ANTHELMINΗC β-KETOAMIDES BACKGROUND OF THE INVENTION The diseases or groups of diseases described generally as helminthiasis are due to infection of the animal with parasitic worms known as helminths. Helminthiasis and helminthosis are prevalent and may lead to serious economic problems in valuable domestic warm-blooded animals such as sheep, swine, cattle, goats, dogs, cats, horses, poultry and man. Among the helminths, the groups of worms known as nematodes. trematodes and cestodes cause widespread and often-times serious infections in various species of animals including man. The most common genera of nematodes, trematodes and cestodes infecting the animals referred to above are Dictyocaulus, Haemonchus, Trichostrongylus, Ostertagia, Nematodirus, Cooperia, Bunostomum, Oesophagostomu , Chabertia, Strongyloides, Trichuris, Fasciola, Dicrocoelium, Enterobius, Ascaris, Toxascaris, Toxocara, Ascaridia, Capillaria, Heterakis, Ancylostoma- Uncinaria. Dirofilaria- Onchocerca, Taenia, Moniezia. Dipylidium, Metastrongylus, Triodontophorus, Macracanthorhynchus, Hyostrongylus, and Strongylus. Some of these genera attack primarily the intestinal tract while others inhabit the stomach, lungs, liver and subcutaneous tissues. The parasitic infections causing helminthiasis and helminthosis lead to anemia, malnutrition, weakness, weight loss, unthriftiness, severe damage to the gastrointestinal tract wall and, if left to run their course, may result in death of the infected animals.

INFORMATION DISCLOSURE STATEMENT Anthelmintic compounds which also contain a ketoamide structure are disclosed in US

Patent 3,746,765, and in Swedish patent 592,048.

SUMMARY OF THE INVENTION This invention pertains to a new method for killing and controlling worms (Helminths), and new formulations for killing and controlling worms in animals, and new chemical compounds. The invention is more particularly directed to a new method for killing and controlling parasitic worms in animals with novel β-ketoamides, and to new anthelmintic formulations comprising the same.

The anthelmintic β-ketoamides have the general structure of Formula I. This invention encompasses: 1) A compound of formula I

(Formula I)

wherein each variable may be independent of the other variable unless indicated otherwise; the C-C bonds, a. b, and c, are single or double bonds; either a or b are double bonds; X is either -NH- or -0-;

R- is =0. -OH, -OCH ₃ or -NH ₂ ,

R- is =0 when b is a double bond and when a is a single bond, when R _t is =0 then R ₃ is -OH, -OCH ₃ or -NH ₂ ; R ₃ is =0, -OH, -OCH ₃ or -NH ₂ , R ₃ is =0 when a is a double bond and when b is a single bond, when R ₃ is =0 then R, is -OH, -OCH ₃ or -NH ₂ ;

R ₂ is

*2-l ^■ 2-2

2-3

R,

-GHoCB-j

R _2.] . R _2.2 and R ₂ . ₃ are, each independently: H, Halogen, CF ₃ , CN, N0 ₂ , OCH ₃ , NH ₂ , CI- C5 alkyl. or C,-C ₅ alkoxy; n is 1 to 3, R ₄ and R ₇ taken together are, -0-, -CH ₂ -,

R ₅ is H, Halogen, or C,-C ₅ alkyl;

Re is H, Halogen, or C--C ₅ alkyl; or

R ₅ and Rg taken together are

wherein

R ₅ is Halogen.

2) A pharmaceutical composition consisting essentially of a pharmaceutically acceptable carrier and an effective amount of a compound of formula I and 3) A method of killing or controlling parasitic worms in humans and warm blooded domestic animals which comprises administering a therapeutic or prophylactic dosage of a compound of formula I. The use of a compound of formula I or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the killing or controlling parasitic worms in humans and warm blooded domestic animals- Pharmacologically acceptable salts of Formula I are also included as one aspect of this invention. The anthelmintic agents of this invention will find their primary use in the treatment and/or prevention of helminth parasitism in valuable warm-blooded domesticated animals such

as sheep, cattle, horses, dogs, swine, goats and poultry; they are also effective in treatment that occurs in other warm blooded animals including man.

DETAILED DESCRIPTION OF THE INVENTION The compounds of this invention all possess a β-ketoamide (BKA) template. This β- ketoamide (BKA) template is then joined to one of four different components. The four compo¬ nents are: oxynorbomene BKAs (CHART A), norbomene BKAs (CHART B), tetracyclic BKAs (CHART C) and fulvene BKAs (CHART D). The compounds of this invention can be made in accordance with the processes illustrated in and the reactions related to CHARTS A, B, C, and D. The reactions are described in general teims immediately below, the detailed preparations are found at the end of this section, followed immediately by the CHARTS, which show the compound structures.

CHART A Reactions of Chart A (oxynorbomene β-ketoamides) Step 1. The starting ketone, 1 ,3-Cyclopentenedione, and furan (used as both reactant and solvent) are refluxed for 3 to 5 days to produce the Diels- Alder adduct, Figure Al. The compound of Figure Al is stirred with various arylisocyanates and benzoylisocyanates in tetrahydrofuran at room temperature in the presence of an organic base to prepare the oxynorbomene BKAs of CHART A. Steps 2 through 4, below, provide several examples.

Step 2. The compound of Figure Al and 4-bromophenylisocyanate gives Figure A2. From step 2. reactions may proceed in any of several directions; as shown by the following series of reactions.

Step 3(a). series (a). Bromine is added to the compound of Figure A2 in CCL, to give the dibromo BKA, Figure A3(a).

Step 4(a), series (a). The reactions of Step 3(a) results in a mixture of cis-endo-, cis- exo-, and -rα/w-dibromo isomers which can be subsequently dehydrobrominated (KOH/PTC- /EtOH/25°C/10 days) in Step 4(a) to give the compounds of Figure A4(a).

Step 3(b), series (b). The compound of Figure A2 is treated with dibromoformaldoxime and silver nitrate, to give the bromoisoxazoline, A3(b).

Step 3(c), series (c). Hydrogenation of the compound of Figure A2 produces Figure A3(c).

Step 3(d), series (d). Unlike an ordinary alcohol, reaction of the compounds of Figure A2 with diazomethane readily produces the vinylogous ester, Figure A3(d).

Step 4(d), series (d). The compounds of Figure A3(d), the vinylogous ester, reacts with ammonia to form the vinylogous amide, Figure A4(d). CHART B

Reactions of Chart B (norbomene β-ketoamides)

Step 1. The Diels- Alder reaction between freshly distilled cyclopentadiene and maleic anhydride (Et _j O/ZS") produces the endo-adάuc the compound represented by Figure B I endo- not shown in figure).

Step 2. Reaction of the compound of Figure B 1 with ethyl lithioacetate gives Figure B2 as a mixture of stereoisomers.

Step 3- The compound represented by Figure B2 is converted to the ester, Figure B3. by treating it with carbonyl dϋmidazole and potassium tert-butoxide in tetrahydrofuran at reflux. Pure endo isomer of the compound of Figure B3 was produced by trituration with and partial recrystallization from boiling hexane. Step 4. The norbomene BKAs of CHART B are then prepared by simply combining the compound of Figure B3 with an aromatic amine in toluene and heating the mixture under re¬ flux. For example, the compound of Figure B4, is prepared from the ester, Figure B3, and 2- fluoroaniline.

Additional reactions can be performed on the norbomene BKAs themselves. They can be brominated and hydrogenated. The enolic hydroxyl group found in these BKAs, similar to the oxynorbomene BKAs, is very acidic and is best viewed as a vinylogous acid. They can be treated with diazomethane, to give the vinylogous ester. Treatment of the vinylogous ester with methanolic ammonia forms the vinylogous amide.

CHART C Reactions of Chart C (tetracyclic β-ketoamides)

Step 1. Cyclooctatetraene and maleic anhydride are heated under reflux, in toluene, to give the anhydride. Figure Cl.

