AU684777B2 - Substituted methylenedioxy(3',4':6,7)indolizino-(1,2-b)quinolinones - Google Patents

Description

WO 94/25465 PCT/US94/04866 SUBSTITUTED METHYLENEDIOXY[3',4':6,7]INDOLIZINO- [1,2-b]QUINOLINONES This patent application is a continuation-in-part of U.S. Serial No.

08/057,133, filed on May 3, 1993 by Pendrak et al.

SCOPE OF THE INVENTION This invention relates to antiviral compounds, pharmaceutical compositions thereof, and a method of treating viral infections. More specifically, this invention relates to certain indolizino[1,2-b]-quinolinyl derivatives which have antiviral activity.

BACKGROUND OF THE INVENTION Certain 1H-pyrano[3',4':6,7]indolizino[1,2-b)quinolinones are known to have cytotoxic and antiviral activity. Camptothecin is an example of one such compound.

It is a water-insoluble, cytotoxic alkaloid produced by Camptotheca acuminata trees indigenous to China and Nothapodytes foda trees indigenous to India.

Camptothecin and its close congeners are known to inhibit eukaryotic topoisomerase I. The cytotoxic and antitumor activity of camptothecin and its close congeners is due to this inhibition of eukaryotic topoisomerase I (Cancer Res. 1988, 48, 1722; Molec. Pharmacol. 1988, 34, 755). Compounds that are related in structure to camptothecin but do not inhibit eukaryotic topoisomerase I are not cytotoxic to mammalian cells and have no antitumor activity Med. Chem. 1988, 32, 715; Cancer Res. 1989, 49, 1465; Cancer Res. 1989, 49, 4358).

Camptothecin has been shown to have an effect on viruses by a number of investigators in laboratory settings. Although camptothecin has demonstrated antiviral activity in in vitro tissue culture systems, camptothecin and its close analogs that have a hydroxylactone moiety cannot be considered as useful in vivo antiviral agents because they inhibit mammalian topoisomerase I, inhibit host cell DNA repication, and are cytotoxic to mammalian cells. Furthermore, camptothecin is not expected to be attractive for drug development as an antiviral agent because of unacceptable dose-limiting toxicity, unpredictable toxicity, poor aqueous solubility, and/or unacceptable shelf life stability.

There is a need for new antiviral agents. Some substituted 1Hpyrano[3',4':6,7]indolizino-[1,2-b]quinolinones that lack the E-ring ct-hydroxy lactone moiety of camptothecin have been shown to be non-cytotoxic to mammalian cells and to lack antitumor activity (Ann. Rev. Pharmcol. Toxicol. 1977, 17, 117; J.

Med. Chem. 1989, 32, 715). This is because these compounds do not contain the I I- ll~l~ rpl WO 94/25465 WO 9425465PCT/US94/04866 essential structural features required to inhibit eukaryotic topoisomerase 1. But it has been found that certain 7-ethyl-7-hydroxy-7,8,l 1,13-tetrahydro-IOdioxolo[4,5g]pyrano[3',4':6,7lindolizino[ 1,2-b]quinolinone-8, 1-diones (hereinafter "methylenedioxyindolizino[1,2-b]quinolinones") lacking the hydroxylactone moiety do have antiviral activity without the undesirable features of caxnptothecin. As such they are useful for treating viral infections.

SUMMARY OF JE TNVEnIO .0 In a first aspect, the present invention provides a method for treating viral infections, which method comprises administering to an infected host in need thereof an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt thereof, alone or in combination with a carrier, diluent or excipient.

wherein: R is -OH, and OR I; RI is -COR 4 or -P(O)(OH)R 5 wherein:

R

3 is -H or lower allI;

R

4 is -CR 3

R

6

R

7

-(CH

2 )nCH2R 7 (where -(CH2)nCH2COOH (where WO 94/25465 PCT/US94/04866

-NH(CH

2 )nCH2R 7 (where n and

-NH(CH

2 )nCH 2 COOH (where n 0-3);

R

5 is OH or CH 2

NH

2

R

6 is H or the side chain of any naturally occuring a-amino acid; -ND N N

R

7 is -NR 9

R

10 where X is any pharmaceutically acceptable anion;

R

8 is lower alkyl;

R

9 and R 10 are independently selected from the group consisting of -H, -C1-6 alkyl, and R 9 and R10 taken together to form a 5-7 membered saturated heterocyclic ring containing the nitrogen on which R 9 and R 10 are substituted; and

R

11 is -CH 2

R

12 wherein: -N 0, -N NCH,, -OH

R

12 is -N(CH 3 2 2 In another aspect, this invention relates to novel compounds of Formula I, and pharmaceutically acceptable salts thereof.

In yet another aspect, this invention relates to a composition comprising a novel compound of Formula I in combination with an acceptable carrier, excipient or diluent, particularly a pharmaceutically acceptable carrier, excipient or diluent.

WO 94/25465 PCT/US94/04866 DETAILED DESCRIPTION OF THE INVENTION Definitions The terms below, defined as follows, are used in describing the present invention throughout this application.

"Aliphatic" is intended to include saturated and unsaturated radicals. This includes normal and branched chains, saturated or mono or poly unsaturated chains where both double and triple bonds may be present in any combination. The phrases "lower alkyl" and "C 1 -6 alkyl" refer to and mean an alkyl group of 1 to 6 carbon atoms in any isomeric form, but particularly the normal or linear form. "Lower alkoxy" means the group lower alkyl-O-. "Halo" means fluoro, chloro, bromo or iodo. "Acyl" means the radical having a terminal carbonyl carbon.

The phrase "5-7 membered saturated heterocyclic ring containing the nitrogen" is intended to include saturated rings such as piperidine, pyrrolidine, morpholine, piperazine, and N-alkyl piperazine.

Salts of any sort may be made from these compounds, provided there is an acidic group present or a sufficiently basic nitrogen. Particularly preferred are the pharmaceutically acceptable salts of the instant compounds. These latter salts are those which are acceptable in their application to a pharmaceutical use. By that it is meant that the salt will retain the biological activity of the parent compound and the salt will not have untoward or deleterious effects in its application and use in treating diseases.

Pharmaceutically acceptable salts are prepared in a standard manner. The parent compound in a suitable solvent is reacted with an excess of an organic or inorganic acid, in the case of acid addition salts of a base moiety, or an excess of organic or inorganic base in the case where there is an acid group. Representative acids are hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, acetic acid, maleic acid, succinic acid or methanesulfonic acid. Cationic salts are readily prepared from metal bases such as sodium, potassium, calcium, magnesium, zinc, copper or the like and ammonia. Organic bases include the mono or disubstituted amines, ethylenediamine, piperazine, amino acids, caffeine, and the like.

I

WO 94/25465 PCT/US94/04866 Here and throughout this application, the ring system of the compounds of the present invention are numbered according to Formula II.

1 D 2 11 2 N 9

R

II

If a chiral center or another form of an isomeric center is created by some cr.lbination of substituents, in a compound of the present invention, all forms of such isomer(s) are intended to be covered herein. Inventive compounds containing a chiral center may be used as a racemic mixture or the mixture may be separated and an individual enantiomer may be used alone.

