6,7-dihydropyrido[2,1-A]phthalazin-2-ones for the treatment and prophylaxis of hepatitis B virus infection

Abstract

The invention provides novel compounds having the general formula: ##STR00001##
wherein R.sup.1 to R.sup.6 are as described herein, compositions including the compounds and methods of using the compounds.

Claims

1. A compound of formula I, ##STR00051## wherein R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are independently selected from C.sub.1-6alkyl, haloC.sub.1-6alkyl, hydrogen, halogen, amino, cyano, pyrrolidinyl, hydroxy, C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, C.sub.3-7cycloalkylC.sub.1-6alkyloxy, phenylC.sub.1-6alkyloxy, hydroxyC.sub.1-6alkyloxy, C.sub.1-6alkoxyC.sub.1-6alkyloxy, C.sub.1-6alkylsulfanylC.sub.1-6alkyloxy, C.sub.1-6alkylsulfonylC.sub.1-6alkyloxy, cyanoC.sub.1-6alkyloxy, aminoC.sub.1-6alkyloxy, C.sub.1-6alkylaminoC.sub.1-6alkyloxy, diC.sub.1-6alkylaminoC.sub.1-6alkyloxy, C.sub.1-6alkylcarbonylaminoC.sub.1-6alkyloxy, C.sub.1-6alkylsulfonylaminoC.sub.1-6alkyloxy, C.sub.1-6alkoxycarbonylaminoC.sub.1-6alkyloxy, pyrazolylC.sub.1-6alkyloxy, triazolylC.sub.1-6alkyloxy and monocyclic heterocycloalkylC.sub.1-6alkyloxy, wherein monocyclic heterocycloalkyl is N-containing monocyclic heterocycloalkyl; R.sup.5 and R.sup.6 are independently selected from hydrogen, C.sub.1-6alkyl, haloC.sub.1-6alkyl or C.sub.2-6alkenyl; or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

2. The compound of formula I according to claim 1, wherein R.sup.1 is hydrogen; R.sup.2 is C.sub.1-6alkoxy; R.sup.3 is C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, hydroxyC.sub.1-6alkoxy, C.sub.1-6alkoxyC.sub.1-6alkyloxy, C.sub.1-6alkylsulfonylC.sub.1-6alkoxy, cyanoC.sub.1-6alkoxy, carboxyC.sub.1-6alkoxy, aminoC.sub.1-6alkoxy, C.sub.1-6alkoxycarbonylaminoC.sub.1-6alkoxy, morpholinylC.sub.1-6alkoxy or tetrahydropyranylC.sub.1-6alkoxy; R.sup.4 is hydrogen; R.sup.5 is hydrogen; R.sup.6 is hydrogen, C.sub.1-6alkyl or C.sub.2-6alkenyl; or a pharmaceutically acceptable salt, or an enantiomer, or diastereomer thereof.

3. A compound of formula I according to claim 2, wherein R.sup.2 is methoxy; R.sup.3 methoxy, trifluoroethoxy, hydroxydimethylpropoxy, methoxypropoxy, methylsulfonylpropoxy, cyanopropoxy, carboxybutoxy, aminopentoxy, tert-butoxycarbonylaminopentoxy, morpholinylethoxy or tetrahydropyranylmethoxy; R.sup.6 is hydrogen, methyl, isopropyl, isobutyl or prop-2-enyl; or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

4. The compound of formula I according to claim 1, or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof, wherein R.sup.2 is methoxy.

5. The compound of formula I according to claim 1, or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof, wherein R.sup.3 is haloC.sub.1-6alkoxy, hydroxyC.sub.1-6alkoxy, C.sub.1-6alkoxyC.sub.1-6alkyloxy, cyanoC.sub.1-6alkoxy, C.sub.1-6alkoxycarbonylaminoC.sub.1-6alkoxy or tetrahydropyranylC.sub.1-6alkoxy.

6. The compound of formula I according to claim 5, or a pharmaceutically acceptable salt, or an enantiomers, or a diastereomer thereof, wherein R.sup.3 is trifluoroethoxy, hydroxydimethylpropoxy, methoxypropoxy, cyanopropoxy, tert-butoxycarbonylaminopentoxy, or tetrahydropyranylmethoxy.

7. The compound of formula I according to claim 1, or pharmaceutically acceptable salts, or enantiomers, or diastereomer thereof, wherein R.sup.6 is C.sub.1-6alkyl.

8. The compound of formula I according to claim 7, or a pharmaceutically acceptable salt, or an enantiomers, or a diastereomer thereof, wherein R.sup.6 is isobutyl.

9. The compound according to claim 1, which compound is selected from the group consisting of: 9,10-Dimethoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylic acid; 9,10-Dimethoxy-6-methyl-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Allyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isopropyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isobutyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isobutyl-10-methoxy-9-(3-methoxypropoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isobutyl-10-methoxy-2-oxo-9-(2,2,2-trifluoroethoxy)-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isobutyl-10-methoxy-2-oxo-9-(tetrahydropyran-4-ylmethoxy)-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isobutyl-10-methoxy-9-(3-methylsulfonylpropoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isobutyl-10-methoxy-9-(2-morpholinoethoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 9-(3-Hydroxy-2,2-dimethyl-propoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 9-(4-Carboxybutoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 9-(3-Cyanopropoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 9-[5-(Tert-butoxycarbonylamino)pentoxy]-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; and, 9-(5-Aminopentoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid hydrochloride; or, a pharmaceutically acceptable salt, or an enantiomers, or a diastereomer thereof.

10. The compound according to claim 9, which compound is selected from the group consisting of: 6-Isobutyl-10-methoxy-9-(3-methoxypropoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isobutyl-10-methoxy-2-oxo-9-(tetrahydropyran-4-ylmethoxy)-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 9-(3-Hydroxy-2,2-dimethyl-propoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; and, 9-[5-(Tert-butoxycarbonylamino)pentoxy]-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; or, a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

11. A process for the preparation of a compound according to claim 1 comprising hydrolysis of a compound of formula (A) to afford a compound of formula (I) ##STR00052## wherein R.sup.11 is C.sub.1-6 alkyl.

12. The process of according to claim 11 wherein R.sup.3 is C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, C.sub.3-7cycloalkylC.sub.1-6alkyloxy, phenylC.sub.1-6alkyloxy, hydroxyC.sub.1-6alkyloxy, C.sub.1-6alkoxyC.sub.1-6alkyloxy, C.sub.1-6alkylsulfanylC.sub.1-6alkyloxy, C.sub.1-6alkylsulfonylC.sub.1-6alkyloxy, cyanoC.sub.1-6alkyloxy, aminoC.sub.1-6alkyloxy, C.sub.1-6alkylaminoC.sub.1-6alkyloxy, diC.sub.1-6alkylaminoC.sub.1-6alkyloxy, C.sub.1-6alkylcarbonylaminoC.sub.1-6alkyloxy, C.sub.1-6alkylsulfonylaminoC.sub.1-6alkyloxy, C.sub.1-6alkoxycarbonylaminoC.sub.1-6alkyloxy, pyrazolylC.sub.1-6alkyloxy, triazolylC.sub.1-6alkyloxy and monocyclic heterocycloalkylC.sub.1-6alkyloxy, wherein monocyclic heterocycloalkyl is N-containing monocyclic heterocycloalkyl.

13. A pharmaceutical composition comprising a compound in accord with claim 1 and at least one pharmaceutically acceptable carrier, diluent or excipient.

14. The method of inhibiting HBsAg production or secretionin a cell comprising treating the cell with a therapeutically effective amount of a compound according to claim 1.

15. A method for the treatment or prophylaxis of HBV infection, which method comprises administering to a patient in need thereof a therapeutically effective amount of a compound according to claim 1.

Description

DETAILED DESCRIPTION OF THE INVENTION

(1) Definitions

(2) As used herein, the term C.sub.1-6alkyl alone or in combination signifies a saturated, linear- or branched chain alkyl group containing 1 to 6, particularly 1 to 4 carbon atoms, for example methyl, ethyl, propyl, isopropyl, 1-butyl, 2-butyl, tert-butyl and the like. Particular C.sub.1-6alkyl groups are methyl, ethyl, isopropyl and tert-butyl.

(3) The term C.sub.2-6alkenyl denotes a monovalent linear or branched hydrocarbon group of 2 to 6 carbon atoms with at least one double bond. In particular embodiments, C.sub.1-6alkenyl has 2 to 4 carbon atoms with at least one double bond. Examples of alkenyl include ethenyl, propenyl, prop-2-enyl, isopropenyl, n-butenyl, and iso-butenyl. Particular C.sub.1-6alkyl group is prop-2-enyl.

