Tricyclic tetrahydroquinoline antibacterial agents

ANTIBACTERIAL TRICYCLIC TETRA — HYDROKINOLINE AGENTS

Technical Domain of the Invention

The subject invention discloses novel tetrahydroquinoline compounds and related compounds and their pharmaceutical compositions, which exhibit useful antibacterial activity against a wide range of human and veterinary pathogens, including gram-positive and gram-negative aerobic bacteria, as well as anaerobic organisms.

Background Development of bacterial resistance to currently available antibacterial agents is a growing global health problem. Of particular concern are infections caused by multidrug-resistant pathogens. These bacteria are associated with significant morbidity and mortality. A number of possible solutions to the developing bacterial resistance problem have been suggested (Silver, LL; Bostian, KA Antimicrob. Chemother Agents. 1993, 37, 377). Above all, the best solution to the bacterial resistance dilemma remains identification and

ΡΕ1551849 development of structurally novel antibacterial agents using a single mechanism of action (Chopra, I. et al. JAMA, 1996, 275, 401).

For example, quinolones are a widely prescribed group of bacterial DNA gyrase inhibitors. DNA gyrase is a tetrameric enzyme composed of two GyrA subunits and two GyrB subunits that produce a negative DNA spiral through a sequence of strand breaks, DNA passage through the enzyme complex, and resealing. Quinolones act on the GyrA subunit. Its mechanism of action involves stabilization or entrapment of the cleaved gyrase-DNA complex. This inhibits the essential function of DNA gyrase and leads to cell death. It should also be noted that quinolones also inhibit, to varying degrees, bacterial topoisomerase IV, an essential tetrameric enzyme involved in initiating DNA decatenation, the process by which two daughter chromosomes are separated after division of a bacterial chromosome. Topo isomerase IV is composed of two ParC subunits and two ParE units that exhibit structural similarity to GyrA and GyrB, respectively. Representative quinolones include the fluoroquinolones ciprofloxacin, levofloxacin and gatifloxacin. Bacterial resistance to fluoroquinolones is becoming increasingly problematic (Kotilainen, P. et al. J. Infect. Dis. 1990, 161, 41-44. Trucksis, M. et al. Ann. Intern. Med. 1991 , 114, 424-426 Chen, DK et al N. Engl J. Med. 1999, 34, 233-239).

ΡΕ1551849

Bacterial DNA gyrase inhibitors that complement quinolone activity by inhibiting the GyrB subunit have also been identified. Coumarins, exemplified by novobiocin and cumermycin Al, and cyclothialidines are representative GyrB inhibitors that bind to the subunit ATP recognition point. Unfortunately, novobiocin has limited therapeutic value due to the observation of rapid development of resistance during treatment and other limitations (Kim,

<td>Ok et</td><td>there. Exp. Opin.</td><td>The R. Patents</td><td> 1998, 8, 959-969.</td>

<td>Maxwell,</td><td>A. Trends in</td><td>Microbiology,</td><td> 1997, 5, 102-109.</td>

<td>Maxwell,</td><td colspan="2">A. Mol. Microbiol. 1993,</td><td>9, 681-686). At</td>

<td colspan="2">cyclothialidines suffer</td><td>of the questions</td><td>of the metabolism of</td>

<td>drug</td><td>(Boehm, H.-J. et</td><td colspan="2">al. J. Med. Chem. 2000, 43, 2664-</td>

<td> 2674).</td><td>HC Richards,</td><td>Oxamnique:</td><td>A drug for the</td>

<td>Topics,</td><td>in The Role</td><td>of Organic</td><td>Chemistry in Drug</td>

Research, 1985, 271-289 discloses high schistosomicidal activity by several 2,3,4,4a, 5,6-hexahydro-1 H -pyrazino [1,2a] quinolines.

Nijhuis et al. , The Novel Two-Step Synthesis of Hexahydropyrazino [1,2-a] quinolines, Synthesis, Communications, vol. 7, 1987, 641-645 also discloses that several members of the 2,3,4,4a, 5,6-hexahydro-pyrazine [1,2a] quinoline heterocycle class exhibit activity against early developmental forms of schistonoma mansoni.

ΡΕ1551849

Summary of the Invention

In general, the invention illustrates structurally novel tetrahydroquinolines and related compounds or pharmaceutically acceptable salts thereof, methods of their production and their use as antibacterial agents.

In one aspect, the invention illustrates a compound of formula I, including its enantiomeric, diastereomeric or tautomeric isomers, or any pharmaceutically acceptable salt thereof;

<img file="PT1551849E_D0001.tif" />

Where,

R¹ is (C (= O) R⁶;

R² is

C (= O) R⁷;

Each R³ is independently (a) H, (b) R¹²(c) Oxo, (d) C1-6 alkyl₇ which is eventually partially

ΡΕ1551849 unsaturated and is eventually substituted by one or more R¹¹(e) C 3-8 cycloalkyl which is optionally partially unsaturated and optionally substituted by one or more R¹¹, (f) aryl, optionally substituted by one or more R⁸, (g) heteroaryl optionally substituted by one or more R⁸or (h) halo;

Each R⁴ is independently (a) H, (b) halo, (c) OR¹², (d) OC (= O) NR⁹R¹⁰, (e) SR¹², (f) S (O)_mR¹³, (g) NR⁹R¹⁰, (h) NR⁹S (0) _mR¹³, (i) NR⁹C (= 0) OR¹³, (j) phenyl optionally substituted by one or more R⁸, (k) heteroaryl optionally substituted by one or more R⁸, (l) cyan, (n) CONR⁹R¹⁰, (o) CO₂R¹², (p) C (= O) R¹³,

ΡΕ1551849 (q) C (= NOR¹²) R¹³, (r) S (0) _mNR⁹R¹⁰, (s) NR⁹C (= 0) -R¹²(t) C 1-7 alkyl which is optionally partially unsaturated and optionally substituted by one or more (u) C 3-8 cycloalkyl which is optionally partially unsaturated and optionally substituted by one or more R¹¹, (v) N₃, (w) het¹ eventually replaced by one or more R⁸, or (x) C (0) O-C1-4 alkyl¹²;

Each R⁵ is regardless

<td>(The)</td><td>H,</td>

<td>(B)</td><td>C1-7 alkyl which is eventually</td>

<td>unsaturated and</td><td>is eventually replaced by</td>

partially one or more (c) C cycloalkyl₃_₈ which is eventually partially unsaturated and is eventually substituted by one or more R¹¹, (d) aryl optionally substituted by one or more R⁸, or (e) heteroaryl optionally substituted by one or more R⁸;

R⁶ and R⁷ together form -N (R¹⁷) -C (0) -N (R¹⁷) - or -N (R¹⁷) -C (S) -N (R)¹⁷)

ΡΕ1551849

<td>(The)</td><td>H,</td>

<td>(B)</td><td>halo,</td>

<td>(ç)</td><td>OR¹²,</td>

<td>(d)</td><td>OCF₃,</td>

<td>(and)</td><td>SR¹²,</td>

<td>(f)</td><td>S (0)_mR¹³,</td>

<td>(g)</td><td>NR⁹R¹⁰,</td>

<td>(H)</td><td>NR⁹S (0)_mR¹³,</td>

<td>(i)</td><td>NR⁹C (= 0) OR¹³,</td>

<td>(j)</td><td>phenyl optionally substituted by halo,</td>

cyano, C1-7 alkyl, or C1-7 alkoxy, in the alkyl portion of C1-7 alkyl, or C1-7 alkoxy is optionally substituted by one or more R¹¹;

<td>(k)</td><td>heteroaryl optionally substituted by</td>

<td>halo alkyl</td><td>C1-7, or C1-7 alkoxy,</td>

<td>(D</td><td>cyan,</td>

<td>(m)</td><td>nitro,</td>

<td>(n)</td><td>CONR⁹R¹⁰,</td>

<td> (0)</td><td>co₂r¹²,</td>

<td>(P)</td><td>C (= 0) R¹³,</td>

<td>(q)</td><td>C (= NO0R¹²) R¹³,</td>

<td>(r)</td><td>S (O)_mNR⁹R¹⁰,</td>

<td>(s)</td><td>NR⁹C (= 0) -R¹²,</td>

<td>(t)</td><td>C1-7 alkyl which is possibly partially</td>

<td>unsaturated and</td><td>is eventually replaced by one or more</td>

<td>R¹¹,</td><td></td>

<td>(u)</td><td>C 3-8 cycloalkyl which is eventually</td>

Parcialmente1551849 partially unsaturated and is eventually substituted by one or more R¹¹, (v) -C (O) H, or (w) -het¹;

R⁹ and R¹⁰ are independently (a) H, (b) OR¹²(c) aryl optionally substituted by one or more R¹⁴(d) heteroaryl optionally substituted by one or more R¹⁴(e) C1-7 alkyl which is optionally substituted by one or more R¹¹, (f) C 3-8 cycloalkyl which is optionally substituted by one or more R¹¹, (g) (C = O) R¹³, or (h) R⁹ and R¹⁰ together with the nitrogen to which they are attached form morpholine, pyrrolidine, piperidine, thiazine and piperazine, each of morpholine, pyrrolidine, piperidine, thiazine and piperazine optionally being substituted by (a) oxo, (b) phenyl optionally substituted by one or more R¹⁴, (color¹², (d) SR¹²,

ΡΕ1551849

<td>(and)</td><td>nr¹²r¹²,</td><td></td>

<td>(f)</td><td>halo,</td><td></td>

<td>(g)</td><td>CO₂R¹²,</td><td></td>

<td>(H)</td><td>conr¹²r¹²,</td><td></td>

<td>(i)</td><td>C1-7 alkyl</td><td>which is eventually replaced</td>

<td>by oxo, halo,</td><td>OR¹², SR¹²,</td><td>C1-7 alkyl or substituents of</td>

<td>NR¹²R¹², or</td><td></td><td></td>

<td>(j)</td><td>cycloalkyl</td><td>C3-8 which is eventually</td>

<td colspan="2">partially unsaturated and</td><td>is eventually replaced by</td>

<td colspan="2">one or more oxo, halo, 0R^lz</td><td>', SR¹², C1-7 alkyl, or substituted</td>

<td>NR tuintes¹²</td><td>R¹²;</td><td></td>

<td>R¹² is</td><td></td><td></td>

<td>(The)</td><td>H,</td><td></td>

<td>(B)</td><td>C1-7 alkyl</td><td>which is eventually partially</td>

unsaturated and optionally substituted by oxo, halo, C1-7 alkyl or C1-7 alkoxy substituents, (c) C3-8 cycloalkyl which is optionally partially unsaturated and optionally substituted by one or more oxo, halo, C1-7 alkyl , or alkoxy substituents

(D) aryl optionally substituted by one or more halo, C1-7 alkyl, or substituents of C1-7 alkoxy, or (e) heteroaryl optionally substituted by one or more halo, C1-7 alkyl, or substituents of C1-7 alkoxy;

R¹³ is (a) C1-7 alkyl which is optionally or more often by oxo, halo, carboxyl, substituted alkoxy substituents C1-7 alkyl or

ΡΕ1551849 (b) cycloalkyl C₃_₈ which is optionally partially unsaturated and optionally substituted one or more times by oxo, halo, C1-7 alkyl or C1-7 alkoxy substituents, (c) aryl optionally substituted by one or more halo, C1-7 alkyl or alkoxy substituents (D) heteroaryl optionally substituted by a

<td>or more</td><td>halo,</td><td>C1-7 alkyl or substituents</td>

<td></td><td>(and)</td><td>-C (0) OH</td>

<td></td><td>R¹⁴</td><td>is</td>

<td></td><td>(The)</td><td>H,</td>

<td></td><td>(B)</td><td>halo,</td>

<td></td><td>(ç)</td><td>C1-7 alkyl,</td>

<td></td><td>(d)</td><td>OR¹²,</td>

<td></td><td>(and)</td><td>OCF₃,</td>

<td></td><td>(f)</td><td>SR¹²,</td>

<td></td><td>(g)</td><td>S (0)_mR¹³,</td>

<td></td><td>(H)</td><td>NR¹²R¹²,</td>

<td></td><td>(i)</td><td>NR¹²S (0)_mR¹³,</td>

<td></td><td>(d)</td><td>NR¹²C (= 0) OR¹³,</td>

<td></td><td>(k)</td><td>phenyl optionally substituted.</td>

<td>alkyl</td><td>C1-7,</td><td>or C1-7 alkoxy,</td>

<td></td><td> (1)</td><td>heteroaryl eventually</td>

<td colspan="2">halo alkyl</td><td>Ç₄7, or C alkoxy₄_7,</td>

<td></td><td>(m)</td><td>cyan,</td>

<td></td><td>(n)</td><td>nitro,</td>

<td></td><td> (0)</td><td>CONR¹²R¹²,</td>

<td></td><td>(P)</td><td>co₂r¹²,</td>

ΡΕ1551849 (q) C (= O) R¹³ (r) C (= NOR¹²) R¹³, (S) S (0) _mNR¹²R¹², (t) NR⁹C (= 0) -R¹², (u) C alkyl¹⁷ which is optionally partially and is optionally substituted by oxo, halo, C1-7 alkyl, or NR substituents¹²R¹²;

(v) C 3-8 cycloalkyl which is optionally parasaturated and optionally substituted by oxo,

SR¹²C 1-7 alkyl or NR substituents¹²R¹²;

unsaturated OR¹², SR¹², especially halo, OR¹²,

X is (a) - (C (R¹⁵) 2) n-, (b) - (C (R¹⁵) 2) m-0- (C (R¹⁵) 2) k-, (c) - (C (R¹⁵) 2) mS (0) m- (C (R¹⁵) 2) k-, or (d) - (C (R¹⁵) 2) m-NR¹⁶- (C (R¹⁵) 2) k-;

Each R¹⁵ is independently (a) H, (b) OR¹¹(c) Oxo, (d) C1-6 alkyl which is optionally substituted by one or more substituents of R¹¹, (e) cycloalkyl C₃_₈ which is eventually partially unsaturated and is optionally substituted one or more times by one or more substituents of R¹¹, (f) aryl optionally substituted by one or more R⁸, or (g) heteroaryl optionally substituted by one or more R⁸;

ΡΕ1551849

<td>(The)</td><td>H</td>

<td>(B)</td><td>OR¹²,</td>

<td>(ç)</td><td>(C = 0) R¹³,</td>

<td>(d)</td><td>(C = 0) OR¹³,</td>

<td>(and)</td><td>(C = 0) nr⁹r¹⁰,</td>

<td>(f)</td><td>s (0)_mR¹³,</td>

<td>(g)</td><td>S (0) _mNR⁹R¹⁰,</td>

<td>(H)</td><td>Ch-v alkyl which is optionally substituted</td>

by one or more substituents of R¹¹(i) C 3-8 cycloalkyl which is optionally partially unsaturated and optionally substituted by one or more substituents of R¹¹,

<td>(j)</td><td>aryl optionally substituted by one or</td>

<td>more R⁸, or</td><td></td>

<td>(k)</td><td>heteroaryl optionally substituted by a</td>

<td>or more R⁸;</td><td></td>

R¹⁷ is

<td>(The)</td><td>H,</td>

<td>(B)</td><td>-OH, and</td>

<td>(ç)</td><td>C1-4 alkyl;</td>

R²⁰ is

<td>(The)</td><td>H,</td>

<td>(B)</td><td>C1-7 alkyl which is possibly partially</td>

<td>unsaturated and</td><td>is eventually replaced by one or more</td>

<td>R¹¹,</td><td></td>

C1551849 (c) C 3-8 cycloalkyl which is optionally partially unsaturated and is optionally substituted by one or more R¹¹, (d) aryl optionally substituted by one or more R⁸, or (e) heteroaryl optionally substituted by one or more R⁸;

wherein aryl denotes a phenyl radical or an ortho-fused bicyclic carbocyclic radical having about nine to ten ring atoms in which at least one ring is aromatic;

where hereroaryl embraces a radical attached via a ring carbon or ring nitrogen to a monocyclic aromatic ring containing five or six ring atoms consisting of carbon and 1,2,3 or 4 heteroatoms, selected from oxygen (- 0-), sulfur (-S-), sulfur oxygenated as sulfinyl (S = 0) and sulfonyl (S (= 0)₂), or nitrogen N (Z) where Z is absent or is H, O, C1-4 alkyl, phenyl or benzyl, or a radical of an ortho-fused bicyclic heterocycle of about eight to ten

<td>atoms of</td><td>ring</td><td>derived from</td><td>same;</td><td></td><td></td>

<td>het¹</td><td>it is a</td><td>mono-ring or</td><td>bicyclic ring</td><td>five</td><td> (5) ,</td>

<td>six (6),</td><td>Seven</td><td>(7), or eight</td><td>(8) linked members</td><td>by C</td><td>or N,</td>

each mono-ring or bicyclic ring fully saturated or partially unsaturated and having 1-4

51551849 heteroatoms selected from the group consisting of oxygen, sulfur and nitrogen; being het¹ optionally substituted by 1-2 substituents selected from alkyl Ch-Ch, amino, alkylamino Ch-Ch, alkyloxy Ch-Ch, halogen, -OH, -CN, = O, = S;

each k is independently 0

1 or 2;

each m is independently 0 or 2;

each n is independently 1 or 3; and provided that:

the compound is not 2,3,4,4a-tetrahydro-1 ', 3' dimethyl spiro [1H-methylpyrazino [1,2-a] quinoline-5 (6H), 5 '(2') -pyrimidine] - 2'4'6 '(1Ή, 3'H) -trione.

In another aspect, the invention includes methods for synthesizing compounds of formula I. The method includes reacting an amine of formula III with a substituted aldehyde or ketone in an ortho-substituted aromatic ring with a halogen such as fluorine (and in certain cases chlorine) (formula II) of formula II in a polar, aprotic solvent, followed by methylenation with a compound of formula IV; and heating the reaction in a wide variety of solvents.

ΡΕ1551849

<img file="PT1551849E_D0002.tif" />

where X, R¹, R², R³, R⁴ and R⁵ are as defined above and Y¹ is H or alkyl.

Embodiments of the invention may include one or more of the following. R⁴ is independently (a) H, (b) alo, (e) SR¹², (f) S (O)_mR¹³, (g) NR⁹R¹⁰, (h) NR⁹S (0) _mR¹³, (i) NR⁹C (= 0) OR¹³, (j) phenyl optionally substituted by one or more R⁸, (k) heteroaryl optionally substituted by one or more R⁸, (l) cyan, (n) CONR⁹R¹⁰, (o) CO₂R¹², (p) C (= O) R¹³,

ΡΕ1551849

<td>(q)</td><td>C (= NO0R¹²</td><td>) R¹³,</td>

<td>(s)</td><td>NR⁹C (= 0)</td><td>-R¹²,</td>

<td>(t)</td><td>alkyl</td><td>C1-7 which is eventually partially</td>

unsaturated and is eventually substituted by one or more R¹¹, or (u) het¹ is eventually replaced by one or more R⁸.

For example, R⁴ is independently selected from H, Br, F, CF 3, CN, NH 2, -C (O) -OCH 3, -S-CH 3, S (O) 2 -CH 3, -N (OCH 3) -CH 3, -NH- C (O) -O-t-butyl, -NH-C (O) -CH 3, heteroaryl optionally substituted by one or more R⁸het¹ eventually replaced by one or more R⁸, -S (0) 2CH₃, or phenyl optionally substituted by one or more of NO₂, Cl, F, -OCH₃, and -OCF₃. R³ is Η. X is - (C (R¹⁵) 2) m-0 (C (R¹⁵)2)_k- or - (C (R¹⁵) 2) m-NR¹⁶- (C (R¹⁵) 2) _k-. X is -C (R¹⁵) 2-OC (R¹⁵) 2- or —C (R¹⁵) 2-NR¹⁶- (C (R¹⁵) 2 -. R¹⁵ is regardless

H, C1-7 alkyl optionally substituted by one or more substituents of R¹¹. X is -C (H) (C1-4 alkyl) -0-C (H) (C4-4 alkyl) - or -C (H) (¢ 4-4 alkyl) -NR¹⁶-C (H) (¢ 4-4 alkyl) -. 0 compound of the formula

<img file="PT1551849E_D0003.tif" />

<img file="PT1551849E_D0004.tif" />

independently, (b), (c), (d), (e), (f), or (g).