The present invention provides a method for the treatment of viral infections caused by certain DNA viruses comprising administering to an infected animal, preferably a mammal, most preferably a human, in need thereof an effective amount of a compound of Formula I as described hereinabove, or a pharmaceutically acceptable salt thereof, alone or in combination with a carrier, excipient or diluent The present invention also provides compounds, and pharmaceutically acceptable salts thereof, which exhibit antiviral activity, said compounds having the structure represented by Formula I hereinabove.

More specifically, these compounds and the present method are especially useful in treating the following pathogens in humans: Herpes Simplex virus types 1 and 2 (HSV-1 and HSV-2); Cytomegalovirus (CMV); Varicella Zoster Virus (VZV); No unacceptable toxicological effects are expected when compounds of the present invention are administered in accordance with the present invention.

A preferred method for treating viral infections according to the present invention uses the following compounds of Formula I: 7-Acetyl-8-methyldioxolo[4,5-g]indolizino[1,2-b]quinolin-9(1 H)-one; (±)-7-(1-Hydroxyethyl)-8-methyldioxolo[4,5-g]indolizino[1,2-b]quinolin- 9(11H)-one; (±)-7-[1(Aminoacetyl)oxy]ethyl]-8-methyldioxolo[4,5-g]indolizino[1,2b]quinolin-9(1 1H)-one hydrotrifluoroacetate;

I

WO 94/25465 PCT/US9404866 7-Acetyl-12-dimethylaminomethyl-8-methyldioxolo indolizino [1,2-bquinolin-9(1 1H)-one; and 7-Acetyl-12*,hydroxymethyl-8-methyldioxolo indolizino [1,2-b] quinolin-9(1 1H)-one.

Preferred compounds of the present invention include: 7-Acetyl-8-methyldioxolo[4,5-g]indoiizino[ 1,2-b]quinolin-9(1 1H)-one; (±)-7-(1-Hydroxyethyl)-8-methyldioxolo[4,5-gjindolizino[l,2-b)quinolin- 9(11H)-one; 1 (Aminoacetyl)oxy]ethyl]-8-methyldioxolo[4,5-g]indolizino[ 1,2b]quinolin-9(11H)-one hydrotrifluoroacetate; 7-Acetyl-12-dimethylaminomethyl-8-methyldioxolo indolizino [1,2-bquinolin-9(11H)-one; and 7-Acetyl-12-hydroxymethyl-8-methyldioxolo indolizino [1,2-b] quinolin-9( 1H)-one.

The compounds of the present invention can be prepared by the following methods.

There are several methods for preparing these compounds. One generic process comprises preparing a 1-keto indolizine adduct and then condensing this fragment with the appropriate substituted arninobenzaldehyde or aminoactophenone.

Starting materials are commercially available or can be made by published methods.

The reaction sequences are illustrated by Schemes I-IM. Commerically available 2-pyrrolidone 1 can be alkylated with dimethyl sulfate to give ether 2 which can be condensed with acetonedicarboxylate to give indolizine 3. Methylation of 3 with methyl iodide at ambient temperature in the inert solvent produced 4, which can be hydrolyzed witi. an aqueous base to give 5 followed by decarboxylation to produce 6.

Triflation of 6 with N-phenyltriflimide in DMF in the presence of base, like triethylamine, can give 7, which can be reacted via Heck reaction with nbutylvinylether in the presence of palladium catalyst and base to give 8. Hydrolysis of 8 with acid, like 3N HC1, can give 9 which can be protected as a ketal using ethyleneglycol and hydrochloride gas to give 10. Compound 10 can be oxidized using Davis reagent and base to give alcohol 11 which can be further oxidized with pyridiniumchlorochromate in methylene chloride to give ketone-ketal 12. Friedlander condensation of compound 12 with aminopiperinal 13 in the presence of p-tolunesulfonic acid in toluene and subsequent hydrolysis with acid, like 3N HC1, can give the title compound 14 (Scheme II). Aminoacetophenone 20 can be prepared in four steps by first nitrating methylenedioxyacetophenone 15 with nitric acid to give compouond 16 which can be brominated with bromine in dioxane to produce compound 17. Reaction of 17 with hydrazine 18 in acetonitrile can give compound -e WO 94/25465 PCT/US94/04866 19 which can be reduced with nickel boride to give amine 20. Condensation of compound 12 and 20 as described previously can give the title compound 21 (Scheme II). Reaction of 17 with with sodium acetate in dimethylformamide produced compound 22 (Scheme II). Reduction of 22 with nickel boride can give compound 23 which can be condensed with compound 12 and hydrolysed as described previously to produce the title compound 24.

I dl WO 94/25465 PTU9146 PCT/US94/04866

SCHEMEI

H

I

Me 2

SO

4 45 0

C

0

EO

2 C CO 2 !Et N 00H 3 Et 3 N 0'

ICO

2 Et Mel /NaH N- THF, r 1 :2taqLiOH MeOH/TH F IC0 2

H

trichlorophenol S 220 0

C

Tf 2 NPh Et 3

N

DMF, rt C OH OH

OSO

2

CF

3 n-Butylvinylether Pd(OAC) 2 dppp N EtN/ DMF 0

PCC

LIDA, -7800 HOI, rt

THF

1

CH

2

CI

2 WO 94/25465 PTU9/46 PCT/US94/04866 SCHEME II 0 0

CHO

0

NH

2 13 PTSA toluene 0 2. 3N HCI /Ac0H .~HN0 3 0 0-40' n Br 2 dioxane

_NZ

CH

3 CN C Ni 2

B

MeOH /HCl 0 0

N

0

NH

2 N0 2 1.

L

12 PTSA toluene 2. 3N HCII/AcOH WO 94125465 WO 941546S CTIUS94104866 SCHEME III Br NaQAc DMF, 67'C 0 0 N0 2 MeOH /HCI N0 2 12 PTSA toluene 2, 3N HCI i AcOH

-NH-

2 WO 94/25465 ICT/VS9A4/4866 The assay used to test the compounds of the present invention for antiviral activity is well-known. A generalized description of the assay follows.

Well plates are seeded with the appropriate cells at a concentration of cells per well suspended in 0.5 mL of Earle's Minimum Essential Medium (EMEM) contining 10% fetal bovine serum (FBS) and antibiotic and antimycotic solution.

After the cells are 80-90% confluent (24 hours), old medium is removea and washed with Hank's buffered saline solution (HBSS). Cells are then infected for 1 hour at 37 0 C with 100-200 plaque forming units per well of a herpes simplex virus suspended in 250 mL HBSS. Following adsorption, the following are acded: A) 250 mL/well 2 x EMEM containing Human IgG (Sigma Chemical Co., St.

Louis, Mo.) (ca. 0.1 mg/mL); B) 250 mLlwell EMEM containing 10% FBS and antibiotic/antimycotic solution; and C) 250 mL/well HBSS containing appropriately diluted compound.

After 24-48 hours (best time determined by observation of plaques under a microscope), old medium is aspirated off. Each well is stained with a selected stain solution crystal violet in MeOH:H20 7:3) and then rinsed with water, air dried, and the plaques are counted. Compound effectiveness is evaluated in terms of percent plaque reduction as compared to untreated, infected controls.

