Non-nucleoside reverse transcriptase inhibitors

专利摘要:
The present invention relates to compounds of formula (I) and pharmaceutically acceptable salts thereof, as inhibitors of HIV reverse transcriptase, wild type and several mutant strains. Formula I In Formula I above, R 2 is selected from the group consisting of H, F, Cl, (C 1-4 ) alkyl, (C 3-4 ) cycloalkyl and CF 3 , R 4 is H or Me, R 5 is H, Me or Et, R 4 and R 5 are not both Me, and if R 4 is Me, then R 5 cannot be Et, R 11 is Et, cyclopropyl, propyl, isopropyl or isobutyl, Q is selected from the group consisting of Formula (II), (III), (IV) and (V). Formula II Formula III Formula IV Formula V
公开号:KR20030017540A
申请号:KR1020027017173
申请日:2001-06-14
公开日:2003-03-03
发明作者:시모노브뤼노
申请人:베링거 잉겔하임 (캐나다) 리미티드；
IPC主号:

专利说明:

Non-nucleoside reverse transcriptase inhibitors
[2] Diseases known as AIDS are caused by strains known as human immunodeficiency virus (HIV), in particular HIV-1. In order for HIV to be replicated by the host cell, the information of the viral genome must be integrated into the DNA of the host cell. However, HIV is a retrovirus, meaning that its genetic information is in the form of RNA. Thus, the HIV replication cycle requires a step of transcription of the viral genome (RNA) into DNA, which in turn is the reverse of the normal chain. Transcription of viral RNA to DNA is accomplished by an enzyme that is suitably dubbed reverse transcriptase (RT). HIV virions contain a copy of RT along with viral RNA.
[3] Reverse transcriptases have a known enzymatic function, which acts as RNA dependent DNA polymerase, as ribonuclease and as DNA dependent DNA polymerase. When RT acts as an RNA dependent DNA polymerase, it transcribes a single standardized DNA copy of viral RNA. When RT acts as a ribonuclease, it destroys the original viral RNA and releases the DNA just generated from the original RNA. Finally, when RT acts as a DNA dependent DNA polymerase, the first DNA strand is used as a template to create a second complementary DNA strand. The two strands form a double standardized DNA, which is integrated into the genome of the host cell by another enzyme called integrase.
[4] Compounds that inhibit the enzyme function of HIV-1 reverse transcriptase will inhibit the replication of HIV-1 in infected cells. Such compounds include 3'-azido-3'-deoxythymidine (AZT), 2 ', 3'-dideoxynosine (ddI), 2', 3'-dideoxycytidine (ddC), d4T, 3TC It is useful for the prevention or treatment of HIV-1 infection in human subjects, as indicated by known RT inhibitors such as nevirapine, delavirdin, epavirens and abacavir, and therefore, the main agent is to be used for the treatment of AIDS. Already approved
[5] With any antiviral therapy, the use of RT inhibitors in the treatment of AIDS ultimately leads to viruses that are less sensitive to a given drug. Resistance to these agents (reduced sensitivity) is the result of mutations that occur in the reverse transcriptase fragment of the Paul gene. Several mutant strains of HIV are characterized and resistance to known therapeutic agents is due to mutations in the RT gene. Some of the most clinically observed mutants include the Y181C mutant, wherein tyrosine (Y) at codon 181 has been mutated to a cysteine (C) residue, and K103N (wherein lysine (K) at position 103 is asparagine). (N)). Other mutants that show increased frequency during treatment with known antiviral agents include single mutants V106A, G190A, T188C, and P236L, and double mutants include K103N / Y181C, K103N / P225H, K103N / V108I, and K103N / L100I is included.
[6] As prevention and treatment of HIV infection with antiviral agents, the incidence of new resistant strains is expected to increase. Thus, there is a continuing need for new inhibitors of RT that have different patterns of effectiveness against various mutants.
[7] Compounds having a tricyclic structure that are inhibitors of HIV-1 are described in US Pat. No. 5,366,972. Other inhibitors of HIV-1 reverse transcriptase are described in Hargrave at el., J. Med Chem., 34, 2231 (1991).
[8] U.S. Patent 5,705,499 discloses 8-arylalkyl- and 8-arylheteroalkyl-5,11-dihydro-6H-dipyrido [3,2-B: 2 ', 3'-E] [1 as inhibitors of RT. , 4] diazepine has been proposed. The compounds exemplified have been shown to have some activity against wild type and mutant HIV-1 RT, in particular Y181C and less effective but other single mutants such as K103N.
[9] Specifically, the compounds of the present invention are effective at inhibiting not only Y181C and K103N but also a wide range of other single mutants and certain commonly found double mutants such as K103N / Y181C and K103N / P225H.
[10] Summary of the Invention
[11] The present invention provides novel compounds that are potent inhibitors of single and double mutant strains of HIV-1 RT to reduce the difficulties and disadvantages of the prior art.
[12] In a first aspect, the invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof.
[13]
[14] In Formula I above,
[15] R ² is selected from the group consisting of H, F, Cl, C _1-4 alkyl, C _3-4 cycloalkyl and CF ₃ ,
[16] R ⁴ is H or Me,
[17] R ⁵ is H, Me or Et, R ⁴ and R ⁵ are not both Me, and if R ⁴ is Me, then R ⁵ cannot be Et,
[18] R ¹¹ is Me, Et, cyclopropyl, propyl, isopropyl or cyclobutyl,
[19] Q is selected from the group consisting of Formula (II), (III), (IV) and (V).
[20]
[21]
[22]
[23]
[24] In a second aspect, the invention provides an HIV replication inhibitor of formula (I) or a pharmaceutically acceptable salt thereof.
[25] In a third aspect, the present invention provides a reverse transcriptase inhibitor of HIV of formula (I) or a pharmaceutically acceptable salt thereof.
[26] In a fourth aspect, the invention provides a method of treating or preventing HIV infection comprising administering to a patient an HIV inhibitory amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.
[27] In a fifth aspect, the present invention provides a pharmaceutical composition for treating or preventing HIV infection, comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.
[28] In a sixth aspect, the invention provides a process for the preparation of a compound of formula (I) or a pharmaceutically acceptable salt thereof.
[1] The present invention relates to novel compounds and pharmaceutically acceptable salts thereof, to their use alone or in combination with other therapeutic agents in the treatment or prevention of HIV infection, and to pharmaceutical compositions comprising the compounds.
[29] Justice
[30] As used herein, the term “C _1-4 alkyl” is intended to mean straight or branched chain alkyl radicals of 1 to 4 carbon atoms, and methyl, ethyl, propyl, isopropyl, butyl, secondary butyl and tertiary butyl. Include.
[31] As used herein, the term “C _3-4 cycloalkyl” is intended to mean a saturated cyclic hydrocarbon radical having 3 to 4 carbon atoms and includes cyclopropyl and cyclobutyl.
[32] Detailed Description of the Preferred Embodiments
[33] According to a preferred embodiment, the compounds of the invention are defined according to formula (I), wherein R ² is preferably Cl, F or H. More preferably, R ² is Cl or H. Most preferably, R ² is H.
[34] According to a preferred embodiment, the compounds of the invention are defined according to formula (I) wherein R ⁴ is preferably H.
[35] According to another embodiment, the compounds of the invention are preferably defined according to formula (I), wherein R ⁵ is Me.
[36] Preferably, the compounds of the present invention are defined according to formula (I), wherein R ¹¹ is Et or cyclopropyl. More preferably, R ¹¹ is Et.
[37] According to a preferred embodiment, the compound of the invention is preferably Q It is defined according to formula (I) selected from the group consisting of:
[38] More preferably, Q is to be.
[39] In general, preferred embodiments of the present invention , And It includes a compound selected from the group consisting of.
[40] The compounds of the present invention are not only effective inhibitors of wild type reverse transcriptases but also inhibit, for example, the single mutant enzymes Y181C, K103N, V106A, G190A, Y188C and P236L. The compound also inhibits double mutant enzymes K103N / Y181C, K103N / P225H, K103N / V108I and K103N / L100I.
[41] Compounds of formula (I) have inhibitory activity against HIV-1 reverse transcriptase. It is useful for the treatment of disorders associated with AIDS, ARC and HIV-1 infection when administered in a suitable dosage form. Accordingly, another aspect of the present invention is a method of treating HIV-1 infection comprising administering to a person infected with HIV-1 a therapeutically effective amount of the novel compound of formula (I) as described above. Whether for therapeutic or prophylactic purposes, the compound can also be used to prevent prenatal delivery before and after delivery from mother to baby by administering to the mother before birth.
[42] The compounds of formula (I) may be administered in single or divided doses by the oral or parenteral route. Suitable oral administration for the compound of formula (I) will range from about 0.5 mg to 1 g per day. Preferred oral administration to a compound of formula (I) will range from about 100 to 800 mg per day for a 70 kg body weight patient. In parenteral formulations, suitable dosage units may contain 0.1 to 250 mg, preferably 1 to 200 mg of the compound. However, it is to be understood that dosage administration varies from patient to patient and that administration to any particular patient depends on the clinician's judgment using the patient's response to the drug as well as the patient's body and condition as a criterion to fix the appropriate administration.
[43] When the compound of the present invention is to be administered by oral administration, it may be administered as a drug in the form of a pharmaceutical preparation, which contains the compound of the present invention together with a compatible pharmaceutical carrier material. Such carrier material may be an inert organic or inorganic carrier material suitable for oral administration. Examples of such carrier materials include water, gelatin, talc, starch, magnesium stearate, gum arabic, vegetable oil, polyalkylene-glycol, petroleum jelly and the like.
[44] Pharmaceutical formulations may be prepared by conventional methods and the processed dosage forms may be solid dosage forms, eg tablets, dragees, capsules or the like or liquid dosage forms such as solvents, suspensions, emulsions and the like. . Pharmaceutical formulations can be processed by conventional pharmaceutical operations such as sterilization. In addition, the pharmaceutical preparations may contain conventional adjuvants such as preservatives, stabilizers, emulsifiers, flavor improvers, wetting agents, buffers, salts to improve osmotic pressure. Solid carrier materials that can be used include, for example, starch, lactose, mannitol, methyl cellulose, microcrystalline cellulose, talc, silica, dibasic calcium phosphate and high molecular weight polymers such as polyethylene glycol.
[45] For parenteral use, the compounds of formula (I) are suspensions or emulsions, antioxidants, preservatives, buffers or blood and solution isotonic in aqueous or non-aqueous solutions, mixtures of pharmaceutically acceptable oils or liquids which may contain bacteriostatic agents It may be administered in other solutes, thickeners, suspensions or other pharmaceutically acceptable additives. Additives of this type are, for example, tartrate, citrate and acetate buffers, ethanol, propylene glycol, polyethylene glycols, complexing agents (e.g. EDTA), antioxidants (e.g., for viscosity control and polyethylene derivatives of sorbitol anhydride). Sodium sodium sulfate, sodium metasulphate and ascorbic acid) and high molecular weight polymers such as liquid polyethylene oxide.
[46] The compounds of the present invention may also be administered as solutions for application to the nose and may contain suitable buffers, tonicity aids, microbial preservatives, antioxidants and viscosity increasing agents in addition to the compounds of the present invention in aqueous vehicles. Examples of agents used for increasing the viscosity include polyvinyl alcohol, cellulose derivatives, polyvinylpyrrolidone, polysorbates or glycerin. The microbial preservative to be added may include benzalkonium chloride, thimerosal, chlorobutanol or phenylethyl alcohol.
[47] In addition, the compounds provided by the present invention may be administered as suppositories.
[48] The compounds of the present invention can be prepared using the techniques of synthetic organic chemists. Exemplary schemes are shown in Schemes 1-6.
[49]
[50]
[51] Continuation of formula (2) is described in J.M. Klunder et al .; J. Med. Chem. 1998, 41, 2960-71, and C.L. Cywin et al .; J. Med. Chem. 1998, 41, 2972-84.