The compound of formula I may be, but is not limited to:

if

1,2,4,4a-Tetrahydro-2,4-dimethyl spiro [[1,4] oxazine [4,3a] quinoline-5 (6H)), 5 '(2'H) -pyrimidine] -2' , 4 ', 6' (1'H, 3'H) trione;

8-Bromo-1,2,4,4a-tetrahydro-2,4-dimethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H)), 5 '(2Ή) -pyrimidine ] 2 ', 4', 6 '(1Ή, 3 Ή) -trione;

8-Fluoro-1,2,4,4a-tetrahydro-2,4-dimethyl spiro [[1,4] oxazin [4,3-a] quinoline-5 (6H)), 5 '(2'H ) -pyrimidine] 2 ', 4', 6 '(1', 3 ') -trione;

1,2,4,4a-tetrahydro-2,4-dimethyl-8-trifluoromethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H)), 5 '(2'H) -pyrimidine] 2 ', 4', 6 '(1', 3 ') -trione;

1,1 ', 2,3', 4,4 ', 4a, 6'-Octrahydro-2,4', 6'-trioxospiro [[1,4] oxazine [4,3-a] quinoline-5 (67%)), 5 '(2'T) pyrimidine] -8carbonitrile;

1,2,4,4a-Tetrahydro-2,4-dimethyl-8carboxamide spiro [[1,4] oxazine [4,3-a] quinoline5 (6H)), 5 '(2'H) -pyrimidine] - 2 ', 4', 6 '(1', 3 ') -trione;

ΡΕ1551849

1,2,4,4a-Tetrahydro-2,4-dimethyl spiro [[1,4] oxazine [4,3a] quinoline-5 (6H)), 5 '(2'H) -pyrimidine] -2' , 4 ', 6' (1'H, 3'H) trione;

8-Bromo-1,2,4,4a-tetrahydro-2,4-dimethyl spiro [[1,4] piperazine [4,3-a] quinoline-5 (6H)), 5 '(2'H) -pyrimidine] 2 ', 4', 6 '(1', 3 ') -trione;

8-Bromo-1,2,4,4a-tetrahydro-cis-2,4-dimethylpiro [[1,4] oxazine [4,3-a] quinoline-5 (6H)), 5 '(2'H) pyrimidine] -4'-thioxo-2 ', 6' (1 ', 3') -dione;

8-Bromo-1,2,4,4a-tetrahydro-cis-2,4-dimethylpiro [[1,4] oxazine [4,3-a] quinoline-5 (6H)), 5 '(2'H) pyrimidine] -2 ', 4', 6 '(1'methyl, 3'methyl) -trione;

N- [1,1 ', 2,3', 4,4 ', 4a, 6'-Octahydro-2,4-dimethyl-2', 4 ', 6'trioxospiro [[1,4] oxazine [ 4,3-a] quinolone-5 (6H), 5 '(2'H) pyrimidinyl] -8-yl] acetamide;

1,1 ', 2,3', 4,4 ', 4a, 6'-Octahydro-2,4-dimethyl-2', 4 ', 6'trioxospiro [[1,4] oxazine [4,3 -a] quinolone-5 (6%), 5 '(2'H) pyrimidine] -8-ylcarbamate of tert-butyl;

8-Amino-1,2,4,4a-tetrahydro-2,4-dimethylspiro [[1,4] oxazine [4,3-a] quinolone-5 (6H), 5 '(2'H) monohydrochloride pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione;

ΡΕ1551849

8-Acetyl-1,2,4,4a-tetrahydro-2,4-dimethylpiro [[1,4] oxazine [4,3-a] quinoline-5 (6%), 5 '(2'H ) -pyrimidine] 2 ', 4', 6 '(1Ή, 3'H) -trione;

8-Ethanone-O-Methyloxime-1-1,2,4,4a-tetrahydro-2,4-dimethylspiro [[1,4] oxazine [4,3-a] quinoline-5 (6%), 5 '(2'H) pyrimidine] -2', 4 ', 6' (1'H, 3 ') trione;

1,2,4,4a-Tetrahydro-2,4-dimethyl-8 (methylsulfonyl) spiro [[1,4] oxazine [4,3-a] quinoline5 (6 H), 5 '(2'H) -pyrimidine] -2 ', 4', 6 '(1', 1 ', 3') -trione;

1,2,4,4a-Tetrahydro-2,4-dimethyl-8- (methylsulfinyl) spiro [[1,4] oxazine [4,3-a] quinoline-5 (6%), 5 '( 2'H) -pyrimidine] 2 ', 4', 6 '(1Ή, 3'H) -trione;

1,2,4,4a-Tetrahydro-2,4-dimethyl-8- (methylthio) spiro [[1,4] oxazine [4,3-a] quinol-5 (62 '), 5' ( 2Ή) -pyrimidine] 2 ', 4', 6 '(1Ή, 3'H) -trione;

2,3,4,4a-Tetrahydro-1 ', 3,3'-trimethylspiro [1,2] pyrazino [1,2-a] quinoline-5 (6%), 5' (2'H) -pyrimidine ] 2 ', 4', 6 '(1Ή, 3 Ή) -trione;

2,3,4,4a-Tetrahydro-3-methylspyro [1,2-pyrazino [1,2a] quinoline-5 (6%), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3, H) trione;

ΡΕ1551849

1,1-Dimethylethyl-8-cyano-1,1'2,3 ', 4,4', 4a, 6'-octahydro2 ', 4', 6'-trioxospiro [3'-pyrazine [1, 2-a] quinoline 5 (6H), 5 '(2'H) -pyrididine] -3-carboxylate;

9- (4-Chlorophenyl) -1,2,4,4a-tetrahydro-2,4-dimethylspiro [[1,4] oxazine [4,3-a] quinoline-5 (6%), 5 '( 2 'H) pyrimidine] -2', 4 ', 6' (1 '/ 1, 3' / ') - trione;

1,2,4,4a-Tetrahydro-2,4-dimethyl-9- [4-trifluoromethoxy) phenyl] spiro [[1,4] oxazine [4,3-a] quinoline 5 (6H), 5 ' (2'H) pyrimidine] -2 ', 4', 6 '(1', 1 ', 3') -trione;

1,2,4,4a-Tetrahydro-9- (methoxyphenyl) -2,4-dimethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 (6%), 5 '(2 'H) pyrimidine] 2', 4 ', 6' (1 'H, 3'H) -trione;

9- (3-Chloro-4-fluorophenyl) -1,2,4,4a-tetrahydro-2,4-dimethylpiro [[1,4] oxazine [4,3-a] quinoline 5 (6 H), 5 '( 2 'H) pyrimidine] -2', 4 ', 6' (1 '/ 1, 3' / ') - trione;

1,2,4,4a-Tetrahydro-2,4-dimethyl-9- (3-nitrophenyl) spiro [[1,4] oxazine [4,3-a] quinoline5 (6 H), 5 '(2'H ) pyrimidine] -2 ', 4', 6 '(1' / 1 ', 3' / ') - trione;

1,1 ', 2,3', 4,4 ', 4a, 6'-Octahydro-2,4-dimethyl-2', 4 ', 6'trioxospiro [[1,4] oxazine [4,3 -a] quinoline5 (6%), 5 (2'H) pyrimidin] -9-yl] benzonitrile;

ΡΕ1551849

1,2,4,4a-Tetrahydro-2,4-dimethyl-9- [4- (methylsulfonyl) phenyl] spiro [[1,4] oxazine [4,3-a] quinoline5 (6H), 5 ' (2'H) pyrimidine] -2 ', 4', 6 '(1' / 1, 3 '/ 1) -trione;

1,2,4,4a-Tetrahydro-2,4-dimethyl-9- (4pyridinyl) spiro [[1,4] oxazine [4,3-a] quinoline5 (6H), 5 '(2'H ) pyrimidine] -2 ', 4', 6 '(' H, 3 'tf) -trione;

Methyl-1,1'-2,3 ', 4,4a, 6'-Octahydro-2,4-dimethyl-2', 4 ', 6'trioxospiro [[1,4] oxazine [4,3- a] quinoline 5 (6H), 5 '(2'H) pyrimidine] -9-carboxylate;

Methyl-1,1'-2,3 ', 4,4a, 6'-Octahydro-2,4-dimethyl-2', 4 ', 6'trioxospiro [[1,4] oxazine [4,3- a] quinoline 5 (6H), 5 '(2'H) pyrimidine] -8-carboxylate;

1,2,3,3 ', 4,4', 4a, 6'-Octahydro-2 ', 4', 'trioxospiro [1,2-pyrazine [1,2-a] quinoline monohydrochloride -5 (6%), 5 '(2') pyrimidine] -8-carbonitrile; and

Other compounds of the invention include:

(2R, 4S, 4aS) -8-acetyl-9,10-difluoro-2,4-dimethyl-1,2,4,4atetrahydro-2'H, 6H-spiro [1,4-oxazine [4, 3-a] quinoline-5,5'pyrimidine] -2 ', 4', 6 '(1H, 3'H) -trione;

(2R, 4S, 4aS) -2,4-dimethyl-8- [5- (trifluoromethyl) -1,2,4oxadiazol-3-yl] -1,2,4,4a-tetrahydro-2'H, 6H-spiro [1,4oxazino [4,3-a] quinoline-5,5'-pyrimidine] -2 ', 4', 6 '(1'H, 3'H) trione;

51551849 (2R, 4S, 4aS) -2,4-dimethyl-8- (3-methyl-1,2,4-oxadiazol-5-yl) 1,2,4,4a-tetrahydro-2'H, 6H-spiro [1,4-oxazine [4,3a] quinoline-5,5'-pyrimidine] -2 ', 4', 6 '(1H, 3'H) -trione;

(2S, 4R, 4aR) -8-acetyl-10-fluoro-2,4-dimethyl-1,2,4,4a-tetrahydro-2'H, 6H-spiro [1,4-oxazine [4,3- a] quinoline-5,5'pyrimidine] -2 ', 4', 6 '(1H, 3'H) -trione;

(2R, 4S, 4aS) -2,4-dimethyl-8- (5-methyl-1,2,4-oxadiazol-3-yl) 1,2,4a-tetrahydro-2'H, 6H -spiro [1,4-oxazine [4,3a] quinoline-5,5'-pyrimidine] -2 ', 4', 6 '(1H, 3'H) -trione;

4-azido-3-iodobenzyl (2R, 4S, 4aS) -2,4-dimethyl-2 ', 4', 6 'trioxo-1,1'2,3', 4,4 ', 4a, 6'- octahydro-2'H, 6H-spiro [1,4oxazino [4,3-a] quinoline-5,5'-pyrimidine] -8-carboxylate;

Other specific compounds of formula I include:

rei- (2R, 4S, 4aS) -8-Acetyl-9,10-difluoro-1,2,4,4a-tetrahydro-2,4-dimethylspiro [[1,4] -oxazine [4,3-a] quinoline 5 (6A), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'A, 3'A) -trione;

rei- (2R, 4S, 4aS) -1,2,4,4a-Tetrahydro-2,4-dimethyl-8- [5- (trifluoromethyl) -1,2,4-oxadiazol-3-yl] spiro [[1,4] oxazine [4,3-a] quinoline-5 (6A), 5 '(2' A) -pyrimidine] 2 ', 4', 6 '(1'H, 3'H) -trione ;

ΡΕ1551849 rei- (2R, 4S, 4aS) - 8- [5- (Difluoromethyl) -1,2,4-oxadiazol-3-yl]

1,2,4a-tetrahydro-2,4-dimethylpiro [[1,4] oxazino [4,3a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] -2' , 4 ', 6' (1'H, 3'H) trione;

rei- (2R, 4S, 4aS) -1,2,4,4a-Tetrahydro-2,4-dimethyl-8- (3-methyl-1,2,4-oxadiazol-5-yl) spiro [[1, 4] -oxazine [4,3a] quinoline-5 (6%), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) trione;

rei- (2R, 4S, 4aS) -8-Acetyl-10-fluoro-1,2,4,4a-tetrahydro2,4-dimethylpiro [[1,4] oxoxino [4,3-a] quinoline5 ( 6A), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1 'A, 3'A) -trione;

(2A, 4S, 4aS) -9,10-Difluoro-1,2,4,4a-tetrahydro-2,4-dimethylpiro [[1,4] oxoxino [4,3-a] quinoline-5 (6A) , 5 '(2'H) pyrimidine] -2', 4 ', 6' (1 'A, 3'A) -trione;

rei- (2R, 4S, 4aS) -1,2,4,4a-Tetrahydro-2,4-dimethyl-8- (1,2,4 oxadiazol-3-yl) spiro [[1,4] - oxazine [4,3-a] quinoline 5 (6A), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'A, 3'A) -trione;

rei- (2R, 4S, 4aS) -8- (5-Cyclopropyl-1,2,4,4-oxadiazol-3-yl) 1,2,4a-tetrahydro-2,4-dimethyl spiro [[1 , 4] -oxazine [4,3a] quinoline-5 (6%), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) trione;

rel-4- [3- [2R, 4S, 4aS) -1, 1 ', 2,3', 4,4 ', 4a, 6'-Octahydro-2,4-dimethyl-2', 4 ', 6 'trioxospiro - [[1,4] oxazine [4,3-a] quinolineΡΕ1551849

4 (6 '), 5' (2 '') -pyrimidin] -8-yl] -1,2,4-oxadiazol-5-yl] benzonitrile;

rei- (2R, 4S, 4aS) -1,2,4,4a-Tetrahydro-2,4-dimethyl-8 (5,2-methyl-1,2,4-oxadiazol-3-yl) spiro [[1 , 4] -oxazine [4,3a] quinoline-5 (6%), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) trione;

rei- (2R, 4S, 4aS) -1,2,4,4a-Tetrahydro-2,4-dimethyl-8- [5 (methylthio) -1,3,4-thiadiazol-2-yl] spiro [ [1,4] -oxazine [4,3a] quinoline-5 (6%), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) trione ;

and re (4-Azido-3-iodophenyl) methyl (2A, 4S, 4aS) 1,1 ', 2,3', 4,4 ', 4a, 6'-octahydro-2,4-dimethyl 2' , 4 ', 6' trioxospiro [[1,4] oxazine [4,3-a] quinoline5 (6 J), 5 '(2' J) -pyrimidine] -8-carboxylate.

The above embodiments encompass all stereoisomeric, e.g., enantiomeric, diastereomeric and tautomeric forms of the named compounds.

The compounds of Formula I include at least one chiral center. Formula I covers both racemic and enantiomerically enriched forms of the compound of this invention. Racemic mixing is useful in the same way.

51551849 and for the same purposes as the most active enantiomer; The difference is that it is necessary to use more racemic material to produce the same antibacterial effect. One skilled in the art will appreciate that some of the claimed compounds have multiple chiral centers present. In these cases diastereomers are possible. All of these diastereomers, in racemic and enantiomerically enriched forms, are also included within the scope of the compounds of Formula I.

In another aspect, the invention illustrates a pharmaceutical composition comprising one or more compounds of formula I. The composition may include an enantiomerically enriched form of the compound of formula I. For example, the composition may include at least 50% (more typically at least 80%). % or 90% or more) of one enantiomer of a compound of formula I relative to the other enantiomers of the compound.

Advantageously and surprisingly, the compounds of this invention inhibit bacterial DNA gyrase, an ATP-dependent type II topoisomerase, unlike other known DNA gyrase inhibitors such as quinolones, coumarins, and cyclotialidines.

The term alkyl refers to branched and straight chain substituents and, unless otherwise indicated, include 1 to 10 carbon atoms, preferably 1 to 8 carbon atoms. Examples of alkyl substituents

No. 1,551,849 include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, iso-pentyl, n-hexyl and the like. Unless otherwise indicated, each alkyl substituent is optionally substituted by one or three groups selected from halo, N (R¹⁶)₂where R¹⁶ is defined above, phenyl optionally substituted by one or more R⁸, keto, amino, alkyl, C 1 -C 6 thioalkyl, C 1 -C 6 alkoxy, hydroxy, carboxy, C 1 -C 6 alkoxycarbonyl, nitrile, cycloalkyl, and a 5 or 6 membered carboxylic ring or heterocyclic ring having 1 or 2 heteroatoms selected from nitrogen, substituted nitrogen, oxygen and sulfur. Substituted nitrogen means nitrogen bearing C1 - C6 alkyl or phenyl (CH₂)_Pwhere p is 1,2, or 3.

Description of the Invention

The subject invention discloses tricyclic tetrahydroquinoline topoisomerase inhibitors. The compounds of this invention inhibit bacterial DNA gyrase, a topoisomerase II. These compounds have useful activity against aerobic and anaerobic bacteria and are effective against a number of human and veterinary pathogens. Representative organisms include, but are not limited to, Staphylococcus aureus, Staphylococcus epidermis, Enterococcus faecalis, Enterococcus faecium, Streptococcus pneumoniae, Streptococcus pyogenes, Chlamydophila pneumoniae, Haemofilus influenzae, Moraxella catarrhali, Escherichia coli

Klebsiella pneumoniae, Pseudomonas aeruginosa, Clostridium

ΡΕ1551849 spp., Peptostreptococcus spp. and Bacteroides spp. One skilled in the art will appreciate that the described organisms are merely representative and that other bacteria are included in the activity spectrum of the claimed compounds.

The compounds of formula I may be in the form of pharmaceutically acceptable salts. The term pharmaceutically acceptable salts refers to salts prepared from pharmaceutically acceptable bases and non-toxic acids. Pharmaceutically acceptable non-toxic bases and acids include inorganic bases, inorganic acids, organic acids and inorganic bases. Salts derived from inorganic bases include aluminum, ammonium, calcium, ferric, ferrous, lithium, magnesium, potassium, sodium, zinc and the like. Salts derived from non-toxic, pharmaceutically acceptable organic bases include salts of primary, secondary and tertiary amines, including substituted amines, natural substituted amines, cyclic amines such as arginine, betaine, caffeine, choline, N, N-dibenzylethylenediamine. , diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine and the like. Salts derived from inorganic acids include salts of hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, mineral acids sulfonic acid phosphoric acid acid

ΡΕ1551849 phosphorus and the like. Salts derived from pharmaceutically acceptable non-toxic organic acids include salts of carboxylic acids, di-carboxylic acids and C1-6 alkyl tricarboxylic acids such as acetic acid, propionic acid, fumaric acid, succinic acid, tartaric acid, maleic acid, adipic acid and citric acid. Other salts may be derived from aryl and alkyl sulfonic acids such as toluenesulfonic acids and the like.

Pharmaceutically acceptable salts may be obtained using standard procedures well known in the art, for example, by reacting a sufficiently basic compound such as an amine with a suitable acid that produces a physiologically acceptable anion. Alkali metal (e.g. sodium, potassium or lithium) or alkaline earth metal (e.g. calcium) salts of carboxylic acids may also be made.

By the term effective amount of a compound as referred to herein is meant a non-toxic but sufficient amount of the compound (s) to produce the desired effect. As indicated below, the exact amount required will vary from subject to subject, depending on the species, age and general condition of the subject, the severity of the disease being treated, the compound (s) used ( s), the mode of administration and the like. Thus, it is not possible to specify an exact effective amount. However, an appropriate effective amount may be determined by a person of ordinary skill in the art using only experimentation of

Rotina1551849 routine. The therapeutically effective amount of the compound of formula I that is administered and the dosage regimen for treating a disease condition with the compound of formula I and / or compositions containing the compound of formula I depends on a variety of factors, including age, the subject's weight, gender and clinical condition, the severity of the disease, the route and frequency of administration, and the particular compound (s) used and may therefore vary widely. 0 Dosage of the compound of formula I administered to a mammal may be from about 0.001 to about 100 mg / kg body weight / day. In general, the compound of formula I is a component of a pharmaceutical composition. Pharmaceutical compositions contain well known carriers and excipients in addition to the compound of formula I. The pharmaceutical compositions may contain the compound of formula I in a range from about 1 to about 1000 mg, such as from about 50 to about 800 mg. Generally, the pharmaceutical composition includes from about 0.5% to about 90% by weight of the compound of formula I. A total daily dose of about 1 to 1000 mg of the compound of formula I may be appropriate for an adult. The daily dose may be given as one to four doses per day. The desired dose may conveniently be presented in a single dose or divided into multiple doses administered at appropriate intervals, for example as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, for example, into a number of freely spaced discrete administrations.

Also, it should be understood that the starting dose

The administered Δ1551849 can be increased beyond the above level to quickly achieve the desired plasma concentration. On the other hand, the starting dose may be less than optimal and the daily dose may be progressively increased over the course of treatment depending on the particular situation.

Pharmaceutical compositions of the compound of formula I, either individually or in combination with other antimicrobial agents, may be prepared by methods well known in the art, for example by conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying processes. encapsulation, entrapment, lyophilization or spray drying. Pharmaceutical compositions for use in accordance with the present invention may be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate the processing of the active compounds into preparations which may be used pharmaceutically. Proper formulation depends on the route of administration chosen.

The compounds of formula I may be administered parenterally, orally topically, transdermally and rectally (for example, as a suppository).

Formulations for systemic administration may be in the form of aqueous solutions and suspensions, in addition to tablet and capsule formulations. The solutions

Aqueous Δ1551849 and suspensions may be prepared from sterile powders or granules having one or more of the mentioned carriers or diluents for use in formulations for oral administration. The compounds may be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride and / or various buffers. Other adjuvants are well and widely known in the pharmaceutical art. For example, suspensions or solutions for systemic administration may include β cyclodextrins, such as Captisol®, as solubilizing agent (s). The compositions may, for example, be administered parenterally, for example intravascularly, intraperitoneally, subcutaneously or intramuscularly. For parenteral administration saline, dextrose solution or water may be used as a suitable carrier. Formulations for parenteral administration may be in the form of isotonic aqueous or non-aqueous solutions for sterile injection or suspensions.

Pharmaceutical compositions for parenteral administration will generally contain a pharmaceutically acceptable amount of the compound or a soluble salt (acid addition salt or base salt) dissolved in a pharmaceutically acceptable liquid carrier such as water for injection and a buffer to produce an isotonic solution. suitably buffered, for example having a pH of about 3.5-6. Suitable buffering agents include, for example, trisodium orthophosphate, bicarbonate

ΡΕ1551849 sodium, sodium citrate, N-methylglucamine, L (+) - lysine and L (+) - arginine to name but a few representative buffering agents. The compound of this invention will generally be dissolved in the carrier in an amount sufficient to produce a pharmaceutically acceptable injectable concentration in the range of from about 1 mg / mL to about 400 mg / mL of solution. The resulting liquid pharmaceutical composition will be administered so as to obtain the aforementioned amount of antibacterially effective dosage.

For systemic administration, the compounds may be formulated by combining the active compounds with pharmaceutically acceptable carriers well known in the art.

Such carriers allow the compounds of the invention to be formulated as tablets, pills, lozenges, dragées, capsules, liquids, solutions, emulsions, gels, syrups, slurries, suspensions and the like for oral ingestion by the patient.

In addition to the compound of formula I, the pharmaceutical composition for therapeutic use may also comprise one or more non-toxic, pharmaceutically acceptable carrier materials or excipients. The term carrier material or excipient herein means any substance other than a therapeutic agent used as a support and / or diluent and / or adjuvant, or vehicle for delivery of a therapeutic agent used as a support and / or diluent and / or adjuvant, or vehicle for delivering a therapeutic agent to a subject or added to a composition

51551849 to improve its handling or storage properties or to enable or facilitate the formation of a composition dosage unit in a discrete article such as a capsule or tablet suitable for oral administration. Excipients may include, by way of illustration and not limitation, diluents, disintegrants, binding agents, adhesives, wetting agents, polymers, lubricants, glidants, added substances to mask or combat an unpleasant taste or odor, aromas, colors, added fragrances and substances to enhance the appearance of the composition. Acceptable excipients include stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, magnesium carbonate, talc, gelatin, acacia gum, sodium alginate, pectin, dextrin, mannitol, sorbitol, lactose, sucrose, starches, gelatin, cellulosic materials such as alkanoic acid cellulose esters and cellulose alkylesters, low melting wax, cocoa butter or powder, polymers such as polyvinylpyrrolidone, polyvinyl alcohol and polyethylene glycols and other acceptable pharmaceutical materials. The pharmaceutical composition components may be encapsulated or tableted for convenient administration.

For oral administration, the pharmaceutical composition may be in the form of, for example, tablet, capsule, suspension or liquid. If desired, other active ingredients may be included in the

Composição1551849 composition. The suspension or liquid may include other additives such as β cyclodextrins, such as Captisol®, which may act as a solubilizing agent.

Dragee cores are provided with suitable coatings. For this purpose concentrated sugar solutions may be used which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol and / or titanium dioxide, lacquer solutions and suitable organic solvents or mixtures. of solvent. Dyestuffs or pigments may be added to the tablets or dragee coatings to identify or characterize different combinations of active compound doses.

Pharmaceutical compositions which may be used orally include snap caps made of gelatin as well as soft sealed caps made of gelatin and a plasticizer such as glycerol or sorbitol. Snap-in capsules may contain the active ingredients mixed with a filler such as lactose, a binder such as starch and / or a lubricant such as talc or magnesium stearate and optionally stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids such as fatty oils, liquid paraffin, liquid polyethylene glycols, Cremophor, Capmul, mono-, di- or long chain triglycerides. To these formulations can also be added stabilizers.

ΡΕ1551849

Liquid form compositions include solutions, suspensions and emulsions. For example, solutions of the compounds of this invention dissolved in water and water-propylene glycol and water-polyethylene glycol systems, optionally containing suitable conventional coloring agents, flavoring agents, stabilizers and thickening agents, may be provided.

Alternatively, the compound of formula I may be in powder form for constitution with a suitable carrier, for example sterile, pyrogen-free water, prior to use.

For administration in suppositories, the compounds may also be formulated by mixing the agent with a non-irritable suitable excipient which is solid at room temperature but liquid at rectal temperature and which will therefore melt into the rectum to release the drug. These materials include cocoa butter, beeswax and other glycerides.

