BACKGROUND OF THE INVENTION Field of the Invention The present invention relates generally to the fields of organic chemistry and novel inhibitors of cell proliferation. More specifically, the present invention relates to novel p- heteroatom-substituted phenols and analogs thereof, and their use as anti-proliferative agents. Description of the Related Art

The control of cell proliferation is a complex process that involves multiple interacting components. Whether a cell grows or not depends on the balance of the expression of negatively-acting and positively-acting growth regulatory genes. Negatively-acting growth regulatory genes are those that, when expressed in or provided to a cell, lead to suppression of cell growth. Positively-acting growth regulatory genes are those which, when expressed in or provided to a cell, stimulate its proliferation. Recently, several negatively-acting growth regulatory genes, called tumor suppressor genes, which have a negative effect on cell proliferation have been identified. These genes include, but are not limited to, the human retinoblasto a gene, RB-1, and the p53 gene. The absence or inactivation of some of these negative growth regulatory genes has been correlated with certain types of cancer.

There is a wide variety of pathological cell proliferative conditions for which novel methods are needed to provide therapeutic benefits. These pathological conditions may occur in almost all cell types capable of abnormal cell proliferation. Among the cell types that exhibit pathological or abnormal growth are (1) fibroblasts, (2) vascular endothelial cells, and (3) epithelial cells. It can be seen from the above that methods are needed to treat local or disseminated

pathological conditions in all or almost all organ and tissue systems of the individual.

Melanoma, the most virulent of skin cancers, is a highly metastatic disease affecting both sexes and is almost uniformly fatal within five years of diagnosis. Surgical removal of localized malignancies has proven effective only when the disease has not spread beyond the primary lesion. Once the disease has spread, the surgical procedures must be supplemented with other more general procedures to eradicate the diseased or malignant cells. Most of the commonly utilized supplementary procedures such as irradiation or chemotherapy are not localized to the tumor cells and, although they have a proportionally greater destructive effect on malignant cells, often affect normal cells to some extent. Sterically hindered phenols are an important class of peroxyl trapping antioxidants in both biological and non- biological environments. For example, α-tocopherol, a component of vitamin E, is the major lipid soluble antioxidant in human blood, and its biological function, as well as that of its analogs, continues to be of considerable interest. The structural features responsible for the high antioxidant activity of α-tocopherol and its analogs are known, at least as measured in homogeneous solution, and numerous papers discuss determination of rate constants, k _1f for the chain-terminating reaction of phenols with peroxyl radicals (eq. 1) . Complementary studies have been reported in miceliar systems and more recently in environments designed to model biological membranes.

ROO- ₊ ArOH →- OOH + ArO- (1)

The relative efficacies of the phenolic antioxidants, as reflected in k.,, as well as the absolute values of these rate constants, are media dependent. The major factor defining the free radical chain-terminating activity of the phenols is the

nature of the substitution on the aromatic ring. Ortho-, meta-, and para-alkyl, para-alkoxy, and para-alkylthio groups are seen to augment the reactivity of the phenols toward radicals, owing to stabilization of the incipient phenoxy1 radical character of the transition state for hydrogen atom transfer. Epr studies have confirmed that the heteroato -containing substituents interact with the unpaired spin in the phenoxyls. Moreover, stereo-electronic effects appear to be important in the case of alkoxy and alkylthio groups. The prior art is deficient in the lack of effective means of inhibiting the undesirable or uncontrollable cell proliferation in a wide variety of pathophysiological conditions. The present invention fulfills this long-standing need and desire in the art. SUMMARY OF THE INVENTION

In one embodiment of the present invention, there are provided antiproliferative p-heteroatom-substituted phenolic compounds, or derivatives thereof, having a structural formula selected from the group consisting of

wherein m is 1 to 3, n is 0 when Het is oxygen or sulfur, 0 to 4 when Het is nitrogen, Het is nitrogen, oxygen or sulfur; wherein R is selected from the group consisting of hydrogen, alkyl, arymethyl and acyl; R ¹ is alkyl; R ² is selected from the group consisting of hydrogen and alkyl; R ³ is selected from the group consisting of alkyl and acyl when Het is nitrogen, : (electron pair) when Het is oxygen or sulfur; R ⁴ is selected from the group consisting of hydrogen and alkyl; R ⁵ is selected from

the group consisting of hydrogen and alkyl; and R ⁶ and R ⁷ are selected from the group consisting of hydrogen, alkyl and oxygen.

In another embodiment of the present invention, there is provided a pharmaceutical composition comprising the compound of claim 1 and a pharmaceutically acceptable carrier.

In yet another embodiment of the present invention, there is provided a method for the treatment of a pathological cell proliferative disease comprising administering to an animal a pharmacologically and therapeutically effective dose of a pharmaceutical composition comprising a p-heteroatom-substituted phenol or an analog thereof.

Other and further aspects, features, and advantages of the present invention will be apparent from the following description of the presently preferred embodiments of the invention given for the purpose of disclosure.

BRIEF DESCRIPTION OF THE DRAWINGS

So that the matter in which the above-recited features, advantages and objects of the invention, as well as others which will become clear, are attained and can be understood in detail, more particular descriptions of the invention briefly summarized above may be had by reference to certain embodiments thereof which are illustrated in the appended drawings. These drawings form a part of the specification. It is to be noted, however, that the appended drawings illustrate preferred embodiments of the invention and therefore are not to be considered limiting in their scope.

Figure 1 shows the structures of the antiproliferative p-heteroatom-substituted phenolic compounds, or derivatives thereof, of the present invention. Figure 2 shows individual structures and compound names for selected antiproliferative p-heteroatom-substituted phenol, or derivatives thereof, compounds of the present invention.

Figure 3 shows the synthetic scheme for the synthesis of 9-acetoxy-8,10-dimethyljulolidine (6).

Figure 4 shows the synthetic scheme for the synthesis, among others, of W-isobutyryl-5-hydroxy-3,4,6-trimethylindoline

(11b), W-pivaloyl-5-hydroxy-3,4,6-trimethylindoline (lie), N- isobutyryl-5-acetoxy-3,4,6-trimethylindoline (12b), and N- isobutyryl-5-methoxy-3,4,6-trimethylindoline (13) .

Figure 5 shows the synthetic scheme for the synthesis of lV-acetyl-5-hydroxy-3,3,4,6-tetramethylindoline (16).

Figure 6 shows the synthetic scheme for the synthesis of W-pivaloyl-5-hydroxy-3,3,4,6-tetramethylindoline (20).

Figure 7 shows the synthetic scheme for the synthesis of 4,6-di-tert-butyl-2,3-dihydro-5-hydroxy-2 ( 3H)-benzofuran one (24).

Figure 8 shows the synthetic scheme for the synthesis of iV-pivaloyl-5-hydroxy-3,4,6,7-tetramethylindole (27).

Figure 9 shows the synthetic scheme for the synthesis of N-acetyl-5-hydroxy-3,4,6,7-tetramethylindoline (29b) Figure 10 shows the synthetic scheme for the synthesis of N-acetyl-5-hydroxy-3,3,4,6,7-pentamethylindoline (31b) DETAILED DESCRIPTION OF THE INVENTION Thepresent inventionprovidesp-heteroatom-substituted phenols and derivatives thereof in the treatment of cancers and disorders involving excess cell proliferation. The general structures of the novel compounds of the present invention are shown in Figure l and possible routes for their syntheses are provided in Figures 2-10. These molecules are in the class of compounds known as phenols or simple derivatives thereof. ' The therapeutic use of the compounds of the present invention in treatment of cancers and other diseases and disorders involving excess cell proliferation is illustrated. The 5-hydroxyindoline derivatives were shown at various concentrations to inhibit the proliferation of human breast cancer cells (MDA MB 435 and MCF-7 breast cancer cells) and human pro yelocytic leukemia cells (HL-60) ) leukemia cells) . The novel compounds of the present invention are strong inhibitors of cell growth. The novel features of these molecules include, inter alia, the nature of substituents on the phenolic ring and the fusion of a heteroatom-containing ring to it.