[52]
[53]
[54]
[55]
[56] As previously described, the compounds provided by the present invention inhibit the enzymatic activity of HIV-1 RT. As described below, based on the testing of these compounds, the compounds of the present invention are known to inhibit the RNA dependent DNA polymerase activity of HIV-1 RT. The compound is also known to inhibit the DNA dependent DNA polymerase activity of HIV-1 RT (data not shown). Using the reverse transcriptase (RT) assay described below, compounds can be tested for their ability to inhibit RNA dependent DNA polymerase activity of HIV-1 RT. The specific special compounds described in the examples appearing below are thus tested. The results of this test are shown as IC ₅₀ (nM) in Table 2 and as EC ₅₀ (nM) in Table 3.
[57] The invention is further elucidated with the following non-limiting examples. All reactions are carried out under nitrogen or argon atmosphere. The temperature is given in ° C. Solution percentages or ratios are expressed in volume-to-volume relationship, unless otherwise noted.
[58] As used herein, abbreviations or symbols include the following:
[59] DEAD: diethyl azodicarboxylate,
[60] DIAD: diisopropyl azodicarboxylate,
[61] DIEA: diisopropylethylamine,
[62] DMAP: 4- (dimethylamino) pyridine,
[63] DMSO: dimethyl sulfoxide,
[64] DMF: dimethylformamide,
[65] ES MS: Electron Spray Mass Spectrometer,
[66] Et: ethyl,
[67] EtOAc: ethyl acetate,
[68] Et ₂ O: diethyl ether,
[69] HPLC: high performance liquid chromatography,
[70] iPr: isopropyl,
[71] Me: methyl,
[72] MeOH: methanol,
[73] MeCN: acetonitrile,
[74] NBS: N-bromosuccinimide,
[75] Ph: phenyl,
[76] TBE: Tris-Borate-EDTA,
[77] TBTU: 2- (1H-benzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium tetrafluoroborate,
[78] TFA: trifluoroacetic acid,
[79] THF: tetrahydrofuran,
[80] MS (ES): Electrospray Mass Spectrometer,
[81] MS (FAB) or FAB / MS: high speed atomic impact mass spectrometer,
[82] HRMS: high resolution mass spectrometer,
[83] PFU: plaque forming unit,
[84] DEPC: diethyl pyrocarbonate,
[85] DMSO: dimethyl sulfoxide,
[86] DTT: dityrotreitol,
[87] EDTA: ethylenediaminetetraacetate,
[88] UMP: Uridine 5'-monophosphate,
[89] UTP: Uridine 5'-triphosphate.
[90] synthesis
[91] The following examples illustrate the preparation of the compounds of the present invention.
[92] Example I (No. 24, Table 1)
[93] 5,11-dihydro-11-ethyl-5-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[94] (a) 2- (ethylamino) -3-nitropyridine
[95] To a solution of 2-chloro-3-nitropyridine (51 g, 325 mmol) in THF (650 mL) was added a 2 M solution of ethylamine in THF (365 mL, 731 mmol). The reaction is stirred overnight at room temperature. The reaction mixture is poured into water (˜1.5 L) and the solid obtained is filtered and dried under reduced pressure to give the title compound (52 g).
[96] (b) 3-amino-2- (ethylamino) pyridine
[97] A solution of 2- (ethylamino) -3-nitropyridine (52 g) in MeOH (600 mL) is stirred overnight at room temperature under hydrogen (1 atm) in the presence of 20% Pd (OH) _{2 /} C (10.4 g). The catalyst is removed by filtration through diatomaceous earth. The filtrate is concentrated under reduced pressure to give the title compound (39 g, 88% yield through steps (a) and (b)) as a black solid.
[98] (c) 2-chloro-N- {2- (ethylamino) -3-pyridinyl} -5-bromo-3-pyridinecarboxamide
[99] To a cold solution of 3-amino-2- (ethylamino) pyridine (30.6 g, 223 mmol) in MeCN (740 mL) was added solid NaHCO ₃ (56.3 g, 669 mmol). After 5 minutes, prepared from crude 5-bromo-2-chloro-3-pyridinecarbonyl chloride [5-bromo-2-hydroxy-3-pyridinecarboxylic acid and SOCl ₂ {except for aqueous workup) And TW Gero et al., Synth. Commun. 1989, 19, 553-559, which is incorporated herein by reference)] (1 equivalent, 223 mmol). After 2 hours, the reaction mixture is poured into ice / H ₂ O (1.5 L) and the solid obtained is filtered and rinsed with H ₂ O followed by hexane. After drying under reduced pressure overnight, the title compound (54.9 g, yield 69%) is obtained as a black solid.
[100] (d) 8-bromo-5,11-dihydro-11-ethyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[101] THF in 2-chloro-N- {2- (ethylamino) -3-pyridinyl} -5-bromo-3-pyridinecarboxamide (54.9 g, 154.4 mmol) in pyridine (308 mL) at 50 ° C. 335 mL, 335 mmol) of NaHMDS (sodium hexamethyldisilazide) 1M solution is added dropwise. After 10 minutes, the reaction is cooled to room temperature and then poured into ice water (2 L). The solid obtained is filtered and rinsed with water followed by hexane. The solid is dried under reduced pressure to afford the title compound (36 g, 75% yield) as a dark green solid.
[102] (e) 8-bromo-5,11-dihydro-11-ethyl-5-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine- 6-on
[103] 8-Bromo-5,11-dihydro-11-ethyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6 in DMF (380 mL) To a warm (36.7 g, 115 mmol) solution is added NaH (3.5 g, 138 mmol) and the mixture is heated at 50 ° C. for 30 minutes. The reaction mixture is cooled to room temperature and treated with MeI (14.3 mL, 230 mmol). After 1.5 hours, the reaction mixture is poured into ice water. The solid is filtered off, washed with water then hexanes to give the title compound (37.9 g, 99% yield) as a dark gray solid after drying.
[104] (f) 5,11-dihydro-11-ethyl-5-methyl-8- (2-propenyl) -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4 Diazepine-6-one
[105] Allyltributyltin (30.7 mL, 99.0 mmol) and Pd (Ph ₃ P) ₄ (5.20 g, 4.50 mmol) were added to 8-bromo-5,11-dihydro-11-ethyl in DMF (450 mL) at room temperature. To a degassed solution (passed through N ₂ for 30 minutes) of -5-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one Add. The mixture is stirred at 90 ° C. for 1.5 h, then cooled to rt and concentrated under reduced pressure. The residue is purified by flash chromatography (hexane: EtOAc, 8: 2 to 7: 3) to afford the title compound (22.19 g, yield 84%).
[106] (g) 5,11-dihydro-11-ethyl-8- (2-hydroxyethyl) -5-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1, 4] diazepine-6-one
[107] The ozonated oxygen stream was subjected to 5,11-dihydro-11-ethyl-5-methyl-8- (2-propenyl) -6H-dipyrido [3] in CH ₂ Cl ₂ (150 mL) and MeOH (150 mL). , 2-b: 2 ', 3'-e] [1,4] diazepine-6-one (22.19 g, 75.4 mmol) was bubbled through a cold (-78 ° C) solution for 2.5 hours. The N ₂ stream is then bubbled through the solution for 15 minutes and then solid NaBH ₄ (4.99 g, 132 mmol) is added to the solution. The reaction mixture is allowed to warm up to room temperature. After 1 h, aqueous saturated NH ₄ Cl (200 mL) is added and the mixture is stirred at rt for 2 h. The organic solvent is removed under reduced pressure. Water (300 mL) and CHCl ₃ (300 mL) were added to the residue. The phases are separated and the aqueous layer is extracted with CHCl ₃ (3 × 300 mL). The combined organic layers are dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (EtOAc: CHCl ₃ , 4: 1) to afford the title compound (16.1 g, 72% yield) as a white solid.
[108] (h) 5,11-dihydro-11-ethyl-5-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3' -e] [1,4] diazepine-6-one
[109] Diethyl azodicarboxylate (DEAD) (12.8 mL, 81.0 mmol) was added to 5,11-dihydro-11-ethyl-8- (2-hydroxyethyl) -5-methyl- in THF (270 mL) at room temperature. 6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (16.1 g, 54.0 mmol), 4-hydroxyquinoline (11.6 g, 81.0 mmol) And Ph ₃ P (21.3 g, 81.0 mmol) drop wise. The mixture is stirred at rt for 1 h and then concentrated under reduced pressure. The residue was purified by flash chromatography (EtOAc: MeOH; 95: 5) to give the title compound (17.7 g, yield 77%) as a white solid: MS (ESI) m / z 426 (MH) ⁺
[110] Example II (No. 2, Table 1)
[111] 2-chloro-5,11-dihydro-11-ethyl-5-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3 '-e] [1.4] diazepine-6-on
[112] (a) 6-chloro-2- (ethylamino) -3-nitropyridine
[113] EtNH ₂ (49.8 g, 1.10 mol) ice-cold solution in toluene (200 mL) was added to 2,6-dichloro-3-nitropyridine (100.0 g, 0.52 mol) ice-cold solution in toluene (225 mL) over 15 minutes. . The mixture is stirred at 0 ° C. for 45 minutes. Water (500 mL) and EtOAc (500 mL) are added and the phases are separated. The organic layer is washed successively with water (200 mL) and brine (200 mL), dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residual solid is recrystallized from MeOH to give the title compound (83.7 g, 80% yield) as a yellow needle.
[114] (b) 3-amino-6-chloro-2- (ethylamino) pyridine
[115] A solution of SnCl ₂ H ₂ O (616.3 g, 2.73 mol) in aqueous 12N HCl (500 mL) was dissolved in 6-chloro-2- (ethylamino) -3-nitropyridine (169.5 g, 0.84 mol) is added rapidly to the solution. After 20 minutes, the mixture is cooled to 0 ° C. and water (250 mL) is added. A small amount of solid NaOH (240 g) is then added. The suspension obtained is filtered to remove tin salt. The filtrate is diluted with water (3.5 L) and the solution is made basic by addition of aqueous 10N NaOH and then extracted with EtOAc (3 x 1.7 L). The combined organic layers are washed with brine (1 L), dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (hexane: EtOAc, 3: 2) to afford the title compound (89.9 g, yield 62%) as a brown solid: MS (ESI) m / z 172/174 (MH) ⁺
[116] (c) 5-bromo-2-chloro-N- {2- (ethylamino) -6-chloro-3-pyridinyl} -3-pyridinecarboxamide
[117] A solution of 5-bromo-2-chloro-3-pyridinecarbonyl chloride (30.0 g, 97.0 mmol) in MeCN (100 mL) in MeCN (180 mL) containing solid NaHCO ₃ (14.2 g, 169 mmol) at room temperature. To a 3-amino-6-chloro-2- (ethylamino) pyridine (16.6 g, 97.0 mmol) solution is added via cannula. The mixture is stirred at rt for 1 h. Water (200 mL) is added and the mixture is stirred for 10 minutes. The suspension obtained is filtered. The solid is washed with Et ₂ O (50 mL) and concentrated from pyridine solution (3 × 50 mL) to give the title compound (28.4 g, 75% yield).
[118] (d) 8-bromo-2-chloro-5,11-dihydro-11-ethyl-6H-dihydro [3,2-b: 2 ', 3'-e] [1,4] diazepine- 6-on
[119] NaHMDS 1M solution in THF (167.5 mL, 167.5 mmol) was heated to 50 ° C. in pyridine (146 mL) 5-bromo-2-chloro-N- {2- (ethylbromo) -6-chloro-3- To pyridinyl} -3-pyridinecarboxamide (28.4 g, 72.8 mmol) solution. The reaction mixture is stirred at 50 ° C. for 1.5 h. The mixture is then introduced into a mixture of water and ice (1 L) and after 1 hour the resulting suspension is filtered. The solid is washed with water and dried under reduced pressure to give the title compound (23.4 g, 91% yield).
[120] (e) 8-bromo-2-chloro-5,11-dihydro-11-ethyl-5-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4 Diazepine-6-one
[121] Solid NaH (60% oil dispersion, 3,46 g, 86.1 mmol) was added to 8-bromo-2-chloro-5,11-dihydro-11-ethyl-6H in DMF (220 mL) at 50 ° C. over 30 minutes. Add to dihydro [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (23.4 g, 66.3 mmol) solution. The mixture is stirred at 50 ° C. for 1 hour and then cooled to room temperature. The mixture is poured into water (1 L) and the suspension obtained is filtered. The solid is washed successively with water and hexanes and then dried under reduced pressure to afford the title compound (23.0 g, 94% yield).
[122] (f) 2-chloro-5,11-dihydro-11-ethyl-5-methyl-8- (2-propenyl) -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[123] Allyltributyltin (21.3 mL, 68.7 mmol) and Pd (Ph ₃ P) ₄ (3.61 g, 3.12 mmol) were added to 8-bromo-2-chloro-5,11-dihydro-11 in DMF (312 mL). -Ethyl-5-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (23.0 g, 62.5 mmol) degassing solution (through solution Pass through N ₂ for 30 minutes). The mixture is heated at 90 ° C. for 2 hours. The mixture is concentrated under reduced pressure. The residue is purified by flash chromatography (hexane: EtOAc, 7: 3) to give the title compound (13.4 g, yield 65%).
[124] (g) 2-chloro-5,11-dihydro-11-ethyl-8- (2-hydroethyl) -5-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[125] Ozonated oxygen was removed in 2-chloro-5,11-dihydro-11-ethyl-5-methyl-8- (2-pro) in MeOH (102 mL) and CH ₂ Cl ₂ (102 mL) until the alkene disappeared completely. Phenyl) -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (13.4 g, 40.7 mmol) introduced into a cold (-78 ° C) solution do. Nitrogen is bubbled through the solution to remove excess O ₃ . Then, a small amount of solid NaBH ₄ (2.69 g, 71.1 mmol) is added and the mixture is slowly warmed to room temperature. After 1 h, aqueous saturated NH ₄ Cl (150 mL) is added and the mixture is stirred for 20 minutes. The organic solvent is removed under reduced pressure. Water (100 mL) is added to the aqueous solution. The solution is extracted with CHCl ₃ (3 × 200 mL). The combined organic layers are dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (EtOAc: CHCl ₃ , 4: 1) to afford the title compound (10.4 g, 77% yield).