As a topical treatment, an effective amount of Formula I is mixed in a pharmaceutically acceptable gel or cream carrier that can be applied to the patient's skin in the treatment area. The preparation of such creams and gels is well known in the art and may include penetration enhancers, such as oils or alcohols, which increase or allow the compounds of formula I to penetrate the dermis to the transdermal tissue.

ΡΕ1551849

In certain embodiments, the compound of formula I may be administered by inhalation as long as the compounds pass into the bloodstream. For example, pharmaceutical compositions containing the compound of formula I may conveniently be delivered by aerosol spray in the form of solution, dry powder or cream. The aerosol may use a pressurized package or a nebulizer and a suitable propellant. In the case of a pressurized aerosol, the metering unit may be controlled by means of a valve to deliver a metered amount. Capsules and cartridges of, for example, gelatin for use in an inhaler may be formulated containing a powder base such as lactose or starch.

Additionally, the compound of formula I may be delivered using a sustained release system. Various sustained release materials have been established and are well known to those skilled in the art. Sustained release capsules may, depending on their chemical nature, release the compounds for 24 hours to several days. Depending on the chemical nature and biological stability of the therapeutic reagent additional strategies for protein stabilization may be used.

The compound of formula I may also be delivered by controlled release formulation as well as may be provided in a dispersion of the active compound in hydroxypropyl methylcellulose or by other methods known to those skilled in the art.

ΡΕ1551849

The pharmaceutical compositions may also be part of a combination therapy. In a combination therapy, the compound of formula I and other drugs, such as other antimicrobial, anti-inflammatory and analgesic agents, may be administered simultaneously or at separate intervals. When administered simultaneously the compound of formula I and other drugs may be incorporated into a single pharmaceutical composition or in separate compositions, for example the compound of formula I in one composition and the other drugs in another composition. Each of these compositions may be formulated with common excipients, diluents or carriers and compressed into tablets or formulated into elixirs or solutions. The compounds may be formulated as sustained release dosage forms and the like.

When administered separately, therapeutically effective amounts of the compound of formula I and the other drugs are administered in a different program. One may be administered before the other as long as the time between the two administrations falls within a therapeutically effective range. A therapeutically effective interval is a period of time that begins when any one of (a) compound of formula I or (b) other drugs is administered to a mammal and ends within the limit of the beneficial effect in treating the combination of (a) and (B) . The mixture of different compounds of formula I may also be administered simultaneously or together.

ΡΕ1551849

In certain embodiments, the antibacterial compounds are prodrugs of the compounds of formula I. The term prodrug denotes a derivative of a known direct acting drug that is transformed into the active drug by an enzymatic or chemical process. Prodrugs of the compounds of formula I are prepared by modifying functional groups present in the compound so that the modifications are cleaved either in routine or in vivo manipulation to the parent compound. Prodrugs include, but are not limited to, compounds of structure (I) wherein the hydroxy, amine or sulfhydryl groups are attached to any group which, when administered to the animal, cleaves to form respectively the hydroxyl, amino groups. or free sulfhydryl. Representative examples of prodrugs include, but are not limited to: acetate, formate and benzoate derivatives of alcohol and amine functional groups. See Notari, RE, Theory and Practice of Prodrug Kinetics, Methods in Enzymology, 112: 309-323 (1985); Bodor, N., Novel Approaches in Prodrug Design, Drugs of the Future, 6 (3): 165-182 (1981); and Budgaard, H., Design of Prodrugs: Bioreversible-Derivatives for Various Functional Groups and Chemical Entities, in Design of Prodrugs (H. Bundgaard, ed.), Elsevier, NY (1985).

The compounds of this invention may be synthesized by various methods known to those skilled in the art. Non-limiting examples of synthetic schemes for producing antibacterial agents are described below.

Scheme 1 shows a method of preparing the compounds of Formula I which are the subject of this invention. The benzaldehydes of structure 1, which are listed below in schemes 3-9, are reacted with a cyclic amine reagent 2 under a variety of reaction conditions to yield intermediate 3. Preferred conditions include reacting 1 with 2 in a suitable solvent such as N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO) or acetonitrile in the presence of a suitable base such as potassium carbonate, dipotassium hydrogen phosphate, N, N-dihydroxide. isopropylethylamine and the like and at a suitable temperature (typically 50 ° C to reflux temperature) until the reaction achieves a high conversion to the desired product 3. 0 Compound 3 may be purified by column chromatography over silica gel or by recrystallization. Alternatively, the raw material can often be used directly in the next step. For this purpose compound 3 is reacted with an active methylene reagent 4 (R¹ and R² they are both electron removal groups) in a suitable solvent such as methanol, butanol and the like and at a suitable temperature as room temperature to generate intermediates 5. Compounds 4 are commercially available or generally known and in the patent literature. For example, 1,2,6-thiadiazine-3,5 (2H, 6H) -dione 1,1-dioxide is described in Goya, P. et al., Heterocycles 1981, 16, 5-7 and Ochoa. , C. et al. J. Heterocycl. Chem. 1978, 15, 221-224. N, N-dihydroxy derivatives of barbituric acid are also known: Cowden, W. et al. Aust. J. Chem. 1982, 35, 795-797. Barbituric acid and thiobarbituric acid are commercially available starting materials.

ΡΕ1551849

Compound 5 may, in certain cases, be isolated and purified. However, it is often preferable simply to continue the reaction for longer periods and / or at a higher temperature (typically 50 ° C to reflux temperature) to increase the reaction and / or to complete to produce the target and claimed compounds 6, which are compounds of Formula I. It will be apparent to one skilled in the art that the above synthetic description is merely representative and that additional non-essential variations known to one skilled in the art are possible, some of which are encompassed by the examples of this invention.

Scheme 1

<img file="PT1551849E_D0005.tif" />

(YF, C! 8f OTf)

<img file="PT1551849E_D0006.tif" />

<img file="PT1551849E_D0007.tif" />

<img file="PT1551849E_D0008.tif" />

.....—......►

<img file="PT1551849E_D0009.tif" />

s

<img file="PT1551849E_D0010.tif" />

(Formula I,

ΡΕ1551849

In the case of spirocyclic hydantoin derivatives of the compounds of Formula I (R¹ and R² taken together to form a hydantoin ring), the desired compounds may be prepared as indicated in Scheme 2. Ketones 7 (see below for illustrative examples) are reacted with potassium cyanide and ammonium carbonate or other equivalents. described in the following references to produce hydantoin products 8, which are compounds of Formula I, which are the subject of this invention. 0 One skilled in the art will appreciate that the foregoing synthetic description is merely representative and that additional non-essential variations known to one skilled in the art are possible, some of which are encompassed in the examples of this invention. References to this chemistry can be found in: Obrecht, D. et al., Helvetica Chimica Acta 1992, 75, 1666-96, Horn, E. et al., Chem. Ind. 1986, 61516, Grunewald, GL et al., J. Med. Chem. 1980, 23, 754-8 Denyer, CV et al., Bioorg. Med. Chem. Lett. 1992, 2, 1039-42, and references cited within the scope of these citations.

Scheme 2

<img file="PT1551849E_D0011.tif" />

<img file="PT1551849E_D0012.tif" />

Schemes 3-9 illustrate the general schemes of

Synthesis of 2-halobenzaldehydes. In addition to the large number of 2halobenzaldelds that are commercially available, one skilled in the art can produce several additional 2-halobenzaldehydes by one or both of the methods below.

With respect to Scheme 3, a 2-halotoluene may be oxidized to 2-halobenzaldehyde with chromium trioxide or similar oxidant in the presence of acetic anhydride or other acylating agent. See, for example, Org. Syn., Coll. Vol. 2, 1941, 441. Intermediate diacetate may be isolated and purified or may be hydrolyzed under acidic or basic conditions to the desired aldehyde. In addition to the many commercially available 2-halotoluenes that can be used, many more can be readily prepared by electrophilic aromatic substitution reactions (Friedel Crafts acylation, nitrization, chlorosulfonation, etc.) starting from 2-fluorotoluene. All four regio isomers of the products may be formed, in varying amounts, from these electrophilic aromatic substitution reactions.

SCHEME 3

Synthesis of 2-halobenzaldehydes by oxidation

<img file="PT1551849E_D0013.tif" />

CfO_The

AcgO

O

<img file="PT1551849E_D0014.tif" />

ΡΕ1551849

Referring to Scheme 4, a fluorobenzene may be lithium with lithium diisopropylamide, or other appropriate base, and the lithium salt may be retained with dimethylformamide or other appropriate electrophile to produce the desired aldehyde.

SCHEME 4

Synthesis of 2-fluorobenzaldehydes by lithium

F

<img file="PT1551849E_D0015.tif" />

In many cases where the above methods are not suitable for the synthesis of the necessary aldehydes, they may be made from the corresponding bromofluorobenzaldehydes as shown in Scheme 5. The aldehyde is first protected by acetalization with a 1.2 appropriate diol in the presence of an appropriate acid. The resulting compound is then treated with an appropriate metal reagent such as n-BuLi or Mg. 0 The resulting anion is quenched with the appropriate electrophile, and the aldehyde is deprotected with aqueous acid. A range of electrophiles may be used including Weinreb amides, aldehydes, ketones and disulfides.

SCHEME 5

Bromofluorobenzaldehydes

ΡΕ1551849

<img file="PT1551849E_D0016.tif" />

/

Η

Ηθ7

<img file="PT1551849E_D0017.tif" />

<img file="PT1551849E_D0018.tif" />

The required 4-Bromo-2-fluorobenzaldehyde and 5bromo-2-fluorobenzaldehyde are marketed by Aldrich Chemical Company as well as other suppliers. 2-Bromo-6-fluorobenzaldehyde may be made according to Scheme 3 starting from commercial 2-bromo-6-fluorotoluene. 3-Bromo-2-fluorobenzaldehyde may be made according to Scheme 4 starting from 2-fluorobromobenzene.

In certain cases, it may be advisable to introduce the amino substituent before introducing the electrophilic derivative substituent as shown in Scheme 6. Appropriate bromo-2-fluorobenzaldehyde is treated with the desired amine in the presence of an appropriate base such as potassium carbonate. . The 2-amino-bromo-benzaldeldo is then protected by acetalization with an appropriate 1,2-diol in the presence of an appropriate acid. 0 The resulting compound is then treated with an appropriate metal reagent such as nBuLi or Mg. The resulting anion is quenched with the appropriate electrophile and the aldehyde is deprotected with aqueous acid. A range of electrophiles may be used including Weinreb amides, aldehydes, ketones and disulfides.

ΡΕ1551849

SCHEME. 6th

<img file="PT1551849E_D0019.tif" />

In scheme 7, the desired benzaldehyde is formed by reduction and oxidation reactions. For example, o-fluoroaryl nitriles are reduced to the desired aldehydes (See A) by the methods described in Milos

Hudlicky, Reduction in Organic Chemistry, ACS Monograph 188, 2^The ed. 1996, p. Alternatively, the ofluorocarboxylic acid or ester moiety is reduced to the corresponding alcohol as described (Hudlicky, 1996), and then oxidation to the desired aldehyde (see B) by one of the many methods described in Milos Hudlicky, Oxidation in organic chemistry, ACS Monograph 186, 1990, 115-118,123126.

ΡΕ1551849

Scheme 7

Reduction Reactions

<img file="PT1551849E_D0020.tif" />

Formulation of fluoroarenes is shown in Scheme 8. Formulation of fluoroarenes can be achieved either by (a) direct metallization of the ortho position followed by retention with DMF, see Tetrahedron Lett. 1992, 33, 7499, and similar pathways in Bioorganic &amp; Medicinal

Chemistry, 1993, 6, 403, and J. Org. Chem. 1988, 53, 3145-7;

(b) metal halogen exchange followed by retention with DMF as described in Perkin 1, 2000, 24, 4234; (c) Lewis acid catalyzed formylation, see J. Med. Chem., 1988, 31, 1972-7 and J. Org. Chem., 1986, 51, 4073-5;

or by (d) Vilsmer type formylation, see J. Med. Chem., 1986, 29, 2250.

ΡΕ1551849

7

Scheme 8

Fluoroarenes Formulation

The)

<img file="PT1551849E_D0021.tif" />

&lt;b&gt;

, Br

T

<img file="PT1551849E_D0022.tif" />

(ç)

<img file="PT1551849E_D0023.tif" />

(Scheme 9 refers to methods for preparing aryl, vinyl, and amino substituted fluorobenzaldehydes, such as 1) Palladium-catalyzed aryl-aryl couplings described by N. Miyaura and A. Suzuki in Palladium-catalysed cros coupling reactions of organoboron compounds . Chem. Rev. 1995, 95, 2457-83; 2) arylamino couplings described by Buchwald et.al. in J. Am. Chem. Soc., 1994, 116, 7901, J. Am. Chem. Soc., 1996, 118, 7215, or Acc. Chem. Res. 1998, 805, or by Hartwig, et. al. in J. Am. Chem. Soc.,

1994, 116, 5969; 3) aryl vinyl couplings (reaction of

Heck) described in Angew. Chem., Int. Ed. Eng. 1995, 34,

1844, 1848 or in Tetrahedron Lett. 1996, 37, 6535; or 4) aryl-CO bond formation described in Tetrahedron Lett.

1986, 27, 3931.

ΡΕ1551849

Aryl, vinyl and amino substituted o-fluorobenzaldehydes

1) aryl-aryl palladium couplings

O

<img file="PT1551849E_D0024.tif" />

<img file="PT1551849E_D0025.tif" />

<img file="PT1551849E_D0026.tif" />

3) aryl vinyl couplings (reaction of

Heck):

ΡΕ1551849

4) aryl-CO bond formation

<img file="PT1551849E_D0027.tif" />

The compounds of Formula I include at least one chiral center. Formula I covers both racemic and enantiomerically enriched forms of the compound of this invention. The racemic mixture is useful in the same manner and to the same effect as the most active enantiomer; The difference is that it is necessary to use more racemic material to produce the same antibacterial effect. 0 One skilled in the art will appreciate that some of the claimed compounds have multiple chiral centers present. In these cases, diastereomers are possible.

All of these diastereomers, in racemic and enantiomerically enriched forms, are also within the scope of the claimed Formula I compounds.

Enantiomerically enriched term means that an enantiomer of a specific compound is present in a mixture of the enantiomers for that compound in an amount greater than the other enantiomer. For example, an enantiomerically enriched form may include a mixture of enantiomers of a specific compound wherein the concentration of a single enantiomer of that compound is greater than 50%, more typically higher.

ΡΕ1551849 at 60%, 70%, 80% or 90% or more relative to the other enantiomer of that compound.

Racemic examples of compounds of Formula I may be separated into individual enantiomers or enantiomerically enriched isomers by high pressure liquid chromatography (HPLC) over various chiral stationary phases. For example, chromatography of racemic material on a ChiralPack AD column with ethanol (0.1% DEA) or ethanol / isopropanol (0.1% DEA) yields an enantiomerically enriched material. Alternatively, racemic compounds may be separated into preparative HPLC enantiomerically enriched isomers using a Chirose C3 column and ethanol / isopropanol (0.1% DA). It should be noted that these are merely representative conditions and that other mobile and stationary phases are useful for producing enantiomerically enriched compounds of

Formula I.

Alternatively, the enantiomerically enriched compounds of Formula I may be prepared starting with enantiomerically enriched cyclic amines 2 (see Scheme 1).

Also, enantiomerically enriched compounds of Formula I may be prepared by crystallization of racemic mixtures in the presence of an enantiomerically enriched acid or base to produce a diastereomeric salt.

ΡΕ1551849

EXAMPLES

Without further elaboration, it is believed that one skilled in the art using the foregoing description may practice the present invention to its fullest extent. The following detailed examples describe how to prepare the various compounds and / or perform the various processes of the invention and are to be construed as illustrative only and not as limitations of the foregoing disclosure in any way. Those skilled in the art will readily recognize appropriate variations of procedures with respect to both reagents, reaction conditions and techniques.

EXAMPLE 1

1,2,3,4a-Tetrahydro-cis-2,4-dimethyl-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H ) -pyrimidine] 2 ', 4', 6 '(1'H, 3'H) -trione (12) and 1,2,4,4a-Tetrahydro-trans2,4-dimethyl-8-nitrospiro [[ 1,4] oxazine [4,3-a] quinoline 5 (6H), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

<img file="PT1551849E_D0028.tif" />

Step 1

Preparation of Cis- and trans-2- (2,6-dimethylmorpholin-4-yl) -5nitrobenzaldehyde

ΡΕ1551849

<img file="PT1551849E_D0029.tif" />

, CHO

A vial of 2-fluoro-5-nitrobenzaldehyde (1.00 g, 5.92 mmol), 2,6-dimethylmorpholine (0.796 g, 6.92 mmol) of anhydrous powdered potassium carbonate (0.955 g, 6 , 92 mmol) and dimethylformamide (5 mL). The mixture is heated to reflux for 1 h, cooled to room temperature and poured into 50% saturated saline (50 mL). The aqueous layer is extracted with ether (2x). The combined organic extracts are dried (Na2SO4).₄) and concentrates. The resulting residue is purified by silica gel chromatography using acetone methylene chloride heptane (0.5: 3: 6.5) as the eluent yielding the major cis isomer (1.18 g, 76%) as a yellow solid. and the smallest trans isomer (0.257 g, 16%) as a yellow oil. Cis: NMR of⁴H (400 MHz, CDCl1)₃) δ 10.09, 8.63, 8.32, 7.08, 3.93, 3.33, 2.84, 1.27.

Trans: NMR of ⁴H (400 MHz, CDCl1)₃) δ 10.10, 8.57, 8.27,

7,06, 4,26, 3,32, 3,10, 3,07, 1,31.

Step 2

Preparation of 5- [2- (cis-2,6-dimethylmorpholin-4-yl) -5-nitrobenzylidene] pyrimidine-2,4,6 (1H, 3H, 5H) -trione (9) and trans-5- [2- (2,6-dimethylmorpholin-4-yl) -5nitrobenzylidene] pyrimidine-2,4,6 (1H, 3H, 5H) -trione (10)

ΡΕ1551849

<img file="PT1551849E_D0030.tif" />

A mixture of cis-2- (2,6-dimethylmorpholin-4-yl) 5-nitrobenzaldehyde (from Step 1) (0.528 g, 2.00 mmol) and barbituric acid (0.256 g, 2.00 mmol) in methanol ( 20 mL) are stirred at room temperature for 24 h. The reaction mixture is adsorbed onto silica gel (10 g) on ​​a rotary evaporator keeping the water bath &lt;30 degrees and then purified on silica gel using an acetone-chloroform-acetic acid gradient solvent system (1: 9: 0.5%) initially and then acetone-chloroform-methanol-acetic acid (1: 8.5: 0.5: 0.5%) as the eluent to obtain 421 mg (56%) of 5- [2 - (cis-2,6-dimethylmorpholin-4-yl) -5-nitrobenzylidene] pyrimidine-2,4,6 (1H, 3H, 5H) -trione as an orange solid. NMR of⁴H (400 MHz, DMSO-ch) δ 11.38, 11.27, 8.75, 8.23, 8.01, 7.21, 3.74, 3.29, 2.69, 1.11.

Similarly, the trans minor trans2- (2,6-dimethylmorpholin-4-yl) -5-nitrobenzaldehyde isomer (from step 1) is converted to trans-5- [2- (2,6-dimethylmorpholin-4-yl). yl) 5-nitrobenzylidene] pyrimidine-2,4,6 (1H, 3H, 5H) -trione in 50% yield. NMR of⁴H (400 MHz, DMSO-ch) δ 11.38, 11.27, 8.65, 8.23, 8.08, 7.21, 4.06, 3.18, 2.95, 2.92,

1,22.

ΡΕ1551849

Step 3

Preparation of 1,2,4,4a-Tetrahydro-cis-2,4-dimethyl-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2 'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione (12) and 1,2,4,4a-Tetrahydrotrans-2,4-dimethyl-8- nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -triona.

5- [2- (cis-2,6-Dimethylmorpholin-4-yl) -5-nitrobenzylidene] pyrimidine-2,4,6 (1H, 3H, 5H) -trione (from Step 2) (2.57 g, 6.87 mmol) is refluxed in methanol (230 mL) for 6.25 h and then the reaction mixture is stirred at RT overnight. The bright yellow precipitated solid is isolated by filtration and dried under high vacuum at 120 ° C for 3 days to yield 1,2,4,4a-tetrahydro-cis-2,4-dimethyl-8-nitrospiro [[1,4] ] oxazine [4,3-a] quinoline 5 (6H), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione in near quantitative yield . NMR of⁴H (400 MHz, DMSO-cG) δ 11.88, 11.57, 7.97, 7.04, 4.30, 3.92, 3.54, 2.99, 2.87,

I, 16, 0.94.

Similarly, trans-5- [2- (2,6-dimethylmorpholin-4-yl) -5-nitrobenzylidene] pyrimidine-2,4,6 (1H, 3H, 5H) trione (from Step 2) is converted to 1,2,4,4a-tetrahydrotrans-2,4-dimethyl-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2Ή) -pyrimidine] -2 ', 4', 6 '(1H, 3'H) -trione in 43% yield. NMR of⁴H (400 MHz, DMSO-cG) δ 11.88,

II, 60, 7.98, 7.83, 6.85, 6.85, 4.19, 4.07, 3.95, 3.89,

3,63, 3,56, 2,91, 1,24, 0,91.

ΡΕ1551849

Example 2

1,2,4,4a-Tetrahydro-cis-2,4-dimethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H) pyrimidine] - 2 ', 4', 6 '(1'H, 3'H) -trione

<img file="PT1551849E_D0031.tif" />

Step 1

Preparation of Cis-2- (2,6-dimethylmorpholin-4-yl) benzaldehyde

<img file="PT1551849E_D0032.tif" />

Similarly to that described in Example 1 (Step 1) cis-2- (2,6-dimethylmorpholin-4-yl) benzaldehyde is obtained in 42% yield as a yellow solid. NMR of⁴H (400 MHz, CDCl1)₃) δ 10.33, 7.83, 7.55, 7, 13, 3.94, 3.10, 2.67, 1.25.

Step 2

Preparation of 1,2,4,4a-Tetrahydro-cis-2,4-dimethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H) - pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione.

Similar to what is described in Example

51551849 (Steps 2 and 3) to 1,2,4,4a-tetrahydro-cis-2,4-dimethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 ' (2'H) -pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione is obtained by a single vessel reaction (Steps 3 and Example 1) to form directly the spirocyclic end product at room temperature in 53% yield as a white solid. NMR of⁴H (300 MHz, DMSO-Ch) δ 11.69, 11.43, 7.06, 6.85, 6.54,

4,02, 3,65, 3,59-3,48, 3,23, 2,90, 2,78, 1,14, 0,91.

Example 3

8-Bromo-1,2,4,4a-tetrahydro-cis-2,4-dimethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H ) - pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione

<img file="PT1551849E_D0033.tif" />

Step 1

Preparation of Cys and trans-5-bromo-2- (2,6-dimethylmorpholin-4-yl) benzaldehyde

Similarly to that described in Example 1 (Step 1) cis -dehyde and trans-5-bromo-2- (2,6-dimethylmorpholin) cis-5-bromo-2- (2,6-dimethylmorpholin-4-yl) benzalΡΕ1551849 -4-yl) benzaldehyde are obtained in 63% and 16% yield as a yellow solid and a yellow oil, respectively. Cis isomer: NMR of⁴H (400 MHz, CDCl1)₃δ 10.25, 7.91, 7.62, 6.99, 3.96-3.86, 3.04, 2.65, 1.24; Trans isomer:⁴H (400

MHz, CDCl3) δ 10.33, 7.91, 7.63, 7.00, 4.28-4, 18, 3.11,

2, 80, 2, 77, 1,34.

