Annelated 9-hydroxy-1,8-dioxo-1,3,4,8-tetrahydro-2H-pyrido[1,2-a]pyrazine-7-carboxamides as HIV integrase inhibitors, methods of preparation and application thereof
20230020329 · 2023-01-19
Inventors
- Alexandre Vasilievich Ivachtchenko (Hallandale, FL, US)
- Andrey Alexandrovich Ivashchenko (Moscow, RU)
- Nikolay Filippovich Savchuk (Rancho Santa Fe, CA)
- Oleg Dmitrievich Mitkin (Khimki, RU)
Cpc classification
A61K9/19
HUMAN NECESSITIES
A61K9/0019
HUMAN NECESSITIES
A61K9/2059
HUMAN NECESSITIES
A61K45/06
HUMAN NECESSITIES
C07D519/00
CHEMISTRY; METALLURGY
International classification
Abstract
The present invention relates to a novel compound that has antiviral activity, in particular, inhibitory activity against the integrase of the human immunodeficiency virus (HIV).
The subject of this invention is a novel annelated 9-hydroxy-1,8-dioxo-1,3,4,8-tetrahydro-2H-pyrido[1,2-a]pyrazine-7-carboxamide of general formula 1 or 2, or any stereoisomer, any pharmaceutically acceptable salt, any solvate, or any crystalline or polycrystalline form thereof
##STR00001## wherein ring A.sup.1 is an optionally methyl-substituted 5-7 membered saturated heterocycle or heterobicycle; ring A.sup.2 is a 5-6 membered optionally methyl-substituted saturated or partially saturated monocyclic heterocycle; ring A.sup.3 is a 5-6 membered monocyclic saturated cycloalkane or tetrahydro-2H-pyran; R is a 5-7 membered optionally substituted with one, two, or three optionally identical substituents monocyclic or bicyclic heterocyclic radical comprising 1-4 heteroatoms selected from the series O, S, and N except (2S,5R,13aS)-8-hydroxy-7,9-dioxo-N-{[3-(trifluoromethyl)-pyridin-2-yl]methyl}-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopurido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide (formula A4) and (1R,4S,12aR)-N-[(3,5-difluoropyridin-2-yl)methyl]-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide (formula A5).
Claims
1. Annelated 9-hydroxy-1,8-dioxo-1,3,4,8-tetrahydro-2H-pyrido[1,2-a]pyrazine-7-carboxamide of general formulas 1 and 2, or its stereoisomer, pharmaceutically acceptable salt, solvate, crystalline or polycrystalline form thereof ##STR00025## wherein ring A.sup.1 is an optionally methyl substituted 5-7 membered saturated heterocycle or heterobicycle, ring A.sup.2 is an optionally methyl substituted 5-6 membered saturated or partially monocyclic heterocycle, ring A.sup.3 is a 5-6 membered monocyclic saturated cycloalkane and tetrahydro-2H-pyran, R is a 5-7-membered, optionally substituted with one, two or three optionally identical substituents, monocyclic or bicyclic heterocyclic radical containing 1-4 heteroatoms selected from the series O, S, and N, except (2S,5R,13aS)-8-hydroxy-7,9-dioxo-N-{[3-(trifluoromethyl)-pyridin-2-yl]methyl}-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopurido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide (formula A4) and (1R,4S,12aR)-N-[(3,5-difluoropyridin-2-yl)methyl]-7-hydroxy-6,8-dioxo-1,2,3,4,6,8,12,12a-octahydro-1,4-methanodipyrido[1,2-a:1′,2′-d]pyrazine-9-carboxamide (formula A5).
2. The compound of claim 1 wherein the heterocyclic radical R is thienyl, furyl, pyrazolyl, isoxazolyl, thiazolyl, oxazolyl, imidazolyl, thiadiazolyl, [1,2,5]oxadiazolyl, [1,2,4]oxadiazolyl, [1,2,4]triazolyl, tetrazolyl, pyridinyl, pyridazinyl, pirimidinyl, pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, and 1,3,5-triazinyl, imidazo[2,1-b]thiazolyl, imidazo[2,1-b][1,3,4]thiadiazolyl, benzothiophenyl, benzofuranyl, indolyl, 1,3-benzodioxol-5-yl, 2,3-dihydro-1,4-benzodioxin-6-yl, 1,3-benzothiazolyl, 1,3-benzoxazolyl, benzimidazolyl, 1,3-dihydro-2trifluorobenzimidazolyl, 2,1,3-benzothiadiazolyl, 2,1,3-benzoxadiazolyl, quinolinyl, isoquinolinyl, imidazo[1,2-a]pyridinyl, 1,2,4-triazolo[4,3-a]pyridinyl, imidazo[1,2-a]pirimidinyl, imidazo[1,2-a]pyrazinyl, 1,2,4-triazolo[4,3-b]pyridazinyl, 4,5,6,7-tetrahydrobenzothiophenyl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyridinyl, 1,4,5,6,7,8-hexahydrocyclohepta[c]pyrazolyl, 5,6,7,8-tetrahydro-4H-cyclohepta[d]thiazolyl, 5,6,7,8-tetrahydro-4H-cyclohepta[d]isothiazol-3-yl, 5,6,7,8-tetrahydro-4H-cyclohepta[d]isothiazol-3-yl, [1,2,4]triazolo[4,3-b]pyridazin-3-yl.
3. The compound of claim 1 or 2 wherein the heterocyclic radical R is 2-thienyl, 2-furyl, 1H-pyrazol-3-yl, 1H-pyrazol-4-yl, 1H-pyrazol-5-yl, isoxazol-4-yl, thiazol-2-yl, 1,3-oxazol-2-yl, imidazol-2-yl, 1,2,3-thiadiazol-5-yl, 1,2,5-oxadiazol-3-yl, 1,2,4-oxadiazol-5-yl, 1H-1,2,4-triazol-3-yl, 1H-1,2,3,4-tetrazol-5-yl, 2-pyridyl, 3-pyridyl, 4-pyridyl, pyridazin-4-yl, pyrimidin-4-yl, pyrazin-2-yl, imidazo[2,1-b]thiazol-6-yl, imidazo[2,1-b][1,3,4]thiadiazol-6-yl, benzothiophen-5-yl, benzofuran-2-yl, 1H-indol-5-yl, 1,3-benzodioxol-5-yl, 2,3-dihydro-1,4-benzodioxin-6-yl, 1,3-benzothiazol-2-yl, 1,3-benzoxazol-2-yl, 1H-benzimidazol-2-yl, 1,3-dihydro-2-oxobenzimidazol-5-yl, 2,1,3-benzothiadiazol-5-yl, 2,1,3-benzoxadiazol-5-yl, quinolin-2-yl, quinolin-3-yl, quinolin-4-yl, quinolin-5-yl, quinolin-6-yl, quinolin-7-yl, quinolin-8-yl, 1-isoquinolinyl, imidazo[1,2-a]pyridin-3-yl, 1,2,4-triazolo[4,3-a]pyridin-3-yl, imidazo[1,2-a]pyrimidin-2-yl, imidazo[1,2-a]pyrazin-3-yl, 1,2,4-triazolo[4,3-b]pyridazin-3-yl, 4,5,6,7-tetrahydrobenzothiophen-2-yl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyridin-3-yl, 1,4,5,6,7,8-hexahydrocyclohepta[c]pyrazol-3-yl, 5,6,7,8-tetrahydro-4H-cyclohepta[d][1,3]thiazol-2-yl, 5,6,7,8-tetrahydro-4H-cyclohepta[d]isothiazol-3-yl, 5,6,7,8-tetrahydro-4H-cyclohepta[d]isothiazol-3-yl, [1,2,4]triazolo[4,3-b]pyridazin-3-yl.
4. The compound according to any one of claims 1, 2, or 3 wherein the heterocyclic radical R is substituted with one, two, or three substituents selected from C1-C3 alkyl or halogen atoms, preferably, F, Cl, and Br.
5. The compound according to any one of claims 1, 2, 3, or 4 of general formula 1.1, 1.2 or 1.3, its stereoisomer, pharmaceutically acceptable salt, solvate, crystalline or polycrystalline form thereof ##STR00026## wherein R is as given above.
6. The compound according to claim 1 of general formula 2.1 or 2.2, its stereoisomer, pharmaceutically acceptable salt, solvate, crystalline or polycrystalline form thereof ##STR00027## wherein R is as given above.
7. The compound according to claim 1 selected from the series: TABLE-US-00006 (3S,11aR)-6-hydroxy-3-methyl-5,7-dioxo-N-(2-thienylmethyl)-2,3,5,7,11,11a- hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.1), (3S,11aR)-N-[(5-bromo-2-thienyl)methyl]-6-hydroxy-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.2), (3S,11aR)-6-hydroxy-3-methyl-N-[(5-methyl-2-furyl)methyl]-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.3), (3S,11aR)-6-hydroxy-N-[(3,5-dimethyl-1H-pyrazol-4-yl)methyl]-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.4), (3S,11aR)-6-hydroxy-3-methyl-N-[(1,3,5-trimethyl-1H-pyrazol-4-yl)methyl]-5,7- dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.5), (3S,11aR)-6-hydroxy-N-[(1-ethyl-3,5-dimethyl-1H-pyrazol-4-yl)methyl]3-methyl-5,7- dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.6), (3S,11aR)-6-hydroxy-N-[(4,5- chloro-1-methyl-1H-pyrazol-3-yl)methyl]3-methyl- 5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.7), (3S,11aR)-6-hydroxy-3-methyl-N-(thiazol-2-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a- hexahydro-[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.8), (3S,11aR)-6-hydroxy-3-methyl-N-[(4-methyl-1,3-oxazol-2-yl)methyl]-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.9), (3S,11aR)-6-hydroxy-3-methyl-N-[(1,4,5-trimethyl-1H-imidazol-2-yl)methyl]-5,7- dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.10), (3S,11aR)-6-hydroxy-3-methyl-N-[(4-methyl-1,2,3-thiadiazol-5-yl)methyl]-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.11), (3S,11aR)-6-hydroxy-3-methyl-N-[(4-methyl-1,2,5-oxadiazol-3-yl)methyl]-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.12), (3S,11aR)-6-hydroxy-3-methyl-N-[(3-methyl-1,2,4-oxadiazol-5-yl)methyl]-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.13), (3S,11aR)-6-hydroxy-3-methyl-N-[(4-methyl-4H-1,2,4-triazol-3-yl)methyl]-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.14), (3S,11aR)-6-hydroxy-3-methyl-N-(pyridyl-2-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a- hexahydro[1,3]-oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.15), (3S,11aR)-6-hydroxy-3-methyl-N-[(3-fluoropyridin-2-yl)methyl]-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.16), (3S,11aR)-6-hydroxy-N-[(3,5-difluoropyridin-2-yl)methyl]-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.17), (3S,11aR)-6-hydroxy-N-[(6-chloropyridin-3-yl)methyl]-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.18), (3S,11aR)-6-hydroxy-N-[(2,6-difluoropyridin-3-yl)methyl]-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.19), Sodium (3S,11aR)-8-({[(2,6-difluoropyridin-3-yl)methyl]amino}carbonyl)-3-methyl- 5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazin-6-olate (1.1.20), (3S,11aR)-6-hydroxy-N-[(2,4-difluoropyridin-3-yl)methyl]-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.21), (3S,11aR)-6-hydroxy-3-methyl-N-[(2,4,6-trifluoropyridin-3-yl)methyl]-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.22), (3S,11aR)-6-hydroxy-3-methyl-N-[(3-fluoropyridin-4-yl)methyl]-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.23), (3S,11aR)-6-hydroxy-N-[(3,5-difluoropyridin-4-yl)methyl]-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.24), (3S,11aR)-6-hydroxy-3-methyl-N-(pyridazin-4-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a- hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.25), (3S,11aR)-6-hydroxy-3-methyl-N-[(2-methylpyrimidin-4-yl)methyl]-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.26), (3S,11aR)-6-hydroxy-3-methyl-N-(pyrazin-2-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a- hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.27), (3S,11aR)-6-hydroxy-3-methyl-N-[(2-chloroimidazo[2,1-b][1,3]thiazol-6-yl)methyl]- 5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.28), (3S,11aR)-6-hydroxy-N-(imidazo[2,1-b][1,3,4]thiadiazol-6-ylmethyl)-3-methyl-5,7- dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.29), (3S,11aR)-6-hydroxy-3-methyl-N-[(2-methylimidazo[2,1-b][1,3,4]thiadiazol-6- yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8- carboxamide (1.1.30), (3S,11aR)-N-(1-benzothien-5-ylmethyl)-6-hydroxy-3-methyl-5,7-dioxo-2,3,5,7,11,11a- hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.31), (3S,11aR)-N-(1-benzofuran-2-ylmethyl)-6-hydroxy-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.32), (3S,11aR)-6-hydroxy-3-methyl-N-[(1-methyl-1H-indol-5-yl)methyl]-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.33), (3S,11aR)-N-(1,3-benzodioxol-5-ylmethyl)-6-hydroxy-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.34), (3S,11aR)-N-[(6-bromo-1,3-benzodioxol-5-yl)methyl]-6-hydroxy-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.35), (3S,11aR)-6-hydroxy-3-methyl-N-[(7-methyl-2,3-dihydro-1,4-benzodioxin-6- yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8- carboxamide (1.1.36), (3S,11aR)-N-(1,3-benzothiazol-2-ylmethyl)-6-hydroxy-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.37), (3S,11aR)-6-hydroxy-3-methyl-N-[(5-chloro-1,3-benzoxazol-2-yl)methyl]-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.38), (3S,11aR)-6-hydroxy-3-methyl-N-[(6-chloro-1,3-benzoxazol-2-yl)methyl]-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.39), (3S,11aR)-6-hydroxy-3-methyl-N-[(5-methyl-1H-benzimidazol-2-yl)methyl]-5,7- dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.40), (3S,11aR)-6-hydroxy-3-methyl-N-[(5-ϕTopo-1H-benzimidazol-2-yl)methyl]-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.41), (3S,11aR)-6-hydroxy-3-methyl-N-[(1-methyl-benzimidazol-2-yl)methyl]-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.42), (3S,11aR)-6-hydroxy-3-methyl-N-[(1-methyl-6-chlorobenzimidazol-2-yl)methyl]-5,7- dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.43), (3S,11aR)-6-hydroxy-3-methyl-N-{[1isopropyl-1H-benzimidazol-2-yl]methyl}-5,7- dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.44), (3S,11aR)-6-hydroxy-3-methyl-N-[(6-chloro-1-isopropyl-1H-benzimidazol-2- yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8- carboxamide (1.1.45), (3S,11aR)-6-hydroxy-N-[(2-oxo-2,3-dihydro-1H-benzimidazol-5-yl)methyl]-3-methyl- 5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.46), (3S,11aR)-N-(2,1,3-benzothiadiazol-5-ylmethyl)-6-hydroxy-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.47), (3S,11aR)-N-(2,1,3-benzoxadiazol-5-ylmethyl)-6-hydroxy-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.48), (3S,11aR)-6-hydroxy-3-methyl-N-(quinolin-2-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a- hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.49), (3S,11aR)-6-hydroxy-3-methyl-N-(quinolin-3-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a- hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.50), (3S,11aR)-6-hydroxy-3-methyl-N-(2-methylquinolin-4-ylmethyl)-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.51), (3S,11aR)-6-hydroxy-3-methyl-N-(quinolin-5-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a- hexahydro[1,3]-oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.52), (3S,11aR)-6-hydroxy-3-methyl-N-(quinolin-6-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a- hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.53), (3S,11aR)-6-hydroxy-3-methyl-N-(quinolin-8-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a- hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.54), (3S,11aR)-6-hydroxy-N-(isoquinolin-1-ylmethyl)-3-methyl-5,7-dioxo-2,3,5,7,11,11a- hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.55), (3S,11aR)-6-hydroxy-N-(imidazo[1,2-a]pyridin-3-ylmethyl)-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.56), (3S,11aR)-6-hydroxy-3-methyl-N-(1,2,4-triazolo[4,3-a]pyridin-3-ylmethyl)-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.57), (3S,11aR)-6-hydroxy-N-(imidazo[1,2-a]pyrimidin-2-ylmethyl)-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.58), (3S,11aR)-6-hydroxy-N-(imidazo[1,2-a]pyrazin-3-ylmethyl)-3-methyl-5,7-dioxo- 