This assay can be used to test compound activity against many other viruses besides herpes simplex by simply modifying the cell type used in the first step to match the virus being tested, and otherwise following the procedure outlined above.

Other cell types which can be used in this assay include mouse mammary tumor cells, human lung fibroblasts, sheep chorioplexus cells, and green monkey kidney cells.

Alternatively, other assays can be used to determine the antiviral activity of the present compounds. Such assays include the following types: cell count, clonogenic, cytopathic effect, dish-colony formation, microtiter-growth inhibition, thymidine incorporation and yield reduction. Each of these assays is well-known and is available either from the literature or from a commercial testing lab.

The present invention provides pharmaceutical compositions prepared from the compounds of Formula I. These compositions have both a human and veterinary utility, and comprise an excipient or carrier which is acceptable for the intended pharmaceutical end use and at least one inventive compound. For example, if a veterinary use is intended, the carrier may be a liquid, or spray, or may be formulated in a solid, non-degradeable or degradeable form for insertion in the rumen. Selected excipients and carriers may be employed to prepare compositions acceptable or adaptable for humans use.

-11- I, I WO 94/25465 PCT/US94/04866 An effective amount of the pharmaceutical compositions of the present invention may be contained in one embodiment, such as in a single pill, capsule, or pre-measured intravenous dose or pre-filled syringe for injection. Alternatively, as is frequently the case, the composition will be prepared in individual dose forms where one unit, such as a pill, will contain a sub-optimal dose out the user will be instructed to take two or more unit doses per treatment. When the composition is presented as a cream, it will contain a discrete amount of drug and the user will apply some amount of the cream one or more times until the disease is in remission or has been effectively treated. Concentrates for later dilution by the end user may also be prepared, for instance for intravenous (IV) formulations and multi-dose injectable formulations.

Carriers or diluents contemplated for use in these compositions are generally known in the pharmaceutical formulary arts. Reference to useful materials can be found in well known compilations such as Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa.

The nature of the composition and the pharmaceutical carrier or diluent will, of course, depend upon the intended route of administration, for example whether by intravenous and intramuscular injection, parenterally, topically, orally, or by inhalation.

For parenteral administration the pharmaceutical composition will be in the form of a sterile injectable liquid such as an ampule or an aqueous or nonaqueous liquid suspension.

For topical administration the pharmaceutical composition will be in the form of a cream, ointment, linimnt, lotion, paste, spray or drops suitable for administration to the skin, eye, ear, nose or genitalia.

For oral administration the pharmaceutical composition will be in the form of a tablet, capsule, powder, pellet, atroche, lozenge, syrup, liquid, or emulsion.

The pharmaceutical carrier employed may be, for example, either a solid or liquid. When the pharmaceutical composition is employed in the form of a solution or suspension, examples of appropriate pharmaceutical carriers or diluents include: for aqueous systems, water; for non-aqueous systems: ethanol, glycerin, propylene glycol, olive oil, corn oil, cottonseed oil, peanut oil, sesame oil, liquid paraffins, and mixtures thereof 'with water, for solid systems: lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid, kaolin and mannitol; and for aerosol systems: dichlorodifluoromethane, chlorotrifluoroethane and compressed carbon dioxide. Also, in addition to the pharmaceutical carrier or diluent, the instant compositions may include other ingredients such as stabilizers, antioxidants, preservatives, lubricants, suspending agents, viscosity modifiers and the -12- WO 94/25465 PCT/US94/04866 like, provided that the additional ingredients do not have a detrimental effect on the therapeutic action of the instant compositions. Similarly, the carrier or diluent may include time delay material well known to the art, such as glyceryl monostearate or glyceryl distearate alone or with a wax, ethylcellulose, hydroxypropylmethylcellulose, methylmethacrylate and the like.

A wide variety of pharmaceutical forms can be employed. Thus, if a solid carrier is used, the preparation can be tableted, placed in a hard gelatin capsule in powder or pellet form or in the form of a troche or lozenge. The amount of solid carrier will vary widely but preferably will be from about 25 mg to about 1 gram. If a liquid carrier is used, the preparation will be in the form of a syrup, emulsion, soft gelatin capsule, sterile injectable solution or suspension in an ampule or vial or nonaqueous liquid suspension. To obtain a stable water soluble dose form, a pharmaceutically acceptable salt of the compound of Formula I is dissolved in an aqueous solution of an organic or inorganic acid or base. If a soluble salt form is not available, the compound of Formula I may be dissolved in a suitable co-solvent or combinations thereof. Examples of such suitable cosolvents include, but are not limited to, alcohol, propylene glycol, polyethylene glycol 300, polysorbate glycerin and the like in concentrations ranging from 0-60% of the total volume.

It will be appreciated that the actual preferred dosages of the compounds used in the compositions of this invention will vary according to the particular complex being used, the particular composition formulated, the mode of administration and the particular site, host and disease being treated. It is expected that these compounds will be active in the concentration ranges of two commercial antiviral drugs, Cytovene (ganciclovir) and Zovirax (acyclovir). The latter is manufactured in 200 mg capsules with instructions for treating herpes simplex viral infections by taking one capsule every 4 hours, but not to exceed 5 capsules per day.

EXAMPLES

In the following synthetic examples, temperature is in degrees Centigrade Unless otherwise indicated, all of the starting materials were obtained from commercial sources. Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. These Examples are given to illustrate the invention, not to limit its scope.

Reference is made to the claims for what is reserved to the inventors hereunder.

-13- I- WO 94/25465 PCT/VS94/04866 ElXAMPLEI Preparation of 7-Acm1y-8-methyldioxolo [4.5-g I indolizino rl.2-b 1 quinolin-9(l 1W)- IA) 2-Methoxv-pvrrline 2-Pyrrolidone (850 g, 760 mL, 10 mol; Aldrich) was added dropwise, over a period of two hours, to a stirred solution of dimethyl sulfate (1260 g, 945 mL, 10 mol; Aldrich) under an argon atmosphere, causing the temperature to rise to 45*C. When addition was complete the clear mixture was stirred for I 6h at 60 0 C. It was then poured onto ice and saturated K2C03, and extracted with ether (2 X 1L). -1he combined organic phase was washed with brine, dried (Na2SO04), and solven i removed on the rotary evaporator, keeping the heating bath at 20*C. The residual liquid was distilled under vacuum into a chilled receiver to yield, after a small forerun 635 g (64%) of colorless liquid, b.p. 35 0 C 15 Torr: 1 NMR (400 MHz, CDCl3) 8 3,8 3H, OCH3), 3.65 2H, CH2-N), 2.45 2H, CH2-N), 2.1 (in, 2H, -CH2-).

1B) 7-Hvydroxy-8-ethoxvcarbonyl-2.3-dihvdro- 1H-indoli7zin-5-one The mixture of compound of Example 1 (100 g, 100 mL, 1 mol) and 1,3diethyl acetonedicarboxylate (202 g, 182 mL, 1.5 mol, Aldrich) alone with triethylamine (10 mQL was kept at room temperature for 1.5 weeks. The crystals formed were separated by filtration and washed with petroleum ether and ethyl ether to give off white solid 94 g m.p. 131OC: 1 NMR (400 MHz, CDCl3) 855.8 (s, 1H, olefin), 4.4 (mn, 3H, ethyl ester), 4.15 2H, CH2-NCO), 3.5 2H, CH2olefin), 2.25 (mn, 2H, 1.4 3H, ethyl ester).

