIV-1 (human immunodeficiency virus-1) infection remains a
major medical problem, with an estimated 42 million people infected
worldwide at the end of 2002. The number of cases of HIV and AIDS
(acquired immunodeficiency syndrome) has risen rapidly. In 2002,
.about.5.0 million new infections were reported, and 3.1 million
people died from AIDS. Currently available drugs for the treatment
of HIV include ten nucleoside reverse transcriptase (RT) inhibitors
or approved single pill combinations(zidovudine or AZT (or
Retrovir.RTM.), didanosine (or Videx.RTM.), stavudine (or
Zerit.RTM.), lamivudine (or 3TC or Epivir.RTM.), zalcitabine (or
DDC or Hivid.RTM.), abacavir succinate (or Ziagen.RTM.), Tenofovir
disoproxil fumarate salt (or Viread.RTM.), Combivir.RTM. (contains
-3TC plus AZT), Trizivir.RTM. (contains abacavir, lamivudine, and
zidovudine), Emtriva.RTM. (emtricitabine); three non-nucleoside
reverse transcriptase inhibitors: nevirapine (or Viramune.RTM.),
delavirdine (or Rescriptor.RTM.) and efavirenz (or Sustiva.RTM.),
and eight peptidomimetic protease inhibitors or approved
formulations: saquinavir, indinavir, ritonavir, nelfinavir,
amprenavir, lopinavir, Kaletra.RTM. (lopinavir and Ritonavir), and
Reyataz.RTM. (atazanavir). 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 viremia and
disease progression. In fact, significant reductions in death rates
among AIDS patients have been recently documented as a consequence
of the widespread application of combination therapy. However,
despite these impressive 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 sub-optimal
drug concentrations are present (Larder and Kemp; Gulick;
Kuritzkes; Morris-Jones et al; Schinazi et al; Vacca and Condra;
Flexner; Berkhout and Ren et al; (Ref. 6-14)). Therefore, novel
anti-HIV agents exhibiting distinct resistance patterns, and
favorable pharmacokinetic as well as safety profiles are needed to
provide more treatment options.
[0004] Currently marketed HIV-1 drugs are dominated by either
nucleoside reverse transcriptase inhibitors or peptidomimetic
protease inhibitors. Non-nucleoside reverse transcriptase
inhibitors (NNRTIs) have recently gained an increasingly important
role in the therapy of HIV infections (Pedersen & Pedersen, Ref
15). At least 30 different classes of NNRTI have been described in
the literature (De Clercq, Ref. 16) and several NNRTIs have been
evaluated in clinical trials. Dipyridodiazepinone (nevirapine),
benzoxazinone (efavirenz) and bis(heteroaryl) piperazine
derivatives (delavirdine) have been approved for clinical use.
However, the major drawback to the development and application of
NNRTIs is the propensity for rapid emergence of drug resistant
strains, both in tissue cell culture and in treated individuals,
particularly those subject to monotherapy. As a consequence, there
is considerable interest in the identification of NNRTIs less prone
to the development of resistance (Pedersen & Pedersen, Ref 15).
A recent overview of non-nucleoside reverse transcriptase
inhibitors: perspectives on novel therapeutic compounds and
strategies for the treatment of HIV infection. has appeared
(Buckheit, reference 99). A review covering both NRTI and NNRTIs
has appeared (De clercq, reference 100). An overview of the current
state of the HIV drugs has been published (De clercq, reference
101).
[0005] 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. 17;
Romero et al, Ref. 18; Young et al, Ref. 19; Genin et al, Ref. 20;
Silvestri et al, Ref. 21). Indole 2-carboxamides have also been
described as inhibitors of cell adhesion and HIV infection
(Boschelli et al, U.S. Pat. No. 5,424,329, Ref. 4). 3-substituted
indole natural products (Semicochliodinol A and B,
didemethylasterriquinone and isocochliodinol) were disclosed as
inhibitors of HIV-1 protease (Fredenhagen et al, Ref. 22).
[0006] Structurally related aza-indole amide derivatives have been
disclosed previously (Kato et al, Ref. 23; Levacher et al, Ref. 24;
Dompe Spa, WO-09504742, Ref. 5(a); SmithKline Beecham PLC,
WO-09611929, Ref. 5(b); Schering Corp., U.S.-05023265, Ref. 5(c)).
However, these structures differ from those claimed herein in that
they are aza-indole mono-amide rather than oxoacetamide
derivatives, and there is no mention of the use of these compounds
for treating viral infections, particularly HIV. PCT International
Patent Application WO9951224 by Bernd Nickel et. al. (reference
107) describes N-indolylglyoxamides for the treatment of cancer.
Although some of these compounds contain N-heteroaryl or N-aryl
piperazines, the substitution patterns at the other positions are
outside the scope of this invention.
[0007] The compounds of this invention inhibit HIV entry by
attaching to the exterior viral envelop protein gp120 and
interrupting the viral entry process, possibly by interfering with
recognition of the cellular receptor CD4. Compounds in this class
have been reported to have antiviral activity against a variety of
laboratory and clinical strains of HIV-1 and are effective in
treating HIV infection (see Hanna et al., Abstract 141 presented at
the 11th Conference on Retroviruses and Opportunistic Infections,
San Francisco, Calif., Feb. 8-11, 2004; Lin et al., Poster 534
presented at the 11th Conference on Retroviruses and Opportunistic
Infections, San Francisco, Calif., Feb. 8-11, 2004; Hanna et al.,
Poster 535 presented at the 11th Conference on Retroviruses and
Opportunistic Infections, San Francisco, Calif., Feb. 8-11,
2004).
[0008] N-(3-aryl-3-oxo)acetyl piperidines have been disclosed. See
Blair et al., U.S. Pat. No. 6,469,006; Wang et al., U.S. Pat. No.
6,476,034; Wang et al., U.S. Pat. No. 6,632,819; Wallace et al.,
U.S. Pat. No. 6,573,262 (continuation-in-part application of U.S.
Ser. No. 09/888,686, filed Jun. 25, 2001); Wang et al., U.S. patent
application Ser. No. 10/214,982 filed Aug. 7, 2002
(continuation-in-part application of U.S. Ser. No. 10/038,306 filed
Jan. 2, 2002); Wang et al., patent application WO 03/092695,
published Nov. 13, 2003; Kadow et. al. patent application WO
04/000210 published Dec. 31, 2003; Regueiro-Ren et. al. patent
application WO 04/011425 published Feb. 5, 2004; Wang et al., US
patent application US 20040063744, published Apr. 1, 2004. Nothing
in these references teaches or suggests the novel compounds of this
invention or their use to inhibit HIV infection.
REFERENCES CITED
[0009] Patent Documents
[0010] 1. Greenlee, W. J.; Srinivasan, P. C. Indole reverse
transcriptase inhibitors. U.S. Pat. No. 5,124,327.
[0011] 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.
[0012] 3. Romero, D. L.; Thomas, R. C.; Preparation of substituted
indoles as anti-AIDS pharmaceuticals. PCT WO 93/01181.
[0013] 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.
[0014] 5. (a) Mantovanini, M.; Melillo, G.; Daffonchio, L. Tropyl
7-azaindol-3-ylcarboxyamides as antitussive agents. PCT WO 95/04742
(Dompe Spa). (b) Cassidy, F.; Hughes, I.; Rahman, S.; Hunter, D. J.
Bisheteroaryl-carbonyl and carboxamide derivatives with 5HT 2C/2B
antagonists activity. PCT WO 96/11929.
[0015] (c) Scherlock, M. H.; Tom, W. C. Substituted
1H-pyrrolopyridine-3-carboxamides. U.S. Pat. No. 5,023,265.
[0016] Other Publications
[0017] 6. Larder, B. A.; Kemp, S. D. Multiple mutations in the
HIV-1 reverse transcriptase confer high-level resistance to
zidovudine (AZT). Science, 1989, 246,1155-1158.
[0018] 7. Gulick, R. M. Current antiretroviral therapy: An
overview. Quality of Life Research, 1997, 6, 471-474.
[0019] 8. Kuritzkes, D. R. HIV resistance to current therapies.
Antiviral Therapy, 1997, 2 (Supplement 3), 61-67.
[0020] 9. Morris-Jones, S.; Moyle, G.; Easterbrook, P. J.
Antiretroviral therapies in HIV-1 infection. Expert Opinion on
Investigational Drugs, 1997, 6(8), 1049-1061.
[0021] 10. Schinazi, R. F.; Larder, B. A.; Mellors, J. W. Mutations
in retroviral genes associated with drug resistance. International
Antiviral News, 1997, 5,129-142.
[0022] 11. Vacca, J. P.; Condra, J. H. Clinically effective HIV-1
protease inhibitors. Drug Discovery Today, 1997, 2, 261-272.
[0023] 12. Flexner, D. HIV-protease inhibitors. Drug Therapy, 1998,
338, 1281-1292.
[0024] 13. Berkhout, B. HIV-1 evolution under pressure of protease
inhibitors: Climbing the stairs of viral fitness. J. Biomed. Sci.,
1999, 6, 298-305.
[0025] 14. Ren, S.; Lien, E. J. Development of HIV protease
inhibitors: A survey. Prog. Drug Res., 1998, 51, 1-31.
[0026] 15. Pedersen, O. S.; Pedersen, E. B. Non-nucleoside reverse
transcriptase inhibitors: the NNRTI boom. Antiviral Chem.
Chemother. 1999, 10, 285-314.
[0027] 16. (a) De Clercq, E. The role of non-nucleoside reverse
transcriptase inhibitors (NNRTIs) in the therapy of HIV-1
infection. Antiviral Research, 1998, 38, 153-179. (b) De Clercq, E.
Perspectives of non-nucleoside reverse transcriptase inhibitors
(NNRTIs) in the therapy of HIV infection. IL. Farmaco, 1999, 54,
26-45.
[0028] 17. Font, M.; Monge, A.; Cuartero, A.; Elorriaga, A.;
Martinez-lrujo, J. J.; Alberdi, E.; Santiago, E.; Prieto, I.;
Lasarte, J. J.; Sarobe, P. and Borras, F. Indoles and
pyrazino[4,5-b]indoles as normucleoside analog inhibitors of HIV-1
reverse transcriptase. Eur. J. Med. Chem., 1995, 30, 963-971.
[0029] 18. 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)-carbony-
l]-4-[3-[1-methylethyl)amino]-pyridinyl]piperazine
momomethansulfonate (U-90152S), a second generation clinical
candidate. J. Med. Chem., 1993, 36, 1505-1508.
[0030] 19. 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., 1995, 5, 491-496.
[0031] 20. 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., 1996, 39,
5267-5275.
[0032] 21. 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]benzothiazepi- ne derivatives, designed as
conformationally constrained analogues of the human
immunodeficiency virus type 1 reverse transcriptase inhibitor
L-737,126. Antiviral Chem. Chemother. 1998, 9, 139-148.
[0033] 22. 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, 1997, 50, 395-401.
[0034] 23. Kato, M.; Ito, K.; Nishino, S.; Yamakuni, H.; Takasugi,
H. New 5-HT.sub.3 (Serotonin-3) receptor antagonists. IV. Synthesis
and structure-activity relationships of azabicycloalkaneacetamide
derivatives. Chem. Pharm. Bull., 1995, 43, 1351-1357.
[0035] 24. Levacher, V.; Benoit, R.; Duflos, J; Dupas, G.;
Bourguignon, J.; Queguiner, G. Broadening the scope of NADH models
by using chiral and non chiral pyrrolo [2,3-b]pyridine derivatives.
Tetrahedron, 1991, 47, 429-440.
[0036] 25. Shadrina, L. P.; Dormidontov, Yu. P.; Ponomarev, V, G.;
Lapkin, I. I. Reactions of organomagnesium derivatives of 7-aza-
and benzoindoles with diethyl oxalate and the reactivity of
ethoxalylindoles. Khim. Geterotsikl. Soedin., 1987, 1206-1209.
[0037] 26. Sycheva, T. V.; Rubtsov, N. M.; Sheinker, Yu. N.;
Yakhontov, L. N. Some reactions of 5-cyano-6-chloro-7-azaindoles
and lactam-lactim tautomerism in 5-cyano-6-hydroxy-7-azaindolines.
Khim. Geterotsikl. Soedin., 1987, 100-106.
[0038] 27. (a) Desai, M.; Watthey, J. W. H.; Zuckerman, M. A
convenient preparation of 1-aroylpiperazines. Org. Prep. Proced.
Int., 1976, 8, 85-86. (b) Adamczyk, M.; Fino, J. R. Synthesis of
procainamide metabolites. N-acetyl desethylprocainamide and
desethylprocainamide. Org. Prep. Proced. Int. 1996, 28, 470-474.
(c) 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. (d) Wang, T.; Zhang, Z.; Meanwell, N. A. Benzoylation of
Dianions: Preparation of mono-Benzoylated Symmetric Secondary
Diamines. J. Org. Chem., 1999, 64, 7661-7662.
[0039] 28. Li, H.; Jiang, X.; Ye, Y.-H.; Fan, C.; Romoff, T.;
Goodman, M. 3-(Diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one
(DEPBT): A new coupling reagent with remarkable resistance to
racemization. Organic Lett., 1999, 1, 91-93.
[0040] 29. Harada, N.; Kawaguchi, T.; Inoue, I.; Ohashi, M.; Oda,
K.; Hashiyama, T.; Tsujihara, K. Synthesis and antitumor activity
of quaternary salts of 2-(2'-oxoalkoxy)-9-hydroxyellipticines.
Chem. Pharm. Bull., 1997, 45, 134-137.
[0041] 30. Schneller, S. W.; Luo, J.-K. Synthesis of
4-amino-1H-pyrrolo[2,3-b]pyridine (1,7-Dideazaadenine) and
1H-pyrrolo[2,3-b]pyridin-4-ol (1,7-Dideazahypoxanthine). J. Org.
Chem., 1980, 45, 4045-4048.
[0042] 31. Shiotani, S.; Tanigochi, K. Furopyridines. XXII [1].
Elaboration of the C-substitutents alpha to the heteronitrogen atom
of furo[2,3-b]-, -[3.2-b]-, -[2.3-c]- and -[3,2-c]pyridine. J. Het.
Chem., 1997, 34, 901-907.
[0043] 32. Minakata, S.; Komatsu, M.; Ohshiro, Y. Regioselective
functionalization of 1H-pyrrolo[2,3-b]pyridine via its N-oxide.
Synthesis, 1992, 661-663.
[0044] 33. Klemm, L. H.; Hartling, R. Chemistry of thienopyridines.
XXIV. Two transformations of thieno[2,3-b]pyridine 7-oxide (1). J.
Het. Chem., 1976, 13, 1197-1200.
[0045] 34. Antonini, I.; Claudi, F.; Cristalli, G.; Franchetti, P.;
Crifantini, M.; Martelli, S. Synthesis of
4-amino-1-.beta.-D-ribofuranosy- l-1H-pyrrolo[2,3-b]pyridine
(1-Deazatubercidin) as a potential antitumor agent. J. Med. Chem.,
1982, 25, 1258-1261.
[0046] 35. (a) Regnouf De Vains, J. B.; Papet, A. L.; Marsura, A.
New symmetric and unsymmetric polyfunctionalized 2,2'-bipyridines.
J. Het. Chem., 1994, 31, 1069-1077. (b) Miura, Y.; Yoshida, M.;
Hamana, M. Synthesis of 2,3-fused quinolines from 3-substituted
quinoline 1-oxides. Part II, Heterocycles, 1993, 36, 1005-1016. (c)
Profft, V. E.; Rolle, W. Uber 4-merkaptoverbindungendes
2-methylpyridins. J. Prakt. Chem., 1960, 283 (11), 22-34.
[0047] 36. Nesi, R.; Giomi, D.; Turchi, S.; Tedeschi, P.,
Ponticelli, F. A new one step synthetic approach to the
isoxazolo[4,5-b]pyridine system. Synth. Comm., 1992, 22,
2349-2355.
[0048] 37. (a) Walser, A.; Zenchoff, G.; Fryer, R. I. Quinazolines
and 1,4-benzodiazepines. 75. 7-Hydroxyaminobenzodiazepines and
derivatives. J. Med. Chem., 1976, 19, 1378-1381. (b) Barker, G.;
Ellis, G. P. Benzopyrone. Part I. 6-Amino- and
6-hydroxy-2-subtituted chromones. J. Chem. Soc., 1970,
2230-2233.
[0049] 38. Ayyangar, N. R.; Lahoti, R J.; Daniel, T. An alternate
synthesis of 3,4-diaminobenzophenone and mebendazole. Org. Prep.
Proced. Int., 1991, 23, 627-631.
[0050] 39. Mahadevan, I.; Rasmussen, M. Ambident heterocyclic
reactivity: The alkylation of pyrrolopyridines (azaindoles,
diazaindenes). Tetrahedron, 1993, 49, 7337-7352.
[0051] 40. Chen, B. K.; Saksela, K.; Andino, R.; Baltimore, D.
Distinct modes of human immunodeficiency type 1 proviral latency
revealed by superinfection of nonproductively infected cell lines
with recombinant luciferase-encoding viruses. J. Virol., 1994, 68,
654-660.
[0052] 41. Bodanszky, M.; Bodanszky, A. "The Practice of Peptide
Synthesis" 2.sup.nd Ed., Springer-Verlag: Berlin Heidelberg,
Germany, 1994.
[0053] 42. Albericio, F. et al. J. Org. Chem. 1998, 63, 9678.
[0054] 43. Knorr, R. et al. Tetrahedron Lett. 1989, 30, 1927.
[0055] 44. (a) Jaszay Z. M. et al. Synth. Commun., 1998 28, 2761
and references cited therein; (b) Bemasconi, S. et al. Synthesis,
1980, 385.
[0056] 45. (a) Jaszay Z. M. et al. Synthesis, 1989, 745 and
references cited therein; (b) Nicolaou, K. C. et al. Angew. Chem.
Int. Ed. 1999, 38, 1669.
[0057] 46. Ooi, T. et al. Synlett. 1999, 729.
[0058] 47. Ford, R. E. et al. J. Med. Chem. 1986, 29, 538.
[0059] 48. (a) Yeung, K.-S. et al. Bristol-Myers Squibb Unpublished
Results. (b) Wang, W. et al. Tetrahedron Lett. 1999, 40, 2501.
[0060] 49. Brook, M. A. et al. Synthesis, 1983, 201.
[0061] 50. Yamazaki, N. et al. Tetrahedron Lett. 1972, 5047.
[0062] 51. Barry A. Bunin "The Combinatorial Index" 1998 Academic
Press, San Diego/London pages 78-82.
[0063] 52. Richard C. Larock Comprehensive Organic Transormations
2nd Ed. 1999, John Wiley and Sons New York.
[0064] 53. M. D. Mullican et. al. J. Med. Chem. 1991, 34,
2186-2194.
[0065] 54. Protective groups in organic synthesis 3rd ed./Theodora
W. Greene and Peter G. M. Wuts. New York: Wiley, 1999.
[0066] 55. Katritzky, Alan R. Lagowski, Jeanne M. The principles of
heterocyclic Chemistry New York: Academic Press, 1968.
[0067] 56. Paquette, Leo A. Principles of modern heterocyclic
chemistry New York: Benjamin.
[0068] 57. Katritzky, Alan R.; Rees, Charles W.; Comprehensive
heterocyclic chemistry: the structure, reactions, synthesis, and
uses of heterocyclic compounds 1st ed. Oxford (Oxfordshire); New
York: Pergamon Press, 1984. 8 v.
[0069] 58. Katritzky, Alan RHandbook of heterocyclic 1st ed Oxford
(Oxfordshire); New York: Pergamon Press, 1985.
[0070] 59. Davies, David I Aromatic Heterocyclic Oxford; New York:
Oxford University Press, 1991.
[0071] 60. Ellis, G. P. Synthesis of fused Chichester [Sussex]; New
York: Wiley, c1987-c1992. Chemistry of heterocyclic compounds; v.
47.
[0072] 61. Joule, J. A Mills, K., Smith, G. F. Heterocyclic
Chemistry, 3rd ed London; New York Chapman & Hall, 1995.
[0073] 62. Katritzky, Alan R., Rees, Charles W., Scriven, Eric F.
V. Comprehensive heterocyclic chemistry II: a review of the
literature 1982-1995.
[0074] 63. The structure, reactions, synthesis, and uses of
heterocyclic compounds 1st ed. Oxford; New York: Pergamon, 1996. 11
v. in 12: ill.; 28 cm.
[0075] 64. Eicher, Theophil, Hauptmann, Siegfried. The chemistry of
heterocycles: structure, reactions, syntheses, and applications
Stuttgart; New York: G. Thieme, 1995.
[0076] 65. Grimmett, M. R. Imidazole and benzimidazole Synthesis
London; San Diego: Academic Press, 1997.
[0077] 66. Advances in heterocyclic chemistry. Published in New
York by Academic Press, starting in 1963-present.
[0078] 67. Gilchrist, T. L. (Thomas Lonsdale) Heterocyclic
chemistry 3rd ed. Harlow, Essex: Longman, 1997, 414 p: ill.; 24
cm.
[0079] 68. Farina, Vittorio; Roth, Gregory P. Recent advances in
the Stille reaction; Adv. Met.-Org. Chem. 1996, 5, 1-53.
[0080] 69. Farina, Vittorio; Krishnamurthy, Venkat; Scott, William
J. The Stille reaction; Org. React. (N.Y.) (1997), 50, 1-652.
[0081] 70. Stille, J. K. Angew. Chem. Int. Ed. Engl. 1986, 25,
508-524.
[0082] 71. Norio Miyaura and Akiro Suzuki Chem Rev. 1995, 95,
2457.
[0083] 72. Home, D. A. Heterocycles 1994, 39, 139.
[0084] 73. Kamitori, Y. et. al. Heterocycles, 1994, 37(1), 153.
[0085] 74. Shawali, J. Heterocyclic Chem. 1976, 13, 989.
[0086] 75. a) Kende, A. S. et al. Org. Photochem. Synth. 1972, 1,
92. b) Hankes, L. V.; Biochem. Prep. 1966, 11, 63. c) Synth. Meth.
22, 837.
[0087] 76. Hulton et. al. Synth. Comm. 1979, 9, 789.
[0088] 77. Pattanayak, B. K. et. al. Indian J. Chem. 1978, 16,
1030.
[0089] 78. Chemische Berichte 1902, 35, 1545.
[0090] 79. Chemische Berichte Ibid 1911, 44, 493.
[0091] 80. Moubarak, I., Vessiere, R. Synthesis 1980, Vol. 1,
52-53.
[0092] 81. Ind J. Chem. 1973, 11, 1260.
[0093] 82. Roomi et. al. Can J. Chem. 1970, 48, 1689.
[0094] 83. Sorrel, T. N. J. Org. Chem. 1994, 59, 1589.
[0095] 84. Nitz, T. J. et. al. J. Org. Chem. 1994, 59,
5828-5832.
[0096] 85. Bowden, K. et. al. J. Chem. Soc. 1946, 953.
[0097] 86. Nitz, T. J. et. al. J. Org. Chem. 1994, 59,
5828-5832.
[0098] 87. Scholkopf et. al. Angew. Int. Ed. Engl. 1971, 10(5),
333.
[0099] 88. (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. Recl. Trav. Chim. Pays-Bas
1995,114, 97.
[0100] 89. Wang, T.; Zhang, Z.; Meanwell, N. A. Benzoylation of
Dianions: Preparation of mono-Benzoylated Symmetric Secondary
Diamines. J. Org. Chem., 1999, 64, 7661-7662.
[0101] 90. (a) Adamczyk, M.; Fino, J. R. Synthesis of procainamide
metabolites. N-acetyl desethylprocainamide and
desethylprocainamide. Org. Prep. Proced. Int. 1996, 28, 470-474.
(b) Wang, T.; Zhang, Z.; Meanwell, N. A. Regioselective
mono-Benzoylation of Unsymmetrical piperazines. J. Org. Chem., in
press.
[0102] 91. Masuzawa, K.; Kitagawa, M.; Uchida, H. Bull Chem. Soc.
Jpn. 1967, 40, 244-245.
[0103] 92. Furber, M.; Cooper, M. E.; Donald, D. K. Tetrahedron
Lett. 1993, 34, 1351-1354.
[0104] 93. Blair, Wade S.; Deshpande, Milind; Fang, Haiquan; Lin,
Pin-fang; Spicer, Timothy P.; Wallace, Owen B.; Wang, Hui; Wang,
Tao; Zhang, Zhongxing; Yeung, Kap-sun. Preparation of antiviral
indoleoxoacetyl piperazine derivatives U.S. Pat. No. 6,469,006.
Preparation of antiviral indoleoxoacetyl piperazine derivatives.
PCT Int. Appl. (PCT[US00/14359), WO 0076521 A1, filed May 24, 2000,
published Dec. 21, 2000.
[0105] 94. Wang, Tao; Wallace, Owen B.; Zhang, Zhongxing; Meanwell,
Nicholas A.; Bender, John A. Antiviral azaindole derivatives. U.S.
Pat. No. 6,476,034 and Wang, Tao; Wallace, Owen B.; Zhang,
Zhongxing; Meanwell, Nicholas A.; Bender, John A. Preparation of
antiviral azaindole derivatives. PCT Int. Appl. (PCT/US01/02009),
WO 0162255 A1, filed Jan. 19, 2001, published Aug. 30, 2001.
[0106] 95. Wallace, Owen B.; Wang, Tao; Yeung, Kap-Sun; Pearce,
Bradley C.; Meanwell, Nicholas A.; Qiu, Zhilei; Fang, Haiquan; Xue,
Qiufen May; Yin, Zhiwei. Composition and antiviral activity of
substituted indoleoxoacetic piperazine derivatives. U.S. patent
application Ser. No. 10/027,612 filed Dec. 19, 2001, which is a
continuation-in-part application of U.S. Ser. No. 09/888,686 filed
Jun. 25, 2001 (corresponding to PCT Int. Appl. (PCT/US01/20300), WO
0204440 A1, filed Jun. 26, 2001, published Jan. 17, 2002.
[0107] 96. J. L. Marco, S. T. Ingate, and P. M. Chinchon
Tetrahedron 1999, 55, 7625-7644.
[0108] 97. C. Thomas, F. Orecher, and P. Gmeiner Synthesis 1998,
1491.
[0109] 98. M. P. Pavia, S. J. Lobbestael, C. P. Taylor, F. M.
Hershenson, and D. W. Miskell.
[0110] 99. Buckheit, Robert W., Jr. Expert Opinion on
Investigational Drugs 2001, 10(8), 1423-1442.
[0111] 100. Balzarini, J.; De Clercq, E. Antiretroviral Therapy
2001, 31-62.
[0112] 101. E. De clercq Journal of Clinical Virology, 2001, 22,
73-89.
[0113] 102. Merour, Jean-Yves; Joseph, Benoit. Curr. Org. Chem.
(2001), 5(5), 471-506.
[0114] 103. T. W. von Geldern et al. J. Med. Chem 1996, 39,
968.
[0115] 104. M. Abdaoui et al. Tetrahedron 2000, 56, 2427. 105. W.
J. Spillane et al. J. Chem. Soc., Perkin Trans. 1, 1982, 3,
677.
[0116] 106. Wang, Tao; Wallace, Owen B.; Zhang, Zhongxing;
Meanwell, Nicholas A.; Kadow, John F. Yin, Zhiwei. Composition and
Antiviral Activity of Substituted Azaindoleoxoacetic piperazine
Derivatives. U.S. patent application Ser. No. 10/214,982 filed Aug.
7, 2002, which is a continuation-in-part application of U.S. Ser.
No. 10/038,306 filed Jan. 2, 2002 (corresponding to PCT Int. Appl.
(PCT/US02/00455), WO 02/062423 A1, filed Jan. 2, 2002, published
Aug. 15, 2002.
[0117] 107. Preparation of indolylglyoxylamides as antitumor
agents, Nickel, Bermd; Szelenyi, Istvan; Schmidt, Jurgen; Emig,
Peter; Reichert, Dietmar; Gunther, Eckhard; Brune, Kay, PCT Int.
Appl. WO 9951224, published Oct. 14, 1999.
SUMMARY OF THE INVENTION
[0118] The present invention comprises compounds of Formula I,
their pharmaceutical formulations, and their use in patients
suffering from or susceptible to a virus such as HIV. The compounds
of Formula I, which include nontoxic pharmaceutically acceptable
salts thereof, have the formula and meaning as described below.
[0119] The present invention comprises compounds of Formula I,
including pharmaceutically acceptable salts thereof, which are
effective antiviral agents, particularly as inhibitors of HIV.
[0120] An embodiment are compounds of Formula I, including
pharmaceutically acceptable salts thereof, 5
[0121] wherein:
[0122] Z is 6
[0123] Q is selected from the group consisting of 7
[0124] R.sup.1 is hydrogen;
[0125] R.sup.2, R.sup.3, R.sup.4, and R.sup.5, are independently
selected from the group consisting of hydrogen, halogen, cyano,
COOR.sup.8, XR.sup.9 and B;
[0126] m is 2;
[0127] R.sup.6 is O or does not exist;
[0128] R.sup.7 is hydrogen;
[0129] R.sup.10 is selected from the group consisting of
(C.sub.1-6)alkyl, --CH.sub.2CN, --CH.sub.2COOH,
--CH.sub.2C(O)NR.sup.11R.sup.12, phenyl and pyridinyl;
[0130] R.sup.11 and R.sup.12 are each independently H or
(C.sub.1-3)alkyl;
[0131] - - represents a carbon-carbon bond;
[0132] A is selected from the group consisting of cinnolinyl,
napthyridinyl, quinoxalinyl, pyridinyl, pyrimidinyl, quinolinyl,
isoquinolinyl, quinazolinyl, azabenzofuryl, and phthalazinyl each
of which may be optionally substituted with one or two groups
independently selected from methyl, methoxy, hydroxy, amino and
halogen;
[0133] --W-- is 8
[0134] R.sup.15, R.sup.16, R.sup.17, R.sup.18, R.sup.19, R.sup.20,
R.sup.21, R.sup.22 are each independently H or one of them is
methyl;
[0135] B is selected from the group consisting of
C(O)NR.sup.11R.sup.12C(.- dbd.NH)NHNHC(O)--R.sup.10,
C(.dbd.NH)cyclopropyl, C(.dbd.NOH)NH.sub.2, and heteroaryl; wherein
said heteroaryl is independently optionally substituted with a
substituent selected from F;
[0136] heteroaryl is selected from the group consisting of
pyridinyl, pyrazinyl, pyrimidinyl, thiazolyl, pyrrolyl, imidazolyl,
benzoimidazolyl, oxadiazolyl, pyrazolyl, tetrazolyl and
triazolyl;
[0137] F is selected from the group consisting of (C.sub.1-6)alkyl,
(C.sub.1-6)alkoxy, cyano, COOR.sup.8--CONR.sup.11R.sup.12;
--CH.sub.2CN, --CH.sub.2COOH, --CH.sub.2C(O)NR.sup.11R.sup.12,
phenyl and pyridinyl;
[0138] R.sup.8 and R.sup.9 are independently selected from the
group consisting of hydrogen and (C.sub.1-6)alkyl;
[0139] X is O;
[0140] provided that when A is pyridinyl or pyrimidinyl and Q is
9
[0141] then R.sup.5 is B.
[0142] Another embodiment are compounds of Formula I, including
pharmaceutically acceptable salts thereof,
[0143] wherein:
[0144] R.sup.15, R.sup.16, R.sup.17, R.sup.18, R.sup.19, R.sup.20,
R.sup.21, R.sup.22 are H;
[0145] R.sup.6 does not exist;
[0146] A is selected from members of the group consisting of 10
[0147] where Xw is the point of attachment and each member is
independently optionally substituted with one group selected from
the group consisting of methyl, methoxy, hydroxy, amino and
halogen;
[0148] Q is selected from the group consisting of 11
[0149] provided when Q is 12
[0150] then
[0151] R.sup.2 is hydrogen, methoxy or halogen; R.sup.3 and R.sup.4
are hydrogen; and R.sup.5 is selected from the group consisting of
hydrogen, halogen, cyano, COOR.sup.8, XR.sup.9 and B; or provided
when Q is 13
[0152] then
[0153] R.sup.2 is hydrogen, methoxy or halogen; R.sup.3 is
hydrogen; and R.sup.4 is selected from the group consisting of
hydrogen, halogen, cyano, COOR.sup.8, XR.sup.9 and B; or provided
when Q is 14
[0154] then
[0155] R.sup.2 and R.sup.3 are each hydrogen; and R.sup.4 is
selected from the group consisting of hydrogen, halogen, cyano,
COOR.sup.8, XR.sup.9 and B.
[0156] Another embodiment are compounds of Formula I, including
pharmaceutically acceptable salts thereof,
[0157] wherein:
[0158] B is selected from the group consisting of
C(O)NR.sup.11R.sup.12 and heteroaryl; wherein said heteroaryl is
independently optionally substituted with a substituent selected
from F;
[0159] heteroaryl is selected from the group consisting of
pyridinyl, pyrazinyl, pyrimidinyl, thiazolyl, pyrrolyl, imidazolyl,
benzoimidazolyl, oxadiazolyl, tetrazolyl and triazolyl.
[0160] Another embodiment are compounds of Formula I, including
pharmaceutically acceptable salts thereof,
[0161] wherein:
[0162] B is heteroaryl wherein said heteroaryl is independently
optionally substituted with a substituent selected from F.
[0163] Another embodiment are compounds of Formula I, including
pharmaceutically acceptable salts thereof,
[0164] wherein:
[0165] A is selected from the group consisting of 15
[0166] where Xw is the point of attachment.
[0167] Another embodiment are compounds of Formula I, including
pharmaceutically acceptable salts thereof,
[0168] wherein:
[0169] B is heteroaryl; wherein said heteroaryl is independently
optionally substituted with a substituent selected from F; and
[0170] heteroaryl is selected from the group consisting of
triazolyl, pyridinyl, pyrazinyl and pyrimidinyl.
[0171] Another embodiment are compounds of Formula I, including
pharmaceutically acceptable salts thereof,
[0172] wherein:
[0173] B is heteroaryl; wherein said heteroaryl is independently
optionally substituted with a substituent selected from F; and
[0174] heteroaryl is selected from the group consisting of
triazolyl.
[0175] Another embodiment are compounds of Formula I, including
pharmaceutically acceptable salts thereof,
[0176] wherein:
[0177] F is methyl.
[0178] Another embodiment is a pharmaceutical composition which
comprises an antiviral effective amount of a compound of Formula I,
including pharmaceutically acceptable salts thereof, as claimed in
claim 1, and one or more pharmaceutically acceptable carriers,
excipients or diluents; optionally which additionally comprises an
antiviral effective amount of an AIDS treatment agent selected from
the group consisting of:
[0179] (a) an AIDS antiviral agent;
[0180] (b) an anti-infective agent;
[0181] (c) an immunomodulator; and
[0182] (d) HIV entry inhibitors.
[0183] Another embodiment is a method for treating a mammal
infected with the HIV virus comprising administering to said mammal
an antiviral effective amount of a compound of Formula I, including
pharmaceutically accceptable salts thereof, and one or more
pharmaceutically acceptable carriers, excipients or diluents;
optionally in combination with an antiviral effective amount of an
AIDS treatment agent selected from the group consisting of an AIDS
antiviral agent; an anti-infective agent; an immunomodulator; and
an HIV entry inhibitor.
DETAILED DESCRIPTION OF THE INVENTION
[0184] Since the compounds of the present invention, may possess
asymmetric centers and therefore occur as mixtures of diastereomers
and enantiomers, the present invention includes the individual
diastereoisomeric and enantiomeric forms of the compounds of
Formula I in addition to the mixtures thereof.
Definitions
[0185] The term "C.sub.1-6 alkyl" as used herein and in the claims
(unless specified otherwise) mean straight or branched chain alkyl
groups such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl,
t-butyl, amyl, hexyl and the like.