2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.59), (3S,11aR)-6-hydroxy-3-methyl-N-[(6-chloro-1,2,4-triazolo[4,3-b]pyridazin-3- yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8- carboxamide (1.1.60), (3S,11aR)-6-hydroxy-3-methyl-N-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyridin-3- ylmethyl)-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8- carboxamide (1.1.61), (3S,11aR)-6-hydroxy-N-(1,4,5,6,7,8-hexahydrocyclohepta[c]pyrazol-3-ylmethyl)-3- methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8- carboxamide (1.1.62), (3S,11aR)-6-hydroxy-3-methyl-5,7-dioxo-N-(5,6,7,8-tetrahydro-4H- cyclohepta[d][1,3]thiazol-2-ylmethyl)-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2- a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.63), (3S,11aR)-6-hydroxy-3-methyl-N-[(1,2-dimethyl-1H-benzimidazol-5-yl)methyl]-5,7- dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.64), (3S,11aR)-6-hydroxy-3-methyl-N-(quinolin-4-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a- hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.65), (4R,12aS)-7-hydroxy-N-[(2,6-difluoropyridin-3-yl)methyl]-4-methyl-6,8-dioxo- 3,4,6,8,12,12a-hexahydro-2H-pyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazine-9-carboxamide (1.2.1), (4R,12aS)-7-hydroxy-N-[(2,4-difluoropyridin-3-yl)methyl]-4-methyl-6,8-dioxo- 3,4,6,8,12,12a-hexahydro-2H-pyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazine-9-carboxamide (1.2.2), (4R,12aS)-7-hydroxy-4-methyl-N-[(2,4.6-trifluoropyridin-3-yl)methyl]-6,8-dioxo- 3,4,6,8,12,12a-hexahydro-2H-pyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazine-9-carboxamide (1.2.3), (4R,12aS)-7-hydroxy-N-[(3,5-difluoropyridin-4-yl)methyl]-4-methyl-6,8-dioxo- 3,4,6,8,12,12a-hexahydro-2H-pyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazine-9-carboxamide (1.2.4), (2R,5S,13aR)-8-hydroxy-N-[(2,6-difluoropyridin-3-yl)methyl]-7,9-dioxo- 2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10- carboxamide (1.3.1), (2R,5S,13aR)-8-hydroxy-N-[(2,4,6-trifluoropyridin-3-yl)methyl]-7,9-dioxo- 2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10- carboxamide (1.3.2), (2R,5S,13aR)-8-hydroxy-N-[(3,5-difluoropyridin-4-yl)methyl]-7,9-dioxo- 2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10- carboxamide (1.3.3), (3aS,6R,13aS)-9-hydroxy-N-[(2,6-difluoropyridin-3-yl)methyl]-6-methyl-8,10-dioxo- 1,2,3,5,6,8,10,13a-octahydrocyclopenta[b][1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-11- carboxamide (2.1.1), (3aS,6R,13aS)-9-hydroxy-N-[(3,5-difluoropyridin-4-yl)methyl]-6-methyl-8,10-dioxo- 1,2,3,5,6,8,10,13a-octahydrocyclopenta[b][1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-11- carboxamide (2.1.2), (3aS,6R,13aS)-9-hydroxy-6-methyl-N-[(2,4,6-trifluoropyridin-3-yl)methyl]-8,10-dioxo- 1,2,3,5,6,8,10,13a-octahydrocyclopenta[b][1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-11- carboxamide (2.1.3), (3aS,6S,13aS)-9-hydroxy-N-[(2,6-difluoropyridin-3-yl)methyl]-6-methyl-8,10-dioxo- 1,2,3,5,6,8,10,13a-octahydrocyclopenta[b][1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-11- carboxamide (2.2.1), (3aS,6S,13aS)-9-hydroxy-N-[(3,5-difluoropyridin-4-yl)methyl]-6-methyl-8,10-dioxo- 1,2,3,5,6,8,10,13a-octahydrocyclopenta[b][1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-11- carboxamide (2.2.2), (3aS,6S,13aS)-9-hydroxy-6-methyl-N-[(2,4,6-trifluoropyridin-3-yl)methyl]-8,10- dioxo-1,2,3,5,6,8,10,13a-octahydrocyclopenta[b][1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-11- carboxamide (2.2.3), or any stereoisomer, any pharmaceutically acceptable salt, any solvate, or any crystalline or polycrystalline form thereof.
8. A method to produce compounds of general formula 1 or 2 according to claim 1 and stereoisomers thereof by the interaction of the corresponding bromides of general formula 3 or 4 and heterocyclylmethylamines in dimethyl sulfoxide in the presence of CO and Pd(PPh.sub.3).sub.4 at elevated temperature followed by debenzylation of the resulting compound 5 or 6 ##STR00028## wherein A.sup.1, A.sup.2, A.sup.3, R, and Bn are as given above.
9. A method to produce compounds of general formula 1 or 2 and stereoisomers thereof by acylation of heterocyclylmethylamines by the corresponding acids of the compounds of general formula 7 or 8 followed by debenzvlation of the resulting compound of formula 5 or 6 ##STR00029## wherein A.sup.1, A.sup.2, A.sup.3, and Bn are as given above.
10. A pharmaceutical composition containing a compound of general formula 1 or 2 according to any one of claims 1-7 in a pharmaceutically effective amount and a pharmaceutically acceptable excipient.
11. The pharmaceutical composition according to claim 10 containing one or more additional therapeutic agents.
12. The pharmaceutical composition according to claim 10 containing an anti-HIV agent as an additional therapeutic agent.
13. The pharmaceutical composition according to claim 12 containing one or more additional therapeutic agents selected from the group consisting of HIV protease inhibitors, HIV non-nucleoside reverse transcriptase inhibitors, HIV nucleoside or nucleotide reverse transcriptase inhibitors, HIV capsid assembly inhibitors and combinations thereof.
14. The pharmaceutical composition according to any one of claims 11-13 containing a first additional therapeutic agent selected from the group consisting of abacavir sulfate, tenofovir, tenofovir disoproxil fumarate, tenofovir alafenamide, tenofovir alafenamide hemifumarate, tenofovir cyclobutylalafenamide, tenofovir cyclobutylalafenamide hemifumarate, and tenofovir cyclobutylalafenamide fumarate, elsulfavirine, and VM-1500A, GS-CA1 and a second additional therapeutic agent selected from the group consisting of emtricitabine and lamivudine.
15. The pharmaceutical composition according to claim 11 containing one or more anti-HBV agents as additional therapeutic agents.
16. The pharmaceutical composition according to claim 11 containing one or more anti-HCV agents as additional therapeutic agents.
17. The pharmaceutical composition according to claim 11 containing one or more anti-HBV agents and one or more anti-HCV agents as additional therapeutic agents.
18. The pharmaceutical composition according to claim 10 in the form of a lyophilizate obtained by freeze drying a nanosuspension of a compound of general formula 1 or 2 in a pharmaceutically effective amount according to any one of claims 1-7 with a particle size of 200 to 900 nm, preferably 200 nm, containing pharmaceutically acceptable excipients.
19. The method to prepare a pharmaceutical composition according to claim 18, said method consisting in grinding wet granules of the compound of general formula 1 or 2 according to any one of claims 1-7 with excipients and water to obtain a particle size of 200 to 900 nm, preferably 200 nm followed by lyophilization of the resulting suspension.
20. The method according to claim 19 wherein excipients are selected from mannite, polysorbate, polyethylene glycol, poloxamer, mannitol, and sucrose.
21. A pharmaceutical nanosuspension to be used as an injectable drug for long-term maintenance therapy of HIV infection containing the pharmaceutical composition according to claim 18, a phosphate buffered saline, and water for injection.
22. A method to prepare a pharmaceutical nanosuspension by mixing the pharmaceutical composition of claim 18, a phosphate buffered saline (PBS) with pH=6.8, and water for injection.
23. An injectable drug for long-term maintenance therapy of HIV infection containing the pharmaceutical composition according to claim 18, a phosphate buffered saline, and water for injection.
24. A method to prepare an injectable drug consisting in mixing the pharmaceutical composition of claim 18, a phosphate buffered saline (PBS) with a pH of 6.8, and water for injection.
25. A method for the prevention and treatment of HIV infection in a HIV-infected or HIV-exposed individual by administering to said individual a therapeutically effective amount of the compound of general formula 1 or 2 according to any one of claims 1-7, or the pharmaceutical composition according to any one of claims 10-18, or the pharmaceutical nanosuspension according to claim 21, or the injectable drug according to claim 23.
Description
[0178] The invention is illustrated by the following drawings:
[0179]
[0180]
[0181]
[0182]
[0183]
BEST EMBODIMENT
[0184] The following examples explain the present invention but are not construed as limiting.
Example 1. Physicochemical Studies of Compounds of General Formula 1 or 2
[0185] General chemical procedures. All chemicals and solvents were used as supplied without further purification. The crude reaction mixtures were concentrated under reduced pressure by removing organic solvents on a rotary evaporator.
[0186] Nuclear Magnetic Resonance Spectra (NMR) were recorded using a Broker DPX-400 spectrometer at room temperature (rt) with tetramethylsilane employed as an internal standard. Chemical shifts (δ) are given in parts per million (ppm), and the signals are presented as s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet) or brs (broad singlet).
[0187] High resolution mass spectra (HRMS) were obtained using an Orbitrap Elite mass spectrometer (Thermo, Bremen, Germany) equipped with a HESI ion source.
[0188] High performance liquid chromatography (HPLC). The purity of the final compounds was determined using HPLC and amounted to more than 98%. The HPLC conditions for purity assessment were as follows: Shimadzu HPLC, XBridge C18, 4.6 mm×250 mm (3.5 μm); a gradient of 0.1% TFA in 5% acetonitrile/water (A) and 0.1% TFA in acetonitrile (B); flow rate 0.5 ml/min; collection time 20 min; and UV wavelengths 214 and 254 nm. The preparative HPLC system included two sets of Shimadzu LC-8A pumps, a Shimadzu SCL 10Avp controller, and a Shimadzu SPD 10Avp detector. A Reprosil-Pur C18-AQ column of 10 μm, 250 mm×20 mm was used. The mobile phase had a gradient of 0.1% TFA in water (A) and 0.1% TFA in acetonitrile (B). LC/MS (LC/MS) was performed on a PE Sciex API 165 system using positive ion electrospray [M+H]+ and a Shimadzu HPLC system equipped with a Waters XBridge C18 3.5 μm column (4.6 mm×150 mm).
[0189] The diastereoisomers were separated on chiral HPLC Phenomenex Lux 5u Cellulose-4, AXIA F, 250×30.00 mm (flow rate: 25 ml/min; UV detector at 215 nm).
[0190] X-ray phase analysis. X-ray diffraction patterns were obtained on a Bruker D8 Advance Vario diffractometer equipped with an X-ray tube with a copper anode and a Ge(III) monochromator (CuKα.sub.1) and a position-sensitive LynxEye detector in the transmission settings. The shooting angle range was 2-60° 2θ and the shooting step, 0.02° 2θ. The analysis was performed using the Bruker Topas 5 software [Bruker TOPAS5 User Manual.—Karlsruhe, Germany: Bruker AXS GmbH, 2015].
Example 2. General Synthetic Procedure for Annelated 9-hydroxy-1,8-dioxo-1,3,4,8-tetrahydro-2H-pyrido[1,2-a]pyrazine-7-carboxamides (General Formula 1 or 2) According to Scheme 1
[0191] Heterocyclylmethylamine or its hydrochloride (0.75 mmol), diisopropylamine (0.131 ml, 0.75 mmol; plus 0.75 mmol per each hydrochloride), and Pd(PPh).sub.4 (29 mg, 0.025 mmol) were added to a solution of corresponding annelated 9-benzyloxy-7-bromo-1,8-dioxo-1,3,4,8-tetrahydro-2H-pyrido[1,2-a]pyrazine (0.5 mmol) of formula 3 or 4 in DMSO (2 ml). The reaction mass was stirred for 14 h at 90° C. under CO. Upon completion of the reaction LC-MS control), the reaction mass was evaporated in vacuum. The residue was dissolved in dichloromethane, washed with water, dried over sodium sulfate, evaporated on a rotary evaporator and subjected to column chromatography on silica gel to yield the corresponding annelated 9-benzyloxy-1,8-dioxo-1,3,4,8-tetrahydro-2H-pyrido[1,2-a]pyrazine-7-carboxamide (formula 6 or 7).
[0192] a) The resulting compound of formula 6 or 7 (0.35 mmol) was dissolved in a mixture of THF (18 ml) and methanol (2 ml), then 10% Pd/C (0.04 g) was added and the mixture was stirred in a hydrogen atmosphere for 8 h. The reaction mass was passed through celite, and the filtrate was evaporated. The residue was treated with ether, the precipitate was filtered off and dried in vacuum to yield the corresponding annelated 9-hydroxy-1,8-dioxo-1,3,4,8-tetrahydro-2H-pyrido[1,2-a]pyrazine-7-carboxamide (general formula 1 or 2).