1IC 7-HBydroxy-8-ethoxycarbonl-6-mnethvl-2.3-dihydO- I -noihi To a solution of compound of Example 1 (10 g, 45 inmol) in dry THF (500 mL) under an argon atmosphere was added NaH (2 g, 49 inmol, dispersion). The resulting mixture was stirred at room temperature for 10 mini.

Methyl iodide (2.8 mL, 45 minol) was added and mixture stirred at room temperature for 96h. Solvent was evaporated and residue purified by flash column chromatography (silica, 0-2% methanol: CH2Cl2) to give white solid 5.7 g (54%):1 NMR (400 MHz, CDCl3) 8 11.4 1H, OH), 4.4 (mn, 3H, ethyl ester), 4.15 2H1, CH2-NCO), 3.5 2H1, CH2-olefin), 2.25 (in, 2H, 2.01 3H, CH3), 1.4 311, ethyl ester) 1D) 7-Hvdroxy-8-carboxv-6-inethyl-2.3-dihydro- -14- WO 94/25465 WO 942S4~SI'MTUS94/04866 To a solution of compound of Example I1(C) (1.2 g, 5 mmol) in methanol inL), THF (20 mL) and H20 (20 inL) was added LiOH (1ig, 25 minol) and mixture stirred at room temperature for 56h. Solvent was removed in vacuum, The resulting mixture was diluted with H120 and acidified to (PH-5) with 3N RdI. The precipitated solid was filtered and washed with H20 and dried in vacuum to give tan solid 0.8 g 1 NMR (400 101-z, CD3OD) 8 4.23 (mn, 2H, CH2-NCO), 3.5 (t, 2H, CH2-olefmn), 2.35 (mn, 211, 1.95 3H1, CR3).

1E) 7-bxdrxy-6mtbI-2.3-dihdo-indolizin-5-one The compound of Example 1(D) (0.8 g, 3.75 inmol) and 2,4,6trichlorophenol (6 g) were heated at 220'C until evolution of carbon dioxide stopped.

The resulting mixture was cooled to room temperature and diluted with ethyl ether.

The precipitated solid was filtered and dried to give solid 0.62 g 1 NMR (400 MHz, CD3OD) 8 6.1 111, pyridyl), 4.23 (in, 2H1, CH2-NCO), 3.5 2H, CH2olefin), 2.35 (mn, 211, -CR2-, 1.95 3H, CH3).

1F) Trifluorometanesulfgnir, acid-6moty-i I .2.3.5-tetrahvdr-indoizin-7-I-e}i To the solution of compound of Example 1 (5 g, 30 inmol) in DMF (100 inL) was added triethylamine (12.6 inL, 90 nirol) alone with N-phenyltrifluoroinetanesulfonimide (16 g, 45 niiol). The resulting mixture was stirred at room temperature for I h. Solvent was removed in vacuum and residue purified by flash column chromatography (silica, 50-100% EtOAc: hexanes) to give tan solid 6.15 g 1 NMR (400 NMz, CDCl3) 8 6.15 111, pyridyl), 4.16 2H, -CH2), .13 211, 2.24 (in, 2H1, -CH2), 2.13 3H, CR3).

IG) 7( Btoyvnl-- To the solution of compound of Example 1 (6.15 g, 20 minol) in DMF (100 niL) was added triethylaniine (5.5 mL, 40 rinol) alone with n-butyl vinylether (10.3 mL, 80 minol). To the resulting mixture was added Pd(OAc)2 (0.27 g, 1.2 inrnol) alone with 1,3-bis(diphenyiphosphino)propane (0.49 g, 1.2 mmnol). TI"he resulting mixture was stirreed at 6000 for 5h. Solvent was removed in vacuum and residue purified, by flash column chromatography (silica, 30-60% EtOAc: hexane) to give oil 5 g 1 NWR (400 MHz, CDCl3) 8 6.15 111, pyridyl), 4.35 1H1, olefin), 4.16 (nm, 311, -CH2; olefine), 3.79 2H, CH12-O), 3.1 211, 2.2 (in, 5H, -CR3; 1.72 (in, 211, alkyl), 1.48 (in, 211, alkyl), 0.95 3H1, alkyl).

111) 7-Acetvl-methyl-2.3-dihvdro-IH-indolizin-5-7one, To the solution of compound of Example 1 (5 g, 20 iniol) in glacial acetic acid (10 niL) was added 3N HCI (3 m.L) anid the reaction mixture was stirred WO 94/25469 WO 94IS46~ CTiUS94/04866 at room temperature for lh. Solvent was removed in vacuum and residue resuspended in EtOAc, washed with 5% NaHCO3, NaCi, dried (Na2SO4). Solvent was removed in vacuum and residue purified by flash column chromatography (silica, 40-100% EtOAc: hexane and 0-5% methanol: CH2CI2) to give solid 2.3 g imp. 101-102'C.

H1) 6-Methvl-7-(2-methyl-[1I.31-dioxolan-2yl)-2.3-dihvdro- I HCl gas was bubled in to the solution of compound of Example 1 (2 g, 10.4 tumol) in ethelyneglycol (50 xnL) at 0 0 C. The resulting solution was alowed to warm-up to room temperature and stired at room temperature for 14h. The resulting mixture was poored in to the solution NH40H and ice. The mixture was extracted with CH2CI2, washed with H20, NaCi and dried (Na2SO4). Solvent was removed in vacuum and residue purified by flash column chromatography (silica, methanol: CH2C12) to give solid 2.13 g 1 NMR (400 MHz, CDCl3) 8 6.37 1H, pyridyl), 4.14 2H, 4.02 (in, 2H, ketal), 3.73 (in, 2H, ketal), 3.04 2H, 2.27 3H, -CH3), 2.16 (in, 2H, 1.61 3H, -CH3).

IJ) 1-Hydy-mcthvl-7-(2-nethI-rlI.31-dioxolan-2v1)-2.3-dihvdro IH- To the solution of diisopropylarnine (1.89 mL, 13.6 inmol) in THIF (20 mL) at -78'C was added n-butyllithium (5.4 inL, 13.6 minol) and the resulting mixture was stirred at -78*C for 10 mini. Tht solution of compound of Example 1(1) (2.13g, 9 minol) in THF (100 mL) was added via addition funnel and the resulting mixture was stirred at -78'C for 10 min Davis reagent (3.54 g, 18 minol) in THF (20 mL) was added at once and the resulting mixture was stirred at -78 0 C for 1h. Saturated solution of NH4Cl (20 mL) was added at -78'C and the resulting mixture was extracted Vith 0120I2. Aqueous layer was acidified with 3N HCl and extracted with with CH2Cl2. The combined organic fractions were washed with H20, brine and dried (Na2SO4). Solvent was removed in vacuum and residue purified by flash column chromatography (silica, 0-7% methanol: CH2CI12) to give foam 1. 16 g NMR (400 MHz, CDCi3) 8 6.66 1H, pyridyl), 5.21 (br s, 1H, -CH- OH), 4.14 (mn, 1H, 4.04 (br s, 1H, 3.99 (in, 3H, ketal, 3.73 (in, 2H, ketal), 2.46 (in, 1H, 2.27 3H, -CH3), 2.16 (in, 1Hi, 1.61 3H, -CH3).