(4) The term C.sub.3-7cycloalkyl, alone or in combination, refers to a saturated carbon ring containing from 3 to 7 carbon atoms, particularly from 3 to 6 carbon atoms, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like. Particular C.sub.3-7cycloalkyl groups are cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

(5) The term C.sub.1-6alkoxy alone or in combination signifies a group C.sub.1-6alkyl-O, wherein the C.sub.1-6alkyl is as defined above; for example methoxy, ethoxy, propoxy, iso-propoxy, n-butoxy, iso-butoxy, 2-butoxy, tert-butoxy, pentoxy, hexyloxy, octyloxy and the like. Particular C.sub.1-6alkoxy groups are methoxy, ethoxy and propoxy.

(6) The term halogen means fluorine, chlorine, bromine or iodine.

(7) The term haloC.sub.1-6alkyl denotes a C.sub.1-6alkyl group wherein at least one of the hydrogen atoms of the C.sub.1-6alkyl group has been replaced by same or different halogen atoms, particularly fluoro atoms. Examples of haloC.sub.1-6alkyl include monofluoro-, difluoro- or trifluoro-methyl, -ethyl or -propyl, for example 3,3,3-trifluoropropyl, 3,3-difluoropropyl, 2-fluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, fluoromethyl, difluoromethyl or trifluoromethyl. Particular haloC.sub.1-6alkyl group is difluoromethyl or trifluoromethyl.

(8) The term haloC.sub.1-6alkoxy denotes a C.sub.1-6alkoxy group wherein at least one of the hydrogen atoms of the C.sub.1-6alkoxy group has been replaced by same or different halogen atoms, particularly fluoro atoms. Examples of haloC.sub.1-6alkoxyl include monofluoro-, difluoro- or trifluoro-methoxy, -ethoxy or -propoxy, for example fluoropropoxy, difluoropropoxy, trifluoropropoxy, fluoroethoxy, difluoroethoxy, trifluoroethoxy, fluoromethoxy, difluoromethoxy or trifluoromethoxy. Particular haloC.sub.1-6alkoxy group is 3-fluoropropoxy, 3,3-difluoropropoxy, 3,3,3-trifluoropropoxy, 2-fluoroethoxy, 2,2-difluoroethoxy, 2,2,2-trifluoroethoxy, fluoromethoxy, difluoromethoxy or trifluoromethoxy.

(9) The term amino denotes a group of the formula NRR wherein R and R are independently hydrogen, C.sub.1-6alkyl, C.sub.1-6alkoxy, C.sub.3-7cycloalkyl, heteroC.sub.3-7cycloalkyl, aryl or heteroaryl. Alternatively, R and R, together with the nitrogen to which they are attached, can form a heteroC.sub.3-7cycloalkyl.

(10) The term carbonyl alone or in combination refers to the group C(O).

(11) The term cyano alone or in combination refers to the group CN.

(12) The term C.sub.1-6alkylsulfanyl denotes the group SR, wherein R is a C.sub.1-6alkyl group as defined above. Examples of C.sub.1-6alkylsulfanyl include methylsulfanyl and ethylsulfanyl.

(13) The term C.sub.1-6alkylsulfonyl denotes a group SO.sub.2R, wherein R is a C.sub.1-6alkyl group as defined above. Examples of C.sub.1-6alkylsulfonyl include methylsulfonyl and ethylsulfonyl.

(14) The term monocyclic heterocycloalkyl is a monovalent saturated or partly unsaturated monocyclic ring system of 4 to 7 ring atoms, comprising 1, 2, or 3 ring heteroatoms selected from N, O and S, the remaining ring atoms being carbon. Examples for monocyclic heterocycloalkyl are aziridinyl, oxiranyl, azetidinyl, oxetanyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, tetrahydrofuranyl, tetrahydro-thienyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperazinyl, morpholinyl, 2-oxo-morpholinyl, 2-oxo-piperazinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholin-4-yl, 1,1-dioxothiolanyl, azepanyl, diazepanyl, homopiperazinyl, or oxazepanyl. Particular monocyclic heterocycloalkyl groups are oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,1-dioxothiolanyl, morpholinyl, 2-oxo-pyrrolidinyl, 2-oxo-morpholinyl and 2-oxo-piperazinyl.

(15) The term N-containing monocyclic heterocycloalkyl refers to a monocyclic heterocycloalkyl wherein at least one of the heteroatoms is N. Examples for N-containing monocyclic heterocycloalkyl are azetidinyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, piperidinyl, piperazinyl, morpholinyl, azepanyl, diazepanyl, homopiperazinyl or oxazepanyl. Particular N-containing monocyclic heterocycloalkyl groups are pyrrolidinyl, 2-oxo-pyrrolidinyl, piperidinyl, piperazinyl, tetrahydropyranyl and morpholinyl, and more particularly tetrahydropyranyl and morpholinyl.

(16) The term enantiomer denotes two stereoisomers of a compound which are non-superimposable mirror images of one another.

(17) The term diastereomer denotes a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical properties, e.g. melting points, boiling points, spectral properties, and reactivities.

(18) The compounds according to the present invention may exist in the form of their pharmaceutically acceptable salts. The term pharmaceutically acceptable salt refers to conventional acid-addition salts or base-addition salts that retain the biological effectiveness and properties of the compounds of formula I and are formed from suitable non-toxic organic or inorganic acids or organic or inorganic bases. Acid-addition salts include for example those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, phosphoric acid and nitric acid, and those derived from organic acids such as p-toluenesulfonic acid, salicylic acid, methanesulfonic acid, oxalic acid, succinic acid, citric acid, malic acid, lactic acid, fumaric acid, and the like. Base-addition salts include those derived from ammonium, potassium, sodium and, quaternary ammonium hydroxides, such as for example, tetramethyl ammonium hydroxide. The chemical modification of a pharmaceutical compound into a salt is a technique well known to pharmaceutical chemists in order to obtain improved physical and chemical stability, hygroscopicity, flowability and solubility of compounds. It is for example described in Bastin R. J., et al., Organic Process Research & Development 2000, 4, 427-435. Particular are the sodium salts of the compounds of formula I.

(19) Compounds of the general formula I which contain one or several chiral centers can either be present as racemates, diastereomeric mixtures, or optically active single isomers. The racemates can be separated according to known methods into the enantiomers. Particularly, diastereomeric salts which can be separated by crystallization are formed from the racemic mixtures by reaction with an optically active acid such as e.g. D- or L-tartaric acid, mandelic acid, malic acid, lactic acid or camphorsulfonic acid.

(20) Inhibitor of HBsAg

(21) The present invention provides (i) a compound having the general formula I:

(22) ##STR00004##
wherein
R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are independently selected from C.sub.1-6alkyl, haloC.sub.1-6alkyl, hydrogen, halogen, amino, cyano, pyrrolidinyl, hydroxy, C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, C.sub.3-7cycloalkylC.sub.1-6alkyloxy, phenylC.sub.1-6alkyloxy, hydroxyC.sub.1-6alkyloxy, C.sub.1-6alkoxyC.sub.1-6alkyloxy, C.sub.1-6alkylsulfanylC.sub.1-6alkyloxy, C.sub.1-6alkylsulfonylC.sub.1-6alkyloxy, cyanoC.sub.1-6alkyloxy, aminoC.sub.1-6alkyloxy, C.sub.1-6alkylaminoC.sub.1-6alkyloxy, diC.sub.1-6alkylaminoC.sub.1-6alkyloxy, C.sub.1-6alkylcarbonylaminoC.sub.1-6alkyloxy, C.sub.1-6alkylsulfonylaminoC.sub.1-6alkyloxy, C.sub.1-6alkoxycarbonylaminoC.sub.1-6alkyloxy, pyrazolylC.sub.1-6alkyloxy, triazolylC.sub.1-6alkyloxy and monocyclic heterocycloalkylC.sub.1-6alkyloxy, wherein monocyclic heterocycloalkyl is N-containing monocyclic heterocycloalkyl;
R.sup.5 and R.sup.6 are independently selected from hydrogen, C.sub.1-6alkyl, haloC.sub.1-6alkyl or C.sub.2-6alkenyl; or pharmaceutically acceptable salts, or enantiomers, or diastereomer thereof.

(23) Another embodiment of present invention is (ii) a compound of formula I, wherein

(24) R.sup.1 is hydrogen;

(25) R.sup.2 is C.sub.1-6alkoxy;

(26) R.sup.3 is C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, hydroxyC.sub.1-6alkoxy, C.sub.1-6alkoxyC.sub.1-6alkoxy, C.sub.1-6alkylsulfonylC.sub.1-6alkoxy, cyanoC.sub.1-6alkoxy, carboxyC.sub.1-6alkoxy, aminoC.sub.1-6alkoxy, C.sub.1-6alkoxycarbonylaminoC.sub.1-6alkoxy, morpholinylC.sub.1-6alkoxy or tetrahydropyranylC.sub.1-6alkoxy;

(27) R.sup.4 is hydrogen;

(28) R.sup.5 is hydrogen;

(29) R.sup.6 is hydrogen, C.sub.1-6alkyl or C.sub.2-6alkenyl;

(30) or pharmaceutically acceptable salts, or enantiomers, or diastereomer thereof.

