Fab i inhibitors

(12) PATENT DESCRIPTION (19) PL (11) 201627 (13) B1 (21) Filing number: 354892 (22) Filing date: 10/06/2000 (86) Date and number of the international application:

06.10.2000, PCT / US00 / 27844 (87) International application publication date and number:

2001-04-19, WO01 / 27103 PCT Gazette No. 16/01 (51) Int.Cl.

C07D 401/12 (2006.01) C07D 405/12 (2006.01) C07D 471/04 (2006.01) C07D 487/04 (2006.01) C07D 495/04 (2006.01) A61K 31/381 (2006.01) A61K 31/47 (2006.01) A61P 31/00 (2006.01)

A new amide derivative, a pharmaceutical composition containing it and its uses (73) The patentee:

Affinium Pharmaceuticals, Inc., Toronto, CA (30) Priority:

10/08/1999, US, 60 / 158,704 (43) Application announced:

22.03.2004 BUP 06/04 (45) The following was announced about the grant of the patent:

30.04.2009 WUP 04/09 (72) Inventor (s):

William H. Miller, Collegeville, US Kenneth A. Newlander, West Chester, US Mark A. Seefeld, Collegeville, US

Irene N. Uzinskas, Villanova, US Walter E. Jr DeWolf, Glenmoore, US Dalia R. Jakas, Norristown, US (74) Agent:

Hawrylak Jolanta, PATPOL Sp. z o. o

⁽⁵⁷⁾ The present invention relates to a novel compound of formula (I) in which the substituents have the meaning as described herein, the composition containing the same and its uses. The compound of the invention of formula (I) is a pharmaceutically active compound and it inhibits Fab I and is useful for the treatment of bacterial infections.

<img file="PL201627B1_D0001.tif" />

PL 201 627 B1

Description of the invention

The present invention relates to a novel compound, a composition containing the same and to its uses. The compound of the invention is pharmaceutically active and inhibits Fab I and is useful for the treatment of bacterial infections.

While the general pathway of saturated fatty acid biosynthesis is similar in all organisms, the fatty acid synthase (FAS) systems differ significantly in their structural organization. Vertebrates and yeasts have FAS in which all enzymatic activities are encoded in one or two polypeptide chains, respectively, and the acyl carrier protein (ACP) is an integral part of the complex. In contrast, in bacterial FAS, each reaction is catalyzed by a separate monofunctional enzyme and ACP is a separate protein. There is therefore a great possibility of selectively inhibiting the bacterial system with antibacterial agents.

Fab I (previously designated EnvM) functions as enoyl-ACP reductase (Bergler et al. (1994), J. Biol. Chem. 269, 5493-5496) in the final step of the four reactions contained in each cycle of bacterial fatty acid biosynthesis. The first step in this pathway is catalyzed by β-ketoacyl-ACP synthase, which condenses malonyl-ACP with acetyl-CoA (Fab H, synthase III). In the following rounds, malonyl-ACP condenses with the growing acyl-ACP chain (Fab B and Fab F, synthases I and II, respectively). The second step in the extension cycle is ketoester reduction by NADPH dependent β-ketoacyl-ACP reductase (Fab G).

Subsequent dehydration by β-hydroxyacyl-ACP dehydrase (either Fab A or Fab Z) leads to trans-2-enoyl-ACP, which in turn is converted to acyl-ACP by enoyl-ACP reductase (Fab I) NADH dependent. Further rounds of this cycle, the addition of two carbon atoms per cycle, ultimately lead to palmitoyl-ACP (16C), where the cycle is largely halted due to the Fab I feedback inhibition by palmitoyl-ACP (Heath et al., (1996), J. Biol. Chem. 271, 1833-1836). Thus, Fab I is the major biosynthetic enzyme and is a key regulatory point in the entire synthetic pathway of bacterial fatty acid biosynthesis. Thus, Fab I is an ideal target for antimicrobial intervention.

Studies have shown that diazaborane antibiotics inhibit the biosynthesis of fatty acids, phospholipids and lipopolysaccharides (LPS) and that the antibacterial target of these compounds is Fab I. For example, the 2bl8 derivative known from Grassberger et al. (1984) J. Med. Chem. 27, 947-953 has been reported to be a non-competitive Fab I inhibitor (Bergler et al., (1994) J. Biol. Chem. 269, 5493-5496). In addition, plasmids containing the Fab I gene from a diazaborate resistant strain of S. typhimurium conferred resistance to diazaborane in E. coli (Turnowsky et al. (1989) J. Bacteriol. 171, 6555-6565). Moreover, inhibition of Fab I either by diazaborane or by elevation of temperature is lethal for the temperature sensitive Fab I mutant. These results show that Fab I is essential for the survival of the organism (Bergler et al. (1994) J. Biol. Chem. 269, 5493-5496).

Recent studies have shown that Fab I is also a target for triclosan, a broad-spectrum antibacterial agent (McMurry et al. (1998), Nature 394, 531-532). The crystal structure of E. coli Fab I complexed with NAD and triclosan shows that triclosan acts as an active site-directed, very potent inhibitor of Fab I by mimicking its natural substrate (Levy et al. (1999), Nature 398, 383-384) . Ward et al. ((1999), Biochem. 38, 12514-12525) showed that there is no evidence of formation of a covalent complex between Fab I and triclosan that would be analogous to the diazaboranes; Triclosan differs from these compounds in that it is a reversible Fab I inhibitor. The structural data for the Fab I complex of NAD and triclosan provides important information about Fab I as a therapeutic target.

Importantly, it has now been found that certain compounds are Fab I inhibitors and have antimicrobial activity, and therefore may be useful for the treatment of bacterial infections in mammals, particularly humans.

Additionally, two of the present Fab I inhibitory compounds have been found to be inhibitors of Streptococcus Fab K.

Fab I is absent in Streptococcus, and Fab I is not essential in Pseudomonas. There is also reason to believe that Fab I may not be essential in Enterococcus.

In all of these organisms, another enoyloreductase, called Fab K, is present (Heath, RJ; Rock, CO, Nature (2000), 406, 145-146). Pseudomonas and Enterococcus contain both Fab I,

PL 201 627 B1 and Fab K, and Streptococcus contains only Fab K. Consequently, it is not expected that pure Fab I inhibitors will have antimicrobial activity in these organisms.

Thus, compounds that inhibit both Fab I and Fab K have the potential to be broad-spectrum antimicrobial agents.

The present invention provides compounds of formula (I) as described hereinafter which inhibit Fab I and are useful in the treatment of bacterial infections.

The present invention provides compounds of formula (I) as described hereinafter which inhibit Fab I and are useful in the treatment of bacterial infections.

The present invention also provides a pharmaceutical composition comprising a compound of formula (I) and a pharmaceutically acceptable carrier.

The invention also relates to the use of a compound of formula (I) as a medicament and in the manufacture of a medicament for the treatment of bacterial infections, diseases for which inhibition of Fab I is indicated.

Detailed description

The present invention relates to a compound of formula (I):

<img file="PL201627B1_D0002.tif" />

wherein:

<img file="PL201627B1_D0003.tif" />

₁

R¹ is H or methyl;

₂

R^{2 3} is H, methyl, propyl or cyclopropyl;

R means

PL 201 627 B1

<img file="PL201627B1_D0004.tif" />

PL 201 627 B1

R⁶ is H or methyl;

<img file="PL201627B1_D0005.tif" />

means that one of the two marked bonds is a double bond and the other is a single bond;

X is H, F, methoxy or benzyl; or a pharmaceutically acceptable salt thereof.

The present invention also includes the pharmaceutically acceptable addition salts and complexes of the compounds of the present invention. In cases where the compounds of the present invention may have one or more chiral centers, unless specified, the present invention includes each unique racemic compound as well as each unique non-racemic compound.

In cases where the compounds have unsaturated carbon-carbon double bonds, both the cis (Z) and the trans (E) isomers are within the scope of the present invention. In cases where compounds can exist in tautomeric forms such as ketoenol tautomers such as OR '

Jk and -Λ, any tautomeric form is contemplated as being included in the present invention, whether in equilibrium or fixed in one embodiment by appropriate substitution with R '. The meaning of any substituent at any one occurrence is independent of its meaning or the meaning of any other substituent at any other occurrence.

The present invention also includes prodrugs of the compounds of the present invention. Prodrugs are any covalently linked carrier that releases the active parent drug of formula (I) in vivo.

Compounds of formula (I) inhibit Fab I. Inhibition of this enzyme is useful in the treatment of bacterial infections. The compounds of the present invention may also be useful as antifungal agents. Additionally, the compounds may be useful in combination with known antibiotics.

With respect to formula (I), the present invention preferably comprises compounds of formula (Ia):

<img file="PL201627B1_D0006.tif" />

in which R.², R³, R⁴, R⁵ and X are as defined above for compounds of formula (I). In a preferred embodiment, the invention relates to a compound of formula (II):

<img file="PL201627B1_D0007.tif" />

in which R.¹, R², R³ and X are as defined above for compounds of formula (I).

PL 201 627 B1

With respect to formula (II), the present invention preferably comprises compounds of formula (IIa):

<img file="PL201627B1_D0008.tif" />

in which R.¹, R², R³ and X are as defined above for compounds of formula (I).

In particular, as regards formula (II), the present invention preferably comprises compounds of formula (IIb):

<img file="PL201627B1_D0009.tif" />

in which R.³ is as defined above for compounds of formula (I). Conveniently, with respect to formula (I), R.³ means:

<img file="PL201627B1_D0010.tif" />

wherein X, Y, M, L and E are as defined for compounds of formula (I).

Representative of the novel compounds of the present invention are the compounds of the Examples

1-86 onwards.

Thus, in a preferred embodiment, the invention relates to a compound of formula (I) which is:

(E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -3- (4-Aminophenyl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (pyridin-3-yl) acrylic amide;

(E) -3- (2-Aminopyrimidin-5-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N- (benzo [b] thiophen-2-ylmethyl) -N-methylacrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -2-butenamide;

(E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indazol-3-ylmethyl) acrylamide;

(E) -3- (6-Amino-2-methylpyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -3- (6-Amino-5-methylpyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N- (1-methyl-1H-indol-2-ylmethyl) -N-propylacrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N- (5-fluoro-1-methyl-1H-indol-2-ylmethyl) -N-methylacrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (naphthalen-1-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -2, N-dimethyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (naphthalen-2-ylmethyl) acrylamide;

2- (6-Aminopyridin-3-ylmethyl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N- (benzofuran-2-ylmethyl) -N-methylacrylamide;

(E) -3- (3,4-Dihydro-2H-pyrido [3,2-b] -1,4-oxazin-7-yl) -N-methyl- (1-methyl-1H-indol-2- ilmethyl) acrylamide;

(E) -N-Methyl-3- [6- (methylamino) pyridin-3-yl] -N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -3- [6- (Dimethylamino) pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -N-Methyl-N - [(1-methyl-1H-indol-2-yl) methyl] -3- (5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl) acrylamide;

(E) -3- [6- (Acetylamino) pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -3- (6-Amino-5-methylpyridin-3-yl) -N- (benzo [b] thiophen-2-ylmethyl) -N-methylacrylamide;

(E) -3- (6-Amino-5-methylpyridin-3-yl) -N-methyl-N- (naphthalen-2-ylmethyl) acrylamide;

(E) -3- (6-Amino-4-methylpyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N-cyclopropyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-3-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (quinolin-3-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (thieno [2,3-b] thiophen-2-ylmethyl) acrylamide;

(E) -N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (6-methylpyridin-3-yl) acrylic amide;

(E) -3- [6- (Acetylamino) -5-methylpyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N- (1H-indol-2-ylmethyl) -N-methylacrylamide;

(E) -N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- [6- (2-oxopropylamino) pyridin-3-yl] acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (thieno [3,2-b] thiophen-2-ylmethyl) acrylamide;

(E) -3- [6-Amino-5- (hydroxymethyl) pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -acrylamide;

(E) -3- (3H-Imidazo [4,5-b] pyridin-6-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -3- [6-Aminopyridin-3-yl] -N- (1-ethyl-1H-indol-2-ylmethyl) -N-methylacrylamide;

(E) -3- [6-Aminopyridin-3-yl] -N- (1,3-dimethyl-1H-indol-2-ylmethyl) -N-methylacrylamide;

(E) -3- [6 - ((E) -But-2-enoylamino) pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl ) acrylamide;

(E) -3- [6-Amino-5 - [(2-hydroxyethylamino) carbonyl] pyridin-3-yl] -N- (1-methyl-1H-indol-2-ylmethyl) -N-methylacrylamide;

(E) -3- [6-Aminopyridin-3-yl] -N-methyl-N- (3-methyl-1H-inden-2-ylmethyl) acrylamide;

(E) -3- [6-Aminopyridin-3-yl] -N- (1H-inden-2-ylmethyl) -N-methylacrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (6-methyl-6H-thieno [2,3-b] pyrrol-5-ylmethyl) acrylamide;

(E) -N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (2-oxo-1,4-dihydro-2H-pyrido [2,3-d] -1, 3-oxazin-6-yl) acrylamide;

(E) -3- [6- (1,3-dioxo-1,3-dihydroisoindol-2-yl) pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-2- ilmethyl) acrylamide;

(E) -3- [6 - [(2-Carboxybenzoyl) amino] pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -acrylamide;

(E) -3- [6- (3-Ethylureido) pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -N- (1,3-Dimethyl-1H-indol-2-ylmethyl) -N-methyl-3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3 -yl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (3-methylbenzo [b] thiophen-2-ylmethyl) acrylamide; (E) -3- (6-Aminopyridin-3-yl) -N- (4-methoxy-1-methyl-1H-indol-2-ylmethyl) -N-methylacrylamide; (E) -3- [6- (Acetylamino) pyridin-3-yl] -N-methyl-N- (3-methyl-1H-inden-2-ylmethyl) acrylamide; (E) -3- [6- (Acetylamino) pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-3-ylmethyl) acrylamide;

(E) -N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (3-methyl-2-oxo-1,2,3,4-tetrahydropyrid- [2,3- d] -pyrimidin-6-yl) acrylamide;

(E) -N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- [6- (propionylamino) pyridin-3-yl] acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N- (1,4-dimethyl-1H-indol-2-ylmethyl) -N-methylacrylamide;

(E) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- [6- (3-methylureido) pyridin-3-yl] acrylic amide;

(E) -N-Methyl-N- (3-methyl-1H-inden-2-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl ) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (4-methyl-4H-thieno [3,2-b] pyrrol-5-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N- (3,4-dimethylthieno [2,3-b] thiophen-2-ylmethyl) -N-methylacrylamide;

(E) -N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- [6- (phenylamino) pyridin-3-yl] acrylic amide;

(E) -3- (6-Aminopyridin-3-yl) -N- (6-methoxy-1-methyl-1H-indol-2-ylmethyl) -N-methylacrylamide;

(E) -3 - (2-Aminopyrimidin-5-yl) -N- (benzo [b] thiophen-2-ylmethyl) -N-methylacrylamide;

(E) -3- (2-Aminopyrimidin-5-yl) -N-methyl-N- (1-methyl-1H-indol-3-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (1-methyl-naphthalen-2-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N- (1,2-dimethyl-1H-indol-3-ylmethyl) -N-methylacrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N- (benzo [b] thiophen-3-ylmethyl) -N-methylacrylamide;

(E) -N-Methyl-N- (1-methyl-1H-indol-3-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl ) acrylamide;

(E) -3- [2-Aminopyrimidin-5-yl] -N-methyl-N- (3-methyl-1H-inden-2-ylmethyl) acrylamide;

(E) -N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- [6- (pyridin-2-ylamino) pyridin-3-yl] acrylamide;

(E) -3- [2- (Acetylamino) pyrimidin-5-yl] -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (2-methyl-1H-indol-3-ylmethyl) acrylamide;

(E) -3- (2-Aminopyrimidin-5-yl) -N- (1,2-dimethyl-1H-indol-3-ylmethyl) -N-methylacrylamide;

(E) -N- (1,2-Dimethyl-1H-indol-3-ylmethyl) -N-methyl-3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3 -yl) acrylamide;

(E) -N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (3-oxo-3,4-dihydro-2H-pyrido [3,2-b] -1, 4-oxazin-7-yl) acrylamide;

(E) -N-Methyl-N- (3-methylbenzo [b] thiophen-2-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl ) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-pyrrolo [2,3-b] pyridin-3-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N- (1,7-dimethyl-1H-indol-3-ylmethyl) -N-methylacrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N- (1,5-dimethyl-1H-indol-3-ylmethyl) -N-methylacrylamide;

E) -N-Methyl-N- (2-methyl-1H-indol-3-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N- (1,6-dimethyl-1H-indol-3-ylmethyl) -N-methylacrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N- (2,3-dihydro-1H-3a-azacyclopenta [a] inden-8-yl) -N-methylacrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N-methyl- (2-methyl-benzo [b] thiophen-3-ylmethyl) acrylamide;

(E) -3- (6-Aminopyridin-3-yl) -N- (1-benzyl-1H-indol-3-ylmethyl) -N-methylacrylamide;

(E) -N-Methyl-N- (1-methyl-1H-indol-3-ylmethyl) -3- (3-oxo-3,4-dihydro-2H-pyrido [3,2-b] -1, 4-oxazin-7-yl) acrylamide; or (E) -N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3- ylo) propionamide; or a pharmaceutically acceptable salt thereof.

The compounds of the present invention are Fab I inhibitors useful in the treatment of bacterial infections. Two compounds of the present invention, namely (E) -N-methyl-N- (1-methyl-1H-indol-3-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1 , 8-naphthyridin-3-yl) acrylamide and (E) -N-methyl-N- (2-methyl-1H-indol-3-ylmethyl) -3- (7-oxo-5,6,7,8- tetrahydro-1,8-naphthyridin-3-yl) acrylamide, are dual Fab I / Fab K inhibitors. These compounds can be broad-spectrum antibiotics. Thus, the invention also relates to the use of (E) -N-methyl-N- (1-methyl-1H-indol-3-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8 -naphthyridin-3-yl) acrylamide or (E) -N-methyl-N- (2-methyl-1H-indol-3-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro 1,8-naphthyridin-3-yl) acrylamide for the manufacture of a medicament for the treatment of diseases for which inhibition of Fab K is indicated.

Abbreviations and symbols commonly used in chemistry, especially peptide chemistry, are used herein to describe the compounds of the invention. In general, the amino acid abbreviations follow the recommendations of The IUPAC-IUB Joint Commission on Biochemical Nomenclature as described in Eur. J. Biochem. 158,9 (1984).

PL 201 627 B1

The term C1-4alkyl as used herein denotes an optionally substituted alkyl group having 1 to 4 carbon atoms, and includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and t-butyl. The C1-6alkyl group further includes pentyl, n-pentyl, isopentyl, neopentyl and hexyl groups and simple aliphatic isomers thereof. The C0-4alkyl and C0-6alkyl groups further indicate that no alkyl group must be present (e.g. that a covalent bond is present).

Any C1-4alkyl or C1-6alkyl group may be optionally substituted with a R group^xwhich may be on any carbon atom that gives a stable structure and is accessible by conventional synthetic techniques.

The appropriate groups for R.^x are C1-4alkyl, OR ', SR', CN, N (R ') 2, CH2N (R') 2, -NO2, -CF3, -CO2R '-CON (R') 2, -COR ', - NR'C (O) R ', F, Cl, Br, I or -S (O) rCF3, where R' and r are as defined for compounds of formula (I).

Halogen or halogen is F, Cl, Br and I.

The term Ar as used herein or the aryl group means a phenyl or naphthyl group, or a phenyl or naphthyl group, substituted with one to three substituents as defined above for an alkyl group, or substituted with a methylenedioxy group.

Het or heterocycle is an optionally substituted five- or six-membered monocyclic ring or a nine- or ten-membered bicyclic ring containing one to three heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, which is stable and accessible by conventional chemical synthesis. Exemplary heterocycles are benzofuryl, benzimidazolyl, benzopyranyl, benzothienyl, furyl, imidazolyl, indolinyl, morpholinyl, piperidinyl, piperazinyl, pyrrolyl, pyrrolidinyl, tetrahydro-pyridinyl, pyridinyl, thiazolyl, perchinelinyl, quinolinyl, and isolinetinylinyl, isolinyl quinolinyl. It is within the scope of the present invention to make any available combination of no more than three substituents on the Het ring, such as defined above for an alkyl group, which is chemically synthesized and stable.

In the present description, abbreviations of the names of certain radical groups are used. t-Bu refers to tertiary butyl radical, Boc refers to t-butyloxycarbonyl radical, Fmoc refers to fluorenylmethoxycarbonyl radical, Ph refers to phenyl radical, Cbz refers to benzyloxycarbonyl radical, Bn refers to benzyl radical, Me refers to the methyl radical, Et refers to the ethyl radical, Ac refers to the acetyl radical, Alk refers to the C1-4alkyl group, Nph refers to the 1- or 2-naphthyl group, and cHex refers to the cyclohexyl group. Tet refers to the 5-tetrazolyl group.

In the present description, abbreviations of some reagents are used. DCC refers to dicyclohexylcarbodiimide, DMAP refers to dimethylaminopyridine, EDC refers to 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride, HOBt refers to 1-hydroxybenzotriazole, THF refers to tetrahydro-ADrofuran, DIEAropyl refers to diethyl azodicarboxylate, PPh3 refers to triphenylphosphine, DIAD refers to diisopropyl azodicarboxylate, DME refers to dimethoxyethane, DMF refers to dimethylformamide, NBS refers to N-bromosuccinimide, Pd / C refers to palladium on carbon catalyst, PPA refers to polyphosphoric acid, DPPA refers to diphenylphosphoryl azide, BOP refers to benzotriazol-1-yloxy-tris hexafluorophosphate (dimethylamino) phosphonium, HF refers to hydrofluoric acid, TEA refers to triethylamine, TFA refers to trifluoroacetic acid, PCC refers to pyridinium chlorochromate.

In general, the compounds of the present invention are prepared by:

(i) reacting a compound of formula (III) with a compound of formula (IV):

,4

NH

HO ^R (III) (IV) in which R.², R³, R⁴, R⁵ and x are as defined in formula (I), with all reactive functionalities protected, in the presence of EDC and HOBT;

(Ii) reacting a compound of formula (V) with a compound of formula (VI):

<img file="PL201627B1_D0011.tif" />

in which R.², R³ and x are as defined in formula (I), and halogen is Br, Cl, F or I, with all reactive functionalities protected, in the presence of a palladium (II) salt, a phosphine ligand and a base; followed by removing any protecting groups and optionally forming a pharmaceutically acceptable salt.

In particular, compounds of formula (I) are prepared by the general methods described in the Schemes hereinafter.

<img file="PL201627B1_D0012.tif" />

(a) benzyl acrylate, Pd (OAc) 2, P (o-tol) 3, (i-Pr) 2NEt, propionitrile;

(b) 1.0 N NaOH, MeOH;

(c) 1-methyl-2- (methylaminomethyl) indole, EDC, HOBt.H2O, Et3N, DMF.

A suitable haloaromatic derivative, e.g. 2-amino-5-bromopyridine (I-1), is reacted with the appropriate α, β-unsaturated ester, e.g. benzyl acrylate, in a Heck-type reaction (see Heck, Org. Reactions 1982, 27, 345). ) to form I-2. The reaction is palladium (0) mediated and is generally conducted in an inert solvent such as CHCN, propionitrile or toluene in the presence of a suitable acid removing reagent such as triethylamine (Et3N) or diisopropylethylamine ((i-Pr) 2NEt). Common sources of palladium (0) include palladium (II) acetate (Pd (OAc) 2) and palladium (II) chloride (PdCl2) and phosphine ligands are often used, for example triphenylphosphine (PPh3) or tri-ortho-tolylphosphine (P ( tol) 3). The ethyl ester I-2 is hydrolyzed using an aqueous base, for example LiOH in aqueous THF or NaOH in aqueous methanol or ethanol, and the intermediate carboxylate salt is acidified with a suitable acid, for example TFA or HCl, to give the carboxylic acid I-3. Carboxylic acid I-3 is converted to an activated form using, for example, EDC and HOBt or SOCl2, and the activated form is then reacted with a suitable amine, for example 1-methyl-2- (methylaminomethyl) indole, in a suitable solvent such as DMF , CH2Cl2 or CH3CN to give I-4. Depending on whether acid neutralization is needed, an added base such as triethylamine (Et3N), diisopropylethylamine ((i-Pr) 2NEt) or pyridine may be used.

There are many additional methods for converting carboxylic acid to amide, which can be found in standard references such as "Compendium of Organic Synthetic Methods", Volumes I-VI (published by Wiley-Interscience) or Bodansky, "The Practice of Peptide Synthesis" ( published by Springer-Verlag).

PL 201 627 B1

The term "amide coupling reagents" as used herein means reagents that can be used to form peptide bonds.

Typical coupling methods employ carbodiimides, activated anhydrides and esters, and acyl halides.

Reagents such as EDC, DCC, DPPA, PPA, BOP, HOBt, N-hydroxy succinimide and oxalyl chloride are typical.

Typically, the amine is coupled through its free amino group with the appropriate carboxylic acid using a suitable carbodiimide coupling agent such as N, N'-dicyclohexylcarbodiimide (DCC), optionally in the presence of catalysts such as 1-hydroxybenzotriazole (HOBt) and dimethylaminopyridine (DMAP) .

Other methods are also useful, such as forming activated esters, anhydrides or acid halides from the free carboxyl group of a suitably protected acid followed by reaction with the free amine, optionally in the presence of a base.

For example, benzoic acid is treated with isobutyl chloroformate in an anhydrous solvent such as methylene chloride or tetrahydrofuran (THF) in the presence of a base such as N-methylmorpholine, DMAP, or trialkylamine to form an "activated anhydride" which is then reacted with free amine.

<img file="PL201627B1_D0013.tif" />

(a) NaH, MeI, DMF;

(b) CH3NH2, H2O, MeOH;

(c) LiAlH4, THF.

The coupling amines used in the present invention were prepared by established methods known to those skilled in the art. For example, amine II-4 is prepared by the direct procedure outlined in Scheme II.

Commercially available ethyl indole-2-carboxylate (II-1) is deprotonated with an appropriate base, generally sodium hydride (NaH), and the intermediate sodium salt is reacted with a suitable alkylating agent, for example methyl iodide, to afford II-2.

Polar solvents such as DMF, THF or mixtures thereof are generally preferred for this reaction. Compound II-2 can conveniently be converted to II-3 by reaction with an excess of an amine such as methylamine in a polar solvent, generally H 2 O or a mixture of H 2 O and methanol.

Alternatively, the ester group II-2 can be saponified under standard conditions, typically using an alkali metal hydroxide such as LiOH, NaOH or KOH, in an aqueous solvent such as THF, ethanol or methanol and the resulting carboxylic acid can be converted to the desired amide.

Typical methods of amide formation are described in Scheme I. Reduction of amide II-3 to amine II-4 is typically done with lithium aluminum hydride (LiAlH4) in refluxing THF, although many other methods can be used to reduce amides to amines.

Such methods are known to those skilled in the art, and can be found in normal references such as "Compendium of Organic Synthetic Methods" (published by Wiley-Interscience).

PL 201 627 B1

<img file="PL201627B1_D0014.tif" />

(a) CH3NH2, NaCNBH3, MeOH.

The coupling amines used in the present invention can also be prepared by reductive amination of the corresponding aldehyde (Scheme III). This process, which is known to those skilled in the art, involves the initial conversion of the aldehyde into an intermediate imine, which in turn is reduced, often in situ, to give the amine. For example, a commercially available aldehyde III-1 is reacted with a suitable amine, e.g., methylamine, to form an intermediate imine (not shown) which is reduced in situ to amine III-2 by reaction with a suitable reducing agent, typically sodium cyanoborohydride or ( triacetoxy) sodium borohydride. Often the reaction is performed in the presence of an acid such as acetic acid in a polar solvent such as methanol or DMF.

<img file="PL201627B1_D0015.tif" />

(a) AC2O, NaHCO3, THF.

The amino group of compound IV-1 (prepared as described in Scheme I) reacts with a variety of acylating agents to form amides, sulfonamides, ureas, and carbamates. For example, IV-1 is reacted with acetic anhydride (AC2O) in an inert solvent, typically THF, in the presence of a suitable base such as sodium hydrogen carbonate (NaHCO3) to give IV-2. Other acylating agents, including sulfonyl halides, isocyanates, and chlorocarbonates, also participate in this reaction to form sulfonamides, ureas, and carbamates, respectively.

<img file="PL201627B1_D0016.tif" />

(a) H 2, Pd / C, EtOH;

(b) (Boc) 2O, LiHMDS, THF;

(c) NBS, AcOH, CH2Cl2;

(d) benzyl acrylate, Pd (OAc) 2, P (o-tol) 3, (i-Pr) 2NEt, propionitrile;

(e) 4 N HCl / dioxane;

(f) LiOH, H2O, MeOH.

PL 201 627 B1

1,8-Naphthyridine (V-1) can be selectively reduced to 1,2,3,4-tetrahydro-1,8-naphthyridine (V-2) by reaction with hydrogen gas in the presence of a suitable catalyst, preferably metal palladium on activated carbon (Pd / C), in an inert solvent, generally MeOH, EtOH, EtOAc, or mixtures thereof. V-2 is converted to a suitably protected derivative, for example an N-Boc-protected derivative of V-3, by reaction with di-tert-butyl dicarbonate in the presence of a suitable base, preferably lithium hexamethyldisilazide (LiHMDS). The amino protecting group must be compatible with the further chemical transformations and must be easily removable when desired. Methods for protecting amines are known to those skilled in the art, and are described in standard references such as Greene "Protective Groups in Organic Synthesis" (published by Wiley-Interscience). V-3 is selectively brominated at the 6-position by reaction with a suitable brominating agent such as bromine (Br2) or N-bromosuccinimide (NBS). Typical solvents for the bromination reaction include CH2Cl2, CCl4, MeOH, AcOH or mixtures thereof. The resulting V-4 6-bromo-1,2,3,4-tetrahydro-1,8-naphthyridine participates in the Heck reaction as described in scheme I to give V-5. Removal of the Boc protecting group is achieved under standard acidic conditions known to those skilled in the art (see Greene above) and the benzyl ester is saponified as described in scheme I to afford V-6.