[126] (h) 2-chloro-5,11-dihydro-11-ethyl-5-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[127] Diethyl azodicarboxylate (DEAD) (4.26 mL, 27.0 mmol) was added to 2-chloro-5,11-dihydro-11-ethyl-8- (2-hydroethyl) -5 in THF (90 mL) at room temperature. -Methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (6.00 g, 18.0 mmol), 4-hydroxyquinoline (3.92 g, 27.0 mmol) and Ph ₃ P (7.09 g, 27.0 mmol) dropwise. The mixture is stirred at rt for 1 h and then concentrated under reduced pressure. The residue was purified by flash chromatography (EtOAc: MeOH, 95: 5) to give the title compound (6.22 g, yield 75%) as a white solid: MS (ESI) m / z 460/462.
[128] Example III (No. 6, Table 1)
[129] 2-chloro-11-cyclopropyl-5,11-dihydro-5-methyl-8- {2- (4-quinolinyloxy) -ethyl} -6H-dipyrido [3,2-b: 2 ' , 3'-e] [1,4] diazepine-6-one
[130] (a) 6-chloro-2- (cyclopropylamino) -3-nitropyridine
[131] A solution of cyclopropylamine (1.25 g, 22.0 mmol) in toluene (11 mL) was added to an ice cold solution of 2,6-dichloro-3-nitropyridine (2.00 g, 10.4 mmol) in toluene (10 mL) over 10 minutes. do. The mixture is stirred at 0 ° C. for 1 hour and at room temperature for 2 hours. Water (50 mL) is added to the mixture and the phases are separated. The organic layer is washed with brine (25 mL), dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (hexane: EtOAc, 3: 2) to afford the title compound (1.97 g, yield 89%) as a yellow solid.
[132] (b) 2-chloro-11-cyclopropyl-5,11-dihydro-8- (2-hydroxyethyl) -5-methyl-6H-dipyrido [3,2-b: 2 ', 3'- e] [1,4] diazepine-6-one
[133] The title compound is prepared from 6-chloro-2- (cyclopropylamino) -3-nitropyridine using a method similar to that described for 11-ethyl homolog in Example II.
[134] (c) 2-chloro-11-cyclopropyl-5,11-dihydro-5-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[135] Diethyl azodicarboxylate (DEAD) (85.6 μl, 0.54 mmol) was added to 2-chloro-11-cyclopropyl-5,11-dihydro-8- (2-hydroxyethyl) in THF (1.8 mL) at room temperature. -5-Methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one (125 mg, 0.36 mmol), 4-hydroxyquinoline (78.9 mg , 0.54 mmol) and Ph ₃ P (143 mg, 0.54 mmol) dropwise. The mixture is stirred at rt for 3 h and then concentrated under reduced pressure. The residue is partially purified by flash chromatography (EtOAc: MeOH, 95: 5). The solid was further purified by reverse phase HPLC (CombiPrep ADS-AQ 50 × 20 mm, 5μ, 120 A, 5-100% MeCN + 0.10% TFA / water + 0.10% TFA, 25min) to give the trifluoro of the title compound as a white solid. Acetic acid salt (17.7 g, yield 77%) is obtained: MS (ESI) m / z 472/474 (MH) ⁺ .
[136] Example IV (No. 4, Table 1)
[137] 5,11-dihydro-11-ethyl-2-fluoro-5-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[138] (a) 2,6-difluoro-3-nitropyridine
[139] 2,6-difluoropyridine (200 g, 1.74 mol) is added dropwise to a mixture of concentrated sulfuric acid (600 mL) and fuming nitric acid (90%, 400 mL) in an ice bath (keeping the internal temperature at 5-10 ° C). The resulting mixture is stirred overnight at room temperature. The mixture is slowly poured into 3 kg of ice and extracted with Et ₂ O (2 × ₂ L). The combined organic layers are washed with aqueous 1.5N NaOH (2 × 1 L) followed by aqueous saturated NaHCO ₃ (400 mL), or until pH is about 8-9. The organic layer is dried over MgSO ₄ , filtered and concentrated under reduced pressure until constant weight (unreacted 2,6-difluoropyridine is removed: 10-12%). The title compound (207.3 g, 74%) is obtained as a yellow liquid.
[140] (b) 2- (ethylamino) -6-fluoro-3-nitropyridine
[141] To a 2,6-difluoro-3-nitropyridine (45.7 g, 285 mmol) in THF (500 mL) at −40 ° C. is added dropwise a solution of ethylamine (25.7 g, 570 mmol) in THF (250 mL). After 30 minutes, the reaction mixture is concentrated under reduced pressure and the residue is dissolved in EtOAc. The organic phase is washed with brine, dried (MgSO ₄ ), filtered and concentrated. The yellow solid obtained is purified by flash chromatography (15% EtOAc in hexanes) to give the title compound (43.2 g, yield 82%) as a yellow solid.
[142] (c) 3-amino-2- (ethylamino) -6-fluoropyridine
[143] A solution of 2- (ethylamino) -6-fluoro-3-nitropyridine (43.2 g, 230 mmol) in THF (1 L) was added under hydrogen (1 atm) in the presence of 20% Pd (OH) _{2 /} C (4.35 g). ) Stir overnight at room temperature. The catalyst is removed by filtration through diatomaceous earth. The filtrate is concentrated under reduced pressure to afford the title compound (36.3 g, 95% yield) as a black solid.
[144] (d) 2-chloro-N- {2- (ethylamino) -6-fluoro-3-pyridinyl} -5-bromo-3-pyridinecarboxamide
[145] To a cold solution (4 ° C.) of 3-amino-2- (ethylamino) -6-fluoropyridine (31.0 g, 200 mmol) in MeCN (160 mL) was added solid NaHCO ₃ (50.4 g, 600 mmol). After 15 minutes, a solution of 5-bromo-2-chloro-3-pyridinecarbonyl chloride (1 equiv, 200 mmol) in MeCN (155 mL) is added. After 60 minutes at room temperature the reaction mixture is poured into water (1.2 L) and stirred for 30 minutes. The solid obtained is filtered and dried overnight at 50 ° C. under reduced pressure. The title compound (73.7 g, 99% yield) is obtained as a black solid.
[146] (e) 8-bromo-5,11-dihydro-11-ethyl-2-fluoro-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine -6-on
[147] 2-Chloro-N- {2- (ethylamino) -6-fluoro-3-pyridinyl} -5-bromo-3-pyridinecarboxamide (73.5 g, 216 mmol) in pyridine (435 mL) at 50 ° C. ) Is added dropwise to a solution of NaHMDS 1M in THF (520 mL, 520 mmol). After 10 minutes, the reaction is cooled to room temperature and then poured into ice water (2 L). The solid obtained is filtered and rinsed with water followed by hexane. The solid is dried under reduced pressure to afford the title compound (50.6 g, 69% yield) as a dark green solid.
[148] (f) 8-bromo-5,11-dihydro-11-ethyl-2-fluoro-5-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1, 4] diazepine-6-one
[149] 8-Bromo-5,11-dihydro-11-ethyl-2-fluoro-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4 in DMF (520 mL) ] Add NaH (4.28 g, 178 mmol) to a solution of diazepin-6-one (44 g, 130.5 mmol) and heat the mixture to 50 ° C. for 30 minutes. The reaction mixture is cooled to room temperature and treated with MeI (24.4 mL, 522 mmol). After 1.5 hours, the reaction mixture is poured into ice water. The solid is filtered, washed with water then hexanes and dried under reduced pressure to afford the title compound (43.2 g, yield 94%) as a dark gray solid.
[150] (g) 5,11-dihydro-11-ethyl-2-fluoro-5-methyl-8- (2-propenyl) -6H-dipyrido [3,2-b: 2 ', 3'-e ] [1,4] diazepine-6-one
[151] Allyltributyltin (32.0 mL, 103.4 mmol) and Pd (Ph ₃ P) ₄ (5.43 g, 4.70 mmol) were added to 8-bromo-5,11-dihydro-11-ethyl-2 in DMF (470 mL). Degassing solution (solution) of -fluoro-5-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one (33.0 g, 94.0 mmol) Through N ₂ for 45 minutes). An additional amount of Pd (Ph ₃ ) ₄ (1.09 g, 0.94 mmol added after 1, 2, 3, 4, 5 hours of reaction) is added to complete the reaction. The mixture is heated to 90 ° C. for 6 hours. The mixture is concentrated under reduced pressure. The residue is purified by flash chromatography (hexane: EtOAc, 8: 2 to 7: 3) to afford the title compound (22.4 g, 76% yield).
[152] (h) 5,11-dihydro-11-ethyl-2-fluoro-8- (2-hydroxyethyl) -5-methyl-6H-dipyrido [3,2-b: 2 ', 3'- e] [1,4] diazepine-6-one
[153] The ozonated oxygen stream was extracted with 5,11-dihydro-11-ethyl-2-fluoro-5-methyl-8- (2-propenyl) -6H in CH ₂ Cl ₂ (100 mL) and MeOH (100 mL). -Bubble for 3 hours through a cold (-78 ° C) solution of dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one (22.38 g, 71.6 mmol) Ring. The N ₂ stream is then bubbled through the solution for 15 minutes, then solid NaBH ₄ (5.05 g, 133 mmol) is added to the solution. The reaction mixture is allowed to warm up to room temperature. After 1 h, an additional portion of NaBH ₄ (1.62 g, 43.0 mmol) is added to the reaction mixture. After additional time, aqueous saturated NH ₄ Cl (150 mL) is added and the mixture is stirred at room temperature for 30 minutes. The organic solvent is removed under reduced pressure. Water (200 mL) is added and the mixture is extracted with CHCl ₃ (3 × 300 mL). The combined organic layers are dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (EtOAc: CHCl ₃ , 4: 1) to afford the title compound (19.7 g, 72% yield) as a white solid.
[154] (i) 5,11-dihydro-11-ethyl-2-fluoro-5-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[155] Diethyl azodicarboxylate (DEAD) (14.3 mL, 91.0 mmol) was added to 5,11-dihydro-11-ethyl-2-fluoro-8- (2-hydroxyethyl) -5 in THF (300 mL). -Methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one (19.2 g, 60.7 mmol), 4-hydroxyquinoline (13.2 g, 91.0 mmol) and Ph ₃ P (23.9 g, 91.0 mmol) drop wise. The mixture is stirred at rt for 1 h and then concentrated under reduced pressure. The residue was purified by flash chromatography (EtOAc: MeOH, 95: 5) to afford the title compound (17.9 g, yield 67%) as a white solid: MS (ESI) m / z 444 (MH) ⁺ .
[156] Example V (No. 1, Table 1)
[157] 2-chloro-5,11-dihydro-11-ethyl-4-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3 '-e] [1,4] diazepine-6-one
[158] (a) 2-chloro-N- (2,6-dichloro-4-methyl-3-pyridinyl) -3-pyridinecarboxamide
[159] NaHCO ₃ (21.72 g, 258.6 mmol) was added to 3-amino-2,6-dichloro-4-methylpyridine (41.61 g, 235.1 mmol in MeCN (350 mL); KG Grozinger et al. J. Heterocyclic Chem Prepared as described in 1995, 32, 259-263) and the resulting suspension is stirred for 15 minutes. Then a solution of 2-chloronicotinyl chloride (43.44 g, 246.8 mmol) in MeCN (500 mL) is introduced over 30 minutes. The resulting suspension is stirred at room temperature. After 24 hours, the solution appears acidic, so an additional amount of NaHCO ₃ (3.00 g, 35.7 mmol) is added. The suspension is then stirred for another two days at room temperature. The mixture is then poured into a mixture of water (2 L) and ice (200 g) and stirred for 20 minutes. The solid is filtered off, washed with water (500 mL), and the obtained solid is dried under reduced pressure with P ₂ O ₅ to give the title compound (62.55 g, yield 84%) as a white powder.
[160] (b) N- (2,6-dichloro-4-methyl-3-pyridinyl) -2- (ethylamino) -3-pyridinecarboxamide
[161] 2-Chloro-N- (2,6-dichloro-4-methyl-3-pyridinyl) -3-pyridinecarboxamide (63.17 g, 194.0 mmol) and ethylamine (28.0 g, 583 mmol) in xylene (250 mL) The solution is stirred for 7 h at 120-125 ° C. in a strong autoclave. The resulting suspension is poured into water (1 L), stirred for 15 minutes and filtered. The residue is dissolved in EtOAc, washed with water (3 ×), brine and dried over MgSO ₄ . The filtrate is extracted with EtOAc and the combined organic extracts are washed with water and brine and dried over MgSO ₄ . The two fractions are then combined and excess xylene is removed by co-distillation with benzene to afford the title compound (61.98 g, pink solid) as the main component of the mixture of compounds. The material which was not purified at this point is used directly in the subsequent reaction.
[162] (c) 5-bromo-N- (2,6-dichloro-4-methyl-3-pyridinyl) -2- (ethylamino) -3-pyridinecarboxamide
[163] Solid KOAc (22.2 g, 229 mmol) was diluted with crude N- (2,6-dichloro-4-methyl-3-pyridinyl) -2- (ethylamino) -3-pyridinecarboxamide (61.98) in AcOH (600 mL). g, <190 mmol) in solution. Br ₂ (9.7 mL, 191 mmol) solution is added over 30 minutes. After stirring for 30 minutes, additional amounts of KOAc and Br ₂ are introduced until TLC shows no starting material remaining (11.2 g total KOAc and 5.91 mL of Br ₂ ). The solution is then poured into water (2 L) and stirred for 20 minutes. The solid is filtered off and dissolved in Et ₂ O (1 L). The ether solution is washed with water (3 ×), aqueous saturated NaHCO ₃ , brine and then dried (MgSO ₄ ) to give a mixture (77.7 g, yellow foam) containing the title compound as main component. The material is used directly in the subsequent reaction.
[164] (d) 8-bromo-2-chloro-5,11-dihydro-11-ethyl-4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4 Diazepine-6-one
[165] Crude 5-bromo-N- (2,6-dichloro-4-methyl-3-pyridinyl) -2- (ethylamino) -3-pyridinecarboxamide (77.7 g, mmol) in anhydrous pyridine (1.0 L) ) The solution is heated to 50 ° C. The NaHMDS 1M solution in THF (418 mL, 418 mmol) is then added dropwise and stirring is continued for an additional 15 minutes. After cooling to room temperature, water (50 mL) is added and the mixture is concentrated to 2/3 volume on a rotary evaporator. Then, the residue was poured into water (3 L). Filtration affords the title compound (23.44 g) as a tan solid. The filtrate is extracted with EtOAc (3 ×) and the combined extracts are washed with water and brine, then dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (hexane: EtOAc, 9: 1 or CHCl ₃ ) to afford an additional amount of the title compound (14.06 g, 37.50 g in total, 50% yield for 3 steps).