Step 2

Preparation of 8-Bromo-1,2,4,4a-tetrahydro-cis-2,4-dimethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2 'H) -pyrimidine] -2', 4 ', 6' (1 ', 3') -trione.

Similarly to that described in Example 1 (Steps 2 and 3) to 8-bromo-1,2,4,4a-tetrahydro-cis-2,4-dimethylpiro [[1,4] oxazine [4,3- a] quinoline-5 (6H), 5 '(2'H) pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione is prepared in 88% yield as an orange solid. NMR of⁴H (400 MHz, DMSO-cW δ 11.78, 11.48, 7.18, 7.05, 6.81, 4.00, 3.67,

3,64-3,56, 3,56-3,46, 3,30, 2,86, 2,80, 1,13, 0,91.

Example 4

8-Fluoro-1,2,4,4a-tetrahydro-cis-2,4-dimethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H ) - pyrimidine] -2 ^ 4 ^ 6¹ (1'H, 3'H) -trione

<img file="PT1551849E_D0034.tif" />

ΡΕ1551849

Step 1

Preparation of 2- (2,6-dimethylmorpholin-4-yl) -5fluorobenzaldehyde

Similar to that described in Example 1 (Step 1) cis-2- (2,6-dimethylmorpholin-4-yl) -5-fluorobenzaldehyde and trans-2- (2,6-dimethylmorpholin-4-yl) - 5-Fluorobenzaldehyde are obtained in yields of 24% and 6% as a yellow solid and a yellow oil, respectively. Cis isomer: NMR of⁴H (400 MHz, CDCl1)₃) δ 10.37, 7.50, 7.31-7.24,

7.14, 7.12, 3.97-3.86, 3.00, 2.64, 1.24; and trans isomer:

NMR of ⁴H (400 MHz, CDCl1)₃) δ 10.45, 7.50, 7, 14, 7, 12, 4.244, 17, 3.07, 2.76, 2.74, 1.35.

Step 2

Preparation of 8-Fluoro-1,2,4,4a-tetrahydro-cis-2,4-dimethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2 'H) pyrimidine] -2', 4 ', 6' (1 ', 3') -trione.

Similar to that described in Example 1 (Steps 2 and 3) to 8-fluoro-1,2,4,4a-tetrahydro-cis-2,4-dimethylpiro [[1,4] oxazine [4,3- a] quinoline-5 (6H), 5 '(2') pyrimidine] -2 ', 4', 6 '(1', 3 ') -trione is prepared in 86% yield as a white solid. NMR of⁴H (400 MHz, DMSO-d6) δ 11.74, 11.48, 6.94-6.86, 6.86-6.75, 3.94,

3,66-3,49, 3,26, 2,92, 2,77, 1,13, 0,91.

ΡΕ1551849

Example 5

1,2,4,4a-Tetrahydro-cis-2,4-dimethyl-8trifluoromethyl spiro [[1,4] oxazine [4,3-a] quinoline5 (6H), 5 '(2'H) -pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione

<img file="PT1551849E_D0035.tif" />

Step 1

Preparation of Cys and trans-5-bromo-2- (2,6-dimethylmorpholin-4-yl) -5- (trifluoromethyl) benzaldehyde

<img file="PT1551849E_D0036.tif" />

Similar to that described in Example 1 (Step 1) cis-2- (2,6-dimethylmorpholin-4-yl) -5- (trifluoromethyl) benzaldehyde and trans-2- (2,6-dimethylmorpholin-4) -yl) -5- (trifluoromethyl) benzaldehyde are obtained in yields of 74% and 21% as yellow oils. Cis isomer: NMR of⁴H (400 MHz, CDCl1)₃) δ 10, 17.7, 98, 7.66, 7.07, 3.90-3.80, 3.09, 2.65, 1.17; Trans isomer:⁴H (400 MHz, CDCl 3) δ 10.32, 8.08, 7.76, 7.17, 4.32-4.22, 3.22, 2.93, 2.90, 1.37.

ΡΕ1551849

Step 2

Preparation of 1,2,4,4a-Tetrahydro-cis-2,4-dimethyl-8-trifluoromethyl spiro [[1,4] oxazine [4,3-a] quinoline5 (6H), 5 '(2') -pyrimidine ] -2 ', 4', 6 '(1', 3 ') -trione.

Similar to that described in Example 1 (Steps 2 and 3), except that after stirring at RT for 24 h the reaction mixture is heated at reflux for a further 3 h at 1.2 ° C. 4,4a-tetrahydro-cis-2,4-dimethyl8-trifluoromethyl spiro [[1,4] oxazine [4,3-a] quinoline 5 (6H), 5 '(2'H) -pyrimidine] -2', 4 ' , 6 '(1'H, 3'H) -trione is prepared in 53% yield as a white solid. NMR of⁴H (400 MHz, DMSO-d 6) δ 11.80, 11.51, 7.36, 7.21, 6.99, 4.16,

3,79, 3,65-3,55, 3,55-3,49, 3,45, 2,90, 1,15, 0,93.

Example 6

1,1 ', 2,3'4,4', 4a, 6'-Octahydro-cis-2,4 ', 6'-trioxospiro [[1,4] oxazine [4,3-a] quinoline- 5 (6ΈΓ), 5 '(2'H) pyrimidine] -8-carbonitrile

<img file="PT1551849E_D0037.tif" />

Step 1

Preparation of 4- (cis-2,6-dimethylmorpholin-4-yl) -3formylbenzonitrile

ΡΕ1551849

<img file="PT1551849E_D0038.tif" />

A mixture of 5-bromo-2- (2,6-dimethylmorpholin-4yl) benzaldeldo (0.298 g, 1.00 mmol), 1,1'-bis (dimethylphosphino) ferrocene [dppf] (0.022 g, 0.040 mmol), tris (dibenzylideneacetone) dipaladium [Pd₂(dba)₃] (0.018 g, 0.020 mmol), zinc powder (0.008 g, 0.12 mmol) and zinc cyanide (0.070 g, 0.60 mmol) in dimethylacetamide (2 mL) is heated to 120 ° C^TheC for 4 h. The reaction mixture is cooled and partitioned between dilute ammonium hydroxide and ethyl acetate. The phases are separated. The organic layer is extracted with an additional portion of ethyl acetate. The combined organic layers are dried (NaSO4) and then concentrated. The resulting residue is purified by silica gel chromatography using ethyl acetate-methyleneheptane chloride (0.5: 4.0: 5.5) as the eluent to yield 172 mg (70%) of cyano aldeldo. NMR of⁴H (400 MHz, CDCl3) δ 10.05, 7.97, 7.64, 7.01, 3.92-3.79, 3.14, 2.70, 1.17.

Step 2

Preparation of 1,1 ', 2,3'4,4a, 6'-Octahydro-2,4', 6'-trioxospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H ), 5 '(2'H) pyrimidine] -8-carbonitrile

Similar to that described in Example 1 (Steps 2 and 3) except that after stirring at RT for 36 h, the reaction mixture is heated to

ΡΕ1551849 for an additional 24 h to yield 1,1 ', 2,3'4,4', 4a, 6'-octahydro-2,4 ', 6'-trioxospiro [[1,4] oxazine [4, 3-a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] -8-carbonitrile in 88% yield as a white solid. NMR of⁴H (400 MHz, DMSO-cA) δ 11.90, 11.60, 7.53, 7.32, 7.06, 4.26, 3.88,

3,68-3,59, 3,59-3,51, 3,46, 2,97, 2,90, 1,20, 0,98.

Example 7

1,2,4,4a-Tetrahydro-cis-2,4-dimethyl-8carboxamide spiro [[1,4] oxazine [4,3-a] quinoline5 (6H), 5 '(2'H) -pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione

<img file="PT1551849E_D0039.tif" />

Step 1

Preparation of 4- (cis-2,6-dimethylmorpholin-4-yl) -3formylbenzamide

<img file="PT1551849E_D0040.tif" />

A solution of 4- (cis-2,6-dimethylmorpholin-4-yl) 3-formylbenzonitrile (Example 6, Step 1) (0.244 g, 1.00 mmol) in acetone (4 mL) is treated with potassium carbonate in anhydrous powder (0.028 g, 0.20 mmol) followed by 30% of

51551849 hydrogen peroxide (0.204 g, 6.00 mmol). The reaction mixture is stirred at room temperature. After 24h, an additional 30% hydrogen peroxide (6.0 mmol) and acetone (2 mL) are added. After stirring at RT for 5 days, additional potassium carbonate (0.96 g) and DNSO (4 mL) are added and stirring at RT for 24 h. The reaction mixture is poured into 50% saturated saline (40 mL), extracted with ether (3x), dried (NaSO4).₄) and concentrated. The resulting residue is purified by silica gel chromatography using acetone-methylene chloride (1: 4) as the eluent to afford 28 mg (11%) of the desired amide as a yellow foam. NMR of⁴H (400 MHz, CDCl1)₃) δ 10.20, 8.20, 8, 09, 7.12, 6.47-5.88, 4.00, 3.86, 3.20, 2.74,15.

Step 2

Preparation of 1,2,4,4a-Tetrahydro-cis-2,4-dimethyl-8carboxamide spiro [[1,4] oxazine [4,3-a] quinoline 5 (6H), 5 '(2 Ή) -pyrimidine ] -2 ', 4', 6 '(1', 3 ') -trione.

<img file="PT1551849E_D0041.tif" />

Similar to that described in Example 1 (Steps 2 and 3) to 1,2,4,4a-tetrahydro-cis-2,4-dimethyl-8carboxamide spiro [[1,4] oxazine [4,3- a] quinoline 5 (6H), 5 '(2Ή) -pyrimidine] -2', 4 ', 6' (1Ή, 3'H) -trione is obtained as a white solid in 90% yield after stirring at RT for 9 days. NMR of⁴H (400 MHz, DMSO-d 6) δ

ΡΕ1551849

11, 78, 11, 47, 7,64, 7,60, 7, 43, 6, 90, 6, 87, 4, 15, 3, 75,

3,62-3,42, 3,28, 2,93-2,81, 1,15, 0,93.

Example 8

1,2,4,4a-Tetrahydro-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

<img file="PT1551849E_D0042.tif" />

Step 1

Preparation of 2-morpholin-4-yl-5-nitrobenzaldehyde

<img file="PT1551849E_D0043.tif" />

Similarly to that described in Example 1 (Step 1) 2-morpholin-4-yl-5-nitrobenzaldehyde is obtained in 89% yield as a yellow solid. NMR of⁴H (400 MHz, CDCl1)₃) δ 10.05, 8.57, 8.26, 7.03, 3.87, 3.23.

Step 2

Preparation of 5- (2-morpholin-4-yl-5-nitrobenzylidene) pyrimidine-2,4,6 (1H, 3H, 5H) -trione

<img file="PT1551849E_D0044.tif" />

ΡΕ1551849

Similar to that described in Example 1 (Step 2) 5- (2-Morolinolin-4-yl-5-nitrobenzylidene) pyrimidine-2,4,6 (1H, 3H, 5H) -trione is obtained in a yield 15% as a yellow solid. NMR of⁴H (300 MHz, DMSO-d8) δ 11.40, 11.28, 8.71, 8.25, 8.04, 7.22, 3.74, 3.17.

Step 3

Preparation of 1,2,4,4a-Tetrahydro-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H) pyrimidine] -2', 4 ', 6' (1Ή, 3Ή) -trione.

Similar to that described in Example 1 (Step 3) at 1,2,4,4a-tetrahydro-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H) , 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione is obtained in a yield of 73%. NMR of⁴H (400 MHz, DMSO-cA) δ 11.63, 11.41, 7.96, 7.86, 7.04, 4.13,

3,91, 3,85, 3,76, 3,51,-3,38, 3,30-3,11.

Example 9

1,2,4,4a-Tetrahydro-2,4-dimethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] - 2 ', 4', 6 '(1'H, 3'H) -trione

H

<img file="PT1551849E_D0045.tif" />

ΡΕ1551849

Step 1

Preparation of 5- (2-morpholin-4-ylbenzylidene) pyrimidine2,4,6 (1H, 3H, 5H) -trione.

HN NH

<img file="PT1551849E_D0046.tif" />

Similarly to that described in Example 1 (Step 2) 5- (2-morpholin-4-ylbenzylidene) pyrimidine2,4,6 (1H, 3H, 5H) -trione is obtained in 35% yield as a solid. orange. NMR of⁴H (400 MHz, DMSO-cu) δ 11.31,

11, 12, 8,36, 7, 88, 7, 48, 7, 10, 7, 04, 3, 72, 2, 94.

Step 2

Preparation of 1,2,4,4a-Tetrahydro-2,4-dimethylpiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2Ή) pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

Similar to that described in Example 1 (Step 3) to 1,2,4,4a-tetrahydro-2,4-dimethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 ( 6H), 5 '(2') -pyrimidine] -2 ', 4',

6 '(1'H, 3'H) -trione is obtained in 93% yield after refluxing for 2 hours in methanol. NMR of⁴H (300 MHz, DMSO-Ch) δ 11.48, 11.35, 7.06, 6.97, 6.90, 6.69,

3,86, 3,70, 3,50, 3,41-3,08, 2,88.

ΡΕ1551849

- 67 Example 10

8-Bromo-1,2,4,4a-tetrahydro-2,4-dimethyl spiro [[1,4] piperazine [4,3-a] quinoline-5 (6H), 5 '(2'H) pyrimidine ] -2 ', 4', 6 '(1'H, 3'H) -trione.

<img file="PT1551849E_D0047.tif" />

Step 1

Preparation of 5-bromo-2- (3,5-dimethylpiperidin-lil) benzaldehyde

<img file="PT1551849E_D0048.tif" />

Similar to that described in Example 1 (Step 1) 5-bromo-2- (3,5-dimethylpiperidin-1-yl) benzaldehyde is obtained as a diastereomeric mixture in 85% yield. NMR of⁴H (400 MHz, CDCl 3) δ 10.30, 10.20, 7.90, 7.58, 7.00, 3.17, 3.06, 2.73, 2.70, 2.41,

2,19-2,09, 2,01-1,84, 1,62, 1,46, 1,06, 0,92.

Step 2

Preparation of 8-Bromo-1,2,4,4a-tetrahydro-2,4-dimethylpiro [[1,4] piperazine [4,3-a] quinoline-5 (6H),

5 '(2'H) -pyrimidine] -2', 4 ', 6' (1Ή, 3Ή) -trione.

ΡΕ1551849

Similar to that described in Example 1 (Steps 2 and 3) to 8-Bromo-1,2,4,4a-tetrahydro-2,4-dimethylpiro [[1,4] piperazine [4,3-a] quinoline-5 (6H),

5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione is obtained in 89% yield as a white solid. NMR of⁴H (400 MHz, DMSO-dc) δ 11.63, 11.49, 7.15, 7.03, 6.69, 3.87, 3.61,

3,25, 2,78, 2,66, 1,78, 1,55, 0,88, 0,65.

Example 11

1,2,4,4a-Tetrahydro-1,4a-dimethyl-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H) pyrimidine ] -2 ^ 4 ^ 6¹ (1'H, 3'H) -trione.

<img file="PT1551849E_D0049.tif" />

Step 1

Preparation of 3,5-dimethylmorpholine

<img file="PT1551849E_D0050.tif" />

Acetol (10.9 q, 0.133 mmol, 1.17 ea) and dI-2-amino-1-propanol (8.5 g, 0.113, 1.0 ea) are combined into

200 mL of ethanol. The atmosphere over the solvent is purged with nitrogen, platinum oxide (50 mg, 85%, Englehart) is added and the mixture is hydrogenated at room temperature.

ΡΕ1551849 at 38.5 psi at night. An additional 50 mg of platinum oxide catalyst is added and the reaction mixture is hydrogenated for an additional 18 h. The reaction mixture is filtered through a Solka-Floc pad, carefully rinsed with ethanol and the filtrate concentrated. The residue is purified by silica gel chromatography using chloroform-methanol-ammonium hydroxide (29%) (85: 15: 1) as the eluent to yield 10.67 g (71%) of aminodiol.

Subsequent dehydration of aminodiol (10.1 g, 0.0759 mol) is carried out in a flask having a large empty volume to accommodate the foam generated by heating with concentrated sulfuric acid (14.14 g, 0.144 mol, 1.90 eq). at 180 ° C for 8 h. The black mixture is cooled in an ice bath while potassium hydroxide (17.1 g, 0.304 mol, 4.0 ea) in 85 mL of water is added dropwise over a period of 25 min. The basic suspension is stirred at room temperature overnight, filtered through a pad of celite and the pad is washed twice with water. The aqueous filtrate is extracted five times with chloroform-methanol (85:15), dried (NaSO4).₄) and is concentrated on a rotary evaporator, keeping the water bath temperature &lt;25 ° C to minimize loss of volatile product 7.19 g (82%) as a colorless liquid: 1 H NMR. ⁴H (400 MHz, CDCl 3) δ 3.62, 3.23, 3.20, 3.07, 2.89, 1.03, 0.88 3.

Step 2

Preparation of 2- (3,5-dimethylmorpholin-4-yl) -5nitrobenzaldehyde

ΡΕ1551849

Oj »N

<img file="PT1551849E_D0051.tif" />

Similar to that described in Example 1 (Step 1), 2- (3,5-dimethylmorpholin-4-yl) -5-nitrobenzaldehyde is obtained in 26% yield as a mixture of cis and trans isomers, which is used immediately in the next reaction without further purification. NMR of⁴H (400 MHz, DMSO-cW δ 11.63, 11.49, 7.15, 7.03, 6.69, 3.87, 3.61,

3,25, 2,78, 2,66, 1,78, 1,55, 0,88, 0,65.

Step 3

Preparation of 1,2,4,4a-Tetrahydro-1,4a-dimethyl-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H ) pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione.

Similarly to that described in Example 1 (Steps 2 and 3) 36 mg 1,2,4,4a-tetrahydro-1,4a-dimethyl-8-nitrospiro [[1,4] oxazine [4, 3-a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione are obtained as a yellow oil (mixture of cis and trans isomers) after stirring at room temperature for 6 days followed by purification on silica gel using acetate-chloroform-acetic acid (1: 2: 7: 0.5%) as the eluent. NMR of⁴H (400 MHz, DMSO-cA) δ 11.50, 11.36, 7.98, 7.94, 6.85, 3.97, 3.87.69, 3.67-3.47, 3.12, 1.29, 1.26.

ΡΕ1551849

Example 12

8-Bromo-1,2,4,4a-tetrahydro-cis-2,4-dimethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2 Ή) pyrimidine] -4'-thioxo-2 ', 6' (1'H, 3'H) -dione.

<img file="PT1551849E_D0052.tif" />

Similar to that described in Example (Steps 2 and 3), except that barbituric acid was replaced by thiobarbituric acid, 8Bromo-1,2,4,4a-tetrahydro-cis-2 is obtained. , 4-dimethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2') -pyrimidine] -4'-thioxo 2 ', 6' (1'H, 3 ' H) -dione in 82% yield as a yellow solid after stirring at RT for 24 h and then heating at reflux for 5.5 h. NMR of⁴H (400 MHz, DMSOd)₆) δ 12, 86, 12.61, 7, 19, 7, 09, 6.83, 4.01, 3.64, 3.653,53 3.53-3.42, 3.30, 2.91, 2 , 81, 1.13, 0.90.

Example 13

8-Bromo-1,2,4,4a-tetrahydro-cis-2,4-dimethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2 Ή) pyrimidine] -2 ', 4', 6 '(1'methyl, 3'methyl) -trione.

<img file="PT1551849E_D0053.tif" />

ΡΕ1551849

Similar to that described in Example (Steps 2 and 3), except that barbituric acid was replaced by 1,3-dimethylbarbituric acid, 1,2,4,4a-tetrahydro-cis-2 is obtained. , 4-dimethyl-8-bromo-spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'7) pyrimidine] 2', 4 ', 6' (1 ' methyl, 3'methyl) -trione in 40% yield as a white solid after stirring at RT for 24 h and then heating at reflux temperature for 5.5 h. NMR of⁴H (400 MHz, DMSO-Ch) δ 7.19, 6.98, 6.68, 98, 6.84, 4.00, 3.68, 3.63-3.52 3.50-3.39, 3 , 32, 3.23, 3.23, 3.09, 2.92,

2, 80, 1, 12, 0, 87.

Example 14

N- [1,1 ', 2,3', 4,4 ', 4a, 6'-Octahydro-2,4-dimethyl-2^,, 4 ', 6'-trioxospiro [[1,4] oxazine [4,3-a] quinolone-5 (6H), 5' (2'H) -

<img file="PT1551849E_D0054.tif" />

One bottle of Parr is charged with 1,2,4,4atetrahydro-cis-2,4-dimethyl-8-nitrospiro [[1,4] oxazine [4,3a] quinoline-5 (6H), 5 ' (2'H) -pyrimidine] -2 ', 4', 6 '(1'H, 3'H) trione (Example 1, Step 3) (203 mg, 0.542 mmol), acetic anhydride (0.4 mL, 4.24 mmol) and 10 wt% Pd / C (28 mg). Methanol (35 mL) is added and the resulting mixture is

ΡΕ1551849 stirred at room temperature under 40 psi H₂ for 18 hours. The mixture is filtered through a filter pad.

Celite and the pad is washed with methanol, 5% methanol in ethyl acetate and then 5% methanol in CH₂C1₂. The combined filtrates are concentrated under pressure and dried under high vacuum at room temperature for 18 hours. The crude product is purified by silica gel chromatography using 5% methanol in CH₂C1₂ as the eluent to yield 168.4 mg (80%) of the desired product. NMR of⁴H (DMSO-ch) δ 11.69, 11.40, 9.59, 7.21, 7, 15, 6, 76, 3.93,

3,61, 3,54, 3,20, 2,90, 2,72, 1,95, 1,12, 0,91.

Example 15 tert-Butyl 1,1 ', 2,3', 4,4 ', 4a, 6'-Octahydro-2,4-dimethyl2', 4 ', 6'-trioxospiro [[1,4] oxazine [4,3-a] quinolone 5 (6H), 5 '(2'H) -pyrimidine] -8-ylcarbamate.