1K) 6-Methvl-7-(2-methy]-r I .31-dioxolan-2vl)-2.3-dihvdro- I -indolizin-l To the solution of compound of Example 1 (1.1g, 4.3 inmol) in CH202 (100 inL) was added pyridiniuinchlorochromate (1.89 g, 8.7 inmol) and the resulting -16- WO 94/25465 WO 9415465 CT/US94/04866 mixture was stirred at room temperature for 12h. The mixture was diluted with C112C12 and the resulting residue was filtered through the bed of celite. Solvent was removed in vacuum and residue purified by flash column chromatography (silica, 0- 3% methanol: 011202) to give foam 0.8 g 1H1 NI4R (400 MHz, CDCl3): 8 7.20 1H1, pyridyl), 4.28 (br s, 2H1, 4.07 (br s, 2H, ketal), 3.73 (br s, 2H, ketal), 2.89 (br s, 2H, 2.42 3H, -CH3), 1.61 3H1, -0113).

1L) 7-Agtyl-8-mthldi!xolo 1 indolizino 1.2-b I quinolin-90 1 H)-one, To the solution of compound of Example I1(K) (0.l1g, 0.4 mmol) in toluene mL) was added arninopiperinal (73 mg, 0.4Ammol) alone with p-toluenesulfonicacid (2mg, catalyst). The resulting mixture was refluxed uner Dean-Stark trap for 12h. The mixture was cooled and diluted with hexane and Et2O. The precipitated tan solid was filtered and dried in vacuo to give ketal 65mg 1H NMR (400 MHz, CDCl3): 8 8.1 1H, 12quinolyl), 7.57 1H, 13-quinolyl), 7.48 111, 4-quinolyl), 7.14 111, 6-pyridyl), 6.18 2H, 0-0112-0), 5.19 2H, 11 4.09 (in, 2H, ketal), 3.84 (in, 2H1, ketal), 2.45 3H1, -0113), 1.72 3H, -CH3).

To the ketal above (65 mg, 0. 17 mmol) in glacial acetic acid (5 mL) was added 3N HCI (1 mL). The resulting mixture was heated at 70'C for lh. The mixture was diluted with H20 and extracted with 0112012. The oragnic layer was washed with brine and dried (Na2S04). Solvent was removed in vacuo to give yellow solid 28 mg 111 NMR (400 MHz, CDCI3+ CD3OD): 588.23 1H1, 12-quinolyl), 7.49 1H1, 13-quinolyl), 7.35 1H, 4-quinolyl), 7.19 1H1, 6pyridyl), 6.20 211, 0-0112-0), 5.23 211, I11-CH2-), 2.64 3H1, -CH3), 2.32 (s, 3H1, -0113); Anal. (CjqHl4N204. 0.5 H20) calcd.: C, 66.47; H, 4.40; N, 8.16 found: C, 66.41; H14.20; N, 8.14. -m.p >30000.

-17- WO 94/25465 WO 941546S CT/US94/04860 flXAMPLE2 E=atjon of 7-Acervl-I 2-dimethy)aminomethy)-8-methvldioxolo [4.5-g I indolizinc L1.2-t I quiflifl-9(1 Iffi-one.

2A) 1-(2-nitro-pherivI~etanone To the cooled 65% H{N03 (200 mL) was added 3,4-Methylenedioxyacetophenone (41 g, 0.25 mol; Aldrich) and the resulting mixture was stirred at 40*C for Ih with the evolution of heat. When the evolution of the heat ceased the mixture was poured into the ice. The water was decanted and the resulting gum was triturated with methanol. The product solidified and filtered to give solid 40g 1H- NMR (400 MHz, CDCl3): 8 7.55 111, aromatic), 6.76 1H, aromatic), 6.19 2H, O-CH2-O), 2.56 3H, CH3)#; m-p 1 12'C.

2B) 2-BroQmo- I -(2-nitro-henvljethano To the solution of compound of Example 2(A) (10 g, 47.8 mmol) in dioxane mL) was added dropwise the solution of bromine (2.5 mL, 48.3 mmol) in dioxane (100 mQL via the addition funnel. The resulting mixture was stirred at room temperature for 4h. Solvent was removed in vacuum and mixture was diluted with Et2O, washed with 5% NaHCO3, H20, brine and dried (Na2SO4). Solvent was removed in vacuum and residue purified by flash column chromatography (silica, 0- Et2O: hexane) to give lacrymatory solid 8g 111 NMR (400 MHz, CDCl3): 8 7.62 1H, aromatic), 6.84 IIH, aromatic), 6.22 2H, O-CH2-O), 4.23 2H, CH2Br).

2C) N-be-nzylidine-N.N-dimethv]-hvdrazine To the solution of benzaldehyde (5 g, 47 mmol; Aldrich) in ethanol (250 mL) at 10 0 C was added dropwise N,N-dimethylhydrazine (5.37 rnL, 70.6 niiol; Aldrich) in ethanol (50 mL). The resulting mixture was allowed to warm up to room temperature and was stirred at room temperature for 30 min, and then refluxed for 20h. Solvent was removed in vacuum and resulting mixture was diluted with extracted with Et2O. Organic layer was washed with brine and dried (Na2SO4).

Solvent was removed in vacuum to give light yellow oil 5.2 g IH NMR (400 MHz, CDCl3): 8 7.6 (in, 2H, aromatic), 7.3 (mn, 2H, aromatic), 7.2 (in, 3H, aromatic; CH=N), 2.95 6H, diinethylainino).

-18- WO 94/25465 WO 94I2~t~5IUS94/o4866 2D) 2-Dimethylamino- I -(2-ni=r-phenyDV-ethanone To the solution of compound of Example 2(B) (0.5 1.73 mmol) in CH3CN mL) was added compound of Example 2(C) (0.26 g, 1.73 mmol) and the resulting mixture was allowed to stand at room temperature for 24h. The product precipitated and filtered to give solid 126 mg 1 H NMR (400 Mffz, CDC13+CD3OD): 8 7.65 1H, aromatic), 7.19 1H, aromatic), 6.27 2H, 0-CH2-O), 4.67 2H, CH2N+(CH3)2), 3.09 6W, dimethylamino+).

2E) I -(2-Anmino-phenvl)-2-dimethylamino-etbanone To the solution of compound of Example 2(D) (0.126 g, 0.38 mmol) in methanol (5 mL) was added 1N WCI (3 mL) alone with Ni2B (200 mg) and the resulting mixture was heated at 60'C for 1h. The mixture was diluted with H-20 and extracted with EtOAc. The oraganic layer was washed with brine and dried (Na2SO4). Solvent was removed in vacuum to give solid 76 mg 1 1H NMR (400 MHz, CDCl3): 8 7.28 1H, aromatic), 6.45 (br s, 211, -NH2), 6.13 1H, aromatic), 5.9 2H, O-CH2-0), 3.52 2H, CHi2N(CH3)2), 2.34 6H.

dimethylamino).