[166] (e) 2-chloro-5,11-dihydro-11-ethyl-4-methyl-8- (2-propenyl) -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[167] 8-Bromo-2-chloro-5,11-dihydro-11-ethyl-4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [in DMF (250 mL) [ 1,4] diazepine-6-one (12.6 g, 34.3 mmol) was degassed under reduced pressure for 20 minutes. Then Pd (PPh ₃ ) ₄ (775 mg, 0.7 mmol) is added followed by allyltributyltin (12.5 mL, 41.1 mmol). After degassing for 10 minutes under reduced pressure, the mixture is heated at 100 ° C. for 7 hours. The mixture is then concentrated under reduced pressure. The residue is purified by flash chromatography (hexane: EtOAc, 19: 1) to give the title compound (8.04 g, 71% yield) as a yellow solid.
[168] (f) 2-chloro-5,11-dihydro-11-ethyl-8- (2-hydroxyethyl) -4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e ] [1,4] diazepine-6-one
[169] ₂ -chloro-5,11-dihydro-11-ethyl-4-methyl-8- (2-propenyl) -6H-dipyrido [3, in CH ₂ Cl ₂ (120 mL) and MeOH (120 mL). Introduce the O ₃ stream into a cold (−78 ° C.) solution of 2-b: 2 ′, 3′-e] [1,4] diazepine-6-one (8.04 g, 24.4 mmol) and Sudan III. If the pink solution turns brown, O ₂ is bubbled through the solution for 10 minutes. NaBH ₄ (1.12 g, 29.4 mmol) is then added and the solution is allowed to warm to room temperature. After 30 minutes, an additional amount of NaBH ₄ (500 mg) is added. After 1 h, aqueous saturated NH ₄ Cl is then added and the mixture is stirred for 20 minutes. The solution is concentrated under reduced pressure and extracted with CH ₂ Cl ₂ (3 ×). The combined organic extracts are washed with brine, dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (CHCl ₃ : EtOH, 97: 3) to afford the title compound (6.01 g, 74% yield) as a white solid.
[170] (g) 2-chloro-5,11-dihydro-11-ethyl-4-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[171] Diethyl azodicarboxylate (DEAD) (83 μl, 0.53 mmol) was added to 2-chloro-5,11-dihydro-11-ethyl-8- (2-hydroxyethyl)-in THF (2.5 mL) at room temperature. 4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (125 mg, 0.38 mmol), 4-hydroxyquinoline (76.3 mg, 0.53 mmol) and Ph ₃ P (138 mg, 0.53 mmol) drop wise. The mixture is stirred at rt for 3 h and then concentrated under reduced pressure. The residue is dissolved in EtOAc (60 mL) and the solution is washed successively with 1N aqueous NaOH (3 × 10 mL) and brine (15 mL), then dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (EtOAc: MeOH, 9: 1) to give the title compound (52 mg, yield 30%) as a white solid: MS (ESI) m / z 460/462 (MH) ⁺ .
[172] Example VI (No. 7, Table 1)
[173] 5,11-dihydro-11-ethyl-2-fluoro-4-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[174] (a) 8-bromo-5,11-dihydro-11-ethyl-2-{{(4-methoxyphenyl) methyl} amino} -4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[175] 8-bromo-2-chloro-5,11-dipyrido-11-ethyl-4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine -6-one (2.50 g, 6.80 mmol) is dissolved in dioxane (10 mL) in a glass pressure bottle and 4-methoxybenzylamine (4.0 mL, 30.6 mmol) is added to the solution. The mixture is heated at 170 ° C. for 5 days. The reaction mixture is cooled to room temperature and concentrated under reduced pressure. The residue is dissolved in EtOAc and washed with 10% aqueous NH ₄ Cl, dried over MgSO ₄ , filtered and concentrated under reduced pressure to give a brown oil which is chromatographed on silica gel (hexanes: EtOAC, 4: 1). . The title compound (1.35 g, yield 42%) is obtained as a yellow solid.
[176] (b) 2-amino-8-bromo-5,11-dihydro-11-ethyl-4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4 Diazepine-6-one
[177] 8-bromo-5,11-dihydro-11-ethyl-2-{{(4-methoxyphenyl) methyl} amino} -4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (6.52 g, 18.7 mmol) is dissolved in trifluoroacetic acid (100 mL) and the solution is magnetically stirred at room temperature for 18 hours. Trifluoroacetic acid is evaporated under reduced pressure and the residue is taken up in a mixture containing water (450 mL), concentrated NH ₄ OH (50 mL) and EtOAc (400 mL). The mixture is magnetically stirred at room temperature for 2 hours. The organic phase is concentrated under reduced pressure. The yellow precipitate is filtered off, washed with water and dried under reduced pressure to afford 7.51 g of a yellow solid which is triturated in Et ₂ O to give the title compound (5.12 g, 105% yield) as a yellow solid.
[178] (c) 8-bromo-5,11-dihydro-11-ethyl-2-fluoro-4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1, 4] diazepine-6-one
[179] 2-amino-8-bromo-5,11-dihydro-11-ethyl-4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4 in plastic bottles ] Diazepine-6-one (5.10 g, 14.6 mmol). HF-pyridine (100 g) is added and the suspension obtained is cooled to 0 ° C. Sodium nitrate (1.12 g, 16.1 mmol) was added in several portions over 30 minutes to form a purple solution. The mixture is then stirred at rt for 16 h. The reaction mixture is poured into ice and 6N NaOH. The beige suspension is extracted with EtOAc. The organic extracts are combined, dried over MgSO ₄ , filtered and concentrated. The residue is triturated with Et ₂ O / hexanes to give the title compound (4.30 g, yield 84%).
[180] (d) 5,11-dihydro-11-ethyl-2-fluoro-4-methyl-8- (2-propenyl) -6H-dipyrido [3,2-b: 2 ', 3'-e ] [1,4] diazepine-6-one
[181] Allyltributyltin (7.77 mL, 25.1 mmol) and Pd (Ph ₃ P) ₄ (1.32 g, 1.14 mmol) were added to 8-bromo-5,11-dihydro-11-ethyl-2 in DMF (114 mL). Degassing solution of -fluoro-4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one (8.0 g, 22.8 mmol) (30 Solution passed through N ₂ for minutes). The mixture is heated to 90 ° C. for 3 hours. The mixture is poured into water (250 mL) and extracted with EtOAc (4 x 250 mL). The combined organic layers are washed with 20% NH ₄ OH (250 mL) and brine (250 mL), dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (hexane: EtOAc, 4: 1 to 1: 1) to afford the title compound (3.16 g, 51% yield).
[182] (e) 5,11-dihydro-11-ethyl-2-fluoro-8- (2-hydroxyethyl) -4-methyl-6H-dipyrido [3,2-b: 2 ', 3'- e] [1,4] diazepine-6-one
[183] The ozonated oxygen stream was subjected to 5,11-dihydro-11-ethyl-2-fluoro-4-methyl-8- (2-propenyl) -6H in CH ₂ Cl ₂ (25 mL) and MeOH (25 mL). -Dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (3.16 g, 10.1 mmol) was bubbled through a cold (-78 ° C) solution. The N ₂ stream is then bubbled through the solution for 15 minutes, then solid NaBH ₄ (669 mg, 17.1 mmol) is added to the solution. The reaction mixture is allowed to warm up to room temperature. After 2 hours, aqueous saturated HN ₄ Cl (50 mL) is added and the mixture is stirred at room temperature for 15 minutes. The organic solvent is removed under reduced pressure. The remaining aqueous solution is extracted with CHCl ₃ (3 × 100 mL). The combined organic layers are dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (EtOAc: CHCl ₃ , 1: 1) to afford the title compound (2.40 g, 75%) as a white solid.
[184] (f) 5,11-dihydro-11-ethyl-2-fluoro-4-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[185] Diisopropyl azodicarboxylate (DIAD) (117 μl, 0.59 mmol) was added to 5,11-dihydro-11-ethyl-2-fluoro-8- (2-hydroxyethyl in THF (1.9 mL) at room temperature. ) -4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (125 mg, 0.40 mmol), 4-hydroxyquinoline (86.0 mg, 0.59 mmol) and Ph ₃ P (156 mg, 0.59 mmol) drop wise. The mixture is stirred at room temperature for 3 hours and then concentrated under reduced pressure. The residue was dissolved in EtOAc (40 mL) and the solution washed successively with 3N aqueous NaOH (3 × 20 mL), aqueous saturated NH ₄ Cl (15 mL), brine (15 mL), and then dried (MgSO ₄ ). Filtration and concentration under reduced pressure. The residue is partially purified by flash chromatography (EtOAc: MeOH, 95: 5). The solid was further purified by reverse phase HPLC (CombiPrep ADS-AQ 50 × 20 mm, 5μ, 120 A, 5-100% MeCN + 0.10% TFA / water + 0.10% TFA, 25 minutes) to give the trifluoro of the title compound as a white solid. Roacetic acid salt (106 mg, yield 48%) is obtained: MS (ESI) m / z 444 (MH) ⁺ .
[186] Example VII (No. 5, Table 1)
[187] 11- (cyclopropyl) -5,11-dihydro-2-fluoro-4-methyl-8- {2- (4-quinolinyloxy) -ethyl} -6H-dipyrido [3,2-b : 2 ', 3'-e] [1,4] diazepine-6-one
[188] (a) 11-cyclopropyl-5,11-dihydro-2-fluoro-8- (2-hydroxyethyl) -4-methyl-6H-dipyrido [3,2-b: 2 ', 3' -e] [1,4] diazepine-6-one
[189] 8-Bromo-2-chloro-11-cyclopropyl-5,11-dihydro-4-methyl-6H-dipyrido [3] using a process analogous to that of the corresponding 11-ethyl homolog described in Example VI. The title compound is prepared from, 2-b: 2 ', 3'-e] [1,4] diazepin-6-one.
[190] (b) 11-cyclopropyl-5,11-dihydro-2-fluoro-4-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b : 2 ', 3'-e] [1,4] diazepine-6-one
[191] Diisopropyl azodicarboxylate (DIAD) (131 μl, 0.66 mmol) was added to 11- (cyclopropyl) -5,11-dihydro-2-fluoro-8- (2- in THF (2.2 mL) at room temperature. Hydroxyethyl) -4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one (140 mg, 0.44 mmol), 4-hydroxy Quinoline (96.4 mg, 0.66 mmol) and Ph ₃ P (124 mg, 0.66 mmol) are added dropwise. The mixture is stirred at rt for 3 h and then concentrated under reduced pressure. The residue was dissolved in EtOAc (60 mL) and the solution was washed successively with 3N aqueous NaOH (3 × 20 mL), aqueous saturated NH ₄ Cl (20 mL) and brine (20 mL), then dried (MgSO ₄ ). Filtration and concentration under reduced pressure. The residue is partially purified by flash chromatography (EtOAc: MeOH, 95: 5). The solid was further purified by reverse phase HPLC (CombiPrep ADS-AQ 50 × 20 mm, 5μ, 120A, 5-100% MeCN + 0.10% TFA / water + 0.10% TFA, 25min) to give the trifluoroacetic acid of the title compound as a white solid. Salt (104 mg, yield 43%) is obtained: MS (ESI) m / z 456 (MH) ⁺ .
[192] Example VIII (No. 8, Table 1)
[193] 5,11-dihydro-11-ethyl-4-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[194] (a) 5,11-dihydro-11-ethyl-8- (2-hydroxyethyl) -4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1, 4] diazepine-6-one
[195] 2-chloro-5,11-dihydro-11-ethyl-8- (2-hydroxyethyl) -4-methyl-6H-dipyrido [3,2-b: 2 ', 3 in EtOH (6 mL) '-e' [1,4] diazepin-6-one (168 mg, 0.51 mmol), ammonium formate (318 mg, 5.1 mmol) and 10% Pd / C (50 mg) mixtures were altered at room temperature. After 24 hours, an additional amount of ammonium formate (200 mg, mmol) and 10% Pd / C (50 mg) is added. After stirring for a further 24 hours, the reaction is concentrated under reduced pressure. The residue is dissolved in CHCl ₃ and the mixture is washed with aqueous saturated NaHCO ₃ , dried (MgSO ₄ ), filtered and concentrated under reduced pressure to give the title compound (122 mg, 81%) as a yellow oil.
[196] (b) 5,11-dihydro-11-ethyl-4-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3' -e] [1,4] diazepine-6-one
[197] Diisopropyl azodicarboxylate (DIAD) (42 μl, 0.21 mmol) was added to 5,11-dihydro-11-ethyl-8- (2-hydroxyethyl) -4-methyl in THF (0.7 mL) at room temperature. -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (42.0 mg, 0.14 mmol), 4-hydroxyquinoline (31 mg, 0.21 mmol) And Ph ₃ P (55.4 mg, 0.21 mmol) dropwise. The mixture is stirred at rt for 2 h and then concentrated under reduced pressure. The residue was dissolved in EtOAc (60 mL) and the solution was washed successively with 3N aqueous NaOH (3 × 20 mL), aqueous saturated NH ₄ Cl (20 mL) and brine (20 mL), dried (MgSO ₄ ), Filter and concentrate under reduced pressure. The residue is partially purified by flash chromatography (EtOAc: MeOH, 95: 5). The solid was further purified by reverse phase HPLC (CombiPrep ADS-AQ 50 × 20 mm, 5μ, 120A, 5-100% MeCN + 0.10% TFA / water + 0.10% TFA, 25min) to give the trifluoro of the title compound as a white solid. Roacetic acid salt (14.2 mg, yield 19%) is obtained: MS (ESI) m / z 426 (MH) ⁺ .
[198] Example IX (No. 3, Table 1)
[199] 11- (cyclopropyl) -5,11-dihydro-4-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3' -e] [1,4] diazepine-6-one
[200] (a) 5-Bromo-2-chloro-N- (2,6-dichloro-4-methyl-3-pyridinyl) -3-pyridinecarboxamide
[201] 3-amino-2,6-dichloro-4-methylpyridine (0.51 g, 2.85 mol) is dissolved in toluene (35 mL) and pyridine (0.27 mL, 3.28 mmol). Then 5-bromo-2-chloro-3-pyridinecarbonyl chloride (0.80 g, 3.14 mmol) is added dropwise over 30 minutes. The resulting mixture is stirred at rt for 1 h, diluted with water and extracted with toluene (2 ×). The combined organic extracts are dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The viscous oil obtained is triturated with CH ₂ Cl ₂ and the white solid is recovered by suction filtration to give the title compound (0.41 g, yield 36%).
[202] (b) 5-bromo-2- (cyclopropylamino) -N- (2,6-dichloro-4-methyl-3-pyridinyl) -3-pyridinecarboxamide