<img file="PT1551849E_D0055.tif" />

One bottle of Parr is charged with 1,2,4,4atetrahydro-cis-2,4-dimethyl-8-nitrospiro [[1,4] oxazine [4,3a] quinoline-5 (6H), 5 '( 2'H) -pyrimidine] -2 ', 4', 6 '(1'H, 3'H) trione (Example 1, Step 3) (6.81 mg, 18.2 mmol), di-butyl dicarbonate. tert (8.6 g, 37.5 mmol) and 10 wt% Pd / C (694 mg). Methanol (750 mL) is added carefully and the resulting mixture is stirred at RT under 30 psi H₂ for 22 h. The reaction is filtered through a

ΡΕ1551849 Celite pad. The pad is washed with methanol, 5% methanol in EtOAc and 5% methanol in CH 2 Cl 2 (100 mL each). The filtrate is concentrated. The crude product is dissolved in hot EtOAc and a small amount of methanol and is recrystallized from heptane. After cooling to room temperature and then stirring at 0 ° C for 2 hours, the solid is collected by filtration and dried (20 torr, 60 ° C, 16 hours) to yield 5.67 g (70%) tert-butyl. 1,1 ', 2,3', 4,4 ', 4a, 6'-octahydro-2,4-dimethyl-2', 4 ', 6'trioxospiro [[1,4] oxazine [4,3 -a] quinolone-5 (6A), 5 '(2' Af) -pyrimidin] -8-ylcarbamate as a white solid which decomposes at 244 ° C. The mother liquors from this crystallization are concentrated and the residue is purified by silica gel chromatography using 5% methanol in CH 2 Cl 2 as the eluent to yield an additional 1.8 g (22%) of the title compound. NMR of⁴Η (DMSO-cu): δ 11.68, 11.42,

8, 92, 7, 05, 6, 72, 3,90, 3,61, 3,52, 3,17, 2, 88, 2,69, 1, 44, 1,12, 0,90.

Example 16

8-Amino-1,2,4,4a-tetrahydro-2,4-dimethylpiro [[1,4] oxazine [4,3-a] quinolone-5 (6H), 5 '(2'H) monohydrochloride ) pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione.

• HCi

<img file="PT1551849E_D0056.tif" />

NH

ΡΕ1551849

A suspension of tert-butyl 1,1 ', 2,3', 4,4 ', 4a, 6'octahydro-2,4-dimethyl-2', 4 ', 6'-trioxospiro [[1,4 ] oxazine [4,3-a] quinolone-5 (6H), 5 '(2'H) -pyrimidine] -8-ylcarbamate (from Example 15) (7.4 g, 16.65 mmol) in 250 mL of CH 2 Cl 2 at room temperature is pipetted with a solution of 4.0 N HCl in dioxane (20 mL, 80 mmol). The resulting slurry is stirred at room temperature for 2 days and is then concentrated. The residue is dissolved in hot 1: 1 EtOAc / methanol (-300 mL) and recrystallized from heptane. After cooling to room temperature and then stirring at 0 ° C for 30 minutes, the solid is isolated by filtration and dried (20 torr, 60 ° C for 18 h) to yield 6.31 g (99%) of the compound. of the title as a beige solid. NMR of⁴H (DMSO-d6) δ 11.78, 11.47, 9.83, 7.06, 6.92, 6.87, 4.03, 3.67, 3.60, 3.50,

3,41, 2,90-2,82, 1,13, 0,91.

Example 17

9-Bromo-1,2,4,4a-tetrahydro-2,4-dimethyl-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6ΈΓ), 5 '(2 'H) pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione.

<img file="PT1551849E_D0057.tif" />

ΡΕ1551849

- 76 Step 1

Preparation of 4-Bromo-2-fluoro-5-nitrobenzaldehyde:

<img file="PT1551849E_D0058.tif" />

Dropwise, nitric acid (25 mL) is added to a solution of 4-bromo-2-fluorobenzaldehyde (5.12 g, 25.2 mmol) in sulfuric acid (25 mL). The mixture is stirred for 2 hours and then poured over ice. The product is extracted into MTBE (100 mL). The organic layer is washed with saturated aqueous NaHCO 3, dried (Na 2 SO 4).₄), and evaporated yielding 5.98 g of yellow solid. NMR of⁴Η (DMSO-cG) δ 10.13, 8.49, 8.22.

Step 2

Preparation of 4-Bromo-2- (2,6-dimethylmorpholin-4-yl) -5nitrobenzaldehyde:

<img file="PT1551849E_D0059.tif" />

AT THE₂

A solution of cis-2,6-dimethylmorpholine (1.06 g,

9.20 mmol) in acetonitrile (40 mL) is added to a flask containing 4-bromo-2-fluoro-5-nitrobenzaldehyde (from Step 1)

15551849 (2.0 g, 8.06 mmol) and potassium carbonate (3.92 g, 28.4 mmol). The resulting slurry is stirred at room temperature for 63 hours. The mixture is poured into MTBE (100 mL) and washed with water (100 mL) followed by brine (100 mL). The organics are filtered through silica gel and concentrated to yield 1.76 g of a bright yellow solid. NMR of⁴H (DMSO-d_and) δ 9.93, 8.44, 7.53, 3.74-3.81,

3,48, 2, 78, 1, 12.

Step 3

Preparation of 9-Bromo-1,2,4,4a-tetrahydro-2,4-dimethyl-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2 'H) pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione.

A mixture of 4-bromo-2- (2,6-dimethylmorpholin-4yl) -5-nitrobenzaldehyde (from Step 2) (989 mg, 2.88 mmol) and barbituric acid (369 mg, 2.88 mmol) in isopropanol (15 mL) is sealed in a 20 mL scintillation vial and shaken at 85 ° C for 2.5 hours. The product is adsorbed onto silica gel and purified by silica gel chromatography using a gradient from 20% EtOAc in CH 2 Cl 2 to 25% EtOAc in CH₂C1₂ as an eluent. The product is isolated and dried (20 Torr, 100 ° C) to yield 512 mg of yellow solid. NMR of⁴H (DMSO-cU δ 11.90, 11.62, 7.79, 7.36, 4.32, 3.87,

3,47-3,63, 2,94, 2,81, 1,15, 0,94.

Example 18

8-Acetyl-1,2,4,4a-tetrahydro-2,4-dimethylpiro [[1,4] oxazine [4,3-a] quinoline-5 (6ΈΓ), 5 '(2'H) pyrimidine ] -2 ', 4', 6 '(1'H, 3'H) -trione.

ΡΕ1551849

<img file="PT1551849E_D0060.tif" />

Step 1

Preparation of 5-Bromo-2- (2,6-dimethylmorpholin-4yl) benzaldehyde:

<img file="PT1551849E_D0061.tif" />

A mixture of 5-bromo-2-fluorobenzaldehyde (16.0 g, 78.6 mmol), cis-2,6-dimethylmorpholine (9.32 g, 80.9 mmol) and potassium carbonate (15.0 g, 108 mmol) in DMF (75 mL) is heated in an oil bath at 120 ° C for 14 hours. The mixture is diluted with MTBE (250 mL) and washed with water (4 x 200 mL). (Add ethyl acetate as needed to maintain the homogeneity of the organic layer. The organics are filtered through a 1 µm plug of silica gel and evaporated to yield 21.5 g of brown solid. This material is approximately 90% pure as judged by HPLC and is used without further purification. NMR of⁴H (DMSO-ch) δ 10, 11, 7, 78, 7, 75, 7, 17, 3, 77-3.86, 3.12, 2.56,

1,11.

ΡΕ1551849

- 79 Step 2

Preparation of 4- [4-Bromo-2- (1,3-dioxolan-2-yl) phenyl] -2,6-dimethylmorpholine:

<img file="PT1551849E_D0062.tif" />

5-Bromo-2- (2,6-dimethylmorpholin-4-yl) benzaldehyde (from Step 1) (10.0 g, 33.5 mmol) is dissolved in toluene (50 mL), and sodium hydroxide monohydrate is added. toluenesulfonic acid (0.64 g, 3.4 mmol) and ethylene glycol (3.7 mL, 66.2 mmol). The reaction is refluxed for 2.5 hours with a Dean-Stark device in place. The reaction is neutralized with NaHCO₃ saturated and extracted with EtOAc (2x). The combined organic layers are washed with brine, dried (Na₂ONLY₄) and concentrated to yield a crude oil (13.09 g). The oil is purified by flash flash chromatography using an average 85 mm diameter sintered glass frit loaded to 50 mm with 15-40 microns SiO2. The product is eluted using a gradient of 99% heptane, 1% Et₃N to 90% ethylene chloride, 9% heptane, 1% Et₃N to produce 10.89 (95%) of golden oil. Y NMR (400 MHz, CDCl3₃) δ 7.66, 7.42, 6.94, 6, 13, 4, 18, 4.02, 3.83, 3.03, 2.46, 1.19.

Step 3

Preparation of 5-Acetyl-2- (2,6-dimethylmorpholin-4yl) benzaldehyde.

ΡΕ1551849

<img file="PT1551849E_D0063.tif" />

T-Butyllithium in hexanes (3.8 ml of a 1.7 M solution, 14.5 mmol) is added to THF at -78 ° C in a flame dried round bottom flask. 4- [4-Bromo-2- (1,3-dioxolan-2-yl) phenyl] -2,6-dimethylmorpholine (from Step 2) (1.0 g,

2.9 mmol) is dissolved in THF (5 mL) and slowly added to the t-BuLi solution over 7 minutes keeping the temperature below -70 ° C. The mixture is stirred for 30 minutes and N-methoxy-N-methyl acetamide (0.34 mL,

3.2 mmol) slowly, dropwise. The reaction is stirred at 78 ° C for 30 minutes and then stored at 0 ° C overnight. The reaction is quenched with 1M HCl (22.5 mL) and then heated at 65 ° C for 1 hour. The mixture is basified with Na₂CO₂ NaHCO 3 and extracted with EtOAc (2x). The combined organic layers are washed with brine, dried (Na₂ONLY₄) and concentrated. The resulting residue is adsorbed onto silica gel and purified by silica gel chromatography using 15-20% EtOAc in heptane as the eluent to yield 0.35 g (45%) of a clear oil. NMR of⁴H (400 MHz, CDCl 3) δ 10.1, 8.35,

8,12, 7,08, 3,93, 3,25, 2,76, 2,60, 1,25.

Step 4

Preparation of 8-Acetyl-1,2,4,4a-tetrahydro-2,4-dimethylpiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2ΈΓ) -pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione.

ΡΕ1551849

5-Acetyl-2- (2,6-dimethylmorpholin-4-yl) benzaldehyde (from Step 3, 0.34 g, 1.3 mmol) and barbituric acid (0.175 g, 1.36 mmol) are dissolved in isopropanol ( 6 ml) and heated at reflux for 1 hour. The cooled reaction mixture is stored at 0 ° C overnight. The pale yellow solid is isolated by filtration, washed with cold isopropanol and dried in a vacuum oven (20 Torr, 90 ° C) for 2 days to yield 0.48 g (100%) of the title compound. NMR of⁴H (400 MHz, DMSO-d c) δ 11.8, 11.45, 7.70, 7.52, 6.92, 4.36, 4.21, 3.82, 3.76, 3.60, 3.53, 3.43, 2.90,

2, 40, 1,15, 0, 92.

Example 19

8-Ethanone-O-Methyloxime-1,2,3,4a-tetrahydro-2,4-dimethyl spiro [[1,4] oxazine [4,3-a] quinoline-5 (6ΈΓ), 5 '(2JT ) - pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione.

<img file="PT1551849E_D0064.tif" />

8-Acetyl-1,2,4,4a-tetrahydro-2,4-dimethylpiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2H) pyrimidine] -2' , 4 ', 6' (1'H, 3'H) -trione as prepared in Example 18 (0.15 g, 0.40 mmol) and methoxyamine hydrochloride (0.17 g, 2.02 mmol) are dissolved in pyridine

51551849 (5 mL) and stirred at room temperature overnight. The reaction mixture is diluted with CH 2 Cl 2 and the solution is washed with NaHCO 3 and brine. The organic layer is dry (Na₂S)₄) and concentrated. The resulting crude solid is purified by silica gel chromatography using 5% IPA in CH₂C1₂ (150 mL) and 5% MeOH in CH₂C1₂ (100 mL) as the eluent to afford 120 mg (74%) of the title compound as a white solid. NMR of⁴H (400 MHz, DMSO-cL) δ 11.75, 11.45, 7.36, 6.84, 4.08, 3.84, 3.60, 4.53, 2.84,

2,07, 1,14, 0,91.

Example 20

1,2,3,4a-tetrahydro-2,4-dimethyl-8- (methylsulfonyl) spiro [[1,4] oxazine [4,3-a] quinoline-5 (6 #), 5 '(2ΈΓ ) pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione.

<img file="PT1551849E_D0065.tif" />

Step 1

Preparation of 4- [2- (1,3-oxolan-2-yl) -4- (methylthio) phenyl] 2,6-dimethylmorpholine:

<img file="PT1551849E_D0066.tif" />

ΡΕ1551849

To a stirred solution of 1.7 M t-butyllithium in hexane (11.4 mL, 19.0 mmol) in THF (35 mL) cooled to -78 ° C is added via addition for 15 min. a solution of 4- [4-bromo-2- (1,3-dioxolan2-yl) phenyl] -2,6-dimethylmorpholine (Example 18, Step 2) (3.0 g, 8, 8 mmol) in THF (15 mL). After stirring for 15 minutes, methyl methanethiol sulfonate (1.0 ml,

9.6 mmol) and the reaction is allowed to warm to 0 ° C. The reaction is stored at 0 ° C overnight. The reaction is poured into Na₂CO 3 and extracted with EtOAc (2x). The combined organic layers are washed with brine, dried (Na₂ONLY₄) and concentrated to yield 2.85 g (95%) of a golden oil which is used without further purification.

Step 2

Preparation of 2- (2,6-Dimethylmorpholin-4-yl) -5 (methylsulfonyl) benzaldehyde

<img file="PT1551849E_D0067.tif" />

4- [2- (1,3-Ioxolan-2-yl) -4- (methylthio) phenyl] -2,6-dimethylmorpholine (from Step I) (0.5 g, 1.62 mmol) is dissolved in a mixture of acetone (3 mL) and water (5 mL). Oxone (1.29 g, 2.1 mmol) is dissolved in 0.4 mM EDTA solution (5.3 mL) and slowly added to the reaction mixture. THE

The resulting suspension is vigorously stirred for one hour. The reaction is treated with a solution of NaHSCα (82 mg) in water (0.2 mL), followed by 1 M HCl (15 mL). The resulting mixture is heated at 65 ° C for 1 hour. The reaction is neutralized with Na₂Saturated aqueous CO 3 and extracted with EtOAc (2x). The combined organic layers are washed with brine, dried (Na₂ONLY₄) and concentrated. The resulting residue is adsorbed onto silica gel and purified by silica gel chromatography using 45-50% EtOAc in heptane as the eluent to yield 0.24 g (50%) of 2- (2,6-dimethylmorolinolin-4-yl). ) -5- (methylsulionyl) benzaldehyde as a yellow oil. NMR of⁴H (400 MHz, CDCl 3) δ 10.1, 8.31, 8.01, 7.15, 3.93, 3.25, 3.07, 2.79, 1.25.

Step 3

Preparation of 1,2,4,4a-Tetrahydro-2,4-dimethyl-8 (methylsulfonyl) spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H),

5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione.

Similar to that described in Example 18 (Step 4), 0.18 g (57%) of 1,2,4,4a-tetrahydro-2,4-dimethyl-8- (methylsulionyl) spiro [ [1,4] oxazine [4,3a] quinoline-5 (67%), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'7J, 3'H) trione. NMR of⁴H (300 MHz, DMSO-cA) δ 11.7, 7.55, 7.39,

4,21, 3,82, 3,60, 3,52, 3,04, 2,90, 1,15, 0,93.

Example 21

1,2,4,4a-Tetrahydro-2,4-dimethyl-8 (methylsulfinyl) spiro [[1,4] oxazine [4,3-a] quinoline-5 (6ΈΓ),

5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

ΡΕ1551849

<img file="PT1551849E_D0068.tif" />

Step 1

Preparation of 2- (2,6-Dimethylmorpholin-4-yl) -5 (methylsulfinyl) benzaldehyde

<img file="PT1551849E_D0069.tif" />

4- [2- (1,3-Ioxolan-2-yl) -4- (methylthio) phenyl] -2,6-dimethylmorpholine from Example 20 (Step 1, (1.6 g, 5.17 mmol) is dissolved. )) in a mixture of acetone (15 mL) and water (15 mL). The slurry is cooled to 0 ° C and NaHCO is added.₃ solid (3.79 g). To this stirred slurry is added dropwise a solution of Oxone (2.3 g) in 0.4 mM aqueous EDTA (10 mL) while maintaining the temperature below 5 ° C. The slurry is stirred for 5 minutes and then is treated with a NaHSO solution₃ (1.42 mg) in water (2.8 mL), followed by dioxane (10 mL) and 6 N HCl (9 mL). The reaction mixture is heated to 65 ° C for 1 hour. The reaction mixture is neutralized with Na₂CO₃ and is extracted with EtOAc (2x). The combined organic layers are washed with

ΡΕ1551849 saline solution, dried (Na₂ONLY₄) and concentrated. The crude oil is purified by silica gel chromatography using EtOAc as the eluent to yield 0.80 g (55%) of a yellow oil. NMR of⁴H (400 MHz, CDCl1)₃) δ 10.25, 7.98, 7.89,

7,23, 3,93, 3, 16, 2, 74, 2, 74, 1,25.

Step 3

Preparation of 1,2,4,4a-Tetrahydro-2,4-dimethyl-8 (methylsulfinyl) spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H),

5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione.

Similar to that described in Example 18 (Step 4) yields 0.85 g (77%) of 1,2,4,4a-tetrahydro-2,4-dimethyl-8- (methylsulfinyl) spiro [[ 1,4] oxazine [4,3a] quinoline-5 (6%), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) trione as a White solid. NMR of⁴H (400 MHz, DMSO-cA) δ 11.8, 11.5, 7.36, 7.23, 7.01, 4.14, 3.76, 3.60, 3.52,

3,42, 2,87, 2,63, 1,14, 0,92.

Example 22

1,2,4,4a-Tetrahydro-2,4-dimethyl-8- (methylthio) spiro [[1,4] oxazine [4,3-a] quinoline-5 (6 '), 5' (2 ' H) pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione

<img file="PT1551849E_D0070.tif" />

ΡΕ1551849

Step 1

Preparation of 2- (2,6-Dimethylmorpholin-4-yl) -5 (methylthio) benzaldehyde

<img file="PT1551849E_D0071.tif" />

4- [2- (1,3-Ioxolan-2-yl) -4- (methylthio) phenyl] -2,6-dimethylmorpholine (Example 20, Step 1, 0.5 g) is dissolved in dioxane (10 mL ) and 1 M HCl (10 mL). The mixture is heated at reflux for 15 minutes. The reaction is quenched in

Na 2 CO 3 and saturated with EtOAc (2x). The combined organic layers are washed with brine and then concentrated. The crude oil is adsorbed onto silica gel and is purified by silica gel chromatography using 10 to 20% EtOAc in heptane as the eluent to afford 270 g (59%) of 2- (2,6-dimethylmorpholin-4-yl). ) -5 (methylthio) benzaldehyde as a yellow oil. NMR of⁴H (400 MHz, CDCl1)₃) δ 10.3, 7.69, 7.45, 7.04, 3.90, 3.02, 2.63,

2,49, 1,22.

Step 2

Preparation of 1,2,4,4a-Tetrahydro-2,4-dimethyl-8 (methylthio) spiro [[1,4] oxazine [4,3-a] quinoline-5 (6/0, 5 '( 2'H) -pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione.

ΡΕ1551849

<img file="PT1551849E_D0072.tif" />

Similar to that described in Example 18 (Step 4) yields 0.31 g (84%) of 1,2,4,4a-tetrahydro-2,4-dimethyl-8- (methylthio) spiro [[ 1,4] oxazine [4,3-a] quinoline-5 (67J), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'7 ', 3'H) trione as a White solid. NMR of⁴Δ (400 MHz, DMSO-d8) δ 11.75, 11.45, 7.05, 6.90, 6.82, 3.99, 3.63, 3.60, 3.53,

3,25, 2, 87, 2, 77, 2,34, 1,13, 0, 91.

Example 23

1,2,4,4a-Tetrahydro-2,4-dimethyl-9-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6ΈΓ), 5 '(2'H) pyrimidine ] -2 ', 4', 6 '(1'H, 3'H) -trione

<img file="PT1551849E_D0073.tif" />

Step 1

Preparation of 2-Fluoro-4-nitrobenzaldehyde bis bis acetate.

<img file="PT1551849E_D0074.tif" />

ΡΕ1551849

2-Fluoro-2-nitrotoluene (10.0 g, 63.2 mmol), acetic acid (100 mL) and acetic anhydride (100 mL) are added to a 500 mL 3-neck round bottom flask and cooled. to -14 ° C with an ice-salt sea bath. Concentrated sulfuric acid (14.6 mL) is slowly added over 30 minutes keeping the temperature below 9 ° C. The reaction is warmed to 0 ° C and stirred for 1.5 hours. The reaction is poured into ice water (1000 mL) and stirred for 15 minutes. The dark green slurry is filtered and the resulting light green solid is washed with cold water. The solid is slurried with aqueous Na 2 CO 3, filtered, rinsed with cold water and cold EtOH to yield 6.93 g, (40%) of a white solid. NMR of⁴H (400 MHz, CDCl1)₃) δ 8.9, 7.99, 7, 93, 7, 75, 2, 16.

Step 2

Preparation of 2-Fluoro-4-nitrobenzaldehyde

<img file="PT1551849E_D0075.tif" />

2-Fluoro-2-nitrobenzaldehyde bis bis acetate (from Step 1, (6.9 g, 25.4 mmol)), EtOH (15 mL, water (15 mL) and concentrated sulfuric acid (1.4 mL) are mixed together and heated to reflux for 40 minutes The reaction mixture is filtered through Solka-flok and cooled to 0 ° C. The white solid precipitate is isolated by

181551849 filtration, washed with cold water and dried (20 Torr, 70 ° C) to yield 3.2 g (49%) of the desired aldehyde: 1 H NMR. ⁴H (400 MHz, CDCl3) δ 10.5, 8.16, 8.09.

Step 3

Preparation of 2- (2,6-Dimethylmorpholin-4-yl) -4-nitrobenzaldehyde

<img file="PT1551849E_D0076.tif" />

A stirred mixture of 2-fluoro-4-nitrobenzaldehyde (1.0 g, 5.9 mmol), K 2 CO 3 (2.05 g, 14.8 mmol) and cisdimethylmorpholine (0.75 g, 6.5 mmol) in CH₃CN (10 mL) is heated at reflux overnight. The reaction is cooled, diluted with water and extracted with CH 2 Cl 2 (2x). The combined organic layers are washed with brine, dried (Na₂ONLY₄) and concentrated. The resulting crude residue is adsorbed onto silica gel and purified by silica gel chromatography using 50-100% CH₂C1₂ in heptane as the eluent to yield 0.48 g (31%) of the desired product as an orange solid. NMR of⁴H (400 MHz, CDCl1)₃) δ 10.3, 7.93,

3,93, 3,14, 2, 75, 1,26.

Step 4

Preparation of 1,2,4,4a-Tetrahydro-2,4-dimethyl-9-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6ΈΓ), 5 '(2'H ) pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione.