Scheme VI

<img file="PL201627B1_D0017.tif" />

Η (a) LiAlH4, THF;

(b) NBS, CH2Cl2;

(c) 48% HBr;

(d) (MeO2C) 2CH2, NaOMe, MeOH;

(e) NaOH, H 2 O, MeOH, (f) HCl, H 2 O, MeOH;

(g) acryloyl chloride, EtN, CH2Cl2;

(h) Pd (OAc) 2, P (o-Tol) 3, (i-Pr) 2NEt, propionitrile.

PL 201 627 B1

Commercially available 2-aminonicotinic acid (VI-1) is reduced to alcohol VI-2 under standard conditions (LiAlH4, THF) and the aromatic ring of VI-2 is brominated using e.g. bromine or N-bromosuccinimide (NBS) in a solvent an inert such as CH2Cl2 to give VI-3. By reaction with 48% aqueous HBr, VI-3 is converted to bromide VI-4 which is reacted with a malonic acid diester, for example dimethyl malonate, in the presence of a suitable base, typically sodium methoxide, in an alcoholic solvent such as methanol with to form the naphthidone derivative VI-5. Saponification and neutralization under standard conditions gives an intermediate carboxylic acid (not shown) which is typically not isolated but decarboxylated with mild heating to yield the naphthyridone VI-6. This compound reacts with acrylamide VI-8 in a Heck-type reaction as described in scheme I to give VI-9. Alternatively, VI-6 can be converted to VI-9 following the general procedure described in Scheme I for converting I-1 to I-4. Acrylamide VI-8 will conveniently be formed by reacting the amine VI-7 (see Scheme II) with the activated form of acrylic acid by an amide bond formation reaction. Typical amide-forming conditions are described in Scheme I and are known to those skilled in the art.

<img file="PL201627B1_D0018.tif" />

(a) CH3NH2, H2O, THF;

(b) (MeO) 2C = O, NaOMe, MeOH;

(c) compound VI-8, Pd (OAc) 2, P (o-tol) 3, (i-Pr) 2NEt, propionitrile.

Benzyl bromide VII-1, prepared as described in scheme VI, reacts with an amine, for example aqueous methylamine, to give c benzylamine VII-2. Polar solvents such as THF, DMF, DMSO or a mixture thereof are generally preferred for this reaction. VII-2 is reacted with dialkyl carbonate, dimethyl carbonate, in the presence of a suitable base, typically sodium methoxide, in an alcoholic solvent, generally methanol, to give the cyclic urea derivative VII-3. This compound is converted to VII-4 by reaction with the compound VI-8 as described in scheme VI.

<img file="PL201627B1_D0019.tif" />

(A) SnCl2-H2O, EtOH;

(b) 96% HCO2H;

(c) TrCl, Et3N, CH2Cl2;

(d) benzyl acrylate, Pd (OAc) 2, P (o-tol) 3, (i-Pr) 2NEt, propionitrile;

(e) 4N HCl / dioxane;

(f) NaOH, H2O, MeOH.

The nitro group of commercially available 2-amino-5-bromo-3-nitropyridine (VIII-1) is reduced under standard conditions using for example tin (II) chloride in EtOH. The obtained diamine VIII-2 is reacted with formic acid or an appropriate equivalent to give the imidazopyridine derivative VIII-3. This compound is converted into a suitably protected derivative, for example the N-trityl protected derivative VIII-4, by reaction with trityl chloride in the presence of a suitable base, typically triethylamine or diisopropylethylamine. Typical solvents for this reaction include CH2Cl2, DMF or mixtures thereof. As shown in Scheme V, the amino protecting group must be compatible with the subsequent chemical transformations and must be easily removable when desired. VIII-4 is converted to VIII-6 following the general procedure in Scheme V.

<img file="PL201627B1_D0020.tif" />

(a) Br2, AcOH;

(b) N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide, Pd (OAc) 2, P (o-tol) 3, (i-Pr) 2NEt, propionitrile.

Commercially available 2,2'-dipyridylamine (IX-1) is monobromated at the 5-position by reaction with a suitable brominating agent such as bromine (Br2) or N-bromosuccinimide (NBS). Typical solvents for the bromination reaction include CH2Cl2, CCl4, MeOH, AcOH or mixtures thereof. The resulting monobromo derivative IX-2 is reacted with N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -acrylamide in a Heck-type reaction as described in Scheme I to give IX-3.

<img file="PL201627B1_D0021.tif" />

(a) Br2, AcOH;

(b) N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide, Pd (OAc) 2, P (o-tol) 3, (i-Pr) 2NEt, propionitrile.

PL 201 627 B1

Commercially available 2H-pyrido [3,2-b] -1,4-oxazin-3 (4H) -one (X-1) is selectively brominated at the 5-position by reaction with a suitable brominating agent such as bromine (Br2) or N-bromosuccinimide (NBS). Typical solvents for the bromination reaction include CH2Cl2, CCl4, MeOH, AcOH or mixtures thereof. The resulting monobromo derivative X-2 is reacted with N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide in a Heck type reaction as described in Scheme I to give X-3.

The acid addition salts of the compounds are prepared in the normal manner in a suitable solvent from the parent compound and an excess of an acid such as hydrochloric, hydrobromic, hydrofluoric, sulfuric, phosphoric, acetic, trifluoroacetic, maleic, succinic or methanesulfonic acid. Some of the compounds form inner salts or zwitterions which may be acceptable. Cationic salts are prepared by reacting the parent compound with an excess of an alkaline reagent such as hydroxide, carbonate or alkoxide having the appropriate cation; or with an appropriate organic amine. Cations such as Li are specific examples of cations present in pharmaceutically acceptable salts⁺, Na⁺, K⁺, Ca⁺⁺, Mg⁺⁺ and NH₄⁺.

The present invention also relates to a pharmaceutical composition containing one or more conventional excipients and, as an active ingredient, a compound of formula (I) as defined above. Accordingly, the compounds of formula (I) can be used in the preparation of a medicament. Pharmaceutical compositions of the compounds of formula (I) prepared as previously described may be prepared as solutions or lyophilized powders for parenteral administration. Before use, the drug may be reconstituted from the powder by adding an appropriate diluent or other pharmaceutically acceptable carrier. The liquid preparation can be a buffered, isotonic aqueous solution. Examples of suitable diluents are normal isotonic saline, normal 5% dextrose in water, or buffered sodium or ammonium acetate. Such a formulation is suitable especially for parenteral administration, but may also be used for oral administration or contained in an inhalation dispenser or insufflation pump. It may be desirable to add excipients such as polyvinylpyrrolidone, gelatin, hydroxy cellulose, acacia, polyethylene glycol, mannitol, sodium chloride, or sodium citrate.

Alternatively, these compounds may be encapsulated, tableted, or formulated in an emulsion or syrup for oral administration. Pharmaceutically acceptable solid or liquid carriers may be added to enhance or preserve the composition, or to facilitate the preparation of the composition. Solid carriers include starch, lactose, calcium sulfate dihydrate, kaolin, magnesium stearate or stearic acid, talc, pectin, acacia, agar or gelatin. Liquid carriers include syrup, peanut oil, olive oil, saline, and water. The carrier may also include a sustained release material such as glyceryl monostearate or glyceryl distearate, alone or with a wax. The amount of solid carrier varies but will preferably be between about 20 mg to about 1 g per dosage unit. The pharmaceutical preparations are prepared according to conventional pharmaceutical techniques including milling, mixing, granulating and compressing for tablets when necessary; or milling, mixing and filling for hard gelatin capsules. When a liquid carrier is used, the preparation will take the form of a syrup, elixir, emulsion, or an aqueous or non-aqueous suspension. Such a liquid preparation may be administered orally directly or after filling into a soft gelatin capsule.

For rectal administration, the compounds of the present invention can also be combined with excipients such as cocoa butters, glycerin, gelatin or polyethylene glycols and made into a suppository.

For topical administration, the compounds of the present invention can be combined with diluents to form ointments, gels, pastes, creams, powders or sprays. Compositions which are ointments, gels, pastes or creams contain diluents, for example animal and vegetable fats, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures of these substances. Compositions which are powders or sprays contain diluents, for example, lactose, talc, silicic acid, aluminum hydroxide, calcium silicate and polyamide powder, or mixtures of these substances. Additionally, for topical ocular administration, typical carriers are water, mixtures of water and water-miscible solvents, such as lower alkanols or vegetable oils, and water-soluble non-toxic polymers, for example cellulose derivatives such as methyl cellulose.

The compounds described herein are Fab I inhibitors and are useful for the treatment of bacterial infections. For example, these compounds are useful in the treatment of bacterial infections, such as, for example

Upper respiratory tract infections (e.g. otitis media, bacterial tracheitis, acute epiglottitis, thyroiditis), lower respiratory tract infections (e.g. empyema, lung abscess), heart (e.g. endocarditis caused by infectious agents) , gastrointestinal (e.g. secretory diarrhea, spleen abscess, retroperitoneal abscess), CNS (e.g. brain abscess), eyes (e.g. blepharitis, conjunctivitis, keratitis, endophthalmitis, pre-septal and orbital cellulitis, tear apparatus inflammation), kidneys and ureters (e.g. epididymitis, intrarenal and perirenal abscess, toxic shock syndrome), skin (e.g. impetigo, lymphadenitis of the hair follicles, skin abscesses, cellulitis, wound infection, bacterial myositis) and bones and joints (e.g. sepsis arthritis, osteomyelitis).

The compounds of the present invention may also be useful as antifungal agents. Additionally, the compounds may be useful in combination with known antibiotics.

The compounds of the present invention are administered to the patient such that the drug concentration is effective for treating bacterial infections. The pharmaceutical composition containing the compound is administered in an oral dose of from about 10 mg to about 1000 mg, taken once or several times a day, in a manner compatible with the patient's condition.

Preferably, the oral dose may be about 50 mg to about 500 mg, although the dosage may be varied depending upon the age, body weight, and symptoms of the patient. Parenteral administration is preferred for the treatment of acute infections.

The most effective is the intravenous infusion of the compound of formula (I) in 5% dextrose in water or isotonic saline, or a similar formulation with suitable excipients, although bolus intramuscular injection is also useful. One skilled in the art will readily determine the exact level and mode of administration of the compounds.

The compounds can be tested in one of several bioassays to determine the concentration of the compound that is required to obtain a given pharmacological effect.

Cloning of Fabl from S. aureus

The phabl gene was cloned from the chromosomal DNA of S. aureus strain WCUH29 using the polymerase chain reaction. Amplification was performed using DNA Tag polymerase (BRL) and the following primers:

5'-CGCCTCGAGATGTTAAATCTTGAAAACAAAACATATGTC-3 'and

5'-CGCGGATCCAATCAAGTCAGGTTGAAATATCCA-3 '(Xhol and BamHI sites of action are underlined). The obtained fragment was then digested with

Xhol and BamHI and ligated into Xhol and BamHI digested pET-16b expression vector (Novagen) to generate pET-His10-fabI. The sequence of the fabl genes was confirmed by automated cycle sequencing using an Applied Biosystems model 377 machine. A tagless version of pET-fabl was constructed by digesting pET-His10-fabI with NcoI and NdeI to remove the 97 bp fragment encoding the His 10 tag, the factor Xa cleavage site and the first 8 amino acids of Fabl, and replace them with a linker encoding the first 8 amino acids of FabI plus the remainder. glycine between the initiator methionine and lysine at position 2. This plasmid was named pET-phabl. The linker was made by combining the following two oligonucleotides:

5'-CATGGGCTTAAATCTTGAAAACAAAACA-3 'and

5'-TATGTTTTGTTTTCAAGATTTAAGCC-3 '

The linker sequence in pET-fabI was confirmed by didesoxy sequencing. Only native FabI was used for compound evaluation. For the overproduction of native FabI, the Petfabl plasmid was transformed into BL21 (DE3) cells (Novagen) to give rise to strain BL21 (DE3): pET-phabl.

Purification of FabI from S. aureus

S. aureus FabI was expressed as a soluble protein up to 10% of the total cell protein content, yielding 400 g of cells from 15 L fermentation in tryptone-phosphate medium. Cells were lysed and a sample was centrifuged. The resulting supernatant was filtered and purified using three consecutive chromatography columns: an ion exchange chromatography column (Sourse 15Q), dye affinity (Blue Sepharose) and size exclusion (Superose 12). After each column, the FabI-containing fractions were pooled, concentrated, and checked for purity and biological activity.

PL 201 627 B1

FabI cloning from E. coli

A PCR fragment of the size specific to E. coli FabI was PCR amplified from E. coli chromosomal DNA, cloned into the TOPO TA cloning vector and checked by colony PCR + restriction endonuclease analysis. The putative PCR fragment for E. coli FabI was cloned into the pBluePet expression vector. The FabI clone was transformed into E. coli BL21 (DE3) strain.

Small scale expression studies showed a band of overexpressed protein with a molecular weight (~ 28 KDa) specific to E. coli FabI clearly visible after Coomassie staining of SDS PAGE gels. DNA sequencing of the E. coli FabI expression constructs showed that no errors were seen. N'-terminal amino acid sequencing confirmed that the overexpressed protein band was E. coli FabI.

Purification of FabI from E. coli

E. coli FabI was expressed as a soluble protein up to 15% of the total cell protein content, yielding 120 g of cells from 3 L fermentation flasks in modified Terrific Broth. Cells were lysed and a sample was centrifuged. The resulting supernatant was filtered and purified using three successive chromatography columns: ion exchange (Sourse 15Q), dye affinity (Blue Sepharose) and size exclusion (Superose 12). After each column, the FabI-containing fractions were pooled, concentrated, and checked for purity and biological activity.

S. aureus FabI enzyme inhibition assay (NADH)

The determinations were made on half the surface of 96-well microtiter plates. Compounds were evaluated in 50 µL of the test mixture containing 100 mM NaADA, pH 6.5 (ADA = N- [2-acetamido] -2-iminodiacetic acid), 4% glycerol, 0.25 mM crotonoyl CoA, 1 mM NADH and the appropriate FabI dilution from S. aureus. Inhibitor concentrations were typically varied in the range of 0.01-10 µM. The consumption of NADH was monitored for 20 minutes at 30 ° C by following the change in absorbance at 340 nm. Initial rates were estimated from the exponential fit of nonlinear progress curves as the slope of the tangent at t = 0 min. IC50 values ​​were estimated from fitting the initial rates to a standard four parameter model and are typically reported as mean ± standard deviation in duplicate determinations. Triclosan, a commercial antimicrobial agent and FabI inhibitor, is now included in all assays as a positive control. The compounds of the present invention have IC50 values ​​from about 5.0 µM to about 0.05 µM.

S. aureus FabI enzyme inhibition assay (NADPH)

The determinations were made on half the surface of 96-well microtiter plates. Compounds were evaluated in 150 µL of the test mixture containing 100 mM NaADA, pH 6.5 (ADA = N- [2-acetamido] -2-iminodiacetic acid), 4% glycerol, 0.25 mM crotonoyl CoA, 50 µM NADPH and the appropriate FabI dilution from S. aureus. Inhibitor concentrations were typically varied in the range of 0.01-10 µM. The consumption of NADPH was monitored for 20 minutes at 30 ° C by following the change in absorbance at 340 nm. Initial rates were estimated from the exponential fit of nonlinear progress curves as the slope of the tangent at t = 0 min. IC50 values ​​were estimated from fitting the initial rates to a standard four parameter model and are typically reported as mean ± standard deviation of duplicate determinations. Triclosan, a commercial antimicrobial agent and FabI inhibitor, is now included in all assays as a positive control.

E. coli FabI enzyme inhibition assay

The determinations were made on half the surface of 96-well microtiter plates. Compounds were evaluated in 150 µL of the test mixture containing 100 mM NaADA, pH 6.5 (ADA = N- [2-acetamido] -2-iminodiacetic acid), 4% glycerol, 0.25 mM crotonoyl CoA, 50 µM NADH and the appropriate dilution of E. coli FabI. Inhibitor concentrations were typically varied in the range of 0.01-10 µM. The consumption of NADH was monitored for 20 minutes at 30 ° C by following the change in absorbance at 340 nm. Initial rates were estimated from the exponential fit of nonlinear progress curves as the slope of the tangent at t = 0 min. IC50 values ​​were estimated from fitting the initial rates to a standard four parameter model and are typically reported as mean ± standard deviation of duplicate determinations. Triclosan, a commercial antimicrobial agent and FabI inhibitor, is now included in all assays as a positive control. Compounds of the present invention have 1059 values ​​from about 100.0 µM to about 0.05 µM.

Preparation and purification of crotonoyl ACP

Reactions contained 5 mg / mL E. coli apo-ACP, 0.8 mM crotonoyl CoA (Fluka), 10 mM MgCl2, and 30 µM S. pneumoniae ACP synthase in 50 mM NaHEPES, pH 7.5. The mixture was gently stirred

On a magnetic stirrer at 23 ° C for 2 h, the reaction was terminated by adding 15 mM EDTA. The reaction mixture was filtered through a 0.2 micron filter and applied to a MonoQ column (Pharmacia) equilibrated with 20 mM Tris-Cl, pH 7.5. The column was washed with buffer until all unbound material was removed (as observed by UV detection) and crotonoyl-ACP was eluted with a linear gradient from 0 to 400 mM NaCl.

Assay for inhibition of S. aureus FabI enzyme using crotonoyl-ACP

The determinations are made on half the surface of 96-well microtiter plates. Compounds are assessed in 150 µL of the test mixture containing 100 mM NaADA, pH 6.5 (ADA = N- (2-acetamido) -2-iminodiacetic acid), 4% glycerol, 25 µM crotonoyl-ACP, 50 µM NADPH and an appropriate dilution Fab I from S. aureus (approximately 20 nM). Inhibitor concentrations typically vary between 0.01-10 µM. The consumption of NADPH is monitored for 20 minutes at 30 ° C by following the change in absorbance at 340 nm. Initial speeds are estimated from the linear fit of the progress curves. IC50 values ​​are estimated from fitting the initial rates to a standard four parameter model (Equation 1) and are typically reported as mean ± standard deviation of duplicate determinations. Compounds of the present invention have IC50 values ​​from about 100.0 µM to about 0.04 µM in this assay. The apparent Ki is calculated from Equation 2 assuming that the inhibition is in competition with crotonoyl-ACP.

Equation 1: v = Range / (1 + [I] / IC50) s + Background

Equation 2: Ki (pos.) = IC50 / (1 + [S] / Ks)

FabK enzyme inhibition assay

FabK catalyzes the reduction of enoyl-ACPs with the simultaneous oxidation of NADH. The reduction of crotonoyl-ACP to butyryl-ACP can be monitored by observing the change in absorbance at 340 nm as the oxidation of NADH.

Assays were performed on Half area Costar 3696 plates in a final assay volume of 150 µL on a Spectramax plate reader. Substrates (NADH and crotonoyl-ACP) were incubated with Fab K enzyme in 100 mM N- [2-acetamido] -2-iminodiacetic acid (ADA), pH 6.5, 100 mM NHCl, 4% glycerol at 30 ° C and the reaction was monitored at 340 nm.

Using the above assay, inhibition of Fab K by the compounds was tested. 30 µL of inhibitor was added to a well of the plate. Then 30 µL of 250 µM NADH stock and 60 µL of 67.5 µM crotonoyl ACP stock were added to the well. The plate was incubated at 30 ° C for 5 min. The reaction was initiated by adding 30 µL of 6.25 nM enzyme stock to the well (also pre-incubated at 30 ° C). The reaction was then monitored at A340 nm for 30 min at 30 ° C. Positive controls were reactions with no relationship. Negative controls were enzyme-free and compound-free reactions. The final concentrations in the assay mixture were 25 µM crotonoyl ACP, 50 µM NADH, and 1.25 nM enzyme.

IC50 values ​​for compounds were determined by performing the assay at 8 different compound concentrations (100 µM-0.75 µM) in duplicate. IC50 was calculated using Grafit software (version 4.09). Two Fab K inhibitors of the present invention have IC50 values ​​of about 5 µM.

Determination of microbicidal activity

Whole cell microbicidal activity was determined by broth microdilution using the procedure recommended by the National Committee for Clinical Laboratory Standards (NCCLS), Document M7-A4, "Methods for Dilution Susceptibility Tests for Bacteria that Grow Aerobically." The compound was tested in serial two-fold dilutions ranging from 0.06 to 64 µg / mL. The test organisms were selected from the following laboratory strains: Staphylococcus aureus Oxford, Staphylococcus aureus WCUH29, Streptococcus pneumoniae ERY2, Streptococcus pneumoniae 1629, Streptococcus pneumoniae N 1387, Enterococcus faecalls I, Enterococcus aureus faecalis l, Haemophilus influencer2, Haemophilus1, Haemophilus influ. coli 7623 AcrABEFD +, Escherichia coli 120 AcrAB-, Escherichia coli MG1655, Escherichia coli MG1658. The minimum inhibitory concentration (MIC) was defined as the lowest concentration of compound that inhibited visible growth. A mirror reader was used to assist in determining the MIC endpoint.

One skilled in the art would consider any compound with an MIC less than 256 µg / mL as a potential candidate compound. Preferably, the compounds used in the microbicidal assays of the present invention

Of the present invention have a MIC value of less than 128 µg / mL. Most preferably, said compounds have a MIC of less than 64 µg / mL.

In accordance with the present invention, preferred Fab I and Fab K enzyme inhibition assays use crotonoyl-ACP as substrate instead of crotonoyl-CoA. Thus, the present invention covers the production and purification of crotonoyl-ACP and the use of this purified enzyme in Fab I and Fab K inhibition assays. Crotonoyl-ACP was synthesized using ACP synthase from S. the apo-acyl carrier (ACP) from E. coli. In a further aspect of the present invention, it is contemplated that an apo-acyl carrier protein from any bacterial species such as Escherichia coli, Staphylococcus and Streptococcus may be used to produce crotonoyl ACP. This synthesis was performed in the presence of magnesium chloride in NaHEPES, pH 7.5. The reaction is completed in 2 hours at a reaction temperature of about 20-30 ° C, preferably 23 ° C.

The purified crotonoyl-ACP prepared above is then used in the Fab I and Fab K assays to determine inhibitors of the present invention. For example, assays can be performed on Half-surface Costar 3696 plates, preferably with a final assay volume of 150 [mu] l on a Spectramax plate reader. Preferred substrates for use in the methods of the invention are NADH, NADPH, an NADH analog, and crotonoyl ACP.

The invention further relates to preferred methods comprising the step of incubating the substrates with Fab I or Fab K in 100 mM N- [2-acetamido] -2 iminodiacetic acid (ADA), pH 6.5. This reaction can be controlled at 340 nm, including wavelengths.

The following examples are not intended to limit the scope of the present invention in any way, but are intended to illustrate how to make and use the compounds of the present invention. Many other embodiments will be readily apparent to those skilled in the art.

Examples

General information

Proton Magnetic Resonance Spectra (¹H NMR) were recorded at 300 or 360 MHz and chemical shifts are reported in parts per million (δ) with field strengths below tetramethylsilane (TMS) as internal standard.

Abbreviations for NMR data mean as follows: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, dd = doublet of doublets, dt = doublet of triplets, pos. = apparent, br = broad. J is the NMR coupling constant measured in Hertz.

CDCl3 is deuteriochloroform, DMSO-d6 is hexadeuteriodimethylsulfoxide, and CD3OD is tetradeuteriomethanol. Mass spectra were obtained using electrospray (ES) ionization techniques. Elemental analyzes were performed by Quantitative Technologies Inc., Whitehouse, NJ. Melting points were obtained with a Thomas-Hoover melting point apparatus and are uncorrected. All temperatures are given in degrees Celsius. Analtech Silica Gel GF and E. Merck Silica Gel 60 F-254 thin layer plates were used for thin layer chromatography. Flash chromatography was performed on E. Merck Kieselgel 60 silica gel (230-400 mesh). Analytical HPLC was performed on Beckman chromatography systems. Preparative KPLC was performed using Gilson chromatography systems. ODS refers to an octadecylsilyl derivative of a silica gel derivative as chromatographic support. YMC ODS-AQ^® indicates an ODS chromatographic support and is a registered trademark of YMC Co. Ltd., Kyoto, Japan. PRP-1^® designates a polymeric (styrene-divinylbenzene) chromatographic support and is a registered trademark of Hamilton Co., Reno, Nevada. Celite^® is a filter aid composed of acid washed diatomaceous silica and is a registered trademark of Manville Corp., Denver, Colorado.

Production 1

Preparation of 1-methyl-2- (methylaminomethyl) -1H-indole

a) Ethyl 1-methyl-1H-indole-2-carboxylate

NaH (60% dispersion in mineral oil, 8.02 g, 200.49 mmol) was washed with hexane then suspended in dry DMF (530 mL). Solid ethyl indole-2-carboxylate (25.29 g, 133.66 mmol) was added portionwise over 5-10 min, allowing gas evolution to subside between additions. When the addition was complete, the yellow mixture was stirred for 15 min, then all methyl iodide (42 mL, 668.3 mmol) was added all at once. The reaction was exothermic and the internal temperature rose to 40-45 ° C. After 1 h, the reaction was quenched with 10% NH4Cl (100 mL) and concentrated

PL 201 627 B1 on a rotary evaporator (high vacuum). The residue was partitioned between Et2O (500 mL) and H2O (100 mL) and the layers were separated. The Et2O layer was washed with H2O (100 mL), dried (MgSO4) and concentrated to give the title compound (27.10 g, quantitative) as a light yellow solid. It was used without further purification.

TLC (10% EtOAc / hexane) R f = 0.39.

b) N, 1-Dimethyl-1H-indole-2-carboxamide

A suspension of ethyl 1H-indole-2-carboxylate (27.10 g, 133.34 mmol) in 40% aqueous CH3NH2 (300 mL) and Moss (30 mL) was stirred at room temperature. The solid tended to gradually creep up the sides of the flask, and was rinsed periodically with MeOH. The flask was tightly capped to retain the material inside the flask. As the reaction proceeded, the solid dissolved but eventually the product began to precipitate. The reaction was stirred at room temperature for 5 days then concentrated to remove approximately 200 mL of the solvent. The residue was diluted with H2O (300 mL) and the solid was collected by filtration and washed with H2O. Drying at 50-60 ° C under high vacuum gave the title compound (23.45 g, 93%) as a pale yellow solid.

¹H NMR (300 MHz, CDCl3) δ 7.63 (d, J = 8.0 Hz, 1H), 7.27-7.43 (m, 2H), 7.10-7.20 (m, 1H) , 6.80 (s, 1H), 6.10-6.30 (m, 1H), 4.06 (s, 3H), 3.01 (d, J = 4.9 Hz, 3H).

c) 1-Methyl-2- (methylaminomethyl) -1H-indole

A 3-liter 3-neck round bottom flask equipped with an overhead stirrer was charged with N, 1-dimethyl-1H-indole-2-carboxamide (23.45 g, 124.58 mmol) and anhydrous THF (170 mL). The solution was stirred while a solution of LiAlH4 in THF (1.0 M, 250 mL, 250 mmol) was added via a syringe. Gas was evolved during the addition of the first 50 mL of LiAlH4 solution. When the addition was complete, the resulting pale yellow solution was heated to gentle reflux. After 23 h, the reaction was cooled in ice and quenched by the sequential dropwise addition of H 2 O (9.5 mL), 15% NaOH, 9.5 mL) and H 2 O (28.5 mL). The mixture was stirred for 15 min, then filtered through Celite^® and the filter pad was washed thoroughly with THF. The filtrate was concentrated and the residue was flash chromatographed on silica gel (10% MeOH / CHCl3 containing 0.5% conc. NH4OH). The title compound (20.17 g, 93%) was obtained as a light yellow oil.

¹H NMR (300 MHz, CDCl3) δ 7.56 (d, J = 7.8 Hz, 1H), 7.02-7.35 (m, 3H), 6.38 (s, 1H), 3.88 (s, 2H), 3.75 (s, 3H), 2.49 (s, 3H).

Production 2

Preparation of (E) -3- (6-aminopyridin-3-yl) acrylic acid (Method A)

a) Benzyl (E) -3- (6-aminopyridin-3-yl) acrylate

A solution of 2-amino-5-bromopyridine (2.25 g, 13.0 mmol), benzyl acrylate (3.2 g, 19.7 mmol), Pd (OAc) 2 (0.31 g, 1.4 mmol) , tri-ortho-tolylphosphine (0.73 g, 2.4 mmol) and diisopropylethylamine (3.5 mL, 20.0 mmol) in propionitrile (50 mL) was heated at reflux overnight. The dark mixture was filtered through Celite^® and the filtrate was concentrated. Flash chromatography on silica gel (3% MeOH / CH2Cl2) gave the title compound (1.3 g, 39%).

MS (ES) m / e 255 (M + H)⁺.

b) (E) -3- (6-Aminopyridin-3-yl) acrylic acid

A solution of benzyl (E) -3- (6-aminopyridin-3-yl) acrylate (1.3 g, 5.1 mmol) and 1.0 N NaOH (10 mL, 10 mmol) in MeOH was refluxed overnight . The solution was concentrated under reduced pressure and the residue was dissolved in H2O. The pH was adjusted to 6 with dilute HCl and the solid precipitate was collected by filtration and dried to afford the title compound (0.6 g, 72%) as a white solid.