(31) A further embodiment of present invention is (iii) a compound of formula I, wherein

(32) R.sup.1 is hydrogen;

(33) R.sup.2 is methoxy;

(34) R.sup.3 methoxy, trifluoroethoxy, hydroxydimethylpropoxy, methoxypropoxy, methylsulfonylpropoxy, cyanopropoxy, carboxybutoxy, aminopentoxy, tert-butoxycarbonylaminopentoxy, morpholinylethoxy or tetrahydropyranylmethoxy;

(35) R.sup.4 is hydrogen;

(36) R.sup.5 is hydrogen;

(37) R.sup.6 is hydrogen, methyl, isopropyl, isobutyl or prop-2-enyl;

(38) or pharmaceutically acceptable salts, or enantiomers, or diastereomer thereof.

(39) Another embodiment of present invention is (iv) a compound of formula I or pharmaceutically acceptable salts, or enantiomers, or diastereomer thereof, wherein R.sup.2 is methoxy, and all remaining substituents have the significances given herein before.

(40) Another embodiment of present invention is (v) a compound of formula I or pharmaceutically acceptable salts, or enantiomers, or diastereomer thereof, wherein R.sup.3 is haloC.sub.1-6alkoxy, hydroxyC.sub.1-6alkoxy, C.sub.1-6alkoxyC.sub.1-6alkoxy, cyanoC.sub.1-6alkoxy, C.sub.1-6alkoxycarbonylaminoC.sub.1-6alkoxy or tetrahydropyranylC.sub.1-6alkoxy, and all remaining substituents have the significances given herein before.

(41) A further embodiment of present invention is (vi) a compound of formula I or pharmaceutically acceptable salts, or enantiomers, or diastereomer thereof, wherein R.sup.3 is trifluoroethoxy, hydroxydimethylpropoxy, methoxypropoxy, cyanopropoxy, tert-butoxycarbonylaminopentoxy, or tetrahydropyranylmethoxy, and all remaining substituents have the significances given herein before.

(42) Another embodiment of present invention is (vii) a compound of formula I or pharmaceutically acceptable salts, or enantiomers, or diastereomer thereof, wherein R.sup.6 is C.sub.1-6alkyl, and all remaining substituents have the significances given herein before.

(43) A further embodiment of present invention is (viii) a compound of formula I or pharmaceutically acceptable salts, or enantiomers, or diastereomer thereof, wherein R.sup.6 is isobutyl, and all remaining substituents have the significances given herein before.

(44) Particular compounds of formula I according to the invention are the following: 9,10-Dimethoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylic acid; 9,10-Dimethoxy-6-methyl-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Allyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isopropyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isobutyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isobutyl-10-methoxy-9-(3-methoxypropoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isobutyl-10-methoxy-2-oxo-9-(2,2,2-trifluoroethoxy)-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isobutyl-10-methoxy-2-oxo-9-(tetrahydropyran-4-ylmethoxy)-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isobutyl-10-methoxy-9-(3-methylsulfonylpropoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isobutyl-10-methoxy-9-(2-morpholinoethoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 9-(3-Hydroxy-2,2-dimethyl-propoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 9-(4-Carboxybutoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 9-(3-Cyanopropoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 9-[5-(Tert-butoxycarbonylamino)pentoxy]-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 9-(5-Aminopentoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid hydrochloride
or pharmaceutically acceptable salts, or enantiomers, or diastereomer thereof.
10. A compound according to claim 1, selected from 6-Isobutyl-10-methoxy-9-(3-methoxypropoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isobutyl-10-methoxy-2-oxo-9-(tetrahydropyran-4-ylmethoxy)-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 9-(3-Hydroxy-2,2-dimethyl-propoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 9-[5-(Tert-butoxycarbonylamino)pentoxy]-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid;
or pharmaceutically acceptable salts, or enantiomers, or diastereomer thereof.

(45) More particularly, the invention relates to the following compounds of formula I: 6-Isobutyl-10-methoxy-9-(3-methoxypropoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 6-Isobutyl-10-methoxy-2-oxo-9-(tetrahydropyran-4-ylmethoxy)-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 9-(3-Hydroxy-2,2-dimethyl-propoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid; 9-[5-(Tert-butoxycarbonylamino)pentoxy]-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid;
or pharmaceutically acceptable salts, or enantiomers, or diastereomer thereof.
Synthesis

(46) The compounds of the present invention can be prepared by any conventional means. Suitable processes for synthesizing these compounds as well as their starting materials are provided in the schemes below and in the examples. All substituents, in particular, R.sup.1 to R.sup.6 are as defined above unless otherwise indicated. Furthermore, and unless explicitly otherwise stated, all reactions, reaction conditions, abbreviations and symbols have the meanings well known to a person of ordinary skill in organic chemistry.

General Synthetic Route for Intermediates (Scheme 1)

(47) ##STR00005##

(48) The intermediate of formula V can be prepared according to Scheme 1. Compound II reacts with DMFDMA at a temperature between 0 and 80 C. to afford compound III, which reacts with ethyl formate in the presence of a base such as t-BuOK, and then the resulting reaction mixture is treated with an acid such as hydrochloric acid to afford compound IV. Compound IV reacts with tert-butyl N-aminocarbamate in a suitable solvent such as EtOH at a temperature between rt and 100 C. to give compound V.

General Synthetic Route for Compounds I (Scheme 2)

(49) ##STR00006##

(50) The compound of formula I can be prepared according to Scheme 2, wherein X is halogen; Y is halogen or triflate; Q.sup.1 is halogen, OS(O).sub.2CH.sub.3 or OS(O).sub.2-(4-methylphenyl); R.sup.11 is C.sub.1-6alkyl. Reaction of substituted benzene VI with compound V in the presence of a base such as K.sub.2CO.sub.3 affords Compound VII, which undergoes intramolecular coupling reaction to give compound VIII. The coupling reaction can be carried out in the presence of a Pd catalyst such as PdBr.sub.2 and a suitable base such as KOAc in a suitable solvent such as DMA at a temperature between room temperature and 150 C. Compound VIII reacts with halide, mesylate or tosylate in the presence of a base such as K.sub.2CO.sub.3 in a solvent such as acetonitrile or DMF to give compound VIII-1. Hydrolyzation of Compound VIII-1 with an acid such as TFA or HCl in a suitable solvent such as DCM, or with a base such as lithium hydroxide in a suitable solvent such as EtOH/H.sub.2O or MeOH/H.sub.2O affords the compound of formula I.

General Synthetic Route for Compounds I-1 (Scheme 3)

(51) ##STR00007##

(52) The compound of formula I-1 can be prepared according to Scheme 3, wherein Q.sup.1 is halogen, OS(O).sub.2CH.sub.3 or OS(O).sub.2-(4-methylphenyl); R.sup.8 is C.sub.1-6alkyl, haloC.sub.1-6alkyl, C.sub.3-7cycloalkylC.sub.1-6alkyl, phenylC.sub.1-6alkyl, hydroxyC.sub.1-6alkyl, C.sub.1-6alkoxyC.sub.1-6alkyl, C.sub.1-6alkylsulfanylC.sub.1-6alkyl, C.sub.1-6alkylsulfonylC.sub.1-6alkyl, cyanoC.sub.1-6alkyl, amino C.sub.1-6alkyl, C.sub.1-6alkylaminoC.sub.1-6alkyl, diC.sub.1-6alkylaminoC.sub.1-6alkyl, C.sub.1-6alkylcarbonylaminoC.sub.1-6alkyl, C.sub.1-6alkylsulfonylaminoC.sub.1-6alkyl, C.sub.1-6alkoxycarbonylaminoC.sub.1-6alkyl, pyrazolylC.sub.1-6alkyl, triazolylC.sub.1-6alkyl or monocyclic heterocycloalkylC.sub.1-6alkyl, wherein monocyclic heterocycloalkyl is N-containing monocyclic heterocycloalkyl; R.sup.11 is C.sub.1-6alkyl. Debenzylation of Compound VIII-2 by hydrogenation is carried out in the presence of Pd/C in a solvent such as ethanol to afford Compound VIII-3. Then Compound VIII-3 reacts with halide, mesylate or tosylate in the presence of a base such as K.sub.2CO.sub.3 in a solvent such as DMF to give VIII-4. Hydrolyzation of Compound VIII-4 with an acid such as TFA or HCl in a suitable solvent such as DCM, or with a base such as lithium hydroxide in a suitable solvent such as EtOH/H.sub.2O or MeOH/H.sub.2O affords the compound of formula I.

(53) This invention also relates to a process for the preparation of a compound of formula I comprising

(54) (a) hydrolysis of a compound of formula (A) by using an acid or a base

(55) ##STR00008##
or

(56) (b) hydrolysis of a compound of formula (B) by using an acid or a base

(57) ##STR00009##

(58) wherein R.sup.1 to R.sup.6, R.sup.8 and R.sup.11 are defined as above unless otherwise indicated.