[0193] b) The compound of formula 6 or 7 (0.35 mmol) was dissolved in 2 ml of trifluoroacetic acid and stirred for 2 h at room temperature. The solution was evaporated in vacuum, the residue was dissolved in chloroform, washed with a saturated NaHCO.sub.3 solution, dried over Na.sub.2SO.sub.4 and evaporated in vacuum. ,
,
, dried in vacuum to yield the corresponding annelated 9-hydroxy-1,8-dioxo-1,3,4,8-tetrahydro-2H-pyrido[1,2-a]pyrazine-7-carboxamide (general formula 1 or 2).
[0194] c) The compound of formula 6 or 7 (0.35 mmol) was dissolved in 1.5 ml of N,N-dimethylacetamide, then 0.148 g (3.5 mmol) of LiCl was added, and the mixture was stirred for 3 h at 80° C. Upon completion of the reaction (LC-MS control), the reaction mass was evaporated in vacuum. The product was isolated by HPLC to yield the corresponding annelated 9-hydroxy-1,8-dioxo-1,3,4,8-tetrahydro-2H-pyrido[1,2-a]pyrazine-7-carboxamide (general formula 1 or 2) including
[0195] (3S,11aR)-6-hydroxy-3-methyl-5,7-dioxo-N-(2-thienylmethyl)-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.1): LC-MS (M+1)=376; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.47 (brs, 1H), 10.33 (t, J=4.8 Hz, 1H), 8.49 (s, 1H), 7.40 (dd, J.sub.1=8.8 Hz, J.sub.2=4.8 Hz, 1H), 7.03 (d, J=2.8 Hz, 1H), 6.96 (dd, J.sub.1=4.8 Hz, J.sub.2=3.6 Hz, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.90 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.70 (d, J=5.6 Hz, 2H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 4.29 (m, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.67 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 1.34 (d, J=6.0 Hz, 3H);
[0196] (3S,11aR)-6-hydroxy-3-methyl-N-[(5-methyl-2-furyl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.3): LC-MS (ESI) 374 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.46 (brs, 1H), 10.20 (t, J=5.2 Hz, 1H), 8.46 (s, 1H), 6.15 (d, J=2.4 Hz, 1H), 5.99 (d, J=2.4 Hz, 1H), 5.39 (dd, J.sub.1=9.6 Hz, J.sub.2=4.0 Hz, 1H), 4.89 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.46 (d, J=5.2 Hz, 2H), 4.39 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 4.29 (m, 1H), 4.00 (t, J=11.0 Hz, 1H), 3.66 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 2.23 (s, 3H), 1.34 (d, J=6.0 Hz, 3H);
[0197] (3S,11aR)-6-hydroxy-N-[(3,5-dimethyl-1H-pyrazol-4-yl)methyl]-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.4). LC-MS (ESI) 388 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.04 (brs, 1H), 11.42 (brs, 1H), 9.92 (t, J=4.8 Hz, 1H), 8.46 (s, 1H), 5.38 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.88 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.38 (dd, J.sub.1=8.0 Hz, J.sub.2=7.2 Hz, 1H), 4.29 (m, 1H), 4.26 (d, J=5.2 Hz, 2H), 4.00 (t, J=11.2 Hz, 1H), 3.66 (dd, J.sub.1=8.0 Hz, J.sub.2=7.2 Hz, 1H), 1.33 (d, J=6.4 Hz, 3H);
[0198] (3S,11aR)-6-hydroxy-3-methyl-N-[(1,3,5-trimethyl-1H-pyrazol-4-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.5): LC-MS (ESI) 402 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.40 (brs, 1H), 9.92 (t, J=4.8 Hz, 1H), 8.46 (s, 1H), 5.38 (dd, J.sub.1=9.6 Hz, J.sub.2=3.6 Hz, 1H), 4.88 (dd, J.sub.1=12.0 Hz, J.sub.2=3.6 Hz, 1H), 4.39 (t, J=7.4 Hz, 1H), 4.29 (m, 1H), 4.25 (d, J=4.8 Hz, 2H), 4.00 (t, J=11.0 Hz, 1H), 3.66 (t, J=7.4 Hz, 1H), 3.32 (s, 3H), 2.19 (s, 3H), 2.08 (s, 3H), 1.33 (d, J=6.0 Hz, 3H);
[0199] (3S,11aR)-6-hydroxy-N-[(1-ethyl-3,5-dimethyl-1H-pyrazol-4-yl)methyl]3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.6): LC-MS (ESI) 416 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.40 (brs, 1H), 9.92 (t, J=5.2 Hz, 1H), 8.47 (s, 1H), 5.38 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.88 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.39 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 4.28 (m, 1H), 4.25 (d, J=5.6 Hz, 2H), 4.00 (t, J=11.2 Hz, 1H), 3.94 (q, J=7.2 Hz, 2H), 3.66 (dd, J.sub.1=8.4 Hz, J.sub.2=6.4 Hz, 1H), 2.20 (s, 3H), 2.09 (s, 3H), 1.33 (d, J=6.0 Hz, 3H), 1.24 (t, J=7.2 Hz, 3H);
[0200] (3S,11aR)-6-hydroxy-N-[(4,5-∂uchloro-1-methyl-1H-pyrazol-3-yl)methyl]3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.7). LC-MS (ESI) 443 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.44 (brs, 1H), 10.28 (t, J=5.6 Hz, 1H), 8.46 (s, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 4.89 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.50 (d, J=5.6 Hz, 2H), 4.39 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 4.29 (sxt, J=6.4 Hz, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.80 (s, 3H), 3.66 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 1.34 (d, J=6.0 Hz, 3H);
[0201] (3S,11aR)-6-hydroxy-3-methyl-N-(thiazol-2-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.8): LC-MS (ESI) 377 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.50 (brs, 1H), 10.58 (t, J=6.0 Hz, 1H), 8.49 (s, 1H), 7.74 (d, J=3.2 Hz, 1H), 7.62 (d, J=3.2 Hz, 1H), 5.40 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.90 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.84 (d, J=6.0 Hz, 2H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 4.30 (m, 1H), 4.01 (dd, J.sub.1=12.0 Hz, J.sub.2=10.0 Hz, 1H), 3.67 (dd, J.sub.1=8.4 Hz, J.sub.2=6.4 Hz, 1H), 1.35 (d, J=6.4 Hz, 3H);
[0202] (3S,11aR)-6-hydroxy-3-methyl-N-[(4-methyl-1,2,5-oxadiazol-3-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.12): LC-MS (ESI) 376 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.51 (brs, 1H), 10.45 (t, J=5.6 Hz, 1H), 8.47 (s, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.89 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.74 (d, J=5.6 Hz, 2H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 4.29 (m, 1H), 4.00 (t, J=11.0 Hz, 1H), 3.67 (dd, J.sub.1=8.0 Hz, J.sub.2=7.2 Hz, 1H), 2.37 (s, 3H), 1.34 (d, J=6.0 Hz, 3H);
[0203] (3S,11aR)-6-hydroxy-3-methyl-N-[(4-methyl-4H-1,2,4-triazol-3-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.14). LC-MS (ESI) 375 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.49 (brs, 1H), 10.41 (t, J=5.2 Hz, 1H), 8.48 (s, 1H), 8.40 (s, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 4.89 (dd, J.sub.1=11.6 Hz, J.sub.2=3.6 Hz, 1H), 4.69 (d, J=5.2 Hz, 2H), 4.39 (t, J=7.6 Hz, 1H), 4.29 (m, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.66 (m, 1H), 3.65 (s, 3H), 1.34 (d, J=6.0 Hz, 3H);
[0204] (3S,11aR)-6-hydroxy-3-methyl-N-(pyridyl-2-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.15): LC-MS (ESI) 371 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.44 (brs, 1H), 10.52 (t, J=5.6 Hz, 1H), 8.53 (d, J=4.0 Hz, 1H), 8.48 (s, 1H), 7.76 (t, J=7.6 Hz, 1H), 7.33 (d, J=8.0 Hz, 1H), 7.28 (m, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 4.89 (dd, J.sub.1=12.0 Hz, J.sub.2=3.6 Hz, 1H), 4.64 (d, J=5.6 Hz, 2H), 4.40 (t, J=7.6 Hz, 1H), 4.30 (m, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.66 (dd, J.sub.1=8.0 Hz, J.sub.2=6.8 Hz, 1H), 1.34 (d, J=6.0 Hz, 3H);
[0205] (3S,11aR)-6-hydroxy-3-methyl-N-[(2-methylpyrimidin-4-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.26): LC-MS (M+1)=386; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.50 (brs, 1H), 10.50 (t, J=6.0 Hz, 1H), 8.61 (d, J=5.0 Hz, 1H), 8.46 (s, 1H), 7.16 (d, J=5.0 Hz, 1H), 5.40 (dd, J.sub.1=10.0 Hz, J.sub.2=3.2 Hz, 1H), 4.89 (dd, J.sub.1=12.0 Hz, J.sub.2=3.2 Hz, 1H), 4.59 (d, J=6.0 Hz, 2H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 4.30 (m, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.67 (dd, J.sub.1=8.4 Hz, J.sub.2=6.4 Hz, 1H), 2.60 (s, 3H), 1.35 (d, J=6.0 Hz, 3H);
[0206] (3S,11aR)-N-(1-benzothien-5-ylmethyl)-6-hydroxy-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.31): LC-MS (ESI) 426 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.47 (brs, 1H), 10.38 (t, J=6.0 Hz, 1H), 8.50 (s, 1H), 7.96 (d, J=8.4 Hz, 1H), 7.80 (s, 1H), 7.76 (d, J=9.4 Hz, 1H), 7.43 (d, J=9.4 Hz, 1H), 7.33 (dd, J.sub.1=8.4 Hz, J.sub.2=0.6 Hz, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.90 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.66 (d, J=5.6 Hz, 2H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 4.30 (m, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.67 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 1.34 (d, J=6.0 Hz, 3H).
[0207] (3S,11aR)-N-(1-benzofuran-2-ylmethyl)-6-hydroxy-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.32): LC-MS (ESI) 410 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.50 (brs, 1H), 10.41 (t, J=5.6 Hz, 1H), 8.50 (s, 1H), 7.59 (d, J=7.6 Hz, 1H), 7.54 (d, J=7.6 Hz, 1H), 7.27 (t, J=7.6 Hz, 1H), 7.22 (t, J=7.2 Hz, 1H), 6.75 (s, 1H), 5.40 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 4.91 (dd, J.sub.1=12.0 Hz, J.sub.2=3.6 Hz, 1H), 4.72 (d, J=5.6 Hz, 2H), 4.40 (t, J=7.6 Hz, 1H), 4.28 (sxt, J=6.0 Hz, 1H), 4.02 (t, J=11.0 Hz, 1H), 3.67 (t, J=7.6 Hz, 1H), 1.34 (d, J=6.0 Hz, 3H).
[0208] (3S,11aR)-6-hydroxy-3-methyl-N-[(1-methyl-1H-indol-5-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.33): LC-MS (ESI) 423 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.43 (brs, 1H), 10.27 (t, J=6.0 Hz, 1H), 8.50 (s, 1H), 7.48 (s, 1H), 7.39 (d, J=8.8 Hz, 1H), 7.30 (d, J=2.8 Hz, 1H), 7.12 (dd, J.sub.1=8.8 Hz, J.sub.2=0.8 Hz, 1H), 6.37 (dd, J.sub.1=2.8 Hz, J.sub.2=0.4 Hz, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.90 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.59 (d, J=5.6 Hz, 2H), 4.39 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 4.29 (m, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.77 (s, 3H), 3.66 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 1.34 (d, J=6.0 Hz, 3H).
[0209] (3S,11aR)-N-(1,3-benzodioxol-5-ylmethyl)-6-hydroxy-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.34): LC-MS (ESI) 414 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.45 (brs, 1H), 10.24 (t, J=5.8 Hz, 1H), 8.48 (s, 1H), 6.86 (m, 2H), 6.79 (m, 1H), 5.98 (s, 2H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.89 (dd, J.sub.1=12.4 Hz, J.sub.2=4.0 Hz, 1H), 4.43 (d, J=6.0 Hz, 2H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 4.29 (m, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.67 (dd, J.sub.1=8.4 Hz, J.sub.2=6.0 Hz, 1H), 1.34 (d, J=6.4 Hz, 3H).
[0210] (3S,11aR)-N-[(6-bromo-1,3-benzodioxol-5-yl)methyl]-6-hydroxy-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.35): LC-MS (ESI) 493 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.44 (brs, 1H), 10.30 (m, 1H), 8.46 (d, J=2.4 Hz, 1H), 7.23 (d, J=2.4 Hz, 1H), 6.93 (d, J=2.4 Hz, 1H), 6.05 (d, J=2.4 Hz, 2H), 5.38 (m, 1H), 4.88 (m, 1H), 4.46 (m, 2H), 4.39 (m, 1H), 4.29 (m, 1H), 4.00 (m, 1H), 3.66 (m, 1H), 1.34 (dd, J.sub.1=6.4 Hz, J.sub.2=2.8 Hz, 3H).
[0211] (3S,11aR)-6-hydroxy-3-methyl-N-[(7-methyl-2,3-dihydro-1,4-benzodioxin-6-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.36). LC-MS (ESI) 434 (M+H); .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.43 (brs, 1H), 10.14 (t, J=5.6 Hz, 1H), 8.47 (s, 1H), 6.73 (s, 1H), 6.68 (s, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.89 (dd, J.sub.1=12.0 Hz, J.sub.2=3.6 Hz, 1H), 4.39 (m, 3H), 4.29 (m, 1H), 4.18 (s, 4H), 4.00 (t, J=11.2 Hz, 1H), 3.66 (dd, J.sub.1=8.0 Hz, J.sub.2=7.2 Hz, 1H), 2.17 (s, 3H), 1.34 (d, J=6.0 Hz, 3H).
[0212] (3S,11aR)-N-(1,3-benzothiazol-2-ylmethyl)-6-hydroxy-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.37): LC-MS (ESI) 427 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.54 (brs, 1H), 10.69 (t, J=5.6 Hz, 1H), 8.50 (s, 1H), 8.04 (d, J=8.0 Hz, 1H), 7.95 (d, J=8.0 Hz, 1H), 7.50 (t, J=7.2 Hz, 1H), 7.41 (t, J=7.2 Hz, 1H), 5.40 (dd, J.sub.1=9.6 Hz, J.sub.2=3.6 Hz, 1H), 4.97 (d, J=5.6 Hz, 2H), 4.90 (dd, J.sub.1=12.0 Hz, J.sub.2=3.6 Hz, 1H), 4.40 (t, J=7.6 Hz, 1H), 4.30 (m, 1H), 4.02 (t, J=11.2 Hz, 1H), 3.67 (t, J=7.6 Hz, 1H), 1.35 (d, J=6.0 Hz, 3H).