ΡΕ1551849

Similar to that described in Example 18 (Step 4), 360 mg (59%) of 1,2,4,4α-tetrahydro-2,4-dimethyl-9-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H),

5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione are obtained as an orange solid. NMR of²Δ (300 MHz, DMSO-cA) δ 11.8, 11.5, 7.63, 7.39, 7.12, 4.18, 3.74), 3.63, 3.49, 3.17 , 2.92, 1.1, 17.0, 94.

Example 24

1,2,4,4a-Tetrahydro-2,4-dimethyl-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H) - pyrimidine] -2 ', 4', 6 '(1'methyl, 3'methyl) -trione and 1,2,4,4a-Tetrahydro-2,4-dimethyl-8-nitrospiro [[1,4] oxazine [ 4,3-a] quinoline-5 (6H), 5 '(2'H) pyrimidine] -2', 4 ', 6' (1'H, 3'methyl) -trione.

<img file="PT1551849E_D0077.tif" />

<img file="PT1551849E_D0078.tif" />

1,2,4,4a-Tetrahydro-cis-2,4-dimethyl-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2') is dissolved H) pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione of Example 1 (0.45 g, 1.16 mmol) in DMF (25 mL) and iodomethane ( 0.15 mL, 2.32 mmol). The reaction is cooled to 0 ° C and a solution of 1 M potassium t-butoxide in THF (1.8 mL, 1.8 mmol) is added dropwise over 5 minutes. The reaction is stirred for 90 minutes. The reaction is diluted with water.

1551849 and extracted with CH 2 Cl 2 (2x). The combined organic layers are washed with water (3x), dried (Na 2 SO 4).₄) and concentrated. The resulting crude residue is taken up on silica gel and purified by silica gel chromatography using 0.5-2% MeOH in CH 2 Cl 2 as the eluent to yield 0.20 g (33%) of 1,2,4,4- tetrahydro-2,4-dimethyl-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] -2', 4 ' , 6 '(1' methyl, 3'methyl) -trione as a yellow solid after drying at 55 ° C for 3 days (1 H NMR).⁴H (400 MHz, CDCl 3) δ 8.10, 7, 77.6, 75, 4.08, 3.76, 3.61, 3.42, 3.25, 3.19, 3.11, 2, 99,

1.26, 0.98). Further elution yielded 0.21 g (36%) of 1,2,4,4α-tetrahydro-2,4-dimethyl-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline 5 (67%), 5 '(2', 7) -pyrimidine] -2 ', 4', 6 '(1' H,

3'methyl) -trione (mixture of diastereomers) as a yellow solid after drying at 55 ° C for 3 days (1 H NMR). ⁴H (400 MHz, DMSO-d 6) δ 11, 8, 7, 98, 7, 77, 7, 05, 4.30, 3.94,

3,53, 3,21, 3,02, 2,98, 2,87, 1,16, 0,92).

Example 25

1,2,4,4a-Tetrahydro-4-methyl-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] 2 ', 4', 6 '(1'H, 3') -trione and 1,2,4,4a-Tetrahydro-2-methyl-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline5 (6 '), 5' (2'H) -pyrimidine] -2 ', 4', 6 '(1'H, 3' s) -trione.

<img file="PT1551849E_D0079.tif" />

<img file="PT1551849E_D0080.tif" />

ΡΕ1551849

Step 1

To a stirred solution of 2-methylmorpholine hydrochloride in DMF (5 mL) is added diisopropylethylamine (0.25 mL, 1.48 mmol) followed by 2-fluoro-5-nitrobenzaldehyde (210.1 mg, 1.24 mmol) and carbonate. solid potassium (198.8 mg, 1.44 mmol). The reaction mixture is heated at 120 ° C for 2 h. The reaction mixture is cooled and partitioned between ether and a half saturated saline solution. The phases are separated. The organic layer is extracted with ether (3x). The combined organic phases are dried (MgSO 4), filtered and concentrated. The resulting residue is dissolved in CH 2 Cl 2 and purified by silica gel chromatography using 20% ​​ethyl acetate in hexane as the eluent yielding 303.0 mg (1.21 mmol, 98%) of the desired product as a bright yellow solid.

Step 2

Similarly to that described in Example 1 (Steps 2 and 3), 255.1 mg (60%) of the desired product is obtained as a mixture of isomers. NMR of⁴H (400 MHz, DMSO-ch) δ 11, 9, 11, 75, 11.5, 11.35, 11.3, 7, 96, 7, 89, 7, 83, 7, 05, 6, 9, 4.28-3.11, 3.16, 2.89, 1, 17, 0.95.

Example 2 6

2,3,4,4a-Tetrahydro-1 ', 3,3'-trimethylspiro [1H-pyrazino] [1,2-a] quinoline-5 (6ΈΓ), 5' (2'H) -pyrimidine] 2 ', 4', 6 '(1' H, 3 'H) -trione

ΡΕ1551849

<img file="PT1551849E_D0081.tif" />

Similar to that described in Example 1, apart from 2-fluorobenzaldehyde, 1-methylpiperazine and 1,3-dimethylbarbituric acid, 190 mg (55%) of the title compound is obtained. NMR of²Δ (400 MHz, CDCL 3) δ 7.19, 7.04, 6.95, 6.80, 3.98, 3.62, 3.55, 3.39, 3.32, 3.10, 3, 04, 2.82, 2.57, 2.23, 2.21, 1.86.

Example 27

2,3,4,4a-Tetrahydro-3-methylspiro [1H-pyrazino] [1,2-a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] 2', 4 ', 6 '(1Ή, 3'H) -trione

<img file="PT1551849E_D0082.tif" />

Similar to what is described in Example

1, other than 2-fluorobenzaldehyde and 1-methylpiperazine, gives 278 mg (89%) of the title compound. NMR of⁴H (400 MHz, DMSO-d_and) δ 11.46, 11.28, 7.03,

6,95, 6, 89, 6, 66, 3,91, 3,30, 3,17, 3,11, 2, 80, 2, 75, 2,59,

2, 16, 1, 98, 1, 78.

ΡΕ1551849

Example 28

1,1-Dimethylethyl 1,1'2,3 ', 4', 4a, 6'-octahydro-8-nitro2 ^ 4 ^ 6¹ trioxospiro [3 / T-pyrazino [1,2-a] quinoline 5 (6H), 5 '(2'H) -pyrimidine] -3-carboxylate

<img file="PT1551849E_D0083.tif" />

Similar to that described in Example 1, except that from 1-Boc-piperazine, 1.37 g (63%) of the title compound is obtained. NMR of⁴H (400 MHz, DMSO-d 6) δ 11.59, 11, 41, 7, 95, 7, 90, 7, 01, 4, 12, 3.85,

3,80, 3,41, 3,17, 3,06, 3,01, 2,88, 1,40.

Example 2 9

1,1-Dimethylethyl-1,1'2,3 ', 4', 4a, 6'-octahydro-8-nitro2 ', 4', 6'-trioxospiro [3H-pyrazino [1,2-a] quinoline5 (6H), 5 '(2'H) -pyrimidine] -3-carboxylate

<img file="PT1551849E_D0084.tif" />

ΡΕ1551849

Similar to what is described in Example

I, other than 5-cyano-2-fluorobenzaldehyde and 1-Boc-piperazine, yields 3.59 g (83%) of 1,1-dimethylethyl-8-cyano-1,1,2,3 ', 4', 4a, 6'-octahydro-2 ', 4', 6'trioxospiro [3'-pyrazino [1,2-a] quinoline-5 (6%), 5 '(2' H) pyrimidine] -3-carboxylate. NMR of⁴H (400 MHz, DMSO-cb) δ

II, 45, 11.38, 7, 45, 7, 40, 6, 99, 4.05, 3.80, 3, 73, 3, 70,

3,27, 3,18, 3,10, 2,95, 2,78, 1,39.

Example 30 rei- (2 'R, 4' S, 4 'aR) -1,1'2,3', 4 ', 4a, 6'-Hexahydro-2', 4 'dimethyl-1,3- dioxospiro [2R-indene-2,5 '(6'H) [1,4] oxazine [4,3a] quinoline-8'-carbonitrile

<img file="PT1551849E_D0085.tif" />

Similar to that described in Example 1, except that starting from 5-cyano-2-fluorobenzaldehyde and indanedione, 112 mg (30%) of the title compound is obtained. NMR of⁴H (400 MHz, DMSO-cb) δ 8.03, 7.93, 7.53, 7.34, 6.91, 4.32, 4.26, 3.93, 3.65, 3.30, 3.13, 2.97, 1.09,

0,82 .

Example 31 rei- (2R, 4S, 4aR) -1,2,4,4a-Tetrahydro-2,4-dimethyl [1,4] oxazine [4,3-a] quinoline-5,5,8 ( 6H) tricarbonitrile

ΡΕ1551849

<img file="PT1551849E_D0086.tif" />

Similar to that described in Example 1, with the difference that starting from 5-cyano-2-fluorobenzene

<td>zaldehyde and</td><td>malononitrile, 93 are obtained</td><td>mg</td><td> (32%)</td><td>of</td><td>compound</td>

<td>of the title.</td><td>NMR of ⁴H (400 MHz, CDCl1)₃</td><td>) δ</td><td> 7,52,</td><td> 7,</td><td> 40, 6,97,</td>

<td> 4,39, 4,15,</td><td> 3,69, 3,66, 3,59, 3,53, 3,</td><td> 49,</td><td> 3,40,</td><td> 3,</td><td> 37, 1,68,</td>

1,29.

Example 32 rei- (2S, 4R, 4aS) -8-Bromo-1,2,4,4a-tetrahydro-2,4-dimethyl [1,4] oxazine [4,3-a] quinoline-5, 5 (6H) -dicarbonitrile

<img file="PT1551849E_D0087.tif" />

Similar to that described in Example 1, except that from 5-cyano-2-fluorobenzaldehyde and malononitrile, 1.26 g (54%) of the title compound is obtained. NMR of⁴H (400 MHz, DMSO-d8) δ 7.40, 3.37,

7, 11, 4,23, 3, 98, 3,86, 3,81, 3, 74, 3,55, 3,12, 1,54, 1, 13.

ΡΕ1551849

Example 33

2,3,4a-Tetrahydro-3-methyl-8-nitro-2'-thiospiro [1'pyrazino [1,2-a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] 4 ', 6' (1'3'H) -dione)

<img file="PT1551849E_D0088.tif" />

Similar to that described in Example 1, except that 1-methylpiperazine and thiobarbituric acid are used, 840 mg (81%) of the title compound is obtained. NMR of⁴H (400 MHz, DMSO-d8) δ 12.61, 12.49, 7.94, 7.88, 7.07, 4.18, 3.71, 3.40, 3.17, 3.11, 2.80, 2.65, 1.96, 1.78.

In other embodiments, the compounds of formula I may be synthesized as indicated in Scheme 10.

SCHEME 10

Alternative Synthetic Scheme

<img file="PT1551849E_D0089.tif" />

ΡΕ1551849

Example 34 rei- (2R, 4S, 4aS) -9- (4-Chlorophenyl) -1,2,4,4a-tetrahydro-2,4-dimethylpiro [1,4] oxazine [4,3-a] quinoline-1 5 (6H), 5 '- (2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

<img file="PT1551849E_D0090.tif" />

A mixture of 9-Bromo-1,2,4,4a-tetrahydro-2,4-dimethylpiro [[1,4] piperazine [4,3-a] quinoline-5 (6H),

5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione (0.245 mmol) [prepared according to Example 10 (Steps 1-3) with 4-bromo-2-fluorobenzaldehyde as the starting aldehyde], 4-chlorophenylboronic acid (0.27 mmol), sodium carbonate (0.61 mmol), tetrakis (triphenylphosphine) palladium (0) ( 0.013 mmmol) in THF (18 mL) and water (2 mL) is purged with nitrogen and heated to reflux for 18 h. The mixture is cooled to room temperature and diluted with ethyl acetate (20 mL) and filtered. The filtrate is concentrated and the resulting residue, a semi-solid, is purified on silica gel using 20% ​​ethyl acetate in dichloromethane as the eluent to afford 15 mg (15%) of the desired coupled product. Y NMR (400 MHz, Acetonad₆) δ 1, 10, 1, 26, 3.03, 3.1-3.3, 3, 7-3.85, 3.9, 4.05, 6.85, 6.96, 7.4, 7.5, 8.8, 9.1.

ΡΕ1551849

100

Example 35 rei- (2R, 4S, 4aS) -1,2,4,4a-Tetrahydro-2,4-dimethyl-9 - [e (trifluoromethoxy) phenyl] spiro [[1,4] oxazine [4, 3-a] quinoline5 (6 '), 5' - (2 ') -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

<img file="PT1551849E_D0091.tif" />

Similar to that described in Example 34, 34.19 mg (17%) of rei- (2R, 4S, 4aS) -1,2,4,4atetrahydro-2,4-dimethyl1-9 are obtained. - [4- (trifluoromethoxy) phenyl] spiro [[1,4] oxazine [4,3-a] quinoline-5 (6A), 5 '- (2'H) -pyrimidine] 2', 4 ', 6' (1'H, 3'H) -trione. NMR of⁴H (400 MHz, Acetone-cG) δ 1.07, 1.22, 2.99, 3.15, 3.16, 3.6-3.8, 3.85, 4.29-4.34, 6, 86, 6, 88, 7, 40, 7, 76.

Example 36 rei- (2R, 4S, 4aS) -1,2,4,4a-Tetrahydro-9- (methoxyphenyl) -2,4 dimethylpiro [[1,4] oxazine [4,3-a] quinoline-5 (6ΈΓ), 5 '- (2'H) pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

<img file="PT1551849E_D0092.tif" />

Similar to what is described in Example

ΡΕ1551849

101

34, 45 mg (45) of rei- (2R, 4S, 4aS) -1,2,4,4a-tetrahydro-9- (methoxyphenyl) -2,4-dimethyl spiro [[1,4] oxazine] are obtained. [4,3 - a] quinoline-5 (6H), 5 '- (2'H) -pyrimidine] -2', 4 ', 6' (1'J, 3'H) trione. NMR of⁴H (400 MHz, CD₃OD) δ 0.93, 1.2, 2.9, 3.15

3,7-3,80, 3,83, 6,8, 6,9, 6,95, 7,53.

Example 37 rei- (2R, 4S, 4aS) -9- (3-Chloro-4-fluorophenyl) -1,2,4,4a-tetrahydro-2,4-dimethyl spiro [[1,4] oxazine [4,3 -a] quinoline5 (6ΈΓ), 5 '- (2Ή) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

<img file="PT1551849E_D0093.tif" />

Similar to that described in Example 34, 45 mg (40%) of rei- (2R, 4S, 4aS) -9- (3-chloro-4 fluorophenyl) -1,2,4,4a- tetrahydro-2,4-dimethyl spiro [[1,4] o xazino [4,3-a] quinoline-5 (67%), 5 '- (2'7%) pyrimidine] -2', 4 ', 6 '(1Ή, 3Ή) -trione. NMR of⁴H (400 MHz, CD₃OD) δ 0.96, 1.26

2, 92, 3,15, 3,7, 3, 75-3,82, 4, 15, 6, 80, 6, 97, 7,3, 7,6

7,7.

Example 38 rei- (2R, 4S, 4aS) -1,2,4,4a-Tetrahydro-2,4-dimethyl-9- (3nitrophenyl) spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

ΡΕ1551849

102

<img file="PT1551849E_D0094.tif" />

Similarly to that described in Example 34, 49 mg (44%) of rei- (2R, 4S, 4aS) -1,2,4,4a-tetrahydro-2,4-dimethyl-9- ( 3-nitrophenyl) spiro [[1,4] oxazine [4,3a] quinoline-5 (67%), 5 '(2' H) -pyrimidine] -2 ', 4', 6 '(1' 7%, 3 ' H) trione. MS (EI) mlz 450 (M +), 451, 450, 365, 364, 335, 319,

277, 225, 204, 165.

Example 39 rel-4 [(2R, 4S, 4aS) -1,1 ', 2,3', 4,4 ', 4a, 6'-Octahydro-2,4-dimethyl-2', 4 ', 6' -trioxospiro [[1,4] oxazine [4,3-a] quinoline5 (6 '), 5- (2'H) -pyrimidin] -9-yl] benzonitrile.

<img file="PT1551849E_D0095.tif" />

Similar to that described in Example 34, 25 mg (24%) of rel-Q - [(2R, 4S, 4aS) -1, 1 ', 2,3', 4,4 ', 4a are obtained. 2,6'-octahydro-2,4-dimethyl-2 ', 4', 6'-trioxospiro [[1,4] oxazine [4,3-a] quinoline-5 (67%), 5 '- (2 (H) -pyrimidin] -9yl] benzonitrile. NMR of⁴H (400 MHz, CD 3 OD) δ 1.05, 1.25,

2, 90, 3,15, 3, 7-3,85, 4, 15, 6, 91, 7, 03, 7, 07, 7, 80.

ΡΕ1551849

103

Example 40 rei- (2R, 4S, 4aS) -1,2,4,4a-Tetrahydro-2,4-dimethyl-9- [4- (methylsulfonyl) phenyl] spiro [[1,4] oxazine [4, 3-a] quinoline5 (6H), 5 '- (2') -pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione

<img file="PT1551849E_D0096.tif" />

Similarly to that described in Example 34, 34.33 mg (28%) of rei- (2R, 4S, 4aS) -1,2,4,4atetrahydro-2,4-dimethyl-9 are obtained. - [4- (methylsulfonyl) phenyl] spiro [[1,4] oxazine [4,3-a] quinoline-5 (67%), 5 '- (2'H) -pyrimidine] 2', 4 ', 6' (1'H, 3'H) -trione. MS (EO) mlz 483 (M⁺) , 484, 483,

440, 398, 397, 368, 352, 312, 310, 204.

Example 41 rei- (2R, 4S, 4aS) -1,2,4,4a-Tetrahydro-2,4-dimethyl-9- (4pyridinyl) spiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] -2', 4 ', 6' ('H, 3'H) -trione

<img file="PT1551849E_D0097.tif" />

Similarly to that described in Example 34, 12 mg (12%) of rei- (2R, 4S, 4aS) -1,2,4,4a-tetrahydro-2,4-dimethyl-9- ( 4-pyridinyl) spiro [[1,4] oxazine [4,3ΡΕ1551849

104

a] quinoline-5 (6%), 5 '- (2'H) -pyrimidine] - 2', 4 ', 6' ('Η, 3' Η) -

<td>triona.</td><td>IR (reflectance</td><td>diffuse)</td><td> 2984, 2837,</td><td> 2705</td><td>(B) ,</td>

<td>2660 (b),</td><td>2649 (b), 26226</td><td>(b) 2611</td><td>(b) 1748,</td><td> 1722</td><td>(s) ,</td>

<td>1693 (s),</td><td> 1601, 1408, 1358,</td><td colspan="2">1350, 1214, cm^-1.</td><td></td><td></td>

Example 42 rel-Methyl (2R, 4S, 4aS) -1,1'-2,3 ', 4,4a, 6'-Octahydro-2,4-dimethyl-2', 4 ', 6'-trioxospiro [[ 1,4] oxazine [4,3-a] quinoline5 (6H), 5 '- (2'fl) pyrimidine] -9-carboxylate

<img file="PT1551849E_D0098.tif" />

A mixture of 9-Bromo-1,2,4,4a-tetrahydro-2,4-dimethylpiro [[1,4] piperazine [4,3-a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] -2 ', 4', 6 '(1'H, 3'H) -trione [prepared according to Example 10 (Steps 1-3) except that 4-bromo-2-fluorobenzaldehyde is used as the starting aldehyde], (1.47 mmol), tetrakis (triphenylphosphine) palladium (0.44 mmol), N, N, N-diisopropylethylamine (4.4 mmol), Dimethylformamide (8 mL) and methanol (2 mL) is charged with carbon monoxide (600 psi) in a steel pump and heated to 100 ° C for 72 h. The mixture is cooled to room temperature and the pump is evacuated. The mixture is diluted with methanol and filtered. The filtrate is concentrated to yield a residue which is purified by silica gel chromatography using 20% ​​ethyl acetate in CH 2 Cl 2 as the eluent to afford 26 mg (6%) of the desired product. NMR of⁴H

ΡΕ1551849

105 (400 MHz, Acetone) δ 1.05, 1.24, 3.0, 3.15, 3.45, 3.6,

3, 7, 3,85, 4, 15, 7, 06, 7,27, 7, 44.

Example 43 rel-Methyl (2R, 4S, 4aS) -1,1'-2,3 ', 4,4a, 6'-Octahydro-2,4-dimethyl-2', 4 ', 6'-trioxospiro [[1,4] oxazine [4,3-a] quinoline 5 (6H), 5 '- (2'fl) pyrimidine] -8-carboxylate

<img file="PT1551849E_D0099.tif" />

Similar to that described in Example 42, except that 8-Bromo-1,2,4,4a-tetrahydro-2,4-dimethyl spiro [[1,4] piperazine [4,3-a ] quinoline5 (6H), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione (from Example 13) gives 110 mg (16% ) of the title compound. NMR of⁴H (400 MHz, CDCl1)₃) δ 1.05, 1.26, 3.05, 3.1, 3.2, 3.63.85, 3.86, 4.05, 6.95, 7.33, 7, 7, 8, 9, 9.6.

Example 44

1,2,3,3¹4,4 ', 4a, 6'-Octahydro-2', 4 ', 6'-trioxospiro [1, Hpyrazino [1,2-a] quinoline-5 (6ΈΓ), 5' (2'H) -pyrimidine-8carbonitrile monohydrochloride

<img file="PT1551849E_D0100.tif" />

* HCI

ΡΕ1551849

106

A suspension of 1,1-dimethylethyl 8-cyano-1,1'2,3 ', 4', 4a, 6'octahydro-2 ', 4', 6'-trioxospiro [3'-pyrazine [1 2-a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] -3-carboxylate of Example 29 (2.48 g, 5.83 mmol) is stirred in 4N HCl in dioxane (23 mL) for 1 h. The system is then purged with N 2 and the resulting solid is isolated by filtration. The white solid

<td>is crushed with</td><td>dioxane</td><td>and dry</td><td>(20 Torr, 110 ° C)</td><td>for</td>

<td>yield 1.77 g</td><td>(84%) of</td><td>salt of</td><td>hydrochloride. NMR</td><td>in ⁴H</td>

<td>(400 MHz, DMSO-ch</td><td>) δ 11.52</td><td> , 11,46,</td><td> 9,31, 9,10, 7,52,</td><td> 7, 47,</td>

<td> 7, 07, 4,33, 4, 14,</td><td> 3,38, 3,</td><td> 31, 3,27,</td><td> 3,15, 3,04, 2, 94,</td><td> 2, 78 .</td>

Example 45

2,3,4,4a-Tetrahydro-8-nitrospiro [1,2'-pyrazino [1,2-a] quinoline-5 (6H), 5 '(2'H) -pyrimidine-2', 4 ' , 6 '(1'H, 3'H) trione monohydrochloride

<img file="PT1551849E_D0101.tif" />

Similar to that described in Example 47, 47.996 mg (95%) of the title compound is obtained. (400 MHz, DMSO-kan) δ 11.59, 11.48, 9.20, 9.03, 8.00, 7.95, 7.09, 4.41, 4.25, 3.52, 3 , 44, 3.29, 3.11, 3.01, 2.86.