2F) 7-Acetv-12-dimethylaminoamethvl-8-methvldipooo r4.5-g 1 indolizino [1.2b1 guinolin-M( Wh)ome Compound of Example 1(K) (71 mg, 0.28 mmol) and compound of Example 2(E) (71 mg; 0.31 mmol) were reacted following the procedure of Example 1(L).

Solvent was evaporated and residue -purified by flash column chromatography (silica, 0-10% methanol: CH2Cl2) to give ketal as yellor solid 25 mg IH NMR (400 MIHz, CDCl3): 8 7.61 1H, quinolyl), 7.49 III, quinolyl), 7.45 1WI pyridyl), 6.15 2H., 0-CH12-0), 5.23 2H, 11l-CW2), 4.07 (in, 211, ketal), 3.82 (mn, 2H, ketal), 3.80 2H, CW2N(CW3)2), 2.45 3H, -CH3), 2.29 6W, dimethylamino), 1.71 311, -CH3).

The ketal above was hydrolyzed following the procedure of Example 1(L).

The resulting residue was lyophilized. to give solid 12 mg 1 H NMR (400 MIHz, D20): 8 7.4 1H, quinolyl), 7.2 1H, quinolyl), 7.01 1W pyridt 6.3 2H, 0-CFW2-0), 5.18 2H, 1 1-CH2), 3.65 2H, CW2N(CH3)2), 2.9c% 6H, diniethylamino+)2.66 3H, 2.17 3H, -CH13).

-19- WO 94125465 I'MUS94/04866 EXAMPLE 3 Prepardano of 7-Acety]-12-hydroxvmethyl-8-methyldioxolo2 r4.5-g I indolizino [.1.2-b 1 auinolinl-9(1 1W-one.

3A) Acetic acid-2-(2-nitro-phenvP)-2-oxo--ethyI ester To the solution of compound of Example 2(B) (0.5 g, 1.73 mnmol) in DMF mL) was added sodium acetate (0.43 g, 5.19 mmol) and the resulting mixture was heated at 67*C for lh. The resulting mixture was diluted with H20 and extracted with EtOAc, washed with brine and dried (Na2SO4). The solution was triturated with. hexane and precipitated solid was filterd to give 0.32 g 1

H

NMR (400 MIz, CDCl3): 8 7.58 1H, aromatic), 7.84 111, aromatic), 6.2 (s, 2H, O-CH2-O), 4.92 2H, CH2-OAc), 2.06 3H, -CH3).

3B) Acetic acid-2 (2-a 'no-phenvI)-2-oxo-ethvl. ester Compound of Example 3(A) (0.3 g, 1.2 mmol) was reduced following the procedure of Example 2(E) to give foam 0.24 g 1 H NMR (400 MHz, CDCl3): 856.89 1H, aromatic), 6.43 (br s, 2H1, -NH2), 6.17 11H, aromatic), 5.91 2H, O-CH2-O), 5.17 2H, CH2-OAc), 2.23 3H, LCH3).

3C) 7-Acel-1I2-hydroxvrethvl-8-methyldioxo2lo [4.5-g I indoliino g.guinolini-9(I 1W-one Compound of Example 1(K) (50 mg, 0.2 mmol) and compound of Example 3(B) (50 mg; 0.22 mmol) were reacted following the procedure of Example 1(L).

The precipitated solid was filtered to give ketal 45 mg 1 H NMR (400 MHz, GDC13): 857.50 1H, quinolyl), 7.30 1H, quinolyl), 7.15 1W pyridyl), 6.20 (s, 2H, O-CW2-O), 5.5 2H, -CW2Ac), 5.30 2W, 1 1-CH2), 4.07 (in, 2H, ketal), 3.82-(m, 2H, ketal), 2.50 3H, -CW3), 2.20 3H, -CH3), 1.71 3H, -CH3).

The ketal above (40 mng, 0. 1 mmol) was hydrolyzed following the procedure of Example 1(L) to give solid 30mg 1 W NMR (400 MHz, DMSOd6): 57.50 1H, quinolyl), 7.40 11W, quinolyl), 7.20 1WI pyridyl), 6.25 2H, O-CH2-O), 5.4 2H, -CH2Ac), 5.17 2H, 11 -CH2), 2.55 311, -CH3), 2.20 3H, -CW3).

EXAMPLE4 W±-7-(1-Hydroxvethvl-8mehvdioxgoo 4 .5-gl1indolizino [1.2-bl1 uinoliin-M( I- 2M~ 4A) (±)-7-(l-J-ivdrxvethvl)-8-methvldioxolo r4.5-g 1 indolizino [1.2-b 1 quino-lin- 2(liaLIn To the solution of 7-Acetyl-8-methyldioxolo [4,5-g]I indolizino [1,2-b] WO 9412546S WO 942S465PCI1US94/04866 quinolin.-9(1 111)-one (2 mg, 5pinol) in a mixture of MeOI- (0.2 niL, CH2CI2 (0.6 mL) and THF (0.2 niL) was added a single portion of sodium borohydride (2 mg, pinol). After stirring at room temperature for 1.5h, the solvent was removed in vacuum. The resulting residue was treated with 10% aqueous N114CI (150 pL) and allowed to stand at 4 0 C overnight. The solid which was formed was collected by filtration, washed sparingly with 1120 and dried to give yellow solid 1.7 mng, IH NMR (400 MIHz, CDCI3): 6 8.2 111, 12-quinolyl), 7.4 III, 13-quinolyl), 7.12 1H-, 4-quinolyl), 7.1 1H, 6-pyridyl), 6.15 2H, 0-CH2-0), 5.2 211, 11l-CH2-), 4.89 (mn, 1H, CHOH), 2.3 3H-, Methyl) 1.5 3H1, aliphatic).

EXAMP1LE (±--[1(Amnoace'lox1etfl1-8-n-ethvjdioxolo r4.5-g 1 indolizino r] .2-b 1 Quinol u-90 1 11)-one Hvydrotri'fluroacetate.

5A) .±LL[[..I-ievehx~abnlaioaeylxltvl8 m=bYldi~xoQI.I.g 1 indolizino. [1.2-b 1 guinolin-9(1 1-)-one.

To a suspension of (t-butoxycarbonyl)glycine (2 mg, 10 pniol) in CH202 (2 inL) under an argon atmosphere was added 1,3-dicyclohexylcarbodmmnide (2 mg, pinol). After stirring at room temperature for 0.5 h, (±)-7-(1-Hydroxyethyl)-8methyldioxolo [A4,5-g]I indolizino [1,2-b quinolin-9(1 1H-)-one.(2 mng, 5 pinol) was added, followed by 1 mg of 4-diinethylaniinopyridine. The resulting mixture was stired at room temperature overnight, then was filtered. The filtrate was washed successively with 2.5% aqueous NaHC03 (2 mL), 0. 1 N HCl (2 niL) and H120 (2 mQL, dried (Na2504). The solid residue was purified by flash column chromatography (silica, 0-3% methanol: C112Cl2) to give the title compound 2.5 mg, 1H NMR (400 MHz, CDCl3): 6 8.2 111, 12-quinolyl), 7.4 11, 13quinolyl), 7.12 1H1, 4-quinolyl), 7.1 1H1, 6-pyridyl), 6.15 211, 0-CH12-0), 5.89 (mn, 111, CHOH), 5.2 211, 1 I-CH2-), 5.03 (br s, 1H1, NH),4.14-3.94 (in, 2H, C112-NCO), 2.3 311, Methyl), 1.44 911, t-Bu), 1.5 311, aliphatic).