[203] 5-Bromo-2-chloro-N- (2,6-dichloro-4-methyl-3-pyridinyl) -3-pyridinecarboxamide (3.00 g, 7.61 mmol) and cyclopropylamine (2.11 mL) in xylene , 30.5 mmol) is heated in a sealed tube at 120 ° C. for 24 hours. The reaction is heated to 0 ° C., diluted with water and extracted with EtOAc (2 ×). The combined organic extracts are dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The combined residue was purified by softening (Et ₂ O: hexane, 3: 1) to give the title compound (2.35 g, yield 75%) as a pale yellow solid.
[204] (c) 8-bromo-2-chloro-11-cyclopropyl-5,11-dihydro-4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1, 4] diazepine-6-one
[205] 5-Bromo-2- (cyclopropylamino) -N- (2,6-dichloro-4-methyl-3-pyridinyl) -3-pyridinecarboxamide (105 mg, 0.254 mmol) in pyridine (3.0 mL) The solution is heated to 50 ° C. while a NaHMDS 1M solution in THF (0.53 mL, 0.53 mmol) is added. The resulting solution is stirred at 50 ° C. for 3 hours. The reaction is quenched with aqueous saturated NH ₄ Cl, diluted with water and extracted with EtOAc (3 ×). The combined organic extracts are washed with brine, dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The obtained residue was purified by flash chromatography (hexane: EtOAc, 7: 3) to give the title compound (32 mg, yield 33%).
[206] (d) 2-chloro-11-cyclopropyl-5,11-dihydro-4-methyl-8- (2-propenyl) -6H-dipyrido [3,2-b: 2 ', 3'-e ] [1,4] diazepine-6-one
[207] 8-Bromo-2-chloro-11-cyclopropyl-5,11-dihydro-4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] in DMF (2.0 mL) A [1,4] diazepine-6-one (0.046 g, 0.122 mmol) solution is degassed by sparging with N ₂ for 10 minutes. Pd (PPh ₃ ) ₄ (2.8 mg, 0.003 mmol) and allyltributyltin (0.45 mL, 0.146 mmol) are added and the resulting solution is heated to 90 ° C. for 2.5 h. The reaction mixture is diluted with water and CH ₂ Cl Extract with ₂ (3 ×) and EtOAc (2 ×). The combined organic extracts are dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The obtained residue was purified by flash chromatography (hexane: EtOAc, 7: 3) to give the title compound (0.033 g, yield 80%).
[208] (e) 2-chloro-11-cyclopropyl-5,11-dihydro-8- (2-hydroxyethyl) -4-methyl-6H-dipyrido [3,2-b: 2 ', 3'- e] [1,4] diazepine-6-one
[209] ₂ -Chloro-11-cyclopropyl-5,11-dihydro-4-methyl-8- (2-propenyl) -6H-dipyrido [3 in CH ₂ Cl ₂ (7.5 mL) and MeOH (7.5 mL) [3]. A cold (-78 ° C) solution of, 2-b: 2 ', 3'-e] [1,4] diazepine-6-one (0.82 g, 2.43 mmol) was diluted with O ₃ until the solution turned pale blue. Saturate. The reaction mixture is left at −78 ° C. for 10 minutes, then NaBH ₄ (0.23 g, 9.71 mmol) is added and the mixture is allowed to slowly warm over 2 hours at room temperature. The reaction is quenched with 10% aqueous citric acid solution and extracted with EtOAc (3 ×). The organic extracts are combined, washed with brine, dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The obtained residue was purified by flash chromatography (MeOH: CH ₂ Cl ₂ , 2 to 10%) to give the title compound (0.41 g, yield 60%) as a white solid.
[210] (f) 11-cyclopropyl-5,11-dihydro-8- (2-hydroxyethyl) -4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1 , 4] diazepine-6-one
[211] The title compound is prepared using a method similar to that described for the 11-ethyl homolog of Example VIIIa.
[212] (g) 11-cyclopropyl-5,11-dihydro-4-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3 '-e] [1,4] diazepine-6-one
[213] Diisopropyl azodicarboxylate (DIAD) (167 μl, 0.85 mmol) was added to 11-cyclopropyl-5,11-dihydro-8- (2-hydroxyethyl) -4- in THF (2.8 mL) at room temperature. Methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (175 mg, 0.57 mmol), 4-hydroxyquinoline (123 mg, 0.85 mmol) And Ph ₃ P (223 mg, 0.85 mmol) dropwise. The mixture is stirred at rt for 3 h and then concentrated under reduced pressure. The residue was dissolved in EtOAc (60 mL) and the solution was washed successively with 3N aqueous NaOH (3 × 20 mL), aqueous saturated NH ₄ Cl (20 mL) and brine (20 mL), then dried (MgSO ₄ ). , Filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (EtOAc: MeOH, 95: 5) to give the title compound (74 mg, yield 30%) as a white solid: MS (ESI) m / z 438 (MH) ⁺ .
[214] Example X (No. 21, Table 1)
[215] 5,11-dihydro-11-ethyl-5-methyl-8- {2-{(1-oxido-4-quinolinyl) oxy} ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[216] Solid mCPBA (80-85%, m-chloroperbenzoic acid) (2.21 g, 10.2 mmol) was added 5,11-dihydro-11-ethyl- in CH ₂ Cl ₂ (30 mL) and THF (30 mL) at room temperature. 5-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one (2.56 g, 6.02 mmol) is added to the solution (note that the reaction can also be carried out with CH ₂ Cl ₂ alone). The mixture is stirred at rt for 1 h and then concentrated under reduced pressure. The residue was dissolved in CHCl ₃ (400 mL), the solution was washed successively with 10% Na ₂ S ₂ 0 ₃ (3 × 75 mL), aqueous saturated NaHCO ₃ (3 × 75 mL) and brine (75 mL). , Dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (CHCl ₃ : MeOH, 9: 1) to give the title compound (2.01 g, yield 76%) as a white solid: MS (ESI) m / z 442 (MH) ⁺ .
[217] Example XI (No. 17, Table 1)
[218] 2-chloro-5,11-dihydro-11-ethyl-5-methyl-8- {2-{(1-oxido-4-quinolinyl) oxy} ethyl} -6H-dipyrido [3,2 -b: 2 ', 3'-e] [1,4] diazepine-6-one
[219] Using the same process as in Example X, 2-chloro-5,11-dihydro-11-ethyl-5-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [ From 3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one (2.70 g, 5.87 mmol) the title compound (2.05 g, yield 73%) is obtained as a white solid: MS (ESI) m / z 476/478 (M−H) ⁺ .
[220] Example XII (No. 11, Table 1)
[221] 5,11-dihydro-11-ethyl-2-fluoro-5-methyl-8- {2-{(1-oxido-4-quinolinyl) oxy} ethyl} -6H-dipyrido [3, 2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[222] Solid mCPBA (457 mg, 2.12 mmol) was added to 5,11-dihydro 2-fluoro-11-ethyl-5-methyl-8- {2- (4-quinolinyloxy) in CH ₂ Cl ₂ (10 mL). Ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (470 mg, 1.06 mmol) was added to the solution. The mixture is stirred at rt for 13 h and then concentrated under reduced pressure. The residue was dissolved in CHCl ₃ (75 mL) and the solution was successively washed with aqueous 10% Na ₂ S ₂ 0 ₃ (3 × 25 mL), aqueous saturated NaHCO ₃ (3 × 25 mL) and brine (25 mL). Wash, dry (MgSO ₄ ), filter and concentrate under reduced pressure. The residue was flash chromatography (CHCl ₃ : MeOH, 19: 1 to 9: 1) to give the title compound (250 mg, yield 51%) as a white solid: MS (ESI) m / z 460 (MH) ⁺ .
[223] Example XIII (No. 10, Table 1)
[224] 2-Chloro-11-cyclopropyl-5,11-dihydro-5-methyl-8- {2-{(1-oxydo-4-quinolinyl) oxy} ethyl} -6H-dipyrido [3, 2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[225] Solid mCPBA (177 mg, 0.82 mmol) was added 2-chloro-11-cyclopropyl-5,11-dihydro-5-methyl-8- {2- () in CH ₂ Cl ₂ (3 mL) and THF (3 mL). To 4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (143 mg, 0.30 mmol) solution . The mixture is stirred at rt for 14 h and then concentrated under reduced pressure. The residue was dissolved in EtOAc (50 mL) and the solution washed successively with aqueous 10% Na ₂ S ₂ 0 ₃ (3 × 10 mL), aqueous saturated NaHCO ₃ (3 × 10 mL) and brine (10 mL). , Dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue is partially purified by flash chromatography (EtOAc: MeOH, 95: 5 to CH ₂ Cl ₂ : MeOH, 9: 1). The impure compound is further purified by reverse phase HPLC (CombiPrep ADS-AQ 50 × 20 mm, 5μ, 120 A, 5-100% MeCN + 0.10% TFA / water + 0.10% TFA, 25 min). The pure fractions are treated with aqueous saturated NaHCO ₃ , extracted with EtOAc, the solution is dried (MgSO ₄ ), filtered and concentrated under reduced pressure to give the title compound (28.3 mg, 19%) as a white solid: MS ( ESI) m / z 488/490 (MH) ⁺ .
[226] Example XIV (No. 14, Table 1)
[227] 2-chloro-5,11-dihydro-11-ethyl-4-methyl-8- {2-{(1-oxido-4-quinolinyl) oxy} ethyl} -6H-dipyrido [3,2 -b: 2 ', 3'-e] [1,4] diazepine-6-one
[228] Solid mCPBA (46.9 mg, 0.22 mmol) was added 2-chloro-5,11-dihydro-ll-ethyl-5-methyl-8- {2- (4-quinolinyloxy in CH ₂ Cl ₂ (2.2 mL). ) Ethyl) -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (50.0 mg, 0.11 mmol) solution. The mixture is stirred at rt for 1 h and then concentrated under reduced pressure. The residue was dissolved in EtOAc (50 mL) and the solution washed successively with aqueous 10% Na ₂ S ₂ 0 ₃ (3 × 10 mL), aqueous saturated NaHCO ₃ (3 × 10 mL) and brine (10 mL). , Dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue is purified by reverse phase HPLC (CombiPrep ADS-AQ 50 × 20 mm, 5, 120 A, 5-10% MeCN + 0.10% TFA / water + 0.10% TFA, 25 min). The pure fractions were treated with aqueous saturated NaHCO ₃ , extracted with EtOAc, the solution was dried (MgSO ₄ ), filtered and concentrated under reduced pressure to afford the title compound (32.5 mg, 63%) as a white solid: MS ( ESI) m / z 488/490 (M−H) ⁺ .
[229] Example XV (No. 9, Table 1)
[230] 5,11-dihydro-11-ethyl-2-fluoro-4-methyl-8- {2-{(1-oxido-4-quinolinyl) oxy} ethyl} -6H-dipyrido [3, 2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[231] 5,11-dihydro-11-ethyl-2-fluoro-4-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-, using a process similar to that of Example XIV. From the dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one (125 mg, 0.28 mmol) the title compound (40 mg, 31%) is obtained as a white solid: MS (ESI) m / z 460 (MH) ⁺ .
[232] Example XVI (No. 12, Table 1)
[233] 5,11-Dihydro-11-ethyl-4-methyl-8- {2-{(1-oxido-4-quinolinyl) -oxy} ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[234] Solid mCPBA (163 mg, 0.75 mmol) was added to 5,11-dihydro-11-ethyl-4-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H in CH ₂ Cl ₂ (7.5 mL). -Dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (144 mg, 0.34 mmol) in solution. The solution is stirred at room temperature for 2 hours. The mixture is diluted with CH ₂ Cl ₂ (125 mL) and washed successively with aqueous 10% Na ₂ S ₂ 0 ₃ (2 × 25 mL), aqueous saturated NaHCO ₃ (2 × 25 mL) and brine (25 mL). Then it is dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (CH ₂ Cl ₂ : MeOH, 10: 1) to afford the title compound (102 mg, yield 68%) as a white solid: MS (ESI) m / z 442 (MH).
[235] Example XVII (No. 15, Table 1)
[236] 11-cyclopropyl-5,11-dihydro-11-ethyl-4-methyl-8- {2-{(l-oxido-4-quinolinyl) oxy} ethyl} -6H-dipyrido [3, 2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[237] 11-cyclopropyl-5,11-dihydro-11-4-methyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido, using a process similar to that of Example XIV. From [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (37.1 mg, 0.085 mmol) the title compound (35 mg, 91%) is obtained as a white solid: MS (ESI) m / z 454 (M−H) ⁺ .
[238] Example XVIII (No. 25, Table 1)
[239] 2-chloro-5,11-dihydro-11-ethyl-4-methyl-8- {2- (5-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3 '-e] [1,4] diazepine-6-one
[240] Diethyl azodicarboxylate (DEAD) (77 μl, 0.49 mmol) was added 2-chloro-5,11-dihydro-11-ethyl-8- (2-hydroxyethyl)-in THF (2.5 mL) at room temperature. 4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (125 mg, 0.38 mmol), 5-hydroxyquinoline (70.9 mg, 0.49 mmol) and Ph ₃ P (128 mg, 0.49 mmol) drop wise. After 1.5 hours, an additional amount of DEAD (35 μl, 0.22 mmol) and PPh ₃ (59 mg, 0.22 mmol) is added to the mixture. The mixture was stirred at rt for 1 h, then diluted in EtOAc (50 mL), the solution was washed successively with aqueous 1N NaOH (4 x 10 mL) and brine (15 mL), dried (MgSO ₄ ), Filter and concentrate under reduced pressure. The residue is purified by flash chromatography (CH ₂ Cl ₂ : acetone, 85:15). The solid obtained was triturated with Et ₂ 0 to afford the title compound (38 mg, yield 22%) as a white solid: MS (ESI) m / z 460/462 (MH) ⁺ .
[241] Example XIX (No. 20, Table 1)
[242] 2-chloro-5,11-dihydro-11-ethyl-5-methyl-8- (2-{(1-oxido-5-quinolinyl) oxy} ethyl} -6H-dipyrido [3,2 -b: 2 ', 3'-e] [1,4] diazepine-6-one
[243] Diethyl azodicarboxylate (DEAD) (160 μl, 1.01 mmol) was added 2-chloro-5,11-dihydro-11-ethyl-8- (2-hydroxyethyl)-in THF (3.4 mL) at room temperature. 5-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (225 mg, 0.68 mmol), 5-hydroxyquinoline (147 mg, 1.01 mmol) and Ph ₃ P (266 mg, 1.01 mmol) dropwise. The mixture is stirred at rt for 3 h and then concentrated under reduced pressure. The residue is partially purified by flash chromatography (EtOAc: MeOH, 95: 5). Fractions containing 5-quinolinyloxy derivatives are concentrated, dissolved in CH ₂ Cl ₂ (2 mL) and THF (5 mL) and treated with mCPBA (80%, 248 mg, 1.15 mmol) at room temperature. The solution is stirred at room temperature for 2.5 hours and then concentrated under reduced pressure. The residue was dissolved in EtOAc (50 mL) and washed successively with aqueous 10% Na ₂ S ₂ 0 ₃ (3 × 10 mL), aqueous saturated NaHCO ₃ (3 × 10 mL) and brine (10 mL), Dry (MgSO ₄ ), filter and concentrate under reduced pressure. The residue was purified by flash chromatography (EtOAc: MeOH, 95: 5 to CH ₂ Cl ₂ : MeOH, 9: 1) to afford the title compound (113 mg, 35% yield for two steps) as a white solid: MS ( ESI) m / z 476/478 (M−H) ⁺ .