The novel compounds and methods of present invention may be used to treat either neoplastic diseases and non-

neoplastic diseases. Representative examples of neoplastic diseases are ovarian cancer, bladder cancer, lung cancer, cervical cancer, breast cancer, prostate cancer, gliomas, fibrosarcomas, retinoblastomas, melanomas, soft tissue sarcomas, osteosarcomas, colon cancer, carcinoma of the kidney and pancreatic cancer.

Representative examples of non-neoplastic diseases are selected from the group consisting of psoriasis, benign proliferative skin diseases, ichthyosis, papilloma, basal cell carcinoma, squamous cell carcinoma, restinosis, scleroderma and hemangioma.

The methods of the present invention may be used to treat any animal. Most preferably, the methods of the present invention are useful in humans. Generally, to achieve pharmacologically efficacious anti-proliferative effects, the p-heteroatom-substituted phenols and analogs may be administered in any therapeutically effective dose. Preferably, the p-heteroatom-substituted phenols and analogs are administered in a dose of from about 1 mg/kg to about 10 mg/kg.

A wide variety of p-heteroatom-substituted phenols and their analogs are effective in the methods of the present invention. Representative examples of p-heteroatom-substituted phenols and their analogs are compounds such those selected from the group comprising of _V-pivaloyl-5-hydroxy-3,4,6,7- tetramethylindole (27) , 4,6-di-tert-butyl-2,3-dihydro-5-hydroxy- 2(3i¥)-benzofuranone (24) 9-acetoxy-8,10-dimethyljulolidine (6), W-isobutyryl-5-acetoxy-3,4,6-trimethylindoline (12b) N- isobutyryl-5-methoxy-3,4,6-trimethylindoline (13), 5-hydroxy- 2,4,6,7-tetramethylbenzo-furan (33) and W-isobutyryl-5-hydroxy- 3,4,6-trimethylindoline (lib).

The term "individual" is meant to include animals and humans.

The term "biologically inhibiting" or "inhibition" of the growth of proliferating cells is meant to include partial or total growth inhibition and also is meant to include decreases in the rate of proliferation or growth of the cells. The

biologically inhibitory dose of the composition of the present invention may be determined by assessing the effects of the test element on target malignant or abnormally proliferating cell growth in tissue culture, tumor growth in animals and cell culture or any other method known to those of ordinary skill in the art.

Administration of the compositions of the present invention may be by topical, intraocular, parenteral, oral, intranasal, intravenous, intramuscular, subcutaneous, or any other suitable means. The dosage administered is dependent upon the age, clinical stage and extent of the disease or genetic predisposition of the individual, location, weight, kind of concurrent treatment, if any, and nature of the pathological or malignant condition. The effective delivery system useful in the method of the present invention may be employed in such forms as capsules, tablets, liquid solutions, suspensions, or elixirs, for oral administration, or sterile liquid forms such as solutions, suspensions or emulsions. Any inert carrier is preferably used, such as saline, or phosphate-buffered saline, or any such carrier in which the novel compounds used in the method of the present invention have suitable solubility properties.

Preferably, delivery systems useful in the method of the present invention may be employed in such sterile liquid forms such as solutions, suspensions or emulsions. For topical use it may be employed in such forms as ointments, creams or sprays. Any inert carrier is preferably used, such as saline, or phosphate-buffered saline, or any such carrier in which the compounds used in the method of the present invention have suitable solubility properties. There are a wide variety of pathological cancerous and noncancerous cell proliferative conditions for which the compositions and methods of the present invention will provide therapeutic benefits. These pathological conditions may occur in almost all cell types capable of abnormal cell proliferation. Among the cell types which exhibit pathological or abnormal growth are (1) fibroblasts, (2) vascular endothelial cells and (3) epithelial cells. It can be seen from the above that the

method of the present invention is useful in treating local or disseminated pathological conditions in all or almost all organ and tissue systems of the individual.

It is specifically contemplated that pharmaceutical compositions may be prepared using the novel p-heteroatom- substituted phenols, and analogs thereof, of the present invention. In such a case, the pharmaceutical composition comprises the novel p-heteroatom-substituted phenols, or analogs thereof, of the present invention and a pharmaceutically acceptable carrier. A person having ordinary skill in this art would readily be able to determine, without undue experimentation, the appropriate dosages and routes of administration of the novel p-heteroatom-substituted phenols and analogs of the present invention. In addition to the preferred antiproliferative activities of the novel p-heteroatom-substituted phenols and analogs of the present invention, these novel compounds also display novel cytoprotective, anti-mitogenic, angiostatic, anti- inflammatory, anti-atherosclerotic and anti-atherogenic activities. More preferably, the novel p-heteroatom-substituted phenols and analogs of the present invention are useful for various ocular reasons, e.g., aniostatic effects, inhibition of neovascularization, inhibition of cell proliferation following IOL implantation, inhibition of cataract formation (especially following vitrectomy) and heterodegenerative diseases, treatment for retinopathies, macular degeneration, photochemical injury, light damage injury (especially during ocular surgery) .

The novel p-heteroatom-substituted phenols and analogs of the present invention may be used as part of an eye irrigating solution. Preferably, such an irrigating solution would have the following formulation: active agent: 0.01-1.0%; cremephor EL: 0.5%; sodium chloride: 0.64%; potassium chloride: 0.075%; calcium chloride: 0.048%; magnesium chloride: 0.03%; sodium acetate: 0.39%; sodium citrate: 0.17%; sodium hydroxide and/or hydrochloric acid: as necessary to adjust pH; and water for administration.

The novel p-heteroatom-substituted phenols and analogs of the present invention may be used as part of an topical ocular preparation. Preferably, such an topical preparation would have the following formulation: active agent: 1.0%; polyvinyl alcohol: 1.4%; monobasic sodium phosphate monohydrate: 0.05%; dibasic sodium phosphate (anhydrous) 0.15%; sodium chloride: 0.5%; disodium EDTA: 0.01%; polysorbate 80: 0.01%; benzalkonium chloride solution: 0.01%; sodium hydroxide and/or hydrochloric acid: as necessary to adjust pH; and water for administration. Furthermore, the present invention shows that p- heteroatom-substituted phenolic compounds function as anti¬ oxidants under both biological and non-biological screening protocols. As an example, assays of anti-oxidant activities using a protocol involving retinal pieces showed IC _S0 's in the micromolar range.

The p-Heteroatom substituted phenolic compounds of the present invention possess antiproliferative properties, making them agents for prevention and treatment of proliferative diseases, including cancers of different cellular types and lineages.

Thus, the present invention is directed to an antiproliferative p-heteroatom-substituted phenol compound, or derivative thereof, having a structural formula selected from the group consisting of

wherein m is 0 to 3, n is 0 when Het is oxygen or sulfur, 0 to 4 when Het is nitrogen, Het is nitrogen, oxygen or sulfur; wherein R is selected from the group consisting of hydrogen, alkyl, arylmethyl and acyl; R ¹ is alkyl; R ² is selected from the

group consisting of hydrogen and alkyl; R ³ is selected from the group consisting of alkyl and acyl when Het is nitrogen, : (electron pair) when Het is oxygen or sulfur; R ⁴ is selected from the group consisting of hydrogen and alkyl; R ⁵ is selected from the group consisting of hydrogen and alkyl; and R ⁶ and R ⁷ are selected from the group consisting of hydrogen, alkyl and oxygen. The following examples are given for the purpose of illustrating various embodiments of the invention and are not meant to limit the present invention in any fashion. EXAMPLE 1

Synthesis of p-Heteroatom-substituted Phenols and Analogs Thereof

Melting points are uncorrected. Tetrahydrofuran was dried over benzophenone ketyl, toluene and methylene chloride were dried and distilled over calcium hydride. All air-sensitive and/or moisture-sensitive reactions were performed under a nitrogen or argon atmosphere. Concentration of solutions was by rotary evaporation unless otherwise noted.