[0213] (3S,1aR)-6-hydroxy-3-methyl-N-[(5-chloro-1,3-benzoxazol-2-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.38): LC-MS (ESI) 445 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.52 (brs, 1H), 10.54 (t, J=5.6 Hz, 1H), 8.47 (s, 1H), 7.83 (d, J=2.0 Hz, 1H), 7.75 (d, J=8.8 Hz, 1H), 7.42 (dd, J.sub.1=8.8 Hz, J.sub.2=2.0 Hz, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.88 (m, 3H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 4.30 (m, 1H), 4.01 (dd, J.sub.1=11.6 Hz, J.sub.2=6.4 Hz, 1H), 3.67 (dd, J.sub.1=8.4 Hz, J.sub.2=6.4 Hz, 1H), 1.35 (d, J=6.4 Hz, 3H).
[0214] (3S,11aR)-6-hydroxy-3-methyl-N-[(6-chloro-1,3-benzoxazol-2-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.39): LC-MS (ESI) 445 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.52 (brs, 1H), 10.54 (t, J=5.4 Hz, 1H), 8.47 (s, 1H), 7.92 (d, J=1.2 Hz, 1H), 7.72 (d, J=8.4 Hz, 1H), 7.41 (dd, J.sub.1=8.4 Hz, J.sub.2=1.6 Hz, 1H), 5.40 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.89 (m, 1H), 4.87 (d, J=6.0 Hz, 2H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 4.30 (m, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.67 (dd, J.sub.1=8.0 Hz, J.sub.2=6.8 Hz, 1H), 1.35 (d, J=6.4 Hz, 3H).
[0215] (3S,11aR)-6-hydroxy-3-methyl-N-[(5-methyl-1H-benzimidazol-2-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.40): LC-MS (ESI) 424 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.19 (s, 1H), 11.47 (brs, 1H), 10.48 (s, 1H), 8.47 (s, 1H), 7.32 (m, 2H), 6.96 (m, 1H), 5.40 (m, 1H), 4.90 (m, 1H), 4.72 (m, 2H), 4.40 (m, 1H), 4.30 (m, 1H), 4.02 (m, 1H), 3.67 (m, 1H), 2.39 (s, 3H), 1.35 (d, J=4.4 Hz, 3H).
[0216] (3S,11aR)-6-hydroxy-3-methyl-N-[(5-ϕmopo-H-benzimidazol-2-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.41): LC-MS (ESI) 428 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.49 (brs, 1H), 10.51 (brs, 1H), 8.47 (s, 1H), 7.52 (m, 1H), 7.33 (d, J=8.4 Hz, 1H), 7.03 (t, J=8.4 Hz, 1H), 5.40 (m, 1H), 4.90 (m, 1H), 4.77 (brs, 2H), 4.40 (m, 1H), 4.30 (m, 1H), 4.02 (t, J=10.8 Hz, 1H), 3.67 (m, 1H), 1.35 (d, J=5.6 Hz, 3H).
[0217] (3S,11aR)-6-hydroxy-3-methyl-N-[(1-methyl-6-chlorobenzimidazol-2-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.43): LC-MS (ESI) 458 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.47 (brs, 1H), 10.58 (t, J=5.2 Hz, 1H), 8.50 (s, 1H), 7.65 (d, J=1.2 Hz, 1H), 7.58 (d, J=8.4 Hz, 1H), 7.27 (dd, J.sub.1=8.4 Hz, J.sub.2=1.2 Hz, 1H), 5.40 (dd, J.sub.1=9.6 Hz, J.sub.2=3.2 Hz, 1H), 4.90 (m, 1H), 4.85 (d, J=5.2 Hz, 2H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 4.30 (m, 1H), 4.02 (t, J=11.0 Hz, 1H), 3.81 (s, 3H), 3.67 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 1.34 (d, J=6.0 Hz, 3H).
[0218] (3S,11aR)-6-hydroxy-3-methyl-N-{[1-isopropyl-]H-benzimidazol-2-yl]methyl}-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.44): LC-MS (ESI) 452 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.47 (brs, 1H), 10.57 (t, J=5.2 Hz, 1H), 8.52 (s, 1H), 7.70 (d, J=8.0 Hz, 1H), 7.61 (d, J=8.0 Hz, 1H), 7.18 (m, 2H), 5.40 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.91 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.86 (d, J=5.2 Hz, 2H), 4.83 (m, 1H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 4.29 (m, 1H), 4.02 (dd, J.sub.1=12.0 Hz, J.sub.2=10.0 Hz, 1H), 3.66 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 1.56 (d, J=6.8 Hz, 6H), 1.34 (d, J=6.0 Hz, 3H).
[0219] (3S,11aR)-6-hydroxy-N-[(2-oxo-2,3-dihydro-1H-benzimidazol-5-yl)methyl]-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.46). LC-MS (ESI) 426 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.46 (brs, 1H), 10.56 (s, 2H), 10.26 (t, J=5.6 Hz, 1H), 8.48 (s, 1H), 6.88 (m, 3H), 5.39 (m, 1H), 4.90 (m, 1H), 4.49 (d, J=5.6 Hz, 2H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 4.29 (m, 1H), 4.01 (t, J=11.0 Hz, 1H), 3.66 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 1.34 (d, J=6.0 Hz, 3H).
[0220] (3S,11aR)-N-(2,1,3-benzoxadiazol-5-yl)methyl)-6-hydroxy-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.47): LC-MS (ESI) 412 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.51 (brs, 1H), 10.46 (t, J=6.0 Hz, 1H), 8.49 (s, 1H), 8.03 (d, J=9.4 Hz, 1H), 7.78 (s, 1H), 7.57 (d, J=9.4 Hz, 1H), 5.40 (dd, J.sub.1=9.8 Hz, J.sub.2=3.4 Hz, 1H), 4.90 (dd, J.sub.1=12.0 Hz, J.sub.2=3.4 Hz, 1H), 4.68 (d, J=5.6 Hz, 2H), 4.40 (t, J=7.6 Hz, 1H), 4.30 (m, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.67 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 1.35 (d, J=6.0 Hz, 3H).
[0221] (3S,11aR)-6-hydroxy-3-methyl-N-(quinolin-2-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]-oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.49): LC-MS (ESI) 421 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.44 (brs, 1H), 10.30 (t, J=5.6 Hz, 1H), 8.50 (s, 1H), 7.45 (s, 1H), 7.42 (d, J=8.0 Hz, 1H), 7.16 (dd, J.sub.1=8.0 Hz, J.sub.2=0.8 Hz, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 4.90 (m, 1H), 4.61 (d, J=5.6 Hz, 2H), 4.39 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 4.29 (sxt, J=6.4 Hz, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.70 (s, 3H), 3.66 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 2.50 (s, 3H), 1.34 (d, J=6.4 Hz, 3H).
[0222] (3S,11aR)-6-hydroxy-3-methyl-N-(quinolin-3-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.50): LC-MS (ESI) 421 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.50 (brs, 1H), 10.47 (t, J=6.0 Hz, 1H), 8.91 (d, J=2.0 Hz, 1H), 8.50 (s, 1H), 8.24 (s, 1H), 8.02 (d, J=8.0 Hz, 1H), 7.96 (d, J=7.6 Hz, 1H), 7.74 (dt, J.sub.1=8.0 Hz, J.sub.2=1.2 Hz, 1H), 7.61 (m, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.89 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.76 (d, J=6.0 Hz, 2H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 4.30 (m, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.67 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 1.35 (d, J=6.0 Hz, 3H).
[0223] (3S,11aR)-6-hydroxy-3-methyl-N-(2-methylquinolin-4-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.51): LC-MS (ESI) 435 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.49 (brs, 1H), 10.47 (t, J=5.6 Hz, 1H), 8.52 (s, 1H), 8.14 (d, J=8.4 Hz, 1H), 7.95 (d, J=8.4 Hz, 1H), 7.73 (t, J=7.6 Hz, 1H), 7.57 (t, J=7.2 Hz, 1H), 7.28 (s, 1H), 5.40 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 5.03 (d, J=5.6 Hz, 2H), 4.91 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 4.30 (m, 1H), 4.02 (t, J=11.2 Hz, 1H), 3.67 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 2.61 (s, 3H), 1.34 (d, J=6.4 Hz, 3H).
[0224] (3S,11aR)-6-hydroxy-3-methyl-N-(quinolin-5-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.52): LC-MS (ESI) 421 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.44 (s, 1H), 10.42 (t, J=6.0 Hz, 1H), 8.92 (dd, J.sub.1=8.0 Hz, J.sub.2=1.2 Hz, 1H), 8.61 (d, J=8.0 Hz, 1H), 8.52 (s, 1H), 7.97 (d, J=8.4 Hz, 1H), 7.73 (t, J=8.2 Hz, 1H), 7.58 (m, 2H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 5.03 (d, J=6.0 Hz, 2H), 4.91 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.39 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 4.29 (sxt, J=6.4 Hz, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.66 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 1.33 (d, J=6
[0225] (3S,11aR)-6-hydroxy-3-methyl-N-(quinolin-8-ylmethyl)-5,7-dioxo-2,3,5,7,11,1a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.54): LC-MS (ESI) 421 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.41 (brs, 1H), 10.49 (t, J=5.6 Hz, 1H), 8.99 (dd, J.sub.1=4.0 Hz, J.sub.2=1.6 Hz, 1H), 8.47 (s, 1H), 8.40 (dd, J.sub.1=8.0 Hz, J.sub.2=1.2 Hz, 1H), 7.91 (d, J=8.0 Hz, 1H), 7.67 (d, J=6.8 Hz, 1H), 7.58 (m, 2H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 5.13 (d, J=5.6 Hz, 2H), 4.88 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.39 (dd, J.sub.1=8.0 Hz, J.sub.2=7.6 Hz, 1H), 4.29 (m, 1H), 4.00 (t, J=11.2 Hz, 1H), 3.66 (dd, J.sub.1=8.0 Hz, J.sub.2=6.8 Hz, 1H), 1.34 (d, J=6.0 Hz, 3H).
[0226] (3S,11aR)-6-hydroxy-3-methyl-N-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyridin-3-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.61): LC-MS (ESI) 415 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.48 (brs, 1H), 10.38 (t, J=5.6 Hz, 1H), 8.48 (s, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 4.89 (dd, J.sub.1=12.0 Hz, J.sub.2=3.6 Hz, 1H), 4.64 (d, J=5.6 Hz, 2H), 4.39 (dd, J.sub.1=8.0 Hz, J.sub.2=7.2 Hz, 1H), 4.29 (sxt, J=6.0 Hz, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.93 (t, J=6.0 Hz, 2H), 3.66 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 2.79 (t, J=6.4 Hz, 2H), 1.88 (m, 2H), 1.79 (m, 2H), 1.34 (d, J=6.0 Hz, 3H).
[0227] (3S,11aR)-6-hydroxy-N-(1,4,5,6,7,8-hexahydrocyclohepta[c]pyrazol-3-ylmethyl)-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide 1.1.62: LC-MS (ESI) 428 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.13 (brs, 1H), 11.43 (brs, 1H), 10.07 (t, J=5.2 Hz, 1H), 8.47 (s, 1H), 5.40 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 4.89 (dd, J.sub.1=12.0 Hz, J.sub.2=3.6 Hz, 1H), 4.39 (m, 3H), 4.29 (m, 1H), 4.00 (t, J=11.2 Hz, 1H), 3.66 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 2.63 (m, 2H), 2.45 (m, 2H), 1.75 (m, 2H), 1.55 (m, 4H), 1.34 (d, J=6.0 Hz, 3H).
[0228] (3S,11aR)-6-hydroxy-3-methyl-5,7-dioxo-N-(5,6,7,8-tetrahydro-4H-cyclohepta[d][1,3]thiazol-2-ylmethyl)-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.63): LC-MS (ESI) 445 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.49 (brs, 1H), 10.47 (m, 1H), 8.48 (s, 1H), 5.39 (m, 1H), 4.90 (m, 1H), 4.67 (m, 2H), 4.40 (m, 1H), 4.30 (m, 1H), 4.01 (m, 1H), 3.67 (m, 1H), 2.82 (m, 2H), 2.73 (m, 2H), 1.78 (m, 2H), 1.60 (m, 4H), 1.34 (dd, J.sub.1=6.4 Hz, J.sub.2=2.8 Hz, 3H).
[0229] (3S,11aR)-6-hydroxy-3-methyl-N-[(1,2-dimethyl-1H-benzimidazol-5-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.64): LC-MS (ESI) 438 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.44 (brs, 1H), 10.30 (t, J=5.6 Hz, 1H), 8.50 (s, 1H), 7.45 (s, 1H), 7.42 (d, J=8.0 Hz, 1H), 7.16 (dd, J.sub.1=8.0 Hz, J.sub.2=0.8 Hz, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 4.90 (m, 1H), 4.61 (d, J=5.6 Hz, 2H), 4.39 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 4.29 (sxt, J=6.4 Hz, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.70 (s, 3H), 3.66 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 2.50 (s, 3H), 1.34 (d, J=6.4 Hz, 3H).
[0230] (3S,11aR)-6-hydroxy-3-methyl-N-(quinolin-4-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.65): LC-MS (ESI) 421 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.49 (brs, 1H), 10.50 (t, J=6.0 Hz, 1H), 8.85 (d, J=4.0 Hz, 1H), 8.52 (s, 1H), 8.21 (d, J=8.8 Hz, 1H), 8.06 (d, J=8.0 Hz, 1H), 7.79 (t, J=6.8 Hz, 1H), 7.66 (t, J=7.6 Hz, 1H), 7.39 (d, J=4.0 Hz, 1H), 5.40 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 5.08 (d, J=6.0 Hz, 2H), 4.91 (dd, J.sub.1=12.0 Hz, J.sub.2=3.6 Hz, 1H), 4.40 (dd, J.sub.1=8.0 Hz, J.sub.2=7.2 Hz, 1H), 4.29 (m, 1H), 4.02 (t, J=11.2 Hz, 1H), 3.67 (dd, J.sub.1=8.0 Hz, J.sub.2=6.8 Hz, 1H), 1.33 (d, J=6.0 Hz, 3H).
[0231] (3aS,6R,13aS)-9-hydroxy-N-[(2,6-difluoropyridin-3-yl)methyl]-6-methyl-8,10-dioxo-1,2,3,5,6,8,10,13a-octahydrocyclopenta[b][1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-11-carboxamide (2.1.1): LC-MS (ESI) 447 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.28 (s, 1H), 10.39 (t, J=6.0 Hz, 1H), 8.41 (s, 1H), 8.08 (q, J=8.4 Hz, 1H), 7.14 (dd, J.sub.1=8.4 Hz, J.sub.2=2.0 Hz, 1H), 4.55 (m, 2H), 4.36 (m, 3H), 3.76 (dd, J.sub.1=8.0 Hz, J.sub.2=6.0 Hz, 1H), 2.33 (m, 1H), 2.14 (m, 1H), 1.77 (m, 3H), 1.38 (d, J=6.0 Hz, 3H).