L-L)_-L-F(AminoaZetvfloxyjethvll -8-inethvldioxolo 1 indolizino [1.2-b-1 QlflflQii- 9 (l 1H)-one Hydrotrifluroaette To a stirring suspension of 1, -Dimetylethoxy)carbonyl~arnino] acetyl]oxy]ethyl]-8-methyldioxolo [4,5-i indolizino, [1,2-b quinolin-9(1 111)-one (2.5mg, 4 pmol) in 1,3-dimethoxybenzene (2 inL) under an argon atmosphere was added trifluoroacetic acid (2 mL). After stirring for 1.5 h at room temperature, the mixture was concentrated under reduced pressure. The residue was dissolved in 1120, extracted with Et20, filtered and lyophilized to afford the title compound as a -21- WO 94/25465 PCT/US94/04866 pale yellow solid 1.8 mg, -1H NMR (400MHz, CD30D) 8 8.12 1H, 12quinolyl), 7.45 1H, 13-quinolyl), 7.13 1H, 4-quinolyl), 7.16 1H, 6-pyridyl), 6.15 2H, O-CH2-O), 5.89 1H, CHOH), 5.1 2H, 11-CH2-), 4.02 (br s, 2H, CH2N), 2.25 3H, Methyl), 1.4 3H, aliphatic).

EXAMPLE 6 Parcnteral Composition To prepare a parenteral pharmaceutical composition of this invention suitable for administration by injection, 100 mg of a water soluble salt of a compound of Formula I is mixed with 10 ml of 0.9% sterile saline, and the mixture is incorporated into a dosage unit form suitable for administration by injection.

EXAMPLE 7 Oin omposition To prepare an oral pharmaceutical composition of this invention, 100 mg of a compound of Formula I is mixed with 750 mg of lactose, and the mixture is incorporated into an oral dosage unit form, such as a hard gelatin capsule, which is suitable for oral administration.

Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.

e 9* e oa -22- -i i

Claims (10)

1. A method for treating viral infections comprising administering to an infected host in need thereof an effective amount of a compound of Formula 1, or a pharmaceutically acceptable salt thereof, alone or in combination with a carrier, diluent or excipient 0 NN CH3 R j wherein: R is 0, -OH, and OR 1 RI is OCOR 4 or OP(O)(OH)R 5 wherein: R 3 is -H or lower allkyl; R4 is -CR 3 R 6 R 7 -(CH 2 )nCH2R 7 (where -(CH2)nCH2COOH (where NT'4H(CH2)n 7 (where n and -NH(CH2)nCH2COOH (where ni 0-3); R 5 is OH or CH 2 NH 2 R 6 is H or the side chain of any naturally occuring ax-amino acid; -23- WOQ 94/2565 9CT/US94104866 +N-N R 7 is NR 9 R 1 0 I NO -No where X is any pharmaceutically acceptable anion; R 8 is lower alkyl; R 9 and RIO are independently selected from the group consisting of -H, -CI-6 alkyl, and R 9 and RI 0 taken together to form a membered saturated heterocyclic ring containing the nitrogen on which R 9 and R 1 0 are substituted; and RI 1 is -CH 2 R 12 wherein: -N 0 -N NCH, -OH R12 is -N(CH3)2

2. The method of claim I wherein said compound is selected from the group consisting of:

7-Acetyl-8-methyldioxolo[4,5-g]indolizino ,2-b quinoLin-9(1 11)-one; (±)-7-(l-Hydroxyethyl)-8-methyldioxolo[4,5-g]indolizino[1,2-b]quinolin- 9(1 11)-one; (±)-1-[1(Aminoacetyl)oxy]ethyl]-8-methyldioxolo[4,5-g]indolizino[1 ,2- b]quinolin-9(1 1H)-one hydrotrifluracetate; 7-Acetyl-12-dimethylaminomehy!;.g-xn.thyldioxolo indolizino [1,2-b]quinolin-9(1 1H)-one; and 7-Acetyl-12-hydroxylneh'iyl-8-methyldioxolo indolizino [1,2-b] quinolin-9(l 1H)-one. 3. Thv method of claim 2 wherein said compound is 7-acetyl-8- 1,2-b]quinolin-9(1 1H)-onz. 4. The method of claim 2 wherein said compound is 7-acetyl-12- dimethylamninomethyl-8-methyldioxolo indolizino 1,2-b]quinolin-9(l 1)- one. A compound of formula 1, or a pharmaceutically acceptable salt thereof, -24- WO 94/25465 WO 9425465PCT/US94104866 hI wherein: R is and OR 1 R 1 is OCOR 4 or OP(O)(OH)R 5 wherein;, R 3 is -11 or lower alkyl; R 4 is -CR 3 R 6 R 7 -(CH 2 )rCH 2 R 7 (where iS -(CH 2 )nCH 2 COOH (where -NR 9 R 10 -NH(CH 2 )nCH 2 R 7 (where n and -NH(CH 2 )nCH 2 COOH (where n 0-3); R 5 is OH or CH 2 )NH 2 R 6 is H or the side chain of any naturally occuring a-amino acid; where X is any pharmaceutically acceptable anion; WO 94/25465 PCT/US94/04866 +No X NI)- R 7 is NR 9 R' #where X is any pharmaceutically acceptable anion; R 8 is lower alkyl; R 9 and R 10 are independently selected from the group consisting of -H, -C1-6 alkyl, and R 9 and R 10 taken together to form a 5-7 membered saturated heterocyclic ring containing the nitrogen on which R 9 and R 1 0 are substituted; and R 1 1 is -CH 2 R 12 wherein: -N 0, -N NCH -OH R12 is-N(CH3)2 6. The compound of claim 5 wherein said compound is selected from the group consisting of: 7-Acetyl-8-methyldioxolo[4,5-g indolizino[ 1,2-bquinolin-9(1 1H)-one; (±)-7-(1-Hydroxyethyl)-8-methyldioxolo[4,5-g]indolizino[ 1,2-b]quinolin- 9(1 1H)-one; (±)-7-[(Aminoacetyl)oxy ethyl]-8-methyldioxolo[4,5-g]indolizino[1,2- b]quinolin-9(1 1H)-one hydrotrifluracetate; 7-Acetyl-12-dimethylaniinomethyl-8-methyldioxolo indolizino [1,2-b]quinoin-9(1 1H)-one; and 7-Acetyl- 12-hydroxymethyl-8-methyldioxolo indolizino [1,2-b] quinolin-9(l 1)-one. 7. The compound of claim 6 wherein said compound is 7-acetyl-8- methyldioxolo[4,5g]indolizino[1 ,2-b]quinolin-9(1 11)-one.