IR spectra were obtained with a Nicolet 205 FT-IR spectrometer. Unless noted otherwise, ¹ H-NMR and ¹³ C-NMR spectra were recorded with a GE QE-300 MHz spectrometer using tetramethylsilane as internal standard and CDC1 ₃ as solvent. HRMS data were obtained on a VG ZAB-E (Fisons) high-resolution mass spectrometer using the El mode of ionization at 70 eV. Combustion analysis were performed by Atlantic Microlab Inc. EXAMPLE 2

Synthesis of 2.6-Dimethylanisole (2)

Sodium hydroxide (20 g, 500 mmol) was dissolved in distilled water (200 mL) and placed in a round-bottom flask fitted with a reflux condenser. To the solution were added sequentially 2,6-dimethylphenol (1, 61 g, 500 mmol) and dimethyl sulfate (48 mL, 500 mmol) . The mixture was heated at 60°C for about 17 h, cooled, and transferred to a separatory funnel. The organic layer was separated, and the aqueous layer was extracted with benzene. The combined organic layers were washed sequentially with 10% aq. sodium hydroxide and brine, and dried (MgSO . Concentration of the solution gave 2,6-dimethylanisole (2, 39 g, 57% yield), bp 179-182°C/750 torr (lit.: Burton, G.

W. ; Joyce, A.; Ingold, K. U. Arch. Biochem . Biophys . , 1982, 221 , 281; 181°C) , as a colorless oil.

EXAMPLE 3 Synthesis of 2.6-Dimethyl-4-nitroanisole ( 3 ) 2,6-Dimethylanisole (2, 36 g, 260 mmol) was added dropwise over 0.5 h to stirred cold cone, nitric acid (200 mL) contained in round-bottom flask; stirring at room temperature was continued for 18 h. The mixture was poured onto crushed ice contained in a 1-L beaker. The resulting bright yellow solid was vacuum-filtered and dried. Recrystallization from ethanol-water gave 2,6-dimethyl-4-nitroanisole (3, 29 g, 62% yield) as a pale yellow solid, mp 90-91°C (lit.: Burton, G. W. ; Joyce, A. ; Ingold, K. U. Arch . Biochem . Biophys . , 1982, 221 , 281; 89-91°C) .

EXAMPLE 4 Synthesis of 4-Amino-2.6-dimethylanisole ( 4 )

2,6-Dimethyl-4-nitroanisole (3, 29 g, 160 mmol), 10% Pd-C (200 mg) and ethanol (300 mL) were placed in a 500-mL Parr hydrogenation flask and hydrogenated at 50 psi for 24 h. The resulting solution was filtered through a silica gel column using ethyl acetate as eluent. Concentration of the solvent and drying under vacuum gave 23.7 g (98% yield) of 4-amino-2,6- dimethylanisole (4) as a brown-colored solid, mp 59-60°C (lit.: Bruice, T. C; Kharasch, N. ; Winzler, R. J. J. Org. Chem . 1953, 18, 83; 63°C) . EXAMPLE 5

Synthesis of 9-Methoxy-8.10-dimethyliulolidine f5)

4-Amino-2,6-dimethylanisole (4, 1.4 g, 9.3 mmol), freshly distilled 1,3-bromochloropropane (13.7 mL, 139 mmol), anhydrous sodium carbonate (3.9 g, 37 mmol) and molecular sieves (4 A, 1 g) were placed in a round-bottom flask and, under an argon atmosphere, stirred at 70°C for 1 h, at 100°C for 2 h and then heated under reflux for 17 h. The cooled reaction mixture was filtered and the filtrate was acidified with cone, hydrochloric acid. 1,3-Bromochloropropane was removed by steam distillation, and the residue was basified with aq. sodium hydroxide and extracted with ether. The combined extracts were washed with water and brine. Removal of solvents after drying

(MgS0 ₄ ) gave a crude material which was purified by flash column chromatography, using 3:7 ethyl acetate:hexanes as eluent, to yield 9-methoxy-8,10-dimethyljulolidine (5, 1.2 g, 56% yield) as a yellow solid, mp 55-57.5°C. Spectral data: ^H NMR: δ 2.0 (m, 4H) , 2.12 (S, 6H) , 2.63 (t, 4H, J = 6.9 Hz), 3.0 (t, 4H, J = 4.5 Hz) 3.61 (s, 3H) ; HRMS: calcd. for C ₁₅ H ₂₁ NO 231.1623, fnd. 231.1618.

EXAMPLE 6 Synthesis of 9-Hvdroxy-8.10-dimethyliulolidine hvdrobromide 9-Methoxy-8,10-dimethyljulolidine (5, 109 mg, 0.5 mmol) and 49% hydrobromic acid (2 L) were placed in a round-bottom flask and refluxed for 3 h under an argon atmosphere. The reaction mixture was cooled to room temperature and washed several times with water and then twice with acetone. Drying under vacuum gave julolidine hydrobromide (70 mg, 47% yield) as a colorless solid, mp 235-237°C. Spectral data: ¹ H NMR (CD ₃ OD) : δ 2.05 (s, 6H) , 2.18 (q, 4H, J = 6.8 Hz), 2.76 (t, 4H, J = 6.6 HZ), 4.8 (s, 4H) ; ¹³ C NMR (CD ₃ 0D) : δ 11.7, 18.7, 23.4, 51.7, 121.8, 125.0, 127.2, 152.0. Anal: calcd. for C _u H ₂₀ BrNO C, 56.39, H, 6.76, N, 4.70, fnd. C, 56.09, H, 6.69, N, 4.67.

EXAMPLE 7 Synthesis of 9-Acetoxy-8.10-dimethyliulolidine (6)

9-Hydroxy-8,10-dimethyljulolidine hydrobromide (832 mg, 2.79 mmol) and methylene chloride (4 L) were placed in a round- bottom flask and stirred under an argon atmosphere.

Triethylamine (1.67 mL, 12 mmol) and acetic anhydride (0.55 mL, 5.8 mmol) were added sequentially, and the mixture was stirred at room temperature for about 16 h. The mixture was poured into water (15 mL) and extracted with methylene chloride. The combined methylene chloride extracts were washed sequentially with 5% hydrochloric acid and brine and dried (Na ₂ SO,,) . Removal of solvents furnished a solid which was purified by flash column chromatography using 1:5 ethyl acetate:hexanes as eluent to yield 9-acetoxy-8,10-dimethyljulolidine (6, 680 mg, 94% yield), mp 112-113°C. Spectral data: ¹ H NMR: δ 1.90 (s, 6H) , 1.95 ( , 4H) , 2.30 (s, 3H) , 2.64 (t, 4H, J = 7 Hz), 3.02 (t, 4H, J = 5 Hz); ¹³ C NMR: S 12.3, 20.5, 22.3, 25.2, 50.0, 119.0, 125.3,

139.0, 142.0, 169.9. Anal: calcd. for C ₁₆ H ₂ ,N0 ₂ C, 74.10, H, 8.16, N, 5.41, fnd. C, 73.90, H, 8.20, N, 5.36.