[0232] (3aS,6R,13aS)-9-hydroxy-N-[(3,5-difluoropyridin-4-yl)methyl]-6-methyl-8,10-dioxo-1,2,3,5,6,8,10,13a-octahydrocyclopenta[b]1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-11-carboxamide (2.1.2): LC-MS (ESI) 447 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.28 (s, 1H), 10.48 (t, J=6.0 Hz, 1H), 8.50 (s, 2H), 8.38 (s, 1H), 4.66 (m, 2H), 4.34 (m, 3H), 3.75 (dd, J.sub.1=8.0 Hz, J.sub.2=6.0 Hz, 1H), 2.30 (m, 1H), 2.12 (m, 1H), 1.76 (m, 3H), 1.36 (d, J=6.0 Hz, 3H).
[0233] (3aS,6R,13aS)-9-hydroxy-6-methyl-N-[(2,4,6-trifluoropyridin-3-yl)methyl]-8,10-dioxo-1,2,3,5,6,8,10,13a-octahydrocyclopenta[b][1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-11-carboxamide (2.1.3): LC-MS (ESI) 465 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.28 (s, 1H), 10.42 (t, J=5.6 Hz, 1H), 8.38 (s, 1H), 7.34 (d, J=8.0 Hz, 1H), 4.54 (m, 2H), 4.35 (m, 3H), 3.75 (m, 1H), 2.31 (m, 1H), 2.12 (m, 1H), 1.76 (m, 3H), 1.36 (d, J=6.0 Hz, 3H).
[0234] (3aS,6S,13aS)-9-hydroxy-N-[(2,6-difluoropyridin-3-yl)methyl]-6-methyl-8,10-dioxo-1,2,3,5,6,8,10,13a-octahydrocyclopenta[b][1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-11-carboxamide (2.2.1): LC-MS (ESI) 447 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.29 (s, 1H), 10.40 (t, J=5.8 Hz, 1H), 8.41 (s, 1H), 8.08 (q, J=8.4 Hz, 1H), 7.15 (dd, J.sub.1=8.4 Hz, J.sub.2=2.0 Hz, 1H), 4.55 (m, 2H), 4.35 (m, 3H), 3.75 (dd, J.sub.1=8.0 Hz, J.sub.2=6.4 Hz, 1H), 2.33 (m, 1H), 2.13 (m, 1H), 1.76 (m, 3H), 1.37 (d, J=6.4 Hz, 3H).
[0235] (3aS,6S,13aS)-9-hydroxy-N-[(3,5-difluoropyridin-4-yl)methyl]-6-methyl-8,10-dioxo-1,2,3,5,6,8,10,13a-octahydrocyclopenta[b][1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-11-carboxamide (2.2.2): LC-MS (ESI) 447 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.46 (s, 1H), 10.47 (t, J=5.8 Hz, 1H), 8.49 (s, 2H), 8.42 (s, 1H), 5.43 (dd, J.sub.1=9.2 Hz, J.sub.2=4.0 Hz, 1H), 5.09 (brs, 1H), 4.66 (m, 3H), 4.60 (brs, 1H), 4.01 (dd, J.sub.1=12.4 Hz, J.sub.2=9.6 Hz, 1H), 1.94 (brs, 4H), 1.84 (d, J=12.0 Hz, 1H), 1.57 (m, 1H).
[0236] (3aS,6S,13aS)-9-hydroxy-6-methyl-N-[(2,4,6-trifluoropyridin-3-yl)methyl]-8,10-dioxo-1,2,3,5,6,8,10,13a-octahydrocyclopenta[b][1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-11-carboxamide (2.2.3): LC-MS (ESI) 46e (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.28 (s, 1H), 10.42 (t, J=5.8 Hz, 1H), 8.38 (s, 1H), 7.34 (d, J=8.4 Hz, 1H), 4.54 (m, 2H), 4.34 (m, 3H), 3.75 (dd, J.sub.1=8.0 Hz, J.sub.2=6.0 Hz, 1H), 2.30 (m, 1H), 2.12 (m, 1H), 1.76 (m, 3H), 1.36 (d, J=6.0 Hz, 3H).
Example 3. General Synthetic Procedure for Annelated 9-hydroxy-1,8-dioxo-1,3,4,8-tetrahydro-2H-pyrido[1,2-a]pyrazine-7-carboxamides of General Formula 1.1
[0237] (3S,11aR)-6-hydroxy-N-[(2,6-difluoropyridin-3-yl)methyl]-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.19) is prepared according to Scheme 3.
##STR00024##
[0238] ((3S,11aR)-6-(benzyloxy)-8-bromo-3-methyl-2,3,11,11a-tetrahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-5,7-dione (3.1) (3 g, 7.4 mmol) was dissolved in a mixture of THF (250 ml) and methanol (150 ml). The reaction mixture was transferred to an autoclave and triethylamine (1.3 ml, 8.9 mmol) and Pd(dppf)Cl.sub.2 (0.05 g, 0.07 mmol) were added. CO was pumped to 15 atm, and the reaction mixture was heated to 100° C. for 3 days. Upon completion of the reaction (LC-MS control), the reaction mass was filtered through celite, evaporated in vacuum, dissolved in dichloromethane, washed with water, dried over sodium sulfate, and evaporated on a rotary evaporator. The residue was subjected to column chromatography on silica gel with dichloromethane-methanol (60:1) used as the eluent to yield 1.8 g (63%) of methyl (3S,11aR)-6-(benzyloxy)-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]-oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxylate (3.2): .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 8.39 (s, 1H), 7.53 (m, 2H), 7.34 (m, 3H), 5.32 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 5.04, 5.17 (ABq, J.sub.AB=10.4 Hz, 2H), 4.67 (dd, J.sub.1=12.0 Hz, J.sub.2=3.2 Hz, 1H), 4.29 (m, 2H), 3.94 (dd, J.sub.1=12.0 Hz, J.sub.2=10.0 Hz, 1H), 3.63 (dd, J.sub.1=8.0 Hz, J.sub.2=6.4 Hz, 1H), 1.27 (d, J=6.0 Hz).
[0239] A solution of LiOH x H.sub.2O (0.33 g, 7.8 mmol) in 5 ml of water was added to a solution of methyl ether 3.2 (1.5 g, 3.9 mmol) in 15 ml of dioxane. The mixture was then stirred for 3 h at 50° C. Dioxane was evaporated on a rotary evaporator. The residue was diluted with water and acidified with 10% sulfuric acid to pH 3. The precipitate was filtered off, washed with water, and dried in vacuum to yield 1 g (69%) of (3S,11aR)-6-(benzyloxy)-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxylic acid (3.3): .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 15.45 (brs, 1H), 8.75 (s, 1H), 7.54 (m, 2H), 7.36 (m, 3H), 5.38 (dd, J.sub.1=10.0 Hz, J.sub.2=3.2 Hz, 1H), 5.15, 5.25 (ABq, J.sub.AB=10.4 Hz, 2H), 4.87 (dd, J.sub.1=12.0 Hz, J.sub.2=3.2 Hz, 1H), 4.32 (m, 2H), 4.13 (t, J=11.0 Hz, 1H), 3.66 (dd, J.sub.1=7.6 Hz, J.sub.2=6.4 Hz, 1H), 1.28 (d, J=6.0 Hz).
[0240] Triethylamine (1.2 ml, 8.1 mmol) was added to a solution of acid 3.3 (1 g, 2.7 mmol) in 20 ml of dichloromethane. The reaction mixture was stirred at room temperature for 15 minutes and TBTU (0.82 g, 3.3 mmol) was added. The mixture was stirred for another 15 min, then [(2,6-difluoropyridin-3-yl)methyl]amine (0.47 g, 3.3 mmol) was added, and the mixture was stirred for 14 h at room temperature. The reaction mass was washed with 10-% aqueous solution of potassium carbonate and water. The organic layer was dried over sodium sulfate and evaporated on a rotary evaporator. Diethyl ether was added to the residue, and the resulting precipitate was filtered off and dried in vacuum. The product was further used without extra purification. The yield was 0.94 g (70%) of (3S,11aR)-6-(benzyloxy)-3-methyl-5,7-dioxo-N-[(2,6-difluoropyridin-3-yl)methyl]-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (3.4.19): .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 10.45 (t, J=5.9 Hz, 1H), 8.56 (s, 1H), 8.13-8.00 (m, 1H), 7.57-7.52 (m, 2H), 7.40-7.29 (m, 3H), 7.15 (dd, J.sub.1=8.1 Hz, J.sub.2=2.3 Hz, 1H), 5.35 (dd, J.sub.1=10.0 Hz, J.sub.2=3.7 Hz, 1H), 5.21 (d, J=10.6 Hz, 1H), 5.07 (d, J=10.6 Hz, 1H), 4.79 (dd, J.sub.1=12.2 Hz, J.sub.2=3.7 Hz, 1H), 4.56 (d, J=5.9 Hz, 2H), 4.38-4.22 (m, 2H), 4.08-3.98 (m, 1H), 3.64 (dd, J.sub.1=8.1 Hz, J.sub.2=6.4 Hz, 1H), 1.27 (d, J=6.1 Hz, 3H).
[0241] Compound 3.4.19 (0.9 g, 1.8 mmol) was dissolved in a mixture of THF (18 ml) and methanol (2 ml), then 10% Pd/C (0.09 g) was added, and the reaction mixture was stirred in a hydrogen atmosphere for 8 h. The reaction mass was passed through celite, the filtrate and then evaporated. The residue was treated with ether, the precipitate was filtered off and dried in vacuum to yield 0.6 g (81%) of (3S,11aR)-6-hydroxy-N-[(2,6-difluoro-pyridin-3-ylmethyl]-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.19): LC-MS (ESI) 07 (M+H).sup.+; .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.49 (brs, 1H), 10.36 (t, J=5.8 Hz, 1H), 8.45 (s, 1H), 8.06 (dd, J.sub.1=17.2 Hz, J.sub.2=8.4 Hz, 1H), 7.14 (dd, J.sub.1=8.0 Hz, J.sub.2=2.0 Hz, 1H), 5.38 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 4.88 (dd, J.sub.1=12.0 Hz, J.sub.2=3.6 Hz, 1H), 4.55 (d, J=5.6 Hz, 2H), 4.39 (dd, J.sub.1=8.0 Hz, J.sub.2=6.8 Hz, 1H), 4.29 (m, 1H), 4.00 (t, J=11.0 Hz, 1H), 3.67 (dd, J.sub.1=8.5 Hz, J.sub.2=6.8 Hz, 1H), 1.34 (d, J=6.4 Hz, 3H); XRD: Simulation of the diffraction pattern of an inhibitor 1.1.19 sample using the Pauli technique is shown in
[0242] Sodium (3S, 11aR)-8-({[(2,6-difluoropyridin-3-yl)methyl]amino}carbonyl)-3-methyl-5,7-dioxo-2,3,5,7,11, 11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazin-6-olate (1.1.20). NaOH (0.05 g, 1.2 mmol) is added to a suspension of (3S,11aR)-6-hydroxy-N-[(2,4-difluoro-3-pyridyl)methyl]-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazole[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.19) (0.05 g, 1.2 mmol) in 80 ml of ethanol and 20 ml of water at 75° C. The reaction mixture is stirred for 14 h at room temperature. The resulting precipitate is filtered off, washed with absolute ethanol, and dried in vacuum to yield 0.42 g (80%) of sodium (3S, 11aR)-8-({[(2,6-difluoropyridin-3-yl)methyl]amino}carbonyl)-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazin-6-olate (1.1.20): .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 10.76 (t, J=5.6 Hz, 1H), 8.00 (dd, J.sub.1=17.2 Hz, J.sub.2=8.0 Hz, 1H), 7.90 (s, 1H), 7.11 (dd, J.sub.1=8.0 Hz, J.sub.2=1.6 Hz, 1H), 5.21 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.59 (dd, J.sub.1=12.4 Hz, J.sub.2=3.6 Hz, 1H), 4.52 (t, J=5.6 Hz, 2H), 4.25 (m, 2H), 3.74 (t, J=10.8 Hz, 1H), 3.60 (m, 1H), 1.27 (d, J=5.2 Hz, 3H).