Step 2. Reaction of the anhydride. Figure Cl, with ethyl lithioacetate in tetrahydrofuran at -78°C gives the β-ketoester-acid. Figure C2. Step 3. The β-ketoester-acid, Figure C2, is converted to the ester. Figure C3, by heating it with carbonyl diimidazole and potassium tert-butoxide in tetrahydrofuran at reflux-

Step 4. The ester. Figure C3, is heated with an aromatic amine such as 4-bromoaniline in toluene at reflux to obtain the compounds of Figure C4.

Other tetracyclic β-ketoamides are similarly prepared. CHART D

Reactions of Chart D (fulvene β-ketoamides) Step L Figure D-l, 6,6-dimethylfulvene, is prepared by stirring a methanol solution of freshly distilled cyclopentadiene with acetone in the presence of pyrrolidine.

Step 2(a), Series (a)- The compound of Figure D2(a) is obtained as a 2:1 mixture of exo- and emio-isomers by refluxing the compound of Figure DI and maleic anhydride in ether overnight.

Step 3(a), Series (a). The compound of Figure D2(a) is treated with ethyl lithioacetate at -78°C to give the β-ketoester-acid. Figure D3(a), as a yellow oil containing several impurities.

Step 4(a). Series (a). Without purification, the β-ketoester. Figure D3(a) is converted to the cyclized ester. Figure D4(a), by heating it with carbonyldiimidazole and potassium tert- butoxide in tetrahydrofuran.

Step 5(a), Series (a). The cyclized ester, Figure D4(a), is reacted with aryl amines to produce the BKAs. One BKA is the compound of Figure D5(a), produced by heating the ester. Figure D4(a) with 4-bromoaniline in toluene under reflux to give the compound of Figure D5(a).

An alternate strategy may also be used. (Chart D, page 1) Step 2(b), Series (b). Stirring the compound of Figure DI, the fulvene, with cyclopen- tenedione in ether at 25°C for six days gives the compound of Figure D2(b).

Step 3(b), Series (b). Treating the compound of Figure D2(b) with an isocyanate such as 4-bromophenylisocyanate under the same reaction conditions used for the oxynorbomenes (CHART A), gives the compound of Figure D3(b). Following the general procedures from above, specific compounds can be prepared. At the end of this section detailed preparations/examples describe how to prepare various β- ketoamides of the invention. The detailed procedures are followed by CHARTS and TABLES.

The utility of the compounds of this invention are demonstrated by their activity in biological screens. The results of two screens, or assays, can be found in Table 1. The left column of TABLE 1 identifies the compounds according to the EXAMPLE number. The EXAMPLE number corresponds to the EXAMPLE numbers in the detailed synthesis below. The other columns of TABLE 1 show the results of the two assays which are described below.

ASSAY No. 1 This assay uses cultures of Caenorhabditis elegans maintained in a culture grown in McArtney bottles at 20°C in 10cm ³ of a thymidine-fortified broth containing 10 g Bactotryptone, 5 g Oxoid yeast extract, 10 mg cysteine and 5 mg thymidine dm ^"3 . Worms for the test are washed from 4-8 day old cultures. Test medium was made by adding 10 cm ³ of well-grown Escherichia coli (3-5 days) to 100 cm ³ M9 buffer plus 5 mg ampicillin and 10 000 Units nystatin. (The M9 buffer is described in Brenner, S. Genetics, 1974, Vol. 77, p. 71.) After 2 hours at room temperature, 2 cm ³ of the test medium was placed in each well of a Sterilin Repli Dish (Catalogue number 103) and 10 υl of dimethyl sulphoxide solution of chemical added to each well. One drop of worm suspension containing 10-20 worms of mixed age was added to each well and the plates were incubated at 20°C for 7 days.

After 7 days the plates were viewed under a binocular microscope and assessed for number of worms present A dose level of 50 parts per million (PPM) was chosen for the screen. A score of any number greater than zero suggests activity. A score of 5 means that no

live worms were present at the time of scoring. Assay No. 1 is completely described in: K-

Simpkin and G. Coles. J. Chem. Tech. Biotechnol. (1981), Vol. 31, p. 66-69.

ASSAY No. 2 This assay uses jirds, Meriones unguiclatus, infected with both

Haemonchus contortus and Trichostrongylus colubriformis. Female jirds weighing 30-35 g were obtained and put on a diet of commercial rodent chow containing 0.02 percent hydrocortisone for the duration of the study. All jirds were inoculated with approximately 1,000 exsheathed infective larvae each of H. contortus and T. colubriformis 5 days after the animals had been placed on medicated feed. Treatments were made 10 days post inoculation (PI). On day 13 PI,

3 days post treatment the jirds were killed, their stomachs and small intestines removed and examined for worms. The percent reduction in treated relative to the controls is provided in the

Table. Assay No. 2 is completely described in: G. A. Conder, et- al., J. Parasitology, Vol.

77(4) (1991), pi 621-623.

The preferred compounds of this invention are:

EXAMPLE (AX12) 8-Bromo-3-hydroxy-5-oxo-10-oxa- -tricyclo[5.2.1.0 ^2,6 ]dec-3,8-diene-4-carboxyIic acid, 4-bromophenylamide;

EXAMPLE (BX5) 3-Hydroxy-5-oxo-tricyclo[5.2.1.(F]deca-3,8-diene-^-carboxyli c acid,

4-bromophenylamide;

EXAMPLE (BX7) S-Hydroxy-S-oxo-tricyclofS^.l.C^^deca-S.S-diene-^carboxylic acid,

4-trifluoromethylphenylamide; EXAMPLE (BX12) 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ² - ⁶ ]deca-3,8-diene-4-carboxylic acid,

3,5-bis-trifluoromemylphenylamide;

EXAMPLE (BX22) S-Hydroxy-S-Oxo-i-ricyclo ^.l.O^deca-S.S-diene^-caT røxyUc acid,

7-bromo-9H-fluoren-2-ylamide;

EXAMPLE (BX24) S-Hydroxy-S-oxo-tricycloCS^-l.O^deca-S.S-diene-^-carboxylic acid, biphenyI-4-ylamide;

EXAMPLE (BX26) 2-(N-(4-trifluoromethylphenyI)carbamoyl)-5H,6H-dibromo* H,7H- methano-3-hydroxy-inden- 1-one;

EXAMPLE (BX27) 3-Hydroxy-5-oxo-tricyclo[5.2.1.(F]dec-3-ene-*4-carboxylic acid

4-bromophenyIamide; EXAMPLE (CXI) (2a-alpha., 3.beta., 3a.beta., 6a.beta., 7.beta., 7a-alpha-)-N-(4- bromophenyl)-2a,3a-4,6a,7,7a-hexahydro-6-hydroxy-4-oxo-3,7-E theno-3H-cyclobut[f]mdene-5- carboxamide,

EXAMPLE (CX6) (2a.alpha., 3.beta., 3a.beta., 6a.beta-, 7.beta., 7a.alpha.)-N-[[2-methyl-4-

(4-trifIuoromethyI)phenoxy]phenyl]-2a,3a,4,6a,7,7a-hexahy dro-6-hydroxy-4-oxo-3,7-etheno-3H- cyclobut[f]indene-5-carboxamide.

EXAMPLE (DX1) 3-Hydroxy-10-isopropylidene-5-oxo-tricyclot5.2.1.0 ² -°]deca-3,8-diene-

-9-

-4-carboxylic acid, 4-bromophenylamide;

EXAMPLE (DX2) 3-Hydroxy-10-isopropylidene-5-oxo-tricyclo[5.2. l.O^deca-S-β-diene-

-4-carboxylic acid, 4-trifluoromethylphenylamide.