(59) In step (a) and (b), an acid is for example TFA or HCl; or a base is for example lithium hydroxide or sodium hydroxide.

(60) A compound of formula I when manufactured according to the above process is also an object of the invention.

(61) The invention also relates to a compound of formula I for use as therapeutically active substance.

(62) Another embodiment provides pharmaceutical compositions or medicaments containing the compounds of the invention and a therapeutically inert carrier, diluent or excipient, as well as methods of using the compounds of the invention to prepare such compositions and medicaments. In one example, compounds of formula (I) may be formulated by mixing at ambient temperature at the appropriate pH, and at the desired degree of purity, with physiologically acceptable carriers, i.e., carriers that are non-toxic to recipients at the dosages and concentrations employed into a galenical administration form. The pH of the formulation depends mainly on the particular use and the concentration of compound, but preferably ranges anywhere from about 3 to about 8. In one example, a compound of formula (I) is formulated in an acetate buffer, at pH 5. In another embodiment, the compounds of formula (I) are sterile. The compound may be stored, for example, as a solid or amorphous composition, as a lyophilized formulation or as an aqueous solution.

(63) Compositions are formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners. The effective amount of the compound to be administered will be governed by such considerations, and is the minimum amount necessary to inhibit HBsAg. For example, such amount may be below the amount that is toxic to normal cells, or the mammal as a whole.

(64) In one example, the pharmaceutically effective amount of the compound of the invention administered parenterally per dose will be in the range of about 0.01 to 100 mg/kg, alternatively about 0.01 to 100 mg/kg of patient body weight per day, with the typical initial range of compound used being 0.3 to 15 mg/kg/day. In another embodiment, oral unit dosage forms, such as tablets and capsules, preferably contain from about 0.1 to about 1000 mg of the compound of the invention.

(65) The compounds of the invention may be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and epidural and intranasal, and, if desired for local treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.

(66) The compounds of the present invention may be administered in any convenient administrative form, e.g., tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain components conventional in pharmaceutical preparations, e.g., diluents, carriers, pH modifiers, sweeteners, bulking agents, and further active agents.

(67) A typical formulation is prepared by mixing a compound of the present invention and a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art. The formulations may also include one or more buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents, diluents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present invention or pharmaceutical composition thereof) or aid in the manufacturing of the pharmaceutical product (i.e., medicament).

(68) An example of a suitable oral dosage form is a tablet containing about 0.1 to 1000 mg of the compound of the invention compounded with about 0 to 2000 mg anhydrous lactose, about 0 to 2000 mg sodium croscarmellose, about 0 to 2000 mg polyvinylpyrrolidone (PVP) K30, and about 0 to 2000 mg magnesium stearate. The powdered ingredients are first mixed together and then mixed with a solution of the PVP. The resulting composition can be dried, granulated, mixed with the magnesium stearate and compressed to tablet form using conventional equipment. An example of an aerosol formulation can be prepared by dissolving the compound, for example 0.1 to 1000 mg, of the invention in a suitable buffer solution, e.g. a phosphate buffer, adding a tonicifier, e.g. a salt such sodium chloride, if desired. The solution may be filtered, e.g., using a 0.2 micron filter, to remove impurities and contaminants.

(69) An embodiment, therefore, includes a pharmaceutical composition comprising a compound of Formula I, or a stereoisomer or pharmaceutically acceptable salt thereof. In a further embodiment includes a pharmaceutical composition comprising a compound of Formula I, or a stereoisomer or pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier or excipient.

(70) The following example A and B illustrate typical compositions of the present invention, but serve merely as representative thereof.

Example A

(71) A compound of formula I can be used in a manner known per se as the active ingredient for the production of tablets of the following composition:

(72) TABLE-US-00001 Per tablet Active ingredient 200 mg Microcrystalline cellulose 155 mg Corn starch 25 mg Talc 25 mg Hydroxypropylmethylcellulose 20 mg 425 mg

Example B

(73) A compound of formula I can be used in a manner known per se as the active ingredient for the production of capsules of the following composition:

(74) TABLE-US-00002 Per capsule Active ingredient 100.0 mg Corn starch 20.0 mg Lactose 95.0 mg Talc 4.5 mg Magnesium stearate 0.5 mg 220.0 mg
Indications and Methods of Treatment

(75) The compounds of the invention can inhibit HBsAg production or secretion and inhibit HBV gene expression. Accordingly, the compounds of the invention are useful for the treatment or prophylaxis of HBV infection.

(76) The invention relates to the use of a compound of formula I for the inhibition of HBsAg production or secretion.

(77) The invention relates to the use of a compound of formula I for the inhibition of HBV DNA production.

(78) The invention relates to the use of a compound of formula I for the inhibition of HBV gene expression.

(79) The invention relates to the use of a compound of formula I for the treatment or prophylaxis of HBV infection.

(80) The use of a compound of formula I for the preparation of medicaments useful in the treatment or prophylaxis diseases that are related to HBV infection is an object of the invention.

(81) The invention relates in particular to the use of a compound of formula I for the preparation of a medicament for the treatment or prophylaxis of HBV infection.

(82) Another embodiment includes a method for the treatment or prophylaxis of HBV infection, which method comprises administering an effective amount of a compound of Formula I, a stereoisomer, tautomer, prodrug, conjugates or pharmaceutically acceptable salt thereof.

EXAMPLES

(83) The invention will be more fully understood by reference to the following examples. They should not, however, be construed as limiting the scope of the invention.

(84) Abbreviations used herein are as follows:

(85) L: microliter

(86) m: micrometer

(87) M: micromoles per liter

(88) DMF: dimethylformamide

(89) DMFDMA: N,N-dimethylformamide dimethyl acetal

(90) DMA: N,N-Dimethylaniline

(91) DMSO-d6: deuterated dimethylsulfoxide

(92) EtOAc: ethyl acetate

(93) h or hr: hour

(94) hrs: hours

(95) IC.sub.50: the half maximal inhibitory concentration

(96) HPLC: high performance liquid chromatography

(97) LC/MS: Liquid chromatography/mass spectrometry

(98) METHANOL-d.sub.4: perdeuteromethanol

(99) M: molarity

(100) mg: milligram

(101) MHz: megahertz

(102) min: minute

(103) mL: milliliter

(104) mM: millimoles per liter

(105) mm: millimeter

(106) mmol: millimole

(107) MS (ESI): mass spectroscopy (electron spray ionization)

(108) NMR: nuclear magnetic resonance

(109) N.sub.2: nitrogen

(110) rt: room temperature

(111) Pd/C: palladium on activated carbon

(112) prep-HPLC: preparative high performance liquid chromatography

(113) TFA: trifluoroacetic acid

(114) : chemical shift

(115) t-BuOK: potassium tert-butylate

(116) General Experimental Conditions

(117) Intermediates and final compounds were purified by flash chromatography using one of the following instruments: i) Biotage SP1 system and the Quad 12/25 Cartridge module. ii) ISCO combi-flash chromatography instrument. Silica gel Brand and pore size: i) KP-SIL 60 , particle size: 40-60 m; ii) CAS registry NO: Silica Gel: 63231-67-4, particle size: 47-60 micron silica gel; iii) ZCX from Qingdao Haiyang Chemical Co., Ltd, pore: 200-300 or 300-400.

(118) Intermediates and final compounds were purified by preparative HPLC on reversed phase column using X Bridge Perp C.sub.18 (5 m, OBD 30100 mm) column or SunFire Perp C.sub.18 (5 m, OBD 30100 mm) column.

(119) LC/MS spectra were obtained using an Acquity Ultra Performance LC-3100 Mass Detector or Acquity Ultra Performance LC-SQ Detector. Standard LC/MS conditions were as follows (running time 3 minutes):

(120) Acidic condition: A: 0.1% formic acid in H.sub.2O; B: 0.1% formic acid in acetonitrile;

(121) Basic condition: A: 0.05% NH.sub.3.H.sub.2O in H.sub.2O; B: acetonitrile;

(122) Neutral condition: A: H.sub.2O; B: acetonitrile.

(123) Mass spectra (MS): generally only ions which indicate the parent mass are reported, and unless otherwise stated the mass ion quoted is the positive mass ion (M+H).sup.+.

(124) The microwave assisted reactions were carried out in a Biotage Initiator Sixty or CEM Discover.

(125) NMR Spectra were obtained using Bruker Avance 400 MHz.

(126) All reactions involving air-sensitive reagents were performed under an argon atmosphere. Reagents were used as received from commercial suppliers without further purification unless otherwise noted.