[0186] "Halogen" refers to chlorine, bromine, iodine or
fluorine.
[0187] An "aryl" group refers to an all carbon monocyclic or
fused-ring polycyclic (i.e., rings which share adjacent pairs of
carbon atoms) groups having a completely conjugated pi-electron
system. Examples, without limitation, of aryl groups are phenyl,
napthalenyl and anthracenyl. The aryl group may be substituted or
unsubstituted. When substituted the substituted group(s) is
preferably one or more selected from alkyl, cycloalkyl, aryl,
heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy,
heteroaryloxy, heteroalicycloxy, thiohydroxy, thioaryloxy,
thioheteroaryloxy, thioheteroalicycloxy, cyano, halogen, nitro,
carbonyl, O-carbamyl, N-carbamyl, C-amido, N-amido, C-carboxy,
O-carboxy, sulfinyl, sulfonyl, sulfonamido, trihalomethyl, ureido,
amino and --NR.sup.xR.sup.y, wherein R.sup.x and R.sup.y are
independently selected from the group consisting of hydrogen,
alkyl, cycloalkyl, aryl, carbonyl, C-carboxy, sulfonyl,
trihalomethyl, and, combined, a five- or six-member heteroalicyclic
ring.
[0188] As used herein, a "heteroaryl" group refers to a monocyclic
or fused ring (i.e., rings which share an adjacent pair of atoms)
group having in the ring(s) one or more atoms selected from the
group consisting of nitrogen, oxygen and sulfur and, in addition,
having a completely conjugated pi-electron system. Unless otherwise
indicated, the heteroaryl group may be attached at either a carbon
or nitrogen atom within the heteroaryl group. It should be noted
that the term heteroaryl is intended to encompass an N-oxide of the
parent heteroaryl if such an N-oxide is chemically feasible as is
known in the art. Examples, without limitation, of heteroaryl
groups are furyl, thienyl, benzothienyl, thiazolyl, imidazolyl,
oxazolyl, oxadiazolyl, thiadiazolyl, benzothiazolyl, triazolyl,
tetrazolyl, isoxazolyl, isothiazolyl, pyrrolyl, pyranyl,
tetrahydropyranyl, pyrazolyl, pyridyl, pyrimidinyl, quinolinyl,
isoquinolinyl, purinyl, carbazolyl, benzoxazolyl, benzimidazolyl,
indolyl, isoindolyl, pyrazinyl. diazinyl, pyrazine,
triazinyltriazine, tetrazinyl, and tetrazolyl. When substituted the
substituted group(s) is preferably one or more selected from alkyl,
cycloalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy,
aryloxy, heteroaryloxy, heteroalicycloxy, thiohydroxy, thioaryloxy,
thioheteroaryloxy, thioheteroalicycloxy, cyano, halogen, nitro,
carbonyl, O-carbamyl, N-carbamyl, C-amido, N-amido, C-carboxy,
O-carboxy, sulfinyl, sulfonyl, sulfonamido, trihalomethyl, ureido,
amino, and --NR.sup.xR.sup.y, wherein R.sup.x and R.sup.y are as
defined above.
[0189] As used herein, a "heteroalicyclic" group refers to a
monocyclic or fused ring group having in the ring(s) one or more
atoms selected from the group consisting of nitrogen, oxygen and
sulfur. Rings are selected from those which provide stable
arrangements of bonds and are not intended to encomplish systems
which would not exist. The rings may also have one or more double
bonds. However, the rings do not have a completely conjugated
pi-electron system. Examples, without limitation, of
heteroalicyclic groups are azetidinyl, piperidyl, piperazinyl,
imidazolinyl, thiazolidinyl, 3-pyrrolidin-1-yl, morpholinyl,
thiomorpholinyl and tetrahydropyranyl. When substituted the
substituted group(s) is preferably one or more selected from alkyl,
cycloalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy,
aryloxy, heteroaryloxy, heteroalicycloxy, thiohydroxy, thioalkoxy,
thioaryloxy, thioheteroaryloxy, thioheteroalicycloxy, cyano,
halogen, nitro, carbonyl, thiocarbonyl, O-carbamyl, N-carbamyl,
O-thiocarbamyl, N-thiocarbamyl, C-amido, C-thioamido, N-amido,
C-carboxy, O-carboxy, sulfinyl, sulfonyl, sulfonamido,
trihalomethanesulfonamido, trihalomethanesulfonyl, silyl, guanyl,
guanidino, ureido, phosphonyl, amino and --NR.sup.xR.sup.y, wherein
R.sup.x and R.sup.y are as defined above.
[0190] An "alkyl" group refers to a saturated aliphatic hydrocarbon
including straight chain and branched chain groups. Preferably, the
alkyl group has 1 to 20 carbon atoms (whenever a numerical range;
e.g., "1-20", is stated herein, it means that the group, in this
case the alkyl group may contain 1 carbon atom, 2 carbon atoms, 3
carbon atoms, etc. up to and including 20 carbon atoms). More
preferably, it is a medium size alkyl having 1 to 10 carbon atoms.
Most preferably, it is a lower alkyl having 1 to 4 carbon atoms.
The alkyl group may be substituted or unsubstituted. When
substituted, the substituent group(s) is preferably one or more
individually selected from trihaloalkyl, cycloalkyl, aryl,
heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy,
heteroaryloxy, heteroalicycloxy, thiohydroxy, thioalkoxy,
thioaryloxy, thioheteroaryloxy, thioheteroalicycloxy, cyano, halo,
nitro, carbonyl, thiocarbonyl, O-carbamyl, N-carbamyl,
O-thiocarbamyl, N-thiocarbamyl, C-amido, C-thioamido, N-amido,
C-carboxy, O-carboxy, sulfinyl, sulfonyl, sulfonamido,
trihalomethanesulfonamido, trihalomethanesulfonyl, and combined, a
five- or six-member heteroalicyclic ring.
[0191] A "cycloalkyl" group refers to an all-carbon monocyclic or
fused ring (i.e., rings which share and adjacent pair of carbon
atoms) group wherein one or more rings does not have a completely
conjugated pi-electron system. Examples, without limitation, of
cycloalkyl groups are cyclopropane, cyclobutane, cyclopentane,
cyclopentene, cyclohexane, cyclohexadiene, cycloheptane,
cycloheptatriene and adamantane. A cycloalkyl group may be
substituted or unsubstituted. When substituted, the substituent
group(s) is preferably one or more individually selected from
alkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy,
heteroaryloxy, heteroalicycloxy, thiohydroxy, thioalkoxy,
thioaryloxy, thioheteroaryloxy, thioheteroalicycloxy, cyano, halo,
nitro, carbonyl, thiocarbonyl, O-carbamyl, N-carbamyl,
O-thiocarbamyl, N-thiocarbamyl, C-amido, C-thioamido, N-amido,
C-carboxy, O-carboxy, sulfinyl, sulfonyl, sulfonamido,
trihalo-methanesulfonamido, trihalomethanesulfonyl, silyl, guanyl,
guanidino, ureido, phosphonyl, amino and --NR.sup.xR.sup.y with
R.sup.x and R.sup.y as defined above.
[0192] An "alkenyl" group refers to an alkyl group, as defined
herein, consisting of at least two carbon atoms and at least one
carbon-carbon double bond.
[0193] An "alkynyl" group refers to an alkyl group, as defined
herein, consisting of at least two carbon atoms and at least one
carbon-carbon triple bond.
[0194] A "hydroxy" group refers to an --OH group.
[0195] An "alkoxy" group refers to both an --O-alkyl and an
--O-cycloalkyl group as defined herein.
[0196] An "aryloxy" group refers to both an --O-aryl and an
--O-heteroaryl group, as defined herein.
[0197] A "heteroaryloxy" group refers to a heteroaryl-O-- group
with heteroaryl as defined herein.
[0198] A "heteroalicycloxy" group refers to a heteroalicyclic-O--
group with heteroalicyclic as defined herein.
[0199] A "thiohydroxy" group refers to an --SH group.
[0200] A "thioalkoxy" group refers to both an S-alkyl and an
--S-cycloalkyl group, as defined herein.
[0201] A "thioaryloxy" group refers to both an --S-aryl and an
--S-heteroaryl group, as defined herein.
[0202] A "thioheteroaryloxy" group refers to a heteroaryl-S-- group
with heteroaryl as defined herein.
[0203] A "thioheteroalicycloxy" group refers to a
heteroalicyclic-S-- group with heteroalicyclic as defined
herein.
[0204] A "carbonyl" group refers to a --C(.dbd.O)--R" group, where
R" is selected from the group consisting of hydrogen, alkyl,
alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl (bonded through a
ring carbon) and heteroalicyclic (bonded through a ring carbon), as
each is defined herein.
[0205] An "aldehyde" group refers to a carbonyl group where R" is
hydrogen.
[0206] A "thiocarbonyl" group refers to a --C(.dbd.S)--R" group,
with R" as defined herein.
[0207] A "Keto" group refers to a --CC(.dbd.O)C-- group wherein the
carbon on either or both sides of the C.dbd.O may be alkyl,
cycloalkyl, aryl or a carbon of a heteroaryl or heteroaliacyclic
group.
[0208] A "trihalomethanecarbonyl" group refers to a
Z.sub.3CC(.dbd.O)-- group with said Z being a halogen.
[0209] A "C-carboxy" group refers to a --C(.dbd.O)O--R" groups,
with R" as defined herein.
[0210] An "O-carboxy" group refers to a R"C(--O)O-group, with R" as
defined herein.
[0211] A "carboxylic acid" group refers to a C-carboxy group in
which R" is hydrogen.
[0212] A "trihalomethyl" group refers to a --CZ.sub.3, group
wherein Z is a halogen group as defined herein.
[0213] A "trihalomethanesulfonyl" group refers to an
Z.sub.3CS(.dbd.O).sub.2-- groups with Z as defined above.
[0214] A "trihalomethanesulfonamido" group refers to a
Z.sub.3CS(.dbd.O).sub.2NR.sup.x-- group with Z and Rx as defined
herein.
[0215] A "sulfinyl" group refers to a --S(.dbd.O)--R" group, with
R" as defined herein and, in addition, as a bond only; i.e.,
--S(O)--.
[0216] A "sulfonyl" group refers to a --S(.dbd.O).sub.2R" group
with R" as defined herein and, in addition as a bond only; i.e.,
--S(O).sub.2--.
[0217] A "S-sulfonamido" group refers to a
--S(.dbd.O).sub.2NR.sup.XR.sup.- Y, with R.sup.X and R.sup.Y as
defined herein.
[0218] A "N-Sulfonamido" group refers to a
R"S(.dbd.O).sub.2NR.sub.X-- group with R.sub.x as defined
herein.
[0219] A "O-carbamyl" group refers to a --OC(.dbd.O)NR.sup.xR.sup.y
as defined herein.
[0220] A "N-carbamyl" group refers to a R.sup.xOC(.dbd.O)NR.sup.y
group, with R.sup.x and R.sup.y as defined herein.
[0221] A "O-thiocarbamyl" group refers to a
--OC(.dbd.S)NR.sup.xR.sup.y group with R.sup.x and R.sup.y as
defined herein.
[0222] A "N-thiocarbamyl" group refers to a
R.sup.xOC(.dbd.S)NR.sup.y-- group with R.sup.x and R.sup.y as
defined herein.
[0223] An "amino" group refers to an --NH.sub.2 group.
[0224] A "C-amido" group refers to a --C(.dbd.O)NR.sup.xR.sup.y
group with R.sup.x and R.sup.y as defined herein.
[0225] A "C-thioamido" group refers to a --C(.dbd.S)NR.sup.xR.sup.y
group, with R.sup.x and R.sup.y as defined herein.
[0226] A "N-amido" group refers to a R.sup.xC(.dbd.O)NR.sup.y--
group, with R.sup.x and R.sup.y as defined herein.
[0227] An "ureido" group refers to a
--NR.sup.xC(.dbd.O)NR.sup.yR.sup.y2 group with R.sup.x and R.sup.y
as defined herein and R.sup.y2 defined the same as R.sup.x and
R.sup.y.
[0228] An "thioureido" group refers to a
--NR.sup.xC(.dbd.S)NR.sup.yR.sup.- y2 group with R.sup.x and
R.sup.y as defined herein and R.sup.y2 defined the same as R.sup.x
and R.sup.y.
[0229] A "guanidino" group refers to a
--R.sup.xNC(.dbd.N)NR.sup.yR.sup.y2 group, with R.sup.x, R.sup.y
and R.sup.y2 as defined herein.
[0230] A "guanyl" group refers to a R.sup.xR.sup.yNC(.dbd.N)--
group, with R.sup.x and R.sup.y as defined herein.
[0231] A "cyano" group refers to a --CN group.
[0232] A "silyl" group refers to a --Si(R").sub.3, with R" as
defined herein.
[0233] A "phosphonyl" group refers to a P(.dbd.O)(OR.sup.x).sub.2
with R.sup.x as defined herein.
[0234] A "hydrazino" group refers to a --NR.sup.xNR.sup.yR.sup.y2
group with R.sup.x, R.sup.y and R.sup.y2 as defined herein.
[0235] Any two adjacent R groups may combine to form an additional
aryl, cycloalkyl, heteroaryl or heterocyclic ring fused to the ring
initially bearing those R groups.
[0236] It is known in the art that nitogen atoms in heteroaryl
systems can be "participating in a heteroaryl ring double bond",
and this refers to the form of double bonds in the two tautomeric
structures which comprise five-member ring heteroaryl groups. This
dictates whether nitrogens can be substituted as well understood by
chemists in the art. The disclosure and claims of the present
invention are based on the known general principles of chemical
bonding. It is understood that the claims do not encompass
structures known to be unstable or not able to exist based on the
literature.
[0237] Physiologically acceptable salts and prodrugs of compounds
disclosed herein are within the scope of this invention. 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. The term "pharmaceutically acceptable salt" as used
herein is also intended to include salts of acidic groups, such as
a carboxylate, with such counterions as ammonium, alkali metal
salts, particularly sodium or potassium, alkaline earth metal
salts, particularly calcium or magnesium, and salts with suitable
organic bases such as lower alkylamines (methylamine, ethylamine,
cyclohexylamine, and the like) or with substituted lower
alkylamines (e.g. hydroxyl-substituted alkylamines such as
diethanolamine, triethanolamine or
tris(hydroxymethyl)-aminomethane), or with bases such as piperidine
or morpholine.
[0238] 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 the 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
therapeutic 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 HIV infection.
[0239] 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, immunomodulators, antiinfectives, or vaccines, such as
those in the following table.
[0240] The invention also encompasses methods where the compound is
given in combination therapy. That is, the compound can be used in
conjunction with, but separately from, other agents useful in
treating AIDS and HIV infection. Some of these agents include HIV
attachment inhibitors, CCR5 inhibitors, CXCR4 inhibitors, HIV cell
fusion inhibitors, HIV integrase inhibitors, HIV nucleoside reverse
transcriptase inhibitors, HIV non-nucleoside reverse transcriptase
inhibitors, HIV protease inhibitors, budding and maturation
inhibitors, immunomodulators, and anti-infectives. In these
combination methods, the compounds of this invention will generally
be given in a daily dose of 1-100 mg/kg body weight daily in
conjunction with other agents. The other agents generally will be
given in the amounts used therapeutically. The specific dosing
regime, however, will be determined by a physician using sound
medical judgement.
[0241] Table 2 lists some agents useful in treating AIDS and HIV
infection which are suitable for this invention.
1TABLE 2 Drug Name Manufacturer Indication Antivirals 097
Hoechst/Bayer HIV infection, AIDS, (non-nucleoside ARC reverse
transcriptase inhibitor) Amprenavir Glaxo Wellcome HIV infection,
AIDS, 141 W94 ARC GW 141 (protease inhibitor) Abacavir (1592U89)
Glaxo Wellcome HIV infection, AIDS, GW 1592 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, ARC, AL-721 Ethigen PGL HIV positive, (Los Angeles, CA)
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 AIDS,
ARC Neutralizes pH Biotherapy Labile alpha Concepts aberrant
(Rockville, MD) Interferon AR177 Aronex Pharm HIV infection, AIDS,
ARC Beta-fluoro-ddA Nat'l Cancer AIDS-associated Institute diseases
BMS-232623 Bristol-Myers HIV infection, AIDS, (CGP-73547)
Squibb/Novartis ARC (protease inhibitor) BMS-234475 Bristol-Myers
HIV infection, AIDS, (CGP-61755) Squibb/Novartis 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, (RT inhibitor) ARC Dextran Sulfate Ueno Fine Chem.
AIDS, ARC, HIV Ind. Ltd. (Osaka, positive Japan) asymptomatic ddC
Hoffman-La Roche HIV infection, AIDS, Dideoxycytidine ARC ddI
Bristol-Myers HIV infection, AIDS, Dideoxyinosine Squibb ARC;
combination with AZT/d4T DMP-450 AVID HIV infection, AIDS,
(protease inhibitor) (Camden, NJ) ARC Efavirenz DuPont Merck HIV
infection, AIDS, (DMP 266) ARC (-)6-Chloro-4-(S)-
cyclopropylethynyl- 4(S)-trifluoro- methyl-1,4-dihydro-
2H-3,1-benzoxazin- 2-one, STOCRINE (non-nucleoside RT inhibitor)
EL10 Elan Corp, PLC HIV infection (Gainesville, GA) Famciclovir
Smith Kline herpes zoster, herpes simplex FTC(reverse Emory
University HIV infection, AIDS, transcriptase ARC inhibitor) GS 840
Gilead HIV infection, AIDS, (reverse ARC transcriptase inhibitor)
HBY097 Hoechst Marion HIV infection, AIDS, (non-nucleoside Roussel
ARC 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 ARC, AIDS Sciences 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 HIV-associated Institute diseases
Lamivudine, 3TC Glaxo Wellcome HIV infection, AIDS, (reverse ARC,
also with AZT transcriptase inhibitor) Lobucavir Bristol-Myers CMV
infection Squibb Nelfinavir Agouron HIV infection, AIDS, (protease
Pharmaceuticals ARC inhibitor) Nevirapine Boeheringer HIV
infection, AIDS, (RT inhibitor) Ingleheim ARC Novapren Novaferon
Labs, HIV inhibitor Inc. (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-140690 Pharmacia Upjohn HIV infection, AIDS,
(protease inhibitor) ARC Probucol Vyrex HIV infection, AIDS RBC-CD4
Sheffield Med. HIV infection, AIDS, Tech (Houston, TX) ARC
Ritonavir Abbott HIV infection, AIDS, (protease inhibitor) ARC
Saquinavir Hoffmann- HIV infection, AIDS, (protease inhibitor)
LaRoche ARC Stavudine; d4T Bristol-Myers HIV infection, AIDS,
Didehydrodeoxy- Squibb ARC thymidine Valaciclovir Glaxo Wellcome
Genital HSV & CMVinfections 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 Tenofovir Gilead HIV infection, AIDS disoproxil, fumarate
salt (Viread .RTM.) (reverse tran- scriptase inhibitor) Combivir
.RTM. GSK HIV infection, AIDS (reverse transcriptase inhibitor)
abacavir succinate GSK HIV infection, AIDS (or Ziagen .RTM.)
(reverse transcriptase inhibitor) Reyataz .RTM. Bristol-Myers HIV
infection, AIDS (atazanavir) Squibb Fuzeon Roche/Trimeris HIV
infection, AIDS, (Enfuvirtide, viral fusion T-20) inhibitor
Trizivir .RTM. HIV infection, AIDS Kaletra .RTM. Abbott HIV
infection, AIDS, ARC Immunomodulators AS-101 Wyeth-Ayerst AIDS
Bropirimine Pharmacia Upjohn Advanced AIDS Acemannan Carrington
Labs, AIDS, ARC Inc. (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, combination Macrophage Colony
w/AZT Stimulating Factor HIV Core Particle Rorer Seropositive HIV
Immunostimulant IL-2 Cetus AIDS, in combination Interleukin-2 w/AZT
IL-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 Amgen AIDS, in combination Granulocyte w/AZT
Colony Stimulating 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-La Roche Kaposi's sarcoma, Alfa 2a in
combination AIDS, ARC 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
Clindamycin with Pharmacia Upjohn PCP Primaquine Fluconazole Pfizer
Cryptococcal meningitis, candidiasis Pastille Squibb Corp.
Prevention of oral Nystatin Pastille candidiasis Ornidyl Merrell
Dow PCP Eflornithine Pentamidine LyphoMed PCP treatment Isethionate
(IM & IV) (Rosemont, IL) Trimethoprim Antibacterial
Trimethoprim/ Antibacterial sulfa 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 assoc. Erythropoietin with AZT
therapy Recombinant Human Serono AIDS-related wasting, Growth
Hormone cachexia Megestrol Acetate Bristol-Myers Treatment of
Squibb anorexia assoc. W/AIDS Testosterone Alza, Smith Kline
AIDS-related wasting Total Enteral Norwich Eaton Diarrhea and
Nutrition Pharmaceuticals malabsorption related to AIDS
[0242] 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, and
Meanwell, Nicholas A.; Kadow, John F. Inhibitors of the entry of
HIV into host cells. Current Opinion in Drug Discovery &
Development (2003), 6(4), 451-461.
[0243] 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.
[0244] 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 or the compound may be combined with one or
two nucleoside inhibitor of HIV reverse transcriptase. An optional
fourth component in the combination from the list of available
drugs may be added. A preferred inhibitor of HIV protease is
Reyataz.RTM. (atazanavir sulfate). Reyataz.RTM. is generally
administered at a dosage of 400 mg once a day but may also be
administered in combination with Ritonavir.RTM.. Another preferred
protease inhibitor is Kaletra.RTM.. Preferred non-nucleoside
inhibitors of HIV reverse transcriptase include efavirenz. The
compounds in this invention could be administered in combination
with Emtriva.RTM. (emtricitabine) and Viread.RTM. (Tenofovir
dipivoxil) for example. These compounds are typically administered
at doses of 200 mg or 300 mg once daily respectively. These
combinations may have unexpected effects on limiting the spread and
degree of infection of HIV.
[0245] 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).
Abbreviations
[0246] The following abbreviations, most of which are conventional
abbreviations well known to those skilled in the art, are used
throughout the description of the invention and the examples. Some
of the abbreviations used are as follows:
[0247] h=hour(s)
[0248] rt=room temperature
[0249] mol=mole(s)
[0250] mmol=millimole(s)
[0251] g=gram(s)
[0252] mg=milligram(s)
[0253] mL=milliliter(s)
[0254] TFA=Trifluoroacetic Acid
[0255] DCE=1,2-Dichloroethane
[0256] CH.sub.2Cl.sub.2=Dichloromethane
[0257] TPAP=tetrapropylammonium perruthenate
[0258] THF=Tetrahydofuran
[0259]
DEPBT=3-(Diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one
[0260] DMAP=4-dimethylaminopyridine
[0261] P-EDC=Polymer supported
1-(3-dimethylaminopropyl)-3-ethylcarbodiimi- de
[0262] EDC=1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
[0263] DMF=N,N-dimethylformamide
[0264] Hunig's Base .dbd.N,N-Diisopropylethylamine
[0265] mCPBA=meta-Chloroperbenzoic Acid
[0266] azaindole=1H-Pyrrolo-pyridine
[0267] 4-azaindole=1H-pyrrolo[3,2-b]pyridine
[0268] 5-azaindole=1H-Pyrrolo[3,2-c]pyridine
[0269] 6-azaindole=1H-pyrrolo[2,3-c]pyridine
[0270] 7-azaindole=1H-Pyrrolo[2,3-b]pyridine
[0271] PMB=4-Methoxybenzyl
[0272] DDQ=2,3-Dichloro-5,6-dicyano-1,4-benzoquinone
[0273] OTf=Trifluoromethanesulfonoxy
[0274] NMM=4-Methylmorpholine
[0275] PIP--COPh=1-Benzoylpiperazine
[0276] NaHMDS=Sodium hexamethyldisilazide
[0277] EDAC=1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide
[0278] TMS=Trimethylsilyl
[0279] DCM=Dichloromethane
[0280] DCE=Dichloroethane
[0281] MeOH=Methanol
[0282] THF=Tetrahydrofuran
[0283] EtOAc=Ethyl Acetate
[0284] LDA=Lithium diisopropylamide
[0285] TMP--Li=2,2,6,6-tetramethylpiperidinyl lithium
[0286] DME=Dimethoxyethane
[0287] DIBALH=Diisobutylaluminum hydride
[0288] HOBT=1-hydroxybenzotriazole
[0289] CBZ=Benzyloxycarbonyl
[0290] PCC=Pyridinium chlorochromate
[0291] Chemistry
[0292] The present invention comprises compounds of Formula I,
their pharmaceutical formulations, and their use in patients
suffering from or susceptible to HfV infection. The compounds of
Formula I include pharmaceutically acceptable salts thereof.
[0293] The synthesis procedures and anti-HIV-1 activities of
substituted indole or azaindole oxoacetic N-heteroaryl piperazine
containing analogs are described below. Scheme A depicts a typical
method of completing the synthesis of the compounds of claim 1.
Coupling of the appropriate oxo acetic acid with the desired N-aryl
piperazine or its acid salt can be carried out using a variety of
conditions as described for step D. 16
[0294] Step D. The acid intermediate Z-OH from Scheme A (which can
also be depicted as intermediates QC(O)C(O)OH) or 4a-e, from step C
of Schemes 1a-1e respectively are coupled with either a substituted
piperazine, H--W-A as shown in Schemes A and 1a-1e or a protected
piperazine, for example t-butyl 1-piperazinecarboxylate
(Boc-piperazine, H--W-tBoc), as shown in Scheme (where W
corresponds to the W in Formula I and H is hydrogen). They can be
coupled with the acid using standard amide bond or peptide bond
forming coupling reagents. The combination of EDAC and
triethylamine in tetrahydrofuran or BOPCl and diisopropyl ethyl
amine in chloroform have been utilized most frequently but DEPBT,
or other coupling reagents such as PyBop could be utilized. Another
useful coupling condition employs HATU (L. A. Carpino et. al. J.
Chem. Soc. Chem Comm. 1994, 201-203; A. Virgilio et. al. J. Am.
Chem. Soc. 1994, 116,11580-11581). A general procedure for using
this reagent is Acid (1eq) and H--W-Boc or H--W--SO.sub.2-A or HCl
salt (2eq) in DMF are stirred at rt for between 1 h and 2 days.
HATU (2eq) was added in one portion and then DMAP (3eq). The
reaction was stirred at rt for 2 to 15 h (reaction progress
monitored by standard methods ie TLC, LC/MS). The mixture is
filtered through filter paper to collect the solid. The filtrate is
concentrated and water is added. The mixture is filtered again and
the solid is washed with water. The solid is conbined and washed
with water. Many reagents for amide bond couplings are known by an
organic chemist skilled in the art and nearly all of these are
applicable for realizing coupled amide products.
[0295] As mentioned above, DEPBT
(3-(diethoxyphosphoryloxy)-1,2,3-benzotri- azin-4(3H)-one) and
N,N-diisopropylethylamine, commonly known as Hunig's base,
represents another efficient method to form the amide bond (step D)
and provide compounds of claim 1. DEPBT is either purchased from
Adrich or prepared according to the procedure of Ref. 28, Li, H.;
Jiang, X.; Ye, Y.-H.; Fan, C.; Romoff, T.; Goodman, M. Organic
Lett., 1999, 1, 91-93. Typically an inert solvent such as DMF or
THF is used but other aprotic solvents could be used.
[0296] The amide bond construction reaction could be carried out
using the preferred conditions described above, the EDC conditions
described below, other coupling conditions described in this
application, or alternatively by applying the conditions or
coupling reagents for amide bond construction described later in
this application for construction of substituents R.sub.2-R.sub.5.
Some specific nonlimiting examples are given in this
application.
[0297] Alternatively, the acid could be converted to a methyl ester
using excess diazomethane in THF/ether. The methyl ester in dry THF
could be reacted with the lithium amide of intermediate H--W. The
lithium amide of H--W, Li--W is formed by reacting intermediate 1
with lithium bistrimethylsilylamide in THF for 30 minutes in an ice
water cooling bath. Sodium or potassium amides could be formed
similarly and utilized if additional reactivity is desired. Other
esters such as ethyl, phenyl, or pentafluorophenyl could be
utilized and would be formed using standard methodology.
[0298] The amide bond construction reaction could be carried out
using the preferred conditions described above, the EDC conditions
described below, other coupling conditions described in this
application, or alternatively by applying the conditions or
coupling reagents for amide bond construction described later in
this application for construction of substituents R.sub.2-R.sub.5.
Some specific nonlimiting examples are given in this application.
In addition, the acid can be converted to the acid chloride using
oxalyl chloride in a solvent such as benzene or thionyl chloride
either neat or containing a catalystic amount of DMF. Temperatures
between 0.degree. C. and reflux may be utilized depending on the
substrate. Compounds of Formula I can be obtained from the
resultant compounds of formula Z-Cl by reaction with the
appropriate H--W-A in the presence of a tertiary amine (3-10 eq.)
such as triethylamine or diisopropylethylamine in an anhydrous
aprotic solvent such as dichloromethane, dichloroethane, diethyl
ether, dioxane, THF, acetonitrile, DMF or the like at temperatures
ranging from 0.degree. C. to reflux. Most preferred are
dichloromethane, dichloroethane, or THF. The reaction can be
monitored by LC/MS.
[0299] It should be noted that in many cases reactions are depicted
for only one position of an intermediate, such as the R.sup.5
position, for example. It is to be understood that such reactions
could be used at other positions, such as R.sup.2-R.sup.4, of the
various intermediates. Reaction conditions and methods given in the
specific examples are broadly applicable to compounds with other
substitution and other tranformations in this application. Schemes
A and 1a-1e describe general reaction schemes for taking
appropriately substituted Q (indoles and azaindoles) and converting
them to compounds of Formula I. While these schemes are very
general, other permutations such as carrying a precursor or
precursors to substituents R.sup.2 through R.sup.5 through the
reaction scheme and then converting it to a compound of Formula I
in the last step are also contemplated methods of this invention.
Nonlimiting examples of such strategies follow in subsequent
schemes. Procedures for coupling piperazine amides to oxoacetyl
derivatives are described in the Blair, Wang, Wallace, or Wang
references 93-95 and 106 respectively. The entire disclosures in
U.S. Pat. No. 6,469,006 granted Oct. 22, 2002; U.S. Pat. No.
6,476,034 granted Nov. 5, 2002; U.S. patent application Ser. No.
10/027,612 filed Dec. 19, 2001, which is a continuation-in-part of
U.S. Ser. No. 09/888,686 filed Jun. 25, 2001 (corresponding to PCT
WO 02/04440, published Jan. 17, 2002); and U.S. patent application
Ser. No. 10/214,982 filed Aug. 7, 2002, which is a
continuation-in-part of U.S. Ser. No. 10/038,306 filed Jan. 2, 2002
(corresponding to PCT WO 02/62423 published Aug. 15, 2002) are
incorporated by reference herein. The procedures used to couple
indole or azaindole oxoacetic acids to piperazine amides in these
references can be used analogously to form the compounds of this
invention except the N-heteroaryl piperazines are used in place of
the piperazine benzamides. It should be stated that the procedures
incorporated from these applications encompass the preparation of
starting materials and transformations which are useful for
enabling the preparation of compounds of this invention.
[0300] Procedures for making Z (as defined in Formula I of the
description of the invention) are described in the Blair, Wang,
Wallace, or Wang references 93-95 and 106 respectively. The entire
disclosures in U.S. Pat. No. 6,469,006 granted Oct. 22, 2002; U.S.
Pat. No. 6,476,034 granted Nov. 5, 2002; U.S. patent application
Ser. No. 10/027,612 filed Dec. 19, 2001, which is a
continuation-in-part of U.S. Ser. No. 09/888,686 filed Jun. 25,
2001 (corresponding to PCT WO 02/04440, published Jan. 17, 2002);
and U.S. patent application Ser. No. 10/214,982 filed Aug. 7, 2002,
which is a continuation-in-part of U.S. Ser. No. 10/038,306 filed
Jan. 2, 2002 (corresponding to PCT WO 02/62423 published Aug. 15,
2002) are incorporated by reference herein.
[0301] Additional general procedures to construct substituted
azaindole Q and Z of Formula I and intermediates useful for their
synthesis are described in the following Schemes. 17 18 19 20
21
[0302] Step A. In Schemes 1a-1e depict the synthesis of a aza
indole or indole intermediates, 2a-2e via the well known Bartoli
reaction in which vinyl magnesium bromide reacts with an aryl or
heteroaryl nitro group, such as in 1a-1e, to form a five-membered
nitrogen containing ring as shown. Some references for details on
how to carry out the transformation include: Bartoli et al. a)
Tetrahedron Lett. 1989, 30, 2129. b) J. Chem. Soc. Perkin Trans. 1
1991, 2757. c) J. Chem. Soc. Perkin Trans. II 1991, 657. d)
Synthesis (1999), 1594. e) Zhang, Zhongxing; Yang, Zhong; Meanwell,
Nicholas A.; Kadow, John F.; Wang, Tao. "A General Method for the
Preparation of 4- and 6-Azaindoles". Journal of Organic Chemistry
2002, 67 (7), 2345-2347 WO 02/62423 Aug. 15, 2002 "Preparation and
antiviral activity for HIV-I of substituted
azaindoleoxoacetylpiperazines- " Wang, Tao; Zhang, Zhongxing;
Meanwell, Nicholas A.; Kadow, John F.; Yin, Zhiwei.
[0303] In the preferred procedure, a solution of vinyl Magnesium
bromide in THF (typically 1.0M but from 0.25 to 3.0M) is added
dropwise to a solution of the nitro pyridine in THF at -78.degree.
under an inert atmosphere of either nitrogen or Argon. After
addition is completed, the reaction temperature is allowed to warm
to -20.degree. and then is stirred for approximately 12 h before
quenching with 20% aq ammonium chloride solution. The reaction is
extracted with ethyl acetate and then worked up in a typical manner
using a drying agent such as anhydrous magnesium sulfate or sodium
sulfate. Products are generally purified using chromatography over
Silica gel. Best results are generally achieved using freshly
prepared vinyl Magnesium bromide. In some cases, vinyl Magnesium
chloride may be substituted for vinyl Magnesium bromide. In some
cases modified procedures might occasionally provide enhanced
yield. An inverse addition procedure can sometimes be employed.
(The nitro pyridine solution is added to the vinyl Grignard
solution). Occasionally solvents such as dimethoxy ethane or
dioxane may prove useful. A procedure in which the nitro compound
in THF is added to a 1M solution of vinyl magnesium bromide in THF
at -40.degree. C. may prove beneficial. Following completion of the
reaction by TLC the reaction is quenched with sat ammonium chloride
aqueous solution and purified by standard methods. A reference for
this alternative procedure is contained in M. C. Pirrung, M. Wedel,
and Y. Zhao et. al. Syn Lett 2002, 143-145.
[0304] Substituted azaindoles may be prepared by methods described
in the literature or may be available from commercial sources. Thus
there are many methods for synthesizing intermediates 2a-2d and the
specific examples are too numerous to even list. Methodology for
the preparation of many compounds of interest is described in
references of Blair, Wang, Wallace, and Wang references 93-95 and
106 respectively. A review on the synthesis of 7-azaindoles has
been published (Merour et. al. reference 102). Alternative
syntheses of aza indoles and general methods for synthesizing
intermediates 2 include, but are not limited to, those described in
the following references (a-k below): a) Prokopov, A. A.;
Yakhontov, L. N. Khim.-Farm. Zh. 1994, 28(7), 30-51; b)
Lablache-Combier, A. Heteroaromatics. Photoinduced Electron
Transfer 1988, Pt. C, 134-312; c) Saify, Zafar Said. Pak. J.
Pharmacol. 1986, 2(2), 43-6; d) Bisagni, E. Jerusalem Symp. Quantum
Chem. Biochem. 1972, 4, 439-45; e) Yakhontov, L. N. Usp. Khim.