Some of the compounds of this invention are useful as intermediates in the synthesis of active compounds. Some compounds may be both active and useful as an intermediate. Some of the useful intermediates are:

BX2 3-Ethoxycarbonylacetyl-bicyclo[2.2.1]hept-5-ene-2-carboxylic acid; BX4 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ² - ⁶ ]deca-3,8-diene*4-carboxylic acid, ethyl ester; BX29 S-Methoxy-S-oxo-tricyclotS^.l.C -^deca-S.δ-diene^-carboxylic acid, ethyl ester, CX7 8-(2-ethoxycarbonyl-acetyl)-tricyclo[4.2.2.0 2,5]deca-3,9-diene-7-carboxylic acid;

DX6 3-(2-ethoxycarbonyl-acetyl)-7-isopropylidene-bicyclo[2.2.1 ]hept-5-ene-2-carboxylic acid; DX7 3-hydroxy-10-isopropylidene-5-oxo-tricyclo[5.2.1.0 2,6]deca-3,8-diene-4-carboxylic acid ethyl ester. The compounds of this invention are identified in two ways: by the descriptive name and reference to labeled structures contained in appropriate charts and tables. Occasionally the proper stereochemistry is also represented in the charts. In this document the parenthetical term (C _n -C _m ) is inclusive such that a compound of (C-- ) would include compounds of one to 8 carbons and their isomeric forms. The various carbon moieties are defined as follows: Alkyl refers to an aliphatic hydrocarbon radical and includes branched or unbranched forms such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, isopentyl, n- hexyl, isohexyl, and n-octyl. Halogen or halo is fluoro, chloro, bromo, iodo or trifluoromethyl. THF is tetrahydrofuran. EtOH is ethanol. EtOAc is ethyl acetate. EtjO is diethyl ether. KOH is potassium hydroxide. PTC is Phase Transer Catalyst, in these reactions the PTC usually use is benzyltriethylammonium chloride. C is carbon. H is hydrogea 0 is oxygen. OH is hydroxy. =0 is carbonyl. NMR is Nuclear Magnetic Resonance. PPM is parts per million. ° is degree centigrade. L is liter. mL is milliliter. M or mol is mole. mM is milliMole. g is gram, mg is milligram, min is minute.

The compounds of this invention can be used as the pure compounds or as mixtures of pure compounds but for practical reasons the compounds are preferably formulated as anthelmintic compositions and administered as a single or multiple dose, alone or in combination with other anthelmintics (e.g. avermectins, benzimidazoles, levamisole,

praziquantel, etc.). For example, aqueous or oil suspensions can be administered orally, or the compounds can be formulated with a solid carrier for feeding- Furthermore, an oil suspension can be converted into an aqueous emulsion by mixing with water and injecting the emulsion intramuscularly, subcutaneously or into the peritoneal cavity. In addition, the active compound(s) can be administered topically to the animal in a conventional pour-on formulation. Pure compounds, mixtures of the active compounds, or combinations thereof with a solid carrier can be administered in the animal's food, or administered in the form of tablets, pills, boluses, wafers, pastes, and other conventional unit dosage forms, as well as sustained release dosage forms which deliver the active compound over an extended period of days, weeks or months. All of these various forms of the active compounds of this invention can be prepared using physiologically acceptable carriers and known methods of formulation and manufacture.

Representative solid carriers conveniently available and satisfactory for physiologically acceptable, unit dosage formulations include com starch, powdered lactose, powdered sucrose, talc, stearic acid, magnesium stearate, finely divided bentonite, and the like. The active agent can be mixed with a carrier in varying proportions from, for example, about 0.001 percent by weight in animal feed to about 90 or 95 percent or more in a pill or capsule. In the latter form, one might use no more carrier than sufficient to bind the particles of active compound.

In general, the compounds can be formulated in stable powders or granules for mixing in an amount of feed for a single feeding or enough feed for one day and thus obtain therapeutic efficacy without complication. It is the prepared and stored feeds or feed premixes that require care. A recommended practice is to coat a granular formulation to protect and preserve the active ingredient- A prepared sheep-feed containing about 0.2 percent of the active compound will provide a dosage of about 100 mg per kg body weight for each 100 lb sheep in its daily ration.

A solid diluent carrier need not be a homogeneous entity, but mixtures of different diluent carriers can include small proportions of adjuvants such as water; alcohols; protein solutions and suspensions like skimmed milk; edible oils; solutions, e.g., syrups; and organic adjuvants such as propylene glycols, sorbitol, glycerol, diethyl carbonate, and the like. The solid carrier formulations of the inventions are conveniently prepared in unit dosage forms, to facilitate administration to animals. Accordingly, several large boluses (about 20 g weight) amounting to about 54 g of active compound would be required for a single dosage to a 900 lb horse at a dosage rate of 50 mg/kg of body weight. Similarly, a 60 lb lamb at a dosage rate of 100 mg/kg of body weight would require a pill, capsule, or bolus containing about 2.7 g of active compound. A small dog, on the other hand, weighing about 20 lbs. would require a total dosage of about 225 mg at a dosage rate of 25 mg kg of body weight The solid, unit

dosage forms can be conveniently prepared in various sizes and concentrations of active ingredient to accommodate treatment of the various sizes of animals that are parasitized by worms.

Liquid formulations can also be used. Representative liquid formulations include aqueous (including isotonic saline) suspensions, oil solutions and suspensions, and oil in water emulsions. Aqueous suspensions are obtained by dispersing the active compound in water, preferably including a suitable surface-active dispersing agent such as canonic, anionic, or non¬ ionic surface-active agents. Representative suitable ones are polyoxyalkylene derivatives of fatty alcohols and of sorbitan esters, and glycerol and sorbitan esters of fatty acids. Various dispersing or suspending agents can be included and representative ones are synthetic and natural gums, tragacanth. acacia, alginate, dextran, gelatin, sodium carboxymethylcellulose, methylcellulose, sodium polyvinylpyrrolidone, and the like. The proportion of the active compound in the aqueous suspensions of the invention can vary from about 1 percent to about 20 percent or more. Oil solutions are prepared by mixing the active compound and an oil, e.g. an edible oil such as cottonseed oil, peanut oil, coconut oil, modified soybean oil, and sesame oil. Usually, solubility in oil will be limited and oil suspensions can be prepared by mixing additional finely divided compound in the oil.

Oil in water emulsions are prepared by mixing and dispersing an oil solution or suspension of the active compound in water preferably aided by surface-active agents and dispersing or suspending agents as indicated above.

In general, the formulations of this invention are administered to animals so as to achieve therapeutic or prophylactic levels of the active compound. Effective dosages of various compounds are contemplated, e.g., in the range of 1 to 75 mg/kg of body weight. In other animals, and for other kinds of parasitic worms, definitive dosages can be proposed. Contemplated are dosage rates of about 1 mg to about 800 mg/kg of body weight. A preferred, contemplated range of dosage rates is from about 1 mg to about 50 mg/kg of body weight. In this regard, it should be noted that the concentration of active compound in the formulation selected for administration is in many situations not critical. One can administer a larger quantity of a formulation having a relatively low concentration and achieve the same therapeutic or prophylactic dosage as a relatively small quantity of a relatively more concentrated formulation. More frequent small dosages will likewise give results comparable to one large dose. One can also administer a sustained release dosage system (protracted delivery formulation) so as to provide therapeutic and/or prophylactic dosage amounts over an extended period. Unit dosage forms in accordance with this invention can have anywhere from less than 1 mg to 500 g of active compound per unit

Although the anthelmintic agents of this invention will find their primary use in the treatment and/or prevention of helminth parasitism in valuable warm-blooded domesticated animals such as sheep, cattle, horses, dogs, swine, goats and poultry- they are also effective in treatment that occurs in other warm blooded animals including man. The optimum amount to be employed for best results will, of course, depend upon the particular compound employed, species of animal to be treated, the regimen of treatment and the type and severity of helminth infection. Generally good results are obtained with compounds of Formula I by the oral or parenteral route of administration of about 1 to 300 mg/kg of animal bodyweight (such total dose being given at one time, in a protracted manner or in divided doses over a short period of time such as 1-4 days). The technique for administering these materials to animals are known to those skilled in the veterinary and medical fields.

It is contemplated that the compounds of Formula I can be used to treat various helminth diseases in humans, including those caused by Ascaris, Enterobius, Ancylostoma, Trichuris, Strongyloides, Fasciola, Taenia, and/or Onchocerca or other filariae at a dose of from 1 mg/kg to 300 mg/kg of body weight upon oral and/or parenteral administration.