[0243] The following compounds are obtained in a similar way:
[0244] (3S,11aR)-6-hydroxy-3-methyl-5,7-dioxo-N-(2-thienylmethyl-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.1). LC-MS (ESI) 376 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.47 (brs, 1H), 10.33 (t, J=4.8 Hz, 1H), 8.49 (s, 1H), 7.40 (dd, J.sub.1=8.8 Hz, J.sub.2=4.8 Hz, 1H), 7.03 (d, J=2.8 Hz, 1H), 6.96 (dd, J.sub.1=4.8 Hz, J.sub.2=3.6 Hz, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.90 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.70 (d, J=5.6 Hz, 2H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 4.29 (m, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.67 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 1.34 (d, J=6.0 Hz, 3H);
[0245] (3S,11aR)-6-hydroxy-3-methyl-N-[(4-methyl-4H-1,3-oxazol-2-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.14). LC-MS (ESI) 375 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 300 MHz, 80° C.) δ 10.49 (brs, 1H), 7.93 (s, 1H), 5.21 (dd, J.sub.1=9.6 Hz, J.sub.2=3.6 Hz, 1H), 4.58-4.85 (m, 3H), 4.29 (m, 2H), 3.64 (m, 2H), 2.30 (s, 3H), 1.30 (d, J=5.4 Hz, 3H);
[0246] (3S,11aR)-6-hydroxy-3-methyl-N-[(1,4,5-trimethyl-1H-imidazol-2-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.10): LC-MS (ESI) 402 (M+H).sup.+;
[0247] (3S,11aR)-6-hydroxy-3-methyl-N-[(4-methyl-1,2,3-thiadiazol-5-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.11): LC-MS (ESI) 392 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.52 (brs, 1H), 10.45 (t, J=5.6 Hz, 1H), 8.49 (s, 1H), 5.39 (dd, J.sub.1=9.2 Hz, J.sub.2=3.2 Hz, 1H), 4.89 (m, 1H), 4.81 (d, J=5.6 Hz, 2H), 4.40 (dd, J.sub.1=7.6 Hz, J.sub.2=0.4 Hz, 1H), 4.29 (m, 1H), 4.00 (t, J=11.2 Hz, 1H), 3.66 (t, J=7.6 Hz, 1H), 2.67 (s, 3H), 1.34 (d, J=6.0 Hz, 3H);
[0248] (3S,11aR)-6-hydroxy-3-methyl-N-[(3-methyl-1,2,4-oxadiazol-5-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.13): LC-MS (ESI) 376 (M+H).sup.+;
[0249] (3S,11aR)-6-hydroxy-3-methyl-N-[(3-fluoropyridin-2-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.16): LC-MS (ESI) 389 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 10.61 (t, J=5.4 Hz, 1H), 8.57 (s, 1H), 8.42 (d, J=4.5 Hz, 1H), 7.77-7.67 (m, 1H), 7.57-7.50 (m, 2H), 7.47-7.28 (m, 4H), 5.40-5.31 (m, 1H), 5.25-5.17 (m, 1H), 5.11-5.03 (m, 1H), 4.84-4.70 (m, 3H), 4.37-4.23 (m, 2H), 4.08-3.97 (m, 1H), 3.68-3.60 (m, 1H), 1.28 (d, J=6.1 Hz, 3H);
[0250] (3S,11aR)-6-hydroxy-N-[(3,5-difluoropyridin-2-yl)methyl]-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.17): LC-MS (ESI) 407 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.42 (brs, 1H), 10.54 (t, J=5.2 Hz, 1H), 8.49 (d, J=2.4 Hz, 1H), 8.46 (s, 1H), 7.95 (dt, J.sub.1=11.4 Hz, J.sub.2=2.4 Hz, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.88 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.71 (d, J=4.8 Hz, 2H), 4.39 (dd, J.sub.1=8.0 Hz, J.sub.2=6.8 Hz, 1H), 4.29 (m, 1H), 4.00 (t, J=11.2 Hz, 1H), 3.66 (dd, J.sub.1=8.0 Hz, J.sub.2=6.8 Hz, 1H), 1.34 (d, J=6.4 Hz, 3H);
[0251] (3S,11aR)-6-hydroxy-N-[(6-chloropyridin-3-yl)methyl]-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.18): LC-MS (M+1)=405, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 10.76 (t, J=5.8 Hz, 1H), 8.35 (s, 1H), 7.92 (s, 1H), 7.75 (d, J=8.0 Hz, 1H), 7.45 (d, J=8.0 Hz, 1H), 5.21 (m, 1H), 4.60 (m, 1H), 4.53 (m, 2H), 4.25 (m, 2H), 3.75 (t, J=10.8 Hz, 1H), 3.59 (m, 1H), 1.26 (d, J=4.4 Hz, 3H);
[0252] Sodium (3S, 11aR)-8-({[(2,6-difluoropyridin-3-yl)methyl]amino}carbonyl)-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazin-6-olate 1.1.20): LC-MS (ESI) 407 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.49 (brs, 1H), 10.36 (t, J=5.8 Hz, 1H), 8.45 (s, 1H), 8.06 (dd, J.sub.1=17.2 Hz, J.sub.2=8.4 Hz, 1H), 7.14 (dd, J.sub.1=8.0 Hz, J.sub.2=2.0 Hz, 1H), 5.38 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 4.88 (dd, J.sub.1=12.0 Hz, J.sub.2=3.6 Hz, 1H), 4.55 (d, J=5.6 Hz, 2H), 4.39 (dd, J.sub.1=8.0 Hz, J.sub.2=6.8 Hz, 1H), 4.29 (m, 1H), 4.00 (t, J=11.0 Hz, 1H), 3.67 (dd, J.sub.1=8.5 Hz, J.sub.2=6.8 Hz, 1H), 1.34 (d, J=6.4 Hz, 3H);
[0253] (3S,11aR)-6-hydroxy-3-methyl-N-[(2,4,6-trifluoropyridin-3-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.22): LC-MS (ESI) 425 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.46 (s, 1H), 10.36 (t, J=6.0 Hz, 1H), 8.43 (s, 1H), 7.33 (d, J=8.4 Hz, 1H), 5.37 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.85 (dd, J.sub.1=12.4 Hz, J.sub.2=4.0 Hz, 1H), 4.55 (d, J=6.0 Hz, 2H), 4.39 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 4.28 (sxt, J=6.4 Hz, 1H), 3.99 (dd, J.sub.1=11.6 Hz, J.sub.2=10.8 Hz, 1H), 3.66 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 1.33 (d, J=6.4 Hz, 3H);
[0254] (3S,11aR)-6-hydroxy-3-methyl-N-[(3-fluoropyridin-4-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.23): LC-MS (ESI) 389 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 10.54-10.44 (m, 1H), 8.57 (s, 1H), 8.52 (s, 1H), 8.38 (d, J=4.4 Hz, 1H), 7.55 (d, J=7.2 Hz, 2H), 7.42-7.27 (m, 4H), 5.40-5.30 (m, 1H), 5.23 (d, J=10.4 Hz, 1H), 5.08 (d, J=10.4 Hz, 1H), 4.84-4.74 (m, 1H), 4.65 (d, J=5.6 Hz, 2H), 4.38-4.22 (m, 2H), 4.10-3.96 (m, 1H), 3.70-3.59 (m, 1H), 1.28 (d, J=5.7 Hz, 3H);
[0255] (3S,11aR)-6-hydroxy-N-[(3,5-difluoropyridin-4-yl)methyl]-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.24). LC-MS (ESI) 407 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.46 (brs, 1H), 10.43 (t, J=6.0 Hz, 1H), 8.50 (s, 2H), 8.43 (s, 1H), 5.37 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.84 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.67 (d, J=6.0 Hz, 2H), 4.39 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 4.28 (m, 1H), 3.99 (dd, J.sub.1=12.0 Hz, J.sub.2=6.4 Hz, 1H), 3.66 (dd, J.sub.1=8.8 Hz, J.sub.2=6.8 Hz, 1H), 1.33 (d, J=6.0 Hz, 3H);
[0256] (3S,11aR)-6-hydroxy-3-methyl-N-(pyrazin-2-ylmethyl)-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.25): LC-MS (ESI) 372 (M+H).sup.+, H NMR (DMSO-d.sub.6, 400 MHz) δ 11.45 (brs, 1H), 10.53 (t, J=5.6 Hz, 1H), 8.63 (s, 1H), 8.61 (d, J=2.0 Hz, 1H), 8.54 (d, J=2.0 Hz, 1H), 8.47 (s, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.89 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.71 (d, J=5.6 Hz, 2H), 4.40 (t, J=7.6 Hz, 1H), 4.30 (m, 1H), 4.01 (t, J=11.0 Hz, 1H), 3.67 (dd, J.sub.1=8.0 Hz, J.sub.2=7.2 Hz, 1H), 1.34 (d, J=6.0 Hz, 3H);
[0257] (3S,11aR)-6-hydroxy-3-methyl-N-[(2-chloroimidazo[2,1-b][1,3]thiazol-6-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide 1.1.28: LC-MS (ESI) 450 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 10.31 (brs, 1H), 8.06 (s, 1H), 7.57 (s, 1H), 5.25 (brs, 1H), 4.65 (m, 1H), 4.48 (brs, 2H), 4.28 (brs, 2H), 3.77 (brs, 1H), 3.63 (brs, 1H), 1.29 (brs, 3H);
[0258] (3S,11aR)-6-hydroxy-N-(imidazo[2,1-b][1,3,4]thiadiazol-6-ylmethyl-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.29): LC-MS (ESI) 417 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 10.64 (brs, 1H), 9.18 (s, 1H), 8.00 (s, 1H), 7.91 (s, 1H), 5.22 (m, 1H), 4.60 (m, 1H), 4.49 (s, 2H), 4.25 (s, 2H), 3.75 (m, 1H), 3.59 (m, 1H), 1.26 (brs, 3H);
[0259] (3S,11aR)-6-hydroxy-3-methyl-N-[(2-methylimidazo[2,1-b][1,3,4]thiadiazol-6-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide 1.1.30: LC-MS (ESI) 431 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.43 (brs, 1H), 10.27 (t, J=5.2 Hz, 1H), 8.47 (s, 1H), 7.95 (s, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 4.90 (dd, J.sub.1=12.0 Hz, J.sub.2=3.6 Hz, 1H), 4.49 (d, J=5.2 Hz, 2H), 4.39 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 4.29 (m, 1H), 4.01 (t, J=11.2 Hz, 1H), 3.66 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 2.70 (s, 3H), 1.34 (d, J=6.0 Hz, 3H);
[0260] (3S,11aR)-6-hydroxy-3-methyl-N-[(1-methyl-benzimidazol-2-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.42): LC-MS (ESI) 424 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.49 (brs, 1H), 10.60 (t, J=4.8 Hz, 1H), 8.44 (s, 1H), 7.68 (d, J=8.0 Hz, 1H), 7.63 (d, J=7.6 Hz, 1H), 7.37 (t, J=8.0 Hz, 1H), 7.63 (t, J=7.6 Hz, 1H), 5.39 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 4.95 (d, J=4.8 Hz, 2H), 4.88 (m, 1H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 4.30 (m, 1H), 3.99 (t, J=11.2 Hz, 1H), 3.89 (s, 3H), 3.67 (dd, J.sub.1=8.4 Hz, J.sub.2=6.8 Hz, 1H), 1.35 (d, J=6.4 Hz, 3H);
[0261] (3S,11aR)-6-hydroxy-3-methyl-N-[(6-chloro-1-isopropyl-1H-benzimidazol-2-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide 1.1.45): LC-MS (ESI) 486 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.47 (brs, 1H), 10.57 (t, J=5.2 Hz, 1H), 8.51 (s, 1H), 7.80 (d, J=2.0 Hz, 1H), 7.61 (d, J=8.8 Hz, 1H), 7.20 (dd, J.sub.1=8.8 Hz, J.sub.2=2.0 Hz, 1H), 5.40 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 4.91 (dd, J.sub.1=12.0 Hz, J.sub.2=4.0 Hz, 1H), 4.86 (d, J=5.2 Hz, 2H), 4.83 (m, 1H), 4.40 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 4.29 (m, 1H), 4.02 (dd, J.sub.1=12.0 Hz, J.sub.2=10.0 Hz, 1H), 3.66 (dd, J.sub.1=8.4 Hz, J.sub.2=7.2 Hz, 1H), 1.54 (d, J=6.8 Hz, 6H), 1.34 (d, J=6.0 Hz, 3H);
[0262] (3S,11aR)-N-(2,1,3-benzothiadiazol-5-ylmethyl)-6-hydroxy-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.47). LC-MS (ESI) 428 (M+H).sup.+;
[0263] (3S,11aR)-6-hydroxy-3-methyl-N-(quinolin-6-yl)methyl)-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.53): LC-MS (ESI) 421 (M+H).sup.+;
[0264] (3S,11aR)-6-hydroxy-N-(soquinolin-1-ylmethyl)-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.55): LC-MS (ESI) 421 (M+H).sup.+, H NMR (DMSO-d.sub.6, 300 MHz, 80° C.) δ 11.25 (brs, 1H), 10.64 (t, J=5.2 Hz, 1H), 8.47 (m, 2H), 8.32 (d, J=8.0 Hz, 1H), 7.99 (d, J=8.0 Hz, 1H), 7.75 (m, 3H), 5.42 (dd, J.sub.1=10.0 Hz, J.sub.2=3.6 Hz, 1H), 5.20 (d, J=5.2 Hz, 2H), 4.85 (dd, J.sub.1=12.0 Hz, J.sub.2=3.6 Hz, 1H), 4.38 (m, 2H), 4.01 (t, J=10.8 Hz, 1H), 3.70 (m, 1H), 1.38 (d, J=5.6 Hz, 3H);
[0265] (3S,11aR)-6-hydroxy-N-(imidazo[1,2-a]pyridin-3-yl)methyl-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.56): LC-MS (M+1)=410, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 10.67 (t, J=5.6 Hz, 1H), 8.48 (d, J=6.4 Hz, 1H), 7.96 (s, 1H), 7.57 (m, 2H), 7.23 (t, J=8.0 Hz, 1H), 6.92 (t, J=7.2 Hz, 1H), 5.20 (dd, J.sub.1=9.6 Hz, J.sub.2=3.6 Hz, 1H), 4.89 (m, 2H), 4.61 (m, 1H), 4.23 (m, 2H), 3.75 (t, J=10.8 Hz, 1H), 3.58 (m, 1H), 1.24 (d, J=5.6 Hz, 3H);
[0266] (3S,11aR)-6-hydroxy-3-methyl-N-(1,2,4-triazolo[4,3-a]pyridin-3-ylmethyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.57): LC-MS (ESI) 411 (M+H).sup.+;
[0267] (3S,11aR)-6-hydroxy-N-(imidazo[1,2-a]pyrimidin-2-ylmethyl)-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.58): LC-MS (ESI) 411 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.46 (brs, 1H), 10.41 (t, J=5.6 Hz, 1H), 8.94 (d, J=5.6 Hz, 1H), 8.50 (m, 2H), 7.79 (s, 1H), 7.03 (m, 1H), 5.40 (m, 1H), 4.91 (m, 1H), 4.67 (d, J=5.6 Hz, 2H), 4.39 (dd, J.sub.1=8.0 Hz, J.sub.2=7.2 Hz, 1H), 4.30 (m, 1H), 4.02 (m, 1H), 3.67 (dd, J.sub.1=8.0 Hz, J.sub.2=7.2 Hz, 1H), 1.34 (d, J=6.0 Hz, 3H);
[0268] (3S,11aR)-6-hydroxy-N-(imidazo[1,2-a]pyrazin-3-ylmethyl)-3-methyl-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide (1.1.59): LC-MS (ESI) 411 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 300 MHz, 80° C.) δ 10.39 (brs, 1H), 8.98 (s, 1H), 8.59 (d, J=6.0 Hz, 1H), 8.07 (brs, 1H), 7.88 (d, J=6.0 Hz, 1H), 7.76 (s, 1H), 5.23 (m, 1H), 4.89 (m, 2H), 4.65 (m, 1H), 4.26 (s, 2H), 3.72 (brs, 1H), 3.60 (m, 1H), 1.28 (d, J=6.0 Hz, 3H);
[0269] (3S,11aR)-6-hydroxy-3-methyl-N-[(6-chloro-1,2,4-triazolo[4,3-b]pyridazin-3-yl)methyl]-5,7-dioxo-2,3,5,7,11,11a-hexahydro[1,3]oxazolo[3,2-a]pyrido[1,2-d]pyrazine-8-carboxamide 1.1.60): LC-MS (ESI) 446 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 11.50 (brs, 1H), 10.59 (brs, 1H), 8.47 (m, 2H), 7.52 (d, J=9.6 Hz, 1H), 5.38 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 5.04 (d, J=5.6 Hz, 2H), 4.88 (m, 1H), 4.39 (dd, J.sub.1=8.2 Hz, J.sub.2=7.2 Hz, 1H), 4.29 (sxt, J=6.4 Hz, 1H), 4.00 (t, J=11.0 Hz, 1H), 3.66 (dd, J.sub.1=8.2 Hz, J.sub.2=7.2 Hz, 1H), 1.34 (d, J=6.4 Hz, 3H);