EXAMPLE 8 Synthesis of 4-Acetamido-2.6-dimethylanisole C7) 4-Amino-2,6-dimethylanisole (4, 5 g, 33.2 mmol) was placed in a round-bottom flask fitted with a reflux condenser, and acetic anhydride (3.4 mL, 35.4 mmol) and acetic acid (3 mL, 53.1 mmol) were added sequentially. The mixture was heated to 50°C for 4 h, cooled, poured into water and basified with saturated aq. sodium bicarbonate. The solution was extracted with methylene chloride, and the combined extracts were washed with brine and dried (MgS0 ₄ ) . Removing the solvent and recrystallizing the resulting solid from hexane-methylene chloride yielded 4-acetamido-2,6-dimethylanisole (7, 5.3 g, 83% yield). Spectral data: ¹ H NMR: δ 2.1 (s, 3H) , 2.2 (s, 6H) , 3.65 (s, 3H) , 7.10 (s, 2H) , 8.10 (s, IH) ; ¹³ C NMR: δ 16.1, 24.2, 60.0, 121.5, 131.8, 134.3, 154.3, 169.8; CI-HRMS: calcd. for C^H^NO _g (M+H ⁺ ) 194.1181, fnd. 194.1181.

EXAMPLE 9 Synthesis of 4-Acetamido-3-bromo-2,6-dimethylanisole

4-Acetamido-2,6-dimethylanisole (7, 8.6 g, 44.3 mmol) and acetic acid (30 mL) were placed in a round-bottom flask under an argon atmosphere. Bromine (2.7 mL, 44.3 mmol) was added dropwise to the solution, after which the reaction mixture was stirred at room temperature for about 3 h, and then concentrated. The residue was diluted with methylene chloride and carefully washed sequentially with saturated aq. sodium bicarbonate and brine, and dried (MgSO . Removal of solvents gave a brown- colored solid which was recrystallized from methylene chloride- hexanes to yield 4-acetamido-3-bromo-2,6-dimethylanisole (10.3 g, 85% yield) as an off-white solid, mp 152-153°C. Spectral data: ¹ H NMR: δ 2.18 (s, 3H) , 2.24 (s, 3H) , 2.35 (ε, 3H) , 3.66 (S, 3H) , 7.55 (br s, IH) , 7.9 (s, IH) ; ¹³ C NMR: δ 16.1, 16.8, 24.6, 60.1, 114.5, 122.0, 130.5, 131.3, 131.5, 153.6, 168.1; CI- HRMS: calcd. for C ₁₁ H ₁₅ BrN0 ₂ (M+H ⁺ ) 272.0286, fnd. 272.0284.

EXAMPLE 10

Synthesis of jV-Allyl-4-acetamido-3-bromo-2.6-dimethylanisole (8. Sodium hydride (1.1 g, 27 mmol, 60% dispersion in oil) contained in a round-bottom flask fitted with a reflux condenser was washed using dry hexanes, tetrahydrofuran (50 mL) was added, and the suspension was stirred at 0°C under a nitrogen atmosphere. A solution of 4-acetamido-3-bromo-2,6- dimethylanisole (7.3 g, 27 mmol) dissolved in tetrahydrofuran (50 mL) was added dropwise using a canula over about 0.5 h. Allyl bromide (4.85 mL, 84.8 mmol) was added to the cold solution in one portion, and the mixture was warmed to 60°C and held there for 17 h. The reaction mixture was quenched with saturated aq. ammonium chloride and extracted with diethyl ether. The combined extracts were washed with brine and dried (MgSO . Removal of solvents and purification of the resulting viscous liquid by flash column chromatography using 1:3 ethyl acetate:hexanes as eluent gave N-allyl-4-acetamido-3-bromo-2,6-dimethylanisole (8, 8.7 g, 85% yield) as a viscous liquid. Spectral data: ¹³ C NMR: δ 15.2, 16.3, 21.5, 50.1, 59.3, 117.1, 123.3, 129.2, 130.2, 132.37, 132.4, 136.5, 156.2, 169.1; HRMS: calcd. for C _u H ₁₉ BrN0 ₂ (M+H ^* ) 312.0599, fnd. 312.0586.

EXAMPLE 11 Synthesis of W-Acetyl-5-methoχy-3,4.6-trimethylindoline (9 )

A solution of W-allyl-4-acetamido-3-bromo-2,6- dimethylanisole (8, 1.1 g, 3.6 mmol) in dry toluene (237 mL) and tri-n-butylstannane (1.9 mL, 7.1 mmol) was placed in a round- bottom flask fitted with a reflux condenser and slowly warmed to 60°C under a nitrogen atmosphere. A catalytic amount of azobisisobutyronitrile was added, and the resulting mixture was heated and stirred at 85°C for 15 h. The cooled reaction mixture was washed sequentially with 1% aq. ammonia and brine, and dried (MgS0 ₄ ) . Removal of solvents gave a liquid which was further purified by flash column chromatography using sequentially hexane and 1:3 ethyl acetate:hexanes as eluents to yield the induline 9 (790 mg, 95% yield) as a colorless liquid. Spectral data: ¹³ C NMR: δ 11.9, 16.4, 20.5, 23.9, 33.9, 57.3, 59.8, 116.5, 126.3,

129.7, 133.9, 137.6, 153.1, 168.2; HRMS: calcd. for C _H H _2fJ N0 ₂ (M+H*) 234.1494, fnd. 234.1487.

EXAMPLE 12 Synthesis of iV-Acetyl-5-hydroxy-3.4.6-trimethylindoline (Ila) Boron tribromide (1.4 mL of 1 M solution in methylene chloride, 1.4 mmol) was added to a cooled (-78°C) solution of induline 9 (110 mg, 47 mmol) dissolved in methylene chloride (2 L) . The resulting solution was maintained at -78°C (1 h) , then slowly warmed to room temperature and held there for 17 h. The cooled reaction mixture was quenched with a few drops of methanol followed by cold water. The solution was extracted with methylene chloride and the combined organic layers were washed with brine and dried (MgS0 ₄ ) . Removal of solvents gave a solid, which was recrystallized from methylene chloride-hexanes to afford the induline Ila (95 mg, 92% yield) as a colorless solid, mp 204-205°C. Spectral data: ¹³ C NMR (CD ₃ SOCD ₃ ) : δ 12.3, 17.2, 20.7, 23.7, 33.4, 56.4, 115.3, 120.7, 122.6, 133.9, 134.3, 149.1, 167.4; HRMS: calcd. for C ₁₃ H ₁₇ N0 ₂ 219.1259, fnd. 219.1261. Anal: calcd. for C ₁₃ H ₁₇ N0 ₂ C, 71.21, H, 7.81, N, 6.39, fnd. C, 71.06,H, 7.84, N, 6.33.

EXAMPLE 13 Synthesis of JV-Acetyl-5-acetoxy-3. .6-trimethylindoline C12a)

To an ice-cold solution of induline Ila (219 mg, 1 mmol) dissolved in methylene chloride (5 mL) were added sequentially pyridine (1 mL) and acetic anhydride (0.12 mL, 1.1 mmol) , and the resulting mixture was stirred over a range of 0°C to room temperature for 18 h. The reaction mixture was quenched with water and extracted with methylene chloride. The combined organic extracts were washed with 5% aq. hydrochloric acid and brine then dried (MgSO . Removal of solvents and chromatographic purification of the residue using 3:7 ethyl acetate:hexanes as eluent gave the acetate 12a (228 mg, 87% yield) as a pale yellow solid. This was recrystallized from hexanes-methylene chloride to afford a colorless solid, mp 128-130°C. Spectral data: ¹ H NMR: δ 1.23 (d, 3H, J = 7 Hz) , 2.04 (s, 3H) , 2.10 (s, 3H) , 2.17 (s, 3H) , 2.31 (s, 3H) , 3.61 (dd, IH, J = 10.2, 2.3 HZ), 4.13 (t, IH, J = 9.53 Hz) , 7.96 (s, IH) ;

¹³ C NMR: δ 12.3, 16.7, 20.4, 20.7, 24.8, 33.9, 57.4, 116.4, 125.7, 129.3, 133.7, 139.5, 144.2, 168.5, 169.1; HRMS: calcd. for C ₁₅ H _2fJ N0 ₃ (M+H ⁺ ) 262.1443, fnd. 262.1446.