Below are detailed preparations and examples that describe how to prepare various β- ketoamids of the invention- The procedures are to be construed as merely illustrative, and not limitations of the preceding disclosure in any way whatsoever. One skilled in the art will recognize that the compounds represented by formula I can exist in one of two tautomeric forms, the enol or keto form. Those skilled in the art will promptly recognize appropriate variations from the procedures both as to reactants as well as to reaction conditions and techniques- Table II provides a summary of the EXAMPLE numbers, their chemical structures and their names.

CHART A REACTIONS PREPARATIONS FOR CHART A (oxynorbomene β-ketoamides)

Preparation Al. See CHART A, Step 1, Figure Al.

Step 1. The starting ketone, 1,3-Cyclopentenedione, and ftiran (used as both reactant and solvent) are refluxed for 3 to 5 days to produce the Diels-Alder adduct, Figure Al. Preparation A2. See CHART A, Step 2, Figure A2. EXAMPLE AXl. S-Hydroxy-S-oxo-lO-oxa-tricyclo .l.O^jdeca-S.S-diene^-carboxylic acid, 4-bromophenylamide.

4-BromophenyIisocyanate (7.24 g, 36.5 mmol) is dissolved in tetrahydrofuran (15 mL) and added to a solution of 5-Hydroxy-10-oxa-tricyclo[5.2.1.0 ^2,6 ]deca-4,8-dien-3-one (6.00 g, 36.5 mmol) in tetrahydrofuran (40 mL). Triethylamine (5.0 mL, 3-70 g, 36.5 mmol) is added and the reaction mixture stirred at room temperature for 3 hr. The reaction mixture is concentrated in vacuo and the residue taken up in EtOAc (50 mL) and washed with 2N H ₂ S0 ₄ (2 x 20 mL).

The organic phase is dried (Na^O.,) and concentrated in vacuo to a light, orange-brown powder. This is triturated with boiling MeOH (partial recrystallization). allowed to cool, filtered, and air dried to give 3-Hydroxy-5-oxo-10-oxa-tricyclo[5.2.1.0 ^2,6 ]deca-3.8-diene-4-carboxylic acid. 4-bromophenylamide as a light tan-colored crystalline solid. A further recrystallization from dilute MeOH gives a white crystalline solid: mp 180 °C; Η NMR (300 MHz, CDC1 ₃ ) δ 2.72 (d, J = 5.3 Hz, 1 H), 3.01 (d, J = 5.4 Hz, 1 H), 3.70 (broad s, 1H), 5.14 (s, 1 H), 6.54 (d, J = 1.8 Hz, 1 H), 7.48 (s, 4 H), 9.57 (s, 2 H). MS(EI) mle 361 (M ⁺ ), 363. IR (cm" ¹ , nujol): 1667, 1606. Anal. Calcd for C ₁₆ H ₁₂ BrN0 ₄ : C, 53.06; H, 3.34; N, 3.87. Found: C, 52.44; H, 3.23; N, 4.71. Following the general procedure of Preparation A2, above, except using the appropriate reactants, the following compounds are prepared.

EXAMPLE AX2. N-(3-Hydroxy-5-oxo-10-oxa- tricycIofS^.l.O^ _. dera-S.δ-diene^-carbony benzamide. MP 195°C. White solid from MeOH.

EXAMPLE AX3. 3-Hydroxy-5-oxo-10-oxa-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-caτboxylic acid, 4-trifluoromethylphenylamide MP 182°C. White crystals from MeOH.

EXAMPLE AX4. 3-Hydroxy-5-oxo-10-oxa-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxylic acid phenylamide. MP 148°. White solid from MeOH. EXAMPLE AX5.

3-Hydroxy-5-oxo-10-oxa-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxylic acid, 4-fluorophenylamide. MP 165°C. White solid from MeOH.

EXAMPLE AX6. 3-Hydroxy-5-oxo-10-oxa-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4 arboxylic acid, 4-chlorophenylamide. MP 180°C. White powder from MeOH.

EXAMPLE AX7. 3-Hydroxy-5-oxo-10-oxa-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxylic acid, 2,6-dichlorophenylamide. MP 191°C. White powder from MeOH.

EXAMPLE AX8. 3-Hydroxy-5-oxo- 10-oxa-tricyclo[5.2.1.0 ² ' ⁶ ]deca-3,8-diene-4-carboxylic acid, 3,5-dichlorophenylamide. MP 203°C. White powder from MeOH.

EXAMPLE AX9. 3-Hydroxy-5-oxo-10-oxa-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxylic acid, 2,6-difluorophenylamide. MP 195°C. White powder from MeOH. Preparation A3. See CHART A, Step 3, series a, page 1, Figure A3(a).

EXAMPLE AX11. δ^-Dibromo-S-hydroxy-S-oxo-lO-oxa-tricyclotS^.l.O^dec-S-ene ^-car-

boxylic acid, 4-bromophenylamide.

Bromine (49λ, 154 mg, 0.96 mmol) is added to a solution of 3-Hydroxy-5-oxo-10-oxa-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxylic acid. 4-bromophenylamide, Figure A2. (348 mg. 0.96 mmol) in CHC1 ₃ (25 mL). The reaction mixture is stirred overnight at room temperature and then concentrated in vacuo to a tan-colored solid. This is triturated with Et.0 to give the title compound, (mixture of cϊs-endo-, cis-exo- _> and trα/w-dibromoisomers) as an off-white powder- mp 212 °C; *H NMR (300 MHz, CDCI ₃ ) δ 2-81 (d, J = 6 Hz, 0.29 H), 2.88 (d, J = 5.8 Hz, 0.37 H), 3.11 (d, J = 6 Hz, 0.29 H), 3.18 (d, J = 5-9 Hz, 0.34 H), 3.57 (d. J = 5.8 Hz, 0.34 H), 3.91 (d, J = 5.8 Hz, 0.37 H), 3.98 (d, J = 3-1 Hz, 0-34 H), 4.01 (d, J = 3-1 Hz, 0.34 H), 4.35 (s, 0.58 H), 4.42 (m, 0.74 H), 4.78 (m, 0-68 H), 4-85 (m, 0.74 H), 4-90 (d, 0.58 H), 6.82 (broad s, 1H), 7.48 (s, 4H), 9.49 (s, 1H). MS(EI) mle 519 (M ⁺ ), 521, 523, 525. Anal- Calcd for C ₁₆ H ₁₂ Br ₃ N0 ₄ : C, 36.82; H, 2.32; N, 2.68. Found: C, 37-03; H, 2.35; N, 3.05.

Preparation A4- See CHART A, Step 4, series (a), page 2, Figure A4(a). EXAMPLE AX12. δ-Bromo-S-hydroxy-S-oxo-lO-oxa-tricyclofS^.l.C^^dec-S- ,8-diene-4-carboxyIic acid, 4-bromophenylamide

Potassium hydroxide (I mL, 45%) is added to a solution of 8,9-Dibromo-3-hydroxy-5~ oxo-10-oxa-tricyclo[5-2.1.0 ^2,6 ]dec-3-ene-4-carboxylic acid 4-bromophenylamide (60 mg, 0.12 mmol) and benzyltriethylammonium chloride (-7 mg) in EtOH (4 mL). The reaction mixture is stirred at room temperature for 10 days during which time the initially turbid reaction mixture becomes completely clear. The reaction mixture is concentrated in vacuo and the residue taken up in H ₂ 0 (5 mL). The strongly alkaline mixture is acidified with dilute H ₂ S0 ₄ to pH 1 and extracted with EtOAc. The organic phase is dried (NajSOJ and concentrated in vacuo to give the title compound as a pale-yellow powder. Η NMR (300 MHz, CDC1 ₃ , both keto-enol isomers are seen on the NMR time-scale) δ 2.85 (d, J = 5.4 Hz, 0.5 H), 2.90 (d, J = 5.4, 0.5 H), 3-15 (d, J = 5.3 Hz, 0.5 H), 3-19 (d, J = 5.4 Hz, 0.5 H), 4.96 (s, 1 H), 5.13 (s, 1 H), 6.54 (s, 0.5 H), 6.56 (s, 1 H), 7.47 (s, 4 H), 8.5 (broad s, 1 H), 9.51 (s, 1 H). MS(EI) mle 439 (M ⁺ ), 441, 443. Anal- Calcd for C ₁₆ H _n Br ₂ N0 ₄ : C, 43.57; H, 2.51; N, 3.18- Found: C, 43.31; H, 3.01; N, 3.00. Preparation A5. See CHART A, Step 3, series (b), page 3, Figure A3(b). EXAMPLE AX10. 3-bromo-N-(4-bromophenyl)-3a,4a,7,7a-8,8a-hexahydro-5-hydrox y-7-oxo- 4,8-Epoxy-4H-indeno[5,6-d]isoxazoIe-6-carboxamide,