MS (ES) m / e 165 (M + H)⁺.

Production 3

Preparation of (E) -3- (6-aminopyridin-3-yl) acrylic acid (Method B)

a) (E) -3- (6-Aminopyridin-3-yl) acrylic acid

Acrylic acid (23 mL, 0.33 mol) was carefully added to a solution of 2-amino-5-bromopyridine (25.92 g, 0.15 mol) and Na2CO3 (55.64 g, 0.53 mol) in H2O (600 mL). Then PdCl2 (0.53 g, 0.003 mol) was added and the mixture was heated to reflux. After 24 h, the reaction was cooled to room temperature and filtered, and the filtrate was adjusted to pH 6 with aqueous HCl. Additional H 2 O (0.5 L) was added to improve mixing and the mixture was stirred for 1 h. The pH was re-adjusted to 6, and the solid was collected by vacuum filtration. The filter pad was washed sequentially with H 2 O (2 x 0.5 L), cold absolute EtOH

PL 201 627 B1 (100 mL) and Et2O (2 x 250 mL). Drying under high vacuum at elevated temperature gave the title compound (15.38 g, 62%) as a beige solid.

¹H NMR (300 MHz, DMSO-d6) δ 8.11 (d, J = 2.0 Hz, 1H), 7.75 (dd, J = 8.7, 2.0 Hz, 1H), 7.43 (d, J = 15.8 Hz, 1H), 6.53 (s, 2H), 6.45 (d, J = 8.7 Hz, 1H), 6.22 (d, J = 15.8 Hz , 1H).

MS (ES) m / e 165 (M + H)⁺.

Production 4

Preparation of 1-methyl-3- (methylaminomethyl) -1H-indazole

a) Methyl (1-Methyl-1H-indazole) carboxylate

Indazole-3-carboxylic acid (5.0 g, 30 mmol), K2CO3 (12.4 g, 90 mmol) and MeI (9.3 mL, 150 mmol) were combined in dry DMF (100 mL) and heated to 50 ° C C. After 18 h, the mixture was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in EtOAc and filtered, and the filtrate was concentrated under reduced pressure. The residue was chromatographed on silica gel (25% EtOAc / hexane) to provide the title compound (3.88 g, 68%) as a yellow solid.

¹H NMR (300 MHz, CDCl3) δ 8.24 (m, 1H), 7.47 (m, 2H), 7.34 (m, 1H), 4.19 (s, 3H), 4.05 (s , 3H).

b) N, 1-Dimethyl-1H-indazole-3-carboxamide

A suspension of methyl (1-methyl-1H-indazole) carboxylate (3.88 g, 20.4 mmol) in 40% aqueous CH3NH2 (100 mL) and MeOH (5 mL) was stirred at room temperature for 4 h. the suspension became a solution. The mixture was concentrated to approximately 1/3 volume when the product crashed out as a pale yellow solid. The solid was collected by filtration, washed with H 2 O, and dried under reduced pressure to obtain the title compound (3.42 g, 89%) which was pure enough for use in the next step.

¹H NMR (300 MHz, CDCl3) δ 8.24 (m, 1H), 7.47 (m, 2H), 7.34 (m, 1H), 6.95 (bs, 1H), 4.19 (s , 3H), 3.05 (d, J = 12.0 Hz, 3H).

c) 1-Methyl-3- (methylaminomethyl) -1H-indazole

To a solution of N, 1-dimethyl-1H-indazole-3-carboxamide (3.42 g, 18 mmol) in dry THF (90 mL) was added slowly a solution of LiAlH4 in THF (1M, 36 mL, 36 mmol) at room temperature. After 2 h, the mixture was heated to a gentle reflux. After 4 h, the mixture was cooled to room temperature and quenched by dropwise addition of 2M NaOH until a white solid formed. The mixture was dried (MgSO4), filtered and concentrated under reduced pressure to provide the title compound (3.28 g, 100%) as an oil which was pure enough for use in the next step.

MS (ES) m / e 176 (M + H)⁺.

Production 5

Preparation of (E) -3- (3,4-dihydro-2H-pyrido [3,2-b] -1,4-oxazin-7-yl) acrylic acid

a) 3,4-Dihydro-2H-pyrido [3,2-b] -1,4-oxazine

To a suspension of 2H-pyrido [3,2-b] -1,4-oxazin-3 (4H) -one (2.0 g, 13.3 mmol) in dry THF (40 mL) was slowly added a solution of LiAlH4 in THF ( 1.0 M, 26.6 mL, 26.6 mmol) at 0 ° C. After 1 h, the reaction was quenched with 2.0 M NaOH until solid formed. The mixture was dried (MgSO4), filtered and concentrated under reduced pressure to provide the title compound (1.44 g, 79%) as a white solid that was pure enough for use in the next step.

MS (ES) m / e 137 (M + H)⁺.

b) 4- (tert-Butoxycarbonyl) -3,4-dihydro-2H-pyrido [3,2-b] -1,4-oxazine

For a solution of 3,4-dihydro-2H-pyrido [3,2-b] -1,4-oxazine (1.44 g, 10.6 mmol) and di-tert-butyl dicarbonate (2.78 g, 12.7 mmol) ) in dry THF (50 mL), a solution of LiHMDS in THF (1.0 M, 12.7 mL, 12.7 mmol) was added dropwise at 0 ° C. After 30 min, the reaction was quenched with saturated NH4Cl and extracted with EtOAc (3 x). The combined organic layers were dried (MgSO4), filtered and concentrated. Flash chromatography on silica gel (40% EtOAc / hexane) gave the title compound (2.0 g, 80%) as clear oil.

MS (ES) m / e 237 (M + H)⁺.

c) 4- (tert-Butoxycarbonyl) -7-bromo-3,4-dihydro-2H-pyrido [3,2-b] -1,4-oxazine

To a solution of 4- (tert-butoxycarbonyl) -3,4-dihydro-2H-pyrido [3,2-b] -1,4-oxazine (2.0 g, 8.46 mmol) in MeOH (40 mL) was added droplets of Br2 (0.53 mL, 10.2 mmol) at 0 ° C. After 1 h, the mixture was concentrated. The residue was dissolved in 1: 1 Et2O / hexane and filtered. The filtrate was concentrated under reduced pressure to provide the title compound (1.27 g, 48%) as an oil which solidified under reduced pressure.

¹H NMR (400 MHz, CDCl3) δ 8.10 (s, 1H), 7.33 (s, 1H), 4.25 (m, 2H), 3.92 (m, 2H), 1.54 (s , 9H).

PL 201 627 B1

d) (E) -3- [4- (tert-butoxycarbonyl) -3,4-dihydro-2H-pyrido [3,2-b] -1,4-oxazin-7-yl] acrylic acid

A solution of 4- (tert-butoxycarbonyl) -7-bromo-3,4-dihydro-2H-pyrido [3,2-b] -1,4-oxazine (1.27 g,

4.03 mmol), benzyl acrylate (785 mg, 4.84 mmol), Pd (OAc) 2 (45 mg, 0.20 mmol), P (o-tolyl) 3 (122 mg, 0.4 mmol) and (i-Pr) 2NEt (1.76 mL, 10.1 mmol) in propionitrile (20 mL) was degassed (3 x N2 / vacuum) and then heated to reflux. After 18 h, the mixture was cooled to room temperature and concentrated. Flash chromatography on silica gel (25% EtOAc / hexane) gave the title compound (1.17 g, 73%) as a yellow oil.

MS (ES) m / e 397 (M + H)⁺.

e) (E) -3- (3,4-Dihydro-2H-pyrido [3,2-b] [1,4] oxazin-7-yl) acrylic acid

(E) -3- [4- (tert-butoxycarbonyl) -3,4-dihydro-2H-pyrido [3,2-b] -1,4-oxazin-7-yl] acrylic acid (1.17 g, 2.95 mmol) was dissolved in 4N HCl in dioxane (15 mL). After 72 h, the mixture was concentrated. The residue was dissolved in 1: 1 MeOH / H2O (20 mL). 1.0 N LiOH (15 mL, 15 mmol) was added and the mixture was heated to reflux. After 18 h, the mixture was cooled to room temperature and concentrated to approximately 1/3 volume. The mixture was adjusted to pH 6 with 10% HCl. The solid was collected by filtration, washed with H 2 O, and dried under reduced pressure to obtain the title compound (315 mg, 52% over 2 steps).

MS (ES) m / e 207 (M + H)⁺.

Production 6

Preparation of (E) -3- (5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl) acrylic acid

a) 1,2,3,4-Tetrahydro-1,8-naphthyridine

1,8-Naphthyridine (1.0 g, 7.68 mmol) was hydrogenated (50 psi) with 10% Pd / C (100 mg) in absolute ethanol (40 mL) for 18 h. The mixture was filtered through a pad of Celite^® and the filtrate was concentrated to give the title compound (1.04 g) which was pure enough for use in the next step.

MS (ES) m / e 135 (M + H)⁺.

b) 1- (tert-Butoxycarbonyl) -1,2,3,4-tetrahydro-1,8-naphthyridine

To a solution of 1,2,3,4-tetrahydro-1,8-naphthyridine (1.04 g, 7.68 mmol) and di-tert-butyl dicarbonate (2.01 g, 9.22 mmol) in dry THF ( 40 mL) was added a solution of LiHMDS in THF (1.0 M, 9.22 mL, 9.22 mmol) dropwise at 0 ° C. After 30 min, the reaction was quenched with saturated NH4Cl and extracted with EtOAc (3 x). The combined organic layers were dried (MgSO4), filtered and concentrated. Flash chromatography on silica gel (40% EtOAc / hexane) gave the title compound (1.37 g, 76% over 2 steps) as an orange oil which solidified under reduced pressure.

¹H NMR (400 MHz, CDCl3) δ 8.33 (m, 1H), 7.37 (m, 1H), 6.94 (m, 1H), 3.77 (m, 2H), 2.75 (t , J = 6.5 Hz, 2H), 1.93 (m, 2H), 1.54 (s, 9H).

c) 1- (tert-Butoxycarbonyl) -6-bromo-1,2,3,4-tetrahydro-1,8-naphthyridine

Glacial HOAc (3.4 mL) was added to a solution of 1- (tert-butoxycarbonyl) -1,2,3,4-tetrahydro-1,8-naphthyridine (1.37 g, 5.85 mmol) in CH2Cl2 (30 mL). , 58.5 mmol) and NBS (1.09 g, 6.14 mmol). After 72 h, the mixture was washed with 2.0 M NaOH, H 2 O, and brine. The mixture was dried (MgSO4), filtered and concentrated under reduced pressure to provide the title compound (1.79 g, 98%) which was pure enough for use in the next step.

¹H NMR (400 MHz, CDCl 3) δ 8.35 (s, 1H), 7.51 (s, 1H), 3.77 (m, 2H), 2.75 (t, J = 6.5 Hz, 2H ), 1.93 (m, 2H), 1.54 (s, 9H).

d) Benzyl (E) -3- [8- (tert-Butoxycarbonyl) -5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl] acrylate

A solution of 1- (tert-butoxycarbonyl) -6-bromo-1,2,3,4-tetrahydro-1,8-naphthyridine (1.79 g, 5.70 mmol), benzyl acrylate (1.11 g, 6, 34 mmol), Pd (OAc) 2 (65 mg, 0.29 mmol; P (o-tolyl) 3 (173 mg, 0.57 mmol) and (i-Pr) 2NEt (2.5 mL, 14.25 mmol) in propionitrile (30 mL) was degassed (3 x N2 / vacuum) then heated to reflux After 18 h the mixture was cooled to room temperature and concentrated. Flash chromatography on silica gel (25% EtOAc / hexane) gave the title compound (1.21 g, 54%) as a yellow solid.

¹H NMR (400 MHz, CDCl 3) δ 8.44 (s, 1H), 7.65 (d, J = 16.0 Hz, 1H), 7.53 (s, 1H), 7.40 (m, 5H ), 6.43 (d, J = 16.0 Hz, 1H), 5.25 (s, 2H), 3.77 (m, 2H), 2.75 (t, J = 6.5 Hz, 2H ), 1.93 (m, 2H), 1.54 (s, 9H).

e) (E) -3- (5,6,7,8-Tetrahydro-1,8-naphthyridin-3-yl) acrylic acid (E) -3- [8- (tert-Butoxycarbonyl) -5.6, Benzyl 7,8-tetrahydro-1,8-naphthyridin-3-yl] acrylate (1.21 g, 3.07 mmol) was dissolved in 4N HCl in dioxane (15 mL). After 18 h, the mixture was concentrated. The residue was dissolved in 1: 1 MeOH / H2O (15 mL). 1.0 N LiOH (15 mL, 15 mmol) was added and the mixture was heated to reflux. After 18 h, the mixture was cooled to room temperature and concentrated

PL 201 627 B1 to approximately 1/3 volume. The mixture was adjusted to pH 6 with 10% HCl. The solid was collected by filtration, washed with H 2 O, and dried under reduced pressure to obtain the title compound (180 mg, 29% over 2 steps).

MS (ES) m / e 205 (M + H)⁺.

Production 7

Preparation of 2- (methylaminomethyl) thieno [2,3-b] thiophene

a) 3- (1,3-Dioxolan-2-yl) thiophene

Ethylene glycol (25 mL, 445.8 mmol) and p-toluenesulfonic acid hydrate (848 mg, 4.458 mmol) were added to a solution of thiophene-3-carboxaldehyde (5.0 g, 44.58 mmol) in benzene (200 mL). The mixture was heated to reflux under a Dean-Stark separator. After 18 h the mixture was cooled to room temperature, washed with saturated NaHCO 3 then H 2 O, dried (MgSO 4) and concentrated in vacuo to give the title compound (6.32 g, 91%) as a light amber oil.

¹H NMR (400 MHz, CDCl3) δ 7.42 (s, 1H), 7.32 (m, 1H), 7.16 (m, 1H), 5.91 (s, 1H), 4.12-3 .99 (m, 4H).

b) 2- (Carboethoxymethylthio) -3- (1,3-dioxolan-2-yl) thiophene

To a solution of 3- (1,3-dioxolan-2-yl) thiophene (6.32 g, 40.46 mmol) in dry THF (200 mL) was added slowly a solution of n-BuLi in hexane (1.7 M, 28 8 mL, 49 mmol) at -78 ° C. After 30 min, all the sulfur (1.57 g, 49 mmol) was added immediately. After 30 min, ethyl bromoacetate (7.4 mL, 66.87 mmol) was added slowly and after another 30 min the mixture was warmed to room temperature. After 2 h at room temperature, the mixture was concentrated under reduced pressure. The residue was dissolved in Et2O, washed with H2O (3 x), dried (MgSO4) and concentrated to provide the title compound as an oil which was pure enough for use in the next step.

c) 2- (Carboethoxymethylthio) -3-formylthiophene

To a solution of 2- (carboethoxymethylthio) -3- (1,3-dioxolan-2-yl) thiophene (from step b) in acetone (200 mL) was added p-toluenesulfonic acid (761 mg, 4.0 mmol) at room temperature . After 18 h, the mixture was concentrated. The residue was dissolved in Et2O, washed with saturated NaHCO3, H2O (2 x), dried (MgSO4) and concentrated under reduced pressure to provide the title compound as an oil that was pure enough for use in the next step.

d) Ethyl thieno [2,3-b] thiophene-2-carboxylate

DBU (0.6 mL, 4.0 mmol) was added to a solution of 2- (carboethoxymethylthio) -3-formylthiophene (from step c) in MeOH (200 mL) at 0 ° C. After 1 h, the mixture was warmed to room temperature and concentrated. The residue was dissolved in EtOAc, washed with 10% HCl, H 2 O (3 x), dried (MgSO 4) and concentrated. Flash chromatography on silica gel (50% toluene / hexane) gave the title compound (3.84 g, 45% over 4 steps) as an off-white solid¹H NMR (400 MHz, CDCl 3) δ 7.95 (s, 1H), 7.40 (d, J = 5.2 Hz, 1H), 7.26 (d, J = 5.2 Hz, 1H), methyl ester 3.92 (s, 3H), ethyl ester 4.38 (q, J = 7.1 Hz, 2H)) and 1.41 (t, J = 2.4 Hz, 3H).

e) N-Methyl-2- (thieno [2,3-b] thiophene) carboxamide

A suspension of ethyl thieno [2,3-b] thiophene-2-carboxylate (3.84 g, 18.1 mmol) in 40% aqueous CH3NH2 (100 mL) and MeOH (10 mL) was stirred at room temperature for 18 h. During this time, the suspension became a solution.

The mixture was concentrated to approximately 1/3 volume when the product precipitated. The solid was collected by filtration, washed with H 2 O, and dried under reduced pressure to obtain the title compound (3.01 g, 85%).

¹H NMR (400 MHz, d6-DMSO) δ 8.60 (bs, 1H), 7.92 (s, 1H), 7.67 (d, J = 5.2 Hz, 1H), 7.38 (d , J = 5.2 Hz, 1H), 2.78 (d, J = 4.6 Hz, 3H).

f) 2- (Methylaminomethyl) thieno [2,3-b] thiophene

To a solution of N-methyl-2- (thieno [2,3-b] thiophene) carboxamide (3.01 g, 15.26 mmol) in dry THF (75 mL) was slowly added a solution of LiALH4 in THF (1.0 M, 30 mL, 30 mmol) at room temperature. After gas evolution had ceased, the mixture was heated to a gentle reflux. After 18 h the mixture was cooled to room temperature and quenched by dropwise addition of 2.0 M NaOH until a white solid formed.

The mixture was dried over MgSO4, filtered and concentrated under reduced pressure to provide the title compound (2.18 g, 78%) as a brown oil.

¹H NMR (400 MHz, CDCl 3) δ 7.30 (d, J = 5.2 Hz, 1H), 7.15 (d, J = 5.2 Hz, 1H), 7.04 (s, 1H), 4.00 (s, 2H), 2.49 (s, 3H).

PL 201 627 B1

Production 8

Preparation of 2- (methylaminomethyl) thieno [3,2-b] thiophene

a) N-Methyl-2- (thieno [3,2-b] thiophene) carboxamide

EDC (624 mg, 3.26 mmol) was added to a solution of thieno [3.2-b] thiophene-2-carboxylic acid (500 mg, 2.71 mmol), CH3NH2 (2.0 M in THF, 2.7 mL , 5.42 mmol), HOBt.H2O (440 mg, 3.26 mmol), and Et3N (0.95 mL, 6.78 mmol) in dry DMF (14 mL) at room temperature. After 18 h the mixture was diluted with H 2 O and extracted with EtOAc (3 x). The combined organic layers were dried (MgSO4) and concentrated to afford the title compound (415 mg, 78%) which was pure enough for use in the next step.

¹H NMR (400 MHz, CDCl 3) δ 7.70 (s, 1H), 7.52 (d, J = 5.3 Hz, 1H), 7.27 (d, J = 5.3 Hz, 1H), 3.02 (d, J = 4.9 Hz, 3H).

b) 2- (Methylaminomethyl) thieno [3,2-b] thiophene

To a solution of N-methyl-2- (thieno [3,2-b] thiophene) carboxamide (415 mg, 2.1 mmol) in dry THF (10 mL) was slowly added a solution of LiAlH4 in THF (1.0 M, 4. 2 mL, 4.2 mmol) at room temperature. After gas evolution had ceased, the mixture was heated to a gentle reflux. After 18 h the mixture was cooled to room temperature and quenched by dropwise addition of 2.0 M NaOH until a white solid formed. The mixture was dried (MgSO4), filtered and concentrated to provide the title compound (361 mg, 94%) as a brown oil.

¹H NMR (400 MHz, CDCl 3) δ 7.31 (d, J = 5.2 Hz, 1H), 7.21 (d, J = 5.2 Hz, 1H), 7.11 (s, 1H), 4.01 (s, 2H), 2.50 (s, 3H).

Production 9

Preparation of (E) -3- (3H-imidazo [4,5-b] pyridin-6-yl) acrylic acid

a) 5-Bromo-2,3-diaminopyridine

To a suspension of 2-amino-5-bromo-3-nitropyridine (2.0 g, 9.17 mmol) in absolute EtOH (50 mL) was added SnCl2 hydrate (9.3 g, 41.3 mmol) and the mixture was heated. to boiling point. After 3 h, the mixture was cooled to room temperature and concentrated. The residue was dissolved in 2.0 M NaOH and extracted with EtOAc (3 x). The combined organic layers were dried (MgSO4), filtered and concentrated to afford the title compound (1.69 g, 98%) which was pure enough for use in the next step.

MS (ES) m / e 188/190 (M + H)⁺.

b) 6-Bromo-3H-imidazo [4,5-b] pyridine

5-Bromo-2,3-diaminopyridine (1.69 g, 8.99 mmol) was dissolved in 96% formic acid (50 mL) and heated to reflux. After 18 h, the mixture was cooled to room temperature and concentrated. The residue was dissolved in H 2 O and the pH was adjusted to 7 with 2.0 M NaOH. The title compound (1.54 g, 87%) was collected as a solid by filtration, washed with H 2 O and dried under reduced pressure.

MS (ES) m / e 198/200 (M + H)⁺.

c) 6-Bromo-4-trityl-3H-imidazo [4,5-b] pyridine

Et3N (1.3 mL, 9.09 mmol) was added to a suspension of 6-bromo-3H-imidazo [4,5-b] pyridine (1.2 g, 6.06 mmol) in CH2Cl2 (30 mL), followed by trityl chloride (2.03 g, 7.27 mmol) at room temperature. After 72 h the mixture was washed with H 2 O (2 x) and brine then dried (MgSO 4), filtered and concentrated in vacuo to provide the title compound. It was used directly in the next step.

d) Benzyl (E) -3- (4-Trityl-3H-imidazo [4,5-b] pyridin-6-yl) acrylate

A solution of 6-brcmo-4-trityl-3H-imidazo [4,5-b] pyridine (from step a) (6.06 mmol), benzyl acrylate (1.18 g, 7.27 mmol), Pd (OAc) 2 (67 mg, 0.30 mmol), P (o-tolyl) 3 (183 mg, 0.6 mmol), and (i-Pr) 2NEt (2.64 mL, 15.15 mmol) in propionitrile (30 mL) was degassed (3 x N2 / vacuum) then heated to reflux. After 4 h, the mixture was cooled to room temperature and concentrated. Flash chromatography on silica gel (30% EtOAc / hexane) gave the title compound (1.75 g, 55% over 2 steps) as an off white foam.

¹H NMR (400 MHz, CDCl3) δ 8.24 (d, J = 2.0 Hz, 1H), 8.19 (d, J = 2.0 Hz, 1H), 8.06 (s, 1H), 7.77 (d, J = 16.0 Hz, 1H), 7.42-7.11 (m, 20H), 6.48 (d, J = 16.0 Hz, 1H), 5.25 (s , 2H).

d) (E) -3- (3H-imidazo [4,5-b] pyridin-6-yl) acrylic acid (E) -3- (4-Trityl-3H-imidazo [4,5-b] pyridin- Benzyl 6-yl) acrylate (1.75 g, 3.35 mmol) was dissolved in 4N HCl in dioxane (20 mL). After 1 h, the mixture was concentrated. The residue was dissolved in 1: 1 MeOH / H2O (15 mL). 2.0 N NaOH (15 mL, 15 mmol) was added and the mixture was heated to reflux. After 18 h the mixture was cooled to room temperature and concentrated to approximately 1/3

Volume. The mixture was adjusted to pH 4 with 10% HCl. The solid was collected by filtration, washed with H 2 O, and dried under reduced pressure to obtain the title compound (329 mg, 52% over 2 steps) as a white solid.

¹H NMR (400 MHz, d⁶-DMSO) δ 9.10 (s, 1H), 8.94 (s, 1H), 8.84 (s, 1H) and 8.20 (d, J = 16.0 Hz, 1H), 7, 10 (d, J = 16.0 Hz, 1H).

Production 10

Preparation of 6-methyl-5- (methylaminomethyl) -6H-thieno [2,3-b] pyrrole

a) Ethyl (Z) -2-Azido-3- (thiophen-3-yl) acrylate

To a solution of thiophene-3-carboxaldehyde (500 mg, 4.46 mmol) and ethyl 2-azidoacetate (863 mg, 6.69 mmol) in absolute EtOH (20 mL) was added NaOEt (21%, 2.2 mL, 6 69 mmol) at 0 ° C. After 1 h, the reaction was quenched with saturated NH4Cl and extracted with Et2O (3 x).

The combined organic layers were dried (MgSO4), filtered and concentrated. Flash chromatography on silica gel (50% CHCl3 / hexane) gave the title compound (208 mg, 21%) as a pale yellow oil.

¹H NMR (400 MHz, CDCl 3) δ 7.87 (m, 1H), 7.49 (m, 1H), 7.31 (m, 1H), 6.96 (s, 1H), 4.36 (q , J = 7.1 Hz, 2H), 1.39 (t, J = 7.1 Hz, 3H).

b) Ethyl 6H-Thieno [2,3-b] pyrrole-5-carboxylate

A solution of ethyl (Z) -2-azido-3- (thiophen-3-yl) acrylate (208 mg, 0.93 mmol) in xylene (5 mL) was heated to reflux. After 30 min, the mixture was cooled to room temperature and concentrated to afford the title compound (175 mg, 96%) which was pure enough for use in the next step.

¹H NMR (400 MHz, CDCl 3) δ 9.26 (bs, 1H), 7.10 (m, 1H), 7.00 (m, 1H), 6.91 (m, 1H), 4.36 (q , J = 7.1 Hz, 2H), 1.39 (t, J = 7.1 Hz, 3H).

c) N, 6-Dimethyl-6H-thieno [2,3-b] pyrrole-5-carboxamide

To a solution of ethyl 6H-thieno [2,3-b] pyrrole-5-carboxylate (175 mq, 0.9 mmol, see J. Het. Chem. 1984, 21, 215-217) and MeI (0.08 mL , 1.35 mmol) in dry DMF (5 mL) was added NaH (60% dispersion in mineral oil, 43 mg, 1.08 mmol) at 0 ° C. After 2 h, the reaction was quenched with saturated NH4Cl and extracted with EtOAc (3 x). The combined organic layers were dried (MgSO4), filtered, and concentrated to an oil. A solution of the above oil in 40% aqueous CH3NH2 (20 mL) and MeOH (1 mL) was stirred at room temperature for 18 h. The mixture was concentrated to approximately 1/3 volume when the product precipitated. The solid was collected by filtration, washed with H 2 O, and dried under reduced pressure to obtain the title compound (134 mg, 74% over 2 steps).

MS (ES) m / e 195 (M + H)⁺.

d) 6-Methyl-5- (methylaminomethyl) -6H-thieno [2,3-b] pyrrole

To a solution of N, 6-dimethyl-6H-thieno [2,3-b] pyrrole-5-carboxamide (134 mg, 0.69 mmol) in THE dry (5 mL) was added slowly a solution of LiAlH4 in THE (1.0 M , 1.38 mL, 1.38 mmol) at room temperature. After gas evolution had ceased, the mixture was heated to a gentle reflux. After 2 h, the mixture was cooled to room temperature and quenched by dropwise addition of 2M NaOH until a white solid formed. The mixture was dried (MgSO4), filtered and concentrated to afford the title compound as a brown oil (142 mg, 100%) which was pure enough for use in the next step.

¹H NMR (400 MHz, CDCl 3) δ 6.95 (d, J = 5.2 Hz, 1H), 6.78 (d, J = 5.2 Hz, 1H), 6.27 (s, 1H). 3.78 (s, 2H), 3.72 (s, 3H), 2.47 (s, 3H).

Production 11

Preparation of (E) -3- (2-Aminopyrimidin-5-yl) acrylic acid

a) Benzyl (E) -3- (2-Aminopyrimidin-5-yl) acrylate

Follow the procedure for Preparation 2 (a) except that 5-bromo-2-aminopyrimidine (1.95 g, 11.2 mmol) was used in place of 2-amino-5-bromopyridine, prepared the title compound (2.25 g, 79%) as a light orange solid.

MS (ES) m / e 256 (M + H)⁺.

b) (E) -3- (2-Aminopyrimidin-5-yl) acrylic acid

Following the procedure from Preparation 2 (b) except that benzyl (E) -3- (2-aminopyrimidin-5-yl) acrylate (2.93 g, 11.5 mmol) was used instead of (E) -3 Benzyl - (6-aminopyridin-3-yl) acrylate gave the title compound (1.71 g, 90%) as an off-white solid.

MS (ES) m / e 166 (M + H)⁺.

PL 201 627 B1

Production 12

Preparation of (E) -3- (6-aminopyridin-3-yl) -2-methylacrylic acid

a) methyl (E) -3- (6-aminopyridin-3-yl) -2-methylacrylate

Following the procedure in Preparation 2 (a) except that methyl crotonate (4.33 g, 43.3 mmol) was used in place of benzyl acrylate, the title compound (1.0 g, 18%) was prepared as an off-white solid.

MS (ES) m / e 193 (M + H)⁺.

b) (E) -3- (6-Aminopyridin-3-yl) -2-methylacrylic acid

Following the procedure from Preparation 2 (b) except that methyl (E) -3- (6-aminopyridin-3-yl) -2-methylacrylate (1.0 g, 5.2 mmol) was used instead of (E Benzyl) -3- (6-aminopyridin-3-yl) acrylate, the title compound (0.83 g, 90%) was obtained as an off-white solid.

MS (ES) m / e 179 (M + H)⁺.