Example 4. General Synthetic Procedure for Annelated 9-hydroxy-1,8-dioxo-1,3,4,8-tetrahydro-2H-pyrido[1,2-a]pyrazine-7-carboxamides of General Formula 1.2 or 1.3
[0270] Inhibitors 1.2 and 1.3 are prepared similarly to the synthesis of compounds of general formula 1.1 (Example 3) starting from (4R,12aS)-7-benzyloxy-9-bromo-4-methyl-6,8-dioxo-3,4,6,8,12,12a-hexahydro-2H-pyrido[1′,2′:4,5]pyrazino-[2,1-b][1,3]oxazine and (2R,5S,13aR)-8-benzyloxy-10-bromo-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopurido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine, respectively, including as follows:
[0271] (4R,12aS)-7-hydroxy-N-[(2,6-difluoropyridin-3-yl)methyl]-4-methyl-6,8-dioxo-3,4,6,8,12,12a-hexahydro-2H-pyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazine-9-carboxamide 1.2.1: LC-MS (ESI) 421 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.51 (brs, 1H), 10.40 (t, J=5.8 Hz, 1H), 8.48 (s, 1H), 8.05 (q, J=8.8 Hz, 1H), 7.14 (dd, J.sub.1=8.0 Hz, J.sub.2=2.4 Hz, 1H), 5.44 (m, 1H), 4.79 (m, 1H), 4.55 (m, 3H), 4.34 (dd, J.sub.1=13.6 Hz, J.sub.2=5.6 Hz, 1H), 4.03 (dt, J.sub.1=12.0 Hz, J.sub.2=1.6 Hz, 1H), 3.89 (m, 1H), 2.01 (m, 1H), 1.55 (m, 1H), 1.33 (d, J=7.2 Hz, 3H);
[0272] (4R,12aS)-7-hydroxy-4-methyl-N-[(2,4,6-trifluoropyridin-3-yl)methyl]-6,8-dioxo-3,4,6,8,12,12a-hexahydro-2H-pyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazine-9-carboxamide (1.2.3): LC-MS (ESI) 439 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.49 (brs, 1H), 10.39 (t, J=6.0 Hz, 1H), 8.46 (s, 1H), 7.33 (d, J=8.4 Hz, 1H), 5.43 (dd, J.sub.1=9.6 Hz, J.sub.2=4.0 Hz, 1H), 4.78 (p, J=6.6 Hz, 1H), 4.53 (m, 3H), 4.32 (dd, J.sub.1=13.6 Hz, J.sub.2=5.6 Hz, 1H), 4.02 (t, J=12.0 Hz 1H), 3.88 (m, 1H), 2.01 (m, 1H), 1.54 (m, 1H), 1.32 (d, J=7.2 Hz, 3H);
[0273] (4R,12aS)-7-hydroxy-N-[(3,5-difluoropyridin-4-yl)methyl]-4-methyl-6,8-dioxo-3,4,6,8,12,12a-hexahydro-2H-pyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazine-9-carboxamide (1.2.4): LC-MS (ESI) 421 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.49 (s, 1H), 10.47 (t, J=5.6 Hz, 1H), 8.50 (s, 2H), 8.45 (brs, 1H), 5.43 (m, 1H), 4.78 (m, 1H), 4.66 (d, J=5.6 Hz, 2H), 4.53 (m, 1H), 4.32 (m, 1H), 4.02 (t, J.sub.1=12.0 Hz, 1H), 3.89 (m, 1H), 2.00 (m, 1H), 1.54 (d, J=13.6 Hz, 1H), 1.32 (d, J=7.2 Hz, 3H);
[0274] (2R,5S,13aR)-8-hydroxy-N-[(2,6-difluoropyridin-3-yl)methyl]-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopurido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide (1.3.1): LC-MS (M+1)=433, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.47 (s, 1H), 10.41 (m, 1H), 8.45 (s, 1H), 8.04 (m, 1H), 7.15 (d, J=7.2 Hz, 1H), 5.44 (m, 1H), 5.10 (brs, 1H), 4.68 (m, 1H), 4.60 (brs, 1H), 4.55 (m, 2H), 4.03 (m, 1H), 1.94 (brs, 4H), 1.84 (m, 1H), 1.58 (m, 1H);
[0275] (2R,5S,13aR)-8-hydroxy-N-[(2,4,6-trifluoropyridin-3-yl)methyl]-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide (1.3.2): LC-MS (ESI) 451 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.46 (s, 1H), 10.40 (t, J=5.6 Hz, 1H), 8.42 (s, 1H), 7.33 (d, J=8.4 Hz, 1H), 5.43 (dd, J.sub.1=10.0 Hz, J.sub.2=4.0 Hz, 1H), 5.09 (brs, 1H), 4.65 (dd, J.sub.1=12.8 Hz, J.sub.2=4.0 Hz, 1H), 4.60 (brs, 1H), 4.54 (m, 2H), 4.01 (dd, J.sub.1=12.4 Hz, J.sub.2=9.6 Hz, 1H), 1.94 (brs, 4H), 1.84 (d, J=12.0 Hz, 1H), 1.57 (m, 1H);
[0276] (2R,5S,13aR)-8-hydroxy-N-[(3,5-difluoropyridin-4-yl)methyl]-7,9-dioxo-2,3,4,5,7,9,13,13a-octahydro-2,5-methanopyrido[1′,2′:4,5]pyrazino[2,1-b][1,3]oxazepine-10-carboxamide (1.3.3): LC-MS (ESI) 433 (M+H).sup.+, .sup.1H NMR (DMSO-d.sub.6, 400 MHz) δ 12.46 (s, 1H), 10.47 (t, J=5.8 Hz, 1H), 8.49 (s, 2H), 8.42 (s, 1H), 5.43 (dd, J.sub.1=9.2 Hz, J.sub.2=4.0 Hz, 1H), 5.09 (brs, 1H), 4.66 (m, 3H), 4.60 (brs, 1H), 4.01 (dd, J.sub.1=12.4 Hz, J.sub.2=9.6 Hz, 1H), 1.94 (brs, 4H), 1.84 (d, J=12.0 Hz, 1H), 1.57 (m, 1H);
Example 5. A Pharmaceutical Composition in the Form of a Tablet
[0277] Starch (1600 mg), ground lactose (1600 mg), talc (400 mg), and 900 mg of the compound of general formula 1 or 2 were mixed and pressed into a bar. The resulting bar was crushed into granules and sieved through a sieve to collect granules of 14-16 mesh size. The granules thus obtained were formed into tablets of a suitable form weighing 225 or 450 mg each.
Example 6. A Pharmaceutical Composition in the Form of Capsules
[0278] Compounds of general formula 1 or 2 were thoroughly mixed with lactose powder in a 2:1 ratio. The resulting powdery mixture was packaged in suitable gelatin capsules of 30, 60, or 120 mg in each capsule.
Example 7. A Lyophilized Pharmaceutical Nanocomposition
[0279] Zirconium sand (150 ml) (grinding media: 0.5 mm YTZ® Zirconia Grinding and Dispersion Media; Tosoh USA, Inc.) and a solution of poloxamer P338 (10.0 g) and sucrose or mannitol (11.6 g) in 400 ml of phosphate buffered saline (PBS, pH=7.4) were loaded into a container (Jar). The resulting solution (150 ml) was placed in a jar and zirconium sand (150 ml) (grinding media: 0.5 mm YTZ® Zirconia Grinding and Dispersion Media; Tosoh USA, Inc.) and a compound of general formula 1 or 2 (˜15 g) were added. The jar was placed on a jar mill (U.S. Stoneware 700 Series), the rotation speed was set at 104 rpm, and the mixture was ground for 24 hours. The end of the process was controlled by particle size distribution measurements on a Malvern Zetasizer Nano ZS instrument. At the end of the grinding process, the rotation was stopped and the mixture was left for 16-20 hours at 2-8° C. to let the zirconium sand precipitate. The suspension was filtered through a glass filter with a porosity of 5-15 microns and analyzed to determine the content (concentration) of the compound of general formula 1 or 2. The resulting nanosuspension with a particle size of 200 to 900 nm, preferably 200-250 nm, was poured in sterile conditions at 2 ml in 5-ml sterile glass vials, frozen and lyophilized to yield a pharmaceutical nanocomposition in the form of a lyophilizate.
Example 8. Injectable Drug
[0280] A previously prepared sterile PBS solution with pH=6.8 was added to the nanocomposition obtained in Example 7 in the form of a lyophilizate at the rate of 2.2 ml per 5 ml vial. The vials were stoppered with pre-sterilized plugs and then sealed. The resulting pharmaceutical nanosuspension is ready for use.
Example 9. Injectable Drug
[0281] A compound of general formula 1 or 2 was sterilized with gamma radiation and subjected to clean milling of wet granules with mannite, polysorbate 20, polyethylene glycol 3350, and water for injection to achieve an average cabotegravir particle size of 200 nm. The nanosuspension was placed in pre-sterilized glass vials. The vials were stoppered with pre-sterilized plugs and then sealed. The resulting pharmaceutical nanosuspension is ready for use.
Example 10. Anti-HIV Activity of Compounds of General Formula 1 or 2 and the Prototype (CAB)
[0282] a) Inhibitory Activity of the Lymphocytropic Virus Strain HIV-1 (IIIB) in the CEM-SS Cell Line.
[0283] The evaluation was done by ImQuest BioSciences, Inc. (Frederick, Md., USA). CEM-SS cells [Foley G. E. et al. Continuous Culture of Human Lymphoblasts from Peripheral Blood of a Child with Acute Leukemia. Cancer 1965, 18, 522-529] obtained through the NIH AIDS Research and Reference Reagent Program (Manassas, Va., USA) were passaged in T-75 flasks prior to use cell culture medium consisting of RPMI1640 medium (Lonza; Walkersville, Md., USA) supplemented with 10% heat inactivated fetal bovine serum (Gibco; Grand Island, N.Y.), 2 μM L-glutamine (Lonza), 100 U/μl penicillin (Lonza) and 100 μg/μl streptomycin (Lonza) for antiviral analysis. The CEM-SS cells were kept in RPMI 140 medium supplemented with 10% heat inactivated bovine fetal serum, 2 mM glutamine, 100 U/ml penicillin, and 100 μg/ml streptomycin.
[0284] A lymphocytropic HIV-1 IIIB strain [Popovic, M. et al. Detection, isolation, and continuous production of cytopathic retroviruses (HTLV-III) from patients with AIDS and pre-AIDS. Science. 1984, 224 (4648), 497-500] was chosen for a virus to be analyzed. The virus was obtained through the NIH AIDS Research and Reference Reagents Program and grown in CEM-SS cells to produce a stock virus pool. A pre-titrated aliquot of the virus was removed from the freezer (−80° C.) and allowed to slowly thaw to room temperature in a biological safety cabinet. The virus was diluted in a tissue culture medium so that the amount of virus added in 50 μl per well was sufficient to kill 85 to 95% of the cells 6 days after infection. HIV-1 IIIB was grown and titrated in CEM-SS cells. The inhibitory effect of CEM-SS compounds on HIV-1 compounds was evaluated in CEM-SS cells as described earlier [Nara, P. L. at al. Simple, rapid, quantitative, syncytium-forming microassay for the detection of human immunodeficiency virus neutralizing antibody. AIDS Res. Hum. Retroviruses 1987, 3, 283-302.] in microtiter plate anti-HIV assays to quantitatively assess the ability of a compound to inhibit HIV-induced cell death. Quantification was carried out using 2,3-bis-(2-methoxy-4-nitro-5-sulfenyl)-(2H)-tetrazolium-5-carboxanilide (XTT) tetrazolium, which is metabolized to a stained formazan product through viable cell.sub.SI-O The compounds given in Table 1 were dissolved at 10 μM in DMSO and stored at −20° C.
[0285] Test materials were evaluated using antiviral assays in triplicate in concentrations of 30 and 10 nM. The compounds shown in table 1 were dissolved at 10 μM in DMSO and stored at −20° C. Test materials were evaluated using concentrations of 30 and 10 nM in triplicate for antiviral assays. Sixty nanomolar concentrations of each compound (2 times that of the test concentration in the well) were prepared by diluting 10 μM DMSO stock 1:100 in cell culture medium (10 μl of compound added to 990 μl of the medium) followed by 1:10 dilution (10 μl of the compound was added to 90 μl of the medium), then 6 μl was added to 994 μl of the medium; 100 100 μl was added to a 96-well microtiter plate in triplicate to ensure efficacy and one colorimetric well. Twenty nanomolar concentrations of each compound (twice as much as the test concentration in the well) were prepared by diluting 10 μM DMSO with the original 1:100 in cell culture medium (10 μl of the compound added to 990 μl of the medium) followed by diluting 1:100 (10 μl of the compound was added to 900 μl of the medium), then 20 μl was added to 980 μl of the medium; 100 μl was added to a 96-well microtiter plate in triplicate to ensure efficacy and one colorimetric well.
[0286] Azidothymidine (AZT) was purchased from Sigma Aldrich (USA) and evaluated as a positive control compound in an antiviral assay using a six-concentration reaction curve on each assay plate.
[0287] Test compounds were initially evaluated for HIV-1 IIIB strain in CEM-SS cells at test concentrations of 30 and 10 nM. Antiviral efficacy data are summarized in Table 1. Compounds that were active at 10 and 30 nM were evaluated in an HIV cytoprotection assay at 2 nM. For compounds showing activity at 2 nM, inhibitory activity (EC.sub.50) was determined. Data on antiviral efficacy are also shown in Table 1.
[0288] The AZT control compound was evaluated in parallel with the test compounds presented and gave EC.sub.50 values ranging from less than 2 nM to 7 nM. When evaluating the antiviral activity of compounds at 2 nM, AZT demonstrated EC.sub.50 values ranging from 10 to 20 nM.