Example 4 6

Protocol for preparing various cyclic secondary amine analogs by parallel synthesis.

ΡΕ1551849

107

<img file="PT1551849E_D0102.tif" />

<img file="PT1551849E_D0103.tif" />

Ο

<img file="PT1551849E_D0104.tif" />

step 2 *

Step 1

An 8 mL screw cap vial is charged with 0.50 mmol of the desired amine followed by 0.7 mL (0.25 mmol) of a 0.35 M stock solution of the appropriate aldehyde in acetonitrile followed by 275 mg. (2.0 mmol) solid potassium carbonate. The vials are tightly closed and are then heated to 100 ° C in a whip block for 35 h. The cooled reactions are filtered into 20 mL vials containing 780 mg of Dowex P-SO3H resin (5.2 meq / g washed and dried). The vials are shaken at RT for 30-45 min, and then filtered into 20 mL pre-tare vials. The resin is washed with 85% CH 3 OH in H 2 O (4x1.5 mL). The resin is then eluted into predefined tare 20 mL vials using 2N pyridine in methanol. Both the wash and elution vials are concentrated in a Thermo Savant at 50 ° C overnight. The vials are weighed and the vials containing the product proceed to the next phase.

ΡΕ1551849

108

Step 2

To the vials containing the product from Step 1, an equivalent (as determined by the mmoles of the products from Step 1) of a 0.2 µl stock suspension is added.

M of barbituric acid in n-butanol. An additional 1 mL of n-butanol is added to each vial. The vials are tightly closed and heated to 100 ° C in a whip block for 18 h. The cooled reaction mixtures are diluted with approximately 4 mL of a 1: 1 acetonitrile / methanol solution. To each vial is added 1.5 g of strongly basic Dowex SBR resin (4.4 meq / g washed). The vials are shaken at RT for 2-3 h at RT and are filtered. The resin is washed with a small amount of 1: 1 acetonitrile / methanol solution. The resin is then eluted into predefined tare 20 mL vials using 10% TFA in acetonitrile solution (5x2 mL). The vials are concentrated in Thermo Savant at RT overnight to yield the final products. The weights of the final products are determined. The purity and mass identification of the end products is determined by LC / MS techniques.

Example 47

Protocol for preparing various analogs using commercially available benzaldehyde derivatives and cis-2,6-dimethylmorpholine.

ΡΕ1551849

109 &lt;rV

<img file="PT1551849E_D0105.tif" />

(8¼

<img file="PT1551849E_D0106.tif" />

Step 1

An 8 ml screw cap vial is charged with 0.5250 mmol of the desired aldehyde followed by

1.4 mL (0.5 mmol) of a 0.35 M stock solution of cis-2,6-dimethylmorpholine in acetonitrile followed by 275 mg (2.0 mmol) of solid potassium carbonate. The vials are tightly closed and are then heated to 100 ° C in a whip block for 35 h. The cooled reactions are filtered into 20 mL vials containing 780 mg of Dowex P-SO3H resin (5.2 meq / g washed and dried). The vials are shaken at RT for 30-45 min, and then filtered into 20 mL pre-tare vials. The resin is washed with 85% CH₃OH in H₂O (4x1.5 mL). The resin is then eluted into predetermined tare 20 mL vials using 2N pyridine in methanol. Both the wash and elution vials are concentrated in a Thermo Savant at 50 ° C overnight.

The vials are weighed and the vials containing the product proceed to the next phase.

ΡΕ1551849

110

Step 2

To the vials containing the Step 1 product is added one equivalent (as determined by the mmoles of the Step 1 products) of a 0.2 µm stock suspension.

M of barbituric acid in n-butanol. An additional 1 mL of n-butanol is added to each vial. The vials are tightly closed and heated to 100 ° C in a whip block for 18 h. The cooled reaction mixtures are diluted with approximately 4 mL of a 1: 1 acetonitrile / methanol solution. To each vial is added 1.5 g of strongly basic Dowex SBR resin (4.4 meq / g washed). The vials are shaken at RT for 2-3 h at RT and are filtered. The resin is washed with a small amount of 1: 1 acetonitrile / methanol solution. The resin is then eluted into predefined tare 20 mL vials using 10% TFA in acetonitrile solution (5 x 2 mL). The vials are concentrated in Thermo Savant at RT overnight to yield the final products. The weights of the final products are determined. The purity and mass identification of the end products is determined by LC / MS techniques.

Commercially Available Benzaldeldo derivatives include, but are not limited to: 2-Fluoroenzaldeldo;

2,3,4,5,6-Pentafluorobenzaldehyde; 2-Chloro-6-fluorobenzaldehyde; 2-Fluoro-6-chlorobenzaldeldo; 2,3-Difluorobenzaldehyde; 2,6-Difluorobenzaldehyde; 2,4-Difluorobenzaldehyde;

ΡΕ1551849

111

2.5-Difluoro-benzaldehyde; 2-Fluoro-5-nitrobenzaldehyde; 3-Cyano-4-dimethylamino-2-fluoro-benzaldehyde; 6- (Dimethylamino) -2-fluoro-3-formylbenzonitrile; 2-Fluoro-4,5-dimethoxybenzaldehyde; 3,4-Dimethoxy-6-fluoro-benzaldehyde, 2,3,6-Trifluorobenzaldehyde; 2,4,5-Trifluorobenzaldehyde; 2,4,6 Trifluorobenzaldehyde; 2,3,4-Trifluorobenzaldehyde; 2-Fluoro 3- (trifluoromethyl) benzaldehyde; 2-Fluoro-6- (trifluoromethyl) benzaldehyde; 2-Fluoro-4- (trifluoromethyl) benzaldehyde; 2Fluoro-5- (trifluoromethyl) benzaldehyde; 2-Fluoro-5-bromobenzaldehyde; 5-Bromo-2-fluorobenzaldehyde; 2-Fluoro-5-methoxybenzaldehyde; 6-Fluoro-m-anisaldehyde; 2,3,5-Trifluorobenzaldehyde; 2-Fluoro-4-bromo-benzaldehyde; 4-Bromo-2-fluoro-benzaldehyde; 2-Fluoro-4-chlorobenzaldehyde; 4-Chloro-2fluoro-benzaldehyde; 2,3,5,6-Tetrafluorobenzaldehyde; 2,3Difluoro-4- (trifluoromethyl) benzaldehyde; 2-Fluoro-4-methoxybenzaldehyde; 2-Fluoro-p-anisaldehyde; 4-Methoxy-2-fluorobenzaldehyde; 2-Fluorovaniline; 2-Fluoro-6-hydroxybenzaldehyde;

2-Fluoro-6-methoxybenzaldehyde; 3-Chloro-2,6-difluorobenzaldehyde; 2,6-Difluoro-3-methylbenzaldehyde; 3-Chloro-2-fluoro6- ​​(trifluoromethyl) benzaldehyde; 2-Chloro-6-fluoro-3-methylbenzaldehyde; 6-Chloro-2-fluoro-3-methylbenzaldehyde; 3-Chloro 3,6-difluorobenzaldehyde; 3-Chloro-2-fluoro-5- (trifluoromethyl) benzaldehyde; 3-Chloro-2-fluoro-benzaldehyde; 2,3-Difluoro-4-methylbenzaldehyde; 3-Fluoro-4-formylbenzenoboronic acid; 2-Fluoro-5-methylbenzaldehyde; 2,3-Difluoro-6-methoxybenzaldehyde; 3-Chloro-6-fluoro-2- (trifluoromethyl) benzaldehyde; 3-Fluoro-4-biphenylcarboxaldehyde; 2,3,4,5-Tetrafluorobenzaldehyde; 2-Fluoro-5-iodobenzaldehyde; 2,4-Dibromo-6-fluorobenzaldehyde; 3,5-Dibromo-2-fluorobenzaldehyde; 2ΡΕ1551849

112

Fluoro-4-methylbenzaldehyde; 4-Bromo-2,6-difluorobenzaldehyde; 4-Chloro-2,6-difluorobenzaldehyde; 4-Bromo-2,3,6-trifluorobenzaldehyde; 4-Chloro-2,3,6-trifluorobenzaldehyde; 5-Chloro2-fluorobenzaldehyde.

Example 48 rei- (2R, 4S, 4aS) -2,4-diethyl-1,2,4,4a-tetrahydro-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H ), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

<img file="PT1551849E_D0107.tif" />

Step 1: Preparation of 4-Benzyl-cis-2,6-diethylmorpholine

<img file="PT1551849E_D0108.tif" />

(DL + meso) -N-benzyl-1,1'iminbis-2-butanol (18.8 g, 74.6 mmol), described in J. Heterocycl. Chem; 20; 1983; 1681-1685, is cooled to 0 ° C and treated with 70% H₂ONLY₄(5 eq. W / w) with stirring. The mixture is sealed in a

ΡΕ1551849

113 pressure on glass and heated to 160 ° C for four days. The mixture is basified with 6.0 M NaOH until the solution is basic and extracted with MTBE (200 mL). The ether layer is washed with water and brine. The organic layer is dried over Na 2 SO₄ and filtered and the solvent is removed by rotary evaporation. The product is purified on a Biotage Flash 75L + using 7% MTBE in heptane as eluent. Product containing fractions are combined and the solvent is removed by rotary evaporation. The product is dried under vacuum resulting in a golden oil.

Step 2: Preparation of 2- (cis-2,6-Diethylmorpholin-4-yl) -5nitrobenzaldehyde

<img file="PT1551849E_D0109.tif" />

1-Chloroethyl chloroformate (2.18 mL, 20 mmol, Aldrich) is added to a solution of 4-benzyl-cis-2,6-dimethylmorpholine in CH₂C1₂ (45 mL) with stirring in an ice bath. The solution is allowed to warm to room temperature and stirred for 21 hours. The solvent is removed by rotary evaporation. The product is dissolved in CH 3 CN (40 mL) and treated with N, N-diisopropylethylamine (2.0 mL, 11.5 mmol, Aldrich). K 2 CO 3 (1.5 g, 11 mmol, Mallinkrodt) and 2-fluoro-5-nitrobenzaldehyde (1.9 g,

ΡΕ1551849

114

1.13 mmol, Oakwood) and the mixture is heated at 60 ° C for 21 hours. N, N, N'-trimethylethylenediamine (0.4 mL) is added and the reaction is stirred an additional 45 minutes at 60 ° C. The mixture is filtered and the solvent is removed by rotary evaporation. The residue is taken up in CH 2 Cl 2 (200 mL) and washed 2 x 1.0 M HCl, 2 x NaHCO 3 and 1 x brine (150 mL each). The organic layer is dried over Na 2 SO₄ and is filtered. The solvent is removed by rotary evaporation and the product is purified on a 40M + Biotage Flash using 15% EtOAc in heptane as eluent. Product containing fractions are combined and the solvent is removed by rotary evaporation. The product is dried under vacuum resulting in a yellow solid.

Step 3

2- (cis-2,6-Diethylmorpholin-4-yl) -5-nitrobenzaldehyde (1.58 g, 5.39 mmol) is combined with barbituric acid (767 mg, 5.39 mmol, Aldrich) in a MeOH vial (17 mL). The sealed reaction is stirred with heating at 80 ° C overnight. The solvent is removed by rotary evaporation and the product is purified on a 40M + Biotage Flash using 10% MTBE in CH 2 Cl 2 as eluent. The product is dried under vacuum and is recrystallized from CHsOH / heptane. The large crystals are triturated and dried under vacuum at 100 ° C resulting in 1.04 g (48% bright yellow crystalline solid. ⁴H (400 MHz, DMSO-cA) δ ppm 0.86, 0.97, 1.08, 1.24, 1.45, 1.58, 2.84, 3.03, 3.34, 3.60 , 3.97, 4.26, 7.01,

7, 84, 7, 98, 11,56, 11, 87.

ΡΕ1551849

115

Example 49 9- (2R, 4S, 4aS) -1,2,4,4a-tetrahydro-4-methyl-8-nitro-2- (trifluoromethyl) spiro [[1,4] oxazine [4,3- a] quinoline5 (6H), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

<img file="PT1551849E_D0110.tif" />

Step 1: Preparation of 3- (Benzylamino) -1,1,1-trifluoropan2-ol

<img file="PT1551849E_D0111.tif" />

Lithium triflate was suspended in CH₃CN and the mixture is cooled to -10 ° C. 2- (Trifluoromethyl) oxirane is added and the suspension is allowed to warm to RT with stirring. Benzylamine (6.33 mL, 58 mmol, Aldrich) is added in one portion and the resulting solution is stirred for 25 minutes. The reaction is quenched by pouring it into CH 2 Cl 2 (200 mL) and washing 3x with H₂O (200 mL). The organic layer is dried over Na₂ONLY₄ and is filtered. The product is purified on a Biotage Flash 75L using CH₂C1₂: pentane: triethylamine at 20:70:10 as eluent. The product is dried under vacuum at 100 ° C resulting in a white crystalline solid.

ΡΕ1551849

116

Step 2: Preparation of 4-Benzyl-2-methyl-6- (trifluoromethyl) morpholin-3-one

<img file="PT1551849E_D0112.tif" />

A solution of 3- (Benzylamino) -1,1,1-trifluoropropan-2-ol (2.68 g, 12.3 mmol) and triethylamine (1.7 mL, 12.1 mmol) in CH 2 Cl 2 is cooled to 0 ° C. The solution is treated dropwise with 2-bromopropanoyl chloride (1.2 mL, 12 mmol, Aldrich) and stirred for 5 minutes. The solution is diluted to 100 mL with CH 2 Cl 2 and sequentially washed with 1.0 M HCl, NaHCO 3 and brine (100 mL each). The organic layer is dried over Na₂ONLY₄ and is filtered. The solvent is removed by rotary evaporation yielding (N-benzyl-2-bromo-N- (3,3,3-trifluoro-2-hydroxypropyl) propanamide as a golden oil.) Dry NaH is suspended in inhibitor-free dry THF (25 ml) in a flame-dried flask and the mixture is cooled to 0 ° C with stirring in an ice bath. N-Benzyl-2-bromo-N- (3,3,3-trifluoro-2hydroxypropyl) propanamide (4.25 g, 12 mmol) is added as an inhibitor-free dry THF solution (25 mL) and the The resulting suspension is heated to RT. After stirring for one hour, the reaction is quenched with MeOH over an ice bath until no further gas evolution and

ΡΕ1551849

117 The mixture is poured into 1.0 M HCl (200 mL). The aqueous mixture is extracted into MTBE (200 mL) and the organic layer is washed with saturated NaHCO3 and brine (200 mL each). The organic layer is dried over Na₂ONLY₄ and is filtered, and the solvent is removed by rotary evaporation yielding an amber oil.

Step 3: Preparation of 4-Benzyl-cis-2-methyl-6- (trifluoromethyl) morpholine

<img file="PT1551849E_D0113.tif" />

LiAlH₄ (797 mg, 21 mmol, Aldrich) is suspended in inhibitor-free dry THF (15 mL) and treated with a solution of 4-benzyl-2-methyl-6- (trifluoromethyl) morpholin-3one (2.73 g, 10 mmol) in inhibitor-free dry THF (15 mL). The mixture is diluted with MTBE (25 mL) and sequentially quenched with H₂O (0.8 mL), 6.0M NaOH (0.8 mL) and H₂O (2.4 mL). The white slurry is filtered and the inorganic salts were washed with MTBE (200 mL). The solvent is removed by rotary evaporation and the product is purified in 3 portions on Biotage Flash 40M + cartridges using a 10 minute linear gradient of 3% -10% MTBE in heptane. The products are combined and dried under high vacuum to yield a colorless oil.

ΡΕ1551849

118

Step 4: Preparation of 2- [cis-2-Methyl-6- (trifluoromethyl) morpholin-4-yl] -5-nitrobenzaldehyde

<img file="PT1551849E_D0114.tif" />

1-Chloroethyl chloroformate (292 μΕ, 2.68 mmol, Aldrich) is added to a solution of 4-benzyl-cis-2-methyl6- (trifluoromethyl) morpholine (324 mg, 1.25 mmol) in CH 2 Cl 2 (6 mL) with stirring in an ice bath. The solution was allowed to warm to room temperature and stirred for 16 hours. The solvent is removed by rotary evaporation and the residue is dissolved in methanol (6 mL). This solution is heated at reflux for 4 hours and methanol is removed by rotary evaporation. 0 The residue is dissolved in CH 3 CN (5 ML) and treated with N, N-diisopropylethylamine (0.261 mL, 1.5 mmol, Aldrich). K 2 CO 3 (190 mg, 1.37 mmol, Mallinkrodt) and 2-fluoro-5-nitrobenzaldehyde (254 mg, 1.5 mmol) Oakwood) are added and the mixture is heated to 60 °.

C for 36 hours. N, N, N'-trimethylethylenediamine (2.3 mL) is added and the reaction is stirred for a further 2 hours at 60 ° C. The mixture is filtered and the solvent is removed by rotary evaporation. The residue is taken up in EtOAc (100 mL) and washed 2x with 1.0 M HCl, 1x saturated NaHCO 3 and

ΡΕ1551849

119 1x with saline (75 mL each). The organic layer is dried over Na₂ONLY₄ and is filtered. The solvent is removed by rotary evaporation and the product is purified over a 25M + Biotage Flash using 20% ​​EtOAc in heptane as eluent. Product containing fractions are combined and the solvent is removed by rotary evaporation. The product is dried under vacuum at 100 ° C yielding a yellow solid.

Step 5

2- [cis-2-methyl-6- (trifluoromethyl) morpholin-4-yl] 5-nitrobenzaldehyde (212 mg, 0.67 mol) is combined with barbituric acid (95 mg, 0.67 mmol, Aldrich) in a vial with MeOH (3 mL). The sealed reaction is heated to 80 ° C in a bump block for 21 hours and then the solvent is removed by rotary evaporation. The product is purified on a 25M + Biotage Flah using a 26 minute linear gradient of 2.5-10% MTBE in CH2 Cl2 eluent. The product is dried under vacuum and yielding a yellow-orange solid. 1.04 g (48% bright yellow crystalline solid.⁴H (400 MHz, DMSO-d₄) δ ppm 1.01, 2.93, 3.46, 3.66, 3.73, 4.04, 4.40, 4.48, 7, 10, 7, 88, 8, 01, 11.63 11.94.

Example 50 rei- (2R, 4S, 4aS) -1,2,4,4a-tetrahydro-4-methyl-8-nitro-2propilespiro [[1,4] oxazine [4,3-a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

ΡΕ1551849

120 ο₂ν

<img file="PT1551849E_D0115.tif" />

Step 1

Preparation of 2- [cis-2-Methyl-6-propylmorpholin-4-yl] -5nitrobenzaldehyde

A mixture of cis-allyl-4-benzyl-6-methylmorpholine (520 m, 2.25 mmol), trifluoroacetic acid (0.5 mL, Aldrich) and 5% palladium on carbon (54 mg, 10% w / w Aldrich) is sealed in a Parr bottle under 40 PSI of H₂ and is beat for 17 hours. The mixture is filtered and the solvent removed by rotary evaporation yielding cis-2-methyl-6-propylmorpholine trifluoroacetate as a light golden oil. The oil is taken up in CH 3 CN (8 mL) and treated with

N, N -diisopropylethylamine (1.2 mL, 6.9 mmol, Aldrich). 2-Fluoro-5-nitrobenzaldehyde (419 mg, 2.48 mmol,

Oakwood) and the mixture is stirred at 60 ° C for 21 hours.

N, N, N'-trimethylethylenediamine (1.0 mL) is added and the

ΡΕ1551849

121 The reaction is stirred for a further 30 minutes at 60 ° C after which time the solvent is removed by rotary evaporation. The residue is taken up in EtOAc (100 mL) and washed with 1.0 M HCl,

NaHCO₃ saturated saline solution (100 mL each). The organic layer is dried over Na₂ONLY₄ and filtered. The solvent is removed by rotary evaporation and the product is purified on a Biotage Flash 40M + using 20% ​​EtOAC in heptane as eluent. Product containing fractions are combined and the solvent is removed by rotary evaporation. The product is dried under vacuum at 60 ° C yielding a yellow oil.

Step 2

2- (cis-2-methyl-6-propylmorpholin-4-yl] -5-nitrobenzaldehyde (446 mg, 1.53 mol) is combined with barbituric acid (217 mg, 1.53 mmol, Aldrich) in a MeOH vial The sealed reaction is heated to 80 ° C in a tap block for 19 hours and then the solvent is removed by rotary evaporation.The product is purified 2x on a 75M Biotage Flah using 10% MTBE in eluent. CH₂C1₂. The product is dried under vacuum at 100 ° C yielding a yellow-orange solid. Y NMR (400 MHz, ACETONITRYL-D3) δ ppm 1.00, 1.47, 3.04, 3.34, 3.56, 3.95, 4, 14, 6.85, 7, 77.8 , 02, 9, 19.

Example 51 (2R, 4S, 4aS) -1,2,4,4a-tetrahydro-2,4-dimethyl-8-nitrospiro [[1,4] oxazine [4,3-a] quinoline-5 (6H ), 5 '(2'H) pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

ΡΕ1551849

122

<img file="PT1551849E_D0116.tif" />

Step 1: Preparation of (2S, 6S) -4-Benzyl-2,6-dimethylmorpholine (2S, 6S) -4-benzoyl-2,6-dimethylmorpholine (759 mg, 3.46 mmol) is dissolved in anhydrous THF ( 40 mL) and treated with LiAlH₄ (1.0 M in THF, 6.9 mL, Aldrich). The mixture is stirred with heating at 55 ° C for 18 hours. One more LiAlH equivalent is added₄ and the mixture is stirred with heating at 55 ° C for a further 6 hours. The reaction is quenched by adding water (0.4 mL) followed by 6.0 M NaOH (0.4 mL) followed by water (1.2 mL). White mud is diluted with Et₂O is filtered. Inorganic salts are washed with an additional portion of Et₂The combined ethereal washes are dried over Na₂ONLY₄. The product is purified over a 40M + Biotage Flash using 15% Et₂The in pentane as eluent. Product containing fractions are

ΡΕ1551849

123 combined and the solvent is removed by rotary evaporation. The product is dried under vacuum yielding a colorless oil.

Step 2: Preparation of 2- [2S, 6S) -2,6-Dimethylmorpholin-4-yl] 5-nitrobenzaldehyde

<img file="PT1551849E_D0117.tif" />

1-Chloroethyl chloroformate (292 μ] /, 2.68 mmol, Aldrich) is added to a solution of (2S, 6S) -4-benzyl-2,6-dimethylmorpholine (in CH₂C12 (6 mL) with stirring in an ice bath. The solution is allowed to warm to room temperature and stirred for 2.5 hours. The solvent is removed by rotary evaporation and the residue is dissolved in methanol (6 mL). This solution is heated at reflux for 2.5 hours and methanol is removed by rotary evaporation. The residue is dissolved and treated with N, N-diisopropylethylamine.