EXAMPLE 14 Synthesis of 5-Methoxy-3.4.6-trimethylindoline

To an ice-cold solution of induline 9 (377 mg, 1.6 mmol) dissolved in tetrahydrofuran (5 mL) and kept under an argon atmosphere was added lithium triethylborohydride (3.2 mL, 3.2 mmol) , and the mixture was stirred over the range of 0°C to room temperature for 20 h. The reaction mixture was quenched with 3 N hydrochloric acid, and volatiles were removed under vacuum. The resulting solution was acidified to pH 2 with additional 3 N hydrochloric acid and washed with methylene chloride. The aqueous solution was basified with saturated aq. sodium bicarbonate and extracted with methylene chloride. The organic extracts were washed with brine and dried (MgSO . Removal of solvents gave crude methoxyindoline (200 mg, 65% yield) as a brownish yellow oil, which was used without further purification.

EXAMPLE 15 Synthesis of 5-Hvdroxy-3. 4 ,6-trimethylindoline (10)

The methoxyindoline (185 mg, 0.96 mmol) and 49% hydrobromic acid (4 mL) were placed in a round-bottom flask fitted with a reflux condenser and heated to 100°C for 3 h under a nitrogen atmosphere. The solution was cooled, diluted with water, and washed off with ether. The acidic solution was basified with saturated aq. sodium bicarbonate and extracted with methylene chloride. The combined organic extracts were washed with brine and dried (MgS0 ₄ ) . Removal of solvents gave a brown- colored oil which was purified by filtration through a silica gel plug using 1:1 ethyl acetate:hexanes as eluent to afford induline 10 (160 mg, 94% yield) as a yellow oil. This material was immediately used in the next step.

EXAMPLE 16 Synthesis of iV-lsobutyryl-5-hvdroxy-3.4,6-trimethylindoline (lib) To an ice-cold solution of induline 10 (160 mg, 0.9 mmol) dissolved in methylene chloride (4 mL) and kept under an argon atmosphere were added sequentially pyridine (1 mL) and

isobutyryl chloride (0.1 mL, 9 mmol); the mixture was stirred over the range of 0°C to room temperature for some 17 h. The mixture was diluted with methylene chloride, washed with brine and dried (MgS0 ₄ ) . Removal of solvents followed by purification of the residue using 1:9 ethyl acetate:hexanes as eluent afforded the desirediV-isobutyryl-5-hydroxy-3,4,6-trimethylindoline (lib, 163 mg, 73% yield) as a colorless solid, mp 123-125°C. Spectral data: 'H NMR: δ 1.1-1.25 (m, 9H) , 2.17 (s, 3H) , 2.20 (s, 3H) , 2.72 (quintet, IH, J = 6.5 Hz), 3.35 (t, IH, J = 6.5 Hz) , 3.78 (dd, IH, J = 10.1, 1.0 Hz), 4.13 (t, IH, J = 9.1 Hz) , 7.95 (s, IH) ; ¹³ C NMR: δ 11.9, 16.5, 18.8, 19.3, 20.7, 33.1, 34.0, 56.3, 116.8, 119.5, 122.1, 134.1, 135.0, 148.8, 175.0; HRMS: calcd. for C ₁₅ H ₂₁ N0 ₂ 247.1523, fnd. 247.1573. Anal: calcd. for C ₁₅ H ₂₁ N0 ₂ C, 72.85, H, 8.55, N, 5.66, fnd. C, 72.63, H, 8.59, N, 5.60. EXAMPLE 17

Synthesis of iV-Isobutyryl-5-acetoxy-3 _f 4.6-trimethylindoline C12b)

To an ice-cold solution of W-isobutyryl-5-hydroxy-

3,4,6-trimethylindoline (lib, 49 mg, 0.2 mmol) dissolved in methylene chloride (2 mL) were added sequentially pyridine (0.16 mL, 2 mmol) and acetic anhydride (0.1 mL, 1 mmol). The mixture was stirred over the range of 0°C to room temperature for 17 h. The mixture was diluted with methylene chloride, transferred to a separatory funnel and washed sequentially with 10% hydrochloric acid, saturated aq. sodium bicarbonate and brine. Removal of solvents after drying (MgS0 ₄ ) gave a viscous liquid, which was purified by flash column chromatography, using 1:4 ethyl acetate:hexanes as eluent, to afford the W-isobutyryl-5-acetoxy- 3,4,6-trimethylindoline (12b, 45 mg, 77% yield) as a colorless solid, mp 118-120°C. Spectral data: ¹ H NMR: δ 2.0 (s, 3H) , 2.06 (s, 3H) , 2.26 (s, 3H) , 2.7 (quintet, IH, J = 6.7 Hz), 3.3 (t, IH, J = 6.7 Hz), 3.7 (dd, IH, J = 10.2, 1.2 Hz), 4.1 (t, IH, J = 9.3 Hz), 8.0 (s, IH) ; CI-HRMS: calcd. for C ₁₇ H ₂₄ N0 ₃ (M+H ⁺ ) 290.1756, fnd. 290.1756.

EXAMPLE 18 Synthesis ofiV-Isobutyryl-5-methoxy-3.4.6-trimethylindoline (13) Sodium hydride (7 mg, 0.18 mmol ) contained in a round- bottom flask was washed using dry hexanes and tetrahydrofuran (2

mL) was added, and the suspension was stirred at 0°C under nitrogen atmosphere. A solution of _V-isobutyryl-5-hydroxy-3,4,6- trimethylindoline (lib, 42 mg 0.17 mmol) dissolved in tetrahydrofuran (2 mL) was added dropwise using a syringe. Methyl iodide (0.2 mL) was added to the cold solution in one portion, and the mixture was heated under reflux for 17 h. The reaction mixture was cooled, quenched with saturated aq. ammonium chloride solution and extracted with ether. The combined extracts were washed with brine and dried (MgS0 ₄ ) . Removal of solvents gave a viscous liquid which was purified by flash column chromatography using 1:4 ethyl acetate:hexanes as eluent, to affordN-isobutyryl-5-methoxy-3,4,6-trimethylindoline (13, 40mg, 90% yield) as a viscous liquid. Spectral data: ¹ H NMR: δ 1.20-1.35 ( , 9H) , 2.22 (s, 3H) , 2.27 (s, 3H) , 2.74 (quintet, IH, J = 6.5 Hz), 3.30 (t, IH, J - ^* 6.5 Hz) , 3.65 (s, 3H) , 3.76 (dd, IH, J = 10, 1 Hz), 4.15 (t, IH, J = 9 Hz) , 8.0 (s, IH) .

EXAMPLE 19 Synthesis of iV-Pivaloyl-5-hvdroxy-3.4.6-trimethylindoline flic) Treatment of induline 10 (160 mg, 0.9 mmol) according to a procedure similar to one used for preparing lib, with substitution of pivaloyl chloride for isobutyryl chloride, afforded lie (176 mg, 75% yield) as a colorless solid, mp 144-145°C. Spectral data: ¹³ C NMR: δ 11.7, 16.4, 19.5, 27.6, 39.9, 57.4, 117.9, 119.2, 121.9, 133.7, 136.0, 148.8, 176.0; HRMS: calcd. for C ₁₆ H ₂₃ N0 ₂ 261.1229, fnd. 261.1921. Anal: calcd. for C ₁₆ H ₂₃ N0 ₂ C, 73.53, H, 8.87, N, 5.36, fnd. C, 73.60, H, 8.91, N, 5.36.