A solution of AgN0 ₃ (281 mg, 1.66 mmol) in H ₂ 0 (1.5 mL) is added dropwise over a period of 60 min to a solution of 3-Hydroxy-5-oxo-10-oxa-tricyclo[5.2.1.0 ^2-6 ]deca-3,8-diene- -4-carboxylic acid, 4-bromophenylamide (Figure A2, EXAMPLE AXl) (300 mg, 0.828 mmol) and dibromofoπnaldoxime (336 mg, 1.66 mmol) in tetrahydrofuran (20 mL) at 25°C with vigorous stirring. The reaction mixture is stirred another 60 min and then diluted with EtOAc

(150 mL) and filtered. The filtrate is washed repeatedly with H ₂ 0 until the washings are no longer acidic. The organic phase is given a final wash with saturated brine and dried (N-i _j SOJ. Concentration in vacuo gives a solid which is triturated with boiling MeOH, cooled, filtered, and air-dried to give the title compound (282 mg, 70%) as an off-white, powdery solid consisting of a 1:1 mixture of exo-cis-exo (shown) and endo-cis-exo-iso τs: mp 162 °C; Η NMR (300 MHz, CDC1 ₃ ) δ 2.73 (d, J = 6 Hz, 0.5H), 2.81 (d, J = 6 Hz, 0.5 H), 3.01 (d. J = 6 Hz, 0.5 H). 3.10 (d, J = 6 Hz, 0.5 H), 3.70-3.74 (m, 1 H), 4.98 (m, 3 H), 5.50 (broad s, 2H), 7.48 (s, 4 H), 9.50 (broad s, 1 H). MS(EI) mle 482 (M ⁺ ), 484. IR (cm ^"1 , nujol): 1698. Anal. Calcd for C _I7 H ₁₂ Br ₂ N-0 ₅ : C, 42.18; H. 2.50; N, 5.79. Found: C, 42.02; H, 2.58; N, 5.93. Preparation A6. See CHART A, Step 3, series (c), page 4, Figure A3(c).

EXAMPLE AX13. 3-Hydroxy-5-oxo-10-oxa-tricyclo[5.2.1.0 ^2,6 ]dec-3-ene-4-carboxylic acid. 4-bromophenylamide.

Ten percent palladium on charcoal (100 mg) is added to a solution of 3-Hydroxy-5-oxo-10-oxa-tricyclo[5.2.1.0 ² ' ⁶ ]deca-3,8-diene-4-carboxylic acid 4-bromophenylamide, Figure A2, (2.00 g, 5.52 mmol) in tetrahydrofuran (200 mL) and the mixture shaken for 60 min with H ₂ (40 psia). The reaction mixture is flushed with N ₂ , filtered through celite, and the filtrate concentrated in vacuo. The residue is triturated with boiling MeOH, cooled, filtered, and air-dried to give the title compound as a tan-colored, crystalline solid: mp 193 °C; ^] H NMR (300 MHz, CDC1 ₃ ) δ 1.63 (m, 2 H), 1.87 (m, 2 H), 2.71 (d, J = 5.67 Hz, 1 H), 3.00 (d, J = 5.69 Hz, IH), 4.75 (m, 2 H), 6.77 (broad s, 2 H), 7.46 (m, 4H), 9.66 (broad s, 1 H). MS(EI) mle 363 (M ⁺ ), 365. Anal. Calcd for C ₁₆ H ₁₄ BrN0 ₄ : C, 52.77; H, 3.88; N, 3.85. Found: C, 52.30; H, 3.63; N, 4.03.

Preparation A7. See CHART A, Step 3, series (d), page 5, Figure A3(d). EXAMPLE AX14. 3-Methoxy-5-oxo-10-oxa-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxylic acid, 4-bromophenylamide.

A solution of diazomethane (CH ₂ N ₂ ) in Et-,0 (0.5 M, 80 mL, 40 mmol) is prepared as follows: l-methyl-3-nitro-l-nitrosoguanidine (MNNG) (6.00 g, 40.8 mmol) is added in several small portions with intervening swirling to a two-phase solution of KOH (24 mL, 45%) and E O (80 mL) maintained near 0°C in a scratch-free, rubber-stoppered, 250 mL Erienmeyer flask. When all the MNNG is added, the two-phase reaction mixture is intermittently swirled for two minutes. The upper-layer containing the diazomethane is dried by carefully decanting it into another scratch-free, rubber-stoppered, 250 mL, Erienmeyer flask containing several KOH pellets maintained near 0°C. The flask is kept near 0°C and intermittently swirled for two minutes after which the diazomethane solution is either used or briefly stored at 0°C in a stoppered flask. A portion of the diazomethane solution (40 mL, 0.5 M, 20 mmol) is added all at once to a solution of 3-Hydroxy-5-oxo-10-oxa-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxylic acid,

4-bromophenylamide, Figure A2, (1.00 g, 2.76 mmol) in tetrahydrofuran (100 mL) at room temperature. The reaction mixture is allowed to stand 6 minutes. The reaction is quenched by adding glacial HO Ac (3 mL) causing instantaneous discharge of the yellow color. The reaction mixture is concentrated in vacuo and the residue taken up in EtOAc (50 mL). The resulting solution is washed with 1 N NaOH (2 x 50 mL), samrated NaCl (20 mL), dried (Na-S0 ₄ ). and concentrated in vacuo to a clear, yellow oil containing some solid. This is taken up in boiling MeOH, cooled, and filtered to give the title compound as an off-white, crystalline solid: mp 159 °C; Η NMR (300 MHz, DMSO-D ₆ ) δ 2.73 (d, J = 5.5 Hz, 1 H). 3.24 (d, J = 5.5 Hz. 1 H). 4.32 (s, 3 H), 5.10 (s, 1 H). 5.12 (s. 1 H), 6.52 (dd, J = 1.3 Hz, 5-6 Hz, 1 H), 6.58 (dd, J = 1-3 Hz, 5.8 Hz, 1 H), 7.44 (m. 4 H), 10.3 (s, 1 H). MS(EI) mle 375 (M ⁺ ), 377. Anal. Calcd for C ₁₆ H _n F ₂ N0 ₄ : C, 54.28; H, 3.75; N, 3.75. Found: C, 53.21; H, 3.65; N, 4.05.

Preparation A8- See CHART A, Step 4, series (d), page 5, Figure A4(d). EXAMPLE AX15. 3-Amino-5-oxo-10-oxa-tricyclo[5.2.1.0 ² ' ⁶ ]deca-3,8-diene-4-carboxylic acid. 4-bromophenylamide. Anhydrous ammonia is slowly bubbled into a slurry of

3-metJιoxy-5-oxo-10-oxa-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxylic acid, 4-bromo-phenyI- amide, Figure A3(d), (300 mg, 0.797 mmol) in MeOH (15 mL) at room temperature during 15 min. The temperature of the reaction mixture rose to 40°C and then subsided. Stirring, is con¬ tinued for another 30 min. and the reaction mixture concentrated in vacuo to a solid. This is triturated with boiling MeOH, chilled, filtered, and air-dried to give the title compound as a white powder mp 238 °C; 'H NMR (300 MHz, CDC1 ₃ ) δ 2.35 (m, 1 H), 2.66 (d, J = 5.8 Hz, I H), 4.81 (s, 2 H), 6.22 (d. J = 5.8 Hz, 1 H), 6.26 (d, J = 5.8 Hz, 1 H). 7.13 (d, J = 8.4 Hz, 2 H), 7.27 (d, J = 8.4 Hz, 2 H). 8.56 (m, 2 H ), 10.2 (s, 1 H). MS(EI) mle 360 (M ⁺ ), 362. Anal. Calcd for C ₁₆ H _I3 BrN ₂ 0 ₃ : C, 53.21; H, 3.63; N, 7.76. Found: C, 51.89; H, 3.58; N, 7.64. CHART B REACTIONS

PREPARATIONS FOR CHART B (norbomene β-ketoamides) Preparation Bl. See CHART B, Step 1, Figure Bl. EXAMPLE BX1 - not claimed.