[244] Example XX (No. 18, Table 1)
[245] 5,11-dihydro-11-ethyl-2-fluoro-5-methyl-8- {2-{(1-oxy-5-quinolinyl) oxy} ethyl} -6H-dipyrido [3, 2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[246] Using the process of Example XIX, 5,11-dihydro-11-ethyl-2-fluoro-8- (2-hydroxyethyl) -5-methyl-6H-dipyrido [3,2-b: From 2 ', 3'-e] [1,4] diazepine-6-one (267 mg, 0.84 mmol), the title compound (204 mg, yield 52% for two steps) is obtained as a white solid: MS (ESI). m / z 460 (M−H) ⁺ .
[247] Example XXI (No. 19, Table 1)
[248] 2-chloro-5,11-dihydro-11-ethyl-4-methyl-8- {2-{(1-oxido-5-quinolinyl) oxy} ethyl} -6H-dipyrido [3,2 -b: 2 ', 3'-el [1,4] diazepine-6-one
[249] 2-Chloro-5,11-dihydro-11-ethyl, using the same process as in Example XIX, except diisopropyl azodicarboxylate (DIAD) was used instead of diethyl azodicarboxylate -8- (2-hydroxyethyl) -4-methyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one (223 mg, 0.67 mmol ) Gives the title compound (43.4 mg, yield 14% for 2 steps) as a white solid: MS (ESI) m / z 476/478 (MH) ⁺ .
[250] Example XXII (No. 23, Table 1)
[251] 5,11-dihydro-11-ethyl-2-fluoro-4-methyl-8- {2-{(1-oxido-5-quinolinyl) oxy} ethyl} -6H-dipyrido [3, 2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[252] 5,11-Dihydro-11-ethyl-2-fluoro-8- (2-hydroxyethyl) -4-methyl-6H-dipyrido [3,2 using a process similar to the process of Example XXI -b: 2 ', 3'-e] [1,4] diazepine-6-one (250 mg, 0.79 mmol) affords the title compound (64.7 mg, yield 18% for two steps) as a white solid: MS (ESI) m / z 460 (MH) ⁺ .
[253] Example XXIII (No. 22, Table 1)
[254] 2-chloro-5,11-dihydro-11-ethyl-8- {2-{(1-oxido-4-quinolinyl) oxy} ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[255] MS (ESI) m / z 462/464 (M−H) ⁺ .
[256] Example XXIV (No. 26, Table 1)
[257] 5,11-dihydro-5,11-diethyl-8- (2- (4-quinolinyloxy) ethyl) -6H-dipyrido [3,2-b: 2 ', 3'-e] [ 1,4] diazepine-6-one
[258] MS (ESI) m / z 440 (MH) ⁺ .
[259] Example XXV (No. 27, Table 1)
[260] 5,11-dihydro-5,11-diethyl-8- {2-{(1-oxido-4-quinolinyl) oxy} ethyl} -6H-dipyrido [3,2-b: 2 ' , 3'-e] [1,4] diazepine-6-one
[261] MS (ESI) m / z 456 (MH) ⁺ .
[262] Example XXVI (No. 28, Table 1)
[263] 2,5-dimethyl-5,11-dihydro-11-ethyl-8- {2-{(1-oxido-4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[264] (a) 5-Bromo-2-chloro-N- (6-chloro-2-methyl-3-pyridinyl) -3-pyridinecarboxamide
[265] NaHCO ₃ (3.9 g, 46.4 mmol) was added 3-amino-2-chloro-6-methylpyridine [2.2 g, 15.4 mmol in MeCN (50 mL); (KG, Grozinger et al., J. Heterocyclic Chem. 1995, 32, 259-263). The resulting suspension was stirred for 15 minutes and crude 5-bromo-2-chloro-3-pyridinecarbonyl chloride in MeCN (10 mL), except omitting the aqueous workup, see TW Gero prepared from 5-bromo-2-hydroxy-3-pyridinecarboxylic acid and SOCl ₂ , as described in et al., Synth.Commun. 1989, 19, 553-559 (incorporated herein by reference). The solution is introduced over 30 minutes. The resulting suspension is stirred at room temperature. After 24 hours, the mixture is poured into a mixture of water (100 mL) and ice (10 g) and stirred for 20 minutes. The suspension is filtered and the solid obtained is washed with water (50 mL) and hexane (25 mL) and then dried under reduced pressure with P ₂ 0 ₅ to afford the title compound (1.8 g, 31% yield) as a white powder.
[266] (b) 5-bromo-N- (2-chloro-6-methyl-3-pyridinyl) -2- (ethylamino) -3-pyridinecarboxamide
[267] 5-Bromo-2-chloro-N- (2-chloro-6-methyl-3-pyridinyl) -3-pyridinecarboxamide (1.7 g, 4.8 mmol) and ethylamine (THF) in THF (5 mL) 2M, 10.5 mmol, 5.2 mL) solution is stirred for 16 h at 90-95 ° C. in a steel autoclave. The resulting mixture was poured into water (100 mL), stirred for 15 minutes and filtered. The solid is washed with water then hexanes to give the title compound (1.58 g, yield 89%) as a yellow solid.
[268] (c) 8-bromo-5,11-dihydro-2-5-dimethyl-11-ethyl-6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] dia Zepin-6-on
[269] Crude 5-bromo-N- (2-dichloro-6-methyl-3-pyridinyl) -2- (ethylamino) -3-pyridinecarboxamide (0.7 g, 1.8 mmol) in anhydrous pyridine (18 mL) Is heated to 50 ° C. NaHMDS 1M solution in THF (7.2 mL, 7.2 mmol) was added dropwise and stirring continued for an additional 3 hours. After cooling to room temperature, methyl iodide (0.6 mL, 9 mmol) is added and the reaction is stirred overnight. Water (25 mL) is added and the mixture is extracted with EtOAc (3 ×) and the combined extracts are washed with water and brine, dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (hexane: EtOAc, 8: 2) to afford the title compound (259 mg, yield 41%).
[270] (d) 5,11-dihydro-2,5-dimethyl-11-ethyl-8- (2-propenyl) -6H-dipyrido [3,2-b: 2 ', 3'-e] [1 , 4] diazepine-6-one
[271] 8-Bromo-5,11-dihydro-2,5-dimethyl-11-ethyl-6H-dipyrido [3,2b: 2 ', 3'-e] [1,4] in DMF (7.4 mL) Diazepine-6-one (258 mg, 0.7 mmol) solution is degassed under reduced pressure for 20 minutes. Then Pd (PPh ₃ ) ₄ (43 mg, 0.04 mmol) is added followed by allyltributyltin (0.3 mL, 0.85 mmol). After degassing under reduced pressure for 10 minutes, the mixture is heated to 100 ° C. for 1.5 hours. . The mixture is then concentrated under reduced pressure. The residue is purified by flash chromatography (hexane: EtOAc, 8: 2) to give the title compound (184 mg, yield 98%) as a yellow solid.
[272] (e) 5,11-dihydro-2,5-dimethyl-11-ethyl-8- (2-hydroxyethyl) -6H-dipyrido [3,2-b: 2 ', 3'-e] [ 1,4] diazepine-6-one
[273] The O ₃ stream was extracted with 5,11-dihydro-2,5-dimethyl-11-ethyl-8- (2-propenyl) -6H-dipyrido in CH ₂ Cl ₂ (3 mL) and MeOH (3 mL). 3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (187 mg, 0.6 mmol) and a cold (-78 ° C.) solution of Sudan III. If the pink solution turns brown, 0 ₂ is bubbled through the solution for 10 minutes. NaBH ₄ (57 mg, 1.52 mmol) is added and the solution is allowed to warm to room temperature. After 30 minutes, an additional amount of NaBH ₄ (20 mg) is added. After 2 hours, aqueous saturated NH ₄ Cl is added and the mixture is stirred for 20 minutes. The solution is concentrated under reduced pressure and extracted with CH ₂ Cl ₂ (3 ×). The combined organic extracts are washed with brine, dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (EtOAc: hexanes 6: 4) to afford the title compound (111 mg, yield 58%) as a white solid.
[274] (f) 2,5-dimethyl-5,11-dihydro-11-ethyl-8- {2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[275] Diethyl azodicarboxylate (DEAD) (139 μl, 0.88 mmol) was added to 5,11-dihydro-2,5-dimethyl-11-ethyl-8- (2-hydroxyethyl in THF (3.5 mL) at room temperature. ) -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (110 mg, 0.35 mmol), 4-hydroxyquinoline (128 mg, 0.88 mmol) And Ph ₃ P (231 mg, 0.88 mmol) dropwise. The mixture is stirred at rt for 16 h. The reaction mixture is diluted in EtOAc (60 mL) and the solution is washed successively with 1N aqueous NaOH (3 × 10 mL) and brine (15 mL), then dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (EtOAc then EtOAc: MeOH, 9.5: 0.5) to give the title compound (57 mg, yield 37%) as a white solid: MS (ESI) m / z 440 (MH) ⁺ .
[276] (g) 5,11-dihydro-2,5-dimethyl-11-ethyl-8- {2-{(1-oxydo-4-quinolinyl) oxy} ethyl} -6H-dipyrido [3, 2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[277] Solid mCPBA (80-85%, m-chloroperbenzoic acid) (60 mg, 0.28 mmol) was added 5,11-dihydro-2,5-dimethyl-11-ethyl-8 in CH ₂ Cl ₂ (1.3 mL) at room temperature. -{2- (4-quinolinyloxy) ethyl} -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one (55 mg, 0.13 mmol ) Is added to the solution. The mixture is stirred at rt for 2.5 h and then diluted with CH ₂ Cl ₂ . The resulting solution is washed successively with aqueous 10% Na ₂ S ₂ O ₃ , aqueous saturated NaHCO ₃ and brine, dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (MeOH / EtOAc, 2% to 5%) to afford the title compound (59 mg, yield 99%) as a white solid: MS (ESI) m / z 456 (MH) ⁺ .
[278] Example XXVII (No. 29, Table 1)
[279] 5,11-Dihydro-11-ethyl-5-methyl-8- {2- (1-oxo-4-quinolinyloxy) ethyl} -2- (trifluoromethyl) -6H-dipyrido [3 , 2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[280] (a) 2- (ethylamino) -3-nitro-6- (trifluoromethyl) pyridine
[281] To water (165 mL) 2-nitroacetamide ammonium salt (4 g, 33 mmol) was added piperidinium acetate (33 mmol) in water and then 4-ethoxy-1,1,1-trifluoro in MeOH (11 mL). 3-buten-2-one (6.1 mL, 43 mmol) was added slowly. The reaction is stirred at room temperature for 2 hours and at reflux for 3 hours. The reaction mixture is cooled to 45 ° C. and aqueous HCl (1N) is added until the pH is acidic. After 1 hour at room temperature, the reaction mixture is extracted with CH ₂ Cl ₂ (3 ×). The combined organic layers are washed successively with water and brine, dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (EtOAc then EtOAC: MeOH, 9: 1) to give 3-nitro-6- (trifluoromethyl) -2 (1H) -pyridinone (2.7 g, 40% yield) as a yellow solid. ).
[282] Sodium hydride (350 mg, 13.8 mmol) is added to 3-nitro-6- (trifluoromethyl) -2 (1H) -pyridinone (2.4 g, 11.5 mmol) in DMF (69 mL). The reaction is stirred at 45 ° C. for 30 minutes, cooled to room temperature and N-phenyltrifluoromethanesulfonimide (4.9 g, 13.8 mmol) is added. After 1 hour, 2M ethylamine solution in THF (13 mL, 25 mmol) is added. The reaction is stirred overnight at room temperature. The reaction mixture is poured into water (100 mL) and extracted with EtOAc (3 ×). The combined organic layers are washed successively with water and brine, dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (hexane: EtOAc, 9: 1) to give the desired compound as a yellow oil (1.1 g, 41% yield).
[283] (b) 3-amino-2- (ethylamino) -6- (trifluoromethyl) pyridine
[284] A solution of 2- (ethylamino) -3-nitro-6- (trifluoromethyl) pyridine (1.1 g, 4.9 mmol) in MeOH (40 mL) in the presence of 20% Pd (OH) ₂ / C (100 mg) Stir overnight at room temperature under hydrogen (1 atm). The catalyst is removed by filtration through diatomaceous earth. The filtrate is concentrated under reduced pressure to give the title compound (1 g) as an orange oil.
[285] (c) 2-chloro-N- {2- (ethylamino) -6- (trifluoromethyl) -3-pyridinyl} -5-bromo-3- pyridinecarboxamide
[286] To a 3-amino-2- (ethylamino) -6-trifluoromethyl pyridine (1 g, 4.9 mmol) cold solution in MeCN (24 mL) was added solid NaHC0 ₃ (906 mg, 11 mmol). After 5 minutes, prepared from crude 5-bromo-2-chloro-3-pyridinecarbonyl chloride [5-bromo-2-hydroxy-3-pyridinecarboxylic acid and SOCl ₂ {except for aqueous workup) As described in TW Gero et al. In Synth.Commun. 1989, 19, 553-559 (incorporated herein by reference)) (1 equivalent, 4.9 mmol). After 2 hours, the reaction mixture is poured into ice / H ₂ O (1.5 L) and the solid obtained is filtered and rinsed with H ₂ O followed by hexane. After drying under reduced pressure overnight, the title compound (1.6 g, yield 77%) is obtained as a light brown solid.
[287] (d) 8-bromo-5,11-dihydro-11-ethyl-2- (trifluoromethyl) -6H-dipyrido [3,2-b: 2 ', 3'-e] [1, 4] diazepine-6-one
[288] 2-chloro-N- {2- (ethylamino) -6- (trifluoromethyl) -3-pyridinyl} -5-bromo-3-pyridinecarboxamide (766 mg, 1.8 in DMF (18 mL)) mmol) was added to the solution. NaH (137 mg, 5.4 mmol) was added. The reaction is stirred at 80 ° C. After 1 hour, the reaction is cooled to room temperature and then poured into water (20 mL). The resulting mixture is extracted with EtOAc (3 ×). The combined organic layers are washed successively with water and brine, then dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (hexane: EtOAc, 7: 3) to afford the desired compound as a yellow solid (200 mg, 28% yield).
[289] (e) 8-bromo-5,11-dihydro-11-ethyl-5-methyl-2- (trifluoromethyl) -6H-dipyrido [3,2-b: 2 ', 3'-e ] [1,4] diazepine-6-one
[290] 8-Bromo-5,11-dihydro-11-ethyl-2- (trifluoromethyl) -6H-dipyrido [3,2-b: 2 ', 3'-e] in DMF (2.6 mL) To a [1,4] diazepine-6-one (200 mg, 0.52 mmol) solution is added NaH (20 mg, 0.78 mmol) and the mixture is heated at 50 ° C. for 30 minutes. The reaction mixture is cooled to room temperature and treated with MeI (97 μl, 1.56 mmol). After 16 hours, the reaction mixture is diluted with water. The resulting mixture is extracted with EtOAc (3 ×). The combined organic layers are washed successively with water and brine, dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (hexane: EtOAc, 8.5: 1.5) to give the title compound (110 mg, yield 53%) as a white foam.