1968, 37(7), 1258-87; f) Willette, R. E. Advan. Heterocycl. Chem.
1968, 9, 27-105; g) Mahadevan, I.; Rasmussen, M. Tetrahedron 1993,
49(33), 7337-52; h) Mahadevan, I.; Rasmussen, M. J. Heterocycl.
Chem. 1992, 29(2), 359-67; i) Spivey, A. C.; Fekner, T.; Spey, S.
E.; Adams, H. J. Org. Chem. 1999, 64(26), 9430-9443; j) Spivey, A.
C.; Fekner, T.; Adams, H. Tetrahedron Lett. 1998, 39(48),
8919-8922; k) Advances in Heterocyclic Chemistry (Academic press)
1991, Vol. 52, pg 235-236 and references therein. Other references
later in this application. Starting indole intermediates of formula
2e (Scheme 10) are known or are readily prepared according to
literature procedures, such as those described in Gribble, G.
(Refs. 24 and 99), Bartoli et al (Ref. 36), reference 37, or the
book by Richard A. Sundberg in reference 40. Other methods for the
preparation of indole intermediates include: the Leimgruber-Batcho
Indole synthesis (reference 93); the Fisher Indole synthesis
(references 94 and 95); the 2,3-rearrangement protocol developed by
Gassman (reference 96); the annelation of pyrroles (reference 97);
tin mediated cyclizations (reference 98); and the Larock palladium
mediated cyclization of 2-alkynyl anilines. Many other methods of
indole synthesis are known and a chemist with typical skill in the
art can readily locate conditions for preparation of indoles which
can be utilized to prepare compounds of Formula I. 22
[0305] Scheme 1f depicts a shorthand method for representing the
intermediates used for reactions in Schemes 1a-1c, and Schemes 2-7
and generic Q. It is understood, for the purposes of Scheme 1f and
further Schemes, that 1b is used to synthesize 2b-5b, 1c provides
2c-5c and 1d provides 2d-5d etc. The substituents R.sub.x represent
for azaindoles R.sub.2-R.sub.4 and for indoles R.sub.2-R.sub.5. In
formulas in following schemes, one of the substituents may be
depicted but it is understood that each formula can represent the
appropriate generic azaindoles or indole in order to keep the
application succinct.
[0306] Step B. Intermediates 3a-e can be prepared by reaction of
indoles or azaindoles (intermediates 2), with an excess of
ClCOCOOMe in the presence of AlCl.sub.3 (aluminum chloride)
(Sycheva et al, Ref. 26, Sycheva, T. V.; Rubtsov, N. M.; Sheinker,
Yu. N.; Yakhontov, L. N. Some further descriptions of the exact
procedures to carry out this reaction are contained in a) Zhang,
Zhongxing; Yang, Zhong; Wong, Henry; Zhu, Juliang; Meanwell,
Nicholas A.; Kadow, John F.; Wang, Tao. "An Effective Procedure for
the Acylation of Azaindoles at C-3." J. Org. Chem. 2002, 67(17),
6226-6227; b) Tao Wang et. al. U.S. Pat. No. 6,476,034 B2
"Antiviral Azaindole derivatives" published Nov. 5, 2002; c) W.
Blair et al. PCT patent application WO 00/76521 A1 published Dec.
21, 2000; d) 0. Wallace et. al. PCT application WO 02/04440A1
published Jan. 17, 2002. Some reactions of
5-cyano-6-chloro-7-azaindoles and lactam-lactim tautomerism in
5-cyano-6-hydroxy-7-azaindolines. Khim. Geterotsikl. Soedin., 1987,
100-106). Typically an inert solvent such as CH.sub.2Cl.sub.2 is
used but others such as THF, Et.sub.2O, DCE, dioxane, benzene, or
toluene may find applicability either alone or in mixtures. Other
oxalate esters such as ethyl or benzyl mono esters of oxalic acid
could also suffice for either method shown above. More lipophilic
esters ease isolation during aqueous extractions. Phenolic or
substituted phenolic (such as pentafluorophenol) esters enable
direct coupling of the HW-protecting group, such as a
Boc-piperazine, in Step D without activation. Lewis acid catalysts,
such as tin tetrachloride, titanium IV chloride, and aluminum
chloride are employed in Step B with aluminum chloride being most
preferred. Alternatively, the azaindole is treated with a Grignard
reagent such as MeMgI (methyl magnesium iodide), methyl magnesium
bromide or ethyl magnesium bromide and a zinc halide, such as
ZnCl.sub.2 (zinc chloride) or zinc bromide, followed by the
addition of an oxalyl chloride mono ester, such as ClCOCOOMe
(methyl chlorooxoacetate) or another ester as above, to afford the
aza-indole glyoxyl ester (Shadrina et al, Ref. 25). Oxalic acid
esters such as methyl oxalate, ethyl oxalate or as above are used.
Aprotic solvents such as CH.sub.2Cl.sub.2, Et.sub.2O, benzene,
toluene, DCE, or the like may be used alone or in combination for
this sequence. In addition to the oxalyl chloride mono esters,
oxalyl chloride itself may be reacted with the azaindole and then
further reacted with an appropriate amine, such as a piperazine
derivative.
[0307] Step C. Hydrolysis of the methyl ester, (intermediates
3a-3e, Schemes 1a-1e) affords a potassium salt of intermediates 4,
which is coupled with N-substituted piperazine derivatives, H--W-A
as shown in Step D of the Schemes 1a-1e. Some typical conditions
employ methanolic or ethanolic sodium hydroxide followed by careful
acidification with aqueous hydrochloric acid of varying molarity
but 1M HCl is preferred. The acidification is not utilized in many
cases as described above for the preferred conditions. Lithium
hydroxide or potassium hydroxide could also be employed and varying
amounts of water could be added to the alcohols. Propanols or
butanols could also be used as solvents. Elevated temperatures up
to the boiling points of the solvents may be utilized if ambient
temperatures do not suffice. Alternatively, the hydrolysis may be
carried out in a non polar solvent such as CH.sub.2Cl.sub.2 or THF
in the presence of Triton B. Temperatures of -78.degree. C. to the
boiling point of the solvent may be employed but -10.degree. C. is
preferred. Other conditions for ester hydrolysis are listed in
reference 41 and both this reference and many of the conditions for
ester hydrolysis are well known to chemists of average skill in the
art.
[0308] Alternative procedures for step B and C:
[0309] Imidazolium Chloroaluminate:
[0310] We found that ionic liquid 1-alkyl-3-alkylimidazolium
chloroaluminate is generally useful in promoting the Friedel-Crafts
type acylation of indoles and azaindoles. The ionic liquid is
generated by mixing 1-alkyl-3-alkylimidazolium chloride with
aluminium chloride at room temperature with vigorous stirring. 1:2
or 1:3 molar ratio of 1-alkyl-3-alkylimidazolium chloride to
aluminium chloride is preferred. One particular useful imidazolium
chloroaluminate for the acylation of azaindole with methyl or ethyl
chlorooxoacetate is the 1-ethyl-3-methylimidazolium
chloroaluminate. The reaction is typically performed at ambient
temperature and the azaindoleglyoxyl ester can be isolated. More
conveniently, we found that the glyoxyl ester can be hydrolyzed in
situ at ambient temperature on prolonged reaction time (typically
overnight) to give the corresponding glyoxyl acid (intermediates
4a-4e) for amide formation (Scheme 2). 23
[0311] A representative experimental procedure is as follows:
l-ethyl-3-methylimidazolium chloride (2 equiv.; purchased from TCI;
weighted under a stream of nitrogen) was stirred in an oven-dried
round bottom flask at r.t. under a nitrogen atmosphere, and added
aluminium chloride (6 equiv.; anhydrous powder packaged under argon
in ampules purchased from Aldrich preferred; weighted under a
stream of nitrogen). The mixture was vigorously stirred to form a
liquid, which was then added azaindole (1 equiv.) and stirred until
a homogenous mixture resulted. The reaction mixture was added
dropwise ethyl or methyl chlorooxoacetate (2 equiv.) and then
stirred at r.t. for 16 h. After which time, the mixture was cooled
in an ice-water bath and the reaction quenched by carefully adding
excess water. The precipitates were filtered, washed with water and
dried under high vacuum to give the azaindoleglyoxylic acid. For
some examples, 3 equivalents of 1-ethyl-3-methylimidazolium
chloride and chlorooxoacetate may be required. A more comprehensive
reference with additional examples is contained in: Yeung, Kap-Sun;
Farkas, Michelle E.; Qiu, Zhilei; Yang, Zhong. Friedel-Crafts
acylation of indoles in acidic imidazolium chloroaluminate ionic
liquid at room temperature. Tetrahedron Letters (2002), 43(33),
5793-5795.
[0312] Related references: (1) Welton, T. Chem Rev. 1999, 99, 2071;
(2) Surette, J. K. D.; Green, L.; Singer, R. D. Chem. Commun. 1996,
2753; (3) Saleh, R. Y. WO 00/15594.
[0313] Step D. Was Described Above.
[0314] It should be noted that in many cases reactions are depicted
for only one position of an intermediate, such as the R.sup.5
position, for example. It is to be understood that such reactions
could be used at other positions, such as R.sup.2-R.sup.4, of the
various intermediates. Reaction conditions and methods given in the
specific examples are broadly applicable to compounds with other
substitution and other tranformations in this application. Schemes
1 and 2 describe general reaction schemes for taking appropriately
substituted Q (indoles and azaindoles) and converting them to
compounds of Formula I. While these schemes are very general, other
permutations such as carrying a precursor or precursors to
substituents R.sup.2 through R.sup.5 through the reaction scheme
and then converting it to a compound of Formula I in the last step
are also contemplated methods of this invention. Nonlimiting
examples of such strategies follow in subsequent schemes.
[0315] The amide bond construction reactions depicted in step D of
schemes 1a-1e could be carried out using the specialized conditions
described herein or alternatively by applying the conditions or
coupling reagents for amide bond construction described in Wallace,
reference 95. Some specific nonlimiting examples are given in this
application.
[0316] Additional procedures for synthesizing, modifying and
attaching groups are contained in references 93-95 and 106 or are
described below. 24
[0317] Scheme 3 provides more specific examples of the
transformation previously described in Schemes A and Schemes 1a-f.
Intermediates 9-13 are prepared by the methodologies as described
for intermediates 1c-5c in Scheme 1c. Scheme 4 is another
embodiment of the transformations described in Schemes 1a-1e and 3.
Conversion of the phenol to the chloride (Step S, Scheme 4) may be
accomplished according to the procedures described in Reimann, E.;
Wichmann, P.; Hoefner, G.; Sci. Pharm. 1996, 64(3), 637-646; and
Katritzky, A. R.; Rachwal, S.; Smith, T. P.; Steel, P. J.; J.
Heterocycl. Chem. 1995, 32(3), 979-984. Step T of Scheme 4 can be
carried out as described for Step A of Scheme 1. The bromo
intermediate can then be converted into alkoxy, chloro, or fluoro
intermediates as shown in Step U of Scheme 4. When step U is the
conversion of the bromide into alkoxy derivatives, the conversion
may be carried out by reacting the bromide with an excess of, for
example, sodium methoxide or potassium methoxide in methanol with
cuprous salts, such as copper I bromide, copper I iodide, and
copper I cyanide. The reaction may be carried out at temperatures
of between ambient and 175.degree. C. but most likely will be
around 115.degree. C. or 100.degree. C. The reaction may be run in
a pressure vessel or sealed tube to prevent escape of volatiles
such as methanol. Alternatively, the reaction can be run in a
solvent such as toluene or xylene and the methanol allowed to
partially escape the reaction vessel by heating and then achieving
reflux by adding a condenser. The preferred conditions on a
typically laboratory scale utilize 3eq of sodium methoxide in
methanol, CuBr as the reaction catalyst (0.2 to 3 equivalents with
the preferred being 1 eq or less), and a reaction temperature of
115.degree. C. The reaction is carried out in a sealed tube or
sealed reaction vessel. The copper catalyzed displacement reaction
of aryl halides by methoxide is described in detail in H. L. Aalten
et al. 1989, Tetrahedron 45(17) pp5565 to 5578 and these conditions
described herein were also utilized in this application with
azaindoles. The conversion of the bromide into alkoxy derivatives
may also be carried out according to procedures described in.
Palucki, M.; Wolfe, J. P.; Buchwald, S. L.; J. Am. Chem. Soc. 1997,
119(14), 3395-3396; Yamato, T.; Komine, M.; Nagano, Y.; Org. Prep.
Proc. Int. 1997, 29(3), 300-303; Rychnovsky, S. D.; Hwang, K.; J.
Org. Chem. 1994, 59(18), 5414-5418. Conversion of the bromide to
the fluoro derivative (Step U, Scheme 4) may be accomplished
according to Antipin, I. S.; Vigalok, A. I.; Konovalov, A. I.; Zh.
Org. Khim. 1991, 27(7), 1577-1577; and Uchibori, Y.; Umeno, M.;
Seto, H.; Qian, Z.; Yoshioka, H.; Synlett. 1992, 4, 345-346.
Conversion of the bromide to the chloro derivative (Step U, Scheme
5) may be accomplished according to procedures described in
Gilbert, E. J.; Van Vranken, D. L.; J. Am. Chem. Soc. 1996,
118(23), 5500-5501; Mongin, F.; Mongin, O.; Trecourt, F.; Godard,
A.; Queguiner, G.; Tetrahedron Lett. 1996, 37(37), 6695-6698; and
O'Connor, K. J.; Burrows, C. J.; J. Org. Chem. 1991, 56(3),
1344-1346. Steps V, W, and X of Scheme 4 are carried out according
to the procedures previously described for Steps B, C, and D of
Scheme 1a-1e, respectively. The steps of Scheme 4 may be carried
out in a different order as shown in Schemes 5 and 6A. 25 26 27
28
[0318] Scheme 6B depicts a shorthand method for depicting the
reactions in Scheme 1a-1e. It is understood, for the purposes of
Scheme 6B and further Schemes, that 1b is used to synthesize 2b-5b,
1c provides 2c-5c and 1d provides 2d-5d etc. The substituents
R.sub.x represent for azaindoles R.sub.2-R.sup.4 and for indoles
R.sub.2-R.sub.5. In formulas in following schemes, one of the
substituents may be depicted but it is understood that each formula
can represent the appropriate generic azaindoles or indole in order
to keep the application succinct.
[0319] An alternative method for carrying out the sequence outlined
in steps B-D (shown in Scheme 6C) involves treating an azaindole,
such as 16, obtained by procedures described in the literature or
from commercial sources, with MeMgI and ZnCl.sub.2, followed by the
addition of ClCOCOCl (oxalyl chloride) in either THF or Et.sub.2O
to afford a mixture of a glyoxyl chloride azaindole, 17a, and an
acyl chloride azaindole, 17b. The resulting mixture of glyoxyl
chloride azaindole and acyl chloride azaindole is then coupled with
mono-benzoylated piperazine derivatives under basic conditions to
afford the products of step D as a mixture of compounds, 18a and
18b, where either one or two carbonyl groups link the azaindole and
group W. Separation via chromatographic methods which are well
known in the art provides the pure 18a and 18b. This sequence is
summarized in Scheme 6C, below. 29 30
[0320] Scheme 6D shows the preparation of an indole intermediate
7a, acylation of 7a with ethyl oxalyl chloride to provide
intermediate 8a, followed by ester hydrolysis to provide
intermediate 9a, and amide formation to provide intermediate
10a.
[0321] Alternatively, the acylation of an indole intermediate, such
as 7a', could be carried out directly with oxalyl chloride followed
by base mediated piperazine coupling to provide an intermediate of
Formula 10a' as shown in Scheme 6E. 31
[0322] Other methods for introduction of an aldehyde group to form
intermediates of formula 11 include transition metal catalyzed
carbonylation reactions of suitable bromo, trifluoromethane
sulfonates(yl), or stannanes(yl) indoles. Alternative the aldehydes
can be introduced by reacting indolyl anions or indolyl Grignard
reagents with formaldehyde and then oxidizing with MnO.sub.2 or
TPAP/NMO or other suitable oxidants to provide intermediate 11.
[0323] Some specific examples of general methods for preparing
functionalized azaindoles or indoles or for interconverting
functionality on aza indoles or indoles which will be useful for
preparing the compounds of this invention are shown in the
following sections for illustrative purposes. It should be
understood that this invention covers substituted 4, 5, 6, and 7
azaindoles and also indoles that the methodology shown below may be
applicable to all of the above series while other shown below will
be specific to one or more. A typical practioner of the art can
make this distinction when not specifically delineated. Many
methods are intended to be applicable to all the series,
particularly functional group installations or interconversions.
For example, a general strategy for providing further functionality
of this invention is to position or install a halide such as bromo,
chloro, or iodo, aldehyde, cyano, or a carboxy group on the
azaindole and then to convert that functionality to the desired
compounds. In particular, conversion to substituted heteroaryl,
aryl, and amide groups on the ring are of particular interest.
[0324] General routes for functionalizing azaindole rings are shown
in Schemes 7, 8 and 9. As depicted in Scheme 7, the azaindole, 17,
can be oxidized to the corresponding N-oxide derivative, 18, by
using mCPBA (meta-Chloroperbenzoic Acid) in acetone or DMF (eq. 1,
Harada et al, Ref. 29 and Antonini et al, Ref. 34). The N-oxide,
18, can be converted to a variety of substituted azaindole
derivatives by using well documented reagents such as phosphorus
oxychloride (POCl.sub.3) (eq. 2, Schneller et al, Ref. 30),
tetramethylammonium fluoride (Me.sub.4NF) (eq. 3), Grignard
reagents RMgX (R=alkyl or aryl, X=Cl, Br or I) (eq. 4, Shiotani et
al, Ref. 31), trimethylsilyl cyanide (TMSCN) (eq. 5, Minakata et
al, Ref. 32) or Ac.sub.2O (eq. 6, Klemm et al, Ref. 33). Under such
conditions, a chlorine (in 19), fluorine (in 20), nitrile (in 22),
alkyl (in 21), aromatic (in 21) or hydroxyl group (in 24) can be
introduced to the pyridine ring. Nitration of azaindole N-oxides
results in introduction of a nitro group to azaindole ring, as
shown in Scheme 8 (eq. 7, Antonini et al, Ref. 34). The nitro group
can subsequently be displaced by a variety of nucleophilic agents,
such as OR, NR.sup.1R.sup.2 or SR, in a well established chemical
fashion (eq. 8, Regnouf De Vains et al, Ref. 35(a), Miura et al,
Ref. 35(b), Profft et al, Ref. 35(c)). The resulting N-oxides, 26,
are readily reduced to the corresponding azaindole, 27, using
phosphorus trichloride (PCl.sub.3) (eq. 9, Antonini et al, Ref 0.34
and Nesi et al, Ref. 36). Similarly, nitro-substituted N-oxide, 25,
can be reduced to the azaindole, 28, using phosphorus trichloride
(eq. 10). The nitro group of compound 28 can be reduced to either a
hydroxylamine (NHOH), as in 29, (eq. 11, Walser et al, Ref. 37(a)
and Barker et al, Ref. 37(b)) or an amino (NH.sub.2) group, as in
30, (eq. 12, Nesi et al, Ref. 36 and Ayyangar et al, Ref. 38) by
carefully selecting different reducing conditions. 3233 3435
[0325] The alkylation of the nitrogen atom at position 1 of the
azaindole derivatives can be achieved using NaH as the base, DMF as
the solvent and an alkyl halide or sulfonate as alkylating agent,
according to a procedure described in the literature (Mahadevan et
al, Ref. 39) (Scheme 9). 36
[0326] In the general routes for substituting the azaindole ring
described above, each process can be applied repeatedly and
combinations of these processes is permissible in order to provide
azaindoles incorporating multiple substituents. The application of
such processes provides additional compounds of Formula I. 37
[0327] The synthesis of 4-aminoazaindoles which are useful
precursors for 4, 5, and/or 7-substituted azaindoles is shown in
Scheme 10 above.
[0328] The synthesis of 3,5-dinitro-4-methylpyridine, 32, is
described in the following two references by Achremowicz et. al.:
Achremowicz, Lucjan. Pr. Nauk. Inst. Chem. Org. Fiz. Politech.
Wroclaw. 1982, 23, 3-128; Achremowicz, Lucjan. Synthesis 1975, 10,
653-4. In the first step of Scheme 10, the reaction with
dimethylformamide dimethyl acetal in an inert solvent or neat under
conditions for forming Batcho-Leimgruber precursors provides the
cyclization precursor, 33, as shown. Although the step is
anticipated to work as shown, the pyridine may be oxidized to the
N-oxide prior to the reaction using a peracid such as MCPBA or a
more potent oxidant like meta-trifluoromethyl or meta nitro peroxy
benzoic acids. In the second step of Scheme 10, reduction of the
nitro group using for example hydrogenation over Pd/C catalyst in a
solvent such as MeOH, EtOH, or EtOAc provides the cyclized product,
34. Alternatively the reduction may be carried out using tin
dichloride and HCl, hydrogenation over Raney nickel or other
catalysts, or by using other methods for nitro reduction such as
described elsewhere in this application. A general method for
preparing indoles and azaindoles of the invention utilize the
Leim-Gruber Batcho-reation sequence as shown in the scheme below:
38
[0329] The amino indole, 34, can now be converted to compounds of
Formula I via, for example, diazotization of the amino group, and
then conversion of the diazonium salt to the fluoride, chloride or
alkoxy group. See the discussion of such conversions in the
descriptions for Schemes 17 and 18. The conversion of the amino
moiety into desired functionality could then be followed by
installation of the oxoacetopiperazine moiety by the standard
methodology described above. 5 or 7-substitution of the azaindole
can arise from N-oxide formation at position 6 and subsequent
conversion to the chloro via conditions such as POCl.sub.3 in
chloroform, acetic anhydride followed by POCl.sub.3 in DMF, or
alternatively TsCl in DMF. Literature references for these and
other conditions are provided in some of the later Schemes in this
application. The synthesis of 4-bromo-7-hydroxy or protected
hydroxy-4-azaindole is described below as this is a useful
precursor for 4 and/or 7 substituted 6-aza indoles.
[0330] The synthesis of 5-bromo-2-hydroxy-4-methyl-3-nitro
pyridine, 35, may be carried out as described in the following
reference: Betageri, R.; Beaulieu, P. L.; Llinas-Brunet, M;
Ferland, J. M.; Cardozo, M.; Moss, N.; Patel, U.; Proudfoot, J. R.
PCT Int. Appl. WO 9931066, 1999. Intermediate 36 is prepared from
35 according to the method as described for Step 1 of Scheme 11. PG
is an optional hydroxy protecting group such as triallylsilyl,
methyl, benzyl or the like. Intermediate 37 is then prepared from
36 by the selective reduction of the nitro group in the presence of
bromide and subsequent cyclization as described in the second step
of Scheme 10. Fe(OH).sub.2 in DMF with catalytic tetrabutylammonium
bromide can also be utilized for the reduction of the nitro group.
The bromide may then be converted to alkoy using the conditions
employed in step U of scheme 4. The compounds are then converted to
compounds of Formula I as above. The protecting group on the C-7
position may be removed with TMSI, hydrogenation orin the case of
allyl standard palladium deprotection conditions in order to
generate the free C-7 hydroxy compound which can also be depicted
as its pyridone tautomer. As described earlier POBr3 or POCl.sub.3
can be used to convert the hydroxy intermediate to the C-7 bromo or
chloro intermediate respectively. 39
[0331] Step E. Scheme 14 depicts the nitration of an azaindole, 41,
(R.sub.2.dbd.H). Numerous conditions for nitration of the azaindole
may be effective and have been described in the literature.
N.sub.2O.sub.5 in nitromethane followed by aqueous sodium bisulfite
according to the method of Bakke, J. M.; Ranes, E.; Synthesis 1997,
3, 281-283 could be utilized. Nitric acid in acetic may also be
employed as described in Kimura, H.; Yotsuya, S.; Yuki, S.; Sugi,
H.; Shigehara, I.; Haga, T.; Chem. Pharm. Bull. 1995, 43(10),
1696-1700. Sulfuric acid followed by nitric acid may be employed as
in Ruefenacht, K.; Kristinsson, H.; Mattern, G.; Helv Chim Acta
1976, 59, 1593. Coombes, R. G.; Russell, L. W.; J. Chem. Soc.,
Perkin Trans. 1 1974, 1751 describes the use of a Titatanium based
reagent system for nitration. Other conditions for the nitration of
the azaindole can be found in the following references: Lever, O.
W. J.; Werblood, H. M.; Russell, R. K.; Synth. Comm. 1993, 23(9),
1315-1320; Wozniak, M.; Van Der Plas, H. C.; J. Heterocycl Chem.
1978, 15, 731. 40
[0332] Step F. As shown above in Scheme 15, Step F, substituted
azaindoles containing a chloride, bromide, iodide, triflate, or
phosphonate undergo coupling reactions with a boronate (Suzuki type
reactions) or a stannane (Stille type coupling) to provide
substituted indoles or azaindoles. This type of coupling as
mentioned previously can also be used to functionalize vinyl
halides, triflates or phosphonates to add groups D or A or
precursors. Stannanes and boronates are prepared via standard
literature procedures or as described in the experimental section
of this application. The substitututed indoles, azaindoles, or
alkenes may undergo metal mediated coupling to provide compounds of
Formula I wherein R.sup.4 is aryl, heteroaryl, or heteroalicyclic
for example. The indoles or azaindole intermediates, (halogens,
triflates, phosphonates) may undergo Stille-type coupling with
heteroarylstannanes as shown in Scheme 15 or with the corresponding
vinyl reagents as described in earlier Schemes. Conditions for this
reaction are well known in the art and the following are three
example references a) Farina, V.; Roth, G. P. Recent advances in
the Stille reaction; Adv. Met.-Org. Chem. 1996, 5, 1-53. b) Farina,
V.; Krishnamurthy, V.; Scott, W. J. The Stille reaction; Org.
React. (N.Y.) 1997, 50, 1-652. and c) Stille, J. K. Angew. Chem.
Int. Ed. Engl. 1986, 25, 508-524. Other references for general
coupling conditions are also in the reference by Richard C. Larock
Comprehensive Organic Transformations 2nd Ed. 1999, John Wiley and
Sons New York. All of these references provide numerous conditions
at the disposal of those skilled in the art in addition to the
specific examples provided in Scheme 15 and in the specific
embodiments. It can be well recognized that an indole stannane
could also couple to a heterocyclic or aryl halide or triflate to
construct compounds of Formula I. Suzuki coupling (Norio Miyaura
and Akiro Suzuki Chem Rev. 1995, 95, 2457.) between a triflate,
bromo, or chloro azaindole intermediate and a suitable boronate
could also be employed and some specific examples are contained in
this application. Palladium catalyzed couplings of stannanes and
boronates between halo azaindole or indole intermediates or vinyl
halides or vinyl triflates or similar vinyl substrate are also
feasible and have been utilized extensively for this invention.
Preferred procedures for coupling of a chloro or bromo azaindole or
vinyl halide and a stannane employ dioxane, stoichiometric or an
excess of the tin reagent (up to 5 equivalents), 0.1 to 1 eq of
tetrakis triphenyl phosphine Palladium (O) in dioxane heated for 5
to 15 h at 110 to 1200. Other solvents such as DMF, THF, toluene,
or benzene could be employed. Another useful procedure for coupling
a halo indole or azaindole with a suitable tributyl heteroaryl or
other stannane employs usually a slight excess (1.1 eqs) but up to
several equivalents of the stannane, 0.1 eqs CuI, 0.1 equivalents
of tetrakis triphenyl phosphine palladium (0) all of which is
usually dissolved in dry DMF (approximately 5 mmol of halide per 25
mL of DMF but this concentration can be reduced for sluggish
reactions or increased if solubility is an issue). The reaction is
usually heated at an elevated temperature of about 90.degree. C.
and the reaction is usually run in a sealed reaction vessel or
sealed tube. When the reaction is completed it is usually allowed
to cool, filtered through methanesulfonic acid SCX cartridges with
MeOH to remove triphenyl phosphine oxide, and then purified by
standard crystallization or chromatographic methods. Examples of
the utility of these conditions are shown in Scheme Z below. 41
[0333] Alternatively, the Stille type coupling between a stannane
(.about.1.1 eqs) and a vinyl, heteroaryl, or aryl halide may
proceed better using (0.05 to 0.1 eq) bvPd2(dba).sub.3 as catalyst
and tri-2-furylphosphine (.about.0.25eq) as the added ligand. The
reaction is usually heated in THF or dioxane at a temperature
between 70 and 90.degree. C. Preferred procedures for Suzuki
coupling of a chloro azaindole and a boronate employ 1:1 DMF water
as solvent, 2 equivalents of potassium carbonate as base
stoichiometric or an excess of the boron reagent (up to 5
equivalents), 0.1 to 1 eq of Palladium (O) tetrakis triphenyl
phosphine heated for 5 to 15 h at 110 to 1200. Less water is
occasionally employed. Another useful condition for coupling a
heteroaryl or aryl boronic acid to a stoichiometric amount of vinyl
halide or triflate utilizes DME as solvent (.about.0.33 mmol halide
per 3 mL DME), .about.4eq of 2M sodium carbonate, and 0.05 eq Pd2
dba3 heated in a sealed tube or sealed vessel at 90.degree. C. for
.about.16 h. Reaction times vary with substrate. Another useful
method for coupling involves use of coupling an aryl, heteroaryl or
vinyl zinc bromide or chloride coupled with a vinyl, aryl or
heteroaryl halide using tetrakis triphenyl phosphine palladium (O)
heated in THF. Detailed example procedures for preparing the zinc
reagents from halides via lithium bromide exhange and then
transmetalation and reaction conditions are contained in the
experimental section. If standard conditions fail new specialized
catalysts and conditions can be employed. Discussions on details,
conditions, and alternatives for carrying out the metal mediated
couplings described above can also be found in the book
"Organometallics in Organic Synthesis; A Manual; 2002, 2.sup.nd Ed.
M. Schlosser editor, John Wiley and Sons, West Sussex, England,
ISBN 0 471 98416 7.
[0334] Some references (and the references therein) describing
catalysts which are useful for coupling with aryl and heteroaryl
chlorides are:
[0335] Littke, A. F.; Dai, C.; Fu, G. C. J. Am. Chem. Soc. 2000,
122(17), 4020-4028; Varma, R. S.; Naicker, K. P. Tetrahedron Lett.
1999, 40(3), 439-442; Wallow, T. I.; Novak, B. M. J. Org. Chem.
1994, 59(17), 5034-7; Buchwald, S.; Old, D. W.; Wolfe, J. P.;
Palucki, M.; Kamikawa, K.; Chieffi, A.; Sadighi, J. P.; Singer, R.
A.; Ahman, J PCT Int. Appl. WO 0002887 2000; Wolfe, J. P.;
Buchwald, S. L. Angew. Chem., Int. Ed. 1999, 38(23), 3415; Wolfe,
J. P.; Singer, R. A.; Yang, B. H.; Buchwald, S. L. J. Am. Chem.
Soc. 1999,121(41), 9550-9561; Wolfe, J. P.; Buchwald, S. L. Angew.
Chem., Int. Ed. 1999, 38(16), 2413-2416; Bracher, F.; Hildebrand,
D.; Liebigs Ann. Chem. 1992, 12, 1315-1319; and Bracher, F.;
Hildebrand, D.; Liebigs Ann. Chem. 1993, 8, 837-839.
[0336] Alternatively, the boronate or stannane may be formed on the
azaindole via methods known in the art and the coupling performed
in the reverse manner with aryl or heteroaryl based halogens or
triflates.
[0337] Known boronate or stannane agents could be either purchased
from commercial resources or prepared following disclosed
documents. Additional examples for the preparation of tin reagents
or boronate reagents are contained in the experimental section, and
references 93-95 and 106.
[0338] Novel stannane agents could be prepared from one of the
following routes which should not be viewed as limiting. 42 43 44
45 46
[0339] Boronate reagents are prepared as described in reference 71.
Reaction of lithium or Grignard reagents with trialkyl borates
generates boronates. Alternatively, Palladium catalyzed couplings
of alkoxy diboron or alkyl diboron reagents with aryl or heteroaryl
halides can provide boron reagents for use in Suzuki type
couplings. Some example conditions for coupling a halide with
(MeO)BB(OMe).sub.2 utilize PdCl2 (dppf), KOAc, DMSO, at 80.degree.
C. until reaction is complete when followed by TLC or HPLC
analysis.
[0340] Related examples are provided in the following experimental
section.
[0341] Methods for direct addition of aryl or heteroaryl
organometallic reagents to alpha chloro nitrogen containing
heterocyles or the N-oxides of nitrogen containing heterocycles are
known and applicable to the azaindoles. Some examples are Shiotani
et. Al. J. Heterocyclic Chem. 1997, 34(3), 901-907; Fournigue et.
al. J. Org. Chem. 1991, 56(16), 4858-4864. 47 48
[0342] As shown in Schemes 12 and 13, a mixture of halo-indole or
halo-azaindole intermediate, 1-2 equivalents of copper powder, with
1 equivalent preferred for the 4-F,6-azaindole series and 2
equivalents for the 4-methoxy,6-azaindole series; 1-2 equivalents
of potassium carbonate, with 1 equivalent preferred for the
4-F,6-azaindole series and 2 equivalents for the
4-methoxy,6-azaindole series; and a 2-30 equivalents of the
corresponding heterocyclic reagent, with 10 equivalents preferred;
was heated at 135-160.degree. C. for 4 to 9 hours, with 5 hours at
160.degree. C. preferred for the 4-F,6-azaindole series and 7 hours
at 135.degree. C. preferred for the 4-methoxy,6-azaindole series.
The reaction mixture was cooled to room temperature and filtered
through filter paper. The filtrate was diluted with methanol and
purified either by preparative HPLC or silica gel. In many cases no
chromatography is necessary, the product can be obtained by
crystallization with methanol.
[0343] Alternatively, the installation of amines or N linked
heteroaryls may be carried out by heating 1 to 40 equivalents of
the appropriate amine and an equivalent of the appropriate aza
indole chloride, bromide or iodide with copper bronze (from 0.1 to
10equivalents (preferably about 2 equivalents) and from 1 to 10
equivalents of finely pulverized potassium hydroxide (preferably
about 2 equivalents). Temperatures of 120.degree. to 2000 may be
employed with 140-1600 generally preferred. For volatile starting
materials a sealed reactor may be employed. The reaction is most
commonly used when the halogen being displaced is at the 7-position
of a 6-aza or 4-azaindole but the method can work in the
5-azaseries or when the halogen is at a different position (4-7
position possible). As shown above the reaction could be employed
on azaindoles unsubstituted at position 3 or intermediates which
contain the dicarbonyl or the intact dicarbonyl N-heteroaryl
piperazine. 49
[0344] A possible preparation of a key aldehyde intermediate, 43,
using a procedure adapted from the method of Gilmore et. Al.
Synlett 1992, 79-80, is shown in Scheme 16 above. The aldehyde
substituent is shown only at the R.sub.4 position for the sake of
clarity, and should not be considered as a limitation of the
methodology. The bromide or iodide intermediate is converted into
an aldehyde intermediate, 43, by metal-halogen exchange and
subsequent reaction with dimethylformamide in an appropriate
aprotic solvent. Typical bases which could be used include, but are
not limited to, alkyl lithium bases such as n-butyl lithium, sec
butyl lithium or tert butyl lithium or a metal such as lithium
metal. A preferred aprotic solvent is THF. Typically the
transmetallation is initiated at -78.degree. C. The reaction may be
allowed to warm to allow the transmetalation to go to completion
depending on the reactivity of the bromide intermediate. The
reaction is then recooled to -78.degree. C. and allowed to react
with dimethylformamide (allowing the reaction to warm may be
required to enable complete reaction) to provide an aldehyde which
is elaborated to compounds of Formula I. Other methods for
introduction of an aldehyde group to form intermediates of formula
43 include transition metal catalyzed carbonylation reactions of
suitable bromo, trifluoromethane sulfonyl, or stannyl azaindoles.