TABLE-US-00002 TABLE 1 Inhibitory activity (EC.sub.50) against HIV-1 (IIIB strain) demonstrated by compounds of general formula 1 and 2 in the CEM-SS cell line. Compound No EC.sub.50 (nM) 1.1.1 1.44 1.1.3 >2-<10 1.1.4 ~8 1.1.5 >10-<30 1.1.6 >10-<30 1.1.7 0.59 1.1.8 >2-<10 1.1.9 >30 1.1.11 >30 1.1.12 >10-<30 1.1.14 ~10 1.1.15 >10-<30 1.1.16 .sup. >10-<30, 10.5.sup.a 1.1.17 0.24, 1.5.sup.a 1.1.18 >10-<30 1.1.19 0.24, 1.9.sup.a 1.1.20 4.8 1.1.21 >30 1.1.23 >10-<30 1.1.24 2.63 1.1.26 >10-<30 1.1.27 >30 1.1.28 >30 1.1.29 >30 1.1.30 >30 1.1.31 >2-<10 1.1.32 >2-<10 1.1.33 >10-<30 1.1.34 >2-<10 1.1.35 <2 1.1.36 >10-<30 1.1.37 1.93 1.1.38 >30 1.1.39 >2-<10 1.1.40 >30 1.1.41 >10-<30 1.1.42 >30 1.1.43 >30 1.1.44 >30 1.1.45 >30 1.1.46 >30 1.1.48 0.43 1.1.49 >2-<10 1.1.50 >2-<10 1.1.51 >2-<10 1.1.52 ~10 1.1.54 2.17 1.1.55 .sup. >2-<10, 14.5.sup.a 1.1.56 >30 1.1.58 >30 1.1.59 >30 1.1.60 >10-<30 1.1.61 >30 1.1.62 >2-<10 1.1.63 >30 1.1.64 >30 1.1.65 <2 1.2.1 5.15 1.2.3 >2-<10 1.2.4 >2-<10 1.3.1 >2-<10 1.3.2 >2-<10 1.3.3 >2-<10 2.1.1 >2-<10 2.1.2 >2-<10 2.1.3 >2-<10 2.2.1 ~10 2.2.3 >2-<10 .sup.aInhibitory activity of HIV-1 (NL4.3 strain) carrying the GFP reporter gene (NL4.3-GFP) in the SupT1 cell line.
[0289] 6. Inhibitory Activity of HIV-1 (NL4.3 Strain) Carrying the GFP Reporter Gene (NL4.3-GFP) in the SuPT1 Cell Line.
[0290] The evaluation was done by RetroVirox Inc. (San Diego, Calif., USA). SupT1 cells were infected with the HIV NL4.3 strain carrying the green fluorescent protein gene (NL4.3-GFP). The virus sample was produced by transfection of 293T cells with proviral DNA. The sample was frozen 48 hours after transfection and stored until use. To enhance the efficiency of infection, a suspension of SupT1 cells was precipitated from the infectious mixture by centrifugation. Test substances were added to the cells immediately before adding the virus. After 2 hours of incubation, the infectious mixture was replaced with a fresh culture medium containing test samples. The effectiveness of the infection was determined after 45 hours by counting the percentage of fluorescent cells in comparison with uninfected cell cultures. The cytotoxicity of the test compounds was determined in parallel on the same, but not infected, SupT1 cell line using the XTT reagent. Serial tenfold dilutions of the samples (starting from 10 μM when determining antiviral activity or from 100 μM for determining cytotoxicity). As a negative control, 0.1% DMSO was used. The values of EC.sub.50 were calculated. The quality of the tests was determined using the following controls: signal-to-noise ratio, integrase inhibitor raltegravir (1 μM), and test reproducibility. The results are presented in Table 1.
Example 11. Evaluation of Thermodynamic Solubility of Compounds of General Formula 1 or 2
[0291] The test compound of general formula 1 or 2 (2 mg) was weighed into a glass vial. The pION universal buffer (0.5 ml) with pH 7.4 or deionized water was added to appropriate vials. The vials were tightly closed with a lid, placed on a rotating shaker, and incubated for 24 hours to balance the solution and the solid phase at the saturation point. Each resulting solution (200 μl) was taken (in duplicate) from the vials, placed in a 96-well MultiScreen Solubility Filter Plate (Millipore), and filtered using vacuum (10.sup.nHg) into a polypropylene plate with a U-shaped bottom. If necessary, the filtrates were diluted with an appropriate buffer (water). Then, 120 μl of the filtarate (or a diluted filtrate) was transferred to a 96-well plate with a UV-transparent bottom and 60 μl/well of acetonitrile and 20 μl/well of DMSO were added and thoroughly mixed with a multichannel pipette (acetonitrile and DMSO were added to obtain a required composition of calibration standards, as described below). The optical density of the resulting solutions was measured on a multifunction Infinite 200 PRO plate reader in the wavelength range from 240 nm to 400 nm in steps of 10 nm. The value of OD.sub.λ for solutions was assessed at a wavelength chosen for plotting a calibration curve.
[0292] The concentration of a substance in the solution after the solubility test was evaluated using a calibration curve. A stock solution in DMSO with a concentration of 2 mg/ml was prepared from a separate sample. Using the serial dilutions (step 2), six 10-fold solutions in DMSO were prepared, which were then diluted 10 times in an appropriate buffer with 30% acetonitrile to ensure complete solubility of the test compound in the mixture. The range of final concentrations of the calibration curve was 0.2; 0.1; 0.05; 0.025; 0.0125; 0.00625, and 0 mg/ml. Calibration solutions were prepared in a 96-well UV plate as follows: 20 μl of a 10-fold stock solution in DMSO, 60 μl of acetonitrile, and 120 μl of the appropriate buffer were added to the well and stirred. The optical density of the resulting solutions was measured on a multifunction Infinite 200 PRO plate reader in the wavelength range from 240 nm to 400 nm with a step of 10 nm. The optimal wavelength typically corresponding to the absorption maximum was chosen, and a linear dependence of OD at a given wavelength on the concentration of the compound was obtained.
[0293] The concentration of the compound in the filtrate after the solubility test was calculated according to the formula:
shake-flask solubility=(OD.sub.λfiltrate−OD.sub.λblank)/(slope×1.67×filtrate dilution)
[0294] wherein slope means the slope of the calibration curve; 1.67 is the filtrate acetonitrile+DMSO dilution ratio; filtrate dilution is an additional dilution of the filtrate, if applicable. Data on the thermodynamic solubility of compounds of general formulas 1 and 2 are presented in Table 2.
TABLE-US-00003 TABLE 2 Thermodynamic solubility of compounds of general formulas 1 and 2 Wavelength, Water pH 7.4 nm (water/pH Solubility, mg/ml Solubility, mg/ml Compound 7.4) value SD value SD Cabotegravir 290/280 0.008 0.000 0.005 0.000 (CAB) 1.1.1 320/290 0.033 0.000 0.024 0.000 1.1.7 320/280 0.318 0.053 0.318 0.015 1.1.17 320/290 0.131 0.004 0.196 0.011 1.1.19 280/280 0.269 0.003 0.232 0.002 1.1.20 310/280 2.679 0.090 1.684 0.056 1.1.23 320/280 0.078 0.000 0.091 0.002 1.1.24 320/280 0.347 0.011 0.380 0.002 1.1.36 320/290 0.056 0.000 0.070 0.000 1.1.37 310/270 0.021 0.002 0.021 0.001 1.1.48 LC-MSMS 0.00285 0.0000118 0.0032 0.0000118 1.1.51 320/270 0.031 0.001 0.008 0.000 1.1.54 320/300 0.010 0.000 0.006 0.000 1.2.1 320/280 0.057 0.003 0.080 0.000
Example 12. The Pharmacokinetics of Compounds of General Formula 1.1
[0295] The research was carried out on male Sprague-Dawley rats obtained from Charles River Laboratories (Germany).
[0296] The studies were carried out on male Sprague-Dawley rats obtained from Charles River nursery (Germany). The age of the animals at the time of administration was 8-10 weeks. The animals were pre-catheterized so that all blood samples during the study of each substance could be taken from the same animals. Each substance was tested in 3 animals at each route of administration.
[0297] The solution of test substances with a concentration of 0.2 mg/ml in 10% DMSO/10% Solutol/80% 0.05M N-methylglucamine was administered to rats intravenously in the tail vein at a dose of 1 mg/kg. Blood samples were collected at 5, 15, 30 min, 1, 2, 4, 8, and 24 hours after administration. Intragastric administration involved gavage using a solution with a concentration of 0.5 mg/ml in 0.5% methylcellulose at a dose of 5 mg/kg. Blood samples were collected at 15, 30 min, 1, 2, 4, 8, and 24 hours after administration. Blood (0.2 ml) was collected in polypropylene tubes containing 20 μl of 5% NaEDTA. Blood plasma was separated by centrifugation at 1500×g for 10 min and stored until testing at a temperature of −80° C.
[0298] A pure acetonitrile-water (1:1) mixture (5 μl) was added to 45 μl of test samples. The samples were vortexed thoroughly for 10 seconds. Tolbutamide (100 μl) chilled to +4° C. (200 ng/ml in MeCN) was added to the samples and the mixture was vortexed. The samples were precipitated on ice at +4° C. for 15 minutes and then centrifuged at 2700 g for 10 min at +4° C. Supernatants (100 μl each) were transferred to a blank plate for HPLC/MS/MS analysis.
[0299] The HPLC/MS/MS analysis was performed using an Agilent 1290 Infinity system coupled to an AB Sciex QTrap 6500 mass spectrometer.
[0300] Pharmacokinetic analysis of plasma concentration vs time data was performed by a model-independent method using the Phoenix™ WinNonlin® 6.3 software (Pharsight Corp.). The following pharmacokinetic parameters were calculated (Table 3): maximum concentration in the liver (C.sub.max) and time to reach it (T.sub.max), half-life (T.sub.1/2), the area under the PC curve (AUC.sub.0-t, AUC.sub.0-inf), clearance (Cl), and elimination constant (kel).
TABLE-US-00004 TABLE 3 Pharmacokinetics of compound 1.1.19 and CAB in rat plasma when administered intravenously (IV) and orally (Oral). Unit of IV (dose: 1 mg/kg) measure- CAB Oral (dose: 5 mg/kg) Parameter ment 1.1.19 Cabotegravir 1.1.19 CAB AUC.sub.INF h .Math. ng/ml 367627 232346 282070 158201 AUC.sub.last h .Math. ng/ml 202189 140844 176506 62277 C.sub.max ng/ml 18550 16983 11488 5213 C.sub.8 h ng/ml 7913 4338 C.sub.24 h ng/ml 4435 1845 kel 1/h 0.03 0.04 0.04 0.04 MRT.sub.last h 9.46 9.19 9.42 8.05 T.sub.1/2 h 21.2 18.3 16.5 19.1 T.sub.max h 0.08 0.08 3 4.67 F % 17.5 8.8
Example 13. Evaluation of Stability in Biological Media of Compositions Comprising Compounds of General Formulas 1 and 2
[0301] a) Stability in blood plasma. A test compound at a final concentration of 1 μM was incubated in pooled human blood plasma (Innovative Research). Incubation was carried out in a VorTemp 56 shaking incubator at 37° C. with stirring at 300 rpm. At certain intervals (0, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 h30 μl of samples were taken, and the reaction was stopped by adding 180 μl of cold acetonitrile containing an internal standard to the selected sample. The samples were then centrifuged for 10 min at 3000 rpm, and 150 μl of the supernatant was taken for analysis. Incubation was performed in duplicate, each sample was measured twice. Samples were analyzed by HPLC-MS/MS developed for each prodrug tested using a 1290 Infinity II chromatographic system (Agilent Technologies) coupled to a QTRAP5500 tandem mass spectrometer (AB Sciex). When developing conditions for mass spectrometric detection, the solutions of tested compounds in a 1:1 acetonitrile-water mixture with a concentration of 100 ng/ml were analyzed by direct injection into the mass spectrometer by a syringe pump using electrospray ionization in positive ion mode. When scanning in the total ion current mode (MS1), the molecular ion for each compound was determined, the main product ions were recorded in the MS2 mode. Then, the MS/MS technique was optimized in the MRM mode to achieve maximum sensitivity. In the quantitative processing of chromatograms, the most intense MRM transition was used for the analyte and the internal standard. Chromatographic separation was performed on a YMC Triart C18, 50×2.mm, 1.9 μm column in a gradient elution mode in the mobile phase with a composition of 0.1% formic acid in water 0.1% formic acid in acetonitrile. Tolbutamide (Fluka) was used as an internal standard. The half-life (T.sub.1/2) was calculated from the kinetics of the loss of the tested prodrug in the antiviral composition during incubation in the biological medium. In calculations, the values of the areas of chromatographic peaks of substances in the experimental samples normalized to the signal of the internal standard were used. The elimination rate constant (k is the slope of the linear section) was calculated from the linear dependence of the log-normalized areas of chromatographic peaks on time. Then, the half-life was calculated: T.sub.1/2=0.693/k. It was found (
[0302] b) Stability in the S9 fraction. The reaction mixture was prepared in 0.1 M potassium phosphate buffer (pH 7.4 BD Gentest) in a total final volume of 250 μl and contained 1 mM of NADPH tetrasodium salt (AppliChem), 7 mM of glucose-6-phosphate sodium salt (Sigma), 1.5 U/ml of glucose-6-phosphate dehydrogenase (Sigma), 3.3 mM of MgCl.sub.2 (Sigma), 5 mM of uridine-5-diphosphate-glucuronic acid trisodium salt (UDPGA, Sigma), and 1 μM of the test compound. The metabolic reaction was initiated by adding a suspension of the S9 human (BD Gentest) or rat liver fraction, the final protein concentration was 1 mg/ml. The reaction mixture was incubated at 37° C. on a shaker (Vortemp56) with stirring at 400 rpm. At certain time intervals (0, 0.25, 0.5, 1, 2, 4, 6, 8, 24 h), 30-1 samples were taken, and the reaction was stopped by adding to the selected sample 180 μl of cold acetonitrile containing the internal standard. Proteins were precipitated on ice for 15 min, the samples were centrifuged for 10 min at 3000 rpm, and 150 μl of the supernatant was collected for analysis. Incubation was performed in duplicate with measuring each sample twice. It was established (
Example 14. Plasma Protein Binding of Compounds 1.1.17 and 1.1.19
[0303] To assess protein binding, pooled samples of blood plasma diluted with phosphate buffer up to 50% were used. The test was carried out in a teflon coated 48-well dialysis plate. Each well contained two separate chambers separated by a vertical semipermeable dialysis membrane with 8 kDa pores. A plasma sample with 1 μM of the test compound was placed into one of the chambers, and a buffer solution (pH=7.2) was placed in the other chamber. Over time, at 37° C. while rocking, passive diffusion of the unbound compound occurred and after 4 hours the equilibrium state between the chambers with plasma and the buffer solution was reached. The amount of free fraction was evaluated by HPLC-MS/MS following precipitation of the proteins by acetonitrile. Also, the study involved evaluation of stability of the substance during the 4-hour period and passive permeability through the dialysis membrane in the buffer by mass balance. Warfarin was used as the control compound. The results are presented in Table 4, from which it follows that the compounds of general formula 1.1 have a high degree of binding to human and rat plasma proteins.
TABLE-US-00005 TABLE 4 Binding of compounds 1.1.17 and 1.1.19 to human and rat plasma proteins Permeability, Extraction, Bound, Binding Type Compound % % % class Human 1.1.17 84.8 94.6 98.8 High Rat 1.1.17 90.9 108 99.9 High Human 1.1.19 84.0 95.0 99.0 High Rat 1.1.19 83.8 94.5 98.9 High
INDUSTRIAL APPLICABILITY
[0304] This invention can be applied in human and veterinary medicine.