[291] (f) 5,11-dihydro-11-ethyl-5-methyl-8- (2-propenyl) -2- (trifluoromethyl) -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[292] Allyltributyltin (98 μl, 0.32 mmol) and Pd (Ph ₃ P) ₄ (16 mg, 0.02 mmol) were added to 8-bromo-5,11-dihydro-11-ethyl- in DMF (1.4 mL) at room temperature. Of 5-methyl-2- (trifluoromethyl) -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one (11 mg, 0.27 mmol) To degassed solution (passed N ₂ for 30 min through solution). The mixture is stirred at 90 ° C. for 1.5 h, then cooled to rt and concentrated under reduced pressure. The residue is purified by flash chromatography (hexane: EtOAc, 8: 2 to 7: 3) to afford the title compound (101 mg, 99% yield).
[293] (g) 5,11-dihydro-11-ethyl-8- (2-hydroxyethyl) -5-methyl-2- (trifluoromethyl) -6H-dipyrido [3,2-b: 2 ' , 3'-e] [1,4] diazepine-6-one
[294] The ozonated oxygen stream was extracted with 5,11-dihydro-11-ethyl-5-methyl-8- (2-propenyl) -2- (trifluoro) in CH ₂ Cl ₂ (2.7 mL) and MeOH (2.7 mL). Methyl) -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one (100 mg, 0.27 mmol) through a cold (-78 ° C.) solution of 2.5 Bubble for time. The N ₂ stream is then bubbled through the solution for 15 minutes, then solid NaBH ₄ (52 mg, 1.4 mmol) is added to the solution. The reaction mixture is allowed to warm up to room temperature. After 1 h, aqueous saturated NH ₄ Cl (5 mL) is added and the mixture is stirred at rt for 2 h. The reaction mixture is diluted with CH ₂ Cl ₂ and the aqueous layer is reextracted with CH ₂ Cl ₂ . The combined organic layers are washed successively with water and brine, dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue is purified by flash chromatography (EtOAc: hexane, 75:25 to EtOAc 100%) to give the title compound (60 mg, yield 59%) as a white solid.
[295] (h) 5,11-dihydro-11-ethyl-5-methyl-8- {2- (4-quinolinyloxy) ethyl} -2- (trifluoromethyl) -6H-dipyrido [3, 2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[296] Diethyl azodicarboxylate (DEAD) (38 μl, 0.24 mmol) was added to 5,11-dihydro-11-ethyl-8- (2-hydroxyethyl) -5-methyl- in THF (1.6 mL) at room temperature. 2- (trifluoromethyl) -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepin-6-one (58 mg, 0.16 mmol), 4-hydroxy Add dropwise to quinoline (35 mg, 0.24 mmol) and Ph ₃ P (62 g, 0.24 mmol) solution. The mixture is stirred at rt for 1 h and then concentrated under reduced pressure. The residue was purified by flash chromatography (EtOAc: MeOH; 95: 5) to give the title compound (24 mg, 31% yield) as a white solid: MS (ESI) m / z 494, 495 (MH) ⁺ .
[297] (i) 5,11-dihydro-11-ethyl-5-methyl-8- {2-{(1-oxydo-4-quinolinyl) oxy} ethyl} -2- (trifluoromethyl)- 6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6-one
[298] Solid mCPBA (80-85%, m-chloroperbenzoic acid) (16 mg, 0.08 mmol) was added 5,11-dihydro-11-ethyl-5-methyl-8- {in CH ₂ Cl ₂ (0.5 mL) at room temperature. 2- (4-quinolinyloxy) ethyl} -2- (trifluoromethyl) -6H-dipyrido [3,2-b: 2 ', 3'-e] [1,4] diazepine-6 Add to warm (22 mg, 0.04 mmol) solution. The mixture is stirred at rt for 2.5 h and then diluted with CH ₂ Cl ₂ . The resulting solution is washed successively with aqueous 10% Na ₂ S ₂ O ₃ , aqueous saturated NaHCO ₃ and brine, dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (MeOH: CHCl ₃ , 5% to 10%) to afford the title compound (11 mg, yield 52%) as a white solid: MS (ESI) m / z 511 (MH) ⁺ .
[299]
[300]
[301]
[302]
[303] Reverse transcription (RT) test
[304] Test theory:
[305] Among the enzymes encoded by human immunodeficiency virus (HIV-1), reverse transcriptases are named because they transcribe DNA copies from RNA templates. This activity can be quantitatively measured in acellular enzyme assays and synthetic template polys with biotinylated oligo d (G) as primers to transcribe radiolabeled DNA strands where reverse transcriptase uses 3H-dGTP as a substrate. Based on the observation that r (C) can be used. The assays described below use wild type enzymes (which are the predominant enzyme forms observed in patients infected with HIV-1), and also under similar assay conditions tyrosine residues at mutant RT enzymes (eg, Y181C, codon 181 are cysteine residues). Prepared by site-directed mutagenesis). This assay allows the compound to be evaluated for its effectiveness in inhibiting mutant cells.
[306] matter:
[307] (a) Preparation of enzyme
[308] Some HIV-1 IIIB clone BH10 RT mutants were seeded in vector pKK233-2 (Pharmacia). -K. Provided by Dr. C.-K. Shih; Boehringer Ingelheim Pharmaceuticals, Inc., U.S.A. The HIV RT clone pKRT2, which contains only the RT p66 gene, regulated briefly by the lac operon / trc promoter, is W. Dr. Summers [Dr. W. Summers (Yale University). Various specific amino acid substituents are introduced into the wild type RT gene by site directed mutagenesis. RT clones are subcloned into pKK233-2 bacterial expression vector. Provided clones include wild type Val106Ala, Tyr181Cys, Tyr188Cys, Tyr188Leu, Gly190Ala and Pro236Leu. Others are made in vivo by site directed mutagenesis of pKK233-2 RT clones including Lys103Asn, Lys103Asn / Tyr181Cys, Lys103Asn / Leu100Ile, Lys103Asn / Pro225His and Lys103Asn / Val108Ile.
[309] (b) Purification of Enzymes
[310] Purification of the recombinant reverse transcriptase is carried out using a combination of the methods described above (3). Single colonies from fresh plates of modified JM109 cells are used to initiate growth of preculture o / n grown at 37 ° C. 2 liters of growth medium is inoculated into the preculture. At OD ₆₀₀ ~ 1.5 (at 37 ℃ 5 to 6 hours) RT induce gene expression with IPTG (1mM final), and continue the fermentation a few hours more at 37 ℃. After centrifugation, the supernatant is discarded while the cell pellets are pooled and stored at -80 ° C until purification. Thaw cells overnight at 4 ° C. and lysis buffer (MES 50 mM pH 6, EDTA 1 mM, 10% v / v glycerol, 0.02% w / v n-octyl-β-D-glucoside, 0.02% w / v sodium azide) Suspend to. Add lysozyme and incubate the mixture for 40 minutes on ice. After homogenization using Dounce in the presence of lysozyme and sonication, the cells are centrifuged for 30 minutes. Collect the supernatant (S1) and store at 4 ℃. Centrifuged pellets were extracted with extraction buffer (MES 50 mM pH 6, KPO ₄ 50 mM pH 6, KCl 100 mM, 10% v / v glycerol, 0.02% w / v n-octyl-β-D-glucoside, 0.02% w / v sodium) Resuspend in azide) and stir at 4 ° C. for 30 minutes. The second mixture is centrifuged again and the supernatant (S2) is collected. The above process is repeated two more times to collect the supernatants S3 and S4, and the last extraction is performed overnight (S5). Polyamine P (0.005% final) is added to the combined supernatant to remove nucleic acid. The solution is stirred at 4 ° C. for 75 minutes and centrifuged for 1 hour. The supernatant (SS1) is precipitated on ice with 20% w / v ammonium sulfate and stirred at 4 ° C. for 1 hour. The mixture is then centrifuged and the resulting supernatant (SS2) is precipitated with additional 40% w / v ammonium sulfate (60% total), stirred for 1 hour and centrifuged again. The final pellet (P1) is stored overnight at 4 ° C. and then purified the next day. Unless stated otherwise, all purification steps are performed at 4 ° C.
[311] Pellets (P1) were reproduced with MES 50 mM pH 6, KPO ₄ 10 mM pH 6, KCl 100 mM, 10% v / v glycerol, 0.02% w / v n-octyl-β-D-glucoside, 0.02% w / v sodium azide Turbid. The suspension is dialyzed overnight against the same buffer using 12-14 kD MWCO dialysis tubing. The dialysate is centrifuged and the supernatant is filtered through a Millex-PF 0.8 μm filter unit. The filtered sample is added to a hydroxyapatite column (30 mL phase volume) and washed with the same buffer. The bound enzyme is eluted with a linear gradient of ˜220 ml of 10-300 mM KPO _{4 in} the above buffer. Fractions containing p66 / p51 heterodimer (as measured by SDS-PAGE 8% and Western blotting) are then pooled for the column. RT containing fractions were diluted 2-fold with bis-tris propane 50 mM pH 7.0, (NH ₄ ) ₂ SO ₄ 1M, 0.02% w / v OBG, 10% v / v glycerol, 0.02% w / v NaN ₃ , Hi-trip heparin Sepharose column (5 mL phase volume) is weighted and washed with the same buffer. The bound RT is then eluted with 75 ml of a linear gradient of 0-1 M ammonium sulfate in the same buffer. RT containing fractions are pooled according to SDS-PAGE and Western blotting analysis. The protein concentration of this pool is measured by the Bradford method using BSA as a standard. The final enzyme preparation is dialyzed in MES 50 mM pH 6, KPO ₄ 300 mM pH 6, KCl 175 mM, 10% v / v glycerol, 0.02% w / v sodium azide, aliquoted and frozen at -80 ° C.
[312] Assay Process:
[313] Radiometric enzyme assays have been applied to the 96-well microtiter plate format and use streptavidin scintillation proximity bead. The assay is briefly described below. Thaw HIV-1 RT enzyme and approximately dilute with Tris / HCl 50 mM pH 7.8 containing 60 mM NaCl, ₂ mM MgCl ₂ hexahydrate, 6 mM DTT, 2 mM reduced glutathione and 0.02% w / v chaps Obtain 3 nM enzyme. To 30 μl of this enzyme solution add 10 μl of inhibitor solution (50 μM to 2.5 nM inhibitor in the same assay buffer as above containing 15% v / v DMSO). The plate is preincubated for 15 minutes at room temperature, then the following steps are performed. In this preculture step, the highest and lowest inhibitor concentrations are 12.5 μM and 0.62 nM, respectively, and the concentration of DMSO is 3.75% v / v. Subsequently, 10 µl of the substrate solution is added to initiate the enzyme reaction. The final reaction mixture was tris / HCl 50 mM pH 7.8, NaCl 60 mM, MgCl ₂ .6H ₂ O 2 mM, DTT 6 mM, GSH 2 mM, Chaps 0.02% w / v DMSO 3% v / v, poly rC 179 nM, biotin dG _{15 18} nM, dGTP 288 nM and ³ H-dGTP 71 nM.
[314] In this culture step, the highest and lowest inhibitor concentrations are 10 μM and 0.5 nM, respectively. After adding the substrate, the plate is covered with a plastic seal and incubated for 1 hour at 37 ° C. in a dry incubator. The reaction is then quenched by adding 75 μl EDTA containing 5 mg / ml of streptavidin scintillation proximity beads. The plate is shaken for 2 minutes at medium speed and incubated for 1 hour at room temperature. 75 μl of cesium chloride 7M solution is then added and the plate is shaken for 2 minutes at medium speed and incubated again for 1 hour at room temperature. The plate is then covered with a plastic seal and TopCount-NXT Count using (Microplate Scintillation & Luminescence Counter, Packard). Each well is counted for 60 seconds. Each column contains blank and control wells at the ends.
[315] % Inhibition is calculated as follows:
[316]
[317] Using the above assay, the present invention is tested for the inhibition of RT wild type (WT) and mutant enzymes. The results are shown in Table 2 as IC ₅₀ (nM).
[318] To confirm the ability of these compounds to inhibit HIV replication, they are also tested in the human T-cell culture (Syncytia) assay described below.
[319] ELISA assay to assay the activity of cell culture
[320] Compounds of the invention are tested for their ability to inhibit HIV replication in cell culture in 96 well plate assays. Complete RPMI 1640, consisting of RPMI 1640 + 10% fetal bovine serum, 10 μg / ml gentamycin and 10 μM β-mercaptoethanol, is used to dilute the compound and at the same time as the cell growth medium. T lymphocyte cell line C8166 is infected with a virus encoding for wild type and mutant reverse transcriptase at a multiplicity of infection of 0.001. The cells are then incubated for three days with the compound of the present invention in the presence of serial dilution. Supernatants were pooled from 8 replicate wells and concentrations of extracellular p24 were available on the market under the HIV-1 p24 antigen assay kit (Beckman-Coulter). Measure with). The level of inhibition (% inhibition) is calculated by the following equation:
[321]
[322] The results are reported in Table 3 as EC ₅₀ (nM).
[323] Specificity test
[324] In order to assay the specificity of the enzymatic inhibitory activity of the compounds provided by the present invention, some of the viruses can be purified using known assays (eg, calf thymus RNA dependent DNA polymerase and human telomerase) as well as other viruses. Test for the ability to inhibit sex polymerases such as hepatitis C virus and respiratory syncytial virus RNA dependent RNA polymerase. It is observed that no compound so tested has any significant inhibitory activity against these enzymes. These results indicate that the enzyme inhibitory activity of the compounds provided by the present invention is specifically for HIV-1 RT.
[325] Literature (incorporated herein by reference)
[326] 1. Benn, S., et al. Science 230: 949, 1985.
[327] 2. D'Aquila, R. T. and Summers, W. C. J. Acq. Imm. Def. Syn. 2: 579, 1989.
[328] 3. a) Warren, T. C. et al. Protein Expression & Purification 3: 479, 1992; b) Kohlstaedt, L. A. Science 256 (5065): 1783, 1992.
[329]
[330]
[331]
[332]