Alternatively the aldehydes could be introduced by reacting indolyl
anions or indolyl Grignard reagents with formaldehyde and then
oxidizing with MnO.sub.2 or TPAP/NMO or other suitable oxidants to
provide intermediate 43.
[0345] The methodology described in T. Fukuda et. al. Tetrahedron
1999, 55, 9151 and M. Iwao et. Al. Heterocycles 1992, 34(5), 1031
provide methods for preparing indoles with substituents at the
7-position. The Fukuda references provide methods for
functionalizing the C-7 position of indoles by either protecting
the indole nitrogen with 2,2-diethyl propanoyl group and then
deprotonating the 7-position with sec/Buli in TMEDA to give an
anion. This anion may be quenched with DMF, formaldehyde, or carbon
dioxide to give the aldehyde, benzyl alcohol, or carboxylic acid
respectively and the protecting group removed with aqueous t
butoxide. Similar tranformations could be achieved by converting
indoles to indoline, lithiation at C-7 and then reoxidation to the
indole such as described in the Iwao reference above. The oxidation
level of any of these products may be adjusted by methods well
known in the art as the interconversion of alcohol, aldehyde, and
acid groups has been well studied. It is also well understood that
a cyano group can be readily converted to an aldehyde. A reducing
agent such as DIBALH in hexane such as used in Weyerstahl, P.;
Schlicht, V.; Liebigs Ann/Recl. 1997, 1, 175-177 or alternatively
catecholalane in THF such as used in Cha, J. S.; Chang, S. W.;
Kwon, O. O.; Kim, J. M.; Synlett. 1996, 2, 165-166 will readily
achieve this conversion to provide intermediates such as 44 (Scheme
16). Methods for synthesizing the nitriles are shown later in this
application. It is also well understood that a protected alcohol,
aldehyde, or acid group could be present in the starting azaindole
and carried through the synthetic steps to a compound of Formula I
in a protected form until they can be converted into the desired
substituent at R.sup.1 through R.sup.4. For example, a benzyl
alcohol can be protected as a benzyl ether or silyl ether or other
alcohol protecting group; an aldehyde may be carried as an acetal,
and an acid may be protected as an ester or ortho ester until
deprotection is desired and carried out by literature methods.
50
[0346] Step G. Step 1 of Scheme 17 shows the reduction of a nitro
group on 45 to the amino group of 46. Although shown on position 4
of the azaindole, the chemistry is applicable to other nitro
isomers. The procedure described in Ciurla, H.; Puszko, A.; Khim
Geterotsikl Soedin 1996, 10, 1366-1371 uses hydrazine Raney-Nickel
for the reduction of the nitro group to the amine. Robinson, R. P.;
DonahueO, K. M.; Son, P. S.; Wagy, S. D.; J. Heterocycl. Chem.
1996, 33(2), 287-293 describes the use of hydrogenation and Raney
Nickel for the reduction of the nitro group to the amine. Similar
conditions are described by Nicolai, E.; Claude, S.; Teulon, J. M.;
J. Heterocycl. Chem. 1994, 31(1), 73-75 for the same
transformation. The following two references describe some
trimethylsilyl sulfur or chloride based reagents which may be used
for the reduction of a nitro group to an amine. Hwu, J. R.; Wong,
F. F.; Shiao, M. J.; J. Org. Chem. 1992, 57(19), 5254-5255; Shiao,
M. J.; Lai, L. L.; Ku, W. S.; Lin, P. Y.; Hwu, J. R.; J. Org. Chem.
1993, 58(17), 4742-4744.
[0347] Step 2 of Scheme 17 describes general methods for conversion
of amino groups on azaindoles or indoles into other functionality.
Scheme 18 also depicts transformations of an amino azaindole into
various intermediates and compounds of Formula I.
[0348] The amino group at any position of the azaindole, such as 46
(Scheme 17), could be converted to a hydroxy group using sodium
nitrite, sulfuric acid, and water via the method of Klemm, L. H.;
Zell, R.; J. Heterocycl. Chem. 1968, 5, 773. Bradsher, C. K.;
Brown, F. C.; Porter, H. K.; J. Am. Chem. Soc. 1954, 76, 2357
describes how the hydroxy group may be alkylated under standard or
Mitsonobu conditions to form ethers. The amino group may be
converted directly into a methoxy group by diazotization (sodium
nitrite and acid) and trapping with methanol.
[0349] The amino group of an azaindole, such as 46, could be
converted to fluoro via the method of Sanchez using HPF.sub.6,
NaNO.sub.2, and water by the method described in Sanchez, J. P.;
Gogliotti, R. D.; J. Heterocycl. Chem. 1993, 30(4), 855-859. Other
methods useful for the conversion of the amino group to fluoro are
described in Rocca, P.; Marsais, F.; Godard, A.; Queguiner, G.;
Tetrahedron Lett. 1993, 34(18), 2937-2940 and Sanchez, J. P.;
Rogowski, J. W.; J. Heterocycl. Chem. 1987, 24, 215.
[0350] The amino group of the azaindole, 46, could also be
converted to a chloride via diazotization and chloride displacement
as described in Ciurla, H.; Puszko, A.; Khim Geterotsikl Soedin
1996, 10, 1366-1371 or the methods in Raveglia, L. F.; Giardina, G.
A.; Grugni, M.; Rigolio, R.; Farina, C.; J. Heterocycl. Chem. 1997,
34(2), 557-559 or the methods in Matsumoto, J. I.; Miyamoto, T.;
Minamida, A.; Mishimura, Y.; Egawa, H.; Mishimura, H.; J. Med.
Chem. 1984, 27(3), 292; or as in Lee, T. C.; Salemnick, G.; J. Org.
Chem. 1975, 24, 3608.
[0351] The amino group of the azaindole, 46, could also be
converted to a bromide via diazotization and displacement by
bromide as described in Raveglia, L. F.; Giardina, G. A.; Grugni,
M.; Rigolio, R.; Farina, C.; J. Heterocycl. Chem. 1997, 34(2),
557-559; Talik, T.; Talik, Z.; Ban-Oganowska, H.; Synthesis 1974,
293; and Abramovitch, R. A.; Saha, M.; Can. J. Chem. 1966, 44,
1765. 51
[0352] The preparation of 4-amino 4-azaindole and
7-methyl-4-azaindole is described by Mahadevan, I.; Rasmussen, M.
J. Heterocycl. Chem. 1992, 29(2), 359-67. The amino group of the
4-amino 4-azaindole can be converted to halogens, hydroxy,
protected hydroxy, triflate, as described above in Schemes 17-18
for the 4-amino compounds or by other methods known in the art.
Protection of the indole nitrogen of the 7-methyl-4-azaindole via
acetylation or other strategy followed by oxidation of the 7-methyl
group with potassium permanganate or chromic acid provides the
7-acid/4-N-oxide. Reduction of the N-oxide, as described below,
provides an intermediate from which to install various substituents
at position R.sub.4. Alternatively the parent 4-azaindole which was
prepared as described in Mahadevan, I.; Rasmussen, M. J.
Heterocycl. Chem. 1992, 29(2), 359-67 could be derivatized at
nitrogen to provide the 1-(2,2-diethylbutanoyl)azaindole which
could then be lithiated using TMEDA/sec BuLi as described in T.
Fukuda et. Al. Tetrahedron 1999, 55, 9151-9162; followed by
conversion of the lithio species to the 7-carboxylic acid or
7-halogen as described. Hydrolysis of the N-amide using aqueous
tert-butoxide in THF regenerates the free NH indole which could
then be converted to compounds of Formula I. The chemistry used to
functionalize position 7 can also be applied to the 5 and 6 indole
series.
[0353] Scheme 19 shows the preparation of a 7-chloro-4-azaindole,
50, which could be converted to compounds of Formula I by the
chemistry previously described, especially the palladium catalyzed
tin and boron based coupling methodology described above. The
chloro nitro indole, 49, is commercially available or can be
prepared from 48 according to the method of Delarge, J.; Lapiere,
C. L. Pharm. Acta Helv. 1975, 50(6), 188-91. 52
[0354] Scheme 20, below, shows another synthetic route to
substituted 4-aza indoles. The 3-aminopyrrole, 51, was reacted to
provide the pyrrolopyridinone, 52, which was then reduced to give
the hydroxy azaindole, 53. The pyrrolo[2,3-b]pyridines described
were prepared according to the method of Britten, A. Z.; Griffiths,
G. W. G. Chem. Ind. (London) 1973, 6, 278. The hydroxy azaindole,
53, could then be converted to the triflate then further reacted to
provide compounds of Formula I. 53
[0355] The following references describe the synthesis of 7-halo or
7 carboxylic acid, or 7-amido derivatives of 5-azaindoline which
can be used to construct compounds of Formula I. Bychikhina, N. N.;
Azimov, V. A.; Yakhontov, L. N. Khim. Geterotsikl. Soedin. 1983, 1,
58-62; Bychikhina, N. N.; Azimov, V. A.; Yakhontov, L. N. Khim.
Geterotsikl. Soedin. 1982, 3, 356-60; Azimov, V. A.; Bychikhina, N.
N.; Yakhontov, L. N. Khim. Geterotsikl. Soedin. 1981, 12, 1648-53;
Spivey, A. C.; Fekner, T.; Spey, S. E.; Adams, H. J. Org. Chem.
1999, 64(26), 9430-9443; Spivey, A. C.; Fekner, T.; Adams, H.
Tetrahedron Lett. 1998, 39(48), 8919-8922. The methods described in
Spivey et al. (preceding two references) for the preparation of
1-methyl-7-bromo-4-azaindoline can be used to prepare the
1-benzyl-7-bromo-4-azaindoline, 54, shown below in Scheme 21. This
could be utilized in Stille or Suzuki couplings to provide 55,
which is deprotected and dehydrogenated to provide 56. Other useful
azaindole intermediates, such as the cyano derivatives, 57 and 58,
and the aldehyde derivatives, 59 and 60, can then be further
elaborated to compounds of Formula I. 54
[0356] Alternatively the 7-functionalized 5-azaindole derivatives
could be obtained by functionalization using the methodologies of
T. Fukuda et. al. Tetrahedron 1999, 55, 9151 and M. Iwao et. Al.
Heterocycles 1992, 34(5), 1031 described above for the 4 or 6
azaindoles. The 4 or 6 positions of the 5 aza indoles can be
functionalized by using the azaindole N-oxide.
[0357] The conversion of indoles to indolines is well known in the
art and can be carried out as shown or by the methods described in
Somei, M.; Saida, Y.; Funamoto, T.; Ohta, T. Chem. Pharm. Bull.
1987, 35(8), 3146-54; M. Iwao et. Al. Heterocycles 1992, 34(5),
1031; and Akagi, M.; Ozaki, K. Heterocycles 1987, 26(1), 61-4.
55
[0358] The preparation of azaindole oxoacetyl or oxo piperidines
with carboxylic acids could be carried out from nitrile, aldehyde,
or anion precursors via hydrolysis, oxidation, or trapping with
CO.sub.2 respectively. As shown in the Scheme 22, Step 1, or the
scheme below step a12 one method for forming the nitrile
intermediate, 62, is by cyanide displacement of a halide in the
aza-indole ring. The cyanide reagent used can be sodium cyanide, or
more preferably copper or zinc cyanide. The reactions could be
carried out in numerous solvents which are well known in the art.
For example DMF is used in the case of copper cyanide. Additional
procedures useful for carrying out step 1 of Scheme 24 are
Yamaguchi, S.; Yoshida, M.; Miyajima, I.; Araki, T.; Hirai, Y.; J.
Heterocycl. Chem. 1995, 32(5), 1517-1519 which describes methods
for copper cyanide; Yutilov, Y. M.; Svertilova, I. A.; Khim
Geterotsikl Soedin 1994, 8, 1071-1075 which utilizes potassium
cyanide; and Prager, R. H.; Tsopelas, C.; Heisler, T.; Aust. J.
Chem. 1991, 44 (2), 277-285 which utilizes copper cyanide in the
presence of MeOS(O).sub.2F. The chloride or more preferably a
bromide on the azaindole could be displaced by sodium cyanide in
dioxane via the method described in Synlett. 1998, 3, 243-244.
Alternatively, Nickel dibromide, Zinc, and triphenyl phosphine in
can be used to activate aromatic and heteroaryl chlorides to
displacement via potassium cyanide in THF or other suitable solvent
by the methods described in Eur. Pat. Appl., 831083, 1998.
[0359] The conversion of the cyano intermediate, 62, to the
carboxylic acid intermediate, 63, is depicted in step 2, Scheme 22
or in step a12, Scheme 23. Many methods for the conversion of
nitrites to acids are well known in the art and may be employed.
Suitable conditions for step 2 of Scheme 22 or the conversion of
intermediate 65 to intermediate 66 below employ potassium
hydroxide, water, and an aqueous alcohol such as ethanol. Typically
the reaction must be heated at refluxing temperatures for one to
100 h. Other procedures for hydrolysis include those described
in:
[0360] Shiotani, S.; Taniguchi, K.; J. Heterocycl. Chem. 1997,
34(2), 493-499; Boogaard, A. T.; Pandit, U. K.; Koomen, G.-J.;
Tetrahedron 1994, 50(8), 2551-2560; Rivalle, C.; Bisagni, E.;
Heterocycles 1994, 38(2), 391-397; Macor, J. E.; Post, R.; Ryan,
K.; J. Heterocycl. Chem. 1992, 29(6), 1465-1467.
[0361] The acid intermediate, 66 (Scheme 23), could then be
esterified using conditions well known in the art. For example,
reaction of the acid with diazomethane in an inert solvent such as
ether, dioxane, or THF would give the methyl ester. Intermediate 67
may then be converted to intermediate 68 according to the procedure
described in Scheme 2. Intermediate 68 could then be hydrolyzed to
provide intermediate 69. 56
[0362] As shown in Scheme 24, step a13 another preparation of the
indoleoxoacetylpiperazine 7-carboxylic acids, 69, is carried out by
oxidation of the corresponding 7-carboxaldehyde, 70. Numerous
oxidants are suitable for the conversion of aldehyde to acid and
many of these are described in standard organic chemistry texts
such as: Larock, Richard C., Comprehensive organic transformations:
a guide to functional group preparations 2.sup.nd ed. New York:
Wiley-VCH, 1999. One preferred method is the use of silver nitrate
or silver oxide in a solvent such as aqueous or anhydrous methanol
at a temperature of 25.degree. C. or as high as reflux. The
reaction is typically carried out for one to 48 h and is typically
monitored by TLC or LC/MS until complete conversion of product to
starting material has occurred. Alternatively, KmnO.sub.4 or
CrO.sub.3/H.sub.2SO.sub.4 could be utilized. 57
[0363] Scheme 25 gives a specific example of the oxidation of an
aldehyde intermediate, 70a, which could be used to provide the
carboxylic acid intermediate, 69a. 58
[0364] Alternatively, intermediate 69 could be prepared by the
nitrile method of synthesis carried out in an alternative order as
shown in Scheme 26. The nitrile hydrolyis step can be delayed and
the nitrile carried through the synthesis to provide a nitrile
which could be hydrolyzed to provide the free acid, 69, as above.
59 60
[0365] Step H. The direct conversion of nitriles, such as 72, to
amides, such as 73, shown in Scheme 27, Step H, could be carried
out using the conditions as described in Shiotani, S.; Taniguchi,
K.; J. Heterocycl. Chem. 1996, 33(4), 1051-1056 (describes the use
of aqueous sulfuric acid); Memoli, K. A.; Tetrahedron Lett. 1996,
37(21), 3617-3618; Adolfsson, H.; Waemmark, K.; Moberg, C.; J. Org.
Chem. 1994, 59(8), 2004-2009; and El Hadri, A.; Leclerc, G.; J.
Heterocycl. Chem. 1993, 30(3), 631-635.
[0366] Step I. for NH2
[0367] Shiotani, S.; Taniguchi, K.; J. Heterocycl. Chem. 1997,
34(2), 493-499; Boogaard, A. T.; Pandit, U. K.; Koomen, G.-J.;
Tetrahedron 1994, 50(8), 2551-2560; Rivalle, C.; Bisagni, E.;
Heterocycles 1994, 38(2), 391-397; Macor, J. E.; Post, R.; Ryan,
K.; J. Heterocycl. Chem. 1992, 29(6), 1465-1467.
[0368] Step J. 61
[0369] The following scheme (28A) shows an example for the
preparation of 4-fluoro-7substituted azaindoles from a known
starting materials. References for the Bartoli indole synthesis
were mentioned earlier. The conditions for tranformation to the
nitrites, acids, aldeheydes, heterocycles and amides have also been
described in this application. 62 63
[0370] Steps a16, a17, and a18 encompasses reactions and conditions
for 1.sup.0, 2.sup.0 and 3.sup.0 amide bond formation as shown in
Schemes 28 and 29 which provide compounds such as those of Formula
73.
[0371] The reaction conditions for the formation of amide bonds
encompass any reagents that generate a reactive intermediate for
activation of the carboxylic acid to amide formation, for example
(but not limited to), acyl halide, from carbodiimide, acyl iminium
salt, symmetrical anhydrides, mixed anhydrides (including
phosphonic/phosphinic mixed anhydrides), active esters (including
silyl ester, methyl ester and thioester), acyl carbonate, acyl
azide, acyl sulfonate and acyloxy N-phosphonium salt. The reaction
of the indole carboxylic acids with amines to form amides may be
mediated by standard amide bond forming conditions described in the
art. Some examples for amid bond formation are listed in references
41-53 but this list is not limiting. Some carboxylic acid to amine
coupling reagents which are applicable are EDC,
Diisopropylcarbodiimide or other carbodiimides, PyBop
(benzotriazolyloxytris(dimethylamino) phosphonium
hexafluorophosphate), 2-(1H-benzotriazole-1-yl)-1, 1, 3,
3-tetramethyl uronium hexafluorophosphate (HBTU). A particularly
useful method for azaindole 7-carboxylic acid to amide reactions,
likely to be based on an analogous series, is the use of carbonyl
imidazole as the coupling reagent as described in reference 53. The
temperature of this reaction could be lower than in the cited
reference, from 80.degree. C. (or possibly lower) to 150.degree. C.
or higher. A more specific possible application in which W is
piperazinyl is depicted in Scheme 30. 64
[0372] The following four general methods provide a more detailed
description for the preparation of indolecarboamides and these
methods were employed for the synthesis of compounds similar to
Formula I, except that A formed a carboxamide. These methods should
work as written to provide compounds of Formula I.
[0373] Method 1:
[0374] To a mixture of an acid intermediate, such as 75, (1 equiv),
an appropriate amine (4 equiv.) and DMAP 0.1 tp 1 eq would be
dissolved CH.sub.2Cl.sub.2 (1 mL) and then EDC added (1 eq). The
resulting mixture should be shaken at rt for 12 h, and then
evaporated in vacuo. The residue could be dissolved in a solvent
such as MeOH, and subjected to preparative reverse phase HPLC
purification.
[0375] Method 2:
[0376] To a mixture of an appropriate amine (4 equiv.) and HOBT (16
mg, 0.12 mmol) in THF (0.5 mL) should be added an acid
intermediate, such as 74, and NMM 1 eq followed by EDC. The
reaction mixture could be shaken at rt for 12 h. The volatiles
should be evaporated in vacuo; and the residue dissolved in MeOH
and subjected to preparative reverse phase HPLC purification.
[0377] Method 3:
[0378] To a mixture of an acid intermediate, such as 74, amine (4
equiv.) and DEPBT (prepared according to Li, H.; Jiang, X. Ye, Y.;
Fan, C.; Todd, R.; Goodman, M. Organic Letters 1999, 1, 91); in DMF
would be added TEA. The resulting mixture should be shaken at rt
for 12 h; and then diluted with MeOH and purified by preparative
reverse phase HPLC.
[0379] Method 4:
[0380] A mixture of an acid intermediate, such as 74, and of
1,1-carbonyldiimidazole in anhydrous THF could be heated to reflux
under nitrogen. After 2.5 h, amine was added and heating continued.
After an additional period of 3.about.20 h at reflux, the reaction
mixture could be cooled and concentrated in vacuo. The residue
could be purified by chromatography on silica gel to provide a
compound of Formula I.
[0381] In addition, the carboxylic acid could be converted to an
acid chloride using reagents such as thionyl chloride (neat or in
an inert solvent) or oxalyl chloride in a solvent such as benzene,
toluene, THF, or CH.sub.2Cl.sub.2. The amides could alternatively,
be formed by reaction of the acid chloride with an excess of
ammonia, primary, or secondary amine in an inert solvent such as
benzene, toluene, THF, or CH.sub.2Cl.sub.2 or with stoichiometric
amounts of amines in the presence of a tertiary amine such as
triethylamine or a base such as pyridine or 2,6-lutidine.
Alternatively, the acid chloride could be reacted with an amine
under basic conditions (usually sodium or potassium hydroxide) in
solvent mixtures containing water and possibly a miscible co
solvent such as dioxane or THF. Additionally, the carboxylic acid
could be converted to an ester preferably a methyl or ethyl ester
and then reacted with an amine. The ester could be formed by
reaction with diazomethane or alternatively trimethylsilyl
diazomethane using standard conditions which are well known in the
art. References and procedures for using these or other ester
forming reactions can be found in reference 52 or 54.
[0382] Additional references for the formation of amides from acids
are: Norman, M. H.; Navas, F. III; Thompson, J. B.; Rigdon, G. C.;
J. Med. Chem. 1996, 39(24), 4692-4703; Hong, F.; Pang, Y.-P.;
Cusack, B.; Richelson, E.; J. Chem. Soc., Perkin Trans 1 1997, 14,
2083-2088; Langry, K. C.; Org. Prep. Proc. Int. 1994, 26(4),
429-438; Romero, D. L.; Morge, R. A.; Biles, C.; Berrios-Pena, N.;
May, P. D.; Palmer, J. R.; Johnson, P. D.; Smith, H. W.; Busso, M.;
Tan, C.-K.; Voorman, R. L.; Reusser, F.; Althaus, I. W.; Downey, K.
M.; et al.; J. Med. Chem. 1994, 37(7), 999-1014; Bhattacharjee, A.;
Mukhopadhyay, R.; Bhattacharjya, A.; Indian J. Chem., Sect B 1994,
33(7), 679-682.
[0383] It is well known in the art that heterocycles may be
prepared from an aldehyde, carboxylic acid, carboxylic acid ester,
carboxylic acid amide, carboxylic acid halide, or cyano moiety or
attached to another carbon substituted by a bromide or other
leaving group such as a triflate, mesylate, chloride, iodide, or
phosponate. The methods for preparing such intermediates from
intermediates typified by the carboxylic acid intermediate, 69,
bromo intermediate, 76, or aldehyde intermediate, 70 described
above are known by a typical chemist practitioner. The methods or
types of heterocycles which may be constructed are described in the
chemical literature. Some representative references for finding
such heterocycles and their construction are included in reference
55 through 67 but should in no way be construed as limiting.
However, examination of these references shows that many versatile
methods are available for synthesizing diversely substituted
heterocycles and it is apparent to one skilled in the art that
these can be applied to prepare compounds of Formula I. Chemists
well versed in the art can now easily, quickly, and routinely find
numerous reactions for preparing heterocycles, amides, oximes or
other substituents from the above mentioned starting materials by
searching for reactions or preparations using a conventional
electronic database such as Scifinder (American Chemical Society),
Crossfire (Beilstein), Theilheimer, or Reaccs (MDS). The reaction
conditions identified by such a search can then be employed using
the substrates described in this application to produce all of the
compounds envisioned and covered by this invention. In the case of
amides, commercially available amines can be used in the synthesis.
Alternatively, the above mentioned search programs can be used to
locate literature preparations of known amines or procedures to
synthesize new amines. These procedures are then carried out by one
with typical skill in the art to provide the compounds of Formula I
for use as antiviral agents.
[0384] As shown below in Scheme 32, step a13, suitable substituted
azaindoles, such as the bromoazaindole intermediate, 76, may
undergo metal mediated couplings with aryl groups, heterocycles, or
vinyl stannanes to provide compounds of Formula I wherein R.sub.5
is aryl, heteroaryl, or heteroalicyclic for example. The
bromoazaindole intermediates, 76 (or azaindole triflates or
iodides) may undergo Stille-type coupling with heteroarylstannanes
as shown in Scheme 32, step a13. Conditions for this reaction are
well known in the art and references 68-70 as well as reference 52
provide numerous conditions in addition to the specific examples
provided in Scheme 33 and in the specific embodiments. It can be
well recognized that an indole stannane could also couple to a
heterocyclic or aryl halide or triflate to construct compounds of
Formula I. Suzuki coupling (reference 71) between the bromo
intermediate, 76, and a suitable boronate could also be employed
and some specific examples are contained in this application. 65 66
67
[0385] As shown in Scheme 34, step a14, aldehyde intermediates, 70,
could be used to generate numerous compounds of Formula I. The
aldehyde group could be a precursor for any of the substituents
R.sub.1 through R.sub.5 but the transormation for R.sub.5 is
depicted above for simplicity. The aldehyde intermediate 70, could
be reacted to become incorporated into a ring as described in the
claims or be converted into an acyclic group. The aldehyde, 70,
could be reacted with a Tosmic based reagent to generate oxazoles
(references 42 and 43 for example). The aldehyde, 70, could be
reacted with a Tosmic reagent and than an amine to give imidazoles
as in reference 72 or the aldehyde intermediate, 70, could be
reacted with hydroxylamine to give an oxime which is a compound of
Formula I as described below. Oxidation of the oxime with NBS,
t-butyl hypochlorite, or the other known reagents should provide
the N-oxide which react with alkynes or 3 alkoxy vinyl esters to
give isoxazoles of varying substitution. Reaction of the aldehyde
intermediate 70, with the known reagent, 77 (reference 70) shown
below under basic conditions would provide 4-aminotrityl oxazoles.
68
[0386] Removal of the trityl group should provide 4-amino oxazoles
which could be substitutued by acylation, reductive alkylation or
alkylation reactions or heterocycle forming reactions. The trityl
could be replaced with an alternate protecting group such as a
monomethoxy trityl, CBZ, benzyl, or appropriate silyl group if
desired. Reference 73 demonstrates the preparation of oxazoles
containing a triflouoromethyl moiety and the conditions described
therein demonstrates the synthesis of oxazoles with fluorinated
methyl groups appended to them.
[0387] The aldehyde could also be reacted with a metal or Grignard
(alkyl, aryl, or heteroaryl) to generate secondary alcohols. These
would be efficacious or could be oxidized to the ketone with TPAP
or MnO.sub.2 or PCC for example to provide ketones of Formula I
which could be utilized for treatment or reacted with metal
reagents to give tertiary alcohols or alternatively converted to
oximes by reaction with hydroxylamine hydrochlorides in ethanolic
solvents. Alternatively the aldehyde could be converted to benzyl
amines via reductive amination. An example of oxazole formation via
a Tosmic reagent is shown below in Scheme 35. The same reaction
would work with aldehydes at other positions and also in the 5 and
6 aza indole series. 69
[0388] Scheme 36 shows in step a15, a cyano intermediate, such as
62, which could be directly converted to compounds of Formula I via
heterocycle formation or reaction with organometallic reagents.
70
[0389] Scheme 37 shows a method for acylation of a cyanoindole
intermediate of formula 65' with oxalyl chloride which would give
acid chloride, 79', which could then be coupled with the
appropriate amine in the presence of base to provide 80'. 71
[0390] The nitrile intermediate, 80, could be converted to the
tetrazole of formula 81, which could then be alkylated with
trimethylsilyldiazometh- ane to give the compound of formula 82
(Scheme 38). 72
[0391] As shown in Scheme 38A, the nitrile intermediate 80 could be
derivatived to triazole of formula 80A by direct fusion with
hydrazides. Intermediate 80 could also be converted to the imidate
80B (or to thioaceamide), which could then be fused with hydrazides
to provide the triazole 80A. Alternatively, the acid intermediate
74 could be converted to the hydrazide 80C, which could then be
fused with thioacetamides to give triazole 80A. 73
[0392] Tetrazole alkylation with alkyl halides would be carried out
prior to azaindole acylation as shown in Scheme 39. Intermediate 65
could be converted to tetrazole, 83, which could be alkylated to
provide 84. Intermediate 84 could then be acylated and hydrolyzed
to provide 85 which could be subjected to amide formation
conditions to provide 86. The group appended to the tetrazole may
be quite diverse and still exhibit impressive potency. 74
[0393] Scheme 40 shows that an oxadiazole such as, 88, may be
prepared by the addition of hydroxylamine to the nitrile, 80,
followed by ring closure of intermediate 87 with phosgene.
Alkylation of oxadiazole, 88, with trimethylsilyldiazomethane would
give the compound of formula 89. 75
[0394] A 7-cyanoindole, such as 80, could be efficiently converted
to the imidate ester under conventional Pinner conditions using
1,4-dioxane as the solvent. The imidate ester can be reacted with
nitrogen, oxygen and sulfur nucleophiles to provide C7-substituted
indoles, for example: imidazolines, benzimidazoles,
azabenzimidazoles, oxazolines, oxadiazoles, thiazolines, triazoles,
pyrimidines and amidines etc. For example the imidate may be
reacted with acetyl hydrazide with heating in a nonparticipating
solvent such as dioxane, THF, or benzene for example. (aqueous base
or aqueous base in an alcoholic solvent may need to be added to
effect final dehydrative cyclization in some cases) to form a
methyl triazine. Other hydrazines can be used. Triazines can also
be installed via coupling of stannyl triazines with 4,5,6, or
7-bromo or chloro azaindoles. The examples give an example of the
formation of many of these heterocycles.
REFERENCES
[0395] (1) Das, B. P.; Boykin, D. W. J. Med. Chem. 1977, 20,
531.
[0396] (2) Czarny, A.; Wilson, W. D.; Boykin, D. W. J. Heterocyclic
Chem. 1996, 33, 1393.
[0397] (3) Francesconi, I.; Wilson, W. D.; Tanious, F. A.; Hall, J.
E.; Bender, B. C.; Tidwell, R. R.; McCurdy, D.; Boykin, D. W. J.
Med. Chem. 1999, 42, 2260.
[0398] Scheme 41 shows addition of either hydroxylamine or
hydroxylamine acetic acid to aldehyde intermediate 90 could provide
oximes of Formula 91. 76
[0399] An acid may be a precursor for substituents R.sub.1 through
R.sub.5 when it occupies the corresponding position such as R.sub.5
as shown in Scheme 42. 77 78
[0400] An acid intermediate, such as 69, could be used as a
versatile precursor to generate numerous substituted compounds. The
acid could be converted to hydrazonyl bromide and then a pyrazole
via reference 74. One method for general heterocycle synthesis
would be to convert the acid to an alpha bromo ketone (ref 75) by
conversion to the acid chloride using standard methods, reaction
with diazomethane, and finally reaction with HBr. The alpha bromo
ketone could be used to prepare many different compounds of Formula
I as it can be converted to many heterocycles or other compounds of
Formula I. Alpha amino ketones can be prepared by displacement of
the bromide with amines. Alternatively, the alpha bromo ketone
could be used to prepare heterocycles not available directly from
the aldeheyde or acid. For example, using the conditions of Hulton
in reference 76 to react with the alpha bromo ketone would provide
oxazoles. Reaction of the alpha bromoketone with urea via the
methods of reference 77 would provide 2-amino oxazoles. The alpha
bromoketone could also be used to generate furans using beta keto
esters (ref 78-80) or other methods, pyrroles (from beta
dicarbonyls as in ref 81 or by Hantsch methods (ref 82) thiazoles,
isoxazoles and imidazoles (ref 83) example using literature
procedures. Coupling of the aforementioned acid chloride with
N-methyl-O-methyl hydroxylamine would provide a "Weinreb Amide"
which could be used to react with alkyl lithiums or Grignard
reagents to generate ketones. Reaction of the Weinreb anion with a
dianion of a hydroxylamine would generate isoxazoles (ref 84).
Reaction with an acetylenic lithium or other carbanion would
generate alkynyl indole ketones, a transformation depicted in
Scheme 41a. Reaction of this alkynyl intermediate with diazomethane
or other diazo compounds would give pyrazoles (ref 85, Scheme 41a).
Reaction with azide or hydroxylamine would give heterocycles after
elimination of water. Nitrile oxides would react with the alkynyl
ketone to give isoxazoles (ref 86). Reaction of the initial acid to
provide an acid chloride using for example oxalyl chloride or
thionyl chloride or triphenyl phosphine/carbon tetrachloride
provides a useful intermediate as noted above. Reaction of the acid
chloride with an alpha ester substituted isocyanide and base would
give 2-substituted oxazoles (ref 87). These could be converted to
amines, alcohols, or halides using standard reductions or
Hoffman/Curtius type rearrangements.
[0401] Scheme 43 describes alternate chemistry for installing the
oxoacetyl piperazine moiety onto the 3 position of the azaindoles.
StepA'" in Scheme 43 depicts reaction with formaldehyde and
dimethylamine using the conditions in Frydman, B.; Despuy, M. E.;
Rapoport, H.; J. Am. Chem. Soc. 1965, 87, 3530 will provide the
dimethylamino compound shown.
[0402] Step B'" shows displacement with potassium cyanide would
provide the cyano derivative according to the method described in
Miyashita, K.; Kondoh, K.; Tsuchiya, K.; Miyabe, H.; Imanishi, T.;
Chem. Pharm. Bull. 1997, 45(5), 932-935 or in Kawase, M.;
Sinhababu, A. K.; Borchardt, R. T.; Chem. Pharm. Bull. 1990,
38(11), 2939-2946. The same transformation could also be carried
out using TMSCN and a tetrabutylammonium flouride source as in
Iwao, M.; Motoi, O.; Tetrahedron Lett. 1995, 36(33), 5929-5932.
Sodium cyanide could also be utilized. 79
[0403] Step C'" of Scheme 43 depicts hydrolysis of the nitrile with
sodium hydroxide and methanol would provide the acid via the
methods described in Iwao, M.; Motoi, O.; Tetrahedron Lett. 1995,
36(33), 5929-5932 for example. Other basic hydrolysis conditions
using either NaOH or KOH as described in Thesing, J.; et al.; Chem.
Ber. 1955, 88, 1295 and Geissman, T. A.; Armen, A.; J. Am. Chem.
Soc. 1952, 74, 3916. The use of a nitrilase enzyme to achieve the
same transformation is described by Klempier N, de Raadt A, Griengl
H, Heinisch G, J. Heterocycl. Chem., 1992 29, 93, and may be
applicable.
[0404] Step D'" of Scheme 43 depicts an alpha hydroxylation which
may be accomplished by methods as described in Hanessian, S.; Wang,
W.; Gai, Y.; Tetrahedron Lett. 1996, 37(42), 7477-7480; Robinson,
R. A.; Clark, J. S.; Holmes, A. B.; J. Am. Chem. Soc. 1993,115(22),
10400-10401 (KN(TMS).sub.2 and then camphorsulfonyloxaziridine or
another oxaziridine; and Davis, F. A.; Reddy, R. T.; Reddy, R. E.;
J. Org. Chem. 1992, 57(24), 6387-6389.
[0405] Step E'" of Scheme 43 shows methods for the oxidation of the
alpha hydroxy ester to the ketone which may be accomplished
according to the methods described in Mohand, S. A.; Levina, A.;
Muzart, J.; Synth. Comm. 1995, 25 (14), 2051-2059. A preferred
method for step E'" is that of Ma, Z.; Bobbitt, J. M.; J. Org.
Chem. 1991, 56(21), 6110-6114 which utilizes 4-(NH-Ac)-TEMPO in a
solvent such as CH.sub.2Cl.sub.2 in the presence of para
toluenesulfonic acid. The method described in Corson, B. B.; Dodge,
R. A.; Harris, S. A.; Hazen, R. K.; Org. Synth. 1941, I, 241 for
the oxidation of the alpha hydroxy ester to the ketone uses
KmnO.sub.4 as oxidant. Other methods for the oxidation of the alpha
hydroxy ester to the ketone include those described in Hunaeus,;
Zincke,; Ber. Dtsch Chem. Ges. 1877, 10, 1489; Acree,; Am. Chem.