Preparative Examples

Example 1: 9,10-Dimethoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylic acid

(127) ##STR00010##

Step 1: Preparation of tert-butyl 2-(dimethylaminomethylene)-3-oxo-butanoate

(128) ##STR00011##

(129) To a stirred solution of tert-butyl 3-oxobutanoate (30.0 g, 0.19 mol) in 1,4-dioxane (500 mL) was added N,N-dimethylformamide dimethyl acetal (113.0 g, 0.95 mol). The mixture was stirred at 25 C. for 16 hrs, and then concentrated under reduced pressure. The residue was diluted with H.sub.2O (300 mL). The resulting mixture was extracted with EtOAc (200 mL) for three times. The combined organic layers were dried over anhydrous Na.sub.2SO.sub.4 and concentrated under reduced pressure to give crude tert-butyl 2-(dimethylaminomethylene)-3-oxo-butanoate (40.0 g) as a dark yellow liquid, which was used directly in the next step without further purification.

Step 2: Preparation of tert-butyl 4-oxopyran-3-carboxylate

(130) ##STR00012##

(131) To a stirred solution of tert-butyl 2-(dimethylaminomethylene)-3-oxo-butanoate (40.0 g, 0.19 mol) and ethyl formate (27.8 g, 0.36 mol) in THF (700 mL) was added t-BuOK (52.6 g, 0.47 mol) portion wise at 0 C. Then the mixture was allowed to warm to 25 C. and stirred at the same temperature for 16 hrs. Then the reaction was quenched by adding 1 M hydrochloric acid. The resulting mixture was extracted with EtOAc (200 mL) for three times. The combined organic layers were washed with saturated NaHCO.sub.3 aqueous solution and brine, dried over anhydrous Na.sub.2SO.sub.4, and concentrated under reduced pressure. The residue was purified by the flash column chromatography to give tert-butyl 4-oxopyran-3-carboxylate (9.5 g) as a dark yellow solid.

Step 3: Preparation of tert-butyl 1-ethyl-4-oxo-pyridine-3-carboxylate

(132) ##STR00013##

(133) A suspension of tert-butyl N-aminocarbamate (7.0 g, 53.0 mmol) and tert-butyl 4-oxopyran-3-carboxylate (6.9 g, 35.3 mmol) in absolute EtOH (70 mL) was heated under reflux with stirring for 48 hrs. Then the mixture was cooled to rt and concentrated to give crude tert-butyl 1-ethyl-4-oxo-pyridine-3-carboxylate (1.4 g) which was used in the next step without further purification.

Step 4: Preparation of 1-bromo-2-(chloromethyl)-4,5-dimethoxy-benzene

(134) ##STR00014##

(135) A mixture of (2-bromo-4,5-dimethoxy-phenyl)methanol (1.0 g, 4.05 mmol) and SOCl.sub.2 (0.32 mL, 4.46 mmol) in DCM (10 mL) was stirred at rt for 3 hrs. The reaction mixture was then concentrated to give 1-bromo-2-(chloromethyl)-4,5-dimethoxy-benzene (1.1 g) which was used in the next step without further purification.

Step 5: Preparation of tert-butyl 1-[2-(2-bromo-4,5-dimethoxy-phenyl)-1-methyl-ethyl]-4-oxo-pyridine-3-carboxylate

(136) ##STR00015##

(137) A mixture of tert-butyl 1-ethyl-4-oxo-pyridine-3-carboxylate (1.4 g, 4.52 mmol), 1-bromo-2-(chloromethyl)-4,5-dimethoxy-benzene (2.4 g, 9.03 mmol) and K.sub.2CO.sub.3 (1.3 g, 9.03 mmol) in acetone (20 mL) was heated at 50 C. with stirring for 2 hrs. Then the mixture was cooled to rt and diluted with water (20 mL). The resulting mixture was extracted with EtOAc (50 mL) for three times. The combined organic layers were washed with brine, dried over anhydrous Na.sub.2SO.sub.4, and concentrated under reduced pressure to give tert-butyl 1-[2-(2-bromo-4,5-dimethoxy-phenyl)-1-methyl-ethyl]-4-oxo-pyridine-3-carboxylate (2.6 g) which was used in the next step without further purification.

Step 6: Preparation of tert-butyl 9,10-dimethoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylate

(138) ##STR00016##

(139) A mixture of tert-butyl 1-[2-(2-bromo-4,5-dimethoxy-phenyl)-1-methyl-ethyl]-4-oxo-pyridine-3-carboxylate (4.0 g, 7.42 mmol), PdBr.sub.2 (99 mg, 0.37 mmol) and KOAc (1.45 g, 14.84 mmol) in DMA (15 mL) was heated at 120 C. with stirring overnight. Then the mixture was cooled to rt and filtered. The filtrate was concentrated under reduced pressure and then diluted with water (30 mL). The mixture was extracted with EtOAc (30 mL) for three times. The combined organic layers were washed with brine, dried over anhydrous Na.sub.2SO.sub.4, and concentrated to give crude tert-butyl 9,10-dimethoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylate (2.0 g) which was used in the next step without further purification.

Step 7: Preparation of 9,10-dimethoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylic acid

(140) ##STR00017##

(141) To a solution of tert-butyl 9,10-dimethoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylate (100 mg, 0.28 mmol) in a mixed solvent of THF (5 mL) and H.sub.2O (5 mL) was added LiOH.H.sub.2O (118 mg, 2.8 mmol). The mixture was heated at 90 C. with stirring for 3 hrs. The mixture was cooled to rt and extracted with EtOAc (20 mL) for three times. The combined organic layers were washed with brine, dried over anhydrous Na.sub.2SO.sub.4, and concentrated under reduced pressure. The residue was purified by prep-HPLC to give 9,10-dimethoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylic acid (10 mg) as a light yellow solid, .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 3.87 (d, 6H) 4.09-4.34 (m, 2H), 7.05 (br. s., 1H), 7.13-7.33 (m, 1H), 7.38-7.52 (m, 1H), 7.58 (br. s., 1H), 8.26-8.53 (m, 1H); MS obsd. (ESI.sup.+) [(M+H).sup.+]: 303.

Example 2: 9,10-Dimethoxy-6-methyl-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(142) ##STR00018##

Step 1: Preparation of tert-butyl 9,10-dimethoxy-6-methyl-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylate

(143) ##STR00019##

(144) A mixture of tert-butyl 9,10-dimethoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylate (100 mg, 0.28 mmol), K.sub.2CO.sub.3 (77 mg, 0.56 mmol) and CH.sub.3I (174 L, 2.8 mmol) in acetonitrile (10 mL) was heated at 90 C. with stirring for 3 hrs. The mixture was cooled down to rt and diluted with water (10 mL). The resulting mixture was extracted with EtOAc (20 mL) for three times. The combined organic layers were washed with brine, dried over anhydrous Na.sub.2SO.sub.4, and concentrated to give crude tert-butyl 9,10-dimethoxy-6-methyl-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylate (95 mg) which was used in the next step without further purification.

Step 2: Preparation of 9,10-dimethoxy-6-methyl-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(145) ##STR00020##

(146) A mixture of tert-butyl 9,10-dimethoxy-6-methyl-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylate (95 mg, 0.26 mmol) and CF.sub.3COOH (1 mL) in DCM (10 mL) was stirred at rt for 3 hrs. The mixture was concentrated under reduced pressure and the residue was purified by prep-HPLC to give 9,10-dimethoxy-6-methyl-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (10 mg) as a light yellow solid, .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 2.65 (s, 3H) 3.81-3.94 (m, 7H) 4.33-4.43 (m, 2H) 7.00-7.11 (m, 1H) 7.50-7.57 (m, 1H) 7.60-7.66 (m, 1H) 8.57 (s, 1H); MS obsd. (ESI.sup.+) [(M+H).sup.+]: 317.

Example 3: 6-Allyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(147) ##STR00021##

Step 1: Preparation of tert-butyl 6-allyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylate

(148) ##STR00022##

(149) To a solution of tert-butyl 9,10-dimethoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylate (100 mg, 279 mol) in DMF (3 mL) was added allyl bromide (50.6 mg, 419 mol) and potassium carbonate (77.1 mg, 558 mol). The resulting mixture was heated at 60 C. with stirring for 16 hrs, and then filtered. The filtrate was concentrated under reduced pressure to give tert-butyl 6-allyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylate which was used in the next step without further purification.

Step 2: Preparation of 6-allyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(150) ##STR00023##

(151) To a solution of crude tert-butyl 6-allyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylate (100 mg) in DCM (2 mL) was added 2,2,2-trifluoroacetic acid (65 L). The mixture was stirred for 5 hours at rt, and then concentrated under reduced pressure. The residue was purified by prep-HPLC to give 6-allyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (28 mg), .sup.1H NMR (400 MHz, DMSO) ppm 8.46 (s, 1H), 7.64 (s, 1H), 7.55 (s, 1H), 7.06 (s, 1H), 6.04-5.87 (m, 1H), 5.17 (m, 1H), 5.03 (m, 1H), 4.53-4.36 (m, 2H), 3.91 (s, 3H), 3.86 (s, 3H), 3.41 (d, 2H); MS obsd. (ESI.sup.+) [(M+H).sup.+]: 343.