Aldrich). Add K₂CC&gt; 3 (190

Mallinkrodt) and 2-fluoro-5-nitrobenzaldehyde (230 mg, 1.36 mmol, Oakwood) and the mixture is heated at 60 ° C for 21 hours. The mixture is filtered and the solvent is removed by rotary evaporation. The residue is placed in CH₂C1₂ (100 mL) and washed sequentially with water (75 mL) and

<td>CH₃CN (5</td><td>mL)</td><td>and is</td>

<td>(244 μΕ,</td><td> 1,4</td><td>mmol,</td>

<td>mg, 1,</td><td> 37</td><td>mmol,</td>

saline (75 mL). The organic layer is purified on a Biotage.

ΡΕ1551849

124

Flash 25M + using 40% EtOAc in heptane as eluent. Product containing fractions are combined and the solvent is removed by rotary evaporation. The product is dried under vacuum resulting in a yellow syrup.

Step 3

2 - [[2S, 6S) -2,6-Dimethylmorpholin-4-yl] -5-nitrobenzaldehyde (298 mg, 1.12 mmol) is combined with barbituric acid (159 mg, 1.12 mmol, Aldrich) in a vial with MeOH (2 ML). The sealed reaction is heated to 60 ° C in a bump block for 21 hours and then the solvent is evaporated. The product is purified on a 40M + Biotage Flah using a 1: 1 mixture of EtOAc and toluene as eluent. The product is dried under vacuum resulting in a yellow-orange solid. NMR of⁴H (400 MHz, DMSO-d8) δ ppm 0.91, 1.24, 2.91, 3.56, 3.63, 3.83, 3.91, 3.95, 4.07, 4.14 -4.24, 6.85,

7,83, 7,98, 11,59, 11,89.

Example 52 rei- (2R, 4S, 4aS) -acetyl-10-fluoro-1,2,4,4a-tetrahydro-2,4-dimethylpiro [[1,4] oxazine [4,3-a] quinoline-5 (6ΈΓ), 5 '(2'H) pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

<img file="PT1551849E_D0118.tif" />

ΡΕ1551849

125

Step 1

<img file="PT1551849E_D0119.tif" />

<img file="PT1551849E_D0120.tif" />

NMR of ⁴H (400 MHz, DMSO-d₆) δ 11, 9, 11.5, 7.55, 7, 43,

4,00, 3,89, 3,75, 3,63, 3,54, 3,03, 2,93, 2,42, 1,11, 0,90.

Example 53 rei- (2R, 4S, 4aS) -8-acetyl-9,10-difluoro-1,2,4,4a-tetrahydro-2,4-dimethyl spiro [[1,4] oxazine [4,3-a ] quinoline5 (6ΈΓ), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

Step 1

<img file="PT1551849E_D0121.tif" />

<img file="PT1551849E_D0122.tif" />

ΡΕ1551849

126

Step 2

<img file="PT1551849E_D0123.tif" />

<img file="PT1551849E_D0124.tif" />

NMR of ⁴H 3.89, 3.74, (400 MHz, DMSO-cA) δ 11.9, 11.5,

3,64, 3,56, 3,07, 2,85, 2, 46, 1, 12,

7,28,

0,90.

4, 10,

Example 54 rei- (2R, 4S, 4aS) -10-fluoro-1,2,4,4a-tetrahydro-2,4-dimethyl8-nitrospiro [[1,4] oxazine [4,3-a] quinoline -5 (6ΈΓ), 5 '(2'H) pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

<img file="PT1551849E_D0125.tif" />

Nitric acid (2.8 dropwise to 1 ml solution) is slowly added 2,3-difluoro-benzaldehyde (2.0 g, Aldrich) in H₂ONLY₄ concentrate (14 ml) while cooling

ΡΕ1551849

127 at 0 ° C. The resulting mixture is warmed to room temperature and stirred for about 3 hours. The reaction is basified with NaHCO₃ saturated and extracted with MTBE (2x), washed with brine, dried over Na₃ONLY₄ and concentrated by rotovap to produce a brownish red oil. The resulting oil is purified in a 90 g Biotage cartridge.

Step 2

<img file="PT1551849E_D0126.tif" />

<img file="PT1551849E_D0127.tif" />

Nitro-difluoro-benzaldehyde (0.94 g) is dissolved in CH₃CN (10 ml) and Et₃N (1.23 ml). D-Methyl morpholine cis (0.78 ml) is added and the resulting mixture is refluxed for about 12 hours, cooled to room temperature and then to 0 ° C for about 3 hours. The resulting mixture is filtered and the solids are rinsed with cold IPA and then dried at 100 ° C.

Step 3

<img file="PT1551849E_D0128.tif" />

<img file="PT1551849E_D0129.tif" />

ΡΕ1551849

128 fluoro-nitro-aldehyde (0.5 g) and barbituric acid (0.243 g) are dissolved in IPA (8 ml) and refluxed for about 12 hours. The resulting mixture is cooled and stored at 0 ° C for about 2 hours. The mixture is filtered and the solids are rinsed with cold IPA and dried at 90 ° C for 2 days. NMR of⁴H (400 MHz, CDCl 3) δ 7.93, 7.78, 4.15, 3.97, 3.68, 3.10, 2.93, 1.13, 0.91.

Example 55

Oxadiazolyl Derivatives

Figure 55.1

<img file="PT1551849E_D0130.tif" />

<img file="PT1551849E_D0131.tif" />

Compounds synthesized according to scheme 1:

ΡΕ1551849

129

<img file="PT1551849E_D0132.tif" />

<img file="PT1551849E_D0133.tif" />

<img file="PT1551849E_D0134.tif" />

<img file="PT1551849E_D0135.tif" />

<img file="PT1551849E_D0136.tif" />

<img file="PT1551849E_D0137.tif" />

<img file="PT1551849E_D0138.tif" />

<img file="PT1551849E_D0139.tif" />

IR (diffuse reflectance) 1750, 1724, 1708, 1617, 1596, 1487, 1451, 1409, 1374, 1346, 1335, 1196, 826, 788, 754 cm^-1. MS (EI m / z (king intensity) 411 (M⁺, 99), 412 (21), 411 (99), 326 (46), 296 (23), 284 (20), 283 (51), 280 (28), 240 (25), 239 (20), 238 (22). HRMS (ESI) calc. for

C20H21N5O5 + H! 412.1621, found 412.1631.

ΡΕ1551849

130

Scheme 55.2:

<img file="PT1551849E_D0140.tif" />

Example 56 rei- (2R, 4S, 4aS) -1,2,4,4a-tetrahydro-2,4-dimethyl-8- (3methyl-1,2,4-oxadiazol-5-yl) spiro [[ 1,4] oxazine [4,3a] quinoline-5 (6H), 5 '(2'H) -pyrimidine] -2', 4 ', 6' (1'H, 3'H) trione (R = CH₃ in scheme 55.2)

Step 1

<img file="PT1551849E_D0141.tif" />

According to a procedure reported in Syn Comm 23 (22) 3149-3155 (1993), hydrogen peroxide (UHP, 3.76 g, 0.04 mol) is added to a nitrile solution (2.88 g, 0, 01 mol), potassium carbonate (0.14 g, 0.001 mol) in acetone (~ 10 mL) and water (~ 10 mL). The yellow solution is stirred overnight at room temperature. The mixture is concentrated in vacuo to remove organic solvent and

ΡΕ1551849

131 White mixture is diluted with ~ 10 mL of water. The mixture is stirred at room temperature for 1 h before filtration. The solid product is washed with water and vacuum dried at 60 ° C overnight to yield the desired amide product.

Step 2

<img file="PT1551849E_D0142.tif" />

A mixture of the amide acetal from step 1 (0.85 g, 2.77 mmol) and N, N-dimethylacetamide dimethyl acetal (~ 3mL) is heated to 120 ° C for 1.5 h. The dark solution is cooled to room temperature and concentrated in vacuo at 70 ° C to yield 1.33 g of dark liquid which is then treated with a 50% aqueous hydroxylamine solution (0.24 mL, 3.3 mL). mmol) at room temperature for 30 minutes. The mixture is diluted with 10% sodium bicarbonate solution and dichloromethane. The phases are separated and the organic phase is concentrated in vacuo to yield a brown oily product.

Step 3

<img file="PT1551849E_D0143.tif" />

ΡΕ1551849

132

A solution of crude exadiazole acetal from step 2, aqueous 2 N HCl (15 mL) and THF (15 mL) is heated to reflux for 30 minutes. The cooled solution is concentrated in vacuo to remove organic solvents. The aqueous phase is diluted with dichloromethane and aqueous sodium bicarbonate and the phases are separated. The organic phase is washed with water, dried (Na₂ONLY₄), concentrated in vacuo and chromatographed over silica gel (20 mL), eluting with 20% ethyl acetate / dichloromethane to afford an orange-yellow solid.

Step 4

<img file="PT1551849E_D0144.tif" />

<img file="PT1551849E_D0145.tif" />

A mixture of the aldehyde from step 3, (0.545 g, 1.81 mmol) and barbituric acid (0.234 g, 1.82 mmol) in methanol (~ 8 mL) is slowly heated to reflux under nitrogen for 18 h. The solution is cooled to room temperature and concentrated to yield a yellow slurry mixture which is suspended in an ether / hexane mixture. After cooling in an ice bath for 15 min., The mixture is filtered and the yellow solid is washed with ether to yield the title compound. IR (diffuse reflectance) 3212, 1757, 1727, 1709,

1705, 1614, 1498, 1429, 1397, 1392, 1348, 1338, 1286, 1248

ΡΕ1551849

133

<td>755 cnf¹.</td><td>MS</td><td>(Cl) m / z (intensity</td><td>King)</td><td> 412</td><td>(MH⁺, 99 )</td><td> , 414</td>

<td> (6), 413</td><td> (28:</td><td> ), 412 (99), 411 (4),</td><td> 410</td><td> (3) ,</td><td> 116 (3)</td><td> , 114</td>

<td> (17), 98</td><td> (3)</td><td> , 96 (10), 59 (4).</td><td>HRMS</td><td>(ESI</td><td>) callus.</td><td>for</td>

CscYiNsOs + Hi 412.1621, found 412.1630.

Example 57

Triazolyl derivatives

The following Scheme 57.1 illustrates an exemplary methodology for producing triazole derivatives.

Scheme 57.1

<img file="PT1551849E_D0146.tif" />

...............H

NaOMe, MeOH

O

Jl &lt;/ XA₀

MeOH.

<img file="PT1551849E_D0147.tif" />

Example 58

Amine / Het Derivatives¹ Couplings by Azoto

The following Scheme 58.1 illustrates an exemplary methodology for producing amine / het derivatives¹.

ΡΕ1551849

134

Figure 58.1

<img file="PT1551849E_D0148.tif" />

R ”

Buchwald Amination

R 'i

R *

<img file="PT1551849E_D0149.tif" />

CHO o

I

HN NH

N''V '

O

MeOH, Reflux

<img file="PT1551849E_D0150.tif" />

H⁺

Example 59

<img file="PT1551849E_D0151.tif" />

<img file="PT1551849E_D0152.tif" />

A solution of bromine compound (5.0 g, 12.25 mmol) and tetramethylethylenediamine (6.0 mL, 4.62 g, 39.83 mmol) in dry THF (75 mL) is cooled in an ice bath under nitrogen. A 3M solution of MeMgBr in ether (10.0 mL, 30.0 mmol) is added and the mixture is stirred for 15 min. The reaction is then cooled to -78 ° and a 1.7 M solution of t-BuLi in pentane (25.0 ml, 42.5 mmol) is added. The reaction is monitored by hplc to follow the disappearance of the bromine derivative. Additional tBuLi (7.0 ml, 11.9 mmol) is added after 1 h. 30 mins then dry DMF (10.0 mL, 9.44 g, 129.3 mmol) is added and the cooling bath is removed and the reaction is allowed to

ΡΕ1551849

135 heat up to temp. environment. Methanol (10 ml) is added and the solvent is evaporated. The red gel / gum residue is partitioned between ethyl acetate (200 ml) and water (150 ml). The pH is adjusted to 4-5 with conc. the aqueous is further extracted with ethyl acetate (2 x 250 ml) The organic extract is washed with water (2 x 150 ml), brine (100 ml), dried (MgSO4).₄), filtered and evaporated to yield an orange foam. Dissolution in ethyl acetate (25 mL) and filtration through silica gel (90 g) with ethyl acetate yields the aldehyde as an amber solid. NMR of⁴H (400 MHz, DMSO-d8) δ 0.93, 1.16, 2.87, 2.97, 3.48, 3.53, 3.61, 3.86, 4.26, 4.26, 7.03, 7.40, 7.61, 9.62, 11.48, 11.83.

Example 60 rei- (2R, 4S, 4aS) -1,2,4,4a-tetrahydro-2,4-dimethyl-8acetilespiro [[1,4] oxazine [4,3-a] quinoline-5 (6H ), 5 '(2Ή) pyrimidine] -2', 4 ', 6' (1'H, 3'H) -trione

<img file="PT1551849E_D0153.tif" />

Step 1: 2- (5-Bromo-2-fluorophenyl) -1,3-dioxolane

<img file="PT1551849E_D0154.tif" />

ΡΕ1551849

136

5-Bromo-2-fluorobenzaldehyde (14.6 mL, 0.123 mol,

Avocado) is combined with 4-methylbenzenesulfonic acid hydrate (2.34 g, 12.0 mmol, Aldrich), ethylene glycol (13.7 mL, 0.25 mol, Mallinkrodt) and toluene (75 mL). The mixture is heated to reflux overnight. The resulting solution is diluted with EtOAc (100 mL) and washed 2x with saturated NaHCO 3, 1x with water and 1x with brine (170 mL area). The organic layer is dried over Na₂ONLY₄ and filtered. The solvent is removed by rotary evaporation and the product is purified on a Biotage Flash 75L cartridge (800g silica) using 96: 3: 1 heptane: EtOAc: TEA as eluent.

Step 2: 5-Acetyl-2-fluorobenzaldehyde

<img file="PT1551849E_D0155.tif" />

2- (5-bromo-2-fluorophenyl) -1,3-dioxolane (5.2 g,

21.2 mmol) is taken up in dry, preservative-free THF (30 mL) and the solution is cooled to -40 ° C under N₂.

A 1.6 M solution of n-butyllithium in hexanes (13.2 mL, eq) is added via syringe and the solution is stirred for 25 minutes at -40 ° C. The lithium substrate is added dropwise. cannulated to a -40 ° C solution of N-methoxy-N-methylacetamide (2.36 mL, 22.2 mmol,

ΡΕ1551849

137

Aldrich) in 10 mL of preservative-free dry THF. The solution is heated to RT with stirring for 1 hour. The solution is poured into a vial containing 1.0 M HCl (100 mL) and heated to 65 ° C with vigorous stirring overnight. Upon cooling, the oily product is extracted into EtOAc (200 mL) and the organic layer is washed with water and brine (200 mL each). The organic layer is dried over Na 2 SO₄and is filtered. The solvent is removed by rotary evaporation and the product is purified on a Biotage Flash 75L cartridge (800 g silica) using 30% MTBE in heptane as eluent. Product containing fractions are combined and the solvent is removed by rotary evaporation. The product is dried under vacuum resulting in an oily yellow solid.

Step 3

<img file="PT1551849E_D0156.tif" />

The aldehyde from step 2 is combined with cis-2,6-dimethylmorpholine (1.18 mL, 9.75 mmol, TCI-US), K 2 CO 3 (2.15 g, 2.5 eq, Mallinkrodt) and CH 3 CN (12 mL) in a flask and the heterogeneous mixture is heated to 80 ° C overnight with vigorous stirring. After filtration, the solvent is removed by rotary evaporation. The orange oil is collected in

ΡΕ1551849

138

EtOAc (200 mL) and washed 2x with water and 1x with brine (175 mL each). The organic layer is dried over Na₂ONLY₄ and is filtered. The solvent is removed by rotary evaporation and the product is purified on a Biotage Flash 75L cartridge (800 g silica) using 30% MTBE in heptane as eluent. The product containing fractions are combined and the solvent is removed by rotary evaporation yielding 5-acetyl-2-cis- (2,6-dimethylmorpholin-4-yl) benzaldehyde as an orange-yellow oil. The aldehyde is collected in CH₃OH (10 mL) and combined with barbituric acid / 19 mg, 5.61 mmol,

Aldrich) in a sealed vial. The reaction is heated to

80 ° C in a bump block for 3.5 hours. The vial is cooled in the freezer to precipitate a mustard yellow solid. The solid is filtered off and washed with CH₃OH The solid was used as is for chiral resolution. NMR of⁴H (400 MHz, DMSO-cA) δ ppm 0.91, 1.14, 2.38, 2.88, 3.42, 3.51,

3,59, 3,81, 4,20, 6,91, 7,50, 7,69, 11,47, 11,79.

Example 61

Additional compounds of the invention may be produced by the methodology described herein as well as by methods known in the art. Examples of additional compounds of the invention include, but are not limited to the following compounds.

ΡΕ1551849

139

<img file="PT1551849E_D0157.tif" />

ΡΕ1551849

140

<img file="PT1551849E_D0158.tif" />

ΡΕ1551849

141

<img file="PT1551849E_D0159.tif" />

ΡΕ1551849

142

<img file="PT1551849E_D0160.tif" />

<img file="PT1551849E_D0161.tif" />

<img file="PT1551849E_D0162.tif" />

ΡΕ1551849

143

<img file="PT1551849E_D0163.tif" />

ΡΕ1551849

144

<img file="PT1551849E_D0164.tif" />

<img file="PT1551849E_D0165.tif" />

<img file="PT1551849E_D0166.tif" />

ΡΕ1551849

145

<img file="PT1551849E_D0167.tif" />

ΡΕ1551849

146

<img file="PT1551849E_D0168.tif" />

ΡΕ1551849

147

<img file="PT1551849E_D0169.tif" />

CF₃ ÕF_The ^3

ΡΕ1551849

148

<img file="PT1551849E_D0170.tif" />

ΡΕ1551849

149

<img file="PT1551849E_D0171.tif" />

ΡΕ1551849

150

<img file="PT1551849E_D0172.tif" />

<img file="PT1551849E_D0173.tif" />

<img file="PT1551849E_D0174.tif" />

<img file="PT1551849E_D0175.tif" />

^ N (CH3)₂

<img file="PT1551849E_D0176.tif" />

<img file="PT1551849E_D0177.tif" />

<img file="PT1551849E_D0178.tif" />

<img file="PT1551849E_D0179.tif" />

ΡΕ1551849

151

<img file="PT1551849E_D0180.tif" />

ΡΕ1551849

152

<img file="PT1551849E_D0181.tif" />

ΡΕ1551849

153

<img file="PT1551849E_D0182.tif" />

<img file="PT1551849E_D0183.tif" />

<img file="PT1551849E_D0184.tif" />

<img file="PT1551849E_D0185.tif" />

<img file="PT1551849E_D0186.tif" />

<img file="PT1551849E_D0187.tif" />

I

<img file="PT1551849E_D0188.tif" />

<img file="PT1551849E_D0189.tif" />

s

<img file="PT1551849E_D0190.tif" />

N ^ZS··'·

<img file="PT1551849E_D0191.tif" />

ΡΕ1551849

154

In vitro DNA Girase Assay

DNA gyrase is a bacterial topoisomerase that introduces negative super spirals into DNA. DNA gyrase assay measures the degree of enzymatic activity by quantifying the relative amounts of relaxed DNA vs. Supercoiled DNA on an EtBr-stained 0.8% agarose gel. The substrate is relaxed pBR322. The enzyme is E. DNA gyrase. coli which is purified from strains expressing each of the subunits individually (there are two: A and B and the holoenzyme consists of the A2B2 heterodimer). See, for example, Hallett et al. Cloning of tac-controlled DNA gyrase genes under control: overproduction of gyrase proteins A and B. Gene 93: 139142 (1990); Simon et al. In vitro biochemical complementation studies of gyrase subunits from different species. FEBS Lett 373: 88-92 (1995); and O'Dea et al. Mutations in the B subunit of Escherichiâ coli DNA gyrase that affect ATP-dependent reactions. J. Biol Chem 1996 271: 9723-9 (1996).

In the following record racemic mixtures are indicated with (q) - and enantiomerically enriched samples are indicated with (-) - or (+) -.

ΡΕ1551849

155

Made by

Example

Gyrase

IC50 Structure No. (μΜ)

<td> 1</td><td colspan="2">Η O w- °</td><td> 11, 79</td>

<td> 1</td><td>° Ί (±&gt; - s</td><td>Π r^N~ ^ NH V^O</td><td> 18, 96</td>

<td> 1</td><td>US k</td><td>Η the Af NH . °</td><td> 6,1</td>

<td></td><td colspan="2">(-) - enantiomer</td><td></td>

<td> 2</td><td colspan="2">Η O _ Oh no Ow (*) k ^ x®</td><td> 45,59</td>

<td> 3</td><td>% IJA (_±). k ^</td><td>Η the ---- NH V ° χθ</td><td> 23, 96</td>

ΡΕ1551849

156

Prepared by (continued)

Girase Example

IC50 Structure No. (μΜ)

<td rowspan="2"> 3</td><td rowspan="2">Xx</td><td colspan="3">Η the</td><td rowspan="2"> 46, 65</td>

<td>ζ'-ν, 7th</td><td colspan="2">NM T °</td>

<td></td><td colspan="3">mixture of isomers</td><td>cis and trans</td><td></td>

<td> 4</td><td></td><td></td><td></td><td>Π</td><td></td>

<td></td><td></td><td></td><td></td><td>NH</td><td></td>

<td></td><td>W</td><td rowspan="2"></td><td></td><td></td><td></td>

<td></td><td>OK (±) '</td><td colspan="2">.O Ύ</td><td> 70,2</td>

<td> 6</td><td></td><td> 0</td><td></td><td>Η η</td><td></td>

<td></td><td></td><td></td><td></td><td>NH</td><td></td>

<td></td><td></td><td rowspan="2">H l</td><td></td><td></td><td></td>

<td></td><td>V w-</td><td></td><td>v ° 7th</td><td> 13, 75</td>

<td> 18</td><td>O</td><td rowspan="2">O</td><td></td><td>H / 0 N-7</td><td> 8,4</td>

<td></td><td></td><td></td><td>Λ ^NH</td><td></td>

<td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>I</td><td></td><td colspan="2">Η V</td><td></td>

<td></td><td>U</td><td></td><td></td><td>A 0</td><td></td>

<td></td><td></td><td colspan="2"></td><td></td><td></td>

<td></td><td></td><td></td><td>L.</td><td>νθ</td><td></td>

<td></td><td> (±)-</td><td></td><td></td><td></td><td></td>

ΡΕ1551849

157

Prepared by (continued)

Girase Example

IC50 Structure No. (μΜ)

<td> 25</td><td>O₂n_ ^</td><td>tf O 4th ^nh| fh 4 '&lt; X.&gt; ko</td><td> 18,2</td>

<td></td><td colspan="2">(-) enantiomer</td><td></td>

<td> 25</td><td></td><td>yo , ΝΗ I read it.</td><td> 32</td>

<td></td><td>(t) -</td><td>fTT “</td><td></td>

<td></td><td></td><td>CU Ν · ^ ° .NH iT r&gt; &lt;C</td><td></td>

<td></td><td> (*)</td><td></td><td></td>

<td></td><td colspan="2">mixture</td><td></td>

<td> 43</td><td>P¹ k (±) -</td><td>y -JJ Y</td><td> 24</td>

Lisbon, September 20, 2010