Production 13

Preparation of (E) -3- (6-amino-2-methylpyridin-3-yl) acrylic acid

a) benzyl (E) -3- (6-Amino-2-methylpyridin-3-yl) acrylate

Following the procedure for Preparation 2 (a) except that 2-amino-5-bromo-6-methylpyridine (5.00 g, 26.7 mmol) was used in place of 2-amino-5-bromopyridine, the title compound was prepared (5.58 g, 78%) as an off-white solid.

MS (ES) m / e 269 (M + H)⁺.

b) (E) -3- (6-Amino-2-methylpyridin-3-yl) acrylic acid

Following the procedure for Preparation 2 (b) except that benzyl (E) -3- (6-amino-2-methylpyridin-3-yl) acrylate (2.20 g, 8.2 mmol) was used instead of ( Benzyl E) -3- (6-aminopyridin-3-yl) acrylate, the title compound (1.31 g, 90%) was prepared as an off-white solid.

MS (ES) m / e 179 (M + H)⁺.

Production 14

Preparation of (E) -3- (6-amino-5-methylpyridin-3-yl) acrylic acid

a) Benzyl (E) -3- (6-Amino-5-methylpyridin-3-yl) acrylate

Following the procedure for Preparation 2 (a) except that 2-amino-5-bromo-3-methylpyridine (5.00 g, 26.7 mmol) was used in place of 2-amino-5-bromopyridine, the title compound was prepared (6.37 g, 89%) as an off-white solid.

MS (ES) m / e 269 (M + H)⁺.

b) (E) -3- (6-Amino-5-methylpyridin-3-yl) acrylic acid

Following the procedure from Preparation 2 (b) except that benzyl (E) -3- (6-amino-5-methylpyridin-3-yl) acrylate (5.00 g, 18.6 mmol) was used instead of ( Benzyl E) -3- (6-aminopyridin-3-yl) acrylate, the title compound (2.98 g, 90%) was prepared as an off-white solid.

MS (ES) m / e 179 (M + H)⁺.

Production 15

Preparation of (E) -3- [6-amino-5- (hydroxymethyl) pyridin-3-yl] acrylic acid

a) 2-Amino-3- (hydroxymethyl) pyridine

Lithium aluminum hydride (300 mL, 1.0 M in THF) was added to a solution of 2-aminonicotinic acid (20.5 g, 148.1 mmol) in THF over 30 minutes. The reaction solution was heated to reflux for 18 h, and then cooled to room temperature. The reaction was quenched by the sequential dropwise addition of H 2 O (11.5 mL), 15% NaOH (11.5 mL), and H 2 O (34.5 mL). The mixture was stirred for 15 min, then filtered through Celite^® and the filter pad was washed thoroughly with THF followed by 5% CH3OH / CHCl3. The filtrate was concentrated to provide the title compound (15.24 g, 83%) as a waxy light yellow solid.

MS (ES) m / e 125 (M + H)⁺.

b) 2-Amino-5-bromo-3- (hydroxymethyl) pyridine

NBS (22.71 g, 127.6 mmol) was added to a solution of 2-amino-3- (hydroxymethyl) pyridine (13.0 g, 116.0 mmol) in CH2Cl2 (300 mL) at room temperature. After stirring at room temperature for 45 min, the reaction solution was concentrated and the residue was dissolved in CHCl3. The resulting suspension was filtered and the filtrate was concentrated to a dark oil. Purification on silica gel (EtOAc) gave the title compound (78%, 18.36 g) as a beige solid.

MS (ES) m / e 204 (M + H)⁺.

PL 201 627 B1

c) Benzyl (E) -3- [6-Amino-5- (hydroxymethyl) pyridin-3-yl] acrylate

Following the procedure for Preparation 2 (a) except that 2-amino-3- (hydroxymethyl) -5-bromopyridine (1.10 g, 5.42 mmol) was used in place of 2-amino-5-bromopyridine, title compound (1.25 g, 81%) as an off-white solid.

MS (ES) m / e 285 (M + H)⁺.

d) (E) -3- [6-Amino-5- (hydroxymethyl) pyridin-3-yl] acrylic acid

Following the procedure for Preparation 2 (b) except that benzyl (E) -3- [6-amino-5- (hydroxymethyl) -pyridin-3-yl] acrylate was used (1.10 g, 5.42 mmol) instead of benzyl t (E) -3- (6-aminopyridin-3-yl) acrylate, the title compound (0.68 g, 65%) was obtained as an off-white solid.

MS (ES) m / e 194 (M + H)⁺.

Production 16

Preparation of 6-bromo-3,4-dihydro-1H-1,8-naphthyridin-2-one

a) 2-Amino-5-bromo-3- (bromomethyl) pyridine hydrobromide

A solution of 2-amino-5-bromo-3-hydroxymethylpyridine (5.00 g, 24.6 mmol), from Preparation 14 (b), in 48% aqueous HBr (50 mL), was heated to reflux for 12 h. The reaction was concentrated and toluene was used to azeotropically remove residual H 2 O. The resulting light brown solid was placed under high vacuum overnight and used directly.

b) Methyl (±) -6-Bromo-2-oxo-1,2,3,4-tetrahydro-1H-1,8-naphthyridine-3-carboxylate

Dimethyl malonate (11.87 g, 89.9 mmol) was added to a solution of sodium methoxide (20.57 mL, 25 wt% in CH3OH) in CH3OH (75 mL). After 30 min, the 2-amino-5-bromo-3- (bromomethyl) pyridine hydrobromide prepared above was added to the methoxide solution and the reaction was stirred at room temperature overnight. The reaction slurry was concentrated to dryness under reduced pressure then suspended in 1: 1 H 2 O / Et 2 O. The remaining solids were filtered and washed with H 2 O then hexane to obtain the title compound (4.08 g, 58%) as a white solid after drying.

MS (ES) m / e 286 (M + H)⁺.

c) 6-Bromo-3,4-dihydro-1H-1,8-naphthyridin-2-one

To a solution of methyl (±) -6-bromo-2-oxo-1,2,3,4-tetrahydro-1H-1,8-naphthyridine-3-carboxylate (2.00 g, 7.0 mmol) in CH3OH ( 75 mL) was added 1.0 M NaOH (30 mL). The reaction was heated to reflux for 4 h and then cooled to room temperature. The reaction was neutralized with 1.0 M HCl (30 mL) then heated to reflux overnight. The reaction slurry was concentrated to dryness and the residues were suspended in 95: 5 CHCl3 / CH3OH. The solids were removed by filtration and the filtrate was concentrated to provide the title compound (1.40 g, 88%) as an off-white solid.

MS (ES) m / e 228 (M + H)⁺.

Production 17

Preparation of (E) -3- [6-amino-5 - [(2-hydroxyethylamino) carbonyl] pyridin-3-yl] acrylic acid

a) 2-Amino-5-bromo-N- (2-hydroxyethyl) nicotinamide

EDC (2.91 g, 15.2 mmol) was added to a solution of 2-amino-5-bromonicotinic acid (3.00 g, 13.8 mmol), ethanolamine (0.93 g, 15.2 mmol), HOBt H 2 O (2.05 g, 15.2 mmol) and diisopropylethylamine (2.64 mL, 15.2 mmol) in DMF (50 mL) at room temperature and the reaction solution was stirred overnight. The reaction mixture was poured into H 2 O (200 mL) and the resulting mixture was extracted with EtOAc (2 x 200 mL). The combined organic extracts were washed with H 2 O and brine then dried over Na 2 SO 4. Concentration of the organic extracts gave the title compound as a yellow solid which was used without further purification.

MS (ES) m / e 261 (M + H)⁺.

b) benzyl (E) -3- [6-Amino-5 - [(2-hydroxyethylamino) carbonyl] pyridin-3-yl] acrylate

Following the procedure for Preparation 2 (a) except that 2-amino-5-bromo-N- (2-hydroxyethyl) nicotinamide (2.70 g, 10.4 mmol) was used instead of 2-amino-5- bromopyridine, the title compound (2.67 g, 75%) was prepared as an off-white solid.

MS (ES) m / e 342 (M + H)⁺.

c) (E) -3- [6-Amino-5 - [(2-hydroxyethylamino) carbonyl] pyridin-3-yl] acrylic acid

Following the procedure for Preparation 2 (b) except that benzyl (E) -3- [6-amino-5 - [(2-hydroxyethylamino) carbonyl] pyridin-3-yl] acrylate was used (2.67 g , 7.8 mmol) instead of benzyl (E) -3- (6-aminopyridin-3-yl) acrylate, the title compound (1.37 g, 70%) was prepared as an off-white solid.

MS (ES) m / e 252 (M + H)⁺.

PL 201 627 B1

Production 18

Preparation of 6-bromo-3-methyl-3,4-dihydro-1H-pyrido [2,3-d] pyrimidin-2-one

a) 2-Amino-5-bromo-3- (methylaminomethyl) pyridine

A solution of 2-amino-5-bromo-3- (hydroxymethyl) pyridine (5.00 g, 24.6 mmol) from Preparation 14 (b) in 48% aqueous HBr (50 mL) was heated to reflux for 12 h. The reaction was concentrated and toluene was used to azeotropically remove residual H 2 O. The resulting light brown solid was placed under high vacuum overnight and used directly. A solution of 2-amino-3- (bromomethyl) -5-bromopyridine hydrobromide (prepared above) in 40% aqueous methylamine (50 mL) and THF (50 mL) was stirred at room temperature overnight in a pressure bottle. The reaction solution was concentrated and extracted with EtOAc (2 x 100 mL). The combined organic phases were washed with H 2 O, dried over Na 2 SO 4, and concentrated. Purification on silica gel gave the title compound (4.25 g, 80%) as yellow oil.

MS (ES) m / e 217 (M + H)⁺.

b) 6-Bromo-3-methyl-3,4-dihydro-1H-pyrido [2,3-d] pyrimidin-2-one

To a solution of dimethyl carbonate (2.14 g, 23.7 mmol) and sodium methoxide (1.0 mL, 4.5 mmol, 25 wt% in CH3OH) in CH3OH (25 mL) was added 2-amino-5-bromo -3- (methylaminomethyl) pyridine (1.0 g, 4.62 mmol). The reaction was heated at 50 ° C overnight, diluted with H 2 O (1 mL) and concentrated. Toluene was added to the reaction residue and the contents were heated to reflux for 12 h under a Dean-Stark separator. The reaction was cooled to room temperature, diluted with EtOAc, and washed with H20. Purification on silica gel (9: 1 CHCl3 / CH3OH containing 5% NH4OH) gave the title compound (0.75 g, 67%) as an off-white solid.

MS (ES) m / e 243 (M + H)⁺.

Production 19

Preparation of 4-methyl-5- (methylaminomethyl) -4H-thieno [3,2-b] pyrrole

a) Ethyl 4-methyl-4H-thieno [3,2-b] pyrrole-5-carboxylate

Following the procedure for Preparation 1 (a) except that ethyl 4H-thieno [3,2-b] pyrrole-5-carboxylate (1.30 g, 6.7 mmol, see J. Het. Chem. 1984, 21, 215-217) in place of ethyl indole-2-carboxate, provided the title compound (1.35 g, 97%) as yellow solid.

MS (ES) m / e 210 (M + H)⁺.

b) N, 4-Dimethyl-4H-thieno [3,2-b] pyrrole-5-carboxamide

Following the procedure from Preparation 1 (b) except that ethyl 4H-thieno [3,2-b] pyrrole-5-carboxylate (1.35 g, 6.5 mmol) was used instead of 1- ethyl methylindole-2-carboxylate was obtained, the title compound (1.19 g, 95%) was prepared as a yellow solid.

MS (ES) m / e 195 (M + H)⁺.

c) 4-Methyl-5- (methylaminomethyl) -4H-thieno [3,2-b] pyrrole

Following the procedure from Preparation 1 (c) except that N, 4-dimethyl-4H-thieno [3,2-b] pyrrole-5-carboxamide (0.70 g, 3.6 mmol) was used in place of N , 1-dimethylindole-2-carboxamide, gave the title compound (0.60 g, 92%) as yellow oil.

MS (ES) m / e 181 (M + H)⁺.

Production 20

Preparation of 3-methyl-2- (methylaminomethyl) indene hydrochloride

a) N, 3-Dimethylindene-2-carboxamide

EDC (1.53 g, 0.01 mol) was added to a solution of 3-methyl-2-inden-2-carboxylic acid (1.91 g, 0.01 mol), methylamine hydrochloride (0.675 g, 0.01 mol) , HOBt · H 2 O (1.53 g, 0.01 mol) and triethylamine (4.0 mL, 0.028 mol) in anhydrous DMF (80 mL) at room temperature. The reaction was stirred overnight and then concentrated under reduced pressure. The residue was diluted with 5% NaHCO3 and the resulting white precipitate was collected, washed with water and dried at 50 ° C in a vacuum oven to give the title compound (1.6 g, 86%) as a white solid.

MS (ES) m / e 188.2 (M + H)⁺.

b) 3-Methyl-2- (methylaminomethyl) indene hydrochloride

The flame dried flask was charged with anhydrous THF (15 mL) followed by solid lithium aluminum hydride (760 mg, 0.02 mol) at 0 ° C. The mixture was stirred for 15 min, then a solution of N, 3-dimethylindene-2-carboxamide (1.5 g, 0.008 mol) in anhydrous THF (20 mL) was added dropwise. When the addition was complete, the reaction was heated under gentle reflux for 30 h, then cooled in ice and quenched by the addition of H 2 O (1.4 mL) and NaF (2.5 g, 0.06 mol). Mixed30

The reaction mixture was stirred for 40 min then filtered through celite^® and the filter pad was washed with THF. The filtrate was dried over K2CO3, filtered and concentrated to an oil which was dissolved in anhydrous diethyl ether and treated with 4M HCl in diethyl ether. The precipitated light beige solid was collected by filtration and washed with diethyl ether. Drying at 50 ° C in a vacuum oven gave the title compound (1.05 g, 80.7%) as a light beige solid.

MS (ES) m / e 174.2 (M + H)⁺.

Production 21

Preparation of 2- (methylaminomethyl) indene hydrochloride

a) N-Methylindene-2-carboxamide

According to the procedure of Preparation 20 (a), except that 2-inden-carboxylic acid was used in place of 3-methyl-2-inden-2-carboxylic acid, the title compound was obtained as a white crystalline solid (1.45 g, 83.3%).

MS (ES) m / e 174.2 (M + H)⁺.

b) 2- (methylaminomethyl) indene hydrochloride

Following the procedure of Preparation 20 (b), except that N-methylindene-2-carboxamide was used in place of N, 3-dimethylindene-2-carboxamide, the title compound was obtained as an off-white solid (0.685 g, 87.6%) .

MS (ES) m / e 160.0 (M + H)⁺.

Production 22

Preparation of 4-methoxy-1-methyl-2- (methylaminomethyl) -1H-indole hydrochloride

a) Methyl 4-methoxy-1-methyl-1H-indole-2-carboxylate

NaH (60% dispersion in mineral oil, 0.3 g, 7.3 mmol) was washed with hexane then suspended in anhydrous DMF (16 mL). The mixture was cooled to 0 ° C and methyl 4-methoxy-1H-indole-2-carboxylate (1.0 g, 4.87 mmol) was added. The mixture was stirred under argon for 10 min, then MeI (1.3 mL, 20 mmol) was added and the thick suspension was stirred at room temperature for 2.5 h. The reaction was quenched with 10% NH4Cl (2 mL) and concentrated. The residue was partitioned between H 2 O and Et 2 O and the organic layer was dried over MgSO 4 and concentrated to give the title compound (1.03 g, 96%) as a white solid.

MS (ES) m / e 220.2 (M + H)⁺.

b) N, 1-Dimethyl-4-methoxy-1H-indole-2-carboxamide

A solution of methyl 4-methoxy-1-methyl-1H-indole-2-carboxylate (1.03 g, 4.7 mmol) in 2M methylamine in methanol (40 mL) was sealed in a pressure bottle and heated at 55-60 ° C for 60 h. Concentration under reduced pressure gave the title compound (1.05 g, quantitative) as a white solid.

MS (ES) m / e 219.2 (M + H)⁺.

c) 4-methoxy-1-methyl-2- (methylaminomethyl) -1H-indole hydrochloride

According to the procedure for preparation of 20 (b), except that N, 1-dimethyl-4-methoxy-1H-indole-2-carboxamide was used instead of N, 3-dimethylindene-2-carboxamide, the title compound was obtained as an off-white substance solid (0.72 g, 75%).

MS (ES) m / e 205.2 (M + H)⁺.

Production 23

Preparation of 1,4-dimethyl-2- (methylaminomethyl) -1H-indole hydrochloride

a) 1,4-Dimethyl-1H-indole-2-carboxylic acid

A solution of 1,4-dimethyl-1H-indole (0.9 g, 6.2 mmol) in anhydrous Et2O (20 mL) was treated with 2.5M n-BuLi in hexane (5.0 mL, 12 mmol) and the reaction was heated in reflux for 15 h. The dark reaction mixture was poured onto a slurry of excess crushed dry ice in Et2O and the mixture was allowed to stand for 1 h. Water (10 mL) was added, the layers were separated and the aqueous layer was filtered through Celite^®. The clear filtrate was acidified by the addition of 2.0N HCl to pH 2 and the precipitate was collected and dried to afford the title compound (0.29 g, 26.4%) as an off-white solid.

MS (ES) m / e 190.2 (M + H)⁺.

b) N, 1,4-Trimethyl-1H-indole-2-carboxamide

Following the procedure for Preparation 20 (a) except that 1,4-dimethyl-1H-indole-2-carboxylic acid was used in place of 3-methyl-2-indene-2-carboxylic acid, provided the title compound (0.184 g, 91%).

MS (ES) m / e 203.2 (M + H)⁺.

PL 201 627 B1

c) 1,4-dimethyl-2- (methylaminomethyl) -1H-indole hydrochloride

Following the procedure for Preparation 20 (b) except that N, 1,4-trimethyl-1H-indole-2-carboxamide was used in place of N, 3-dimethylindene-2-carboxamide, provided the title compound (0.13 g , 65%).

MS (ES) m / e 189.2 (M + H)⁺.

Production 24

Preparation of 2- (Cyclopropylamino) -1-methyl-1H-indole

a) 2- (Cyclopropylamino) -1-methyl-1H-indole

NaBH3CN was added to a solution of 1-methylindole-2-carboxaldehyde (1.5 g, 10 mmol), cyclopropylamine (1.14 g, 20 mmol) and glacial acetic acid (0.6 mL, 10 mmol) in MeOH (30 mL) (0.69 g, 11 mmol). The reaction was stirred at room temperature overnight then concentrated under reduced pressure. The residue was diluted with 10% NaOH and extracted with CH2Cl2. The combined organic extracts were washed with brine, dried over MgSO4, and concentrated. Flash chromatography on silica gel (3% MeOH / CH2Cl2) gave the title compound (1.3 g, 65%) as a semi-solid.

MS (ES) m / e 201 (M + H)⁺.

Generation 25

Preparation of 5-fluoro-2- (methylaminomethyl) -1H-indole

a) Ethyl 5-fluoro-1-methyl-1H-indole-2-carboxylate

Following the procedure from Preparation 1 (a) except that ethyl 5-fluoro-indole-2-carboxylate was used in place of ethyl indole-2-carboxylate, the title compound (3.3 g, 100%) was obtained as a white solid .

MS (ES) m / e 222 (M + H)⁺.

b) N, 1-Dimethyl-5-fluoro-1H-indole-2-carboxamide

Following the procedure for Preparation 1 (b) except that ethyl 5-fluoro-1-methyl-1H-indole-2-carboxylate was used in place of ethyl 1-methyl-1H-indole-2-carboxylate, the title compound ( 2.1 g, 68%) as a white solid.

MS (ES) m / e 207 (M + H)⁺.

c) 5-Fluoro-2- (methylaminomethyl) -1H-indole

Prepared following the procedure for preparing 1 (c) except that N, 1-dimethyl-5-fluoro-1H-indole-2-carboxamide was used in place of N, 1-dimethyl-1H-indole-2-carboxamide, title (1.5 g, 78%) as a white solid.

MS (ES) m / e 193 (M + H)⁺.

Production 26

Preparation of 3- (methylaminomethyl) quinoline

a) 3- (Methylaminomethyl) quinoline

A solution of 3-quinolinecarboxaldehyde (1.5 g, 10 mmol), 2.0 M CH3NH2 / MeOH (10 mL, 20 mmol), glacial AcOH (0.6 mL, 10 mmol) and NaBH3CN (0.35 g, 11 mmol) ) in MeOH (20 mL) was stirred at room temperature overnight then concentrated under reduced pressure. The residue was diluted with 5% NaOH and extracted with CH2Cl2. The combined organic extracts were washed with brine, dried over MgSO4, and concentrated. Flash chromatography on silica gel (10% MeOH / CH2Cl2) gave the title compound (0.83 g, 24%) as yellowish viscous oil.

MS (ES) m / e 173 (M + H)⁺.

Production 27

Preparation of 2- (methylaminomethyl) benzofuran

a) N-Methylbenzofuran-2-carboxamide

To a solution of 2-benzofurancarboxylic acid (1.62 g, 10 mmol), methylamine hydrochloride (0.79 g, 11 mmol), methylamine (3.1 mL, 22 mmol), and HOBtH2O (1.5 g, 11 mmol) mmol) in DMF (30 mL) was added EDC (2.1 g, 11 mmol). The reaction was stirred overnight then concentrated under reduced pressure. The residue was diluted with 5% NaHCO3 and extracted with CH2Cl2. The combined organic extracts were washed with brine, dried over MgSO4, and concentrated. Flash chromatography on silica gel (3% MeOH / CH2Cl2) gave the title compound (1.75 g, 100%) as a white solid.

MS (ES) m / e 176 (M + H)⁺.

b) 2- (Methylaminomethyl) benzofuran

N-methylbenzofuran-2-carboxamide (1.75 g, 10 mmol) was added to a 1M BH3 / THF solution (30 mL, 30 mmol) at 0 ° C. The reaction mixture was allowed to warm to temperature

Room temperature, then refluxed overnight. The reaction was cooled to 0 ° C and excess methanol was added. The resulting solution was concentrated under reduced pressure, and the residue was purified by flash chromatography on silica gel (3% MeOH / CH2Cl2). The title compound was obtained (0.2 g, 12%) as a white solid.

MS (ES) m / e 162 (M + H)⁺.

Generation 28

Preparation of 1-methyl-2- (propylaminomethyl) -1H-indole

a) 1-Methyl-N-cyclopropylindole-2-carboxamide

Following the procedure for Preparation 27 (a), except that 1-methyl-1H-indole-2-carboxylic acid (3.5 g, 20 mmol) was used in place of 2-benzofurancarboxylic acid and cyclopropylamine was used in place of methylamine hydrochloride, the compound was prepared title (2.1 g, 49%) as a white solid.

MS (ES) m / e 215 (M + H)⁺.

b) 1-Methyl-2- (propylaminomethyl) -1H-indole

To a solution of 1-methyl-N-cyclopropylindole-2-carboxamide (2.1 g, 9.8 mmol) in dry THF (40 mL) was added dropwise a solution of 1.0 M LiAlH4 in THF (2.2 mL, 22 mmol) . The reaction mixture was refluxed overnight then cooled and quenched with 10% NaOH. The mixture was filtered and the filtrate was concentrated under reduced pressure. Flash chromatography on silica gel (5% MeOH / CH2Cl2) gave the title compound (0.65 g, 33%) as a viscous oil.

MS (ES) m / e 203 (M + H)⁺.

Generation 29

Preparation of 5-bromo-2- (methylamino) pyridine and 5-bromo-2- (dimethylamino) pyridine

a) 5-Bromo-2- (methylamino) pyridine and 5-bromo-2- (dimethylamino) pyridine

To a suspension of NaH (60% dispersion in mineral oil, 0.44 g, 11 mmol) in dry DMF (40 mL) was added solid 2-amino-5-bromopyridine (1.73 g, 10 mmol) portionwise over 5-10 min. Gas evolution was allowed to stop between the additions. The resulting amber mixture was stirred for 15 min before all methyl iodide (0.61 mL, 10 mmol) was added all at once. The reaction mixture was stirred at room temperature overnight then concentrated under reduced pressure. The residue was diluted with 5% NH4Cl (30 mL) and the mixture was extracted with CH2Cl2. The combined organic extracts were washed with brine, dried (MgSO4) and concentrated. Flash chromatography on silica gel (3% MeOH / CH2Cl2) separated the products.

5-Bromo-2- (methylamino) pyridine (0.60 g, 32%) was obtained as a semi-solid: TLC (3% MeOH / CH2Cl2) Rf 0.35; MS (ES) m / e 187 (M + H)⁺.

5-Bromo-2- (dimethylamino) pyridine (0.70 g, 34%) was obtained as a semi-solid: TLC (3% MeOH / CH2Cl2) R f 0.77; MS (ES) m / e 201 (M + H)⁺.

Generation 30

Preparation of (E) -3- [6- (methylamino) pyridin-3-yl] acrylic acid

a) benzyl (E) -3- [6-methylamino) pyridin-3-yl) acrylate

Following the procedure of Preparation 2 (a), except that 5-bromo-2- (methylamino) pyridine was used in place of 2-amino-5-bromopyridine, the title compound (0.52 g, 60%) was prepared as a white material constant.

MS (ES) m / e 269 (M + H)⁺.

b) (E) -3- [6- (methylamino) pyridin-3-yl] acrylic acid

Following the procedure for Preparation 2 (b), except that benzyl (E) -3- [6- (methylamino) pyridin-3-yl] acrylate was used instead of (E) -3- (6-aminopyridin-3- benzyl acrylate, the title compound (0.15 g, 43%) was prepared as a white solid.

MS (ES) m / e 179 (M + H)⁺.

Production 31

Preparation of (E) -3- [6- (dimethylamino) pyridin-3-yl] acrylic acid

a) benzyl (E) -3- [6- (dimethylamino) pyridin-3-yl] acrylate

Following the procedure of Preparation 2 (a) except that 5-bromo-2- (dimethylamino) pyridine was used in place of 2-amino-5-bromopyridine, the title compound (0.82 g, 84%) was prepared as a white material constant.

MS (ES) m / e 283 (M + H)⁺.

PL 201 627 B1

b) (E) -3- [6- (dimethylamino) pyridin-3-yl] acrylic acid

Following the procedure for Preparation 2 (b), except that benzyl (E) -3- [6- (dimethylamino) pyridin-3-yl] acrylate was used instead of (E) -3- (6-aminopyridin-3- benzyl acrylate, the title compound (0.20 g, 36%) was prepared as a white solid.

MS (ES) m / e 193 (M + H)⁺.

Generation 32

Preparation of (E) -3- (6-methylpyridin-3-yl) acrylic acid

a) benzyl (E) -3- (6-methylpyridin-3-yl) acrylate

Following the procedure for Preparation 2 (a) except that 5-bromo-2-methylpyridine was used in place of 2-amino-5-bromopyridine, the title compound (0.85 g, 34%) was obtained as a white solid.

MS (ES) m / e 253 (M + H)⁺.

b) (E) -3- (6-methylpyridin-3-yl) acrylic acid

Following the procedure for the preparation of 2 (b), except that benzyl (E) -3- (6-methylpyridin-3-yl) acrylate was used instead of (E) -3- (6-aminopyridin-3-yl) acrylate benzyl, the title compound (0.18 g, 33%) was prepared as a white solid

MS (ES) m / e 164 (M + H)⁺.

Generation 33

Preparation of 2- (methylaminomethyl) -1H-indole

a) N-Methyl-1H-indole-2-carboxamide

A suspension of ethyl indole-2-carboxylate (25.30 g, 133.7 mmol) in 40% aqueous CH3NH2 (400 mL) was stirred at room temperature. The flask was tightly capped to retain the material inside the flask. As the reaction proceeded, the product began to precipitate. The reaction was stirred at room temperature for 3 days then concentrated to remove approximately 200 mL of solvent. The residue was diluted with H2O (500 mL) and the solid was collected by filtration and washed with H2O. Drying in vacuo gave the title compound (21.50 g, 92%) as a light yellow solid.

MS (ES) m / e 175 (M + H)⁺.

b) 2- (Methylaminomethyl) -1H-indole

A solution of LiAlH4 in THF (1.0 M, 250 mL, 250 mmol) was slowly added via syringe to a solution of N-methyl-1H-indole-2-carboxamide (21.50 g, 12.34 mmol) in anhydrous THF (100 mL) . Gas was evolved during the addition of the first 50 mL of LiAlH4 solution. When the addition was complete, the resulting pale yellow solution was heated to gentle reflux. After 23 h, the reaction was cooled in ice and quenched by the sequential dropwise addition of H 2 O (9.5 mL), 1.0 N NaOH (20 mL), and H 2 O (28.5 mL). The mixture was stirred for 15 min, then filtered through Celite^® and the filter pad was washed thoroughly with THF. The filtrate was concentrated and the residue was flash chromatographed on silica gel (10% MeOH / CHCl3 containing 0.5% conc. NH4OH). The title compound (10.10 g, 51%) was obtained as a light yellow oil.

MS (ES) m / e 161 (M + H)⁺.

Generation 34

Preparation of 1-ethyl-2- (methylaminomethyl) -1H-indole

a) 2- [N- (Benzyloxycarbonyl) -N-methylaminomethyl] -1H-indole

N- (Benzyloxycarbonyloxy) succinimide (17.10 g, 68.6 mmol) was added to a solution of 2- (methylamino-methyl) -1H-indole (10.00 g, 62.4 mmol) for the preparation of 33 and triethylamine (9.60 mL, 68.6 mmol) in DMF (100 mL) at room temperature. The reaction was stirred overnight then concentrated under reduced pressure. The residue was diluted with water, and the mixture was extracted with ethyl acetate. The combined extracts were dried over K2CO3 and concentrated. Flash chromatography on silica gel (20% ethyl acetate / hexane) gave the title compound (14.80 g, 80%) as an off-white solid.