Example 4: 6-Isopropyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(152) ##STR00024##

Step 1: Preparation of tert-butyl 6-isopropyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylate

(153) ##STR00025##

(154) To a solution of tert-butyl 9,10-dimethoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylate (60 mg, 167 mol) in DMF (2 mL) was added potassium carbonate (46.3 mg, 335 mol) and diisopropyl sulfate (61 mg, 335 mol). The mixture was heated at 80 C. with stirring for 14 hours, and then filtered. The filtrate was concentrated under reduced pressure to give crude tert-butyl 6-isopropyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylate, which was used in the next step without further purification.

Step 2: Preparation of 6-isopropyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(155) ##STR00026##

(156) To a solution of crude tert-butyl 6-isopropyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylate (62 mg) in DCM (5 mL) was added TFA (62 L). The mixture was stirred for 3 hours at rt, and then concentrated under reduced pressure. The residue was purified by prep-HPLC to give 6-isopropyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (4 mg), .sup.1H NMR (400 MHz, DMSO) ppm 8.16 (s, 1H), 7.81 (s, 1H), 7.73 (s, 1H), 7.13 (s, 1H), 4.37-4.28 (m, 2H), 3.94 (s, 3H), 3.89 (s, 3H), 2.53-2.51 (m, 1H), 1.39 (d, 6H); MS obsd. (ESI.sup.+) [(M+H).sup.+]: 345.

Example 5: 6-Isobutyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(157) ##STR00027##

Step 1: Preparation of tert-butyl 9,10-dimethoxy-6-(2-methylallyl)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylate

(158) ##STR00028##

(159) To a solution of tert-butyl 9,10-dimethoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylate (100 mg, 279 mol) in DMF (3 mL) was added 3-bromo-2-methylprop-1-ene (75.3 mg, 558 mol) and potassium carbonate (77.1 mg, 558 mol). The mixture was heated at 60 C. with stirring for 16 hours, and then filtered. The filtrate was concentrated under reduced pressure to give crude tert-butyl 9,10-dimethoxy-6-(2-methylallyl)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylate (130 mg) which was used in the next step without further purification.

Step 2: Preparation of 9,10-dimethoxy-6-(2-methylallyl)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(160) ##STR00029##

(161) To a solution of crude tert-butyl 9,10-dimethoxy-6-(2-methylallyl)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylate (130 mg, 268 mol) in DCM (2 mL) was added TFA (99.2 L). The mixture was stirred at rt for 5 hours, and then concentrated under reduced pressure. The residue was purified by column chromatography to give 9,10-dimethoxy-6-(2-methylallyl)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (30 mg).

Step 3: Preparation of 6-isobutyl-9,10-dimethoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(162) ##STR00030##

(163) To a solution of 9,10-dimethoxy-6-(2-methylallyl)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (30 mg, 84.2 mol) in EtOH (3 mL) was added Pd/C (6 mg). The mixture was stirred under an atmosphere of hydrogen at rt for 16 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure to give 6-isobutyl-9,10-dimethoxy-2-oxo-6,7-dihydro-2H-pyrido[2,1-a]phthalazine-3-carboxylic acid (13 mg), .sup.1H NMR (400 MHz, DMSO) ppm 8.40 (s, 1H), 7.62 (s, 1H), 7.55 (s, 1H), 7.09 (s, 1H), 4.42 (m, 2H), 3.90 (s, 3H), 3.86 (s, 3H), 2.55 (d, 2H), 1.69-1.56 (m, 1H), 0.91 (d, 6H); MS obsd. (ESI.sup.+) [(M+H).sup.+]: 359.

Example 6: 6-Isobutyl-10-methoxy-9-(3-methoxypropoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(164) ##STR00031##

Step 1: Preparation of 5-benzyloxy-2-bromo-4-methoxy-benzaldehyde

(165) ##STR00032##

(166) To a solution of 2-bromo-5-hydroxy-4-methoxy-benzaldehyde (25 g, 108 mmol) in MeCN (500 mL) was added (chloromethyl)benzene (13.7 g, 108 mmol) and potassium carbonate (29.9 g, 216 mmol). The mixture was heated at 80 C. with stirring for 15 hours. After being cooled to rt, the mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was partitioned between brine and DCM. The organic layer was dried over anhydrous Na.sub.2SO.sub.4 and concentrated under reduced pressure to give 5-(benzyloxy)-2-bromo-4-methoxy-benzaldehyde (34.2 g, 96.9 mmol) which was used in the next step without further purification.

Step 2: Preparation of (5-benzyloxy-2-bromo-4-methoxy-phenyl)methanol

(167) ##STR00033##

(168) To a solution of 5-(benzyloxy)-2-bromo-4-methoxy-benzaldehyde (35 g, 106 mmol) in MeOH (600 mL) was added NaBH.sub.4 (4 g, 106 mmol) portion-wise at 0 C. The reaction mixture was allowed to warm to rt, stirred at rt for three hours, and then quenched with water. The mixture was concentrated under reduced pressure. The residue was partitioned between brine and DCM. The organic layer was dried over anhydrous Na.sub.2SO.sub.4 and concentrated under reduced pressure to give (5-(benzyloxy)-2-bromo-4-methoxy-phenyl)methanol (32.5 g, 100 mmol), which was used in the next step without further purification.

Step 3: Preparation of 1-benzyloxy-4-bromo-5-(chloromethyl)-2-methoxy-benzene

(169) ##STR00034##

(170) To a solution of (5-(benzyloxy)-2-bromo-4-methoxy-phenyl)methanol (5 g, 14.7 mmol) in DCM (50 mL) was added SOCl.sub.2 (1.61 mL, 22 mmol) drop-wise at rt. The mixture was stirred at rt for two hours, and then concentrated under reduced pressure to give 1-(benzyloxy)-4-bromo-5-(chloromethyl)-2-methoxy-benzene (5 g, 13.8 mmol, 93.6% yield), which was used in the next step without further purification.

Step 4: Preparation of tert-butyl 1-[(5-benzyloxy-2-bromo-4-methoxy-phenyl)methyl-tert-butoxycarbonyl-amino]-4-oxo-pyridine-3-carboxylate

(171) ##STR00035##

(172) To a solution of 1-(benzyloxy)-4-bromo-5-(chloromethyl)-2-methoxy-benzene (5.0 g, 14.6 mmol) in MeCN (100 mL) was added potassium carbonate (4.05 g, 29.3 mmol) and tert-butyl 1-((tert-butoxycarbonyl)amino)-4-oxo-1,4-dihydropyridine-3-carboxylate (14 g, 29.3 mmol). The mixture was heated at 80 C. with stirring for 5 hours. After being cooled to rt, the mixture was filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to give tert-butyl 1-[(5-benzyloxy-2-bromo-4-methoxy-phenyl)methyl-tert-butoxycarbonyl-amino]-4-oxo-pyridine-3-carboxylate (5.3 g, 7.75 mmol).

Step 5: Preparation of tert-butyl 9-benzyloxy-10-methoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylate and di-tert-butyl 9-benzyloxy-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3,6-dicarboxylate

(173) ##STR00036##

(174) To a 15 mL microwave vial was added tert-butyl 1-[(5-benzyloxy-2-bromo-4-methoxy-phenyl)methyl-tert-butoxycarbonyl-amino]-4-oxo-pyridine-3-carboxylate (700 mg, 1.14 mmol), palladium (II) bromide (90.8 mg, 341 mol), potassium acetate (223 mg, 2.27 mmol) and DMF (5 mL). The vial was capped and heated under microwave at 100 C. for 5 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure to give a mixture of tert-butyl 9-benzyloxy-10-methoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylate and di-tert-butyl 9-benzyloxy-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3,6-dicarboxylate, which was used in the next step without further purification.

Step 6: Preparation of 9-benzyloxy-10-methoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylic acid

(175) ##STR00037##

(176) To a solution of the crude mixture of tert-butyl 9-benzyloxy-10-methoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylate and di-tert-butyl 9-benzyloxy-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3,6-dicarboxylate (4.0 g) in DCM (20 mL) was added TFA (20 mL). The mixture was stirred at rt for 15 hours, and then concentrated under reduced pressure. The residue was treated with a mixture solvent of petroleum ether/ethyl acetate (10:1, V/V). The mixture was filtered and the filter cake was dried under reduced pressure to give 9-benzyloxy-10-methoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylic acid (2.0 g).

Step 7: Preparation of methyl 9-benzyloxy-10-methoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylate

(177) ##STR00038##

(178) To a suspension of 9-benzyloxy-10-methoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylic acid (0.70 g, 1.85 mmol) in MeOH (20 mL) was added catalytic amount of concentrated sulfuric acid (50 L). The mixture was heated at 80 C. for 20 hours. The mixture was concentrated under reduced pressure, and the residue was neutralized with saturated aqueous NaHCO.sub.3. The resulting mixture was extracted with DCM. The organic layer was dried over anhydrous Na.sub.2SO.sub.4 and concentrated under reduced pressure to give methyl 9-benzyloxy-10-methoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylate (703 mg, 1.79 mmol).