1913, 50, 391; and Claisen,; Ber. Dtsch. Chem. Ges. 1877, 10,
846.
[0406] Step F'" of Scheme 43 depicts the coupling reactions which
may be carried out as described previously in the application and
by a preferred method which is described in Li, H.; Jiang, X.; Ye,
Y.-H.; Fan, C.; Romoff, T.; Goodman, M. Organic Lett., 1999, 1,
91-93 and employs
3-(Diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT); a
new coupling reagent with remarkable resistance to racemization.
80
[0407] Scheme 44 depicts the preparation of Formula I compounds by
coupling HWC(O)A to the acid as described in Step F'" of Scheme 43,
followed by hydroxylation as in Step D'" of Scheme 43 and oxidation
as described in Step E'" of Scheme 43. 81
[0408] Scheme 45 depicts a method for the preparation which could
be used to obtain amido compounds of Formula I. Step G' represents
ester hydrolysis followed by amide formation (Step H' as described
in Step F'" of Scheme 43). Step I' of Scheme 45 depicts the
preparation of the N-oxide which could be accomplished according to
the procedures in Suzuki, H.; Iwata, C.; Sakurai, K.; Tokumoto, K.;
Takahashi, H.; Hanada, M.; Yokoyama, Y.; Murakami, Y.; Tetrahedron
1997, 53(5), 1593-1606; Suzuki, H.; Yokoyama, Y.; Miyagi, C.;
Murakami, Y.; Chem. Pharm. Bull. 1991, 39(8), 2170-2172; and
Ohmato, T.; Koike, K.; Sakamoto, Y.; Chem. Pharm. Bull. 1981, 29,
390. Cyanation of the N-oxide is shown in Step J' of Scheme 45
which may be accomplished according to Suzuki, H.; Iwata, C.;
Sakurai, K.; Tokumoto, K.; Takahashi, H.; Hanada, M.; Yokoyama, Y.;
Murakami, Y.; Tetrahedron 1997, 53(5), 1593-1606 and Suzuki, H.;
Yokoyama, Y.; Miyagi, C.; Murakami, Y.; Chem. Pharm. Bull. 1991,
39(8), 2170-2172. Hydrolysis of the nitrile to the acid is depicted
in Step K' of Scheme 45 according to procedures such as Shiotani,
S.; Tanigucchi, K.; J. Heterocycl. Chem. 1996, 33(4), 1051-1056;
Memoli, K. A.; Tetrahedron Lett. 1996, 37(21), 3617-3618;
Adolfsson, H.; Waernmark, K.; Moberg, C.; J. Org. Chem. 1994,
59(8), 2004-2009; and El Hadri, A.; Leclerc, G.; J. Heterocycl.
Chem. 1993, 30(3), 631-635. Step L' of Scheme 45 depicts a method
which could be utilized for the preparation of amido compounds of
Formula I from the cyano derivative which may be accomplished
according to procedures described in Shiotani, S.; Taniguchi, K.;
J. Heterocycl. Chem. 1997, 34(2), 493-499; Boogaard, A. T.; Pandit,
U. K.; Koomen, G.-J.; Tetrahedron 1994, 50(8), 2551-2560; Rivalle,
C.; Bisagni, E.; Heterocycles 1994, 38(2), 391-397; and Macor, J.
E.; Post, R.; Ryan, K.; J. Heterocycl. Chem. 1992, 29(6),
1465-1467. Step M' of Scheme 45 shows a method which could be used
for the preparation of amido compounds of Formula I from the acid
derivative which may be accomplished according to procedures
described in Norman, M. H.; Navas, F. III; Thompson, J. B.; Rigdon,
G. C.; J. Med. Chem. 1996, 39(24), 4692-4703; Hong, F.; Pang,
Y.-P.; Cusack, B.; Richelson, E.; J. Chem. Soc., Perkin Trans 1
1997, 14, 2083-2088; Langry, K. C.; Org. Prep. Proced. Int. 1994,
26(4), 429-438; Romero, D. L.; Morge, R. A.; Biles, C.;
Berrios-Pena, N.; May, P. D.; Palmer, J. R.; Johnson, P. D.; Smith,
H. W.; Busso, M.; Tan, C.-K.; Voorman, R. L.; Reusser, F.; Althaus,
I. W.; Downey, K. M.; et al.; J. Med. Chem. 1994, 37(7), 999-1014
and Bhattacharjee, A.; Mukhopadhyay, R.; Bhattacharjya, A.; Indian
J. Chem., Sect B 1994, 33(7), 679-682. 82
[0409] Scheme 46 shows a method which could be used for the
synthesis of an azaindole acetic acid derivative. Protection of the
amine group could be effected by treatment with
di-tert-butyldicarbonate to introduce the t-Butoxycarbonyl (BOC)
group. Introduction of the oxalate moiety may then be accomplished
as shown in Step A of Scheme 46 according to the procedures
described in Hewawasam, P.; Meanwell, N. A.; Tetrahedron Lett.
1994, 35(40), 7303-7306 (using t-Buli, or s-buli, THF); or
Stanetty, P.; Koller, H.; Mihovilovic, M.; J. Org. Chem. 1992,
57(25), 6833-6837 (using t-Buli). The intermediate thus formed
could then be cyclized to form the azaindole as shown in Step B of
Scheme 46 according to the procedures described in Fuerstner, A.;
Ernst, A.; Krause, H.; Ptock, A.; Tetrahedron 1996, 52(21),
7329-7344 (using. TiCl3, Zn, DME); or Fuerstner, A.; Hupperts, A.;
J. Am. Chem. Soc. 1995, 117(16), 4468-4475 (using Zn, excess
Tms-Cl, TiCl3 (cat.), MeCN).
[0410] Scheme 49 provides another route to azaindole intermediates
which could then be further elaborated to provide compounds of
Formula I, such as the amido derivatives shown. Steps G" and H" of
Scheme 49 could be carried out according to the procedures
described in Takahashi, K.; Shibasaki, K.; Ogura, K.; Iida, H.;
Chem. Lett. 1983, 859; and Itoh, N.; Chem. Pharm. Bull. 1962, 10,
55. Elaboration of the intermediate to the amido compound of
Formula I could be accomplished as previously described for Steps
I'-M' of Scheme 45. 83
[0411] Scheme 50 shows the preparation of azaindole oxalic acid
derivatives. The starting materials in Scheme 50 could be prepared
according to Tetrahedron Lett. 1995, 36, 2389-2392. Steps A', B',
C', and D' of Scheme 50 may be carried out according to procedures
described in Jones, R. A.; Pastor, J.; Siro, J.; Voro, T. N.;
Tetrahedron 1997, 53(2), 479-486; and Singh, S. K.; Dekhane, M.; Le
Hyaric, M.; Potier, P.; Dodd, R. H.; Heterocycles 1997, 44(1),
379-391. Step E' of Scheme 50 could be carried out according to the
procedures described in Suzuki, H.; Iwata, C.; Sakurai, K.;
Tokumoto, K.; Takahashi, H.; Hanada, M.; Yokoyama, Y.; Murakami,
Y.; Tetrahedron 1997, 53(5), 1593-1606; Suzuki, H.; Yokoyama, Y.;
Miyagi, C.; Murakami, Y.; Chem. Pharm. Bull. 1991, 39(8),
2170-2172; Hagen, T. J.; Narayanan, K.; Names, J.; Cook, J. M.; J.
Org. Chem. 1989, 54, 2170; Murakami, Y.; Yokoyama, Y.; Watanabe,
T.; Aoki, C.; et al.; Heterocycles 1987, 26, 875; and Hagen, T. J.;
Cook, J. M.; Tetrahedron Lett. 1988, 29(20), 2421. Step F' of
Scheme 50 shows the conversion of the phenol to a fluoro, chloro or
bromo derivative. Conversion of the phenol to the fluoro derivative
could be carried out according to procedures described in Christe,
K. O.; Pavlath, A. E.; J. Org. Chem. 1965, 30, 3170; Murakami, Y.;
Aoyama, Y.; Nakanishi, S.; Chem. Lett. 1976, 857; Christe, K. O.;
Pavlath, A. E.; J. Org. Chem. 1965, 30, 4104; and Christe, K. O.;
Pavlath, A. E.; J. Org. Chem. 1966, 31, 559. Conversion of the
phenol to the chloro derivative could be carried out according to
procedures described in Wright, S. W.; Org. Prep. Proc. Int. 1997,
29(1), 128-131; Hartmann, H.; Schulze, M.; Guenther, R.; Dyes Pigm
1991, 16(2), 119-136; Bay, E.; Bak, D. A.; Timony, P. E.;
Leone-Bay, A.; J. Org. Chem. 1990, 55, 3415; Hoffmann, H.; et al.;
Chem. Ber. 1962, 95, 523; and Vanallan, J. A.; Reynolds, G. A.; J.
Org. Chem. 1963, 28, 1022. Conversion of the phenol to the bromo
derivative could be carried out according to procedures described
in Katritzky, A. R.; Li, J.; Stevens, C. V.; Ager, D. J.; Org.
Prep. Proc. Int. 1994, 26(4), 439-444; Judice, J. K.; Keipert, S.
J.; Cram, D. J.; J. Chem. Soc., Chem. Commun. 1993, 17, 1323-1325;
Schaeffer, J. P.; Higgins, J.; J. Org. Chem. 1967, 32, 1607; Wiley,
G. A.; Hershkowitz, R. L.; Rein, R. M.; Chung, B. C.; J. Am. Chem.
Soc. 1964, 86, 964; and Tayaka, H.; Akutagawa, S.; Noyori, R.; Org.
Syn. 1988, 67, 20. 84
[0412] Scheme 51 describes methods for the preparation of azaindole
acetic acid derivatives by the same methods employed for the
preparation of azaindole oxalic acid derivatives as shown and
described in Scheme 50 above. The starting material employed in
Scheme 51 could be prepared according to J. Org. Chem. 1999, 64,
7788-7801. Steps A", B", C", D", and E" of Scheme 51 could be
carried out in the same fashion as previously described for Steps
Steps A', B', C', D', and E' of Scheme 50. 85
[0413] As shown in Scheme 52, the pieces HW-A can be prepared by a
number of different methods. One useful way is by reacting a mono
protected piperazine with a heteroaryl chloride, bromide, iodide,
or triflate. This reaction is typically carried out at elevated
temperature (50 to 250 degrees celsius) in a solvent such as
ethylene glycol, DME, dioxane, NMP, or DMF. A tertiary amine base
such as triethyl amide or diisopropyl ethyl amine is typically
employed and usually 2 to 4 equivalents are employed. At least 2
equivalents are used if a salt of HWA is utilized. The piperazine
is typically monoprotected with a BOC group since this material is
commercially available. Removal of the Boc group is typically done
using HCl (typically 1 to 6N) in dioxane to provide the HCl salt.
TFA may also be used to generate the TFA salt. Alternatively, the
conditions for coupling heterocycles using copper catalysis
discussed earlier in Scheme 12 may be used to couple W to A via
displacement of X in X-A. Alternatively Palladium catalysis in the
presence of a bidentate catalyst via the procedures of Buckwald or
the use of a ferrocenyl catalyst via the methods of Hartwig could
be used to couple the piperazine to the heteroaryl (A).
[0414] The preparations of the naphthyridine (X-A) starting
materials have been previously disclosed in the following
references:
[0415] (1) Rapoport, H.; Batcho, A. D. J. Org. Chem. 1963, 28,
1753.
[0416] (2) Baldwin, J. J.; Mensler, K.; Ponticello, G. S. J. Org.
Chem. 1978, 43, 4878.
[0417] (3) Baldwin, J. J.; Mensler, K.; Ponticello, G. S. U.S. Pat.
No. 4,176,183. 86 87
[0418] Scheme 53 describes how a protected piperazine can be
coupled to Q-COOH via standard methodology in described in step D
of Schemes A and 1a-1e. Conditions for removal of the amine
protecting group which could be tBoc or other groups is protecting
group specific. As shown in Scheme 53 where tBoc is the preferred
protecting group used to exemplify the strategy, standard
conditions for removal such as TFA in dichloromethane or
alternatively aqueous HCl can provide the free amine. The free
amine is coupled to A using the conditions described in Scheme 52
for step F"".
[0419] Chemistry
[0420] All .sup.1H NMR spectra were recorded on a 500 MHz Bucker
DRX-500f instrument, unless otherwise (e.g. 300 MHz Bucker DPX-300)
stated. All Liquid Chromatography (LC) data were recorded on a
Shimadzu LC-10AS liquid chromatograph using a SPD-10AV UV-Vis
detector with Mass Spectrometry (MS) data determined using a
Micromass Platform for LC in electrospray mode.
[0421] LC/MS Method (i.e., Compound Identification)
[0422] Note: column A is used unless otherwise indicated in the
preparation of intermediates or examples.
2 Column A: YMC ODS-A S7 3.0 .times. 50 mm column Column B:
PHX-LUNA C18 4.6 .times. 30 mm column Column C: XTERRA ms C18 4.6
.times. 30 mm column Column D: YMC ODS-A C18 4.6 .times. 30 mm
column Column E: YMC ODS-A C18 4.6 .times. 33 mm column Column F:
YMC C18 S5 4.6 .times. 50 mm column Column G: XTERRA C18 S7 3.0
.times. 50 mm column Gradient: 100% Solvent A/0% Solvent B to 0%
Solvent A/100% Solvent B R.sub.t in min. Gradient time: 2 minutes
Hold time 1 minute Flow rate: 5 mL/min Detector Wavelength: 220 nm
Solvent A: 10% MeOH/90% H.sub.2O/0.1% Trifluoroacetic Acid Solvent
B: 10% H.sub.2O/90% MeOH/0.1% Trifluoroacetic Acid
[0423] Compounds purified by preparative HPLC were diluted in MeOH
and purified using the following methods on a Shimadzu LC-10A
automated preparative HPLC system or on a Shimadzu LC-8A automated
preparative HPLC system with detector (SPD-10AV UV-VIS) wavelength
and solvent systems (A and B) the same as above.
[0424] Preparative HPLC Method (i.e., Compound Purification)
[0425] 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)
3 Solvent A: 10% MeOH/90% H.sub.2O/0.1% Trifluoroacetic Acid
Solvent B: 10% H.sub.2O/90% MeOH/0.1% Trifluoroacetic Acid Column:
YMC C18 S5 20 .times. 100 mm column Detector Wavelength: 220 nm
[0426] General and Example Procedures Excerpted from Analogous
Oxoacetyl Piperazineamide Applications
[0427] The procedures described references 93-95 and 106 are
applicable example procedures for synthesizing the compounds of
formula I in this application and the intermediates used for their
synthesis. The following guidelines are illustrative but not
limiting.
[0428] The general Bartoli (vinyl Magnesium bromide) methods for
preparing functionalized indoles or azaindoles dexcribed in the
applications can be utilized for preparing new indoles or
azaindoles from the appropriate nitro aromatics or heteroaromatics
for this application. For example, in PCT/US02/00455, the general
procedure for preparing intermediate 2a (7-chloro-6-azaindole) from
2-chloro-3-nitro pyridine can be considered a general procedure
illustrating conditions which can be used to prepare azaindoles for
this application. Similarly, the general procedure from the same
application to prepare intermediate 3a, Methyl
(7-chloro-6azaindol-3-yl) oxoacetate, provides experimental details
for carrying our Step B of (Schemes 1-7 in this application).
Similarly, the general procedure from the same application to
prepare intermediate 4a (Potassium(7-chloro-6azaindol-3-yl)
oxoacetate, provides an example of the general method for
hydrolying oxoacteic esters (Step C of Schemes 1-1c, 3-7). General
procedures for carrying out the same steps in the indole series are
provided in references 93 and 95. An example Bartoli reaction
preparation of a functionalized indole is given in the preparation
of intermediate 1 of PCT/US01/20300 where the preparation of
4-fluoro-7-bromo-azaindole is described from
2-fluoro-5-bromonitrobenzene- . The following Scheme provides an
example of the preparation of 4,7-dibromo-6-azaindole via an
extension of this methodology. 88
[0429] Subsequent procedures for the preparation of intermediates 2
and 3 describe procedures for adding the alkyl oxoacetate and then
for ester hydrolysis to provide the carboxylate salt and then the
carboxylic acid after acidification. Thus the chemistry described
in the incoprorated previous applications for preparing azaindole
and indole intermediates is applicable since the desired compounds
are the same.
[0430] Procedures for carrying out the coupling of the indole or
azaindole oxoacetic acids to piperazine amides are described in the
references 93-95 and 106. These can also be used as procedures for
preparing the N-heteroaryl piperazines of this invention by taking
the experimental procedures and substituting a N-heteroaryl
piperazine or mono protected piperazine in place of the piperazine
amide. This is possible because both groups have a free amine with
relatively similar activity and since the other portions of both
the piperazine benzamide and the N-heteroaryl piperazine are
relatively unreactive to many conditions, they can be installed
similarly. For example, the preparation of intermediate 4 of
PCTJUS01/20300 and the preparation of intermediate 5a of
PCT/US02/00455 describe couplings of a piperazine benzamide or
methyl piperazine benzamide to an indole or azaindole oxoacetic
acid or carboxylate salt respectively. (The acid or salt can be
used interchangeably). These same procedures can be used directly
for the preparation of the compounds of this invention by
substituting the desired N-heteroaryl piperazines for the
piperazine amides utilized in earlier applications.
[0431] Preparation of Intermediate 5a from PCT/US02/00455 89
[0432] can be used as a procedure for 90
[0433] Preparation of Intermediate 4 from PCT/US01/20300 91
[0434] can be used as a procedure for 92
[0435] Once attached via a similar amide bond, both the piperazine
benzamides and the N-heteroaryl piperazines moieties are relatively
inert and thus reaction conditions used for functionalizing indoles
or azaindoles in the presence of piperazine benzamides are useful
for carrying out the same tranformations in the presence of the
N-heteroaryl piperazines. Thus the methods and transformations
described in references 93-95 and 106 including the experimental
procedures which describe methods to functionalize the indole or
azaindole moiety in the piperazine amide series are generally
applicable for construction and functionalization of the
N-heteroaryl piperazines of this invention. These same applications
describe general methods and specific preparations for obtaining
stannane and boronic acid reagents used for synthesizing the
compounds of Formula I.
[0436] Preparation of Example 1 from PCT/US02/00455
[0437] Typical Boron/palladium coupling procedure 93
[0438] can be used as a procedure for 94
[0439] or even as a procedure for 95
[0440] where R.sup.x is as described for Scheme 6B
[0441] Preparation of Example 39 from PCT/US02/00455
[0442] An example of the typical stannane/palladium coupling
procedure 96
[0443] can be used as a procedure for 97
[0444] or even as a procedure for 98
[0445] where R.sup.x is as described for Scheme 6B
[0446] Preparation of Example 20 from PCT/US01/20300
[0447] An example to show how functionalization procedures of
oxoacetyl piperazine benzamides can be used to carry out similar
tranformations in the corresponding piperidine alkenes 99
[0448] can be used as a procedure for 100
[0449] or even as a procedure for 101
[0450] where R.sup.x is as described for Scheme 6B
[0451] Preparation of Intermediates and Examples:
[0452] All starting materials, unless otherwise indicated can be
purchased from commercial sources. Methods are given for the
preparation of intermediates.
[0453] Note: Unless Otherwise indicated, HPLC conditions utilized
column G. 102
[0454] To a mixture of 1-chloroisoquinoline (527 mg, 3.22 mmol) and
tert-butyl 1-piperazinecarboxylate (500 mg, 2.68 mmol) in ethylene
glycol (8 ml) at r.t. was added triethylamine (2.0 ml, 14.3 mmol).
The reaction mixture was then stirred at 100.degree. C. for 6 to 20
h. After cooling to r.t., the mixture was diluted with water (30
ml), basified using saturated aqueous NaHCO.sub.3, and extracted
with CH.sub.2Cl.sub.2 (50 ml). The organic extract was evaporated
in vacuo and the residue purified by flash column chromatography
(0% to 10% EtOAc/hexane) to give Intermediate 1 as a white solid.
.sup.1H NMR: (300 MHz, CD.sub.3OD) .delta. 8.19 (d, 1H, J=8.4),
8.06 (d, 1H, J=5.7), 7.84 (d, 1H, J=8.1), 7.69 (b t, 1H), 7.60 (b
t, 1H), 7.38 (d, 1H, J=5.7), 3.71-3.69 (b s, 4H), 3.33-3.30 (b s,
4H), 1.50 (s, 9H); LC/MS: (ES+) m/z (M+H).sup.+=314; HPLC
R.sub.t=1.063.
[0455] A mixture of Intermediate 1 (40 mg, 0.128 mmol) in a
solution of HCl in 1,4-dioxane (0.5 ml, 4 N) was stirred at r.t.
for 3 h. The excess reagent and volatile were then evaporated, and
the residue further dried under high vacuum to give the
hydrochloride salt of Intermediate 2 as a white solid. .sup.1H NMR:
(300 MHz, CD.sub.3OD) .delta. 8.39 (d, 1H, J=8.7), 8.15-8.05
(overlapping m, 2H), 7.98-7.89 (overlapping m, 2H), 7.77 (d, 1H,
J=6.6), 4.11-4.08 (m, 4H), 3.67-3.64 (m, 4H); LC/MS: (ES+) m/z
(M+H).sup.+=214; HPLC R.sub.t=0.207. 103
[0456] To a mixture of 4-fluoro-7-cyanoindole (1.0 g, 6.24 mmol) in
EtOH (50 ml) was added hydroxylamine hydrochloride (651 mg, 9.37
mmol) and triethylamine (1.7 ml). The reaction mixture was refluxed
for 16 hours. After removal of the volatile under high vacuum, the
residue was added water (10 ml) and filtered to afford the crude
hydroxyamidine intermediate. To this intermediate was added
triethylorthoformate (10 ml) and the mixture heated at 110.degree.
C. for 16 hours. After removal of most of the excess reagent, the
residue was purified by flash chromatography with
(CH.sub.2Cl.sub.2) to give intermediate 2aa as pale yellow solid
(419 mg, 33%). .sup.1H NMR (CDCl.sub.3) .delta. 9.90 (s, 1H), 8.80
(s, 1H), 8.01 (app dd, J=8.3, 4.8, 1H), 7.34 (app t, J=2.8, 1H),
6.93 (app dd, J=9.8, 8.3, 1H), 6/74 (app dd, J=3.2, 2.3, 1H); LC/MS
(ES+) m/z (M+H).sup.+=204, HPLC R.sub.t=1.910, Column YMC ODS-A C18
S7 (3.0.times.50 mm), Gradient Time=2 min, Flow rate 5 ml/min.
104
[0457] To a solution of intermediate 2aa (200 mg, 0.984) in
CH.sub.2Cl.sub.2 (10 ml) was added oxalyl chloride (1 ml), and the
reaction mixture stirred under gentle reflux for 16 hours. Removal
of solvent in vacuo and the excess reagent under high vacuum
afforded intermediate 4aa as a yellow solid, which was used without
further purification.
[0458] The following HPLC conditions for the LCMS were used for
compounds 2ac, 3aa, 2ad, 3ab, 4ab, and 4ac: Column: Xterra C18 S7
3.times.50 mm; Gradient Time=3 min; Flow rate=4 ml/min. 105
[0459] To a mixture of 2ab (2.0 g, 7.3 mmol) and CuCN (1.0 g, 11
mmol) was added DMF (20 ml). The reaction mixture was heated at
150.degree. C. for 1 hour. After cooling to room temperature, the
reaction mixture was added NaOMe (20 ml, 25 wt. % solution in
MeOH), and was heated at 110.degree. C. for 10 minutes. After
cooling to room temperature, the reaction mixture was poured into
an aqueous solution of ammonium acetate (sat. 500 ml). The
resulting mixture was filtered through a short Celite.RTM. pad. The
filtrate was extracted with EtOAc (4.times.500 ml). The combined
extracts were dried over MgSO.sub.4 and evaporated in vacuo to give
a brownish residue, which was triturated with MeOH (5 ml.times.3)
to provide 2ac as a yellow solid (317 mg, 25%). The structure was
supported by NOE experiments. .sup.1H NMR: (DMSO-d.sub.6) 12.47 (s,
1H), 8.03 (s, 1H), 7.65 (t, J=2.8, 1H), 6.70 (dd, J=2.8, 1.8, 1H),
4.08 (s, 3H); LC/MS: (ES+) m/z (M+H).sup.+=174; HPLC
R.sub.t=1.320.
[0460] Preparation of Compound 3aa:
[0461] To 1-ethyl-3-methylimidazolium chloride (85 mg, 0.58 mmol)
in a capped vial was quickly added aluminum chloride (231 mg, 1.73
mmol). The mixture was vigorously stirred at room temperature until
the formation of the ionic liquid. After cooling to room
temperature, the ionic liquid was added compound 2ac (50 mg, 0.29
mmol) and ethyl chlorooxoacetate (0.2 ml, 1.79 mmol). The reaction
mixture was stirred at room temperature for three hours, cooled to
0.degree. C. and quenched by carefully adding ice-water (15 ml).
The precipitates were filtered, washed with water (3.times.5 ml)
and dried in vacuo to give 3aa as a grayish yellow solid (50 mg,
63%). .sup.1H NMR: (DMSO-d.sub.6) 13.73 (s, 1H), 8.54 (s, 1H), 8.26
(s, 1H), 4.35 (q, J=7.0, 2H), 4.06 (s, 3H), 1.29 (t, J=7.0, 3H);
LC/MS: (ES+) m/z (M+H).sup.+=274; HPLC R.sub.t=1.527.
[0462] Preparation of Compound 4ab:
[0463] To a mixture of 3aa (200 mg, 0.73 mmol) in MeOH (1 ml) was
added NaOH (2.5 ml, 1N aqueous). The reaction mixture was stirred
at room temperature for 30 minutes, and then acidified with
hydrochloric acid (3 ml, 1N) to pH about 2. The solid was filtered,
washed with water (4.times.5 ml), and dried in vacuo to give 4ab as
a brownish solid (160 mg, 89%). Compound 4ab was used without
further purification. LC/MS: (ES+) m/z (M+H).sup.+=246; HPLC
R.sub.t=0.777. 106
[0464] A mixture of 2ab, 4,7-dibromo-6-azaindole (2.0 g, 7.0 mmol),
CuBr (2.0 g, 14 mmol) and NaOMe (20 ml, 25 wt. % solution in MeOH)
was heated in a sealed tube at 100.degree. C. for 12h. Aftering
cooling to r.t., the mixture was diluted with MeOH (20 ml) and then
filtered. The filtrate was purified by preparative reverse phase
HPLC using the method: Start % B=0, Final % B=50, Gradient time=10
min, Flow Rate=45 mL/min, Column: Xterra MS C18 5 um 30.times.50
mm, Fraction Collection: 2.20-4.30 min. LC/MS: (ES+) m/z
(M+H).sup.+=179, HPLC R.sub.t=0.857.
[0465] Compound 3ab was prepared in a similar manner to compound
3aa.
[0466] Intermediate 4ac was prepared in a similar manner to
Intermediate 4ab. LC/MS: (ES+) m/z (M+H).sup.+=251, HPLC
R.sub.t=0.503.
[0467] Intermediate 4ad 107
[0468] To 4-methoxy-7-bromoindole (500 mg, 2.21 mmol) was added a
solution of oxalyl chloride in CH.sub.2Cl.sub.2 (10 ml, 20 mmol, 2
M), and the mixture was stirred at r.t. for 16h. The solvent and
the excess reagent were then evaporated and the crude product used
for the next step without further purification. 108
[0469] A mixture of acid chloride intermediate 4aa (37 mg, 0.126
mmol) and Intermediate 2 (0.128 mmol) in CH.sub.2Cl.sub.2 (1 ml) at
r.t. was added N,N-diisopropylethylamine (0.18 ml, 1.03 mmol), and
the reaction mixture stirred for 17 h. The mixture was evaporated
to dryness and the volatile further removed under high vacuum. The
solid residue was then treated with water (3 ml), filtered, and
further washed with water (3.times.2 ml) and minimum amount of MeOH
(2.times.1 ml) to obtain Example 1 as a white solid. .sup.1H NMR:
(500 MHz, CDCl.sub.3) .delta. 10.59 (s, 1H), 8.85 (s, 1H), 8.22 (d,
1H, J=3.0), 8.18-8.10 (overlapping m, 3H), 7.79 (b d, 1H), 7.68 (b
m, 1H), 7.60 (b m, 1H), 7.32 (d, 1H, J=5.5), 7.14 (dd, 1H, J=8.5,
10.0), 4.06 (b m, 2H) 3.86 (b m, 2H), 3.64-3.42 (b m, 4H); LC/MS:
(ES+) m/z (M+H).sup.+=471; HPLC R.sub.t=1.210.
[0470] Intermediate 4, Intermediate 5 and Example 2 were prepared
in a manner analogous to the methods used for Example 1. 109
[0471] Intermediate 4: LC/MS: (ES+) m/z (M+H).sup.+=304; HPLC
R.sub.t=1.053.
[0472] Intermediate 5: LC/MS: (ES+) m/z (M+H).sup.+=204; HPLC
R.sub.t=0.083.
[0473] Example 2: .sup.1H NMR: (500 MHz, CD.sub.3OD) .delta. 9.40
(s, 1H), 8.25 (s, 1H), 8.16 (dd, 1H, J=4.5, 8.0), 7.88 (s, 1H),
7.87 (d, 1H, J=5.5), 7.18 (dd, 1H, J=8.0, 10.3), 7.10 (d, 1H,
J=5.5), 6.86 (s, 1H), 3.97 (m, 2H), 3.94 (m, 2H) 3.85 (m, 2H), 3.70
(m, 2H); LC/MS: (ES+) m/z (M+H).sup.+=461; HPLC R.sub.t=1.073.
[0474] Intermediate 6, Intermediate 7 and Example 3 were prepared
in the same manner as described for Example 1. 110
[0475] Intermediate 6: LC/MS: (ES+) m/z (M+H).sup.+=315; HPLC
R.sub.t=0.968.
[0476] Intermediate 7: Hydrochloride salt .sup.1H NMR: (CD.sub.3OD)
.delta. 8.84 (s, 1H), 8.29 (d, J=10, 1H), 8.11 (app t, J=10, 1H),
7.90 (d, J=10, 1H), 7.83 (app t, J=10, 1H), 4.53 (b s, 4H), 3.53 (b
s, 4H).
[0477] LC/MS: (ES+) m/z (M+H).sup.+=215; HPLC R.sub.t=0.080.
[0478] Example 3: .sup.1H NMR: (500 MHz, DMSO-d.sub.6) .delta.
12.30 (s, 1H), 9.87 (s, 1H), 8.68 (s, 1H), 8.21 (b d, 1H),
8.10-8.06 (overlapping m, 2H), 7.84 (b d, 2H), 7.57 (m, 1H), 7.28
(app t, 1H), 3.88 (s, 4H), 3.74 (b s, 2H), 3.64 (b s, 2H); LC/MS:
(ES+) m/z (M+H).sup.+=472; HPLC R.sub.t=1.000.
[0479] LCMS Conditions:
[0480] Solvent A: 10% MeOH--90% H2O--0.1% TFA
[0481] Solvent B: 90% MeOH--10% H2O--0.1% TFA
[0482] Column: XTERRA C18 S7 3.0.times.50 mm
[0483] Start % B=0
[0484] Final % B=100
[0485] Gradient Time=2 min
[0486] Flow Rate=5 ml/min
[0487] Wavelength=220 111
EXAMPLE 4
[0488] LC/MS: (ES+) m/z (M+H).sup.+=493, 495; HPLC R.sub.t=1.128.
112
[0489] A mixture of Example 4 (50 mg, 0.101 mmol), imidazole (69
mg, 1.01 mmol) cesium carbonate (66 mg, 0.203 mmol) and copper
bromide (30 mg, 0.212 mmol) was heated at 145.degree. C. for 4 h.
The reaction mixture was then cooled to r.t., diluted with MeOH (2
ml) and filtered. The residue was further washed with 3.times.2 ml
MeOH. The filtrate was evaporated in vacuo to give the crude
product, which was purified by preparative TLC (10%
MeOH/CH.sub.2Cl.sub.2) to give Example 5; LC/MS: (ES+) m/z
(M+H).sup.+=481; HPLC R.sub.t=0.867. 113
EXAMPLE 6
[0490] Example 6 was prepared in the same manner as Example 5.
[0491] LC/MS: (ES+) m/z (M+H).sup.+=480, 495; HPLC
R.sub.t=1.233.
[0492] The following HPLC conditions for the LCMS were used for
Example 7, Example 8 and Example 9: Column: G; Gradient Time=3 min;
Flow rate=4 ml/min. 114
[0493] Preparation of Example 7:
[0494] To a mixture of 4ab (crude, about 1.94 mmol), DEPBT (1.161
g, 3.88 mmol), intermediate 2 (952 mg, 2.91 mmol) in DMF (5 ml) was
added N,N-diisopropylethylamine (3.0 ml, 17 mmol). The reaction
mixture was stirred at room temperature for 16 hours. The reaction
mixture was then diluted with MeOH (6 ml) and filtered. The
filtrate was purified by preparative reverse phase HPLC using the
method: Start % B=20, Final % B=60, Gradient time=15 min, Flow
Rate=40 ml/min, Column: XTERRA C18 5 .mu.m 30.times.50 mm, Fraction
Collection: 6.169-6.762 min. .sup.1H NMR: (DMSO-d.sub.6) 13.71 (s,
1H), 8.50 (d, J=3.0, 1H), 8.27 (s, 1H), 8.23 (d, J=8.5, 1H), 8.06
(d, J=6.0, 1H), 7.97 (d, J=8.0, 1H), 7.82 (app t, J=7.5, 1H), 7.69
(d, J=7.5, 1H), 7.51 (d, J=6.0, 1H), 4.16 (s, 3H), 3.93 (b s,2),
3.66 (b s, 2H), 3.63 (b s, 2H), 3.48 (b s, 2H); LC/MS: (ES+) m/z
(M+H).sup.+=441, HPLC R.sub.t=1.200.
[0495] Preparation of Example 8:
[0496] Anhydrous hydrogen chloride gas was bubbled through a
suspension of Example 7 (160 mg, 2.18 mmol) in MeOH (5 ml) at
0.degree. C. for 15 minutes. After evaporation of most of the
volatile, the residue was purified by preparative reverse phase
HPLC using the method: Start % B=20, Final % B=60, Gradient time=15
min, Flow Rate=40 ml/min, Column: XTERRA C18 5 .mu.m 30.times.50
mm, Fraction Collection: 6.169-6.762 min. .sup.1H NMR:
(DMSO-d.sub.6) 12.49 (s, 1H), 8.25 (m, 1H), 8.17 (s, 1H), 8.11 (d,
J=9.5, 1H), 8.04 (m, 1H), 7.99 (d, J=8.5, 1H), 7.70 (app t, J=7.5,
1H), 7.53 (d, J=6.5, 1H), 7.0 (b s, 2H), 4.08 (s, 3H), 3.93 (b s,
2), 3.66 (b s, 4H), 3.50 (b s, 2H); LC/MS: (ES+) m/z
(M+H).sup.+=459, HPLC R.sub.t=1.237. 115
[0497] Preparation of Example 9:
[0498] To a mixture of Intermediate 4ac (crude, about 0.56 mmol),
DEPBT (336 mg, 1.12 mmol) and intermediate 7 (220 mg, 0.67 mmol) in
DMF (3 ml) was added N,N-diisopropylethylamine (1.0 ml, 5.7 mmol).
The reaction mixture was stirred at room temperature for 16 hours.