MS (ES) m / e 295 (M + H)⁺.

b) 2- [N- (Benzyloxycarbonyl) -N-methylaminomethyl] -1-ethyl-1H-indole

NaH (60% dispersion in mineral oil, 0.25 g, 7.1 mmol) was added portionwise, allowing gas evolution, to a solution of 2- [N- (benzyloxycarbonyl) -N-methylaminomethyl] -1H-indole (1.40 g, 4.75 mmol) in DMF (35 mL) at 0 ° C. When the NaH addition was complete, ethyl iodide (0.42 mL, 5.2 mmol) was added at 0 ° C. The reaction was stirred at 0 ° C for

Minutes and then at room temperature overnight. The reaction was diluted with water and extracted with ethyl acetate. The combined extracts were dried over K2CO3 and concentrated to give the title compound (1.30 g, 87%) as an orange solid.

MS (ES) m / e 323 (M + H)⁺. e) 1-Ethyl-2- (methylaminomethyl) -1H-indole

2- [N- (Benzyloxycarbonyl) -N-methylaminomethyl] -1-ethyl-1H-indole (1.30 g, 4.0 mmol) was added to the Pearlman's catalyst slurry (about 0.30 g in MeOH at room temperature in a Parr bottle The reaction was placed under 50 psi of H 2 and shaken for 8 h.

The mixture was filtered through celite^® and the filter pad was washed with MeOH. The filtrate was concentrated to provide the title compound (0.75 g, 100%) as a light yellow solid.

MS (ES) m / e 189 (M + H)⁺.

Generation 35

Preparation of 1-methyl-3- (methylaminomethyl) -1H-indole (Method A)

a) Methyl 1-methyl-1H-indole-3-carboxylate

NaH (60% dispersion in mineral oil, 8.56 g, 214.0 mmol) was added portionwise, allowing gas evolution, to a solution of methyl 1H-indole-3-carboxylate (25 g, 142.7 mmol) in DMF (350 mL) at 0 ° C.

When the NaH addition was complete, methyl iodide (44.4 mL, 713.5 mmol) was added at 0 ° C. The reaction was stirred at 0 ° C for 15 minutes then at room temperature overnight. The reaction was diluted with water and extracted with ethyl acetate.

The combined extracts were dried over K2CO3 and concentrated to afford the title compound (26.00 g, 96%) as an orange solid.

MS (ES) m / e 190 (M + H)⁺.

b) N, 1-Dimethyl-1H-indole-3-carboxamide

A suspension of methyl 1H-indole-3-carboxylate (4.30 g, 22.74 mmol) in 40% aqueous CH3NH2 (400 mL) was stirred at room temperature.

The flask was tightly capped to retain the material inside the flask. As the reaction proceeded, the product began to precipitate.

The reaction was stirred at room temperature for 3 days then concentrated to remove approximately 200 mL of solvent. The residue was diluted with H2O (500 mL) and the solid was collected by filtration and washed with H2O.

Flash chromatography on silica gel (ethyl acetate) gave the title compound (2.4 g, 56%) as a white solid.

MS (ES) m / e 189 (M + H)⁺.

c) 1-Methyl-3- (methylaminomethyl) -1H-indole

A solution of LiAlH4 in THF (1.0M, 5.20 mL, 5.2 mmol) was slowly added by syringe to a solution of N, 1-dimethyl-1H-indole-3-carboxamide (0.50 g, 2.6 mmol) in anhydrous THF (15 mL). Gas was evolved during the addition of the first 2 mL of LiAlH4 solution. When the addition was complete, the resulting pale yellow solution was heated to gentle reflux.

After 23 h, the reaction was cooled in ice and quenched by the sequential dropwise addition of H 2 O (0.5 mL), 1.0 N NaOH (0.5 mL), and H 2 O (0.5 mL). The mixture was stirred for 15 min, then filtered through Celite^® and the filter pad was washed thoroughly with THF.

The filtrate was concentrated and the residue was flash chromatographed on silica gel (10% MeOH / CHCl3 containing 0.5% conc NH4OH) to give the title compound (0.30 g, 67%) as a light yellow oil.

MS (ES) m / e 175 (M + H)⁺.

Generation 36

Preparation of 1-methyl-3- (methylaminomethyl) -1H-indole (Method B)

To a solution of 1-methylindole-3-carboxaldehyde (10.0 g, 62.8 mmol) in MeOH (100 mL) was added a solution of 2.0 M CH3NH2 in Moss (126 mL, 252.0 mmol). The reaction was stirred at room temperature for 2 h, then concentrated to a light yellow oil. This oil was dissolved in EtOH (300 mL) and NaBH4 (2.38 g, 62.8 mmol) was added. After 2 h, the reaction was concentrated to a slurry and dissolved in 1.0 N NaOH (75 mL). The aqueous solution was extracted (2 x 200 mL) and the combined organic fractions were dried over Na2SO4 and concentrated. Flash chromatography on silica gel (9: 1 CHCl3 / MeOH containing 5% NH4OH) and drying under high vacuum gave the title compound (10.1 g, 92%) as a pale yellow oil.

MS (ES) m / e 175 (M + H)⁺.

PL 201 627 B1

Production 37

Preparation of HCl salt of (E) -3- (6-aminopyridin-3-yl) -2-methylacrylic acid and HCl salt of acid

2- (6-aminopyridin-3-ylmethyl) acrylic

a) ethyl (E) -3- (6-aminopyridin-3-yl) -2-methylacrylate and ethyl 2- (6-aminopyridin-3-ylmethyl) acrylate

To a stirred solution of 2-amino-5-bromopyridine (25 g, 140 mmol) in propionitrile (150 mL) was added ethyl methacrylate (50 mL, 400 mmol), DIEA (50 mL, 287 mmol), palladium (II) acetate (1 , 57 g, 7 mmol) and tri-o-tolylphosphine (4.3 g, 14 mmol). The reaction was purged with argon and refluxed for 6 h, then cooled to room temperature and concentrated to dryness under reduced pressure. The residue was dissolved in 80% ethyl acetate / hexane (100 mL) and the solution was filtered through a pad of silica gel, eluting with 80% ethyl acetate / hexane (400 mL) until all product was eluted. The yellowish filtrate was concentrated under reduced pressure and the residue was dissolved in a small volume of 1: 1 Et2O / petroleum ether. The resulting precipitate was collected and dried under reduced pressure to obtain ethyl (E) -3- (6-aminopyridin-3-yl) -2-methylacrylate (10.77 g, 37%) as a pale yellow solid.

LCMS (ES) m / e 207.0 (M + H)⁺;

¹H NMR (300 MHz, CDCl3) δ 8.05 (d, J = 1.7 Hz, 1H), 7.63 (dd, 1H), 7.48 (s, 1H), 6.75 (d, J = 8.8 Hz, 1H), 5.79 (br s, 2H), 4.26 (q, 2H), 2.10 (s, 3H), 1.34 (t, 3H).

The filtrate was concentrated to dryness and purified by silica gel flash chromatography (4: 1 ethyl acetate / hexane) to give additional ethyl (E) -3- (6-aminopyridin-3-yl) -2-methylacrylate (0.87 g , 3%) and ethyl 2- (6-aminopyridin-3-ylmethyl) acrylate (5.77 g, 20%) as yellow oil.

LCMS (ES) m / e 207.0 (M + H)⁺;

¹H NMR (300 MHz, CDCl 3) δ 7.86 (d, J = 2.1 Hz, 1H), 7.32 (dd, 1H), 6.53 (d, J = 8.5 Hz, 1H), 6.21 (d, J = 1.8 Hz, 1H), 5.48 (d, J = 1.4 Hz, 1H), 4.17 (q, 2H), 3.47 (s, 2H), 1.27 (t, 3H).

b) HCl salt of (E) -3- (6-aminopyridin-3-yl) -2-methylacrylic acid

HOAc (25 mL) and concentrated HCl (25 mL) were added to ethyl (E) -3- (6-aminopyridin-3-yl) -2-methylacrylate (5.0 g, 24.2 mmol). The reaction was stirred and heated at 100 ° C for 6 h, cooled to room temperature and concentrated to dryness. The residue was triturated with Et2O, filtered and dried under reduced pressure to give the title compound (5.5 g, quantitative) as a white solid.

LCMS (ES) m / e 179.0 (M + H)⁺;

¹H NMR (300 MHz, DMSO-d6) δ 8.47 (br s, 2H), 8.16 (d, J = 1.7 Hz, 1H), 8.08 (dd, 1H), 7.42 ( s, 1H), 7.08 (d, J = 9.3 Hz, 1H), 2.01 (s, 3H).

c) HCl salt of 2- (6-aminopyridin-3-ylmethyl) acrylic acid

Following the procedure from Preparation 37 (b), except that ethyl 2- (6-aminopyridin-3-ylmethyl) acrylate (3.1 g, 15 mmol) was used instead of (E) -3- (6-aminopyridin- Ethyl 3-yl) -2-methylacrylate to give the title compound (3.0 g, 93%) as a white solid.

LCMS (ES) m / e 179.0 (M + H)⁺;

¹H NMR (300 MHz, DMSO-d6) δ 8.10 (br s, 2H), 7.79 (dd, 1H), 7.78 (s, 1H), 7.00 (d, J = 9.7 Hz, 1H), 6.15 (d, J = 1.2 Hz, 1H), 5.67 (d, J = 1.2 Hz, 1H), 3.45 (s, 2H).

Generation 38

Preparation of 2- (methylaminomethyl) naphthalene

For a mixed solution of 40 wt. methylamine in H 2 O (50 mL, 581 mmol) in THF (50 mL) at 0 ° C was added 2- (bromomethyl) naphthalene (10 g, 43 mmol) in one portion. The reaction was allowed to warm to room temperature and stirred for 16 h then concentrated under reduced pressure. The residue was dissolved in Et2O and washed with 1.0 N NaOH, then brine, dried (Na2SO4) and concentrated to dryness. Purification by flash chromatography on silica gel (98: 2 to 9: 1 CHCl3 / methanol containing 5% NH4OH) gave the title compound (3.95 g, 54%) as clear oil.

¹H NMR (400 MHz, CDCl3) δ 7.85 (m, 3H), 7.79 (s, 1H), 7.49 (m, 3H), 3.94 (s, 2H), 2.53 (s , 3H).

Generation 39

Preparation of the HCl salt of (E) -3- (6-amino-4-methyl-pyridin-3-yl) acrylic acid

a) 2-Amino-5-bromo-4-methylpyridine

To a stirred solution of 2-amino-4-methylpyridine (22 g, 203 mmol) in 48% HBr (200 mL) at 70 ° C was added a solution of 15% H 2 O 2 in H 2 O (60 mL) dropwise over 60 minutes. Reaction

The material became somewhat exothermic and the oil bath was removed after 15 minutes. The reaction was stirred for an additional 1 h, then poured onto ice (approximately 500 mL). The clear solution was adjusted to pH 4-5 with solid Na2CO3 (80 g, 755 mmol) and the resulting thick white suspension was filtered. The filter pad was washed with a little H 2 O and pressed dry. Drying in vacuo gave a 2: 3 mixture of 2-amino-5-bromo-4-methylpyridine and 2-amino-3,5-dibromo-4-methylpyridine (27.08 g). Flash silica gel chromatography (50% ethyl acetate / hexane then ethyl acetate) gave the title compound (12.11 g, 32%) as a white solid.

LCMS (ES) m / e 187.2 (M + H)⁺;

¹H NMR (400 MHz, DMSO-d6) δ 7.92 (s, 1H), 6.41 (s, 1H), 6.03 (br s, 2H), 2.17 (s, 3H).

b) ethyl (E) -3- (6-Amino-4-methylpyridin-3-yl) acrylate

To a stirred solution of 2-amino-5-bromo-4-methylpyridine (10 g, 54 mmol) in propionitrile (50 mL) was added ethyl acrylate (17 mL, 157 mmol), DIEA (19 mL, 106 mmol), palladium acetate ( II) (0.61 g, 2.7 mmol) and tri-o-tolylphosphine (1.64 g, 5.4 mmol). The reaction was purged with argon and refluxed for 6 h, then cooled to room temperature and concentrated to dryness under reduced pressure. The obtained residue was dissolved in ethyl acetate and filtered through a pad of silica gel. The filtrate was concentrated and the residue was triturated with 1: 1 Et2O / petroleum ether (50 mL), filtered and dried under reduced pressure to provide the title compound (6.50 g, 59%) as a pale yellow solid.

¹H NMR (400 MHz, DMSO-d6) δ 8.31 (s, 1H), 7.66 (d, J = 16.0 Hz, 1H), 6.40 (br s, 2H), 6.32 ( d, J = 16.0 Hz, 1H), 6.28 (s, 1H), 4.15 (q, 2H), 2.24 (s, 3H), 1.24 (t, 3H).

c) HCl salt of (E) -3- (6-amino-4-methyl-pyridin-3-yl) -acrylic acid

HOAc (15 mL) and concentrated HCl (15 mL) were added to ethyl (E) -3- (6-amino-4-methylpyridin-3-yl) acrylate (1.50 g, 7.3 mmol). The solution was stirred at 100 ° C for 10 h, cooled to room temperature, and concentrated to dryness. Trituration with Et2O, filtration and drying in vacuo gave the title compound (1.65 g, quantitative) as a white solid.

LCMS (ES) m / e 179.2 (M + H)⁺;

¹H NMR (400 MHz, DMSO-d6) δ 8.37 (s, 1H), 8.28 (br s, 3H), 7.51 (d, J = 16.0 Hz, 1H), 6.86 ( s, 1H), 6.46 (d, J = 16.0 Hz, 1H), 2.41 (s, 3H).

Generation 40

Preparation of 1,3-dimethyl-2- (methylaminomethyl) -1H-indole

a) 1,3-Dimethyl-1H-indole

To a stirred solution of 3-methylindole (15.0 g, 114 mmol) in dry DMF (200 mL) was added NaH (60% oil dispersion, 5.0 g, 125 mmol) portionwise. Gas evolution was observed. The mixture was stirred for 30 min, then iodomethane (8 mL, 129 mmol) was added in one portion. The reaction became exothermic and was cooled in an ice bath. After 16 h at room temperature, the reaction was concentrated under reduced pressure and the residue was dissolved in ethyl acetate. The solution was washed with H 2 O then brine, dried (MgSO 4) and concentrated to dryness. Purification by ball distillation under reduced pressure (bp 88-92 ° C, 0.5 mm of Hg) provided the title compound (16.10 g, 97%) as pale yellow oil.

¹H NMR (400 MHz, CDCl 3) δ 7.47 (d, J = 7.9 Hz, 1H), 7.35 (d, J = 8.2 Hz, 1H), 7.13 (t, 1H), 7.06 (s, 1H), 7.00 (t, 1H), 3.71 (s, 3H), 2.24 (s, 3H).

b) 1,3-Dimethyl-1H-indole-2-carboxaldehyde

To a stirred solution of phosphorus oxychloride (7.0 mL, 75 mmol) in DMF (25 mL) was added dropwise a solution of 1,3-dimethylindole (12.0 g, 83 mmol) in dry DMF (6.0 mL). The reaction was stirred at room temperature for 2 h, then poured onto ice. The mixture was basified with a solution of NaOH (13.2 g, 330 mmol) in H 2 O (44 mL) then extracted with Et 2 O (2x 50 mL). The combined organic layers were washed with brine, dried (MgSO4), and concentrated in vacuo. Flash chromatography on silica gel (10% ethyl acetate / hexane) gave the title compound (13.03 g, 91%) as an off-white solid.

LCMS (ES) m / e 174.2 (M + H)⁺;

¹H NMR (400 MHz, CDCl3) δ 10.16 (s, 1H), 7.68 (d, J = 8.1 Hz, 1H), 7.42 (t, 1H), 7.32 (d, J = 8.5 Hz, 1H), 7.15 (t, 1H), 4.04 (s, 3H), 2.63 (s, 3H).

c) 1,3-Dimethyl-2- (methylaminomethyl) -1H-indole

To 1,3-dimethyl-1H-indole-2-carboxaldehyde (13.0 g, 75 mmol) was added a solution of 2.0 M methylamine in methanol (150 mL, 300 mmol) and HOAc (4.3 mL, 75 mmol) . The solution was stirred at room temperature for 4 h, then cooled to 0 ° C and sodium cyanoborohydride (5.0 g, 80 mmol) was added portionwise over 5 min. The reaction was then allowed to warm to room temperature. After 16 h, the reaction was concentrated under reduced pressure and the residue was dissolved in Et2O. The solution was washed with 1.0 N NaOH, then with brine, dried (Na2SO4), and concentrated to dryness. Flash chromatography on silica gel (95: 5 CHCl3 / methanol containing 5% NH4OH) gave the title compound (7.34 g, 52%) as a yellow oil.

¹H NMR (400 MHz, CDCl 3) δ 7.53 (d, J = 7.8 Hz, 1H), 7.26 (d, J = 7.8 Hz, 1H), 7.20 (t, 1H), 7.09 (t, 1H), 3.88 (s, 2H), 3.76 (s, 3H), 2.46 (s, 3H), 2.32 (s, 3H), 1.36 (br s, 1H).

Generation 41

Preparation of 6-bromo-2-oxo-1,4-dihydro-2H-pyrido [2,3-d] -1,3-oxazine

a) 2-Amino-3- (hydroxymethyl) pyridine

To a stirred solution of 2-aminonicotinic acid (20 g, 145 mmol) in dry THF (200 mL) under an argon atmosphere over 4 h, 1.0 M LiAlH4 in THF (300 mL, 300 mmol) was carefully added portionwise via reflux condenser. The reaction became exothermic and reflux with no external heat. After the addition was complete, the reaction was refluxed for an additional 16 h, then cooled to 0 ° C and carefully quenched by sequentially adding H 2 O (12 mL), 15% NaOH in H 2 O (12 mL). ) and H2O (35 mL). The resulting thick slurry was stirred for 1 h, then filtered through a pad of Celite^®. The filter pad was washed with THF (300 mL), and the filtrate was concentrated to dryness to obtain the title compound (17.04 g, 95%) as a pale yellow waxy solid.

LCMS (ES) m / e 125.1 (M + H)⁺;

¹H NMR (400 MHz, DMSO-d6) δ 7.84 (dd, 1H), 7.37 (m, 1H), 6.53 (dd, 1H), 5.65 (br s, 2H), 5, 16 (t, 1H), 4.34 (d, J = 4.6 Hz, 2H).

b) 2-Amino-5-bromo-3- (hydroxymethyl) pyridine

To a stirred solution of 2-amino-3- (hydroxymethyl) pyridine (15.0 g, 121 mmol) in HOAc (300 mL) at room temperature was added bromine (6.2 mL, 121 mmol) dropwise over 1 h. after approx. 15 min.

After the addition, the reaction was stirred for an additional 1 h then concentrated under reduced pressure. The residue was dissolved in 1.0 M Na2CO3 (500 mL) and the solution was extracted with ethyl acetate (2 x 250 mL). The combined organic layers were washed with brine, dried (Na2SO4) and concentrated to dryness. The resulting residue was triturated with a small volume of petroleum ether, filtered and dried under reduced pressure to provide the title compound (18.45 g, 75%) as a beige solid.

LCMS (ES) m / e 203.2 (M + H)⁺;

¹H NMR (400 MHz, DMSO-d6) δ 7.89 (d, J = 2.3 Hz, 1H), 7.52 (s, 1H), 5.92 (br s, 2H), 5.29 ( br s, 1H), 4.30 (s, 2H).

c) 6-Bromo-2-oxo-1,4-dihydro-2H-pyrido [2,3-d] -1,3-oxazine

To a stirred solution of 2-amino-5-bromo-3- (hydroxymethyl) pyridine (3.0 g, 15 mmol) in methanol (30 mL) was added dimethyl carbonate (5 mL, 60 mmol) and sodium methoxide (25 wt. methanol solution, 4 mL, 17.4 mmol).

The reaction was refluxed for 18 h, cooled to room temperature, and concentrated to dryness.

The residue was triturated with a saturated aqueous NH 4 Cl solution (50 mL), filtered, washed with cold H 2 O (50 mL) and dried under reduced pressure to provide the title compound (1.75 g, 51%) as a beige solid.

LCMS (ES) m / e 229.0 (M + H)⁺;

¹H NMR (400 MHz, DMSO-d6) δ 10.90 (s, 1H), 8.31 (s, 1H), 7.90 (s, 1H), 5.31 (s, 2H).

Generation 42

Preparation of 3-methyl-2- (methylaminomethyl) benzo [b] thiophene

To a stirred solution of 3-methylbenzo [b] thiophene-2-carboxaldehyde (0.5 g, 2.8 mmol) in methanol (15 mL) was added 2.0 M methylamine in methanol (6 mL, 12 mmol) and HOAc (0 , 32 mL, 5.7 mmol). The reaction was stirred at room temperature for an hour, then sodium cyanoborohydride (0.2 g, 3 mmol) was added in one portion.

After stirring for an additional 16 h, the reaction was concentrated to dryness. The residue was dissolved in and washed with 1.0 N NaOH, then brine, dried and concentrated in vacuo.

PL 201 627 B1

Purification by flash chromatography on silica gel (95: 5 CHCl3 / methanol containing 5% NH4OH) gave the title compound (0.30 g, 56%) as a yellow oil.

¹H NMR (400 MHz, CDCl 3) δ 7.77 (d, J = 7.8 Hz, 1H), 7.62 (d, J = 7.9 Hz, 1H), 7.34 (t, 1H), 7.28 (t, 1H), 3.99 (s, 2H), 2.49 (s, 3H), 2.34 (s, 3H), 1.77 (br s, 1H).

Generation 43

Preparation of 2- (methylaminomethyl) benzothiophene

a) N-methyl-benzothiophene-2-carboxamide

To a stirred solution of 2.0 M methylamine in THF (60 mL) and THF (60 mL) was added dropwise at 0 ° C a solution of benzothiophene-2-carbonyl chloride (10.8 g, 55 mmol) in THF (50 mL) in within 15 minutes.

After the addition, the reaction was allowed to warm to room temperature and then concentrated under reduced pressure.

Trituration with a cold 4: 1 H 2 O / methanol solution (50 mL), filtration and drying in vacuo gave the title compound (10.35 g, 98%) as a white solid.

MS (ES) m / e 191.9 (M + H)⁺.

b) 2- (Methylaminomethyl) benzothiophene

To a stirred suspension of N-methyl-benzothiophene-2-carboxamide (10.0 g, 52 mmol) in dry THF (75 mL) under argon was added a solution of 1.0 M LiAlH4 in THF (135 mL, 135 mmol) overnight. 15 minutes.

The reaction quickly became clear and was heated at reflux for 2 days. After cooling to 0 ° C, the reaction was carefully quenched by sequentially adding H 2 O (5.1 mL), 15% NaOH in H 2 O (5.1 mL), and H 2 O (15.3 mL).

_®

The mixture was filtered through a pad of Celite^® and the filter pad was washed with Et2O (50 mL). The filtrate was concentrated to provide the title compound (9.11 g, 99%) as a pale yellow oil which solidified in the freezer.

¹H NMR (400 MHz, CDCl3) δ 7.83 (d, J = 7.3 Hz, 1H), 7.72 (d, J = 7.3 Hz, 1H), 7.33 (m, 2H). 7.17 (s, 1H), 4.06 (s, 2H), 2.53 (s, 3H), 1.56 (br s, 1H).

Generation 44

Preparation of 2-methyl-3- (methylaminomethyl) indole

To a solution of 2-methylindole-3-carboxaldehyde (10.00 g, 62.84 mmol) in MeOH (100 mL) was added 2M CH3NH2 in MeOH (200 mL). After stirring for 3 hours at room temperature, the reaction solution was concentrated to a yellow oil which solidified under reduced pressure. This solid was dissolved in ethanol (350 mL) and NaBH4 (2.38 g, 62.8 mmol) was added.

The reaction was stirred at room temperature for 6 hours then concentrated under reduced pressure. The residue was diluted with saturated aqueous Na2CO3 solution (50 mL) and extracted with EtOAc (2 x 200 mL).

The organic phase was separated, washed with brine, and dried over Na2SO4. Flash chromatography on silica gel (9: 1 CHCl3 / MeOH containing 5% NH4OH) and drying in vacuo gave the title compound (6.88 g, 63%) as a pale yellow sticky solid.

MS (ES) m / e 175 (M + H)⁺.

Generation 45

Preparation of 5-bromo-2H-pyrido [3,2-b] -1,4-oxazin-3 (4H) -one

To a solution of 2H-pyrido [3,2-b] -1,4-oxazin-3 (4H) -one (5.00 g, 33.3 mmol) in HOAc (100 mL) was added Br2 (2.6 mL, 50.0 mmol). After stirring for 48 hours at room temperature, the reaction solution was concentrated to an orange solid which was suspended in 1N NaOH (50 mL) and extracted with EtOAc (2 x 100 mL). The combined organic layers were washed with brine and dried over Na2SO4. Flash chromatography on silica gel (9: 1 CHCl3 / MeOH containing 5% NH4OH) and drying in vacuo gave the title compound (5.49 g, 72%) as a yellow solid.

MS (ES) m / e 230 (M + H)⁺.

Generation 46

Preparation of 5-bromo-2-acetylaminopyrimidine

To a solution of 5-bromo-2-aminopyrimidine (2.0 g, 11.5 mmol) in CH2Cl2 (75 mL) at room temperature was added 2,6-lutidine (2.7 mL, 23.0 mmol) followed by the chloride acetyl (0.99 g, 12.6 mmol). After stirring for 3 hours, the reaction solution was concentrated under a reduced pressure.

PL 201 627 B1

The residue was dissolved in EtOAc (200 mL), washed with H 2 O (100 mL) and brine, and dried over Na 2 SO 4.

Flash chromatography on silica gel (95: 5 CHCl3 / MeOH) and drying in vacuo gave the title compound (1.74 g, 70%) as a yellow solid.

MS (ES) m / e 217 (M + H)⁺.

Production 47

Preparation of 1-Methyl-2- (methylaminomethyl) -6-methoxy-1H-indole

a) Methyl 1-methyl-6-methoxy-1H-indole-2-carboxylate

Following the procedure from Preparation 1 (a), except that methyl 6-methoxyindole-2-carboxylate was used in place of ethyl indole-2-carboxylate, the title compound (90%) was prepared as a beige solid.

MS (ES) m / e 220.2 (M + H)⁺.

b) N, 1-Dimethyl-6-methoxy-1H-indole-2-carboxamide

Following the procedure for Preparation 1 (b) except that methyl 1-methyl-6-methoxy-1H-indole-2-carboxylate was used in place of ethyl 1-methyl-1H-indole-2-carboxylate, the title compound ( 95%) as an off-white solid.

MS (ES) m / e 219.2 (M + H)⁺ and 437.4 (2M + H)⁺.

c) 1-Methyl-2- (methylamino methyl) -6-methoxy-1H-indole

Prepared following the procedure for preparing 1 (c) except that N, 1-dimethyl-6-methoxy-1H-indole-2-carboxamide was used in place of N, 1-dimethyl-1H-indole-2-carboxamide, the title (76%) as a light gray solid.

MS (ES) m / e 205.2 (M + H)⁺, 409.4 (2M + H)⁺.

Production 48

Preparation of 1,7-dimethyl-3- (methylaminomethyl) -1H-indole

a) 1,7-Dimethyl-1H-indole

Following the procedure in Preparation 1 (a), except that 7-methylindole was used in place of ethyl indole-2-carboxylate, the title compound (89%) was prepared as a beige solid.

MS (ES) m / e 146.2 (M + H)⁺.

b) 1,7-Dimethyl-1H-indole-3-carboxaldehyde

Following the procedure of Preparation 40 (b), except that 1,7-dimethyl-1H-indole was used in place of 1,3-dimethylindole, the title compound (82%) was prepared as a light beige solid.

MS (ES) m / e 174.2 (M + H)⁺.

c) 1,7-Dimethyl-3- (methylaminomethyl) -1H-indole

Following the procedure for Preparation 40 (c) except that 1,7-dimethyl-1H-indole-3-carboxaldehyde was used in place of 1,3-dimethyl-1H-indole-1-carboxaldehyde, the title compound (98% ) as a white, crystalline solid.

MS (ES) m / e 189.2 (M + H)⁺.

Generation 49

Preparation of 1,5-dimethyl-3- (methylaminomethyl) -1H-indole

a) 1,5-Dimethyl-1H-indole

According to preparation 1 (a), except that 5-methylindole was used in place of ethyl indole-2-carboxylate, the title compound (92%) was prepared as an amber oil.

MS (ES) m / e 146.2 (M + H)⁺.

b) 1,5-Dimethyl-1H-indole-3-carboxaldehyde

Following the procedure for Preparation 40 (b), except that 1,5-dimethyl-1H-indole was used in place of 1,3-dimethylindole, the title compound (82%) was prepared as a light beige solid.

MS (ES) m / e 174.2 (M + H)⁺.

c) 1,5-Dimethyl-3- (methylaminomethyl) -1H-indole

Following the procedure of Preparation 36, except that 1,5-dimethyl-1H-indole-3-carboxaldehyde was used in place of 1,3-dimethyl-1H-indole-1-carboxaldehyde, the title compound was prepared (89%) as an oil .

MS (ES) m / e 189.2 (M + H)⁺.

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Generation 50

Preparation of 1,6-dimethyl-3- (methylaminomethyl) -1H-indole

a) 1,6-Dimethyl-1H-indole

Following the procedure for Preparation 1 (a), except that 5-methylindole was used in place of ethyl indole-2-carboxylate, the title compound (96%) was prepared as an amber oil.