Step 8: Preparation of 9-benzyloxy-10-methoxy-6-(2-methylallyl)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(179) ##STR00039##

(180) To a solution of methyl 9-benzyloxy-10-methoxy-2-oxo-6,7-dihydropyrido[2,1-a]phthalazine-3-carboxylate (0.20 g, 510 mol) in DMF (20 mL) was added sodium hydride (61.2 mg, 1.53 mmol) at 0 C. The mixture was stirred for 30 minutes, and then 3-bromo-2-methylprop-1-ene (275 mg, 2.04 mmol) was added. The resulting mixture was allowed to warm to rt and stirred at rt for 40 hours. To the mixture was added water and sodium hydroxide (40.8 mg, 1.02 mmol). The resulting mixture was stirred for additional two hours, and then acidified with 6 M hydrochloric acid, and extracted with DCM. The organic layer was dried over anhydrous Na.sub.2SO.sub.4 and concentrated under reduced pressure to give crude 9-benzyloxy-10-methoxy-6-(2-methylallyl)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (241 mg), which was used in the next step without further purification.

Step 9: Preparation of 9-hydroxy-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(181) ##STR00040##

(182) To a solution of crude 9-benzyloxy-10-methoxy-6-(2-methylallyl)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (241 mg) in EtOH (15 mL) was added Pd/C (50 mg). The mixture was stirred under hydrogen atmosphere at rt for 16 hours. Then the mixture was filtered and the filtrate was concentrated under reduced pressure to give crude 9-hydroxy-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (123 mg), which was used in the next step without further purification.

Step 10: Preparation 6-isobutyl-10-methoxy-9-(3-methoxypropoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(183) ##STR00041##

(184) To a solution of crude 9-hydroxy-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (60 mg) in DMF (3 mL) was added potassium carbonate (74.2 mg, 537 mol) and 1-bromo-3-methoxypropane (61.6 mg, 402 mol). The mixture was heated at 100 C. with stirring for three hours, and then cooled to rt. To the mixture was added water (3 mL) and sodium hydroxide (10.7 mg, 268 mol). The resulting mixture was stirred at rt for two hours, then acidified with 6 M hydrochloric acid and extracted with DCM. The organic layer was concentrated under reduced pressure, and the residue was purified by prep-HPLC to give 6-isobutyl-10-methoxy-9-(3-methoxypropoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (17 mg), .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 8.40 (s, 1H), 7.62 (s, 1H), 7.55 (s, 1H), 7.10 (s, 1H), 4.46-4.36 (m, 2H), 4.11 (t, 2H), 3.90 (s, 3H), 3.48 (t, 2H), 3.26 (s, 3H), 2.55 (d, 2H), 2.05-1.95 (m, 2H), 1.69-1.57 (m, 1H), 0.91 (d, 6H); MS obsd. (ESI.sup.+) [(M+H).sup.+]: 417.

Example 7: 6-Isobutyl-10-methoxy-2-oxo-9-(2,2,2-trifluoroethoxy)-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(185) ##STR00042##

(186) To a solution of crude 9-hydroxy-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (60 mg) in DMF (3 mL) was added potassium carbonate (74.2 mg, 537 mol) and 1,1,1-trifluoro-2-iodoethane (169 mg, 805 mol). The mixture was heated at 100 C. with stirring for 20 hours. After being cooled to rt, the reaction mixture was partitioned between 1 M hydrochloric acid and DCM. The organic layer was concentrated under reduced pressure, and the residue was purified by prep-HPLC to give 6-isobutyl-10-methoxy-2-oxo-9-(2,2,2-trifluoroethoxy)-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (7 mg), .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 8.42 (s, 1H), 7.72 (s, 1H), 7.62 (s, 1H), 7.22 (s, 1H), 4.90-4.78 (m, 2H), 4.48-4.35 (m, 2H), 3.94 (s, 3H), 2.54 (d, 2H), 1.66-1.56 (m, 1H), 0.91 (d, 6H); MS obsd. (ESI.sup.+) [(M+H)+]: 427.

Example 8: 6-Isobutyl-10-methoxy-2-oxo-9-(tetrahydropyran-4-ylmethoxy)-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(187) ##STR00043##

(188) To a solution of crude 9-hydroxy-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (60 mg) in DMF (3 mL) was added potassium carbonate (74.2 mg, 537 mol) and 4-(iodomethyl)tetrahydro-2H-pyran (91 mg, 402 mol). The mixture was heated at 100 C. with stirring for 3 hrs, and then cooled to rt. To the above mixture was added water (3 mL) and sodium hydroxide (10.7 mg, 268 mol). The resulting mixture was stirred at rt for 2 hrs, and then acidified with 6 M hydrochloric acid. The mixture was partitioned between brine and DCM. The organic layer was separated and concentrated under reduced pressure. The residue was purified by prep-HPLC to give 6-isobutyl-10-methoxy-2-oxo-9-(tetrahydropyran-4-ylmethoxy)-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (5 mg), .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 8.60 (s, 1H), 7.20 (s, 1H), 7.04 (s, 1H), 6.72 (s, 1H), 4.36-4.16 (m, 2H), 4.08-4.01 (m, 2H), 3.94 (s, 3H), 3.92 (d, 2H), 3.52-3.43 (m, 2H), 2.53 (d, 2H), 2.24-2.19 (m, 1H), 2.05-1.99 (m, 1H), 1.69-1.60 (m, 2H), 1.54-1.42 (m, 2H), 0.96 (d, 6H). MS obsd. (ESI.sup.+) [(M+H).sup.+]: 443.

Example 9: 6-Isobutyl-10-methoxy-9-(3-methylsulfonylpropoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(189) ##STR00044##

(190) To a solution of crude 9-hydroxy-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (60 mg) in DMF (3 mL) was added potassium carbonate (74.2 mg, 537 mol) and 3-(methylsulfonyl)propyl 4-methylbenzenesulfonate (118 mg, 402 mol). The mixture was heated at 100 C. with stirring for 3 hrs, and then cooled to rt. To the above mixture was added water (3 mL) and sodium hydroxide (10.7 mg, 268 mol). The resulting mixture was stirred at rt for 2 hrs, and then acidified with 6 M hydrochloric acid. The mixture was partitioned between brine and DCM. The organic layer was separated and concentrated under reduced pressure. The residue was purified by prep-HPLC to give 6-isobutyl-10-methoxy-9-(3-methylsulfonylpropoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (7.8 mg), .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 8.40 (s, 1H), 7.64 (s, 1H), 7.55 (s, 1H), 7.09 (s, 1H), 4.44-4.38 (m, 2H), 4.22-4.15 (m, 2H), 3.91 (s, 3H), 3.27-3.22 (m, 2H), 3.04 (s, 3H), 2.54 (d, 2H), 2.23-2.16 (m, 2H), 1.66-1.56 (m, 1H), 0.91 (d, 6H); MS obsd. (ESI.sup.+) [(M+H).sup.+]: 465.

Example 10: 6-Isobutyl-10-methoxy-9-(2-morpholinoethoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(191) ##STR00045##

(192) To a solution of crude 9-hydroxy-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (57 mg) in DMF (3 mL) was added potassium carbonate (88.1 mg, 637 mol) and 4-(2-bromoethyl)morpholine hydrobromide (87.6 mg, 319 mol) The mixture was heated at 100 C. with stirring for 3 hrs, and then cooled to rt. To the above mixture was added water (3 mL) and sodium hydroxide (10.2 mg, 255 mol). The resulting mixture was stirred at rt for 2 hrs, and then acidified with 6 M hydrochloric acid. The mixture was partitioned between brine and DCM. The organic layer was separated and concentrated under reduced pressure. The residue was purified by prep-HPLC to give 6-isobutyl-10-methoxy-9-(2-morpholinoethoxy)-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (10 mg), .sup.1H NMR (400 MHz, CDCl.sub.3) ppm 8.60 (s, 1H), 7.21 (s, 1H), 7.05 (s, 1H), 6.75 (s, 1H), 4.30-4.23 (m, 4H), 3.95 (s, 3H), 3.76-3.73 (m, 4H), 2.93-2.89 (m, 2H), 2.65-2.62 (m, 4H), 2.53 (d, 2H), 1.68-1.61 (m, 1H), 0.97 (d, 6H); MS obsd. (ESI.sup.+) [(M+H).sup.+]: 458.