The reaction mixture was then diluted with MeOH (4 ml), and
filtered. The filtrate was purified by preparative reverse phase
HPLC using the method: Start % B=0, Final % B=55, Gradient time=15
min, Flow Rate=40 ml/min, Column: XTERRA C18 5 .mu.m 30.times.100
mm, Fraction Collection: 8.71-9.16 min. .sup.1H NMR: (DMSO-d.sub.6)
13.07 (s, 1H), 8.91 (s, 1H), 8.23 (m, 2H), 8.05 (app t, J=7.5, 1H),
7.85 (d, J=8.0, 1H), 7.73 (app t, J=7.7, 1H), 7.48 (s,1H), 4.36 (b
s, 2H), 4.20 (b s, 2H), 4.00 (s, 3H), 3.85 (s, 3H), 3.63 (b s, 4H);
LC/MS: (ES+) m/z (M+H).sup.+=447, HPLC R.sub.t=0.987.
EXAMPLE 43
[0499] 116
[0500] A mixture of intermediate 4ab (0.671 g, 2.7 mmol),
intermediate 2 (0.869 g, 4.1 mmol), EDC (0.928 g, 4.8 mmol),
dimethylaminopyridine (0.618 g, 5.1 mmol) and N-methylmorpholine
(2.4 ml, 21.6 mmol) in DMF (20 ml) was stirred at room temperature
for 17 hr. The reaction mixture was then quenched with 1N HCl and
extracted with ethyl acetate (6 times). The combined organic
extracts were evaporated in vacuo and purified by flash
columatography (0%.fwdarw.5% MeOH/CH.sub.2Cl.sub.2) to provide
Example 43 as a dark solid; .sup.1H NMR (CDCl.sub.3) .delta. 9.64
(b s, 1H), 8.15 (d, J=5.5, 1H), 8.11 (d, J=8.0, 1H), 8.05 (d,
J=3.0, 1H), 7.79 (d, J=8.0, 1H), 7.65 (app t, J=9.0, 1H), 7.57 (d,
J=8.5, 2H), 7.32 (d, J=6.0, 1H), 6.74 (d, J=8.5, 1H), 4.05 (s, 3H,
overlapping with m), 4.05-4.00 (m, 2H), 3.79 (b s, 2H), 3.56 (b s,
2H), 3.48 (b s, 2H); LC/MS (ES+) m/z (M+H).sup.+=440, HPLC
R.sub.t=0.993. 117
EXAMPLE 10
[0501] Example 10 was prepared by refluxing Example 43 (30 mg,
0.068 mmol) in 5N NaOH (0.8 ml, 40 mmol) for 17 hr. The reaction
mixture was acidified by adding 1N HCl, and extracted with ethyl
acetate (3 times). The crude product was purified by preparative
reverse phase HPLC to give a brown film; Separation method: Start %
B=0, Final % B=100, Gradient time=6 min, Flow Rate=30 ml/min,
Column: YMC C18 S5 20.times.50 mm; .sup.1H NMR: (CD.sub.3OD)
.delta. 8.40 (d, J=8.5, 1H), 8.19 (s, 1H), 8.06-7.98 (m, 2H),
7.86-7.811 (m, 3H), 7.61 (d, J=8.0, 1H), 6.86 (d, J=8.5, 1H), 4.12
(b s, 2H), 4.03 (s, 3H), 3.95 (b s, 2H), 3.86 (d, J=4, 4H); LC/MS:
(ES+) m/z (M+H).sup.+=458, HPLC R.sub.t=0.787. 118
[0502] A solution of Example 43 (49 mg, 0.11 mmol) in anhydrous
EtOH (0.5 ml, 200 proof) in a re-usable sealed tube was bubbled
with anhydrous HCl gas at room temperature for approximately 15
min. After which time, the tube was closed and the mixture stirred
at room temperature for 72 hours. The volatiles were then
evaporated, and intermediate 8 was used without further
purification; LC/MS: (ES+) m/z (M+H).sup.+=486, HPLC R.sub.t=0.837.
119
EXAMPLE 11
[0503] To a mixture of intermediate 8 (53 mg, 0.11 mmol) in
anhydrous EtOH (1.5 ml, 200 proof) was added acetic hydrazide (45
mg, 0.61 mmol) and N,N-diisopropylethylamine (0.1 ml, 0.57 mmol).
The reaction mixture was heated to 150.degree. C. and refluxed for
3 hours. After cooling to room temperature, sodium methoxide (24
mg, 0.44 mmol) was added and the mixture was refluxed for 2
additional hours. The reaction was quenched with 1N HCl and diluted
with H.sub.2O. The crude product was purified by preparative
reverse phase HPLC using the separation method: Start % B=25, Final
% B=65, Gradient time=20 min, Flow Rate=30 ml/min, Column: YMC C18
S5 20.times.50 mm; .sup.1H NMR (CD.sub.3OD) .delta. 8.41 (d, J=8.5,
1H), 8.23 (s, 1H), 8.23-8.01 (m, 2H), 7.90 (d, J=8.0, 1H),
7.84-7.82 (m, 2H), 7.64 (d, J=6.5, 1H), 6.94 (d, J=8.5, 1H),
4.14-4.12 (m, 2H), 4.03 (s, 3H), 4.01-3.99 (m, 2H), 3.89 (s, 4H),
2.62 (s, 3H); LC/MS (ES+) m/z (M+H).sup.+=496, HPLC R.sub.t=0.927.
120
EXAMPLE 12
[0504] Example 12 was obtained as a side product from the reaction
to make Example 11 and was isolated via preparative reverse phase
HPLC using the same method as above; .sup.1H NMR (CD.sub.3OD)
.delta. 8.41 (d, J=8.5, 1H), 8.18 (s, 1H), 8.07-7.99 (m, 2H),
7.86-7.79 (m, 3H), 7.64 (d, J=6.5, 1H), 6.86 (d, J=8.5, 1H), 4.12
(b s, 2H), 4.03 (s, 3H), 3.99 (b s, 2H), 3.87 (s, 4H); LC/MS (ES+)
m/z (M+H).sup.+=458, HPLC R=0.807. 121
EXAMPLE 13
[0505] Example 13 was also obtained as a side-product from the
reaction to make Example 11. The compound was isolated via
preparative reverse phase HPLC using the same method as above;
.sup.1H NMR (CD.sub.3OD) .delta. 8.38 (d, J=8.0, 1H), 8.18 (s, 1H),
8.03-7.94 (m, 3H), 7.90 (d, J=6.5, 1H), 7.82 (app t, J=7.8, 1H),
7.60 (d, J=6.0, 1H), 6.91 (d, J=9.0, 1H), 4.47 (q, J=7.0, 2H),
4.11-4.10 (m, 2H), 4.05 (s, 3H), 3.90-3.85 (m, 4H), 3.78-3.74 (m,
2H), 1.44 (t, J=7.0, 3H); LC/MS: (ES+) m/z (M+H).sup.+=487, HPLC
R.sub.t=1.170. 122
EXAMPLE 14
[0506] A mixture of hydroxylamine hydrochloride (20 mg, 0.29 mmol)
and triethylamine (50 .mu.l, 0.36 mmol) in anhydrous EtOH (1.5 ml,
200 proof) was added to Example 43 (80 mg, 0.18 mmol), and the
resulting mixture stirred at room temperature. After 48 hours, the
mixture was added additional hydroxylamine hydrochloride (47 mg,
0.68 mmol) and triethylamine (80 .mu.l, 0.58 mmol), and stirred for
5 days. The precipitates were filtered and washed with excess EtOH
to give Example 14 as a white solid. Further purification was
performed by reverse phase preparative HPLC using the method: Start
% B=0, Final % B=100, Gradient time=7 min, Flow Rate=30 ml/min,
Column: YMC C18 S5 20.times.50 mm; .sup.1H NMR: (CD.sub.3OD)
.delta. 8.38 (d, J=9.0, 1H), 8.21 (s, 1H), 8.03 (d, J=7.0, 1H),
7.96-7.91 (m, 2H), 7.81 (app t, J=7.8, 1H), 7.60 (d, J=6.5, 1H),
7.53 (d, J=8.5, 1H), 6.97 (d, J=8.5, 1H), 4.11-4.10 (m, 2H), 4.05
(s, 3H), 3.87 (s, 4H), 3.79 (s, 2H); LC/MS: (ES+) m/z
(M+H).sup.+=473, HPLC R.sub.t=0.663. 123
EXAMPLE 15
[0507] To a mixture of Example 43 (80 mg, 0.18 mmol) in DMF (2 ml)
was added sodium azide (35 mg, 0.54 mmol) and ammonium chloride (29
mg, 0.54 mmol). The resulting mixture was heated to 90.degree. C.
and allowed to stir for 20 hr. After cooling to 0.degree. C. in an
ice-water bath, the reaction was quenched by adding several drops
of 1N HCl and then diluted with water, upon which a precipitate was
formed. The solids were filtered and washed with an excess of water
to give Example 15 as a white solid; .sup.1H NMR: (CD.sub.3OD)
.delta. 8.25-8.22 (m, 2H), 8.07 (d, J=5.5, 1H), 7.91 (dd, J=8.0,
8.0, 2H), 7.72 (app t, J=7.5, 1H), 7.63 (app t, J=8.0, 1H), 7.42
(d, J=6.0, 1H), 7.00 (d, J=8.0, 1H), 4.06 (s, 3H, overlapping with
m), 4.06-4.03 (m, 2H), 3.79 (b s, 2H), 3.55 (b s, 2H), 3.43 (b s,
2H); LC/MS: (ES+) m/z (M+H).sup.+=483, HPLC R.sub.t=0.947. 124
EXAMPLE 16
[0508] To a mixture of Example 15 (35 mg, 0.073 mmol) in MeOH (0.5
mL)/PhH (0.8 mL) at room temperature was added
trimethylsilyldiazomethane (80 .mu.l, 0.16 mmol, 2M in hexanes).
After stirring for 2.5 hours, the reaction mixture was cooled to
0.degree. C. in an ice-water bath and quenched using excess acetic
acid. The volatiles were evaporated in vacuo, and the residue
purified by reverse phase preparative HPLC using the method: Start
% B=20, Final % B=60, Gradient time=20 min, Flow Rate=30 ml/min,
Column: YMC C18 S5 20.times.50 mm, Fraction Collection: 8.37-8.97
min; .sup.1H NMR: (CD.sub.3OD) .delta. 8.40 (d, J=8.5, 1H), 8.25
(s, 1H), 8.10 (d, J=8.5, 1H), 8.05 (d, J=8.0, 1H), 7.99 (app t,
J=7.3, 1H), 7.88 (d, J=6.5, 1H), 7.83 (app t, J=7.8, 1H), 7.61 (d,
J=6.5, 1H), 6.98 (d, J=8.5, 1H), 4.48 (s, 3H), 4.13 (b s, 2H), 4.06
(s, 3H), 3.93 (b s, 2H), 3.89 (b s, 2H), 3.83 (b s, 2H). The
position of N-methyl group was supported by HMBC NMR studies;
LC/MS: (ES+) m/z (M+H).sup.+=497, HPLC R.sub.t=1.083. 125
EXAMPLE 17
[0509] A flask charged with Example 14 (45 mg, 0.095 mmol) was
added triethyl orthoformate (1 ml, 6.0 mmol) and the mixture heated
at 110.degree. C. for 24 hours. The volatiles were evaporated and
the residue subjected to purification by reverse phase preparative
HPLC using the method: Start % B=20, Final % B=60, Gradient time=20
min, Flow Rate=30 mL/min, Column: YMC C18 S5 20.times.50 mm,
Fraction Collection: 7.73-8.10 min; .sup.1H NMR: (CD.sub.3OD)
.delta. 9.34 (s, 1H), 8.39 (d, J=8.5, 1H), 8.24 (s, 1H), 8.13 (d,
J=8.5, 1H), 8.04 (d, J=8.0, 1H), 7.98 (app t, J=7.8, 1H), 7.89 (d,
J=6.5, 1H), 7.83 (app t, J=7.8, 1H), 7.61 (d, J=6.5, 1H), 7.00 (d,
J=8.5, 1H), 4.14-4.12 (m, 2H), 4.06 (s, 3H), 3.92-3.87 (m, 4H),
3.82-3.80 (m, 2H); LC/MS: (ES+) m/z (M+H).sup.+=483, HPLC
R.sub.t=1.057. 126
EXAMPLE 18
[0510] To a solution of intermediate 8 (30 mg, 0.062 mmol) in
anhydrous EtOH (1.0 ml, 200 proof) was added cyclopropylamine (50
.mu.L, 0.67 mmol). The reaction mixture was stirred at room
temperature for 8 hours, then diluted with MeOH and subjected to
purification via reverse phase preparative HPLC using the method:
Start % B=0, Final % B=100, Gradient time=8 min, Flow Rate=30
mL/min, Column: YMC C18 S5 20.times.50 mm, Fraction Collection:
3.13-3.59 min. .sup.1H NMR: (CD.sub.3OD) .delta. 8.41 (d, J=8.5,
1H), 8.22 (s, 1H), 8.06-7.98 (m, 2H), 7.89-7.82 (m, 2H), 7.63 (d,
J=6.0, 1H), 7.55 (d, J=8.0, 1H), 6.96 (d, J=8.5, 1H), 4.12 (b s,
2H), 4.04 (s, 3H), 3.95 (b s, 2H), 3.77 (b s, 4H), 2.86 (b s, 1H),
1.08 (b s, 2H), 0.93 (b s, 2H); LC/MS: (ES+) m/z (M+H).sup.+=497,
HPLC R.sub.t=0.720. 127
EXAMPLE 19
[0511] To a solution of intermediate 8 (30 mg, 0.062 mmol), in
anhydrous EtOH (1.0 ml, 200 proof) was added 1,2-phenylenediamine
(30 mg, 0.27 mmol). The mixture was stirred at room temperature for
8 hours, then diluted with MeOH and subjected to purification by
reverse phase preparative HPLC using the method: Start % B=0, Final
% B=100, Gradient time=8 min, Flow Rate=30 ml/min, Column: YMC C18
S5 20.times.50 mm, Fraction Collection: 4.24-4.82 min; .sup.1H NMR:
(CD.sub.3OD) .delta. 8.40 (d, J=8.5, 1H), 8.31 (s, 1H), 8.04-7.78
(m, 7H), 7.61 (d, J=6.5, 1H), 7.52 (b s, 2H), 7.08 (d, J=8.0, 1H),
4.14-4.09 (m, 2H), 4.09 (s, 3H, overlapping with m), 3.92-3.84 (m,
6H); LC/MS: (ES+) m/z (M+H).sup.+=531, HPLC R.sub.t=0.957. 128
EXAMPLE 20
[0512] To a solution of Example 14 (45 mg, 0.095 mmol) in pyridine
(1.0 ml) was added acetyl chloride (50 .mu.L, 0.70 mmol). The
reaction mixture was heated to 115.degree. C. for two hours and
then cooled to room temperature. After dilution with MeOH, the
crude mixture was purified by preparative reverse phase HPLC using
the method: Start % B=0, Final % B=100, Gradient time=16 min, Flow
Rate=30 mL/min, Column: YMC C18 S5 20.times.50 mm; .sup.1H NMR:
(CD.sub.3OD) .delta. 8.38 (d, J=8.0, 1H), 8.22 (s, 1H), 8.04-7.89
(m, 4H), 7.79 (s, 1H), 7.59 (d, J=6.0, 1H), 6.97 (d, J=8.0, 1H),
4.17-4.00 (m, 2H), 4.04 (s, 3H, overlapping with m), 3.87 (b s,
3H), 3.78 (b s, 3H), 2.70 (s, 3H); LC/MS: (ES+) m/z
(M+H).sup.+=497, HPLC R.sub.t=1.123.
EXAMPLE 21
[0513] 129
[0514] To intermediate 8 (40 mg, 0.082 mmol) in EtOH (2.0 mL) was
added formyl hydrazide (25 mg, 0.41 mmol) and
N,N-diisopropylethylamine (50 .mu.L, 0.28 mmol). The mixture was
refluxed at 130.degree. C. for five hours. After cooling to room
temperature, the mixture was diluted with MeOH, and purified by
preparative reverse phase HPLC using the separation method: Start %
B=0, Final % B=100, Gradient time=9 min, Flow Rate=30 mL/min,
Column: YMC C18 S5 20.times.50 mm, Fraction Collection: 4.53-5.08
min. .sup.1H NMR: (CD.sub.3OD) .delta. 8.53 (s, 1H), 8.42 (d,
J=8.4, 1H), 8.25 (s, 1H), 8.07-8.01 (m, 2H), 7.98 (d, J=8.4, 1H),
7.87-7.81 (m, 2H), 7.64 (d, J=6.6, 1H), 6.95 (d, J=8.4, 1H),
4.15-4.12 (m, 2H), 4.03 (s, 3H), 4.00-3.97 (m, 2H), 3.88 (s, 4H);
LC/MS: (ES+) m/z (M+H).sup.+=482, HPLC R.sub.t=0.920.
EXAMPLE 22
[0515] 130
[0516] Example 22 was prepared in a similar manner as Example 21.
Purification of the desired product was performed by preparative
reverse phase HPLC using the separation method: Start % B=0, Final
% B=100, Gradient time=12 min, Flow Rate=30 mL/min, Column: YMC C18
S5 20.times.50 mm, Fraction Collection: 5.93-6.49 min. The fraction
collected was evaporated and further purified by using the method:
Start % B=15, Final % B=80, Gradient time=20 min, Flow Rate=30
mL/min, Column: Xterra Prep MS C18 5 um 19.times.50 mm, Fraction
Collection: 6.91-7.34 min. .sup.1H NMR: (CD.sub.3OD) .delta. 8.41
(d, J=8.4, 1H), 8.32 (s, 1H), 8.06-7.80 (m, 9H), 7.63 (d, J=6.6,
1H), 7.01 (d, J=8.4, 1H), 4.15-4.13 (m, 2H), 4.07 (s, 3H),
3.98-3.85 (m, 6H); LC/MS: (ES+) m/z (M+H).sup.+=559, HPLC
R.sub.t=1.023.
EXAMPLE 23
[0517] 131
[0518] Example 23 was isolated as an intermediate/side product in
the reaction to make Example 22. Purification was performed by
preparative reverse phase HPLC using the separation method: Start %
B=0, Final % B=100, Gradient time=12 min, Flow Rate=30 mL/min,
Column: YMC C18 S5 20.times.50 mm, Fraction Collection: 3.82-4.22
min. .sup.1H NMR: (CD.sub.3OD) .delta. 9.20 (d, J=2.1, 1H), 8.85
(dd, J=8.0, 2.5, 1H), 8.50 (d t, J=14.0, 3.0, 1H), 8.43 (d, J=8.4,
1H), 8.31 (s, 1H), 8.08-7.97 (m, 2H), 7.89-7.81 (m, 2H), 7.76 (d,
J=8.4, 1H), 7.71-7.66 (m, 1H), 7.65 (d, J=6.9, 1H), 7.06 (d, J=8.4,
1H), 4.15-4.13 (m, 2H), 4.09 (s, 3H), 4.00-3.90 (m, 6H); LC/MS:
(ES+) m/z (M+H).sup.+=577, HPLC R.sub.t=0.707
EXAMPLE 24
[0519] 132
[0520] Example 24 was prepared in a similar manner as Example 21.
Purification of the desired product was performed by preparative
reverse phase HPLC using the separation method: Start % B=0, Final
% B=100, Gradient time=12 min, Flow Rate=30 mL/min, Column: YMC C18
S5 20.times.50 mm, Fraction Collection: 5.57-6.14 min. .sup.1H NMR:
(CD.sub.3OD) .delta. 8.40 (d, J=8.4, 1H), 8.25 (s, 1H), 8.05-7.96
(m, 2H), 7.87 (d, J=6.9, 2H), 7.82-7.79 (m, 1H), 7.62 (d, J=6.6,
1H), 6.94 (d, J=8.7, 1H), 4.16 (s, 2H), 4.15-4.11 (m, 2H), 4.03 (s,
3H), 3.96-3.93 (m, 2H), 3.87-3.85 (m, 4H); LC/MS: (ES+) m/z
(M+H).sup.+=521, HPLC R.sub.t=0.983.
EXAMPLE 25
[0521] 133
[0522] Example 25 was isolated as an intermediate in the reaction
to make Example 24. Purification was performed by preparative
reverse phase HPLC using the separation method: Start % B=0, Final
% B=100, Gradient time=12 min, Flow Rate=30 mL/min, Column: YMC C18
S5 20.times.50 mm, Fraction Collection: 4.67-5.28 min. .sup.1H NMR:
(CD.sub.3OD) .delta. 8.40 (d, J=8.7, 1H), 8.18 (s, 1H), 8.05-7.93
(m, 2H), 7.88-7.79 (m, 3H), 7.60 (d, J=10.2, 1H), 6.87 (d, J=8.4,
1H), 4.13-4.10 (m, 2H), 4.03 (s, 3H), 3.94-3.90 (m, 2H), 3.85-3.82
(m, 6H); LC/MS: (ES+) m/z (M+H).sup.+=539, HPLC R.sub.t=0.797.
EXAMPLE 26
[0523] 134
[0524] Example 15 (96 mg, 0.20 mmol) was dissolved in acetonitrile
(1.5 mL), and to the mixture was added methyl bromoacetate (40
.mu.L, 0.42 mmol), followed by potassium carbonate (38 mg, 0.27
mmol). The mixture was stirred at room temperature for three hours
and the precipitate was then filtered to obtain the product, which
was pure by .sup.1HNMR and LC/MS analysis. The filtrate was
extracted with EtOAc (4 times) and the combined extracts evaporated
to obtain additional crude product. The crude product was purified
using preparative reverse phase HPLC with the separation method:
Start % B=0, Final % B=100, Gradient time=20 min, Flow Rate=20
mL/min, Column: YMC C18 S5 20.times.50 mm, Fraction Collection:
10.85-11.31 min. .sup.1H NMR: (CD.sub.3OD) .delta. 8.40 (d, J=8.5,
1H), 8.28 (s, 1H), 8.17 (d, J=8.5, 1H), 8.05 (d, J=8.0, 1H), 7.95
(d, J=6.5, 2H), 7.83 (t, J=7.5, 1H), 7.62 (t, J=6.5, 1H), 7.04 (d,
J=8.0, 1H), 5.80 (s, 2H), 4.15 (b s, 2H), 4.08 (s, 3H), 3.89-3.87
(m, 6H), 3.87 (s, 3H), 3.78 (b s, 2H); LC/MS: (ES+) m/z
(M+H).sup.+=555, HPLC R.sub.t=1.087. Alkylation at the tetrazole N2
was supported by HMBC NMR analysis.
EXAMPLE 27
[0525] 135
[0526] To a mixture of Example 26 (110 mg, 0.20 mmol) in MeOH (1.5
mL) at ambient temperature was added 1N NaOH (0.5 mL, 0.50 mmol)
and stirred for three hours. The reaction was then quenched with 1N
HCl (10 drops) to induce precipitation of the product. The
precipitates were filtered, washed with excess H.sub.2O, and dried
under high vacuum to give Example 27 as an off-white solid. .sup.1H
NMR: (CD.sub.3OD) .delta. 8.27 (d, J=8.5, 1H), 8.24 (s, 1H), 8.13
(d, J=8.5, 1H), 8.03 (d, J=6.0, 1H), 7.90 (d, J=8.5, 1H), 7.78 (t,
J=7.5, 1H), 7.68 (t, J=7.8, 1H), 7.46 (d, J=6.0, 1H), 6.99 (d,
J=8.5, 1H), 4.37 (b s, 2H), 4.12-4.04 (m, 2H), 4.05 (s, 3H),
3.82-3.80 (m, 2H), 3.64 (m, 2H), 3.53-3.51 (m, 2H); LC/MS: (ES+)
m/z (M+H).sup.+=541, HPLC R.sub.t=1.003.
EXAMPLE 28
[0527] 136
[0528] To a mixture of Example 27 (20 mg, 0.037 mmol) in DMF (1.5
mL) was added methylamine hydrochloride (12 mg, 0.39 mmol), HOBT
(28 mg, 0.21 mmol), EDC (40 mg, 0.21 mmol) and NMM (50 .mu.L, 0.45
mmol). The reaction mixture was stirred at ambient temperature for
twenty-four hours, diluted with MeOH and then purified by
preparative reverse phase HPLC using the separation method: Start %
B 0, Final % B=100, Gradient time=18 min, Flow Rate=30 mL/min,
Column: Xterra Prep MS C18 5 um 19.times.50 mm, Fraction
Collection: 7.43-7.88 min. .sup.1H NMR: (CD.sub.3OD) .delta. 8.39
(d, J=8.5, 1H), 8.26 (s, 1H), 8.13 (d, J=8.5, 1H), 8.04 (d, J=7.0,
1H), 7.98 (t, J=7.5, 1H), 7.88 (d, J=6.5, 1H), 7.83 (t, J=8.3, 1H),
7.61 (d, J=6.0, 1H), 7.00 (d, J=8.5, 1H), 5.53 (s, 2H), 4.14-4.12
(m, 2H), 4.05 (s, 3H), 3.93-3.87 (m, 4H), 3.82-3.80 (m, 2H), 2.83
(s, 3H); LC/MS: (ES+) m/z (M+H).sup.+=554, HPLC R.sub.t=0.953.
EXAMPLE 29
[0529] 137
[0530] To a mixture of Example 27 (20 mg, 0.037 mol) in DMF (1.5
mL) was added NH.sub.4Cl (16 mg, 0.30 mol), HOBT (35 mg, 0.26
mmol), EDC (42 mg, 0.22 mmol), and NMM (50 .mu.L, 0.45 mmol). The
reaction mixture was stirred at ambient temperature for twenty-four
hours, diluted with MeOH and then purified by preparative reverse
phase HPLC using the separation method: Start % B=0, Final % B=100,
Gradient time=18 min, Flow Rate=30 mL/min, Column: Xterra Prep MS
C18 5 um 19.times.50 mm, Fraction Collection: 7.11-7.51 min.
.sup.1H NMR: (CD.sub.3OD) .delta. 8.38 (d, J=8.5, 1H), 8.24 (s,
1H), 8.10 (d, J=8.5, 1H), 8.03 (d, J=8.0, 1H), 7.97 (t, J=7.5, 1H),
7.87 (d, J=6.5, 1H), 7.82 (t, J=7.8, 1H), 7.59 (d, J=6.5, 1H), 6.97
(d, J=8.0, 1H), 5.57 (s, 2H), 4.13-4.11 (m, 2H), 4.04 (s, 3H),
3.92-3.87 (m, 4H), 3.81-3.79 (m, 2H); LC/MS: (ES+) m/z
(M+H).sup.+=540, HPLC R.sub.t=0.910.
EXAMPLE 30
[0531] 138
[0532] To a mixture of Example 4 (31.5 mg, 63.8 mmol) and
2-tributylstannyl pyridazine (30 mg, 81.3 mmol) in 1,4-dioxane (4
ml) in a re-usable sealed tube at r.t. was added
Pd(PPh.sub.3).sub.4 (20 mg, 17.3 mmol). The tube was tightly
closed, and the mixture stirred at 135.degree. C. for 3 h. After
cooled to r.t., the mixture was diluted with MeOH (4 ml), filtered
through a cake of celite and the filtrate evaporated. The resulting
residue was titurated with hexane (3.times.2 ml), and the hexane
removed by pipet. The residue was dried under vacuum, dissolved in
MeOH and purified by preparative reverse phase HPLC using the
separation method: Start % B=0, Final % B=100, Gradient time=6 min,
Flow Rate=30 mL/min, Column: Xterra Prep MS C18 5 um 19.times.50
mm, Fraction Collection: 3.72-4.24 min. .sup.1H NMR: (CD.sub.3OD)
.delta. 9.30 (s, 1H), 8.73 (app t, 1H), 8.48 (d, J=3.0, 1H), 8.42
(d, J=9.0, 1H), 8.25 (s, 1H), 8.08-8.06 (m, 1H), 8.07 (d, J=8.5,
1H), 8.03-8.00 (m, 1H), 7.84 (d, J=6.5, 2H), 7.64 (d, J=6.5, 1H),
6.99 (d, J=8.5, 1H), 4.17-4.14 (m, 2H), 4.06 (s, 3H), 4.00-3.98 (m,
2H), 3.90 (b s, 4H); LC/MS: (ES+) m/z (M+H).sup.+=493, HPLC
R.sub.t=1.063.
EXAMPLE 31
[0533] 139
[0534] Example 31 was prepared in a similar manner as described
before and purified by preparative reverse-phase HPLC using the
separation method: Start % B=20, Final % B=80, Gradient time=6 min,
Flow Rate=30 mL/min, Column: Xterra Prep MS C18 5 .mu.m 19.times.50
mm, Fraction Collection: 2.35-2.96 min. .sup.1H NMR: (CD.sub.3OD)
.delta. 9.08 (s, 1H), 8.42 (d, J=8.0, 1H), 8.30 (s, 1H), 8.20 (s,
1H), 8.08-8.01 (m, 2H), 7.87-7.83 (m, 2H), 7.65-7.64 (m, 1H), 7.60
(d, J=8.5, 1H), 6.91 (d, J=8.5, 1H), 4.15-4.13 (m, 2H), 4.03 (s,
3H), 4.03-4.00 (m, overlapped with s, 3H), 3.91 (b s, 2H),
3.91-3.88 (m, overlapped with s, 1H); LC/MS: (ES+) m/z
(M+H).sup.+=482, HPLC R.sub.t=0.893.
EXAMPLE 32
[0535] 140
[0536] To a solution of Example 24 (45 mg, 0.086 mmol) in MeOH (1.0
mL) in a re-usable sealed tube was bubbled with anhydrous hydrogen
chloride gas for 15 min. The tube was closed, and the mixture
stirred at ambient temperature for 3 hour. The volatiles were
evaporated in vacuo to give Example 32. .sup.1H NMR: (CD.sub.3OD)
.delta. 8.44 (d, J=8.5, 1H), 8.26 (s, 1H), 8.10-8.03 (m, 2H), 7.93
(d, J=8.5, 1H), 7.89 (t, J=7.5, 1H), 7.83 (d, J=6.5, 1H), 7.67 (d,
J=7.0, 1H), 6.99 (d, J=8.0, 1H), 4.15 (b s, 3H), 4.05 (b s, 4H),
3.94 (b m, 4H), 3.81 (s, 3H), 3.34 (s, 2H); LC/MS: (ES+) m/z
(M+H).sup.+=554, HPLC R.sub.t=0.997.
EXAMPLE 33
[0537] 141
[0538] To a mixture of Example 32 (13 mg, 0.024 mmol) in MeOH (0.5
mL) was added 1 N NaOH (0.1 mL), and stirred for 2 hours at room
temperature. The reaction was then quenched with 1N HCl (0.1 mL),
and the volatiles evaporated to give a clear film. .sup.1H NMR:
(CD.sub.3OD) .delta. 8.44 (d, J=6.5, 1H), 8.26 (s, 1H), 8.11-7.99
(m, 2H), 7.90-7.83 (m, 3H), 7.65 (d, J=6.0, 1H), 7.01 (d, J=8.0,
1H), 4.27 (b s, 1H), 4.16 (b s, 2H), 4.07 (b s, 4H), 3.97 (b d,
4H), 3.34 (s, 2H); LC/MS: (ES+) m/z (M+H).sup.+=540, HPLC
R.sub.t=0.917.
EXAMPLE 34
[0539] 142
[0540] To a mixture of Example 33 (23 mg, 0.043 mmol) in DMF (1.5
mL) were added methylamine hydrochloride (10 mg, 0.32 mmol), HOBT
(31 mg, 0.23 mmol), EDC (43 mg, 0.22 mmol), and NMM (50 .mu.L, 0.45
mmol). The mixture was stirred overnight at room temperature, and
then kept in the freezer over 48 hours. The desired product was
isolated by preparative reverse-phase HPLC using the separation
method: Start % B=0, Final % B=100, Gradient time=18 min, Flow
Rate=30 mL/min, Column: Xterra Prep MS C18 5 .mu.m 19.times.50 mm,
Fraction Collection: 6.73-7.34 min. .sup.1H NMR: (CD.sub.3OD)
.delta. 8.40 (d, J=8.5, 1H), 8.23 (s, 1H), 8.06-8.00 (m, 2H), 7.91
(d, J=8.5, 1H), 7.86-7.81 (m, 2H), 7.63 (d, J=7.0, 1H), 6.92 (d,
J=8.5, 1H), 4.13 (b s, 2H), 4.02 (s, 3H), 4.00-3.98 (m, 2H), 3.88
(b s, 4H), 3.84 (s, 2H), 2.80 (s, 3H); LC/MS: (ES+) m/z
(M+H).sup.+=553, HPLC R.sub.t=0.900.
EXAMPLE 35
[0541] 143
[0542] To a mixture of Example 33 (23 mg, 0.043 mmol) in DMF (1.5
mL) was added ammonium chloride (15 mg, 0.28 mmol), HOBT (35 mg,
0.28 mmol), EDC (43 mg, 0.22 mmol), and NMM (50 .mu.L, 0.45 mmol).
The mixture was stirred overnight at room temperature, and then
kept in the freezer over 48 hours. The desired product was isolated
by preparative reverse-phase HPLC using the separation method:
Start % B=0, Final % B=100, Gradient time=18 min, Flow Rate=30
mL/min, Column: Xterra Prep MS C18 5 .mu.m 19.times.50 mm, Fraction
Collection: 6.18-6.78 min. .sup.1H NMR: (CD.sub.3OD) .delta. 8.42
(d, J=8.5, 1H), 8.25 (s, 1H), 8.07 (d, J=8.0, 1H), 8.04 (t, J=7.3,
1H), 7.93 (d, J=8.5, 1H), 7.87-7.82 (m, 2H), 7.64 (d, J=6.5, 1H),
6.93 (d, J=8.0, 1H), 4.14 (b s, 2H), 4.03 (s, 3H), 4.00 (b s, 2H),
3.89 (s, 4H), 3.88 (s, 2H); LC/MS: (ES+) m/z (M+H).sup.+=539, HPLC
R.sub.t=0.850.
EXAMPLE 36
[0543] 144
[0544] Intermediate 4ad N Example 36 To a mixture of Intermediate
4ad (22 mg, 0.077 mmol) in DMF (1 mL) was added piperazine
hydrochloride Intermediate 2 (85 mg, 0.40 mmol), DEPBT (72 mg, 0.24
mmol), and N,N-diisopropylethylamine (0.11 mL, 0.57 mmol). The
reaction mixture was stirred for 18 hours at room temperature, and
the desired product was isolated by preparative reverse phase HPLC
using the following method: Start % B=0, Final % B=60, Gradient
time=18 min, Flow Rate=30 mL/min, Column: Xterra Prep MS C18 5
.mu.m 19.times.50 mm, Fraction Collection: 9.53-10.14 min. .sup.1H
NMR: (CD.sub.3OD) .delta. 9.38 (s, 1H), 8.44 (d, J=8.5, 1H), 8.38
(s, 1H), 8.34 (s, 1H), 8.09 (d, J=8.0, 1H), 8.05 (t, J=7.5, 1H),
7.94 (s, 1H), 7.88-7.84 (m, 2H), 7.67 (d, J=7.0, 1H), 4.16 (b s,
2H), 4.12 (s, 3H), 4.03 (b s, 2H), 3.92 (s, 4H); LC/MS: (ES+) m/z
(M+H).sup.+=483, HPLC R.sub.t=0.930. 145
[0545] Typical procedure for preparing azaindole from
nitropyridine: Preparation of 7-chloro-6-azaindole, Precursor 2a,
is an example of Step A of Scheme 1. 2-chloro-3-nitropyridine
(5.0g, 31.5 mmol) was dissolved in dry THF (200 m]L). After the
solution was cooled to -78.degree. C., vinyl magnesium bromide
(1.0M in THF, 100 mL) was added dropwise. The reaction temperature
was maintained at -78.degree. C. for 1 h, and then at -20.degree.