MS (ES) m / e 146.2 (M + H)⁺.

b) 1,6-Dimethyl-1H-indole-3-carboxaldehyde

Following the procedure in Preparation 40 (b), except that 1,5-dimethyl-1H-indole was used in place of 1,3-dimethylindole, the title compound (99%) was prepared as a light beige solid.

MS (ES) m / e 174.2 (M + H)⁺.

c) 1,6-Dimethyl-3- (methylaminomethyl) -1H-indole

Following the procedure of Preparation 36, except that 1,5-dimethyl-1H-indole-3-carboxaldehyde was used in place of 1,3-dimethyl-1H-indole-1-carboxaldehyde, the title compound was prepared (95%) as an oil .

MS (ES) m / e 169.2 (M + H)⁺.

Production 51

Preparation of 1-benzyl-3- (methylaminomethyl) -1H-indole

a) 3- (Methylaminomethyl) -1H-indole

To a solution of indole-3-carboxaldehyde (5.4 g, 34.1 mmol) in MeOH (30 mL) was added a solution of 2.0 M CH3NH2 in MeOH (51.3 mL, 102.6 mmol). The reaction was stirred at room temperature overnight then concentrated to a light yellow oil.

This oil was dissolved in EtOH (40 mL) and NaBH4 (1.3 g, 34.1 mmol) was added. After 16 h, the reaction was concentrated to a slurry and dissolved in 10% Na2CO3 (100 mL).

The aqueous solution was extracted with EtOAc (2 x 200 mL) and the combined organic fractions were dried over Na2SO4 and concentrated. Drying under high vacuum gave the title compound (5.2 g, 94%) as a pale yellow oil.

MS (ES) m / e 161 (M + H)⁺.

b) 3- [N- (Benzyloxycarbonyl) -N-methylaminomethyl] -1H-indole

N- (Benzyloxycarbonyloxy) succinimide (8.9 g, 35.7 mmol) was added to a solution of 3- (methylaminomethyl) -1H-indole (5.2 g, 32.5 mmol) and triethylamine (5.0 mL, 65.7 mmol) in DMF (100 mL) at room temperature.

The reaction was stirred overnight then concentrated under reduced pressure. The residue was diluted with water, and the mixture was extracted with ethyl acetate.

The combined extracts were dried over Na2SO4 and concentrated.

Flash chromatography on silica gel (33% ethyl acetate / hexane) gave the title compound (7.0 g, 74%) as an off-white solid.

MS (ES) m / e 295 (M + H)⁺.

c) 3- [N- (Benzyloxycarbonyl) -N-methylaminomethyl] -1-benzyl-1H-indole

NaH (60% dispersion in mineral oil, 0.15 g, 3.8 mmol) was added portionwise, allowing gas evolution, to the solution of 3- [N- (benzyloxycarbonyl) -N-methylaminomethyl] -1H-indole (0.7 g, 2.5 mmol) in DMF (25 mL) at 0 ° C.

When the NaH addition was complete, benzyl bromide (1.2 mL, 10.0 mmol) was added at 0 ° C. The reaction was stirred at 0 ° C for 15 minutes then at room temperature overnight.

The reaction was diluted with water and extracted with ethyl acetate. The combined extracts were dried over Na2SO4 and concentrated. Flash chromatography on silica gel (33% ethyl acetate / hexane) gave the title compound (0.9 g, 93%) as an off-white solid.

MS (ES) m / e 385 (M + H)⁺.

d) 1-Benzyl-3- (methylaminomethyl) -1H-indole

3- [N- (Benzyloxycarbonyl) -N-methylaminomethyl] -1-benzyl-1H-indole (0.9 g, 2.3 mmol) was added to the Pearlman's catalyst slurry (about 0.30 g in MeOH at room temperature in a Parr bottle The reaction was placed under 345 kPa (50 psi) H 2 and shaken for 5 h. The mixture was filtered through Celite.^® and the filter pad was washed with MeOH. The filtrate was concentrated to provide the title compound (0.5 g, 86%) as a light yellow solid.

MS (ES) m / e 251 (M + H)⁺.

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Generation 52

Preparation of 2-phenylamino-3-bromopyridine

A mixture of 2,5-dibromopyridine (10.2 g, 43 mmol) in aniline (25 mL) was stirred and refluxed for 3 h. The reaction was cooled to room temperature and most of the aniline was stripped under reduced pressure. The residue was dissolved in ethyl acetate and the solution was washed with 1.0 N Na2CO3, then brine, dried (Na2SO4) and concentrated in vacuo. Trituration with petroleum ether, filtration and drying in vacuo gave the title compound (7.20 g, 67%) as a beige solid.

¹H NMR (400 MHz, CDCl3) δ 8.25 (d, J = 2.4 Hz, 1H), 7.58 (dd, 1H), 7.31-7.39 (m, 4H), 7.11 (m, 1H), 6.79 (br s, 1H).

MS (ES) m / e 249.0 (M + H)⁺.

Generation 53

Preparation of 1,2-dimethyl-3- (methylaminomethyl) -1H-indole

a) 1,2-Dimethylindole-3-carboxaldehyde

A solution of POCl3 (7.0 mL, 75 mmol) in DMF (100 mL) was stirred for 5 minutes at 0 ° C, then 1,2-dimethylindole (10.0 g, 69 mmol) was added in one portion. The reaction was allowed to warm to room temperature and stirred for 4 h. The thick suspension was poured into ice water (300 mL) and the flask was washed with additional water (50 mL). The aqueous mixture was basified with a solution of NaOH (13.2 g, 330 mmol) in H 2 O (50 mL) and the thick suspension was filtered to collect the solid. It was washed with water and dried under reduced pressure to provide the title compound (11.59 g, 97%) as an off-white solid.

¹H NMR (400 MHz, CDCl 3) δ 10.07 (s, 1H), 8.09 (d, J = 7.9 Hz, 1H), 7.54 (d, J = 7.6 Hz, 1H), 7.21 (dt, 2H), 3.73 (s, 3H), 2.70 (s, 3H).

b) 1,2-Dimethyl-3- (methyliminomethyl) -1H-indole

A solution of 2M methylamine in methanol (100 mL, 200 mmol) was added to 1,2-dimethylindole-3-carboxaldehyde (11.50 g, 66.4 mmol). The reaction was stirred for 4 h at room temperature then concentrated to dryness to afford the crude title compound.

¹H NMR (400 MHz, CDCl3) δ 8.55 (d, J = 1.4 Hz, 1H), 8.16 (d, J = 7.5 Hz, 1H), 7.42 (d, J = 7 , 8 Hz, 1H), 7.15 (t, 1H), 7.07 (t, 1H), 3.68 (s, 3H), 3.41 (s, 3H), 2.55 (s, 3H ).

c) 1,2-Dimethyl-3- (methylaminomethyl) -1H-indole

1,2-Dimethyl-3- (methyliminomethyl) -1H-indole was dissolved in ethanol (200 mL) and NaBH4 (2.6 g, 68.7 mmol) was added portionwise at room temperature with stirring (vigorous gas evolution). After 16 h, the reaction was concentrated under reduced pressure and the residue was basified with an aqueous solution of 1.0 N NaOH (200 mL). The mixture was extracted with Et2O (250 mL) and the combined Et2O extracts were washed with brine, dried (Na2SO4) and concentrated. Purification by flash chromatography on silica gel (5-10% (5% NH4OH / MeOH) / CHCl3) gave the title compound (8.47 g, 68%) as an oil which solidified in the freezer.

¹H NMR (400 MHz, CDCl 3) δ 7.60 (d, J = 7.7 Hz, 1H), 7.29 (d, J = 8.0 Hz, 1H), 7.19 (t, 1H), 7.12 (t, 1H), 3.93 (s, 2H), 3.69 (s, 3H), 2.49 (s, 3H), 2.45 (s, 3H).

Generation 54

Preparation of 3- (methylaminomethyl) benzo [b] thiophene

To a stirred solution of 2M methylamine in methanol (75 mmol, 150 mmol) was added benzo [b] thiophene-3-carboxaldehyde (5.3 g, 33 mmol) and HOAc (4.3 mL, 75 mmol). The reaction was stirred at room temperature for 1 h, then NaBH3CN (2.1 g, 33 mmol) was added portionwise over 5 minutes. The reaction was stirred for an additional 16 h and then concentrated under reduced pressure. The residue was dissolved in Et2O (300 mL) and washed with 1.0 N NaOH (300 mL) then with brine, dried (Na2SO4) and concentrated. Purification by flash chromatography on silica gel (5% (5% NH4OH / MeOH) / CHCl3) gave the title compound (2.81 g, 48%) as a brown oil.

¹H NMR (400 MHz, CDCl3) δ 7.87 (2d, 2H), 7.40 (m, 2H), 7.32 (s, 1H), 4.02 (s, 2H), 2.56 (s , 3H), 1.5 (br s, 1H).

Generation 55

Preparation of 5-bromo-2,2'-dipyridylamine

Bromine (3.0 mL, 58.2 mmol) was added dropwise over 15 minutes to a stirred solution of 2,2'-dipyridylamine (10 g, 58.4 mmol) in HOAc (100 mL). The reaction quickly turned into a thick slurry. After 2 h, the reaction was concentrated under reduced pressure and the residue was purified by chromatography

Flash on silica gel (0.5% (5% NH4OH / MeOH) / CHCl3). The resulting residue was triturated with hexane and dried under reduced pressure to provide the title product (1.77 g, 12%) as an off-white solid.

¹H NMR (400 MHz, CDCl 3) δ 9.88 (s, 1H), 8.31 (s, 1H), 8.23 ​​(d, J = 4.8 Hz, 1H), 7.83 (m, 2H ), 7.67 (t, 1H), 7.62 (d, J = 8.4 Hz, 1H), 6.90 (t, 1H);

MS (ES) m / e 250.0 (M + H) ⁺.

The 5,5'-dibromo-2,2'-dipyridylamine (4.04 g, 21%) was also isolated as a white solid after trituration with hexane and drying in vacuo.

¹H NMR (400 MHz, CDCl3) δ 10.08 (s, 1H), 8.32 (d, J = 2.5 Hz, 2H), 7.88 (dd, 2H), 7.68 (d, J = 9.0 Hz, 2H);

MS (ES) m / e 328.0 (M + H)⁺.

Generation 56

Preparation of 2- (methylaminomethyl) -3-methylbenzo [b] thiophene

Following the procedures for the preparation of 53 (b) and (c) except that 3-methylbenzo [b] thiophene-2-carboxaldehyde (7.40 g, 42 mmol) was used in place of 1,2-dimethylindole-3-carboxaldehyde prepared the title compound (6.02 g, 75%) as a pale yellow oil which solidified in the freezer.

¹H NMR (400 MHz, CDCl 3) δ 7.77 (d, J = 7.8 Hz, 1H), 7.62 (d, J = 7.9 Hz, 1H), 7.34 (t, 1H). 7.28 (t, 1H), 3.99 (s, 2H), 2.49 (s, 3H), 2.34 (s, 3H), 1.77 (br s, 1H).

Generation 57

Preparation of 2-methyl-3- (methylaminomethyl) benzo [b] thiophene

a) 2-Methylbenzo [b] thiophene-3-carboxaldehyde

SnCl4 (20 mL, 67 mmol) was added over 5 min to a stirred solution of 2-methylbenzo [b] thiophene (5.0 g, 33.7 mmol) in CH2Cl2 (75 mL) at 0 ° C under an argon atmosphere. After 15 minutes, dichloromethylmethyl ether (3.7 mL, 41 mmol) was added. The reaction turned yellowish in color. The reaction was allowed to warm to room temperature and stirred for 16 h, then poured into ice water (200 mL). The aqueous mixture was acidified with 1N HCl (100 mL) and stirred until the suspension dissolved. The organic phase was separated, dried (MgSO4) and concentrated in vacuo. Purification by flash chromatography on silica gel (10% ethyl acetate / hexane) provided the title compound (5.83 g, 98%) as a white, crystalline solid.

¹H NMR (400 MHz, CDCl 3) δ 10.38 (s, 1H), 8.61 (d, J = 8.1 Hz, 1H), 7.77 (d, J = 8.0 Hz, 1H), 7.48 (t, 1H), 7.39 (t, 1H), 2.93 (s, 3H).

b) 2-Methyl-3- (methylaminomethyl) benzo [b] thiophene

According to the procedures for preparation 53 (b) and (c) except that 2-methyl-benzo [b] thiophene-3-carboxaldehyde (5.0 g, 28.4 mmol) was used in place of 1,2-dimethylindole -3-carboxaldehyde, the title compound (4.89 g, 90%) was prepared as an oil which solidified in the freezer.

¹H NMR (400 MHz, CDCl 3) δ 7.78 (d, J = 7.9 Hz, 1H), 7.75 (d, J = 7.9 Hz, 1H), 7.37 (t, 1H). 7.29 (t, 1H), 3.95 (s, 2H), 2.60 (s, 3H), 2.50 (s, 3H).

Generation 58

Preparation of 3,4-dimethyl-2- (methylaminomethyl) thieno [2,3-b] -thiophene

According to the procedure of Preparation 24 (a), except that 3,4-dimethylthien- [2,3-b] thiophene-2-carboxaldehyde (0.5 g, 2.5 mmol) was used instead of 1-methylindole. 2-carboxaldehyde, the title compound (0.28 g, 53%) was prepared as a colorless oil.

MS (ES) m / e 212 (M + H)⁺.

Generation 59

Preparation of 1-methyl-2- (methylaminomethyl) naphthalene

a) N, 1-Dimethylnaphthalene-2-carboxamide

Following the procedure of Preparation 20 (a) except that 1-methylnaphthalene-2-carboxylic acid (J. Org. Chem. 1965, 22, 3869; 0.3 g, 1.6 mmol) was used instead of acid 3 -methyl-2-inden-2-carboxylic acid, the title compound (0.3 g, 94%) was prepared as a white solid.

MS (ES) m / e 200 (M + H)⁺.

b) 1-Methyl-2- (methylaminomethyl) naphthalene

Following the procedure for Preparation 20 (b) except that N, 1-dimethylnaphthalene-2-carboxamide (0.3 g, 1.5 mmol) was used in place of N, 3-dimethylindene-2-carboxamide, the title compound was prepared (0.1 g, 36%) as a colorless oil.

MS (ES) m / e 186 (M + H)⁺.

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Generation 60

Preparation of 1-methyl-3- (methylaminomethyl) -1H-pyrrolo [2,3-b] pyridine

a) 1-Methyl-1H-pyrrolo [2,3-b] pyridine

Following the procedure for Preparation 40 (a) except that 7-azaindole (2.28 g, 1.83 mmol) was used in place of 3-methylindole, prepared the title compound (1.4 g, 58%) as a yellow oil .

MS (ES) m / e 133 (M + H)⁺.

b) 1-Methyl-1H-pyrrolo [2,3-b] pyridine-3-carboxaldehyde

Following the procedure from Preparation 40 (b) except that 1-methyl-1H-pyrrolo [2,3-b] pyridine (0.7 g, 5.3 mmol) was used in place of 1,3-dimethylindole, title compound (0.4 g, 47%) as a white solid.

MS (ES) m / e 161 (M + H)⁺.

c) 1-Methyl-3- (methylaminomethyl) -1H-pyrrolo [2,3-b] pyridine

According to the procedure for Preparation 40 (c) except that 1-methyl-1H-pyrrolo [2,3-b] pyridine-3-carboxaldehyde (0.4 g, 2.5 mmol) was used instead of 1.3 -dimethyl-1H-indole-2-carboxaldehyde gave the title compound (0.2 g, 45%) as yellow oil.

MS (ES) m / e 176 (M + H)⁺.

Generation 61

Preparation of 2,3-dihydro-8- (methylaminomethyl) -1H-3a-azacyclopenta [a] indene

a) 2,3-Dihydro-1H-3a-azacyclopenta [a] indene-8-carboxaldehyde

Following the procedure of Preparation 40 (b), except that 2,3-dihydro-1H-3a-azacyclopenta [a] indene was used (J. Med. Chem. 1965, 8, 700; 0.24 g, 1 , 53 mmol) instead of 1,3-dimethylindole, the title compound (0.17 g, 60%) was obtained as a yellow solid.

MS (ES) m / e 186 (M + H)⁺.

b) 2,3-Dihydro-8- (methylaminomethyl) -1H-3a-azacyclopenta [a] indene

According to the procedure of Preparation 40 (c) except that 2,3-dihydro-1H-3a-azacyclopenta [a] indene-8-carboxaldehyde (0.17 g, 0.92 mmol) was used instead of 1.3 -dimethyl-1H-indole-2-carboxaldehyde gave the title compound (0.1 g, 54%) as yellow oil.

MS (ES) m / e 201 (M + H)⁺.

The following examples illustrate methods for preparing the biologically active compounds of the present invention from intermediates as described in the previous preparations.

Example 1

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (SB 422805)

EDC (0.70 g, 3.7 mmol) was added to the solution of (E) -3- (6-aminopyridin-3-yl) acrylic acid (0.61 g, 3.7 mmol), 1-methyl-2- (methylaminomethyl) -1H-indole (0.65 g, 3.7 mmol), HOBt.H2O (0.50 g, 3.7 mmol) and triethylamine (0.52 mL, 3.7 mmol) in DMF (30 mL) at room temperature. The reaction was stirred overnight and then concentrated under reduced pressure. The residue was diluted with 5% NaHCO3 and extracted with CH2Cl2. The combined organic extracts were washed with brine and dried over MgSO4. Flash chromatography on silica gel (3% MeOH / CH2Cl2) gave a colorless solid which was triturated with Et2O and dried. The title compound (1.0 g, 83%) was obtained as a white solid.

¹H NMR (300 MHz, CDCl3) δ 8.20 (br s, 1H), 7.45-7.70 (m, 3H), 7.00-7.30 (m, 3H), 6.69 (d , J = 15.4Hz, 1H), 6.30-6.50 (m, 2H), 4.89 (s, 2H), 4.67 (br s, 2H), 3.68 (s, 3H ). 3.01 (s. 3H);

MS (ES) m / e 321 (M + H)⁺.

Analysis for C19H20N4O 0.40 H2O:

Calculated: C 69.66; H 6.40; N 17.10;

Found: C 69.99; H 6.27; N 16.84.

Example 2

Preparation of (E) -3- (4-aminophenyl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (SB 425537)

EDC (218 mg, 1.14 mmol) was added to a solution of 4-aminocinnamic acid hydrochloride (220 mg, 1.10 mmol), 1-methyl-2- (methylaminomethyl) -1H-indole (0.20 g, 1.15 mmol), HOBt · H 2 O (154 mg, 1.14 mmol) and triethylamine (0.20 mL, 1.43 mmol) in DMF (20 mL) at room temperature. The reaction was stirred overnight and then concentrated under reduced pressure. The residue was diluted with 5% NaHCO3 and extracted with CH2Cl2. The combined organic extracts were washed with brine

(2 x 30 mL) and dried over MgSO4. Flash chromatography on silica gel (3% MeOH / CH2Cl2) gave the title compound (68 mg, 19%) as a yellow foam.

¹H NMR (360 MHz, DMSO-d6, 330K) δ 7.46 (d, J = 7.8 Hz, 1H), 7.42 (d, J = 15.3 Hz, 1H), 7.37 (d , J = 8.3 Hz, 1H), 7.32 (d, J = 8.5 Hz, 2H), 7.06-7.15 (m, 1H), 6.94-7.03 (m, 1H), 6.81 (d, J = 15.3 Hz, 1H), 6.58 (d, J = 8.5 Hz, 2H), 6.33 (s, 1H), 5.25 (br s , 2H), 4.85 (s, 2H), 3.70 (s, 3H), 3.02 (s, 3H);

MS (ES) m / e 320 (M + H)⁺.

Analysis for C20H21N3O 0.20 H2O:

Calculated: C 74.37; H 6.68; N 13.01;

Found: C 74.21; H 6.60; N 12.80.

Example 3

Preparation of (E) -N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (pyridin-3-yl) acrylamide (SB 432263)

EDC (0.22 g, 1.14 mmol) was added to the solution of trans-3- (3-pyridyl) acrylic acid (0.17 g, 1.14 mmol), 1-methyl-2- (methylaminomethyl) -1H- indole (0.20 g, 1.15 mmol) and HOBt.H2O (0.15 g, 1.11 mmol) in DMF (1.0 mL) at room temperature.

The reaction was stirred overnight and then concentrated under reduced pressure. The residue was diluted with 5% NaHCO3 and extracted with CH2Cl2.

The combined organic extracts were washed with brine and dried over MgSO4.

Flash chromatography on silica gel (3% MeOH / CH2Cl2) followed by preparative TLC (3% MeOH / CH2Cl2) gave the title compound (0.14 g, 40%) as a white solid.

Spectrum analysis ¹H NMR (360 MHz, CDCl3) showed an approximately 8: 1 mixture of amide rotamers; for the main rotamer: δ 8.79 (s, 1H), 8.59 (d, J = 3.9 Hz, 1H), 7.84 (d, J = 7.6 H z, 1H), 7.76 (d, J = 15.5 Hz, 1H), 7.59 (d, J = 7.8 Hz, 1H), 7.38-7.48 (m, 2H), 7.19-7.27 ( m, 1H), 7.08-7.17 (m, 1H), 6.98 (d, J = 15.5 Hz, 1H), 6.51 (s, 1H), 4.94 (s, 2H ), 3.73 (s, 3H), 3.09 (s, 3H);

MS (ES) m / e 306 (M + H)⁺.

Analysis for C19H19N3O 0.20 H2O:

Calculated: C 73.86; H 6.33; N 13.60;

Found: C 73.52; H 6.32; N 13.43.

Example 4

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indazol-3-ylmethyl) acrylamide (SB-493389)

a) (E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indazol-3-ylmethyl) acrylamide

EDC (230 mg, 1.2 mmol) was added to a solution of (E) -3- (6-aminopyridin-3-yl) acrylic acid (164 mg, 1.0 mmol), 1-methyl-3- (methylaminomethyl) - 1H-indazole (210 mg, 1.2 mmol), HOBt.H2O (162 mg, 1.2 mmol), and Et3N (0.28 mL, 2.0 mmol) in dry DMF (5 mL) at room temperature. After 18 h, the mixture was concentrated.

Flash chromatography on silica gel (5% - EtOH / EtOAc) gave the title compound (238 mg, 74%) as a white foam.

¹H NMR (400 MHz, CDCl3) δ 8.24 (m, 1H), 7.90 (m, 1H), 7.65 (m, 2H), 7.35 (m, 2H), 7.09 (m , 1H), 6.73 (m, 1H), 6.50 (m, 1H), 5.04 (s, 2H), 4.83 (bs, 2H), 4.04 (s, 3H), 3 . 10 (s. 3H);

MS (ES) m / e 322 (M + H)⁺.

Example 5

Preparation of (E) -3- (3,4-dihydro-2H-pyrido [3,2-b] -1,4-oxazin-7-yl) -N-methyl-N- (1-methyl-1H-indole -2-ylmethyl) acrylamide (SB-509663)

a) (E) -3- (3,4-Dihydro-2H-pyrido [3,2-b] [1,4] oxazin-7-yl) -N-methyl-N- (1-methyl-1H -indol-2-ylmethyl) acrylamide

EDC (230 mg, 1.2 mmol) was added to the solution of (E) -3- (3,4-dihydro-2H-pyrido [3,2-b] -1,4-oxazin-7-yl) acrylic acid ( 206 mg, 1.0 mmol), 1-methyl-2- (methylaminomethyl) -1H-indole (209 mg,

1.2 mmol), HOBt.H2O (162 mg, 1.2 mmol), and Et3N (0.21 mL, 1.5 mmol) in dry DMF (5 mL) at room temperature. After 18 h, the mixture was concentrated.

Flash chromatography on silica gel (5% EtOH / EtOAc) gave the title compound (238 mg, 66%) as a yellow solid.

¹H NMR (400 MHz, d⁶-DMSO) δ 7.99-6.95 (m, 8H), 6.40 (s, 1H), 4.82 (s, 2H), 4.11 (bs, 2H), 3.72 (bs, 3H), 3.67 (bs, 2H), 3.08 (s, 3H); for the secondary rotamer δ 6.15 (s, 1H), 5.02 (s, 2H), 2.96 (s, 3H).

MS (ES) m / e 363 (M + H)⁺.

PL 201 627 B1

Example 6

Preparation of (E) -N-methyl-N - [(1-methyl-1H-indol-2-ylmethyl)] - 3- (5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl) ) acrylamide (SB-514613)

a) (E) -N-Methyl-N - [(1-methyl-1H-indol-2-ylmethyl)] - 3- (5,6,7,8-tetrahydro-1,8-naphthyridin-3- yl) -acrylamide

EDC (203 mg, 1.06 mmol) was added to the solution of (E) -3- (5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl) acrylic (180 mg, 0.88 mmol) ), 1-methyl-2- (methylaminomethyl) -1H-indole (185 mg, 1.06 mmol), HOBt.H2O (143 mg, 1.06 mmol) and Et3N (0.31 mL, 2.2 mmol) in dry DMF (5 mL) at room temperature. After 18 h, the mixture was concentrated. Flash chromatography on silica gel (10% EtOH / EtOAc) gave the title compound (222 mg, 70%) as a yellow solid.

¹H NMR (400 MHz, DMSO-d6) δ 7.99-6.82 (m, 8H), 6.40 (s, 1H), 4.82 (s, 2H), 3.67 (m, 2H) , 3.29 (m, 3H), 3.07 (m, 3H), 2.73 (m, 2H), 1.77 (m, 2H); for the secondary rotamer δ 6.16 (s, 1H), 5.00 (s, 2H).

MS (ES) m / e 361 (M + H)⁺.

Example 7

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (thieno- [2,3-b] thiophen-2-ylmethyl) acrylamide (SB-519689)

a) (E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (thieno [2,3-b] thiophen-2-ylmethyl) acrylamide

EDC (230 mg, 1.2 mmol) was added to a solution of (E) -3- (6-aminopyridin-3-yl) acrylic acid (164 mg, 1.0 mmol), 2- (methylaminomethyl) thieno [2.3 -b] thiophene (220 mg, 1.2 mmol), HOBt.H2O (162 mg,

1.2 mmol) and Et3N (0.35 mL, 2.5 mmol) in dry DMF (5 mL) at room temperature. After 18 h, the mixture was concentrated. Flash chromatography on silica gel (5% EtOH / EtOAc) gave the title compound (138 mg, 42%) as a beige solid.

¹H NMR (400 MHz, d⁶-DMSO) δ 8.15 (d, J = 2.0 Hz, 1H), 7.84 (bs, 1H), 7.57 (d, J = 5.2 Hz, 1H), 7.43 (d , J = 15.2 Hz, 1H), 7.27 (m, 2H), 6.44 (m, 2H), 4.75 (s, 2H), 3.13 (s, 3H); for the secondary rotamer δ 5.00 (s, 2H), 2.95 (s, 3H).

MS (ES) m / e 330 (M + H)⁺.

Example 8

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (thieno- [3,2-b] thiophen-2-ylmethyl) acrylamide (SB-528779)

a) (E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (thieno [3,2-b] thiophen-2-ylmethyl) acrylamide

EDC (230 mg, 1.2 mmol) was added to a solution of (E) -3- (6-aminopyridin-3-yl) acrylic acid (164 mg, 1.0 mmol), 2- (methylaminomethyl) thieno [3.2 -b] thiophene (220 mg, 1.2 mmol), HOBt.H2O (162 mg,

1.2 mmol) and Et3N (0.35 mL, 2.5 mmol) in dry DMF (5 mL) at room temperature. After 18 h the mixture was diluted with H 2 O and extracted with EtOAc (3 x). The combined organic layers were dried (MgSO4) and concentrated. The solid was dissolved in 1: 1 MeOH / H20 and filtered. The filtrate was concentrated to approximately 1/3 volume. The precipitate was collected by filtration, washed with H 2 O and dried under reduced pressure to obtain the title compound (139 mg, 42%) as a light beige solid.

¹H NMR (400 MHz, d⁶-DMSO) δ 8.15 (d, J = 2.0 Hz, 1H), 7.83 (bd, 1H), 7.61 (d, J = 5.2 Hz, 1H), 7.40 (m , 3H), 6.45 (m, 2H), 4.75 (s, 2H), 3.13 (s, 3H); for the secondary rotamer δ 5.00 (s, 2H), 2.95 (s, 3H).

MS (ES) m / e 330 (M + H)⁺.

Example 9

Preparation of (E) -3- (3H-imidazo [4,5-b] pyridin-6-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (SB-530561)

a) (E) -3- (3H-midazo [4,5-b] pyridin-6-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide

EDC (230 mg, 1.2 mmol) was added to the solution of (E) -3- (3H-imidazo [4,5-b] pyridin-6-yl) -acrylic acid (189 mg, 1.0 mmol), 1 -methyl-2- (methylaminomethyl) -1H-indole (209 mg, 1.2 mmol), HOBt.H2O (162 mg, 1.2 mmol) and Et3N (0.28 mL, 2.0 mmol) in dry DMF (5 mL) at room temperature. After 18 h, the mixture was diluted with H2O. The title compound (193 mg, 56%) was collected as a white solid by filtration, washed with H 2 O and dried under reduced pressure.

¹H NMR (400 MHz, d⁶-DMSO) δ 8.72 (s, 1H), 8.50 (s, 2H), 7.68 (d, J = 15.4 Hz, 1H), 7.45 (m, 3H), 7.13 (m, 1H), 7.01 (m, 1H), 6.43 (s, 1H), 4.87 (s, 2H), 3.70 (s, 3H), 3.15 (s, 3H) ; for the secondary rotamer δ 8.68 (s, 1H), 8.47 (s, 2H), 6.19 (s, 1H), 5.10 (s, 2H), 3.74 (s, 3H), 3 . 01 (s. 3H).