Freshly distilled cyclopentadiene and maleic anhydride ^ tlS' ) undergo a Diels- Alder reaction to produce the compound shown in Figure Bl. Preparation B2 See CHART B, Step 2, Figure B2. EXAMPLE BX2. 3-Ethoxycarbonylacetyl-bicyclo[2.2.1]hept-5-ene-2-carboxylic acid.

A solution of n-butyllithium in hexane (190 mL, 1.6 M, 305 mmol) is added dropwise to a solution of diisopropylamine (43 mL, 30.8 g, 305 mmol) in dry tetrahydrofuran (200 mL) at 0°C. The reaction mixture is stirred for 15 min, cooled to -78°C, and a solution of EtOAc (29.8 mL, 26.8 g, 305 mmol) in dry tetrahydrofuran (100 mL) added dropwise. After stirring 15 min,

a solution of EXAMPLE BX1, (25.0 g, 152 mmol) in dry tetrahydrofuran (400 mL) is added dropwise. The reaction mixture is stirred for 2 hrs at -78 ^C C, the reaction is then quenched by adding glacial HOAc (38 mL) all at once, and the mixture allowed to warm to room temperature. Concentration in vacuo gave a milky oil. This is taken up in water (800 mL), and crushed ice added to reduce the temperature to <10°C. The mixture is acidified to pH 1 by adding 6N H ₂ S0 ₄ and then extracted with EtOAc (2 x 400 mL). The extracts are combined, dried (Na ₂ S0 ₄ ), and concentrated in vacuo. Residual HOAc is removed azeotropically by in vacuo concentration from toluene (2 x 200 mL) to give a light-yellow, turbid oil. This is crystallized from toluene (100 mL) by allowing the solution to stand overnight at room temperature. The crystals are washed with hexane and air-dried to give the title compoimd, a mixture of diastereomers, as large, white, dense crystals: mp 91 °C; Η NMR (300 MHz, CDC1 ₃ ) δ 1.30 (m, 3 H), 1.41 (d, J = 8.5 Hz, 0.5 H), 1.47 (d, J = 8.5 Hz, 0.5 H). 1.58 (d, J = 8.1 Hz, 0.5 H), 1.67 (d, J = 8.5 Hz, 0.5 H), 2.76 (q, J = 16.5 Hz, 2 H), ^" 3.04 (m, 1 H), 3.11 (s, 0.5 H), 3.18 (s, 0.5 H), 3.28 (s, 1 H), 3.45 (m, 0.5 H), 3.54 (m, 0.5 H), 4.21 (m, 2 H), 6.22 (m, 2 H). MS(EI) mle 252 (M ⁺ ). Anal. Calcd for C ₁₃ H ₁₆ 0 ₅ : C, 61.90; H, 6.39; N, 0.00. Found: C, 61.63; H, 6.53; N, 0.19.

Preparation B3. See CHART B, Step 3, page 2, Figure B3. EXAMPLE BX4. 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxylic acid, ethyl ester. Powdered carbonyldiimidazole (643 mg, 3.96 mmol) is added in a single portion to a solution of 3-Ethoxycarbonylacetyl-bicyclo[2.2.1]hept-5-ene-2-carboxylic acid. EXAMPLE BX2, (l.OOg, 3.96 mmol) in tetrahydrofuran (20 mL) and the mixture heated under reflux. In two hours evolution of C0 ₂ stops, and the reaction mixture is cooled to 0°C. A solution of potassium tert-butoxide in tetrahydrofuran (1.0 M, 7.9 mL, 7.93 mmol) is added dropwise. Stirring is continued at 0°C for 30 minutes and then overnight at room temperature. The mix¬ ture is concentrated in vacuo and the residue taken up in 2N H ₂ S0 ₄ (50 mL). This is extracted with EtOAc (2 x 30 mL), and the extracts combined, dried (Na ₃ S0 ₄ ), and concentrated in vacuo to a clear, light-yellow oil. This is triturated in a small amount of n-hexane at room temperature, filtered, and air-dried to give the title compoimd as a white powder: mp 133 °C; Η NMR (300 MHz, CDC1 ₃ ) δ 1.37 (t, J = 7.2 Hz, 3 H), 1.58 (d, J = 8.6 Hz, 1 H), 1.78 (d, J = 8.6 Hz, 1 H), 3.19 (broad s, 2 H), 3.27 (s, 2 H), 4.35 (q, J = 7.2 Hz, 2 H), 6.01 (s, 2 H). ^I3 C NMR (75 MHz, CDC1 ₃ ) δ 13.8, 43.9, 52.1, 61.1, 109.5, 133 (broad), 167.7, 198 (broad). MS (EI) mle 234 (M ⁺ ). Anal. Calcd for C ₁₃ H ₁₄ 0 ₄ : C, 66.66; H, 6.03; N, 0.00. Found: C, 66.32; H, 5.94; N, 0.38. Preparation B4. See CHART B, Step 4, page 4, Figure B4.

EXAMPLE BX5. S-Hydroxy-S-oxo-tricyclofS^.l.O^jdeca-S.S-diene^-cartoxylic acid,

4-bromophenylamide.

3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ^2,6 ] deca-3.8-diene-4-carboxylic acid, ethyl ester, EXAMPLE BX4, Figure B3, (213 mg. 0.909 mmol) and 4-bromoaniline (156 mg. 0.909 mmol) are combined with toluene (5 mL). and the mixture heated under reflux for 90 minutes. The reaction mixture is concentrated in vacuo and the residue crystallized from n-hexane to give the title compound (134 mg, 41%) as a white crystalline solid: -R NMR (300 MHz, CDCI ₃ ) δ 1.63 (d, J = 8.5 Hz, 1 H), 1.84 (d, J = 8.7 Hz, 1 H), 3.12 (dd, J = 6.13 Hz, 4.50 Hz, 1 H), 3.28 (m, 2 H), 3.40 (dd, J = 6.24 Hz, 4.39 Hz, 1 H), 5.98 (s, IH), 6.04 (s, IH), 7.46 (m, 4 H). MS (EI) mle 359 (M ⁺ ), 361. Anal. Calcd for C _π H, ₄ BrN0 ₃ : C, 56.69; H, 3.92; N, 3.89. Found: C, 56-42; H, 3-76; N, 3.69.

Following the general procedure of Preparation B4, above, except using the appropriate reactants, the following compounds are prepared.

EXAMPLE BX6. S-Hydroxy-S-oxo-tricyclo^.l.O^deca-S^-diene-^-carboxylic acid, 5-trifluoromethyI-[l,3,4]thiadiazol-2-ylamide. MP 156°C. Tan-colored powder from EtOAc- EXAMPLE BX7. 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ² ' ⁶ ]deca-3,8-diene-4-carboxylic acid,

4-trifluoromethylphenylamide. MP 171°C. White crystals from EtOAc/hexane.

EXAMPLE BX8. 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ² - ⁶ ]deca-3,8-diene- -carboxyIic acid, 4-cyanophenylamide. MP 216°C. White crystals from EtOAc.

EXAMPLE BX9. 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ² - ⁶ ]deca-3,8-diene-4-carboxylic acid, 4-nitrophenylamide. MP 206°C. Off-white crystals from EtOAc.

EXAMPLE BX10. S-Hydroxy-S-oxo-tricyclo^^-LO^Jdeca-S^-diene^-carboxylic acid, 4-methoxyphenyIamide. MP 138°C. Light-brown crystals from EtOAc/hexane.