Example 11: 9-(3-Hydroxy-2,2-dimethyl-propoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(193) ##STR00046##

(194) To a solution of crude 9-hydroxy-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (57 mg, 127 mol) in DMF (3 mL) was added potassium carbonate (88.1 mg, 637 mol) and 3-bromo-2,2-dimethylpropan-1-ol (85.2 mg, 510 mol). The mixture was heated at 100 C. with stirring for 3 hrs, and then cooled to rt. To the above mixture was added water (3 mL) and sodium hydroxide (10.2 mg, 255 mol). The resulting mixture was stirred at rt for 2 hrs, and then acidified with 6 M hydrochloric acid. The mixture was partitioned between brine and DCM. The organic layer was separated and concentrated under reduced pressure. The residue was purified by prep-HPLC to give 9-(3-hydroxy-2,2-dimethyl-propoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (7 mg), .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 8.39 (s, 1H), 7.61 (s, 1H), 7.52 (s, 1H), 7.09 (s, 1H), 4.67-4.61 (m, 1H), 4.42-4.37 (m, 2H), 3.90 (s, 3H), 3.78 (s, 2H), 3.30-3.28 (m, 2H), 2.54 (d, 2H), 1.65-1.58 (m, 1H), 0.95 (s, 6H), 0.90 (d, 6H); MS obsd. (ESI.sup.+) [(M+H).sup.+]: 431.

Example 12: 9-(4-Carboxybutoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(195) ##STR00047##

(196) To a solution of crude 9-hydroxy-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (57 mg, 127 mol) in DMF (3 mL) was added potassium carbonate (88.1 mg, 637 mol) and methyl 5-bromopentanoate (74.6 mg, 382 mol). The mixture was heated at 100 C. with stirring for 3 hrs, and then cooled to rt. To the above mixture was added water (3 mL) and sodium hydroxide (20.4 mg, 510 mol). The resulting mixture was stirred at rt for 2 hrs, and then acidified with 6 M hydrochloric acid. The mixture was partitioned between brine and DCM. The organic layer was separated and concentrated under reduced pressure. The residue was purified by prep-HPLC to give 9-(4-carboxybutoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (7 mg), .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 8.39 (s, 1H), 7.61 (s, 1H), 7.53 (s, 1H), 7.08 (s, 1H), 4.44-4.36 (m, 2H), 4.09-4.03 (m, 2H), 3.90 (s, 3H), 2.53 (d, 2H), 2.33-2.26 (m, 2H), 1.81-1.73 (m, 2H), 1.69-1.59 (m, 3H), 0.91 (d, 6H). MS obsd. (ESI.sup.+) [(M+H).sup.+]: 445.

Example 13: 9-(3-Cyanopropoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(197) ##STR00048##

(198) To a solution of crude 9-hydroxy-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (57 mg, 127 mol) in DMF (3 mL) was added potassium carbonate (88.1 mg, 637 mol) and 4-bromobutanenitrile (56.6 mg, 382 mol). The mixture was heated at 100 C. with stirring for 3 hrs, and then cooled to rt. To the above mixture was added water (3 mL) and sodium hydroxide (10.2 mg, 255 mol). The resulting mixture was stirred at rt for 2 hrs, and then acidified with 6 M hydrochloric acid. The mixture was partitioned between brine and DCM. The organic layer was separated and concentrated under reduced pressure. The residue was purified by prep-HPLC to give 9-(3-cyanopropoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (10 mg), .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 8.40 (s, 1H), 7.63 (s, 1H), 7.55 (s, 1H), 7.12 (s, 1H), 4.45-4.36 (m, 2H), 4.16-4.09 (m, 2H), 3.90 (s, 3H), 2.70-2.62 (m, 2H), 2.54 (d, 2H), 2.09-2.05 (m, 2H), 1.66-1.57 (m, 1H), 0.91 (d, 6H). MS obsd. (ESI.sup.+) [(M+H).sup.+]: 412.

Example 14: 9-[5-(Tert-butoxycarbonylamino)pentoxy]-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid

(199) ##STR00049##

(200) To a solution of crude 9-hydroxy-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (57 mg, 127 mol) in DMF (3 mL) was added potassium carbonate (88.1 mg, 637 mol) and 5-((tert-butoxycarbonyl)amino)pentyl 4-methylbenzenesulfonate (137 mg, 382 mol). The mixture was heated at 100 C. with stirring for 3 hrs, and then cooled to rt. To the above mixture was added water (3 mL) and sodium hydroxide (10.2 mg, 255 mol). The resulting mixture was stirred at rt for 2 hrs, and then acidified with 6 M hydrochloric acid. The mixture was partitioned between brine and DCM. The organic layer was separated and concentrated under reduced pressure. The residue was purified by prep-HPLC to give 9-[5-(tert-butoxycarbonylamino)pentoxy]-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (13 mg), .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 8.39 (s, 1H), 7.61 (s, 1H), 7.53 (s, 1H), 7.08 (s, 1H), 6.83-6.77 (m, 1H), 4.43-4.36 (m, 2H), 4.07-4.00 (m, 2H), 3.89 (s, 3H), 2.97-2.89 (m, 2H), 2.54 (d, 2H), 1.79-1.71 (m, 2H), 1.66-1.56 (m, 1H), 1.46-1.34 (m, 13H), 0.90 (d, 6H). MS obsd. (ESI.sup.+) [(M+H).sup.+]: 530.

Example 15: 9-(5-Aminopentoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid hydrochloride

(201) ##STR00050##

(202) To a solution of 9-[5-(tert-butoxycarbonylamino)pentoxy]-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid (8 mg, 15.1 mol) in MeCN (0.5 mL) was added hydrochloric acid (5.51 L, 181 mol). The mixture was stirred at rt for 2 hrs, and then concentrated under reduced pressure to 9-(5-aminopentoxy)-6-isobutyl-10-methoxy-2-oxo-7H-pyrido[2,1-a]phthalazine-3-carboxylic acid hydrochloride (7 mg), .sup.1H NMR (400 MHz, DMSO-d.sub.6) ppm 8.40 (s, 1H), 7.90-7.79 (m, 3H), 7.61 (s, 1H), 7.54 (s, 1H), 7.09 (s, 1H), 4.45-4.36 (m, 2H), 4.09-4.02 (m, 2H), 3.90 (s, 3H), 2.85-2.76 (m, 2H), 2.56-2.52 (m, 2H), 1.83-1.77 (m, 2H), 1.67-1.58 (m, 3H), 1.52-1.42 (m, 2H), 0.91 (d, 6H); MS obsd. (ESI.sup.+) [(M+H).sup.+]: 430.

Biological Examples

Example 16 Materials and Methods

(203) HBV Cell Line

(204) HepG2.2.15 cells (Acs et al. Proc Natl Acad Sci USA, 84, (1987), 4641-4), a constitutively HBV-expressing cell line were cultured in DMEM+Glutamax-I medium (Invitrogen, Carlsbad, Calif., USA), supplemented with 10% fetal bovine serum (Invitrogen) and G418 (Invitrogen) at a final concentration of 200 mg/L and maintained in 5% CO.sub.2 at 37 C.

(205) HBsAg Assay

(206) HepG2.2.15 cells were seeded in duplicate into white, 96-well plates at 1.510.sup.4 cells/well. The cells were treated with a three-fold serial dilution series of the compounds in DMSO. The final DMSO concentration in all wells was 1% and DMSO was used as no drug control.

(207) The HBsAg chemiluminescence immunoassay (CLIA) kit (Autobio Diagnostics Co., Zhengzhou, China, Catalog number: CL0310-2) was used to measure the levels of secreted HBV antigens semi-quantitatively. For the detection 50 L/well culture supernatant was used and HBsAg was quantified using HBsAg chemiluminescence immunoassay (CLIA) kit (Autobio Diagnostics Co., Zhengzhou, China, Catalog number: CL0310-2), 50 L of the supernatant was transferred to the CLIA assay plate and 50 L of enzyme conjugate reagent was added into each well. The plates were sealed and gently agitated for 1 hour at room temperature. The supernatant-enzyme-mixture was discarded and wells were washed 6 times with 300 L of PBS. The residual liquid was removed by plating the CLIA plate right side down on absorbent tissue paper. 25 L of substrates A and B were added to each well. Luminance was measured using a luminometer (Mithras LB 940 Multimode Microplate Reader) after 10 minutes incubation. Dose-response curves were generated and the IC.sub.50 value was extrapolated by using the E-WorkBook Suite (ID Business Solutions Ltd., Guildford, UK). The IC.sub.50 was defined as the compound concentration (or conditioned media log dilution) at which HBsAg secretion was reduced by 50% compared to the no drug control.

(208) The compounds according to formula I were tested for their capacity to inhibit HBsAg as described herein. The Examples were tested in the above assay and found to have IC.sub.50 below 50 M. Particular compounds of formula I were found to have IC.sub.50 below 1.0 M. Results of HBsAg assay are given in Table 1.

(209) TABLE-US-00003 TABLE 1 List of Activity data Example No. IC.sub.50 (M) 1 18.707 2 2.534 3 0.74 4 26.068 5 0.248 6 0.053 7 0.172 8 0.044 9 0.624 10 0.225 11 0.039 12 9.428 13 0.11 14 0.047 15 0.5