MS (ES) m / e 346 (M + H)⁺.

PL 201 627 B1

Example 10

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (6-methyl-6H-thieno [2,3-b] pyrrol-5-ylmethyl) acrylamide (SB-559455)

a) (E) -3- (6-Aminopyridin-3-yl) -N-methyl-N- (6-methyl-6H-thieno [2,3-b] pyrrol-5-ylmethyl) acrylamide

EDC (132 mg, 0.69 mmol) was added to a solution of (E) -3- (6-aminopyridin-3-yl) acrylic acid (95 mg, 0.58 mmol), 6-methyl-5- (methylaminomethyl) - 6H-thieno [2,3-b] pyrrole (142 mg, 0.69 mmol), HOBt.H2O (93 mg, 0.69 mmol), and Et3N (0.16 mL, 1.16 mmol) in dry DMF ( 3 mL) at room temperature.

After 18 h, the mixture was diluted with H 2 O and extracted with EtOAc (3 x).

The combined organic layers were dried (MgSO4) and concentrated.

The residue was dissolved in MeOH and collected by filtration to provide the title compound (65 mg, 34%) as a yellow solid.

¹H NMR (400 MHz, d⁶-DMSO) δ 8.15 (s, 1H), 7.81 (d, J = 8.1 Hz, 1H), 7.43 (d, J = 15.2 Hz, 1H), 6.96 (m , 2H), 6.43 (m, 3H), 4.70 (s, 2H), 3.61 (s, 3H), 3.00 (s, 3H); for the secondary rotamer δ 4.87 (s, 2H), 2.90 (s, 3H);

MS (ES) m / e 327 (M + H)⁺.

Example 11

Preparation of (E) -3- (2-aminopyrimidin-5-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (SB-480682)

Following the procedure of Example 1 except that (E) -3- (2-aminopyrimidin-5-yl) acrylic acid (0.50 g, 3.0 mmol) was used instead of (E) -3- ( 6-aminopyridin-3-yl) acrylic, the title compound (0.86 g, 89%) was prepared as an off-white solid.

MS (ES) m / e 322 (M + H)⁺.

Example 12

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (benzo [b] thiophen-2-ylmethyl) -N-methylacrylamide (SB-485779)

Following the procedure in Example 1 except that 2- (methylaminomethyl) benzo [b] thiophene (0.47 g, 2.68 mmol) was used in place of 1-methyl-2- (methylaminomethyl) indole, the title compound ( 0.71 g, 91%) as an off-white solid.

MS (ES) m / e 324 (M + H)⁺.

Example 13

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -2-butenamide (SB-492441)

Following the procedure of Example 1 except that (E) -3- (6-aminopyridin-3-yl) -2-methylacrylic acid (0.40 g, 2.24 mmol) was used instead of (E) - 3- (6-aminopyridin-3-yl) acrylic, the title compound (0.65 g, 87%) was prepared as an off-white solid.

MS (ES) m / e 335 (M + H)⁺.

Example 14

Preparation of (E) -3- (6-amino-2-methylpyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -acrylamide (SB-495124)

Following the procedure of Example 1 except that (E) -3- (6-amino-2-methylpyridin-3-yl) acrylic acid (0.40 g, 2.24 mmol) was used in place of the acid (E) -3- (6-aminopyridin-3-yl) acrylic, the title compound (0.70 g, 94%) was prepared as an off-white solid.

MS (ES) m / e 335 (M + H)⁺.

Example 15

Preparation of (E) -3- (6-Amino-5-methylpyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -acrylamide (SB-496351)

Following the procedure of Example 1 except that (E) -3- (6-amino-5-methylpyridin-3-yl) acrylic acid (1.00 g, 5.62 mmol) was used instead of acid (E) -3- (6-aminopyridin-3-yl) acrylic, the title compound (1.78 g, 95%) was obtained as an off-white solid.

MS (ES) m / e 335 (M + H)⁺.

PL 201 627 B1

Example 16

Preparation of (E) -3- [6-acetylamino) pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -acrylamide (SB-515905)

To a stirred suspension of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N - [(1-methyl-1H-indol-2-yl) methyl] acrylamide (0.50 g, 1.56 mmol) and NaHCO3 (0.51 g, 6.09 mmol) in THF (75 mL) were added acetic anhydride (0.38 g, 3.74 mmol). The reaction was refluxed for 24 h and then concentrated. The residue was extracted with EtOAc and purified over silica gel (95: 5 CHCl3 / CH3OH) to give the title compound (0.54 g, 96%) as an off-white solid.

MS (ES) m / e 363 (M + H)⁺.

Example 17

Preparation of (E) -3- (6-amino-5-methylpyridin-3-yl) -N- (benzo [b] thiophen-2-ylmethyl) -N-methylacrylamide (SB-515906)

Following the procedure of Example 1 except that (E) -3- (6-amino-5-methylpyridin-3-yl) acrylic acid (0.40 g, 2.24 mmol) was used in place of the acid (E) -3- (6-aminopyridin-3-yl) acrylic and 2- (methylaminomethyl) benzo [b] thiophene (0.44 g, 2.47 mmol) instead of 1-methyl-2- (methylaminomethyl) indole, the title compound was prepared (0.69 g, 91%) as an off-white solid.

MS (ES) m / e 338 (M + H)⁺.

Example 18

Preparation of (E) -3- (6-amino-5-methylpyridin-3-yl) -N-methyl-N- (naphthalen-2-ylmethyl) acrylamide (SB-515907)

Following the procedure of Example 1 except that (E) -3- (6-amino-5-methylpyridin-3-yl) acrylic acid (0.40 g, 2.24 mmol) was used in place of the acid (E) -3- (6-aminopyridin-3-yl) -acrylic and 2- (methylaminomethyl) naphthalene (0.42 g, 2.47 mmol) instead of 1-methyl-2- (methylaminomethyl) indole, the title compound (0, 65 g, 87%) as an off-white solid.

MS (ES) m / e 332 (M + H)⁺.

Example 19

Preparation of (E) -3- [6-acetylamino-5-methylpyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (SB-520771)

Following the procedure in example 16 except that (E) -3- (6-amino-5-methylpyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2- ylmethyl) acrylamide (0.47 g, 1.4 mmol) instead of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N - [(1-methyl-1H-indol-2-yl) ) methyl] acrylamide, the title compound (0.49 g, 93%) was prepared as an off-white solid.

MS (ES) m / e 377 (M + H)⁺.

Example 20

Preparation of (E) -3- [6-amino-5- (hydroxymethyl) pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (SB-528917)

Following the procedure of Example 1 except that (E) -3- [6-amino-5- (hydroxymethyl) pyridin-3-yl] acrylic acid (0.40 g, 2.1 mmol) was used instead of (E) -3- (6-Aminopyridin-3-yl) acrylic, the title compound (0.56 g, 77%) was prepared as an off-white solid.

MS (ES) m / e 351 (M + H)⁺.

Example 21

Preparation of (E) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3- ylo) acrylamide (SB-548797)

a) N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide

To a solution of 1-methyl-2- (methylaminomethyl) indole (0.78 g, 4.5 mmol) from the preparation of 1 and triethylamine (1.4 mL, 10.0 mmol) in CH2Cl2 (50 mL) at 5 ° C acryloyl chloride (0.41 mL, 4.95 mmol) was added.

After 45 min, the reaction solution was poured into H 2 O and the layers were separated.

The organic phase was dried over Na2SO4 and concentrated to provide the title compound as a yellow oil.

It was used directly without further purification.

b) (E) -N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3 -yl) acrylamide

Following the procedure from Preparation 2 (a) except that N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (0.90 g, 3.96 mmol) was used in place of benzyl acrylate and 6-bromo-3,4-di48

Hydro-1H-1,8-naphthyridin-2-one (0.60 g, 2.64 mmol) instead of 2-amino-5-bromopyridine, the title compound (0.85 g, 86%) was prepared as whitish solid.

MS (ES) m / e 375 (M + H)⁺.

Example 22

Preparation of (E) -3- [6-amino-5 - [(2-hydroxyethylamino) carbonyl] pyridin-3-yl] -N- (1-methyl-1H-indol-2-ylmethyl) -N-methylacrylamide ( SB-553829)

Following the procedure in Example 1 except that (E) -3- [6-amino-5 - [(2-hydroxyethylamino) carbonyl] pyridin-3-yl] acrylic acid (1.35 g, 5 4 mmol) instead of (E) -3- (6-aminopyridin-3-yl) acrylic acid, the title compound (1.95 g, 89%) was obtained as an off-white solid.

MS (ES) m / e 408 (M + H)⁺.

Example 23

Preparation of (E) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (3-methyl-9-oxo-1,2,3,4-tetrahydropyridine [2,3 -d] pyrimidin-6-yl) acrylamide (SB-577883)

a) N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide

To a solution of 1-methyl-2- (methylaminomethyl) indole (0.96 g, 5.5 mmol) from Preparation 1 and triethylamine (1.54 mL, 11.0 mmol) in CH2Cl2 (50 mL) at 5 ° C acryloyl chloride (0.48 mL, 6.0 mmol) was added. After 45 min, the reaction solution was poured into H 2 O and the layers were separated. The organic phase was dried over Na2SO4 and concentrated to provide the title compound as a yellow oil. It was used directly without further purification.

b) (E) -N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (3-methyl-2-oxo-1,2,3,4-tetrahydropyrid- [2, 3-d] pyrimidin-6-yl) acrylamide

Following the procedure from Preparation 2 (a) except that N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (1.25 g, 5.5 mmol) was used in place of benzyl acrylate and 6-bromo-3-methyl-3,4-dihydro-1H-pyrido [2,3-d] pyrimidin-2-one (0.80 g, 3.3 mmol) in place of 2-amino-5-bromopyridine, prepared the title compound (0.62 g, 49%) as an off-white solid.

MS (ES) m / e 390 (M + H)⁺.

Example 24

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (4-methyl-4H-thieno [3,2-b] pyrrol-5-ylmethyl) acrylamide (SB-587738)

Following the procedure in Example 1 except that 4-methyl-5- (methylaminomethyl) -4H-thieno [3,2-b] pyrrole (0.60 g, 3.3 mmol) was used instead of 1-methyl- 2- (methylaminomethyl) indole gave the title compound (0.90 g, 92%) as an off-white solid.

MS (ES) m / e 327 (M + H)⁺.

Example 26

Preparation of (E) -3- [6-aminopyridin-3-yl] -N-methyl-N- (3-methyl-1H-inden-2-ylmethyl) acrylamide (SB-554444)

EDC (0.383 g, 2.0 mmol) was added to a solution of (E) -3- (6-aminopyridin-3-yl) acrylic acid (0.328 g, 2.0 mmol), 3-methyl-2- (methylaminomethyl) hydrochloride indene (0.420 g, 2.0 mmol), HOBt.H2O (0.306 g, 2.0 mmol), and triethylamine (0.57 mL, 4.0 mmol) in anhydrous DMF (18 mL) at room temperature. The reaction was stirred overnight and concentrated under reduced pressure. The residue was diluted with 5% NaHCO3 and extracted with CH2Cl2. The combined organic extracts were washed with brine and dried over MgSO4. Flash chromatography on silica gel (3% MeOH / CH2Cl2) gave the title compound (0.33 g, 52%) as a colorless solid.

MS (ES) m / e 320.2 (M + H)⁺.

Analysis for C20H21N3O 0.4 H2O:

Calculated: C 73.57, H 6.72, N 12.86;

Found: C 73.94, H 6.92, N 12.50.

Example 27

Preparation of (E) -3- [6-aminopyridin-3-yl] -N- (1H-inden-2-ylmethyl) -N-methylacrylamide (SB-559447)

Following the procedure in Example 26, except that 2- (methylaminomethyl) indene hydrochloride was used in place of 3-methyl-2- (methylaminomethyl) indene hydrochloride, the title compound (0.23 g, 38%) was obtained as an off-white solid.

MS (ES) m / e 306.2 (M + H)⁺.

Analysis for C19H19N3O 0.125 H2O:

PL 201 627 B1

Calculated: C 74.18, H 6.30, N 13.64;

Found: C 74.21, H 6.25, N 13.27.

Example 28

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (4-methoxy-1-methyl-1H-indol-2-ylmethyl) -N-methylacrylamide (SB-574396)

Following the procedure in Example 26 except that 4-methoxy-1-methyl-2- (methylaminomethyl) -1H-indole hydrochloride was used in place of 3-methyl-2- (methylaminomethyl) indene hydrochloride, title compound (0.115 g, 68%) was obtained as an off-white solid.

MS (ES) m / e 351.2 (M + H)⁺.

Analysis for C20H22N4O2:

Calculated: C 68.55, H 6.32, N 15.98;

Found: C 68.15, H 6.33, N 15.73.

Example 29

Preparation of (E) -3- [6- (acetylamino) pyridin-3-yl] -N-methyl-N- (3-methyl-1H-inden-2-ylmethyl) acrylamide (SB-576040)

To the solution of (E) -3- [6-aminopyridin-3-yl] -N-methyl-N- (3-methyl-1H-inden-2-ylmethyl) acrylamide (0.159 g, 0.5 mmol) from Example 26 in anhydrous THF (20 mL) NaHCO3 (0.126 g, 1.5 mmol) was added followed by acetic anhydride (0.153 g, 0.15 mmol). The mixture was refluxed for 40 h then concentrated under reduced pressure. The residue was partitioned between H 2 O and EtOAc and the organic layer was dried over MgSO 4, filtered and concentrated. The residue was triturated with diethyl ether to afford the title compound (0.135 g, 74.8%) as an off-white solid.

MS (ES) m / e 362.2 (M + H)⁺.

Analysis for C22H23N3O2 0.25 H2O:

Calculated: C 72.20, H 6.47, N 11.47;

Found: C 72.42, H 6.45, N 11.07.

Example 30

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (1,4-dimethyl-1H-indol-2-ylmethyl) -N-methylacrylamide (SB-577946)

Following the procedure in Example 26 except that 1,4-dimethyl-2- (methylaminomethyl) -1H-indole hydrochloride was used in place of 3-methyl-2- (methylaminomethyl) indene hydrochloride, title compound (0.088 g, 52, 7%) obtained as an off-white solid.

MS (ES) m / e 335.2 (M + H)⁺.

Analysis for C20H22N4O 0.125 H2O:

Calculated: C 71.35, H 6.66, N 16.64;

Found: C 71.23, H 6.65, N 16.67.

Example 31

Preparation of (E) -N-methyl-N- (3-methyl-1H-inden-2-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3 -yl) acrylamide (SB-587735)

a) N-Methyl-N- (3-methyl-1H-inden-2-ylmethyl) acrylamide

To a solution of 3-methyl-2- (methylaminomethyl) indene hydrochloride (0.132 g, 0.63 mmol) from Preparation 19 and triethylamine (0.19 g, 1.89 mmol) in CH2Cl2 (6 mL) at 0 ° C was added a solution of acryloyl chloride (0.06 mL, 0.7 mmol) in CH2Cl2 (2 mL).

The reaction was stirred at 0 ° C for 1 h, then poured into water. The layers were separated, and the organic layer was washed with brine, dried over MgSO4, and concentrated under reduced pressure to give the title compound (0.145 g, quantitative) as an oily solid.

MS (ES) m / e 228.2 (M + H)⁺.

b) (E) -N-Methyl-N- (3-methyl-1H-inden-2-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3 -yl) acrylamide

A mixture of 6-bromo-3,4-dihydro-1H-1,8-naphthyridin-2-one (0.096 g, 0.42 mmol) from the preparation of 15 and N-methyl-N- (3-methyl-1H-inden- 2-ylmethyl) acrylamide (0.141 g, 0.62 mmol) in propionitrile (10 mL) was treated with (i-Pr) 2NEt (0.15 mL, 0.08 mmol), palladium acetate (0.014 g, 0.062 mmol) and ( o-Tolyl) 3P (0.025 g, 0.08 mmol) and the resulting mixture was heated under mild reflux. After 18 h the reaction was cooled, filtered through celite^® and concentrated.

PL 201 627 B1

Flash chromatography on silica gel (2% MeOH / CH2Cl2) gave the title compound (0.06 g, 41%) as a glassy solid.

MS (ES) m / e 374.2 (M + H)⁺

Analysis for C.₂₃H.₂₃N₃ABOUT₂ 1.25 H.₂ABOUT:

Calculated: C 69.76, H 6.41, N 10.61;

Found: C 69.86, H 6.67, N 10.51.

Example 32

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (1-methyl-1H-indol-2-ylmethyl) -N-propylacrylamide (SB-497614)

Following the procedure of Example 1 except that 1-methyl-2- (propylaminomethyl) -1H-indole (0.2 g, 1 mmol) was used in place of 1-methyl-2- (methylaminomethyl) -1H-indole. the title compound (0.14 g, 40%) was obtained as a white solid.

MS (ES) m / e 349 (M + H)⁺.

Example 33

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (5-fluoro-1-methyl-1H-indol-2-ylmethyl) -N-methylacrylamide

Following the procedure of Example 1 except that 5-fluoro-2- (methylaminomethyl) -1H-indole (0.192 g, 1 mmol) was used in place of 1-methyl-2- (methylaminomethyl) -1H-indole, the compound was prepared title (0.1 g, 30%) as a white solid.

MS (ES) m / e 339 (M + H)⁺.

Example 34

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (naphthalen-1-ylmethyl) acrylamide (SB-497651)

Following the procedure of Example 1 except that N-methyl-1- (methylaminomethyl) naphthalene hydrochloride (0.2 g, 1 mmol) was used in place of 1-methyl-2- (methylaminomethyl) -1H-indole, the compound was prepared title (0.09 g, 28%) as a white solid.

MS (ES) m / e 318 (M + H)⁺.

Example 35

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (benzofuran-2-ylmethyl) -N-methylacrylamide (SB-506505)

Following the procedure of Example 1 except that 2- (methylaminomethyl) -benzofuran (0.17 g, 1.1 mmol) was used in place of 1-methyl-2- (methylaminomethyl) -1H-indole, the title compound ( 0.10 g, 30%) as a white solid.

MS (ES) m / e 308 (M + H)⁺.

Example 36

Preparation of (E) -N-methyl-3- [6- (methylamino) pyridin-3-yl] -N- (1-methyl-1H-indol-2-ylmethyl) -acrylamide (SB-512196)

Following the procedure of Example 1 except that (E) -3- [6- (methylamino) pyridin-3-yl] acrylic acid (0.15 g, 0.84 mmol) was used instead of (E) - 3- (6-aminopyridin-3-yl) acrylic, the title compound (0.1 g, 37%) was prepared as a white solid.

MS (ES) m / e 335 (M + H)⁺.

Example 37

Preparation of (E) -3- [6- (dimefylamino) pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (SB-513112)

Following the procedure of Example 1 except that (E) -3- [6- (dimethylamino) pyridin-3-yl] acrylic acid (0.20 g, 1.0 mmol) was used instead of (E) - 3- (6-aminopyridin-3-yl) acrylic, the title compound (0.22 g, 63%) was prepared as a white solid.

MS (ES) m / e 349 (M + H)⁺.

Example 38

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-cyclopropyl-N- (1-methyl-1H-indol-2-ylmethyl) -acrylamide (SB-516973)

Following the procedure of Example 1 except that 2- (cyclopropylamino) -1-methyl-1H-indole (0.22 g, 1.1 mmol) was used in place of 1-methyl-2- (methylaminomethyl) -1H- indole, the title compound (0.154 g, 53%) was prepared as a white solid.

MS (ES) m / e 347 (M + H)⁺.

PL 201 627 B1

Example 39

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (quinolin-3-ylmethyl) acrylamide (SB-518459)

Following the procedure of Example 1 except that 3- (methylaminomethyl) quinoline (0.172 g, 1 mmol) was used in place of 1-methyl-2- (methylaminomethyl) -1H-indole, prepared the title compound (0.100 g, 31 %) as a white solid.

MS (ES) m / e 319 (M + H)⁺.

Example 40

Preparation of (E) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (6-methylpyridin-3-yl) acrylamide (SB-519700)

Following the procedure of Example 1 except that (E) -3- (6-methylpyridin-3-yl) acrylic acid (0.18 g, 1.1 mmol) was used instead of (E) -3- ( 6-aminopyridin-3-yl) acrylic, the title compound (0.11 g, 31%) was prepared as a white solid.

MS (ES) m / e 320 (M + H)⁺.

Example 41

Preparation of (E) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- [6- (2-oxopropylamino) pyridin-3-yl] acrylamide (SB-526142)

To a solution of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (0.12 g, 0.32 mmol) from In example 1, NaH (14 mg, 60% oil dispersion, 0.35 mmol) and 1-bromo-2,2-dimethoxypropane (0.05 mL, 0.37 mmol) were added in DMF (1 mL). After 18 h at room temperature, the reaction was complete by TLC analysis. The solvent was removed under reduced pressure and the residue was purified by preparative reverse phase HPLC (YMC CombiPrep^® ODS-A, 10% to 90% CH3CN / H2O + 0.1% TFA) to give the title compound (11.6 mg) as a pale yellow oil.

¹H NMR (400 MHz, MeOH-d4, 2: 1 mixture of rotamers, secondary rotamer data in italics) δ 9.28 and 9.22 (s, 1H), 8.60 and 8.52 (s, 1H), 8 , 25 and 8.15 (d, 1H), 7.68 (d, J = 16Hz, 1H), 7.50 (m, 1H), 7.35 (m, 3H), 7.15 (m, 1H), 7.02 (m, 1H), 6.55 and 6.25 (s, 1H), 5.05 and 4.95 (s, 2H), 3.72 and 3.68 (s, 3H) , 3.50 and 3.48 (s, 3H), 3.35 (s, 2H), 3.15 and 3.10 (s, 3H).

MS (ES +) m / e 376.3 (M + H)⁺.

Unreacted (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (68 mg) was also recovered.

Example 42

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (1H-indol-2-ylmethyl) -N-methylacrylamide (SB-525065)

EDC (0.30 g, 1.58 mmol) was added to a solution of 3- (6-aminopyridin-3-yl) acrylic acid (0.26 g, 1.58 mmol), 2- (methylaminomethyl) -1H-indole ( 0.23 g, 1.43 mmol), HOBt.H2O (0.21 g, 1.58 mmol) and diisopropylethylamine (0.51 mL, 2.86 mmol) in DMF (20 mL) at room temperature. The reaction was stirred overnight and then concentrated under reduced pressure. The residue was diluted with water and extracted with ethyl acetate. The combined organic extracts were washed with brine and dried over Na2SO4. Flash chromatography on silica gel (10% MeOH / CHCl3) gave the title compound (0.30 g, 68%) as a light yellow solid.

MS (ES) m / e 307 (M + H)⁺.

Example 43

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (1-ethyl-1H-indol-2-ylmethyl) -N-methylacrylamide (SB-530562)

EDC (0.84 g, 4.38 mmol) was added to a solution of 3- (6-aminopyridin-3-yl) acrylic acid (0.72 g, 4.38 mmol), 1-ethyl-2- (methylaminomethyl) - 1H-indole (0.75 g, 3.98 mmol), HOBt.H2O (0.59 g, 4.38 mmol) and diisopropylethylamine (1.40 mL, 7.96 mmol) in DMF (30 mL) at room. The reaction was stirred overnight and then concentrated under reduced pressure. The residue was diluted with water and extracted with ethyl acetate. The combined organic extracts were washed with brine and dried over Na2SO4. Flash chromatography on silica gel (5% MeOH / CHCl3) gave the title compound (0.40 g, 30%) as a light beige solid.

MS (ES) m / e 335 (M + H)⁺.

PL 201 627 B1

Example 44

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-3-ylmethyl) acrylamide (SB-517360)

EDC (0.35 g, 1.89 mmol) was added to a solution of 3- (6-aminopyridin-3-yl) acrylic acid (0.31 g, 1.89 mmol), 1-methyl-3- (methylaminomethyl) - 1H-indole (0.30 g, 1.72 mmol), HOBt.H2O (0.24 g, 1.89 mmol) and diisopropylethylamine (0.60 mL, 3.44 mmol) in DMF (20 mL) at room. The reaction was stirred overnight and then concentrated under reduced pressure. The residue was diluted with water and extracted with ethyl acetate. The combined organic extracts were washed with brine and dried over Na2SO4. Flash chromatography on silica gel (5% MeOH / CHCl3) gave the title compound (0.30 g, 55%) as a light yellow solid.

MS (ES) m / e 321 (M + H)⁺.

Example 45

Preparation of (E) -3- [6 - ((E) -but-2-enoylamino) pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (SB -535178)

Crotonic anhydride (0.29 mL, 1.96 mmol) was added to the solution of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl ) acrylamide (0.16 g, 0.49 mmol) and sodium bicarbonate (0.20 g, 2.45 mmol) in THF (30 mL) at room temperature and the reaction was refluxed under a nitrogen atmosphere. After 48 h, the reaction was concentrated under reduced pressure and the residue was diluted with water and extracted with ethyl acetate. The combined extracts were dried over Na2SO4 and concentrated under reduced pressure to provide the title compound (0.10 g, 53%) as a beige solid.

MS (ES) m / e 389 (M + H)⁺.

Example 46

Preparation of (E) -3- [6- (1,3-dioxo-1,3-dihydroisoindol-2-yl) -pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol- 2-ylmethyl) acrylamide (SB-562326)

Phthalic anhydride (0.81 g, 5.48 mmol) was added to the solution of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl). ) acrylamide (0.44 g, 1.37 mmol) and sodium bicarbonate (0.58 g, 6.85 mmol) in THF (70 mL) at room temperature and the reaction was refluxed under a nitrogen atmosphere. After 48 h, the reaction was concentrated under reduced pressure and the residue was purified by flash chromatography on silica gel (ethyl acetate). The title compound (0.21 g, 33%) was obtained as a white solid.

MS (ES) m / e 451 (M + H)⁺.

Example 47

Preparation of (E) -3- [6 - [(2-carboxybenzoyl) amino] pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (SB-566299)

Phthalic anhydride (0.81 g, 5.48 mmol) was added to the solution of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl). ) acrylamide (0.44 g, 1.37 mmol) and sodium bicarbonate (0.58 g, 6.85 mmol) in THF (70 mL) at room temperature and the reaction was refluxed under a nitrogen atmosphere. After 48 h, the reaction was concentrated under reduced pressure and the residue was diluted with water and extracted with ethyl acetate. The combined extracts were dried over Na2SO4 and concentrated. Flash chromatography on silica gel (10% MeOH / CHCl3) gave the title compound (0.10 g, 16%) as a light yellow solid.

MS (ES) m / e 469 (M + H)⁺.

Example 48

Preparation of (E) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- [6- (propionylamino) pyridin-3-yl] acrylamide (SB-577886)

Propionic anhydride (0.90 mL, 7.04 mmol) was added to the solution of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) ) acrylamide (0.56 g, 1.76 mmol) and sodium bicarbonate (0.74 g, 8.8 mmol) in THF (40 mL) at room temperature and the reaction was refluxed under a nitrogen atmosphere. After 48 h, the reaction was concentrated under reduced pressure and the residue was diluted with water and extracted with ethyl acetate. The combined extracts were dried over Na2SO4 and concentrated. Flash silica gel chromatography (ethyl acetate) gave the title compound (0.35 g, 53%) as a white solid.

MS (ES) m / e 377 (M + H)⁺.

PL 201 627 B1

Example 49

Preparation of (E) -3- [6- (3-Ethylureido) pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -acrylamide (SB-571655)

Ethyl isocyanate (0.13 mL, 1.68 minol) was added to the (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) solution ) acrylamide (0.27 g, 0.84 mmol) and triethylamine (0.29 mL,

2.1 mmol) in DMF (30 mL) at room temperature. The reaction was stirred for 6 days then concentrated under reduced pressure and the residue was diluted with water and extracted with ethyl acetate. The combined organic extracts were washed with brine, dried over Na2SO4 and concentrated. Flash silica gel chromatography (ethyl acetate) gave the title compound (80 mg, 24%) as a light yellow solid.

MS (ES) m / e 392 (M + H)⁺.

Example 50

Preparation of (E) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- [6- (3-methylureido) pyridin-3-yl] acrylamide (SB-579625)

Methyl isocyanate (0.18 mL, 3.05 mmol) was added to the solution of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl ) acrylamide (0.20 g, 0.61 mmol) and triethylamine (0.17 mL, 1.22 mmol) in DMF (20 mL) at room temperature. The reaction was stirred for 5 days then concentrated under reduced pressure and the residue was diluted with water and extracted with ethyl acetate. The combined organic extracts were washed with brine, dried over Na2SO4 and concentrated. Flash chromatography on silica gel (ethyl acetate) gave the title compound (0.10 g, 43%) as an off-white solid.

MS (ES) m / e 378 (M + H)⁺.

Example 51

Preparation of (E) -3- [6- (acetylamino) pyridin-3-yl] -N-methyl-N- (1-methyl-1H-indol-3-ylmethyl) -acrylamide (SB-576064)

Acetic anhydride (0.12 mL, 1.24 mmol) was added to the solution of (E) -3- (6-aminopyridin-3-yl) -N- (1-methyl-1H-indol-3-ylmethyl) -N- methylacrylamide (0.10 g, 0.31 mmol) and sodium bicarbonate (0.13 g, 1.55 mmol) in THE (20 mL) at room temperature and the reaction was refluxed under a nitrogen atmosphere. After 48 h, the reaction was concentrated under reduced pressure and the residue was diluted with water and extracted with ethyl acetate. The combined extracts were dried over Na2SO4 and concentrated. Flash silica gel chromatography (ethyl acetate) gave the title compound (50 mg, 45%) as a white solid.