8. The compound of claim 6 wherein said compound is 7-acetyl-12- dimethylaminomethyl-8-methyldioxolo indolizino [1,2-bquinoiin-9( 1IH)- one.

9. A formulation comprising a compound of claim 5 in admixture with a carrier or excipient. The formulation of claim 9 wherein said carrier or excipient is a pharmaceutically acceptable carrier or excipien. -26- WO 94/25465 PCT/US94/04866

11. The method of claim 1 wherein the viral infection is caused by a herpesvirus.

12. The method of claim 11 wherein said virus is herpes simplex type 1 and said infected host is a mammal.

13. The method of claim 11 wherein said virus is herpes simplex type 2 and said infected host is a mammal.

14. The method of claim 1 wherein said viral infection is caused by cytomegalovirus and said infected host is a mammal. The method of claim 1 wherein said viral infection is caused by varicella zoster virus and said infected host is a mammal.

16. The compounds of claim 5 and ccmpositions of claim 9 substantially as hereinbefore described with reference to the Examples. DATED this NINETEENTH day of SEPTEMBER 1997 SmithKline Beecham Corporation By DAVIES COLLISON CAVE Patent Attorneys for the applicant 0 *e *e *e t -27- W INTERNATIONAL SEARCH REPORT Intemnational application No. PCTIUS94/04M6 A. CLASSIFICATION OF SUBJECT MAT[TER :C07D 487/22; A61K 31/475 US CL :546/48; 514/80, 283; A61K 31/475 According to International Patent Classification (IPC) or to both national classification and IPC B. FIELDS SEARCHED Minimum documentation searched (classificatioai system followed by classification symbols) U.S. ,546/48; 514.80, 283; A61K 31/475 Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched Electronic data base consulted during the international search (name of data base and, where. practicable, search terms used) CAS online structure search C. DOCUMENTS CONSIDERED TO BE RELEVANT Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No. Y WO, A, 92/07856 (ALLAUDEEN et at.) 14 May 1992. See 1-15 claims 1, 2, 7-9, 40-42, 47-52, 73-6 where Y is CH ,X is C-6 and R is lower alkyl. Y US, A, 5,225,404 (GIOVANNELLA et al.) 06 July 1992. See 1-8 claim 1. Y US, A, 4,981,968 (WALL et at.) 01 January 1991. See 1-15 column 3, lines 48-50. A US, A, 4,914,205 (SAWADA et al.) 03 April 1990. 1-15 Further documents are listed in the continuation of Box C. 1 See patent family annex. Special categories of ciltd documnts: r later document published after the iternational ists date or priotity A downetdetnig te gnerl stte r te at wich~ ~date and not in confliciwith the application but cited nidersuWAn the t'A' donurin th tnrlsaeo h r hc nntcniee principle or theory underlying the invention E ealie doumen puliaed o oraftr te inerntioal flin dae docunxat or partcular relevance; the claimed Invewwt, cannot be arler ocumnt ublshedon r aterthe ntenatonalrilng ateconsiered novel or cannot be considered to involve so, iinveotiveulep -L document which may thtrow doubts on priority clauim(s) or which 6 when the document is taken al,4oc cited to establish the publication date or another citation or other doumn 'fpriua eeac;tecie neto antb specialonidre reoo (anvecoid)"i an inventive step when the document is document referring to an oral disclosure. use. exthibition or other combined with one or more other such documents, such combination means being obvious to a person skilled in the ant .P document published prir~rto the internuttional ling date but Later Itn document member of the sme patent family the priority date &*,aod Date of the actual completion of the interniptional search Date of mailing of the internatio-na search At 08 JUNE 1994 AUG 05 1994 Name and mailing address of the ISA/US Authorized officer Commissioner of Patents and Tradenmarkcs Box PCT DC AS j Washington, D.C. 20231 Facsimile No. (703) 305-3230 Telephone No.- (703) 308-123 Form PCT/ISA/210 (second sheet)(July 1992)* INTERNATIONAL SEARCH REPORT liiternational application No. PCT/US94/04866 C (Continuation). DOCUMENTS CONSIDERED TO BE RELEVANT Category* Citation of document, with indication, where appropriate, of thc relevant passages 4 4 Relevant to claim No. 1-15 Chemical Abstracts, Vol. 84, issued 1976, Atherton et al., "Interferon Induction by Viruses and Polynucleotides" See page 428, abstract No. 103727f. J. Gen. Virol. 1975, Pt 3. 297-3054. Chemical Abstracts, Vol. 77 issued 1972, Hlorwitz et al. "Camptothecin, Mechanism of Inhibition of Adenovirus Formation". See page 7079, col. 1, Abstract No. 70781u, Virology Vol. 48, No. 3 pages 690-8 (1972). JP, A, 51-91297 (Nippon Chemifar) 08 October 1976. See abstract. 11-15 Form PCT/15A/210 (continuation of second shtct)(uIY 1992)* I INTERNATIONAL SEARCH REPORT International application No, PCT/US94/04866 Box I Observations where certain claims were found unsearchable (Continuation of item 1 of first sheet) This international report has not been established in respect of certain claims under Article 17(2)(a) for the following reasons: 1. j' Claims Nos.: because they relate to subject matter not required to be searched by this Authority, namely: 2. F[ Claims Nos.: because they relate to parts of the international application that do not comply with the prescribed requirements to such an extent that no meaningful international search can be carried out, specifically: 3. D- Claims Nos.: because they are dependent claims and are not drafted in accordance with the second and third sentences of Rule 6.4(a). Box II Observations where unity of invention is lacking (Continuation of item 2 of first sheet) This International Searching Authority found multiple inventions in this international application, as follows: Please See Extra Sheet. 1. As all required additional search fees were timely paid by the applicant, this international search report covers all searchable claims. 2. E As al searchable claims could be searched without effort justifying an additional fee, this Authority did not invite payment of any additional fee. As only some of the required additional search fees were timely paid by the applicant, this international search report covers only those claims for which fees were paid, specifically claims Nos.: 4. No required additional search fees were timely paid by the applicant. Consequently, this international search report is restricted to the invention first mentioned in the claims; it is covered by claims Nos.: 1-15(part of 1,5,9-15) Remark on Protest The additional search fees were accompanied by the applicant's protest. No protest accompanied the payment of additional search fees. Form PCT/ISA/ (ontinuation of Form PCT/ISA/210 (continuation of first sheet(l))(July 1992)* I e I INTERNATIONAL SEARCHI kEEPORT International application No. PCT/US94/04866 BOX 11, OBSERVATIONS WHERE UNITY OF INVENTION WAS LACKING This ISA found multiple inventions as follows: I. Claims 1, 5, 9-15 (part of each) and 2-4, 6-8, drawn to non- morpholino, non-piparazino compounds and antiviral use. II. Claims 1, 5, 9-15 (part of each) drawn to Morpholinr compounds, use. Ill. Claims 1, 5, 9-I5 (part of each) drawn to Piperazinc compounds, use. PCT Rule 13.3 permits a lack of unity holding where joined in a single claim. Applicants did not disclose Groups 11 and Ill in their first priority application, as having the "special technical feature" in common with I. The technical feature is shared with the art, i.e. not "special". I Form PCTIISAI21O (extra sheet)(July 1992)*