EXAMPLE BX11. 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 2,6]deca-3,8-diene-4-carboxylic acid. 2-methyl-4.-(4-trifluoromethylphenoxy)phenylamide. MP 165°C. White powder from MeOH.

EXAMPLE BX12. 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxylic acid, 3.5-bis-trifluoromethylphenylamide. MP 159°C- Pale-yellow crystals from MeOH.

EXAMPLE BX13. S-Hydroxy-S-oxo-tricyclot^-l.O^deca^δ-diene^-carboxylic acid, 2-fluorophenylamide. MP 131°C. White powder from MeOH. EXAMPLE BX14. 3-Hydroxy-5-oxo-tricyclo[5-2.1.0 ^2,6 ]deca-3,8-diene- -carboxylic acid,

3-fIuorophenylamide. MP 128°C. White crystals (plates) from MeOH.

EXAMPLE BX15. 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ² ' ⁶ }deca-3,8-diene-4-carboxylic acid, 2-trifluoromethyIphenyIamide. MP 92°C. Pale-yellow powder from MeOH.

EXAMPLE BX16. S-Hydroxy-S-oxo-tricyclo ^.l.O^deca^δ-diene^-carboxylic acid, 2,6-difluorophenylamide. MP 177°C. White crystals from MeOH.

EXAMPLE BX17. 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ² ' ⁶ ]deca-3,8-diene- -carboxyIic acid,

2,6-difluorobenzylamide. MP 113°C. Tan-colored powder from MeOH.

EXAMPLE BX18. 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ^2,6 ]deca-3.8-diene-4-caiPoxylic acid 2,4,6-trifluorophenylamide. MP 190°C. White granular solid from MeOH/hexane.

EXAMPLE BX19. 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxylic acid 3,5-difluorophenylamide. MP 160°C. Pale-yellow powder from MeOH trituration.

EXAMPLE BX20. 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxyiic acid 4-fluorophenyl amide. MP 131°C. White crystals from MeOH.

EXAMPLE BX21. S-Hydroxy-S-oxo-tricyclo^.l.O^Jdeca-S^-diene^-carboxylic acid 9H-fluoren-2-ylamide. MP 238°C. Light-brown powder from MeOH trituration. EXAMPLE BX22. 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxyIic acid

7-bromo-9H-fluoren-2-ylamide. MP 262°C. Cream-colored powder from MeOH trituration.

EXAMPLE BX23. S-Hydroxy-S-oxo-tricyclofS^.l.O^jdeca-S.S-diene^-carboxylic acid, 9-oxo-9H-fluoren-2-ylamide. MP 230°C. Green-brown powder from MeOH trituration.

EXAMPLE BX24. 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxylic acid biphenyl-4-ylamide. MP 174°C. White powder from MeOH trituration.

EXAMPLE BX25. S-Hydroxy-S-oxo-tricyclo^^.l^^deca-S.δ-diene^-carboxylic acid 2,2'-dimethylbiphenyl-4-ylamide. Tan-colored powder from MeOH.

Following the general procedure of Preparation B4, above, plus preparation A3 from the CHART A reactions, except using the appropriate reactants, the following compound, Example BX26, is prepared.

EXAMPLE BX26. 2-(N-(4-trifluoromethylphenyl)carbamoyl)-5H,6H-dibromo-4H,7H - methano-3-hydroxyinden-l-one. MP 221°C. Cream-colored powder from hexane trituration.

Following the general procedure of Preparation B4, above, plus preparation A6 from the CHART A reactions, except using the appropriate reactants, the following compounds, examples BX27 and BX28 are prepared.

EXAMPLE BX27. S-Hydroxy-S-oxo-tricyclofS^.l.O^jdec-S-ene^-carboxylic acid, 4-bromophenylamide. MP 156°C. Light-brown crystals from MeOH.

EXAMPLE BX28. 3-Hydroxy-5-oxo-tricyclo[5.2.1.0 ² - ⁶ ]deca-3,8-diene* -carboxyIic acid, 4-aminophenylamide. MP 189°C. Pale-yellow solid from EtOAc trituration. Following the general procedure of Preparation B4, above, plus preparation A7 from the

CHART A reactions, except using the appropriate reactants, the following compounds, examples BX29 and BX30, are prepared.

EXAMPLE BX29. 3-Methoxy-5-oxo-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxyIic acid ethyl ester. White crystals from pale-yellow oil. EXAMPLE BX30. 3-Methoxy-5-oxo-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxyUc acid biphenyl-4-ylamide. MP 201°C. White crystals from MeOH.

Following the general procedure of Preparation B4, above, plus preparation A8 from the CHART A reactions, except using the appropriate reactants, the following compounds, examples BX31 and BX32, are prepared.

EXAMPLE BX31. S-Methoxy-S-oxo-tricyclofS^.l.C ^deca-S.S-diene^-carboxylic acid, 4-trifluoromethylphenylamide. MP 190°C. White crystals from MeOH.

EXAMPLE BX32. S-Amino-S-oxo-tricyclofS^-l.O^^deca-S.S-diene-^carboxylic acid, 4-trifluoromethylphenylamide. White crystals from MeOH.

CHART C REACTIONS PREPARATIONS FOR CHART C (tetracyclic β-ketoamides) Preparation C See CHART C, Step 1, Figure Cl.

Cyclooctatetraene (8.73 g, 83.8 mmol) and maleic anhydride (8.22 g, 83.8 mmol) are combined with toluene and heated under reflux for 24 hrs. The reaction mixmre is concentrated in vacuo to a cream-colored solid. This is recrystallized from 1:2 EtOAc:hexane to give the Diels-Alder adduct an anhydride, as a white crystalline solid: ^l R NMR (300 MHz, CDC1 ₃ ) δ 2.81 (s, 2 H), 3.08 (s, 2 H), 3.24 (s, 2 H), 5.92 (s, 2 H), 6.04 (m, 2 H). Preparation C2. See CHART C, Step 2, Figure C2.

The adduct, anhydride from preparation Cl, above, is reacted with ethyl lithioacetate in tetrahydrofuran at -78°C to give the β-ketoester acid. Figure C2. Preparation C3. See Chart C, Step 3, Figure C3. The β-ketoester acid, Figure C2, is converted to the ester, Figure C3, by heating with carbonyl diimidazole and potassium tert-butoxide in tetrahydrofuran at reflux. Preparation C4. See Chart C, Step 4, Figure C4.

EXAMPLE CXI. (2a.alpha., 3-beta., 3a-beta., 6a.beta., 7-beta., 7a-alpha.)-N-(4- bromophenyl)-2a,3a,4,6a,7,7a-hexahydro-6-hydrOxy-4-oxo-3,7-E theno-3H-cycIobut[f]indene-5- carboxamide, MP 232°C. Pale yellow powder from reaction mixmre.

The ester, Figure C3, from above, is heated with 4-bromoaniIine in toluene at reflux, according to the general procedures provided in preparation B4 from the reactions of CHART B, except using the appropriate reactants, to give the title compound-

Following the general procedure of Preparation C4, above, plus the procedures provided in preparation B4 from the reactions of CHART B, except using the appropriate reactants, the following compounds, examples CX2 through CX6, are prepared-

EXAMPLE CX2. (2a.alpha., 3.beta., 3a-beta-, 6a.beta., 7.beta., 7a.alpha.)-N-(5- trifluoromethyi-[l,3,4]thiadiazol-2-yl)-2a,3a,4,6a,7,7a-hexa hydro-6-hydroxy-4-oxo-3,7-Etheno- 3H-cycIobut[fIindene-5-carboxamide, MP 235°C. White powder from EtOAc. EXAMPLE CX3. (2a.alpha., 3.bet&, 3a-beta., 6a.beta., 7.beta., 7aalpha.)-N-[4-

(trifluoromethyI)phenyI]-2a,3a,4,6a,7,7a-hexahydro-6-hydr oxy-4-oxo-3,7-Etheno-3H-

cyclobut[f]indene-5-carboxamide, MP 197°C. Off-white powder from EtOAc & hexane.