MS (ES) m / e 363 (M + H)⁺.

Example 52

Preparation of (E) -3- (6-aminopyridin-3-yl) -2-methyl-N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -acrylamide (SB-497772)

To a stirred solution of (E) -3- (6-aminopyridin-3-yl) -2-methylacrylic acid HCl salt (0.5 g, 2.3 mmol) in dry 1: 1 DMF / CH2Cl2 (30 mL) at 1-Methyl-2- (methylaminomethyl) indole (0.42 g, 2.4 mmol), HOBt.H2O (0.32 g, 2.4 mmol), Et3N (0.66 mL, 4.7 mmol) was added at room temperature. ) and EDC (0.46 g, 2.4 mmol). After stirring for 24 h, the reaction was concentrated to dryness. The residue was dissolved in ethyl acetate and the solution was washed with H 2 O, dried (Na 2 SO 4) and concentrated in vacuo. Flash chromatography on silica gel (4% methanol / CHCl3) followed by trituration with ethyl acetate / hexane gave the title compound (0.55 g, 75%) as an off-white solid.

LCMS (ES) m / e 335.2 (M + H)⁺;

¹H NMR (300 MHz, DMSO-d6) δ 7.96 (s, 1H), 7.52 (d, J = 7.8 Hz, 1H), 7.48 (dd, 1H), 7.43 (d , J = 7.8Hz, 1H), 7.14 (t, 1H), 7.03 (t, 1H), 6.46 (d, J = 8.7Hz, 1H), 6.43 (s , 1H), 6.40 (s, 1H), 6.17 (br s, 2H).

Example 53

Preparation of 2- (6-aminopyridin-3-ylmethyl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (SB-498785)

Following the procedure of Example 52 except that 2- (6-aminopyridin-3-ylmethyl) acrylic acid HCl salt (0.50 g, 2.3 mmol) was used in place of (E) -3- acid HCl salt ( 6-aminopyridin-3-yl) -2-methylacrylic, the title compound (0.55 g, 75%) was obtained as an off-white solid after purification by flash chromatography on silica gel (4% methanol / CHCl3).

PL 201 627 B1

LCMS (ES) m / e 335.2 (M + H)⁺;

¹H NMR (300 MHz, DMSO-d6) δ 7.75 (d, J = 2.0 Hz, 1H), 7.50 (d, J = 7.6 Hz, 1H), 7.38 (d, J = 8.1 Hz, 1H), 7.22 (dd, 1H), 7.12 (t, 1H), 7.01 (t, 1H), 6.40 (d, J = 8.4 Hz, 1H ), 6.17 (s, 1H), 5.83 (br s, 2H), 5.23 (s, 1H), 5.14 (s, 1H), 4.73 (s, 2H), 3. 57 (s, 3H), 3.36 (s, 2H), 2.82 (s, 3H).

Example 54

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (naphthalen-2-ylmethyl) acrylamide (SB-497773)

To a stirred solution of (E) -3- (6-aminopyridin-3-yl) acrylic acid (0.30 g, 1.8 mmol) in 1: 1 DMF / CH2Cl2 (25 mL) was added 2- (methylaminomethyl) naphthalene ( 0.34 g, 2 mmol), HOBt.H2O (0.27 g, 2 mmol), Et3N (0.28 mL, 2 mmol), and EDC (0.38 g, 2 mmol). After stirring at room temperature for 16 h, the reaction was concentrated under reduced pressure. The residue was dissolved in ethyl acetate and the solution was washed with H 2 O, dried (Na 2 SO 4) and concentrated to dryness. Purification by flash chromatography on silica gel (4% methanol / CHCl3), trituration with 1: 1 ethyl acetate / hexane, filtration and drying in vacuo gave the title compound (0.49 g, 81%) as an off-white solid.

LCMS (ES) m / e 318.0 (M + H)⁺.

Example 55

Preparation of (E) -3- (6-amino-4-methylpyridin-3-yl) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -acrylamide (SB-515908)

Et3N was added to a stirred solution of (E) -3- (6-amino-4-methylpyridin-3-yl) acrylic acid (0.70 g, 3.3 mmol) in 1: 1 DMF / CH2Cl2 (30 mL). (0.42 mL, 3 mmol), 1-methyl-2- (methylaminomethyl) indole (0.50 g, 2.9 mmol), HOBt.H2O (0.41 g, 3 mmol) and DCC (0.70 g, 3 mmol). After stirring at room temperature for 16 h, the reaction was concentrated under reduced pressure. The residue was dissolved in ethyl acetate and filtered. The filtrate was washed with 1.0 N Na2CO3, then brine, dried (Na2SO4) and concentrated in vacuo. Purification by flash chromatography on silica gel (4% MeOH / CHCl3) gave the title compound (0.74 g, 74%) as a pale yellow solid.

LCMS (ES) m / e 335.2 (M + H)⁺.

Example 56

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (1,3-dimethyl-1H-indol-2-ylmethyl) -N-methylacrylamide (SB-532576)

To a stirred solution of 1,3-dimethyl-2- (methylaminomethyl) -1H-indole (0.6 g, 3.2 mmol) in 1: 1 DMF / CH2Cl2 (25 mL) was added acid (E) -3- (6 -aminopyridin-3-yl) acrylic (0.50 g, 3 mmol), HOBt.H2O (0.43 g, 3.2 mmol), and DCC (0.66 g, 3.2 mmol). After stirring at room temperature for 16 h, the reaction was concentrated under reduced pressure. Purification by flash chromatography on silica gel (3% methanol / CHCl3) gave the title compound (0.83 g, 83%) as an off-white solid.

LCMS (ES) m / e 335.4 (M + H)⁺.

Example 57

Preparation of (E) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (2-oxo-1,4-dihydro-2H-pyrido [2,3-d] -1,3-oxazin-6-yl) -acrylamide (SB-559456)

a) N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide

To a stirred solution of 1-methyl-2- (methylaminomethyl) -1H-indole (1.0 g, 5.7 mmol) from Preparation 1 and Et3N (0.9 mL, 6.4 mmol) in CH2Cl2 (50 mL) in at 0 ° C, acryloyl chloride (0.51 mL, 6 mmol) was added dropwise over 5 minutes. The reaction was stirred at 0 ° C for 1 h, then poured out into ice water. The organic phase was separated, washed with brine, dried (MgSO4) and concentrated to dryness to give the title compound (1.19 g, 91%) as a yellow oil. It was used without further purification.

TLC (silica gel, 50% EtOAc / hexane) Rf = 0.31.

b) (E) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (2-oxo-1,4-dihydro-2H-pyrido [2,3-d] - 1,3-oxazin-6-yl) acrylamide

To a mixed solution of N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (1.19 g,

5.2 mmol) in propionitrile (50 mL) was added 6-bromo-2-oxo-1,4-dihydro-2H-pyrido [2,3-d] -1,3-oxazine (1.1 g, 4, 9 mmol), DIEA (1.75 mL, 10 mmol), palladium (II) acetate (112 mg, 0.5 mmol), and tri-o-tolylphosphine (304 mg, 1.0 mmol). The reaction was purged with argon and refluxed for 16 h, then cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in CHCl3 and the solution was filtered through a pad of silica gel (3% methanol / CHCl3). The filtrate was concentrated and the residue was triturated with ethyl acetate, collected by filtration and dried in vacuo to provide the title compound (1.02 g, 55%) as an off-white solid.

LCMS (ES) m / e 377.4 (M + H)⁺.

Example 58

Preparation of (E) -N- (1,3-dimethyl-1H-indol-2-ylmethyl) -N-methyl-3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridine -3-yl) acrylamide (SB-573466)

a) N- (1,3-Dimethyl-1H-indol-2-ylmethyl) -N-methylacrylamide

To a stirred solution of 1,3-dimethyl-2- (methylaminomethyl) indole (1.5 g, 8 mmol) from Preparation 40 and Et3N (1.12 mL, 8 mmol) in CH2Cl2 (75 mL) at 0 ° C was added acryloyl chloride (0.65 mL, 8 mmol) dropwise over 5 minutes.

The reaction was stirred at 0 ° C for 1 h, then poured into ice water. The organic phase was separated, washed with brine, dried (MgSO4) and concentrated to dryness to give the title compound (1.7 g, 90%) as a yellow oil. It was used without further purification.

TLC silica gel (50% EtOAc / hexane) R f = 0.41.

b) (E) -N- (1,3-Dimethyl-1H-indol-2-ylmethyl) -N-methyl-3- (7-oxo-5,6,7,8-tetrahydro [1,8] naphthyridines -3-yl) acrylamide

To a stirred solution of N- (1,3-dimethyl-1H-indol-2-ylmethyl) -N-methylacrylamide (1.7 g, 7 mmol) in propionitrile (50 mL) was added 6-bromo-3,4-dihydro 1H-1,8-naphthyridin-2-one (1.16 g, 5.1 mmol), DIEA (1.8 mL, 10.3 mmol), palladium (II) acetate (112 mg, 0.5 mmol) , and tri-o-tolylphosphine (304 mg, 1.0 mmol).

The reaction was purged with argon and refluxed for 16 h, then cooled to room temperature and concentrated under reduced pressure.

Purification by flash chromatography on silica gel (5% methanol / CHCl3), trituration with ethyl acetate, filtration and drying in vacuo gave the title compound (1.17 g, 59%) as an off-white solid.

LCMS (ES) m / e 389.2 (M + H)⁺.

Example 59

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (3-methylbenzo [b] thiophen-2-ylmethyl) -acrylamide (SB-573537)

To a stirred solution of 3-methyl-2- (methylaminomethyl) benzo [b] thiophene (0.30 g, 1.6 mmol) in 1: 1 DMF / CH2Cl2 was added HOBt.H2O (0.27 g, 2 mmol) and DCC (0.41 g, 2 mmol). The reaction was stirred for 16 h then concentrated under reduced pressure.

The residue was dissolved in CHCl3, washed with H2O, dried (Na2SO4) and concentrated. Purification by flash chromatography on silica gel (4% methanol / CHCl3) gave the title compound (0.39 g, 72%) as a pale yellow solid.

LCMS (ES) m / e 338.2 (M + H)⁺.

Example 60

Preparation of (E) -3- (2-aminopyrimidin-5-yl) -N- (benzo [b] thiophen-2-ylmethyl) -N-methylacrylamide (SB-601395)

Following the procedure of Example 1 except that (E) -3- (2-aminopyrimidin-5-yl) acrylic acid (1.49 g, 7.1 mmol) was used instead of (E) -3- ( 6-aminopyridin-3-yl) acrylic and 2- (methylaminomethyl) thieno [2,3-b] thiophene (1.38 g, 7.8 mmol) instead of 1-methyl-2- (methylaminomethyl) indole, the compound was prepared title (2.04 g, 89%) as yellow solid.

MS (ES) m / e 325 (M + H)⁺.

Example 61

Preparation of (E) -3- (2-aminopyrimidin-5-yl) -N-methyl-N- (1-methyl-1H-indol-3-ylmethyl) acrylamide (SB-609846)

Following the procedure of Example 31 except that 1-methyl-3- (methylaminomethyl) indole (1.96 g, 8.6 mmol) was used in place of 3-methyl-2- (methylaminomethyl) indene hydrochloride and 2-amino -5-bromopyrimidine (1.0 g, 5.75 mmol) in place of 6-bromo-3,4-dihydro-1H-1,8-naphthyridin-2-one, prepared the title compound (1.44 g, 78%) as a yellow solid.

MS (ES) m / e 322 (M + H)⁺.

PL 201 627 B1

Example 62

Preparation of (E) -N-methyl-N- (1-methyl-1H-indol-3-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3- ylo) acrylamide (SB-611113)

Following the procedure of Example 31 except that 1-methyl-3- (methylaminomethyl) indole (0.75 g, 3.3 mmol) was used in place of 3-methyl-2- (methylaminomethyl) indene hydrochloride, the title compound was prepared. (0.59 g, 72%) as a light yellow solid.

MS (ES) m / e 375 (M + H)⁺.

Example 63

Preparation of (E) -3- [2-aminopyrimidin-5-yl] -N-methyl-N- (3-methyl-1H-inden-2-ylmethyl) acrylamide (SB-612035)

Following the procedure of example 31 except that 2-amino-5-bromopyrimidine (0.32 g, 1.84 mmol) was used in place of 6-bromo-3,4-dihydro-1H-1,8-naphthyridine- 2-one, the title compound (0.47 g, 80%) was prepared as a light yellow solid.

MS (ES) m / e 321 (M + H)⁺.

Example 64

Preparation of (E) -3- [2- (acetylamino) pyrimidin-5-yl] -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (SB-624954)

Following the procedure of Example 31 except that 1-methyl-2- (methylaminomethyl) indole (1.45 g, 8.33 mmol) was used in place of 3-methyl-2- (methylaminomethyl) indene hydrochloride and 2-acetylamino -5-bromopyrimidine (1.20 g, 5.55 mmol) in place of 6-bromo-3,4-dihydro-1H-1,8-naphthyridin-2-one, prepared the title compound (2.38 g, 43%) as a yellow solid.

MS (ES) m / e 364 (M + H)⁺.

Example 65

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (2-methyl-1H-indol-3-ylmethyl) acrylamide (SB-624955)

Following the procedure of Example 1 except that 2-methyl-3- (methylaminomethyl) indole (0.45 g, 2.58 mmol) was used in place of 1-methyl-2- (methylaminomethyl) indole, prepared the title compound ( 0.68 g, 90%) as yellow solid.

MS (ES) m / e 321 (M + H)⁺.

Example 66

Preparation of (E) -3- (2-aminopyrimidin-5-yl) -N- (1,2-dimethyl-1H-indol-3-ylmethyl) -N-methylacrylamide (SB-627551)

Following the procedure of Example 31 except that 1,2-dimethyl-3- (methylaminomethyl) indole (1.62 g, 8.62 mmol) was used in place of 3-methyl-2- (methylaminomethyl) indene hydrochloride and 2 -amino-5-bromopyrimidine (1.00 g, 5.75 mmol) instead of 6-bromo-3,4-dihydro-1H-1,8-naphthyridin-2-one, the title compound was prepared (1.33 g, 69 %) as a yellow solid.

MS (ES) m / e 336 (M + H)⁺.

Example 67

Preparation of (E) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (3-oxo-3,4-dihydro-2H-pyrido [3,2-b] -1,4-oxazin-7-yl) acrylamide (SB-628749)

Following the procedure of Example 31 except that 1-methyl-2- (methylaminomethyl) indole (1.17 g, 6.75 mmol) was used in place of 3-methyl-2- (methylaminomethyl) indene hydrochloride and 5-bromo -2H-pyrido [3,2-b] -1,4-oxazin-3- (4H) -one (1.03 g, 4.50 mmol) instead of 6-bromo-3,4-dihydro-1H-1 , 8-naphthyridin-2-one, prepared the title compound (0.90 g, 53%) as a light yellow solid.

MS (ES) m / e 377 (M + H)⁺.

Example 68

Preparation of (E) -N-methyl-N- (2-methyl-1H-indol-3-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3 -yl) acrylamide (SB-641197)

Following the procedure of Example 31 except that 2-methyl-3- (methylaminomethyl) indole (1.40 g, 8.00 mmol) was used in place of 3-methyl-2- (methylaminomethyl) indene hydrochloride, the title compound was prepared. (1.30 g, 65%) as a light yellow solid.

MS (ES) m / e 376 (M + H)⁺.

PL 201 627 B1

Example 69

Preparation of (E) -N-methyl-N- (1-methyl-1H-indol-3-ylmethyl) -3- (3-oxo-3,4-dihydro-2H-pyrido [3,2-b] -1,4-oxazin-7-yl) -acrylamide (SB-642050)

Following the procedure of example 31 except that 1-methyl-3- (methylaminomethyl) indole (0.38 g, 2.20 mmol) was used in place of 3-methyl-2- (methylaminomethyl) indene hydrochloride and 5-bromo -2H-pyrido [3,2-b] -1,4-oxazin-3 (4H) -one (0.32 g, 1.40 mmol) instead of 6-bromo-3,4-dihydro-1H-1, 8-naphthyridin-2-one, the title compound (0.26 g, 50%) was prepared as a light yellow solid.

MS (ES) m / e 377 (M + H)⁺.

Example 70

Preparation of (E) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3- yl) propionamide (SB-642051)

For a solution of (E) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3 -yl) acrylamide (0.15 g, 0.40 mmol) in dioxane was added Pd (OH) 2 at room temperature. The flask was closed with a septum through which a balloon containing hydrogen (1 atm) was passed. The reaction was stirred at room temperature overnight and then filtered through a pad of Celite^®, washing with methanol. The filtrate was concentrated to provide the title compound (0.14 g, 94%) as a light yellow solid.

MS (ES) m / e 378 (M + H)⁺.

Example 71

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (6-methoxy-1-methyl-1H-indol-2-ylmethyl) -N-methylacrylamide (SB-591549)

Following the procedure of example 1 except that 1-methyl-2- (methylaminomethyl) -6-methoxy-1H-indole was used instead of 1-methyl-2- (methylaminomethyl) -1H-indole, the title compound (50 %) as a light yellow solid.

MS (ES) m / e 351.4 (M + H)⁺.

Analysis for C.₂₀H.₂₂N₄ABOUT₂ 1.25 H.₂ABOUT:

Calculated: C 63.64, H 6.66, N 14.84;

Found: C 63.51, H 6.21, N 14.71.

Example 72

Preparation of (E) -3- (7-aminopyridin-3-yl) -N- (1,7-dimethyl-1H-indol-3-ylmethyl) -N-methylacrylamide (SB-634934)

Following the procedure of Example 1 except that 1,7-dimethyl-3- (methylaminomethyl) -1H-indole was used instead of 1-methyl-2- (methylaminomethyl) -1H-indole, the title compound was prepared (50%) as a light yellow solid.

MS (ES) m / e 335.2 (M + H)⁺.

Analysis for C20H22N4O2 0.5 H2O:

Calculated: C 69.99, H 6.76, N 16.31;

Found: C 70.02, H 6.59, N 16.43.

Example 73

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (1,5-dimethyl-1H-indol-3-ylmethyl) -N-methylacrylamide (SB-640668)

Following the procedure of Example 1 except that 1,5-dimethyl-3- (methylaminomethyl) -1H-indole was used in place of 1-methyl-2- (methylaminomethyl) -1H-indole, the title compound was prepared (33%) as a light yellow solid.

MS (ES) m / e 335.2 (M + H)⁺.

Analysis for C20H22N4O2 H2O:

Calculated: C 68.16, H 6.86, N 15.89;

Found: C 68.37, H 6.70, N 15.62.

Example 74

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (1,6-dimethyl-1H-indol-3-ylmethyl) -N-methylacrylamide (SB-641525)

Following the procedure of Example 1 except that 1,6-dimethyl-3- (methylaminomethyl) -1H-indole was used in place of 1-methyl-2- (methylaminomethyl) -1H-indole, the title compound was prepared (33%) as a light beige solid.

PL 201 627 B1

MS (ES) m / e 335.2 (M + H) +.

Analysis for C.₂₀H.₂₂N₄ABOUT₂ 0.375 H.₂ABOUT:

Calculated: C 70.41, H 6.64, N 16.42;

Found: C 70.40, H 6.61, N 16.19.

Example 75

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (1-benzyl-1H-indol-3-ylmethyl) -N-methylacrylamide (SB-642023)

EDC (0.42 g, 2.20 mmol) was added to a solution of 3- (6-aminopyridin-3-yl) acrylic acid (0.36 g, 2.20 mmol), 1-benzyl-3- (methylaminomethyl) - 1H-indole (0.50 g, 2.00 mmol), HOBt.H2O (0.30 g, 2.20 mmol) and diisopropylethylamine (0.70 mL, 4.00 mmol) in DMF (30 mL) at room. The reaction was stirred overnight and then concentrated under reduced pressure. The residue was diluted with water and extracted with ethyl acetate. The combined organic extracts were washed with brine and dried over Na2SO4. Flash chromatography on silica gel (10% MeOH / CHCl3) gave the title compound (0.48 g, 60%) as a light yellow solid.

MS (ES) m / e 397 (M + H)⁺.

Example 76

Preparation of (E) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- [6- (phenylamino) pyridin-3-yl] -acrylamide (SB-569303)

a) N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide

To a mixed solution of 1-methyl-2- (methylaminomethyl) -1H-indole (1.5 g, 8.6 mmol) and Et3N (1.35 mL, 9.6 mmol) in CH2Cl2 (75 mL) at 0 ° C acryloyl chloride (0.77 mL, 9.5 mmol) was added dropwise over 5 minutes. After 2 h, the reaction was washed with cold H 2 O, brine, dried (MgSO 4) and concentrated in vacuo. The residue was used without further purification.

b) (E) -N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- [6- (phenylamino) pyridin-3-yl] acrylic amide

N-Methyl-N- (1-methyl-1H-indol-2-ylmethyl) acrylamide (from Example 76 (a)) was dissolved in propionitrile (50 mL). To this solution was added 2-phenylamino-5-bromopyridine (1.3 g,

5.2 mmol), DIEA (1.8 mL, 10 mmol), Pd (OAc) 2 (112 mg, 0.5 mmol) and P (o-tol) 3 (304 mg, 1.0 mmol). The reaction was purged with argon, and then stirred at reflux for 16 h. After cooling to room temperature, the reaction was concentrated to dryness under reduced pressure. Flash chromatography on silica gel (5% methanol / CHCl3) followed by a second flash chromatography on silica gel (50-70% EtOAc / CHCl3) gave a residue which was triturated with EtOAc / petroleum ether. Filtration and drying in vacuo gave the title compound (1.42 g, 69%) as an off-white powder.

MS (ES) m / e 396.20 (M + H)⁺.

Example 77

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (1,2-dimethyl-1H-indol-3-ylmethyl) -N-methylacrylamide (SB-610285)

Acid (E) was added to a stirred solution of 1,2-dimethyl-3- (methylamino-methyl) -1H-indole (0.8 g, 4.2 mmol) in 1: 1 DMF / CH2Cl2 (30 mL) at room temperature -3- (6-aminopyridin-3-yl) acrylic (0.7 g, 4.3 mmol), Et3N (0.61 mL, 4.3 mmol), HOBt H2O (0.58 g, 4.3 mmol) and EDC (0.83 g,

4.3 mmol). After 16 h, the reaction was concentrated under reduced pressure and the residue was dissolved in EtOAc (100 mL). The solution was washed with 1.0 N Na2CO3 (100 mL), then brine, dried (Na2SO4), and concentrated. Purification by flash chromatography on silica gel (4% MeOH / CHCl3) followed by trituration with 1: 1 Et2O / petroleum ether and drying in vacuo gave the title compound (1.36 g, 97%) as an off-white solid.

MS (ES) m / e 335.2 (M + H)⁺.

Example 78

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (benzo [b] thiophen-3-ylmethyl) -N-methylacrylamide (SB-610300)

Following the procedure of Example 77 except that 3- (methylaminomethyl) benzo [b] thiophene (0.75 g, 4.2 mmol) was used in place of 1,2-dimethyl-3- (methylaminomethyl) -1H-indole gave the title compound (1.05 g, 83%) as an off-white solid.

MS (ES) m / e 324.2 (M + H)⁺.

PL 201 627 B1

Example 79

Preparation of (E) -N-methyl-N- (1-methyl-1H-indol-2-ylmethyl) -3- [6- (pyridin-2-ylamino) pyridin-3-yl] acrylamide (SB-618268)

Following the procedure of Example 76 (a) and (b) except that 5-bromo-2,2'-dipyridylamine (1.3 g, 5.2 mmol) was used in place of 2-phenylamino-5-bromopyridine. the title compound (1.54 g, 75%) was obtained as an off-white solid.

MS (ES) m / e 398.2 (M + H)⁺.

Example 80

Preparation of (E) -N- (1,2-dimethyl-1H-indol-3-ylmethyl) -N-methyl-3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridine -3-yl) acrylamide (SB-627696)

a) N-Methyl-N- (1,2-dimethyl-1H-indol-2-ylmethyl) acrylamide

Following the procedure of Example 76 (a) except that 1,2-dimethyl-3- (methylaminomethyl) -1H-indole (1.5 g, 8 mmol) was used in place of 1-methyl-2- (methylaminomethyl) -1H-indole, the title compound was prepared and used without further purification.

b) (E) -N- (1,2-Dimethyl-1H-indol-3-ylmethyl) -N-methyl-3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridine -3-yl) acrylamide

Following the procedure of Example 76 (b) except that 6-bromo-3,4-dihydro-1H-1,8-naphthyridin-2-one (1.3 g, 5.7 mmol) was used instead of 2 -phenylamino-5-bromopyridine gave the title compound (0.57 g, 26%) as a white solid.

MS (ES) m / e 389.19 (M + H)⁺.

Example 81

Preparation of (E) -N-methyl-N- (3-methylbenzo [b] thiophen-2-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3- ylo) acrylamide (SB-633857)

a) N-Methyl-N- (3-methylbenzo [b] thiophen-2-ylmethyl) acrylamide

Following the procedure of Example 76 (a) except that 2- (methylaminomethyl) -3-methylbenzo [b] thiophene (1.53 g, 8 mmol) was used in place of 1-methyl-2- (methylaminomethyl) -1H -indole, the title compound was prepared and used without further purification.

b) (E) -N-Methyl-N- (3-methylbenzo [b] thiophen-2-ylmethyl) -3- (7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3 -yl) acrylamide

Following the procedure of example 76 (b) except that 6-bromo-3,4-dihydro-1H-1,8-naphthyridin-2-one (1.3 g, 5.7 mmol) was used instead of 2 -phenylamino-5-bromopyridine gave the title compound (0.85 g, 33%) as an off-white solid.

MS (ES) m / e 392.2 (M + H)⁺.

Example 82

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl- (2-methylbenzo [b] thiophen-3-ylmethyl) acrylamide (SB-642016)

Compliance is not the procedure of Example 77, except using 2-methyl-3- (metylominometylo) benzo [b] thiophene (1.2 g, 6.1 mmole) for the 1,2-dimethyl-3- (methylaminomethyl ) -1H-indole, obtained the title compound (1.22 g, 59%) as a pale yellow solid.

MS (ES) m / e 338.2 (M + H)⁺.

Example 83

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (3,4-dimethylthieno [2,3-b] thiophen-2-ylmethyl) -N-methylacrylamide (SB-530622)

Following the procedure of Example 1 except that 3,4-dimethyl-2- (methylaminomethyl) thieno [2,3-b] thiophene (0.026 g, 0.126 mmol) was used in place of 1-methyl-2- (methylaminomethyl) -1H-indole, prepared the title compound (0.013 g, 72%) as a white solid.

MS (ES) m / e 358 (M + H)⁺.

Example 84

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl-H- (1-methylnaphthalen-2-ylmethyl) acrylamide (SB-609849)

Following the procedure of Example 1 except that 1-methyl-2- (methylaminomethyl) naphthalene (0.100 g, 0.54 mmol) was used in place of 1-methyl-2- (methylaminomethyl) -1H-indole, the title compound was prepared (0.088 g, 49%) as a white solid.

MS (ES) m / e 332 (M + H)⁺.

PL 201 627 B1

Example 85

Preparation of (E) -3- (6-aminopyridin-3-yl) -N-methyl-N- (1-methyl-1H-pyrrolo [2,3-b] pyridin-3-ylmethyl) acrylamide (SB-633875 )

Following the procedure of Example 1 except that 1-methyl-3- (methylaminomethyl) -1H-pyrrolo [2,3-b] pyridine (0.2 g, 1.14 mmol) was used instead of 1-methyl- 2- (methylaminomethyl) -1H-indole, the title compound (0.19 g, 52%) was prepared as a white solid.

MS (ES) m / e 322 (M + H)⁺.

Example 86

Preparation of (E) -3- (6-aminopyridin-3-yl) -N- (2,3-dihydro-1H-3a-azacyclopenta [a] inden-8-yl) -N-methylacrylamide (SB-641957)

Following the procedure of Example 1 except that 2,3-dihydro-8- (methylaminomethyl) -1H-3a-azacyclopenta [a] indene (0.100 g, 0.5 mmol) was used instead of 1-methyl- 2- (methylaminomethyl) -1H-indole gave the title compound (0.063 g, 36%) as a white solid.

MS (ES) m / e 347 (M + H)⁺.

Example 87

Composition of the parenteral dosage unit

A preparation which contains 20 mg of the compound of Example 1 as a sterile dry powder is prepared as follows: 20 mg of the compound is dissolved in 15 mL of distilled water.

The solution is sterilized by filtration into a multi-dose 25 mL ampoule and lyophilized.

The drug composition is reconstituted from the powder by adding 20 mL of 5% dextrose in water (D5W) for intravenous or intramuscular injection.

The dose is thus determined by the injection volume.

Serial dilution may be performed by adding a measured volume of this dosage unit to another volume of D5W for injection, or a measured dose may be added to another drug dispensing mechanism, such as an intravenous drip bottle or bag, or other injection-infusion system.

Example 88

The composition of the oral dosage unit

A capsule for oral administration is prepared by mixing and grinding 50 mg of the compound of Example 1 with 75 mg of lactose and 5 mg of magnesium stearate. The resulting powder is sieved and filled into a hard gelatin capsule.

Example 89

The composition of the oral dosage unit

A tablet for oral administration is prepared by mixing and granulating 20 mg of sucrose, 150 mg of calcium sulfate dihydrate and 50 mg of the compound of Example 1 with a 10% gelatin solution.

The wet granules are sieved, dried, mixed with 10 mg of starch, 5 mg of talc and 3 mg of stearic acid and compressed into a tablet.