EXAMPLE 20 Synthesis of JV-C2-Methyl-2-propenyl)-4-acetamido-3-bromo-2.6- dimethylanisole (14)

Treatment of 4-acetamido-3-bromo-2,6-dimethylanisole (2.7 g, 10 mmol) according to a procedure similar to the one used for making compound 8, substituting methallylbromide for allyl bromide, afforded 14 (2.8 g, 86% yield) as a viscous liquid. Spectral data: ¹ H NMR (250 MHz): δ 1.71 (s, 3H) , 1.75 (s, 3H) , 2.18 (s, 3H) , 2.33 (s, 3H) , 3.26 (d, IH, J = 15 Hz), 3.66 (s,

3H) , 4.62 (s, IH) , 4.75 (s, IH) , 4.82 (d, IH, J = 15 Hz) , 6.82 (S, IH).

EXAMPLE 21 General Procedure for Palladium-catalyzed Cvclization of Bromoanilides to Produce Indoles and Indolines

Palladium (II) acetate (0.1 eq.), triphenylphosphine (0.2 eq.), tetraethylammonium chloride (1 eq.), sodium formate (1.1 eq.) and dimethyl formamide were placed in a round-bottom flask fitted with a reflux condenser, and the mixture was stirred under a nitrogen atmosphere. A solution of anilide dissolved in dimethyl formamide was added, and the mixture was heated at either 85 or 100°C for several hours, until TLC analysis showed consumption of all precursor. The mixture was cooled, gravity- filtered and the filter cake was washed with diethyl ether. Concentration followed by distillation of the dimethyl formamide under vacuum gave a viscous liquid, which was purified by flash column chromatography

EXAMPLE 22 Synthesis ofW-Acetyl-5-methoxy-3.3.4,6-tetramethylindoline (15) Treatment of 14 (1.9 g, 5.8 mmol) according to the general procedure used for Pd-catalyzed cyclization afforded 15 (1.2 g, 85% yield) as a viscous liquid. ¹³ C NMR: «5" 11.6, 16.4, 24.1, 26.9, 41.1, 59.9, 64.7, 116.7, 126.5, 129.7, 135.7, 137.8, 153.4, 168.0; HRMS: calcd. for C ₁₅ H ₂₁ N0 ₂ 247.1572, fnd. 247.1581. EXAMPLE 23

Synthesis of iV-Acetyl-5-hydroxy-3.3.4.6-tetramethylindoline (16)

Treatment of 15 (741 mg, 3 mmol) according to a procedure similar that used for making Ila afforded 16 (524 mg,

75% yield) as a colorless solid, mp 173-174°C. Spectral data: ¹³ C NMR (CD ₃ SOCD ₃ ) : δ 11.7, 17.3, 23.9, 27.0, 40.6, 63.7, 115.4, 121.1, 122.8, 135.6, 149.2, 167.2; HRMS: calcd. for C _u H, ₉ N0 ₂ 233.1415, fnd. 233.1416. Anal: calcd. for C ₁₄ H ₁₉ N0 ₂ C, 72.08, N, 6.00, H, 8.20, fnd. C, 71.88, N, 5.95, H, 8.22.

EXAMPLE 24 Synthesis of W-Pivaloyl-4-amino-2.6-dimethylanisole

Into an ice-cold solution of aniline 4 (15.1 g, 100 mmol) dissolved in methylene chloride (100 mL) were added

sequentially pyridine (17.5 mL) and pivaloyl chloride (12.3 mL, 100 mmol) dropwise, and the resulting mixture was stirred at room temperature for 15 h. The mixture was diluted with methylene chloride and washed sequentially with 10% aq. hydrochloric acid, saturated aq. sodium bicarbonate and brine. Removal of solvents after drying (MgS0 ₄ ) gave the pivalamide (19.8 g, 86% yield) as a pale brown solid, mp 126-128°C. Spectral data: ¹ H NMR: <S 1.30 (S, 9H) , 2.23 (s, 6H) , 3.68 (s, 3H) , 7.18 (s, 2H) .

EXAMPLE 25 Synthesis of W-Pivaloyl-4-amino-3-bromo-2.6-dimethylanisole (17)

Into an ice-cold solution of the pivalanilide (19.8 g,

85.7 mmol) dissolved in acetic acid (100 mL) was added bromine

(4.4 mL, 85.7 mmol) dropwise using a syringe, and the resulting solution was stirred at room temperature for 19 h. The mixture was diluted with methylene chloride, transferred to a beaker and carefully basified with saturated aq. sodium bicarbonate. The biphasic solution was transferred to a separatory funnel and extracted with methylene chloride. The combined methylene chloride layers were dried (MgSO _A ) and concentrated to give a brown-colored solid which was recrystallized from methylene chloride-hexane to afford 119 (23.5 g, 87% yield) as a pale brown solid, mp 87-88°C. Spectral data: ^ NMR: tS 1.32 (s, 9H) , 2.22 (s, 3H) , 2.36 (s, 3H) , 3.65 (s, 3H) , 7.90 (br s, IH) , 8.05 (s, IH) . EXAMPLE 26

Synthesis of N-(2-Methv1-2-propenyl) . λ ^τ -Pivaloyl-4-amino-3-bromo- 2.6-dimethylanisole (18)

Sodium hydride (1.5 g, 38.3 mmol) contained in a round- bottom flask fitted with a reflux condenser was washed using dry hexanes, tetrahydrofuran (50 mL) was added and the suspension was stirred at 0°C under a nitrogen atmosphere. A solution of compound 119 (12 g, 38.3 mmol) dissolved in tetrahydrofuran (70 mL) was added by a cannula over 0.5 h. Methallyl bromide (6 mL, 59.5 mmol) was added dropwise using a syringe, and the mixture was slowly heated to 60°C. Stirring and heating were continued for 18 h. The mixture was quenched with saturated aq. ammonium chloride and extracted with ether. The combined ethereal layers

were washed with brine and dried (MgS0 ₄ ) . Removal of the solvents and purification of the residue by filtering through a small silica gel column using 1:3 ethyl acetate-hexane as the eluent gave 18 (13 g, 93% yield) as a pale yellow liquid. Spectral data: ¹ H NMR: δ 1.00 (s, 9H) , 1.70 (s, 3H) , 2.21 (s, 3H) , 2.34 (S, 3H) , 3.39 (d, IH, J = 13 Hz), 3.68 (s, 3H) , 4.65 (br S, IH) , 4.77 (br s, IH) , 4.99 (d, IH, J = 15.2 Hz) , 6.93 (s, IH) ; ¹³ C NMR: δ 16.0, 17.1, 20.7, 28.8, 41.0, 56.8, 60.2, 112.0,

125.6, 129.8, 130.7, 133.0, 138.3, 140.8, 156.8, 177.3; HRMS: calcd. for C ₁₈ H ₂₇ BrN0 ₂ (M+H ^* ) 368.1225, fnd. 368.1213.

EXAMPLE 27 Synthesis of iV-Pivaloyl-3.3,4,6-tetramethyl-5-methoxyindoline

(19)

Treatment of 18 (5.1 g, 14 mmol) according to the general procedure used for Pd-catalyzed cyelization afforded 19 (2.1 g, 52% yield) as a colorless solid, mp 103-104°C. Spectral data: H NMR: δ 1.32 (s, 9H) , 1.38 (s, 6H) , 2.30 (s, 3H) , 2.33 (S, 3H) , 3.65 (s, 3H) , 3.85 (s, 2H) , 7.95 (s, IH) ; ¹³ C NMR: δ 11.5, 16.5, 25.8, 27.6, 39.9, 41.8, 59.9, 64.6, 118.3, 126.2, 129.4, 135.7, 139.4, 153.4, 175.9.