EXAMPLE CX4. (2a.alpha.. 3.beta., 3a.beta., 6a.beta., 7.beta., 7a.alpha.)-N-(5- bromopyridin-2-yl)-2a,3a,4,6a,7,7a-hexahydro-6-hydroxy-4-oxo -3,7-Etheno-3H- cyclobut[f]indene-5-carboxamide, MP 239°C. Off-white powder from trituration with hot EtOAc.

EXAMPLE CX5. (2a.alpha., 3.beta., 3a.beta., 6a.beta., 7.beta., 7a.alpha.)-N-(4- cyanophenyl)-2a,3a,4,6a,7,7a-hexahydro-6-hydroxy-4-oxo-3,7-E theno-3H-cyclobut[f]indene-5- carboxamide, MP 253°C. Off-white powder from trituration with hot EtOAc.

EXAMPLE CX6. (2a.alpha., 3.beta., 3a.beta., 6a-beta., 7.beta., 7a.alpha.)-N-[[2-methyI- 4-(4-trifluoromethyl)phenoxy]phenyl]-2a,3a,4,6a,7,7a-hexahyd ro-6-hydroxy-4-oxo-3,7-etheno-3H- cyclobut[f]indene-5-carboxamide, MP 167°C. White powder from trituration with hot MeOH.

CHART D PREPARATIONS FOR CHART D (fulvene β-ketoamides) Preparation PL See CHART D, Step 1, Figure DI. A methanol solution of freshly distilled cyclopentadiene with acetone is stirred in the presence of pyrrolidine to produce 6,6-dimethylfulvene, Figure DI.

Preparations D2 through D5 are "series (a)" preparations. Preparation D2. See CHART D, Step 2, series (a), Figure D2(a). The fulvene from preparation DI, above, is refluxed with maleic anhydride in ether overnight to produce the anhydride, Figure D2(a).

Preparation D3. See CHART D, Step 3, series (a). Figure D3(a). The anhydride from preparation D2, above, Figure D2(a), is treated with ethyl lithioacetate at -78°C to give the β-ketoester acid as a yellow oil containing impurities, Figure D3(a). Preparation D4. See CHART D, Step 4, series (a), Figure D4(a).

Without purification the β-ketoester acid, Figure D3(a), from preparation D3, above, is converted to a cyclized ester by heating it with carbonyldϋmidazole and potassium tert-butoxide in tetrahydrofuran, the temperature should not be allowed to exceed 40°C. The cyclized ester, produced as a dark red oil, is shown in Figure D4(a). Preparation D5. See CHART D, Step 5, series (a), Figure D5(a).

The cyclized ester from Preparation D4, Figure D4(a) is then used following the general procedure of Preparation B4, from the reactions of CHART B, except using the appropriate reactants, to prepare examples DX1, DX2 and DX3.

EXAMPLE DX1, 3-Hydroxy-lO-isopropylidene -5-oxo-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxylic acid, 4-bromophenylamide. Light tan- colored crystals from MeOH.

EXAMPLE DX2. 3-Hydroxy-10-isopropylidene -5-oxo-tricyclo[5.2. l.O^Jdeca-S.S-diene-^carboxylic acid, 4-trifluoromethylphenylamide. Crystalline solid from MeOH.

EXAMPLE DX3. 3-Hydroxy-10-isopropylidene- 5-oxo-tricyclo[5.2.1.0 ^2,6 ]deca-3,8-diene-4-carboxylic acid, 4-cyanophenylamide. Pale-yellow powder from MeOH.

Preparations D6 through D7 are "series (b)" preparations. Preparation D6. See CHART D, Step 2, Series (b). Figure D2(b). 6,6-DimethyIfulvene (5.00g, 47.1 mmol) is added to a solution of cyclopentenedione (2.26 g, 23.5 mmol) in Et0 ₂ (20 mL). The mixture is stirred for 6 days at room temperamre. chilled to 0°C for two hrs and then filtered. The solids are washed with n-hexane to give Hydroxy-lO-isopropylidene-tricyclotS^.l.O^deca-^.S-dien-S-on e, 1:1 mixture of exo and endo isomers, as a white powdery solid: mp 158 °C; Η NMR (300 MHz, CDC1 ₃ ) δ 1.52 (s, 3 H), 1.56 (s, 3 H ), 2.52 (s, 1 H), 2.98 (m, 1 H), 3.46 (m, 1 H), 3.59 (m, 1 H), 5.16 (s, 0.5 H), 5.21 (s, 0.5 H), 6.10 (t, J = 2.0 Hz, 1 H), 6.37 (t J = 1.9 Hz, 1 H).

Preparation D7. See CHART D, Step 3, Series b, Figure D3(b). The compound from Preparation D6, Figure D2(b) is then used following the general procedure of Preparation A2, from the reactions of CHART A, except using the appropriate reactants, to prepare examples DX4, and DX5. EXAMPLE DX4. 3-Hydroxy-10-isopropylidene-

5-oxo-tricycIo[5.2.1.0 ^2, ^deca-3,8-diene-4-carboxylic acid. 4-bromophenylamide, 1:1 mixmre of exo and endo isomers. MP 159°C. Off-white powder from MeOH. Light tan-colored crystals from MeOH.

EXAMPLE DX5. N-(3-Hydroxy-5-oxo- tricycIo[5.2.1.0"]deca-3,8-diene-4-carbonyI)-benzamide. MP 66°C. Tan-colored solid foam.

CHART A Page 1

Step 1

(Al)

Step 2

CHART A

Series (a) page 2

Step 3(a)

Step 4(a)

CA4(a))

CHART A Series (b) Page

Step 3(b)

CHART A

Series (c)

Page 4

Step 3(c)

CHART A

Series (d)

Page 5

Step 3(d)

(A3(d))

Step 4(d)

CHART B Page 1

Step 1

Step 2

Step 3

CHART B Page 2

Step 3

Step A

-

(B4)

CHART C Page 1

Step 1

Step 2

0 0

Step 3

CHART C Page 2

Step 3

(C3)

Step 4

(C4)

CHART D

Series (a) & (b)

Page 1

Step 1

(02(a)) (D2(b))

Step 3 (b) Series (b)

(D3(b))

CHART D

Series (a)

Page 2

Step 3 Series (a)

Step 4(a)

(D4(a))

Step 5(a)

(D5(a))

TABLE I - Biological Data

TABLE II - Compounds

TABLE π - Compounds (continued)

'ϊϊβ?--:-:- r r. 'r _rτ

TABLE II - Compounds (continued)

TABLE II - Compounds (continued)

TABLE II - Compounds (continued)

SUBSTITUTE SHEET

TABLE II - Compounds (continued)

SUBSTITUTE SHEET

TABLE π - Compounds (continued)

SUBSiliu

χbiύ glli-fiL. SHEET

ύύtiύillϋli SUB

TABLE II - Compounds (continued)

SHEET

β.ι χ.z>τ?_- <_τ ITT^ FT

TABLE π - Compounds (continued)

Compound Compound Structure Compound Name No.

BX30 3-Methoxy-5-oxo-tricyclo[5.2.1.0 ^2,6 ]d eca-3,8-diene-4-caιboxylic acid, biphenyl-4-ylamide

BX31 S-Methoxy-S-oxo-tricycIotS^.l. ^d eca-3,8-diene-4-carboxyIϊc acid, 4-trifluoromethylphenylamide

BX32 3-Amino-5-oxo-tricyclo[5.2. LO^ dec a-3,8-diene-4-carboxylic acid, 4-trifluoromethylphenylamide

CXI (2a.alpha., 3.beta., 3a.beta., 6a.beta.,

7.beta., 7aalpha.)-

N-4 ^l -bromophenyl)-2a,3a,4,6a,7,7a- hexahydro-6-hydroxy-4-oxo-,

3,7-Etheno-3H-cyclobut[f]indene-5- carboxamide,

SUBSTITUTE SH £--. !__-_

TE SHEET

TABLE π - Compounds (continued)

15

SUBSTITUTE SHEET

SUBSTITUTE SHEET

SUBST-TUTE SHEET