C. for another 12 h before it was quenched by addition of 20%
NH.sub.4Cl aqueous solution (150 mL). The aqueous phase was
extracted with EtOAc (3.times.150 mL). The combined organic layer
was dried over MgSO.sub.4, filtered and the filtrate was
concentrated in vacuo to give a residue which was purified by
silica gel column chromatography (EtOAc/Hexane, 1/10) to afford
1.5g (31%) of 7-chloro-6-azaindole, Precursor 2a. .sup.1H NMR (500
MHz, CD.sub.3OD) .delta. 7.84 (d, 1H, J=10.7 Hz), 7.55 (dd, 1H,
J=10.9, 5.45 Hz), 6.62 (d, 1H, J=5.54 Hz), 4.89 (s, 1H). MS m/z:
(M+H).sup.+ calcd for C.sub.7H.sub.6ClN.sub.2: 153.02; found
152.93. HPLC retention time: 0.43 minutes (column A). 146
[0546] Typical procedure for acylation of azaindole: Preparation of
Methyl (7-chloro-6-azaindol-3-yl)-oxoacetate, Precursor 3a is an
example of Step B of Scheme 1. 7-Chloro-6-azaindole, Precursor 2a
(0.5 g, 3.3 mmol) was added to a suspension of AlCl.sub.3 (2.2 g,
16.3 mmol) in CH.sub.2Cl.sub.2 (100 mL). Stirring was continued at
rt for 10 minutes before methyl chlorooxoacetate (2.0 g, 16.3 mmol)
was added dropwise. The reaction was stirred for 8 h. The reaction
was quenched with iced aqueous NH.sub.4OAc solution (10%, 200 mL).
The aqueous phase was extracted with CH.sub.2Cl.sub.2 (3.times.100
mL). The combined organic layer was dried over MgSO.sub.4, filtered
and the filtrate was concentrated in vacuo to give a residue which
was carried to the next step without further purification.
Precursor 2, Methyl (7-chloro-6-azaindol-3-yl)-oxoacetate: MS m/z:
(M+H).sup.+ calcd for C.sub.10H.sub.8ClN.sub.2O.sub.3: 239.02;
found 238.97. HPLC retention time: 1.07 minutes (column A). 147
[0547] Typical procedure of hydrolysis of ester: Preparation of
Potassium (7-chloro-6-azaindol-3-yl)-oxoacetate, Precursor 4a, is
an example of Step C of Scheme 1. Crude methyl
(7-chloro-6-azaindol-3-yl)-oxoacetate, Precursor 3a, and an excess
of K.sub.2CO.sub.3 (2 g) were dissolved in MeOH (20 mL) and
H.sub.2O (20 mL). After 8 h, the solution was concentrated and the
residue was purified by silica gel column chromatography to provide
200 mg of Potassium (7-chloro-6-azaindol-3-yl)-- oxoacetate. MS
m/z: (M+H).sup.+ of the corresponding acid was observed. Calc'd for
C.sub.9H.sub.6ClN.sub.2O.sub.3: 225.01; found 225.05. HPLC
retention time: 0.83 minutes (column A). 148
[0548] Precursor 2g, 7-chloro-4-azaindole was prepared by the same
method as Precursor 2a, starting from 4-Chloro-3-nitro-pyridine
(HCl salt, available from Austin Chemical Company, Inc.). MS m/z:
(M+H).sup.+ calcd for C.sub.7H.sub.6ClN.sub.2: 153.02; found
152.90. HPLC retention time: 0.45 minutes (column A). 149
[0549] Precursor 3f, Methyl (7-chloro-4-azaindol-3-yl)-oxoacetate
was prepared by the same method as Precursor 3a, starting from
Precursor 2g, 7-chloro-4-azaindole. MS m/z: (M+H).sup.+ calcd for
C.sub.10H.sub.8ClN.sub.2O.sub.3: 239.02; found 238.97. HPLC
retention time: 0.60 minutes (column A). 150
[0550] Precursor 4e, Potassium
(7-chloro-4-azaindol-3-yl)-oxoacetate was prepared by the same
method as Precursor 4a, starting from Methyl
(7-chloro-4-azaindol-3-yl)-oxoacetate, Precursor 3f. MS m/z:
(M+H).sup.+ of the corresponding acid of compound 4e (M-K.sup.+
H)+calcd for C.sub.9H.sub.6ClN.sub.2O.sub.3: 225.01; found 225.27.
HPLC retention time: 0.33 minutes (column A).
EXAMPLE 37
[0551] 151
[0552] The standard coupling procedures described earlier were used
to couple intermediates 4ae and intermediate 1 with procedures to
provide Example 37 ret. time=0.65 min (column G, solvent A) Exact
Mass: 419.11
EXAMPLE 38
[0553] 152
[0554] Standard Stille coupling conditions as described earlier
were used to provide Example 38 after coupling with 2-tributyl
stannyl thiazole. ret. time=0.78 min (column G, solvent a) Exact
Mass: 468.14
[0555] LCMS Conditions:
[0556] Solvent A: 10% MeOH --90% H.sub.2O-0.1% TFA
[0557] Solvent B: 90% MeOH --10% H.sub.2O-0.1% TFA
[0558] Column: XTERRA C18 S7 3.0.times.50 mm
[0559] Start % B=0
[0560] Final % B=100
[0561] Gradient Time=2 min
[0562] Flow Rate=5 m/min
[0563] Wavelength=220
EXAMPLE 39
[0564] 153
[0565] Using the procedures described herein the title compound was
prepared:
[0566] .sup.1H NMR: (CD.sub.3OD) .delta. 9.23 (s, 1H), 8.59 (d,
J=7.0, 1H), 8.37 (s, 1H), 8.32 (d, J=8.5, 1H), 8.01-7.96 (dd
overlapped with d, 2H), 7.90 (s, 1H), 7.76 (app t, 1H), 7.28 (d,
J=7.0, 1H), 4.09 (s overlapped with m, 7H), 3.98 (m, 2H), 3.88 (m,
2H), 2.56 (s, 3H); LC/MS: (ES+) m/z (M+H).sup.+=497; HPLC
R.sub.t=0.937.
EXAMPLE 40
[0567] 154 155
[0568] 25 mgs (0.085147 mmol) of
(4-Methoxy-7-[1,2,4]oxadiazol-3-yl-1H-pyr-
rolo[2,3-c]pyridin-3-yl)-oxo-acetyl chloride and 19.4 mgs (0.085
mmol) of 2-Methyl-4-piperazin-1-yl-quinoline were suspended in 2 mL
of dichloromethane in a vial and cooled to -10.degree. C.
Diisopropylethylamine (22.2 .mu.L, 1.5equivalents) was then added
and the reaction was shaken for 10 min. The reaction was allowed to
stand. A pale yellow precipitate formed after 10 min. After
standing for two hours total, the suspension was dissolved with 20
mL of dichloromethane and 15 mL water. Extraction and then
reextraction with 10 mL of dichloromethane provided combined
organic extracts which were dried over anhydrous Magnesium Sulfate,
filtered and concentrated in vacuo to provide 20 mgs of the desired
product [M+H]+=485 and LC purity=87% at 215 nM and a ten minute
elution.
EXAMPLES 41-42
[0569] 156
[0570] A well of a standard 96 well plate was loaded with 1 mL of
dichloromethane then 1.1 eq of the corresponding piperazine and
then acid chloride (1.1 eq, 0.0470 to 0.0532 mmol) were then added.
Next 5eqs of Hunig's base (diisopropylethylamine) were added and
the plate shaken overnight at ambient temperature. Two equivalents
of PAMPS (n-propylaminomethylolystyrene, 1/mmol per gram) were
added for each equivalent of acid chloride and the reaction mixture
shaken overnight. The wells were agitated by adding, pipetting, and
re-adding 0.5 mL citric acid about ten times. The contents of the
well was passed through anhydrous MgSO4, and the products either
used as formed or purified by passage over SiO2 using 9:1
ethylacetate: methanol.
[0571] Data for Examples 40-42
[0572] 10 minute HPLC method for example 40
[0573] 1. Apparatus and Reagents
[0574] 1.1 Common Apparatus
[0575] 0.1% Trifluoroacetic acid (aq)--Mobile phase "A"
[0576] 0.1% Trifluoroacetic acid (acetonitrile)--Mobile phase
"B"
[0577] Phenomenex Luna C8 (2) 100.times.2.0 mm, 3 .mu.m column
[0578] Waters Millennium.sup.32.TM. Chromatography Data System
(V3.2 or better)
[0579] 1.2 Instrumentation
[0580] Waters 2790 LC system ("LC19"), comprising:
[0581] Waters 2790 Separations Module
[0582] Waters 2487 Dual Wavelength Absorbance Detector--wavelength
set at 215 nm.
[0583] 2. Instrument Parameters
[0584] LC Conditions
[0585] Minutes
[0586] The dashed line represents re-equilibration. Overall run
time is 13.5 minutes, the mass spectrometer and Millennium.sup.32
captures the first 10 minutes of the run.
[0587] Flow rate=0.3 ml/min
[0588] Run time=13.5 minutes
4 Gradient: Time (mins) % Organic 0.00 5 6.30 95 9.50 95 9.70 5
13.5 5
[0589] 3. Integration and Reporting
[0590] Data is integrated using Millennium and reported via the
Millennium software.
[0591] 2.5 Minute HPLC Method for Examples 41 and 42
[0592] 4. Apparatus and Reagents
[0593] 4.1 Common Apparatus
[0594] 0.1% Trifluoroacetic acid (aq)--Mobile phase "A"
[0595] 0.1% Trifluoroacetic acid (acetonitrile)--Mobile phase
"B"
[0596] Hypersil BDS C18 column 5 um, 2.1.times.50 mm
[0597] Micromass MassLynx.TM. Operating Software with OpenLynx.TM.
Browser Option (V3.5 or better)
[0598] Waters Millennium.sup.32.TM. Chromatography Data System
(V3.2 or better)
[0599] 4.2 Instrumentation
[0600] 4.2.1 Micromass Single Quadrupole LCMS systems ("MS1",
"MS4", "MS6" or "MS7"), comprising:
[0601] Agilent HP1100 LC system comprising the following
modules:
[0602] G1315A Diode Array Detector or G1314A Single Wavelength UV
Detector
[0603] G1312A Binary Pump with Pulse Dampener and Mixer fitted
[0604] G1316A Vacuum Degasser (optional)
[0605] G1316A Column Oven (optional)
[0606] Polymer LabsPL1000 Evaporative Light Scattering Detector
(ELSD) with either
[0607] CTC Analytics HTC PAL Autosampler or
[0608] Gilson 215 Single Probe Autosampler with either
[0609] Micromass Platform LC or
[0610] Micromass ZMD single quadrupole mass spectrometer
[0611] 4.2.2 Micromass LCT Systems ("M55", "MS8" or "MS9"),
comprising:
[0612] MS5
[0613] Agilent HP1100 LC system comprising the following
modules:
[0614] G1314A Single Wavelength UV Detector G1312A Binary Pump with
Pulse Dampener and Mixer fitted
[0615] CTC Analytics HTC PAL Autosampler
[0616] Micromass LCT with Z-spray Interface
[0617] MS8
[0618] Waters 600 Binary Pump
[0619] 8.times. Waters 2487 Dual Wavelength Detector
[0620] Gilson 215 Multiprobe 8-way Autosampler
[0621] Micromass LCT with MUX.TM. 8-way interface
[0622] MS9
[0623] Waters 1525 Binary Pump
[0624] 1.times.2488 Dual Wavelength 8-way detector
[0625] CTC Analytics HTS PAL Autosampler with 4-fold injection
valve
[0626] Micromass LCT with MUX.TM. 5-way interface
[0627] 5. LC Conditions
[0628] 5.1.1 LC Conditions--for MS8.
[0629] Flow rate=8.0 ml/min--split 8 ways to deliver 1 ml/min
through all 8 lines
5 Time (mins) % B 0 0 1.80 95 2.10 95 2.30 0 2.90 0
[0630] 5.1.2 LC Conditions--for MS9.
[0631] Flow rate=4.0 mllmin--split 4 ways to deliver 1 ml/min
through all 4 lines
6 Time (mins) % B 0 0 1.80 95 2.10 95 2.30 0 2.39 0
[0632] 5.2 Mass Spectrometer Conditions
[0633] Data is typically collected over the range m/z 150 to 850 at
a sampling rate of 2 scans per second (1 scan per 1.2 seconds per
line on MS8).
[0634] 6. Integration and Reporting
[0635] Data is integrated using OpenLynx and reported via the
OpenLynx Browser software.
7 HPLC HLPC Ret. Example # MW Method Time Mass spec (purity)
Example 40 484.49 10 Min. 4.23 min. 485.25 (100%) Example 41 434.43
2.5 Min. 1.03 min. 435.25 (100%) Example 42 453.86 2.5 Min. 1.56
min. 454.22 (100%) Intermediate 9 (HCl salt) 157
[0636] Hydrochloride salt of intermediate 9 was prepared from the
corresponding 1-hydroxyphthalazine by conversion to the 1-chloro
derivative (neat POCl.sub.3, 130.degree. C.), followed by
condensation with tert-butyl 1-piperazinecarboxylate (Et.sub.3N,
nBuOH, 130.degree. C.) and then deprotection (4N HCl in
1,4-dioxane, r.t.); HCl salt .sup.1H NMR: (CD.sub.3OD) .delta. 9.83
(s, 1H), 8.54-8.49 (m, 2H), 8.38 (app t, 1H), 8.28 (app t, 1H),
4.01 (b s, 4H), 3.58 (b s, 4H); Analytical HPLC method: Start %
B=0, Final % B=100, Gradient time=2 min, Flow Rate=5 mL/min,
Column: Xterra C18 S7 30.0.times.50 mm, LC/MS: (ES+) f/z
(M+H).sup.+=215.12, HPLC R.sub.t=0.083. 158
[0637] Hydrochloride salt of Intermediate 10 was prepared in a
similar manner as Intermediate 9, except that the following
chlorination conditions were used: POCl.sub.3, N,N-diethylaniline,
benzene, reflux (Connolly, D. J.; Guiry, P. J. Synlett 2001,
1707.); HCl salt .sup.1H NMR: (CD.sub.3OD) .delta. 8.26 (d, J=10,
1H), 8.05 (app t, 1H), 7.83 (d, J=5, 1H), 7.76 (app t, 1H), 4.53 (b
s, 4H), 3.56 (b s, 4H), 2.75 (s, 3H); Analytical HPLC method: Start
% B=0, Final % B=100, Gradient time=2 min, Flow Rate=5 mL/min,
Column: Xterra C18 S7 3.0.times.50 mm, LC/MS: (ES+) m/z
(M+H).sup.+=229.40, HPLC R.sub.t=0.077. 159
[0638] Hydrochloride salt of Intermediate 11 was isolated as a side
product during the preparation of the corresponding 1-chloro
analog; Analytical HPLC method: Start % B=0, Final % B=100,
Gradient time=2 min, Flow Rate=5 mL/min, Column: Xterra C18 S7
3.0.times.50 mm, LC/MS: (ES+) m/z (M+H).sup.+=245.13, HPLC
R.sub.t=0.523.
EXAMPLE 44
[0639] 160
[0640] Example 44 was prepared in a similar manner as Example 43
using EDC/DMAP as the coupling ragents, and purification by
preparative reverse phase HPLC. Preparative reverse phase HPLC
separation method: Start % B=0, Final % B=100, Gradient time=6 min,
Flow Rate=30 mL/min, Column: Xerra Prep MS C18 5 uM 19.times.50 mm,
Fraction Collection: 2.88-3.49 min; .sup.1H NMR: (CD.sub.3OD)
.delta. 8.74 (s, 1H), 8.29 (d, J=10, 1H), 8.21 (s, 1H), 8.05 (d,
J=10, 1H), 7.83-7.76 (m, 2H), 7.67 (d, J=10, 1H), 6.91 (d, J=10,
1H), 4.49 (b s, 2H), 4.37 (b s, 2H), 4.00 (b s, 5H), 3.80 (b s,
2H); Analytical HPLC method: Start % B=0, Final % B=100, Gradient
time=2 min, Flow Rate=5 mL/min, Column: Xterra C18 S7 30.0.times.50
mm; LC/MS: (ES+) m/z (M+H).sup.+=441.27, HPLC R.sub.t=1.123.
EXAMPLE 45
[0641] 161
[0642] Example 45 was prepared in a similar manner as Example 9
using HATU/DMAP as the coupling reagents. Preparative reverse phase
HPLC separation method: Start % B=0, Final % B=100, Gradient time=6
min, Flow Rate=30 mL/min, Column: Xerra Prep MS C18 5 uM
19.times.50 mm, Fraction Collection: 2.72-3.30 min; .sup.1H NMR:
(CD.sub.3OD) .delta. 8.74 (s, 1H), 8.44 (s, 1H), 8.30 (d, J=10,
1H), 8.19 (s, 1H), 8.06 (t, J=10, 1H), 7.83-7.77 (m, 2H), 4.49 (b
s, 2H), 4.38 (b s, 2H), 4.12 (b s, 3H), 4.01 (b s, 2H), 3.82 (b s,
2H); Analytical HPLC method: Start % B=0, Final % B=100, Gradient
time=2 min, Flow Rate=5 mL/min, Column: Xterra C18 S7 3.0.times.50
mm; LC/MS: (ES+) m/z (M+H).sup.+=442.24, HPLC R.sub.t=1.053.
EXAMPLE 46
[0643] 162
[0644] Example 46 was prepared in a similar manner as Example 36
using HATU/DMAP as the coupling reagents. Preparative reverse phase
HPLC separation method: Start % B=0, Final % B=100, Gradient time=6
min, Flow Rate=30 mL/min, Column: Xerra Prep MS C18 5 uM
19.times.50 mm, Fraction Collection: 3.14-3.74 min; .sup.1H NMR:
(CD.sub.3OD) .delta. 9.23 (s, 1H), 8.74 (s, 1H), 8.35 (s, 1H), 8.31
(d, J=5, 1H), 8.06 (app t, 1H), 7.86 (s, 1H), 7.83 (d, J=5, 1H),
7.78 (app t, 1H), 4.52 (b s, 2H), 4.41 (b s, 2H), 4.06 (s, 3H),
4.04 (b s, 2H), 3.84 (b s, 2H), 2.55 (s, 3H); Analytical HPLC
method: Start % B=0, Final % B=100, Gradient time=2 min, Flow
Rate=5 mL/min, Column: Xterra C18 S7 3.0.times.50 mm; LC/MS: (ES+)
m/z (M+H).sup.+=498.19, HPLC R.sub.t=0.910.
EXAMPLE 47
[0645] 163
[0646] Example 47 was prepared in a similar manner as Example 46.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=30 mL/min, Column:
Xerra Prep MS C18 5 uM 19.times.50 mm, Fraction Collection:
3.08-3.40 min; .sup.1H NMR: (CD.sub.3OD) .delta. 9.37 (s, 1H), 8.74
(s, 1H), 8.36 (s, 1H), 8.33 (s, 1H), 8.30 (d, J=10, 1H), 8.05 (app
t, 1H), 7.90 (s, 1H), 7.83 (d, J=10, 1H), 7.77 (app t, 1H), 4.50 (b
s, 2H), 4.39 (b s, 2H), 4.07 (s, 3H), 4.04 (b s, 2H), 3.84 (b s,
2H); Analytical HPLC method: Start % B=0, Final % B=100, Gradient
time=2 min, Flow Rate=5 mL/min, Column: Xterra C18 S7 30.0.times.50
mm; LC/MS: (ES+) m/z (M+H).sup.+=484.18, HPLC R.sub.t=0.843.
EXAMPLE 48
[0647] 164
[0648] Example 48 was prepared in a similar manner as Example 46.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=30 mL/min, Column:
Xerra Prep MS C18 5 uM 19.times.50 mm, Fraction Collection:
3.18-3.79 min; .sup.1H NMR: (CD.sub.3OD) .delta. 8.91 (s, 1H), 8.74
(s, 1H), 8.47 (s, 1H), 8.29 (d, J=10, 1H), 8.20 (d, J=5, 1H), 8.05
(app t, 1H), 8.00 (s, 1H), 7.83 (d, J=5, 1H), 7.77 (app t, 1H),
4.50 (b s, 2H), 4.40 (b s, 2H), 4.04 (b s, 2H), 3.89 (b s, 2H);
Analytical HPLC method: Start % B=0, Final % B=100, Gradient time=2
min, Flow Rate=5 mL/min, Column: Xterra C18 S7 3.0.times.50 mm;
LC/MS: (ES+) m/z (M+H).sup.+=472.14, HPLC R.sub.t=1.007.
EXAMPLE 49
[0649] 165
[0650] Example 49 was prepared from Example 45. Preparative reverse
phase HPLC separation method: Start % B=0, Final % B=100, Gradient
time=5 min, Flow Rate=25 mL/min, Column: Xterra Prep 19.times.50 mm
S5, Fraction Collection: 2.29-2.98 min; .sup.1H NMR: (CD.sub.3OD)
.delta. 8.75 (s, 1H), 8.54 (s, 1H), 8.32 (d, J=5, 1H), 8.14 (s,
1H), 8.06 (d, J=5, 1H), 7.84 (d, J=10, 1H), 7.78 (b s, 1H), 4.52 (b
s, 2H), 4.41 (b s, 2H), 4.15 (b s, 3H), 4.04 (b s, 2H), 3.86 (b s,
2H); Analytical HPLC method: Start % B=0, Final % B=100, Gradient
time=2 min, Flow Rate=5 mL/min, Column: Xterra C18 S7 30.0.times.50
mm; LC/MS: (ES+) m/z (M+H).sup.+=460.28, HPLC R.sub.t=0.763.
EXAMPLE 50
[0651] 166
[0652] Example 50 was prepared from Example 49. .sup.1H NMR:
(CD.sub.3OD) .delta. 8.64 (s, 1H), 8.32 (s, 1H), 8.10 (s, 2H), 7.85
(d, J=5, 2H), 7.60-7.57 (m, 1H), 4.11 (s, 3H), 4.01 (b s, 2H), 3.99
(b s, 2H), 3.90 (b s, 2H), 3.73 (b s, 2H); Analytical HPLC method:
Start % B=0, Final % B=100, Gradient time=2 min, Flow Rate=5
mL/min, Column: Xterra C18 S7 3.0.times.50 mm; LC/MS: (ES+) m/z
(M+H).sup.+=461.17, HPLC R.sub.t=0.743.
EXAMPLE 51
[0653] 167
[0654] Example 51 was prepared in a similar manner as Example 46.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=30 mL/min, Column:
Xerra Prep MS C18 5 uM 19.times.50 mm, Fraction Collection:
1.62-2.06 min; .sup.1H NMR: (CD.sub.3OD) .delta. 8.74 (s, 1H), 8.39
(s, 1H), 8.31 (d, J=10, 1H), 8.14 (s, 2H), 8.06 (app t, 1H), 7.92
(b s, 1H), 7.83 (d, J=5, 1H), 7.78 (app t, 1H), 4.52 (b s, 2H),
4.40 (b s, 2H), 4.09 (s, 3H), 4.04 (b s, 2H), 3.85 (b s, 2H);
Analytical HPLC method: Start % B=0, Final % B=100, Gradient time=2
min, Flow Rate=5 mL/min, Column: Xterra C18 S7 3.0.times.50 mm;
LC/MS: (ES+) m/z (M+H).sup.+=484.18, HPLC R.sub.t=0.893.
EXAMPLE 52
[0655] 168
[0656] Example 52 was prepared in a similar manner as Example 46.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=30 mL/min, Column:
Xerra Prep MS C18 5 uM 19.times.50 mm, Fraction Collection:
3.13-3.69 min; .sup.1H NMR: (CD.sub.3OD) .delta. 8.86 (s, 1H), 8.74
(s, 1H), 8.39 (s, 1H), 8.31 (d, J=10, 1H), 8.06 (app t, 1H), 7.98
(s, 1H), 7.97 (s, 1H), 7.83 (d, J=5, 1H), 7.78 (app t, 1H), 4.51 (b
s, 2H), 4.41 (b s, 2H), 4.09 (s, 3H), 4.04 (b s, 2H), 3.85 (b s,
2H); Analytical HPLC method: Start % B=0, Final % B=100, Gradient
time=2 min, Flow Rate=5 mL/min, Column: Xteffa C18 S7 3.0.times.50
mm; LC/MS: (ES+) m/z (M+H).sup.+=484.10, HPLC R.sub.t=0.990.
EXAMPLE 53
[0657] 169
[0658] Example 53 was prepared in a similar manner as Example 9
using HATU/DMAP as the coupling reagents. .sup.1H NMR: (CD.sub.3OD)
.delta. 8.72 (s, 1H), 8.35 (s, 1H), 8.26 (d, J=10, 1H), 8.02 (app
t, 1H), 7.84 (d, J=5, 1H), 7.80 (s, 1H), 7.74 (app t, 1H), 4.41 (b
s, 2H), 4.29 (b s, 2H), 4.01 (b s, 5H), 3.80 (b s, 2H); Analytical
HPLC method: Start % B=0, Final % B=100, Gradient time=2 min, Flow
Rate=5 mL/min, Column: Xterra C18 S7 3.0.times.50 mm; LC/MS: (ES+)
m/z (M+H).sup.+=496.97, HPLC R.sub.t=0.773.
EXAMPLE 54
[0659] 170
[0660] Example 54 was prepared in a similar manner as Example 46.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=30 mL/min, Column:
Xerra Prep MS C18 5 uM 19.times.50 mm, Fraction Collection:
3.70-4.16 min; .sup.1H NMR: (CD.sub.3OD) .delta. 9.51 (s, 1H), 8.86
(b s, 2H), 8.48 (d, J=10, 1H), 8.39 (s, 1H), 8.25 (t, J=10, 1H),
8.21 (d, J=10, 1H), 7.98 (b s, 2H), 4.13 (s, 3H), 4.09 (b s, 2H),
4.07 (s, 2H), 3.95 (b s, 2H), 3.86 (b s, 2H); Analytical HPLC
method: Start % B=0, Final % B=100, Gradient time=2 min, Flow
Rate=5 mL/min, Column: Waters Atlantis 4.6.times.50 mm C18 5 um;
LC/MS: (ES+) m/z (M+H).sup.+=484.11, HPLC R.sub.t=1.213.
EXAMPLE 55
[0661] 171
[0662] Example 55 was prepared in a similar manner as Example 46.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=25 mL/min, Column:
Xterra 5 uM 19.times.50 mm, Fraction Collection: 3.17-3.87 min;
.sup.1H NMR: (CD.sub.3OD) .delta. 9.56 (s, 1H), 9.24 (s, 1H), 8.51
(d, J=10, 1H), 8.43 (d, J=5, 1H), 8.34 (s, 1H), 8.29 (app t, 1H),
8.24 (app t, 1H), 7.87 (s, 1H), 4.09 (b s, 5H), 3.99 (b s, 2H),
3.89 (b s, 2H), 3.87 (b s, 2H), 2.55 (s, 3H); Analytical HPLC
method: Start % B=0, Final % B=100, Gradient time=2 min, Flow
Rate=5 mL/min, Column: Xterra C18 S7 30.0.times.50 mm; LC/MS: (ES+)
m/z (M+H).sup.+=498.12, HPLC R.sub.t=0.907.
EXAMPLE 56
[0663] 172
[0664] Example 56 was prepared in a similar manner as Example 46.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=30 mL/min, Column:
Xterra 5 uM 19.times.50 mm, Fraction Collection: 3.86-4.47 min;
.sup.1H NMR: (CD.sub.3OD) .delta. 9.52 (s, 1H), 8.92 (s, 1H), 8.47
(s, 1H), 8.39 (d, J=5, 1H), 8.26 (t, J=10, 1H), 8.22 (b s, 2H),
8.00 (s, 2H), 4.10 (b s, 2H), 3.96 (b s, 2H), 3.89 (b s, 2H), 3.87
(b s, 2H); Analytical HPLC method: Start % B=0, Final % B=100,
Gradient time=2 min, Flow Rate=5 mL/min, Column: Waters Atlantis
4.6.times.50 mm C18 5 um; LC/MS: (ES+) m/z (M+H).sup.+=472.08, HPLC
R.sub.t=1.313.
EXAMPLE 57
[0665] 173
[0666] Example 57 was prepared in a similar manner as Example 46.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=30 mL/min, Column:
Xterra 5 uM 19.times.50 mm, Fraction Collection:
[0667] 3.88-4.47 min; .sup.1H NMR: (CD.sub.3OD) .delta. 9.23 (s,
1H), 8.47 (d, J=10, 1H), 8.35 (s, 1H), 8.31 (s, 1H), 8.19 (t, J=5,
1H), 7.88 (s, 1H), 7.82 (s, 1H), 4.22 (s, 3H), 4.13 (b s, 2H), 4.10
(s, 3H), 4.00 (b s, 2H), 3.89 (b s, 2H), 3.85 (b s, 2H), 2.55 (s,
3H); Analytical HPLC method: Start % B=0, Final % B=100, Gradient
time=2 min, Flow Rate=5 mL/min, Column: Waters Atlantis
4.6.times.50 mm C18 5 um; LC/MS: (ES+) m/z (M+H).sup.+=528.29, HPLC
R.sub.t=1.543.
EXAMPLE 58
[0668] 174
[0669] Example 58 was isolated as a side product of Example 57.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=30 mL/min, Column:
Xterra 5 uM 19.times.50 mm, Fraction Collection: 4.51-4.89 min;
.sup.1H NMR: (CD.sub.3OD) .delta. 9.25 (s, 1H), 8.35 (s, 1H), 8.33
(s, 1H), 8.10 (d, J=10, 1H), 7.94 (t, J=5, 1H), 7.89 (s, 1H), 7.86
(s, 1H), 4.11 (s, 3H), 4.01 (b s, 2H), 3.94 (b s, 2H), 3.82 (b s,
2H), 3.75 (b s, 2H), 2.56 (s, 3H); Analytical HPLC method: Start %
B=0, Final % B=100, Gradient time=2 min, Flow Rate=5 mL/min,
Column: Waters Atlantis 4.6.times.50 mm C18 5 um; LC/MS: (ES+) m/z
(M+H).sup.+=514.27, HPLC R.sub.t=1.643.
EXAMPLE 59
[0670] 175
[0671] Example 59 was prepared in a similar manner as Example 46.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=30 mL/min, Column:
Xterra 5 uM 19.times.50 mm, Fraction Collection: 3.20-4.27 min;
.sup.1H NMR: (CD.sub.3OD) .delta. 8.91 (s, 1H), 8.47 (s, 1H), 8.25
(d, J=10, 1H), 8.19 (s, 1H), 8.02-7.99 (m, 2H), 7.75-7.70 (m, 2H),
4.48 (b s, 2H), 4.37 (b s, 2H), 4.03 (b s, 2H), 3.89 (b s, 2H),
2.70 (s, 3H); Analytical HPLC method: Start % B=0, Final % B=100,
Gradient time=2 min, Flow Rate=5 mL/min, Column: Xterra
3.0.times.50 mm; LC/MS: (ES+) m/z (M+H).sup.+=486.09, HPLC
R.sub.t=1.160.
EXAMPLE 60
[0672] 176
[0673] Example 60 was prepared in a similar manner as Example 46.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=30 mL/min, Column:
Xterra 5 uM 19.times.50 mm, Fraction Collection: 3.04-3.65 min;
.sup.1H NMR: (CD.sub.3OD) .delta. 9.21 (s, 1H), 8.33 (s, 1H), 8.26
(d, J=5, 1H), 8.00 (t, J=5, 1H), 7.84 (s, 1H), 7.75-7.69 (m, 2H),
4.49 (b s, 2H), 4.37 (b s, 2H), 4.05 (s, 3H), 4.02 (b s, 2H), 3.83
(b s, 2H), 2.69 (s, 3H), 2.55 (s, 3H); Analytical HPLC method:
Start % B=0, Final % B=100, Gradient time=2 min, Flow Rate=5
mL/min, Column: Xterra 3.0.times.50 mm; LC/MS: (ES+) m/z
(M+H).sup.+=512.13, HPLC R.sub.t=1.090.
EXAMPLE 61
[0674] 177
[0675] Example 61 was prepared in a similar manner as Example 46.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=30 mL/min, Column:
Xterra 5 uM 19.times.50 mm, Fraction Collection: 2.90-3.97 min;
.sup.1H NMR: (CD.sub.3OD) .delta. 8.85 (s, 1H), 8.38 (s, 1H), 8.27
(d, J=5, 1H), 8.01 (t, J=5, 1H), 7.98 (s, 1H), 7.95 (s, 1H),
7.75-7.70 (m, 2H), 4.49 (b s, 2H), 4.38 (b s, 2H), 4.09 (s, 3H),
4.03 (b s, 2H), 3.84 (b s, 2H), 2.70 (s, 3H); Analytical HPLC
method: Start % B=0, Final % B=100, Gradient time=2 min, Flow
Rate=5 mL/min, Column: Xterra 3.0.times.50 mm; LC/MS: (ES+) m/z
(M+H).sup.+=498.15, HPLC R.sub.t=0.983.
EXAMPLE 62
[0676] 178
[0677] Example 62 was prepared in a similar manner as Example 53
using EDC/HOBT as the coupling reagents. Preparative reverse phase
HPLC separation method: Start % B=0, Final % B=100, Gradient time=6
min, Flow Rate=30 mL/min, Column: Xterra 5 uM 19.times.50 mm,
Fraction Collection: 2.25-2.65 min; .sup.1H NMR: (CD.sub.3OD)
.delta. 9.55 (s, 1H), 8.70 (d, J=5, 1H), 8.50 (d, J=5, 1H), 8.40
(d, J=10, 1H), 8.27 (app t, 1H), 8.22 (app t, 1H), 8.08 (s, 1H),
4.11 (s, 3H), 4.09 (b s, 2H), 3.96 (b s, 4H), 3.91 (b s, 2H);
Analytical HPLC method: Start % B=0, Final % B=100, Gradient time=2
min, Flow Rate=5 mL/min, Column: Waters Atlantis 4.6.times.50 mm
C18 5 um; LC/MS: (ES+) m/z (M+H).sup.+=497.11, HPLC
R.sub.t=0.910.
EXAMPLE 63
[0678] 179
[0679] Example 63 was prepared from Example 62 and used as crude
for the preparation of Example 64; Analytical HPLC method: Start %
B=0, Final % B=100, Gradient time=2 min, Flow Rate=5 mL/min,
Column: Waters Atlantis 4.6.times.50 mm C18 5 um; LC/MS: (ES+) m/z
(M+H).sup.+=442.24, HPLC R.sub.t=1.133.
EXAMPLE 64
[0680] 180
[0681] Example 64 was prepared from Example 63. Preparative reverse
phase HPLC separation method: Start % B=0, Final % B=100, Gradient
time=6 min, Flow Rate=30 mL/min, Column: Xterra 5 uM 19.times.50
mm, Fraction Collection: 2.76-3.14 min; .sup.1H NMR: (CD.sub.3OD)
.delta. 9.46 (s, 1H), 8.44 (d, J=5, 1H), 8.33 (s, 1H), 8.21 (t,
J=5, 1H), 8.13 (s, 1H), 7.89 (d, J=5, 1H), 7.72-7.70 (m, 1H), 4.21
(b s, 2H), 4.14 (s, 3H), 4.08 (b s, 2H), 3.90 (b s, 2H), 3.83 (b s,
2H); Analytical HPLC method: Start % B=0, Final % B=100, Gradient
time=2 min, Flow Rate=5 mL/min, Column: Waters Atlantis
4.6.times.50 mm C18 5 um; LC/MS: (ES+) m/z (M+H).sup.+=460.21, HPLC
R.sub.t=1.077. 181
[0682] Hydrochloride salt of Intermediate 12 was prepared from
3-aminol-bromoisoquinoline in a similar manner as Intermediate 9.