EXAMPLE 28 Synthesis of 5-Hvdroxy-3.3.4,6-tetramethylindoline hydrobromide In a round-bottom flask fitted with a reflux condenser were placed 19 (6 g, 20.8 mmol) and 49% hydrobromic acid (100 mL) , and the mixture was heated at 100°C for 18 h under a nitrogen atmosphere, and then cooled. The hydrobromic acid was removed under reduced pressure, and the resulting solid was washed with hexanes to yield 5-hydroxy-3,3,4,6- tetramethylindoline hydrobromide (5.2 g, 92% yield) as a colorless solid. Spectral data: ¹ H NMR (CD ₃ OD) : <S 1.50 (s, 6H) , 2.25 (s, 3H) , 2.30 (s, 3H) , 3.50 (s, 2H) , 7.00 (s, IH) ; ¹³ C NMR (CD ₃ OD) : <S 12.0, 17.1, 26.6, 44.6, 59.9, 119.2, 123.2,

126.7, 127.1, 140.6, 156.2.

EXAMPLE 29 Synthesis of iV-Pivaloyl-5-hydroxy-3.3.4.6-tetramethylindoline 1201

Treatment of 5-hydroxy-3,3,4,6-tetramethylindoline hydrobromide (218 mg, 0.8 mmol) according to a procedure similar to that used for making >V-isobutyryl-5-hydroxy-3,4,6- trimethylindoline (lib) afforded 20 (155 mg, 70% yield) as a colorless solid, mp 143-144°C. Spectral data: ¹³ C NMR: δ 11.3, 16.4, 26.1, 27.7, 39.9, 41.9, 64.6, 118.1, 119.3, 121.3, 135.9, 136.7, 148.9, 175.6; CI-HRMS: calcd. for C ₁₇ H ₂₆ N0 ₂ (M+H ⁺ ) 276.1964, fnd. 276.1955. Anal: calcd. for C ₁₇ H ₂₅ N0 ₂ C, 74.15, H, 9.15, N, 5.09, fnd. C, 73.94, H, 9.21, N, 5.04.

EXAMPLE 30 Synthesis of 2 ,6-Di-tert-butyl-l-4-hvdroquinone 2,6-Di-tert-butyl-l,4-benzoquinone (5.0 g, 22.6 mmol), prepared according to the method of Minisu, et al., J. Org. Chem.. 54:728,(1989) was dissolved in diethyl ether (40 mL) and shaken vigorously in a separatory funnel with a solution of sodium dithionite (8 g, 33 mmol) in water (60 mL) . The color of the organic layer changed from bright mustard to brown and finally to pale yellow. This layer was washed with brine and dried (Na ₂ S0 ₄ ) . Concentration afforded 4.6 g (92% yield) of a peach-colored solid, mp 103-106°C. Spectral data: ¹ H NMR: δ 1.45 (s, 18H) , 4.80 (br s, IH) , 5.45 (br s, IH) , 6.20 (s, 2H) ; ¹³ C NMR: δ 30.13, 34.34, 111.97, 137.49, 147.62, 147.90.

EXAMPLE 31 Synthesis of 5-AlIyloxy-l.3-di-tert-butylphenol

A solution of 2,6-di-tert-butyl-l,4-hydroquinone (5.8 g, 26 mmol) in tetrahydrofuran (50 mL) was added dropwise to a suspension of sodium hydride in tetrahydrofuran at o°C This mixture was stirred for 15 minutes and then allyl bromide (3.47 g, 27.7 mmol) was added dropwise over 3-5 min. The solution was allowed to warm to room temperature and left stirring for 16 h. It was then quenched with ice water, extracted into diethyl ether, washed with brine and dried (Na ₂ S0 ₄ ) . Concentration afforded a crude dark oil which was purified by flash column chromatography over silica gel using a solvent system of hexanes

and ethyl acetate (95/5, R _f = 0.42) and resulted in 4.40 g of a yellow liquid (64% yield). Spectral data: ¹ H NMR: δ 1.42 (s, 18H) , 4.44 (d, 2H, J ^• = 5.7 Hz), 4.78 (br s, IH) , 5.28 (d, IH, J = 10.3 Hz), 5.42 (d, IH, J = 16.2 Hz) , 6.02-6.16 (m, IH) , 6.78 (s, 2H) ; ¹³ C NMR: -530.20, 34.57, 69.54, 111.56, 117.38, 134.07, 137.15, 147.84, 151.57.

EXAMPLE 32 Synthesis of 5-Al 1y1oxy- 1.3 -di -tert-butv1-2 - trimethylsiloxybenzene (21) 5-Allyloxy-l,3-di-tert-butylphenol (0.03 mmol) andbis- trimethylsilylacetamide (BSA, 15.5 g, 0.076 mol) were dissolved in dry acetonitrile (30 mL) . The resulting solution was refluxed for 24 h and then poured into a beaker containing ice water (50 mL) . The solution was extracted with diethyl ether (3 x 20 mL) , washed with brine and dried (Na ₂ S0 ₄ ) . Concentration afforded 10.2 g of a tawny brown liquid. This crude material was rapidly passed through a 5 cm-long pad of silica gel (90-mm diameter) with 5% ethyl acetate in hexanes to yield 8.4 g of liquid product. Spectral data: ¹ H NMR: δ 0.40 (s, 9H) , 1.40 (s, 18H) , 4.48 (d, 2H) , 5.28 (dd, IH) , 5.42 (dd, IH) , 6.02-6.16 (m, IH) , 6.84 (s, 2H) .

EXAMPLE 33 Synthesis of 2-Allyl-3.5-di-tert-butyl-4-trimethylsiloxyphenol (22a) Phenol 22 was prepared by BCl ₃ -assisted rearrangement of 5-allyloxy-l,3-di-tert-butyl-2-trimethylsiloxybenzene (21) according to Gilbert and Pinto, J. Org. Chem . 1992, 57 , 5271, for thepreparationof 2-allyl-3,5,6-trimethyl-l,4-hydroquinone, with the following modifications: the ratio of substrate to BC1 ₃ was 1:1.1 and the reaction was quenched at -70°C by the addition of a saturated aqueous solution of sodium carbonate and by allowing the resulting mixture to reach ambient temperatures. The crude product was isolated as a viscous green oil (81% yield) and acetylated without further purification. Spectral data: H NMR: δ 0.18 (s, 9H) , 1.34 (s, 9H) , 1.53 (s, 9H) , 3.63 (d, 2H, J = 5.2 HZ), 5.10 (dd, IH, J = 17.3, 2 Hz) , 5.20 (dd, IH, J = 10.2, 2 HZ), 5.92-6.08 (m, IH) , 6.68 (s, IH) .

EXAMPLE 34 Synthesis of 1-Acetoxy-2-allyl-3.5-di -tert-butyl-4- trimethylsiloxybenzene (22b)

Compound 22b was prepared by the acetylation of 2- allyl-3, 5-di-tert-butyl-4-trimethylsiloxyphenol (22a) according to the method of Gilbert and Pinto, J . Org . Chem . 1992, 57 , 5271- 5276. Spectral data: ¹ H NMR: δ 0.18 (ε, 9H) , 1.36 (s, 9H) , 1.23 (S, 9H) , 2.21 (s, 2H) , 3.55 (d, 2H) , 4.90 (dd, IH) , 4.98 (dd, IH) , 5.77-5.90 (m, IH) , 6.78 (s, IH) ; EI-MS: m/z 376 (M ^* ) , 334 (base, M* - CH ₂ CO) , 319, 73, 57; HRMS: Calcd. for C ₂₂ H ₃₆ 0 ₃ Si 376.2434, fnd. 376.2430.