权利要求:
Claims (20)
[1" claim-type="Currently amended] A compound of formula (I) or a pharmaceutically acceptable salt thereof.
Formula I

In Formula I above,
R ² is selected from the group consisting of H, F, Cl, C _1-4 alkyl, C _3-4 cycloalkyl and CF ₃ ,
R ⁴ is H or Me,
R ⁵ is H, Me or Et, R ⁴ and R ⁵ are not both Me, and if R ⁴ is Me, then R ⁵ cannot be Et,
R ¹¹ is Me, Et, cyclopropyl, propyl, isopropyl or cyclobutyl,
Q is selected from the group consisting of Formula (II), (III), (IV) and (V).
Formula II

Formula III

Formula IV

Formula V

[2" claim-type="Currently amended] The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein R ¹¹ is Et or cyclopropyl.
[3" claim-type="Currently amended] The compound of claim 1 wherein Q is A compound selected from the group consisting of or a pharmaceutically acceptable salt thereof.
[4" claim-type="Currently amended] The compound of claim 1 wherein Q is Phosphorus compounds or pharmaceutically acceptable salts thereof.
[5" claim-type="Currently amended] The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R ⁵ is Me.
[6" claim-type="Currently amended] The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R ¹¹ is Et.
[7" claim-type="Currently amended] Chemical formula Of a compound or a pharmaceutically acceptable salt thereof.
[8" claim-type="Currently amended] Chemical formula Of a compound or a pharmaceutically acceptable salt thereof.
[9" claim-type="Currently amended] Chemical formula Of a compound or a pharmaceutically acceptable salt thereof.
[10" claim-type="Currently amended] HIV replication inhibitor of Formula (I) or a pharmaceutically acceptable salt thereof.
Formula I

In Formula I above,
R ² is selected from the group consisting of H, F, Cl, C _1-4 alkyl, C _3-4 cycloalkyl and CF ₃ ,
R ⁴ is H or Me,
R ⁵ is H, Me or Et, R ⁴ and R ⁵ are not both Me, and if R ⁴ is Me, then R ⁵ cannot be Et,
R ¹¹ is Me, Et, cyclopropyl, propyl, isopropyl or cyclobutyl,
Q is selected from the group consisting of Formula (II), (III), (IV) and (V).
Formula II

Formula III

Formula IV

Formula V

[11" claim-type="Currently amended] The HIV replication inhibitor or pharmaceutically acceptable salt thereof according to claim 10, wherein R ¹¹ is Et or cyclopropyl.
[12" claim-type="Currently amended] The compound of claim 10 wherein Q is HIV replication inhibitor or pharmaceutically acceptable salt thereof selected from the group consisting of:
[13" claim-type="Currently amended] The compound of claim 10 wherein Q is HIV replication inhibitors or pharmaceutically acceptable salts thereof.
[14" claim-type="Currently amended] The HIV replication inhibitor of claim 10, wherein R ⁵ is Me.
[15" claim-type="Currently amended] The HIV replication inhibitor of claim 10, wherein R ¹¹ is Et.
[16" claim-type="Currently amended] Chemical formula HIV replication inhibitors or pharmaceutically acceptable salts thereof.
[17" claim-type="Currently amended] Chemical formula HIV replication inhibitors or pharmaceutically acceptable salts thereof.
[18" claim-type="Currently amended] Chemical formula HIV replication inhibitors or pharmaceutically acceptable salts thereof.
[19" claim-type="Currently amended] A method of treating or preventing HIV infection comprising administering to a patient an HIV inhibitory amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1.
[20" claim-type="Currently amended] A pharmaceutical composition for the treatment or prophylaxis of HIV infection comprising the compound of formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

类似技术:

---

公开号 | 公开日 | 专利标题

---

US8658666B2|2014-02-25|Substituted imidazoquinolines and imidazonaphthyridines

---

US8338449B2|2012-12-25|Isoquinolinone potassium channel inhibitors

---

TWI378931B|2012-12-11|Polycyclic carbamoylpyridone derivative having hiv integrase inhibitory activity

---

KR100404166B1|2004-04-28|IMIDAZO[4,5-c]QUINOLINE AMINES

---

KR100286235B1|2001-04-16|Aromatic-linked Polyamine Macrocyclic Compounds Having Anti-HIV Activity

---

KR101773226B1|2017-09-12|Substituted polycyclic carbamoyl pyridone derivative prodrug

---

US7884207B2|2011-02-08|Substituted imidazoquinolines, imidazopyridines, and imidazonaphthyridines

---

US7148237B2|2006-12-12|Nitrogen-containing heteroaryl compounds having HIV integrase inhibitory activity

---

US7968553B2|2011-06-28|Tetrahydro-4H-pyrido[1,2-a] pyrimidines useful as HIV integrase inhibitors

---

CA2413510C|2007-12-11|Condensed pyrazindione derivatives

---

KR100274104B1|2000-12-15|Pyrido[2,3-d]pyrimidine derivatives as phosphodiesterase inhibitors

---

US7968564B2|2011-06-28|HIV integrase inhibitors

---

JP4287649B2|2009-07-01|Aza- and polyaza-naphthalenylcarboxamides useful as HIV integrase inhibitors

---

RU2142463C1|1999-12-10|Tetracyclic derivatives, method of their synthesis |, pharmaceutical composition containing thereof and their use in therapy

---

DE60121461T2|2007-02-08|Condensed pyridoindol derivatives

---

EP1569919B1|2012-04-18|Non-nucleoside reverse transcriptase inhibitors

---

AU2006311871B2|2011-03-03|Hydroxy and alkoxy substituted 1H-imidazoquinolines and methods

---

DK2444400T3|2018-06-18|Substituted polycyclic carbamoylpyridone derivative

---

ES2289159T3|2008-02-01|Replaced tetraciclic derivatives of pirroloquinolona used as phosphodiesterase inhibitors.

---

US8343993B2|2013-01-01|Hydroxyalkyl substituted imidazonaphthyridines

---

US8637671B2|2014-01-28|Indole derivatives as CRTH2 receptor antagonists

---

JP4719151B2|2011-07-06|Isoquinolinone potassium channel inhibitor

---

US5919779A|1999-07-06|5,6-Heteroaryl-dipyrido| azepines and their use in the prevention or treatment of HIV infection

---

US20060128669A1|2006-06-15|Heterocyclic compounds having inhibitory activity against hiv integrase

---

US20080139579A1|2008-06-12|Hiv Integrase Inhibitors

同族专利:

---

公开号 | 公开日

---

MY130375A|2007-06-29|

---

HU0301105A2|2003-08-28|

---

PL360370A1|2004-09-06|

---

IL152819D0|2003-06-24|

---

NZ523549A|2004-08-27|

---

AU2001270370B2|2006-08-03|

---

SA1214B1|2006-09-05|

---

HRP20021003A2|2005-02-28|

---

BG107348A|2004-06-30|

---

ZA200209807B|2003-10-16|

---

DE60118560T2|2006-09-28|

---

CZ20024114A3|2003-04-16|

---

ES2261433T3|2006-11-16|

---

YU94202A|2005-11-28|

---

EA005598B1|2005-04-28|

---

UY26767A1|2002-01-31|

---

CN1178939C|2004-12-08|

---

PT1294720E|2006-06-30|

---

JP2004502787A|2004-01-29|

---

EP1294720B1|2006-04-05|

---

NO20025844L|2002-12-05|

---

TWI270547B|2007-01-11|

---

BR0102377A|2002-02-19|

---

AT322492T|2006-04-15|

---

CA2411766A1|2001-12-20|

---

US20020028807A1|2002-03-07|

---

PE20020083A1|2002-02-28|

---

AR028958A1|2003-05-28|

---

US6420359B1|2002-07-16|

---

DE60118560D1|2006-05-18|

---

NO20025844D0|2002-12-05|

---

EP1294720A1|2003-03-26|

---

DK1294720T3|2006-07-31|

---

AU7037001A|2001-12-24|

---

SK17722002A3|2003-06-03|

---

MXPA02012205A|2003-09-10|

---

CA2411766C|2006-05-23|

---

WO2001096338A1|2001-12-20|

---

HK1057558A1|2005-04-08|

---

NO323830B1|2007-07-09|

---

UA74834C2|2003-04-15|

---

EA200201255A1|2003-06-26|

---

EE200200690A|2004-06-15|

---

CN1436189A|2003-08-13|

---

JP3923894B2|2007-06-06|

引用文献:

---

公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

法律状态:
2000-06-16|Priority to US21232900P

---

2000-06-16|Priority to US60/212,329

---

2000-12-18|Priority to US25663800P

---

2000-12-18|Priority to US60/256,638

---

2001-06-14|Application filed by 베링거 잉겔하임 (캐나다) 리미티드

---

2003-03-03|Publication of KR20030017540A

优先权:

---

申请号 | 申请日 | 专利标题

---

US21232900P| true| 2000-06-16|2000-06-16|

---

US60/212,329|2000-06-16|

---

US25663800P| true| 2000-12-18|2000-12-18|

---

US60/256,638|2000-12-18|