Analytical HPLC method: Start % B=0, Final % B=100, Gradient time=2
min, Flow Rate=5 mL/min, Column: Xterra MS C18 S7 3.0.times.50 mm;
LC/MS: (ES+) m/z (M+H).sup.+=229.12, HPLC R.sub.t=0.343. 182
[0683] Hydrochloride salt of Intermediate 13 was prepared from
1,3-dichloroisoquinoline in a similar manner as Intermediate 2.
.sup.1H NMR: (CD.sub.3OD, 300 MHz) .delta. 8.15 (d, J=8.4, 1H),
7.84 (d, J=8.1, 1H), 7.74 (app t, 1H), 7.63 (app t, 1H), 7.51 (s,
1H), 3.71-3.68 (m, 4H), 3.54-3.50 (m, 4H); Analytical HPLC method:
Start % B=0, Final % B=100, Gradient time=2 min, Flow Rate=5
mL/min, Column: Xterra C18 4.6.times.50 mm C18 5 um; LC/MS: (ES+)
m/z (M+H).sup.+=248.02, 250.02, HPLC R.sub.t=1.253.
EXAMPLE 65
[0684] 183
[0685] Example 65 was prepared in a similar manner as Example 46.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=45 mL/min, Column:
phenomenex-Luna 30.times.50 mm S5, Fraction Collection: 4.51-4.92
min; .sup.1H NMR: (CD.sub.3OD) .delta. 9.22 (s, 1H), 8.77 (s, 1H),
8.31 (s, 1H), 8.13 (d, J=8, 1H), 7.88 (d, J=8, 1H), 7.84 (s, 1H),
7.70 (t, J=8, 1H), 7.56 (t, J=7.5, 1H), 3.98 (s, 3H), 3.73 (m, 4H),
3.54 (m, 4H), 2.58 (s, 3H); Analytical HPLC method: Start % B=0,
Final % B=100, Gradient time=2 min, Flow Rate=5 mL/min, Column:
Xterra MS C18 S7 3.0.times.50 mm; LC/MS: (ES+) m/z
(M+H).sup.+=512.20, HPLC R.sub.t=1.277.
EXAMPLE 66
[0686] 184
[0687] Example 66 was prepared in a similar manner as Example 65.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=30 mL/min, Column:
Xterra 19.times.50 mm S5, Fraction Collection: 5.76-6.32 min;
.sup.1H NMR: (CD.sub.3OD) .delta. 9.24 (s, 1H), 8.34 (s, 1H), 8.16
(d, J=8.5, 1H), 7.89 (s, 1H), 7.79 (d, J=8, 1H), 7.69 (app t, 1H),
7.58 (app t, 1H), 7.41 (s, 1H), 4.10 (s, 3H), 4.03 (m, 2H), 3.77
(m, 2H), 3.61 (m, 2H), 3.50 (m, 2H), 2.56 (s, 3H); Analytical HPLC
method: Start % B=0, Final % B=100, Gradient time=2 min, Flow
Rate=5 ml/min, Column: Xterra 4.6.times.50 mm C18 5 um; LC/MS:
(ES+) m/z (M+H).sup.+=530.99, 532.98, HPLC R.sub.t=1.840.
EXAMPLE 67
[0688] 185
[0689] Example 67 was prepared in a similar manner as Example 65.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=30 mL/min, Column:
Xterra 19.times.50 mm S5, Fraction Collection: 5.59-6.20 min;
.sup.1H NMR: (CD.sub.3OD) .delta. 8.86 (s, 1H), 8.38 (s, 1H), 8.18
(d, 1H), 7.98 (overlapping s, 2H), 7.79 (d, J=8.5, 1H), 7.69 (app
t, 1H), 7.59 (app t, 1H), 7.41 (s, 1H), 4.13 (s, 3H), 4.05 (m, 2H),
3.80 (m, 2H), 3.62 (m, 2H), 3.50 (m, 2H); Analytical HPLC method:
Start % B=0, Final % B=100, Gradient time=2 min, Flow Rate=5
mL/min, Column: Xterra 4.6.times.50 mm C18 5 um; LC/MS: (ES+) m/z
(M+H).sup.+=517.00, 518.90, HPLC R.sub.t=1.903.
EXAMPLE 68
[0690] 186
[0691] Example 68 was prepared in a similar manner as Example 65.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=30 mL/min, Column:
Xterra 19.times.50 mm S5, Fraction Collection: 5.79-6.39 min;
.sup.1H NMR: (CD.sub.3OD) .delta. 8.92 (s, 1H), 8.44 (s, 1H), 8.12
(m, 1H), 8.06 (s, 1H), 7.99 (d, H), 7.78 (app t, 1H), 7.69 (m, 1H),
7.58 (m, 1H), 7.40 (d, 1H), 4.10-3.55 (m, 8H); Analytical HPLC
method: Start % B=0, Final % B=100, Gradient time=2 min, Flow
Rate=5 mL/min, Column: Xterra 4.6.times.50 mm C18 5 um; LC/MS:
(ES+) m/z (M+H).sup.+=504.95, 506.95, HPLC R.sub.t=1.907.
EXAMPLE 69
[0692] 187
[0693] Example 69 was prepared from condensation of the Example 53
with 3-cyano-1,2,4-triazole at 150.degree. C. Preparative reverse
phase HPLC separation method: Start % B=0, Final % B=100, Gradient
time=6 min, Flow Rate=30 mL/min, Column: Xterra 19.times.50 mm S5,
Fraction Collection: 3.46-3.96 min; .sup.1H NMR: (CD.sub.3OD)
.delta. 9.52 (s, 1H), 8.75 (s, 1H), 8.37 (s, 1H), 8.31 (d, J=8.5,
1H), 8.06 (app t, 1H), 7.94 (s, 1H), 7.83 (d, 1H), 7.78 (app t,
1H), 4.51 (m, 2H), 4.40 (m, 2H), 4.09 (s, 3H), 4.03 (m, 2H), 3.85
(m, 2H); Analytical HPLC method: Start % B=0, Final % B=100,
Gradient time=2 min, Flow Rate=5 mL/min, Column: Xterra
4.6.times.50 mm C18 5 um; LC/MS: (ES+) m/z (M+H).sup.+=509.01, HPLC
R.sub.t=1.140.
EXAMPLE 70
[0694] 188
[0695] Example 70 was prepared from Example 69 using concentrated
methanolic hydrogen chloride. Preparative reverse phase HPLC
separation method: Start % B=0, Final % B=100, Gradient time=6 min,
Flow Rate=30 mL/min, Column: Xterra 19.times.50 mm S5, Fraction
Collection: 3.64-3.84 min; .sup.1H NMR: (CD.sub.3OD) .delta. 9.40
(s, 1H), 8.77 (s, 1H), 8.42 (s, 1H), 8.33 (d, J=8.6, 1H), 8.07 (d,
J=8.2, 1H), 7.91 (s, 1H), 7.85 (d, J=8.6, 1H), 7.80 (app t, 1H),
4.55-4.53 (m, 2H), 4.44-4.42 (m, 2H), 4.10 (s, 3H), 4.07-4.05 (m,
2H), 3.89-3.87 (m, 2H); Analytical HPLC method: Start % B=0, Final
% B=100, Gradient time=2 min, Flow Rate=5 mL/min, Column: Xterra
4.6.times.50 mm C18 5 um; LC/MS: (ES+) m/z (M+H).sup.+=527.05, HPLC
R.sub.t=1.050.
EXAMPLE 71
[0696] 189
[0697] Example 71 was prepared in a similar manner as Example 53.
Analytical HPLC method: Start % B=0, Final % B=100, Gradient time=2
min, Flow Rate=5 mL/min, Column: Xterra MS C18 S7 30.0.times.50 mm;
LC/MS: (ES+) m/z (M+H).sup.+=482, 484, HPLC R.sub.t=0.980.
EXAMPLE 72
[0698] 190
[0699] Example 72 was prepared from Example 71 in a similar manner
as Example 30. Preparative reverse phase HPLC separation method:
Start % B=0, Final % B=100, Gradient time=6 min, Flow Rate=45
mL/min, Column: phenomenex-Luna 30.times.50 mm S5, Fraction
Collection: 4.51-4.92 min; .sup.1H NMR: (CD.sub.3OD) .delta. 8.83
(d, J=4.5, 1H), 8.56 (d, J=8, 1H), 8.48 (s, 1H), 8.43 (d, J=8.5,
1H), 8.38 (d, J=2, 1H), 8.10-8.00 (overlapping m, 3H), 7.90-7.86 (d
overlapping with m, 2H), 7.67 (d, J=7, 1H), 7.50 (app t, 1H), 4.18
(m, 2H), 4.03 (m, 2H), 3.96 (m, 4H); Analytical HPLC method: Start
% B=0, Final % B=100, Gradient time=2 min, Flow Rate=5 mL/min,
Column: Xterra MS C18 S7 30.0.times.50 mm; LC/MS: (ES+) m/z
(M+H).sup.+=481.23, HPLC R.sub.t=1.147.
EXAMPLE 73
[0700] 191
[0701] Example 73 was prepared in a similar manner as Example 53.
.sup.1H NMR: (CD.sub.3OD) .delta. 8.76 (s, 1H), 8.47 (s, 1H), 8.30
(d, J=8.5, 1H), 8.07 (app t overlapping with s, 2H), 7.85 (d, J=8,
1H), 7.79 (app t, 1H), 4.50 (m, 2H), 4.39 (m, 2H), 4.04 (m, 2H),
3.88 (m, 2H); Analytical HPLC method: Solvent A 5% MeCN-95%
H.sub.2O-10 mM NH.sub.4OAc; Solvent B 95% MeCN-5% H.sub.2O-10 mM
NH.sub.4OAc; Start % B=0, Final % B=100, Gradient time=2 min, Flow
Rate=5 mL/min, Column: phenomenex 5u 4.6.times.50 mm C18; LC/MS:
(ES-) m/z (M+H).sup.+=481,483, HPLC R.sub.t=1.11.
EXAMPLE 74
[0702] 192
[0703] Example 74 was prepared from Example 73 with nBu.sub.3SnCN
(Pd(PPh.sub.3).sub.4, 1,4-dioxane 135.degree. C.). Preparative
reverse phase HPLC separation method: Start % B=0, Final % B=100,
Gradient time=6 min, Flow Rate=30 mL/min, Column: Xterra
19.times.50 mm S5, Fraction Collection: 3.26-3.65 min; Analytical
HPLC method: Start % B=0, Final % B=100, Gradient time=2 min, Flow
Rate=5 mL/min, Column: Xterra 4.6.times.50 mm C18 5u; LC/MS: (ES+)
m/z (M+H).sup.+=429.99, HPLC R.sub.t=1.020.
EXAMPLE 75
[0704] 193
[0705] Example 75 was prepared in as similar as Example 53.
Preparative reverse phase HPLC separation method: Start % B=0,
Final % B=100, Gradient time=6 min, Flow Rate=30 mL/min, Column:
Xterra 19.times.50 mm S5, Fraction Collection: 3.43-4.03 min;
.sup.1H NMR: (CD.sub.3OD) .delta. 8.76 (s, 1H), 8.37 (s, 1H), 8.31
(d, J=8, 1H), 8.08 (app t, 1H), 7.85 (d, J=8, 1H), 7.79 (app t
overlapping with s, 2H), 4.53 (m, 2H), 4.41 (m, 2H), 4.05-4.03 (m,
2H), 4.03 (s, 3H), 3.84 (m, 2H); Analytical HPLC method: Solvent A
5% MeCN-95% H.sub.2O-10 mM NH.sub.4OAc; Solvent B 95% MeCN-5%
H.sub.2O-10 mM NH.sub.4OAc; Start % B=0, Final % B=100, Gradient
time=2 min, Flow Rate=5 mL/min, Column: phenomenex 5u 4.6.times.50
mm C18; LC/MS: (ES-) m/z (M+H).sup.+=449.16, 451.21, HPLC
R.sub.t=1.033.
EXAMPLE 76
[0706] 194
[0707] Example 76 was prepared in a similar manner as Example 73.
Analytical HPLC method: Solvent A 5% MeCN-95% H.sub.2O-10 mM
NH.sub.4OAc; Solvent B 95% MeCN-5% H.sub.2O-10 mM NH.sub.4OAc;
Start % B=0, Final % B=100, Gradient time=2 min, Flow Rate=5
mL/min, Column: phenomenex 5u 4.6.times.50 mm C18; LC/MS: (ES-) m/z
(M+H).sup.+=481, 483, HPLC R.sub.t=1.108.
EXAMPLE 77
[0708] 195
[0709] Example 77 was prepared from Example 76 in a similar manner
as Example 72. Preparative reverse phase HPLC separation method:
Start % B=0, Final % B=100, Gradient time=10 min, Flow Rate=30
mL/min, Column: Xterra 19.times.50 mm S5, Fraction Collection:
5.47-5.72 min; .sup.1H NMR: (CD.sub.3OD) .delta. 9.39 (s, 1H), 8.82
(b s, 1H), 8.71-8.70 (m, 1H), 8.59-8.56 (m, 1H), 8.45 (d, J=9.2,
1H), 8.36 (dd, 1H), 8.27-8.25 (m, 1H), 8.15-8.09 (m, 1H), 8.03-7.99
(m, 1H), 7.53 (s, 1H), 7.50-7.47 (m, 1H), 4.12 (m, 2H), 3.90 (m,
2H), 3.85 (m, 2H), 3.74 (m, 2H); Analytical HPLC method: Start %
B=0, Final % B=100, Gradient time=2 min, Flow Rate=5 mL/min,
Column: phenomenex C18 3.0.times.50 mm 10 u, HPLC
R.sub.t=1.417.
EXAMPLE 78
[0710] 196
[0711] Example 78 was prepared from Example 76 in a similar manner
as Example 77. Preparative reverse phase HPLC separation method:
Start % B=0, Final % B=100, Gradient time=6 min, Flow Rate=30
mL/min, Column: Xterra 19.times.50 mm S5, Fraction Collection:
3.06-3.48 min; .sup.1H NMR: (CD.sub.3OD) .delta. 9.82 (s, 1H), 9.52
(s, 1H), 8.83 (b s, 1H), 8.70 (m, 1H), 8.64 (s, 1H), 8.48 (m, 1H),
8.42 (m, 1H), 8.39 (d, J=8.6, 1H), 8.11 (d, 1H), 7.79 (m, 1H), 4.13
(m, 2H), 3.92-3.96 (m, 2H), 3.86 (m, 2H), 3.61 (m, 2H); Analytical
HPLC method: Solvent A 5% MeCN-95% H.sub.2O-10 mM NH.sub.4OAc;
Solvent B 95% MeCN-5% H.sub.2O-10 mM NH.sub.4OAc; Start % B=0,
Final % B=100, Gradient time=2 min, Flow Rate=5 mL/min, Column:
phenomenex 5u 4.6.times.50 mm C18; LC/MS: (ES+) m/z
(M+H).sup.+=483.08, HPLC R.sub.t=1.142.
EXAMPLE 79
[0712] 197
[0713] Example 79 was prepared from Example 73 in a similar manner
as Example 77. Preparative reverse phase HPLC separation method:
Start % B=0, Final % B=100, Gradient time=14 min, Flow Rate=30
mL/min, Column: Xterra 19.times.50 mm S5, Fraction Collection:
4.00-4.37 min; .sup.1H NMR: (CD.sub.3OD) .delta. 8.87-8.84 (b m,
2H), 8.77 (s, 1H), 8.50 (s, 1H), 8.43 (d, J=2.5, 1H), 8.32 (d,
J=10, 1H), 8.09 (overlapping m, 3H), 7.85 (d, J=10, 1H), 7.81 (app
t, 1H), 4.54-4.51 (m, 2H), 4.43-4.40 (m, 2H), 4.08-4.05 (m, 2H),
3.93-3.90 (m, 2H); Analytical HPLC method: Solvent A 5% MeCN-95%
H.sub.2O-10 mM NH.sub.4OAc; Solvent B 95% MeCN-5% H.sub.2O-10 mM
NH.sub.4OAc; Start % B=0, Final % B=100, Gradient time=2 min, Flow
Rate=5 mL/min, Column: phenomenex 5u 4.6.times.50 mm C18; LC/MS:
(ES-) m/z (M+H).sup.+=480.16, HPLC R.sub.t=1.05.
EXAMPLE 80
[0714] 198
[0715] Example 80 was prepared from Example 53 and
pyrimidine-5-boronic acid (Pd(PPh.sub.3).sub.4, K.sub.2CO.sub.3,
2:1 DMF/H.sub.2O, 135.degree. C.). Preparative reverse phase HPLC
separation method: Start % B=0, Final % B=100, Gradient time=14
min, Flow Rate=30 mL/min, Column: Xterra 19.times.50 mm S5,
Fraction Collection: 3.33-3.39 min; .sup.1H NMR: (300 MHz,
CD.sub.3OD) .delta. 9.34 (s,1H), 9.25, (s, 2H), 8.77 (s, 1H), 8.49
(s, 1H), 8.34 (d, J=8.4, 1H), 8.24 (s, 1H), 8.09 (app t, 1H), 7.85
(d, J=9, 1H), 7.81 (app t, 1H), 4.57-4.53 (m, 2H), 4.46-4.42 (m,
2H), 4.14 (s, 3H), 4.09-4.05 (m, 2H), 3.91-3.87 (m, 2H); Analytical
HPLC method: Solvent A 5% MeCN-95% H.sub.2O-10 mM NH.sub.4OAc;
Solvent B 95% MeCN-5% H.sub.2O-10 mM NH.sub.4OAc; Start % B=0,
Final % B=100, Gradient time=2 min, Flow Rate=5 mL/min, Column:
phenomenex 5u 4.6.times.50 mm C18; LC/MS: (ES-) m/z
(M+H).sup.+=493.21, HPLC R.sub.t=0.957.
EXAMPLE 81
[0716] 199
[0717] Example 81 was prepared from Example 73 in a similar manner
as Example 80. Preparative reverse phase HPLC separation method:
Start % B=0, Final % B=100, Gradient time=13 min, Flow Rate=30
mL/min, Column: Xterra 19.times.50 mm S5, Fraction Collection:
4.28-4.89 min; .sup.1H NMR: (CD.sub.3OD) .delta. 9.34 (s,1H), 9.27,
(s, 2H), 8.77 (s, 1H), 8.51 (s, 1H), 8.43 (d, J=2.5, 1H), 8.32 (d,
J=8.5, 1H), 8.08 (app t, 1H), 7.86 (d, J=8.5, 1H), 7.81 (app t,
1H), 4.55-4.53 (m, 2H), 4.44-4.42 (m, 2H), 4.07-4.05 (m, 2H),
3.92-3.90 (m, 2H); Analytical HPLC method: Solvent A 5% MeCN-95%
H.sub.2O-10 mM NH.sub.4OAc; Solvent B 95% MeCN-5% H.sub.2O-10 mM
NH.sub.4OAc; Start % B=0, Final % B=100, Gradient time=2 min, Flow
Rate=5 mL/min, Column: phenomenex 5u 4.6.times.50 mm C18; LC/MS:
(ES+) m/z (M+H).sup.+=483.16, HPLC R.sub.t=0.997.
EXAMPLE 82
[0718] 200
[0719] Example 82 was prepared from Example 73 in a similar manner
as Example 81. Preparative reverse phase HPLC separation method:
Start % B=0, Final % B=100, Gradient time=12 min, Flow
Rate=30mL/min, Column: Xterra 19.times.50 mm S5, Fraction
Collection: 4.69-5.30 min; .sup.1H NMR: (CD.sub.3OD) .delta. 9.03
(s, 2H), 8.77 (s, 1H), 8.53 (s, 1H), 8.40 (d, J=3, 1H), 8.32 (d,
J=8.5, 1H), 8.08 (app t, 1H), 7.86 (d, J=8, 1H), 7.80 (app t, 1H),
4.55-4.53 (m, 2H), 4.44-4.42 (m, 2H), 4.15 (s, 3H), 4.07-4.05 (m,
2H), 3.92-3.90 (m, 2H); Analytical HPLC method: Solvent A 5%
MeCN-95% H.sub.2O-10 mM NH.sub.4OAc; Solvent B 95% MeCN-5%
H.sub.2O-10 mM NH.sub.4OAc; Start % B=0, Final % B=100, Gradient
time=2 min, Flow Rate=5 mL/min, Column: phenomenex 5u 4.6.times.50
mm C18; LC/MS: (ES-) m/z (M+H).sup.+=511.27, HPLC
R.sub.t=1.070.
EXAMPLE 83
[0720] 201
[0721] Example 83 was prepared from Example 73 in a similar manner
as Example 81. Preparative reverse phase HPLC separation method:
Start % B=0, Final % B=100, Gradient time=13 min, Flow Rate=30
mL/min, Column: Xterra 19.times.50 mm S5, Fraction Collection:
3.24-3.85 min; .sup.1H NMR: (CD.sub.3OD) .delta. 9.31 (s, 1H), 8.96
(d, J=5, 1H), 8.92 (d, J=5, 1H), 8.77 (s, 1H), 8.56 (s, 1H), 8.46
(d, J=2.5, 1H), 8.32 (d, J=8, 1H), 8.15 (dd, J=8, 5.5, 1H), 8.09
(app t, 1H), 7.87 (d, J=5, 1H), 7.81 (app t, 1H), 4.55-4.53 (m,
2H), 4.45-4.43 (m, 2H), 4.08-4.05 (m, 2H), 3.93-3.91 (m, 2H);
Analytical HPLC method: Solvent A 5% MeCN-95% H.sub.2O-10 mM
NH.sub.4OAc; Solvent B 95% MeCN-5% H.sub.2O-10 mM NH.sub.4OAc;
Start % B=0, Final % B=100, Gradient time=2 min, Flow Rate=5
mL/min, Column: phenomenex 5u 4.6.times.50 mm C18; LC/MS: (ES-) m/z
(M+H).sup.+=480.26, HPLC R.sub.t=1.033.
EXAMPLE 84
[0722] 202
[0723] Example 84 was prepared from Example 73 in a similar manner
as Example 72. Preparative reverse phase HPLC separation method:
Start % B=0, Final % B=100, Gradient time=10 min, Flow Rate=30
mL/min, Column: Xterra 19.times.50 mm S5, Fraction Collection:
5.40-5.55 min; Analytical HPLC method: Solvent A 5% MeCN-95%
H.sub.2O-10 mM NH.sub.4OAc; Solvent B 95% MeCN-5% H.sub.2O-10 mM
NH.sub.4OAc; Start % B=0, Final % B=100, Gradient time=2 min, Flow
Rate=5 mL/min, Column: phenomenex 5u 4.6.times.50 mm C18; LC/MS:
(ES+) m/z (M+H).sup.+=483.48, HPLC R.sub.t=1.048.
EXAMPLE 85
[0724] 203
[0725] As for the preparation of Example 39, Example 85 was
prepared in a similar manner as Example 36 using
4-piperazinylquinoline, which was prepared from the coupling of
4-chloroquinoline with tert-butyl 1-piperazinecarboxylate (CuBr,
Cs.sub.2CO.sub.3, DMF, sealed tube, 150.degree. C.) followed by
deprotection (HCl, 1,4-dioxane, r.t.). Preparative reverse phase
HPLC separation method: Start % B=0, Final % B=100, Gradient time=6
min, Flow Rate=45 mL/min, Column: Xterra MS C18 5 um 30.times.50
mm, Fraction Collection: 3.49-3.89 min; .sup.1H NMR: (CD.sub.3OD)
.delta. 9.37 (s,1H), 8.62 (d, J=6, 1H), 8.36 (s, 1H), 8.34 (app d,
1H), 8.27 (d, J=8.5, 1H), 7.98 (d, J=8.5, 1H), 7.92 (s, 1H),
7.93-7.92 (b m, 1H), 7.71 (app t, 1H), 7.23 (d, J=6.5, 1H), 4.10 (s
overlapping with m, 5H), 3.89-3.86 (m, 6H); Analytical HPLC method:
Start % B=0, Final % B=100, Gradient time=2 min, Flow Rate=5
mL/min, Column: Xterra MS C18 S7 30.0.times.50 mm; LC/MS: (ES+) m/z
(M+H).sup.+=483.18, HPLC R.sub.t=0.893.
EXAMPLE 86
[0726] 204
[0727] Example 86 and the corresponding
2-methoxy-6-methyl-4-piperazinylqu- inoline were prepared in a
similar manner as those described for Example 85. The crude
material was purified by preparative TLC (10%
MeOH/CH.sub.2Cl.sub.2) to give a white solid. Analytical HPLC
method: Start % B=0, Final % B=100, Gradient time=2 min, Flow
Rate=5 mL/min, Column: Xterra MS C18 S7 3.0.times.50 mm; LC/MS:
(ES+) m/z (M+H).sup.+=527.31, HPLC R.sub.t=1.100.
EXAMPLE 87
[0728] 205
[0729] Example 87 was prepared from Example 71 in a similar manner
as Example 74. Preparative reverse phase HPLC separation method:
Start % B=0, Final % B=100, Gradient time=6 min, Flow Rate=45
mL/min, Column: phenomenex-Luna 30.times.50 mm S5, Fraction
Collection: 3.73-4.06 min; .sup.1H NMR: (CD.sub.3OD) 8.60 (s, 1H),
8.45-8.39 (overlapping m, 2H), 8.09 (app d, 1H), 8.04 (app t, 1H),
7.88 (d, J=6.5, 1H), 7.87 (m, overlapping with d, 1H), 7.66 (d,
J=6.5, 1H), 4.18-4,14 (m, 2H), 4.03-3.99 (m, 2H), 3.96-3.91 (m,
4H); Analytical HPLC method: Start % B=0, Final % B=100, Gradient
time=2 min, Flow Rate=5 mL/min, Column: Xterra MS C18 S7
3.0.times.50 mm; LC/MS: (ES+) m/z (M+H).sup.+=429.13, HPLC
R.sub.t=0.933.
EXAMPLE 88
[0730] 206
[0731] Example 88 was prepared from Example 87 and benzoic
hydrazide by stirring in .sup.nBuOH in the presence of
K.sub.2CO.sub.3 in an sealed tube at 150.degree. C. for 2 h. After
evaporation of the volatile, the crude mixture was diluted with
methanol and purified by preparative reverse phase HPLC; Separation
method: Start % B=0, Final % B=100, Gradient time=6 min, Flow
Rate=45 mL/min, Column: phenomenex-Luna 30.times.50 mm S5, Fraction
Collection: 4.64-4.95 min; .sup.1H NMR: (CD.sub.3OD) 8.56 (s, 1H),
8.42 (d, J=8, 1H), 8.28 (d, J=7.5, 1H), 8.08 (app d, 1H), 8.02 (app
t, 1H), 7.89 (d, J=6.5, 1H), 7.86 (m overlapping with d, 2H), 7.65
(d, J=6.5, 1H), 7.59-7.54 (m, 4H), 4.19-4,17 (m, 2H), 4.01-3.97 (m,
4H), 3.93-3.89 (m, 2H); Analytical HPLC method: Solvent A 5%
MeCN-95% H.sub.2O-10 mM NH.sub.4OAc; Solvent B 95% MeCN-5%
H.sub.2O-10 mM NH.sub.4OAc; Start % B=0, Final % B=100, Gradient
time=2 min, Flow Rate=5 mL/min, Column: Phenomenex Luna C18 5 um
30.0.times.50 mm; LC/MS: (ES-) m/z (M+H).sup.+=545.16, HPLC
R.sub.t=1.343.
EXAMPLE 89
[0732] 207
[0733] Example 89 was isolated from the crude mixture of the
reaction to prepare Example 88. Preparative reverse phase HPLC
separation method: Start % B=0, Final % B=100, Gradient time=6 min,
Flow Rate=45 mL/min, Column: phenomenex-Luna 30.times.50 mm S5,
Fraction Collection: 4.36-4.63 min; .sup.1H NMR: (CD.sub.3OD) 8.52
(s, 1H), 8.43 (d, J=8.5, 1H), 8.36 (d, J=2.5, 1H), 8.09 (app d,
1H), 8.04 (app t, 1H), 8.01 (d, J=7.5, 2H), 7.88 (m overlapping
with d, 1H), 7.87 (d, J=6.5, 1H), 7.66 (d, J=6.5, 1H), 7.64 (app t,
1H), 7.58 (d, J=7.5, 1H), 7.57 (app t, 1H), 4.19-4,17 (m, 2H),
4.03-4.01 (m, 2H), 3.98-3.90 (m, 4H); Analytical HPLC method: Start
% B=0, Final % B=100, Gradient time=2 min, Flow Rate=5 mL/min,
Column: Xterra MS C18 5 um 30.0.times.50 mm; LC/MS: (ES+) m/z
(M+H).sup.+=565.17, HPLC R.sub.t=1.137.
[0734] Biology
[0735] ".mu.M" means micromolar;
[0736] "mL" means milliliter;
[0737] ".mu.l" means microliter;
[0738] "mg" means milligram;
[0739] The materials and experimental procedures used to obtain the
results reported in Tables 1-2 are described below.
[0740] Cells:
[0741] Virus production-Human embryonic Kidney cell line, 293, was
propagated in Dulbecco's Modified Eagle Medium (Invitrogen,
Carlsbad, Calif.) containing 10% fetal Bovine serum (FBS, Sigma,
St. Louis, Mo.).
[0742] Virus infection--Human epithelial cell line, HeLa,
expressing the HIV-1 receptor CD4 was propagated in Dulbecco's
Modified Eagle Medium (Invitrogen, Carlsbad, Calif.) containing 10%
fetal Bovine serum (FBS, Sigma, St. Louis, Mo.) and supplemented
with 0.2 mg/mL Geneticin (Invitrogen, Carlsbad, Calif.).
[0743] Virus-Single-round infectious 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 et al, Ref. 41).
Transfections were performed using lipofectAMINE PLUS reagent as
described by the manufacturer (Invitrogen, Carlsbad, Calif.).
[0744] Experiment
[0745] 1. HeLa CD4 cells were plated in 96 well plates at a cell
density of 1.times.10.sup.4 cells per well in 100 t Dulbecco's
Modified Eagle Medium containing 10% fetal Bovine serum and
incubated overnight.
[0746] 2. Compound was added in a 2 .mu.l dimethylsulfoxide
solution, so that the final assay concentration would be .ltoreq.10
.mu.M.
[0747] 3. 100 .mu.l of single-round infectious reporter virus in
Dulbecco's Modified Eagle Medium was then added to the plated cells
and compound at an approximate multiplicity of infection (MOI) of
0.01, resulting in a final volume of 200 III per well.
[0748] 4. Virally-infected cells were incubated at 37 degrees
Celsius, in a CO.sub.2 incubator, and harvested 72 h after
infection.
[0749] 5. Viral infection was monitored by measuring luciferase
expression from viral DNA in the infected cells using a luciferase
reporter gene assay kit, as described by the manufacturer (Roche
Molecular Biochemicals, Indianapolis, Ind.). Infected cell
supernatants were removed and 50 .mu.l of lysis buffer was added
per well. After 15 minutes, 50 g of freshly-reconstituted
luciferase assay reagent was added per well. Luciferase activity
was then quantified by measuring luminescence using a Wallac
microbeta scintillation counter.
[0750] 6. 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.
[0751] 7. An EC.sub.50 provides a method for comparing the
antiviral potency of the compounds of this invention. The effective
concentration for fifty percent inhibition (EC.sub.50) was
calculated with the Microsoft Excel xlfit curve fitting software.
For each compound, curves were generated from percent inhibition
calculated at 10 different concentrations by using a four
paramenter logistic model (model 205). The EC.sub.50 data for the
compounds is shown in Table 2. Table 1 is the key for the data in
Table 2.
[0752] Results
8TABLE 1 Biological Data Key for EC.sub.50s Compounds* with
Compounds with Compounds with EC.sub.50s > 5.mu.M EC.sub.50s
> 1 .mu.M but < 5 .mu.M EC.sub.50 < 1 .mu.M Group C Group
B Group A
[0753] *Some of these compounds may have been tested at a
concentration lower than their EC.sub.50 but showed some ability to
cause inhibition and thus should be evaluated at a higher
concentration to determine the exact EC.sub.50.
[0754] In Table 2, X.sub.w, X.sub.z and X.sub.a indicates the point
of attachment.
9TABLE 2 Examples 208 EC.sub.50 Table Entry Group (Example from
Number.) Z W A Table 1 1 (Example 1) 209 210 211 A 2 (Example 2)
212 213 214 A 3 (Example 3) 215 216 217 A 4 (Example 4) 218 219 220
A 5 (Example 5) 221 222 223 A 6 (Example 6) 224 225 226 A 7
(Example 7) 227 228 229 A 8 (Example 8) 230 231 232 A 9 (Example 9)
233 234 235 A 10 (Example 10) 236 237 238 A 11 (Example 11) 239 240
241 A 12 (Example 12) 242 243 244 A 13 (Example 13) 245 246 247 A
14 (Example 14) 248 249 250 A 15 (Example 15) 251 252 253 A 16
(Example 16) 254 255 256 A 17 (Example 17) 257 258 259 A 18
(Example 18) 260 261 262 A 19 (Example 19) 263 264 265 A 20
(Example 20) 266 267 268 A 21 (Example 21) 269 270 271 A 22
(Example 22) 272 273 274 A 23 (Example 23) 275 276 277 A 24
(Example 24) 278 279 280 A 25 (Example 25) 281 282 283 A 26
(Example 26) 284 285 286 A 27 (Example 27) 287 288 289 A 28
(Example 28) 290 291 292 A 29 (Example 29) 293 294 295 A 30
(Example 30) 296 297 298 A 31 (Example 31) 299 300 301 A 32
(Example 32) 302 303 304 A 33 (Example 33) 305 306 307 A 34
(Example 34) 308 309 310 A 35 (Example 35) 311 312 313 A 36
(Example 36) 314 315 316 A 37 (Example 37) 317 318 319 A 38
(Example 38) 320 321 322 A 39 (Example 39) 323 324 325 A 40
(Example 40) 326 327 328 A 41 (Example 41) 329 330 331 B 42
(Example 42) 332 333 334 A 43 (Example 43) 335 336 337 A 44
(Example 44) 338 339 340 A 45 (Example 45) 341 342 343 A 46
(Example 46) 344 345 346 A 47 (Example 47) 347 348 349 A 48
(Example 48) 350 351 352 A 49 (Example 49) 353 354 355 A 50
(Example 50) 356 357 358 A 51 (Example 51) 359 360 361 A 52
(Example 52) 362 363 364 A 53 (Example 53) 365 366 367 A 54
(Example 54) 368 369 370 A 55 (Example 55) 371 372 373 A 56
(Example 56) 374 375 376 A 57 (Example 57) 377 378 379 A 58
(Example 58) 380 381 382 A 59 (Example 59) 383 384 385 A 60
(Example 60) 386 387 388 A 61 (Example 61) 389 390 391 A 62
(Example 62) 392 393 394 A 63 (Example 64) 395 396 397 A 64
(Example 65) 398 399 400 A 65 (Example 66) 401 402 403 A 66
(Example 67) 404 405 406 A 67 (Example 68) 407 408 409 A 68
(Example 69) 410 411 412 A 69 (Example 70) 413 414 415 A 70
(Example 72) 416 417 418 A 71 (Example 73) 419 420 421 C 72
(Example 75) 422 423 424 A 73 (Example 77) 425 426 427 A 74
(Example 78) 428 429 430 A 75 (Example 79) 431 432 433 on test 76
(Example 80) 434 435 436 on test 77 (Example 81) 437 438 439 on
test 78 (Example 82) 440 441 442 on test 79 (Example 83) 443 444
445 on test 80 (Example 84) 446 447 448 on test 81 (Example 85) 449
450 451 on test A 82 (Example 86) 452 453 454 on test A 83 (Example
87) 455 456 457 on test 84 (Example 88) 458 459 460 on test 85
(Example 89) 461 462 463 on test
[0755] Guide to reading the structures shown in Table 2 above. The
structure of example 1 in the table above is: 464
[0756] 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 diluents.
[0757] 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.
[0758] 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.
[0759] 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.
[0760] 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.
[0761] 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 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.
* * * * *