EXAMPLE 35

Synthes is of 6'-Acetoxy-2' .4'-di-tert-butγl-3'- trimethylsiloxyphenylacetaldehvde (23a) Ozonolysis of 22b according to the procedure of Gilbert and Pinto, J. Org. Chem . 1992, 57 , 5271, for the ozonolysis of 2-allyl-l,4-diacetoxy-3 , 5,6-trimethylbenzene afforded 23a, which was obtained as a white solid (50% yield) after flash column chromatography over silica gel (Skellysolve B-ethyl acetate, 90:10, R = 0.23). Spectral data: ¹ H NMR: δ 0.22 (s, 9H) , 1.37 (s, 9H) , 1.44 (s, 9H) , 2.20 (s, 2H) , 3.74 (d, 2H, J =1.6 Hz), 6.88 (s, IH) , 9.52 (t, IH, J = 1.7 Hz) ; HRMS: calcd. for C ₂₁ H ₃₄ SiO 378.222G, fnd. 378.2214.

EXAMPLE 36 Synthesis of 6'-Acetoxy-2' ,4'-di-tert-butyl-3'- trimethylsiloxyphenylacetic acid (23b)

Aldehyde 23a was oxidized to 23b according to the procedure of Gilbert and Pinto, J . Org. Chem . 1992, 57 , 5271. The acid was obtained as a white solid (88% yield) . Spectral data: 'H NMR: δ 0.20 (ε, 9H) , 1.35 (s, 9H) , 1.45 (s, 9H) . 2.23 (s, 3H) , 3.80 (s, 2H) , 6.90 (s, IH) , carboxylic acid proton was not detected. El MS: m/z 394 (M ^* , very low intensity) , 352 (M ^* - CH ₂ CO) , 334 (base, 352 - H ₂ 0) , 319, 291, 263, 73, 57, 45.

EXAMPLE 37 Synthesis of 4.6-Di-tert-butyl-2.3-dihydro-5-hvdroxy-2 (3H)-benzo- furanone (24)

With rigorous exclusion of oxygen from the reaction vessel, a solution of 23b (2.30 g, 5.8 mmol) in methanol (15 mL) was sparged with argon for 20 min. Potassium carbonate (0.80 g, 5.8 mmol) was added and the resulting mixture was allowed to stir for 3 hours, after which time the pH of the reaction mixture was brought to 3 (pHydrion paper) by addition of 10% hydrochloric acid solution. The mixture was dissolved in 20 mL diethyl ether/water (1:1) and the organic layer was separated and sequentially washed with water and brine and then dried (Na ₂ S0 ₄ ) . Concentration afforded 2.03 g of an orange-red oil. The crude material was purified by flash column chromatography over silica gel using a solvent system of hexanes and ethyl acetate (90:10, R _f =0.23) . The yellow solid thus obtained was recrystallized from hexanes to afford a cream-colored, amorphous solid, mp 102-104°C. Spectral data: IR (Nujol) 3461, 1773 cm ^"1 ; ¹ H NMR: δ 1.45 (s, 3H) , 1.55 (s. 3H) , 3.95 (s. 2H) , 5.10 (S, IH) , 6.95 (s, IH) ; ¹³ C NMR: δ 30.10, 31.13, 34.49, 36.81, 107.50, 118.41, 133.69, 137.14, 148.77, 150.68, 174.82; HRMS: calcd. for C ₁₆ H ₂₂ 0 ₃ 262.1569, fnd. 262.1570. Anal.: calcd. for C ₁₆ H ₂₂ 0 ₃ C, 73.25, H, 8.45, fnd. C, 73.00, H, 8.40.

EXAMPLE 38 Synthesis of 2.3.5-Trimethyl-1.4-benzoquinone-l-mono-oxime

To a solution of 2,3,5-trimethyl-l,4-benzoquinone (70 g, 0.51 mol) of hot ethanol (200 mL) was added a warm solution of hydroxylamine hydrochloride (35 g, 0.50 mol) and cone, hydrochloric acid (5 mL) in water (100 mL) . The resulting red solution was stirred for 10 minutes at 60°C, stoppered and allowed to cool overnight in a refrigerator. The bright yellow precipitate was collected by vacuum filtration and dried to yield 63.9 g (83% yield) of crude product. Recrystallization from 50% aq. ethanol afforded the product as a yellow solid, mp 175°C (dec.) (lit.: Smith, L.I.; Schubert, W.M. J ^" . Am. Chem . Soc. 1948, 70 , 2656; mp 182°C). Spectral data: ¹ H NMR (CD ₃ SOCD ₃ ) : S 1.90 (s, 3H) , 1.93 (s, 3H) , 2.30 (s, 3H) , 7.55 (s, IH) , 12.95 (s.

1H, exchangeable with D ₂ 0) ; ¹³ C NMR (CD ₃ COCD ₃ ) : δ 11.49, 12.94, 16.09, 120.53, 132.77, 137.39, 140.72, 149.04, 186.13.

EXAMPLE 39 Synthesis of 4-Hydroxy-2.3.5-trimethylaniline (25a) The method of Smith and Schubert, J . Am . Chem . Soc .

1948, 70 , 2656, was followed. 2,3,5-Trimethyl-l,4-benzoquinone- 1-mono-oxime (50 g, 0.30 mol) was dissolved in aq. sodium hydroxide solution (49.5 g, 1.24 mol of NaOH in 720 mL of water) . The resulting intense red solution was cooled in an ice bath and stirred while sodium dithionite (126.1 g, 0.73 mol) was added. The red color faded and a beige precipitate formed. The mixture was stirred for 2 hours and allowed to come to ambient temperature. It was then cooled in ice and filtered. The crude 25a was pressed dry, rapidly transferred to an evacuated atmosphere and dried further (50°C/0.3 torr, 24 hours), mp 130-131°C with dec. (lit.: Smith, L.I.; Schubert, W.M. J . Am . Chem . Soc . 1948, 70 , 2656; 132-134°C dec) . Spectral data: ¹ H NMR (CD ₃ SOCD ₃ ) : δ 1.90 (s, 3H) , 2.05 (2s, 3H each), 4.10 (br s, 2H) , 6.25 (s, IH) , 7.10 (br s, IH) ; ¹³ C NMR (CD ₃ SOCD ₃ ) : δ 12.88, 13.25, 16.78, 114.39, 118.43, 122.25, 123.95, 138.81, 144.11.

EXAMPLE 40 Synthesis of 4-Hydroxy-2.3.5-trimethylpivalanilide (25b)

A solution of 4-hydroxy-2, 3,5-trimethylaniline (99.2 g, 0.66 mol) , pivaloyl chloride (87.1 g, 0.72 mol) and pyridine (1.4 L) was stirred for 16 hours under argon. The reaction mixture was poured onto ice and extracted into diethyl ether. The organic layer was sequentially washed with 10% hydrochloric acid, aq. sodium bicarbonate solution, water and brine. Drying and concentration afforded a purple solid. This solid was washed repeatedly with a solvent mixture of ethyl acetate and hexanes (3:7) to yield 91 g (59% yield) of the desired compound as a brown solid. Spectral data: IR (Nujol) 3410, 3250, 1652 cm ^'1 ; ¹ H NMR (CD ₃ SOCD ₃ ) : δ 1.35 (s, IH) , 2.10 (s, 3H) , 2.15 (ε, 3H) , 2.20 (s, 3H) , 5.35 (br s, IH) , 6.95 (s, IH) , 7.10 (br s, IH) .

EXAMPLE 41 Synthesis of 4-Acetoxy-2.3.5-trimethylpivalanilide (25c)

A solution of 25b (50.5 g, 0.21 mol), acetic anhydride (30 mL) , triethylamine (59 mL) and dry dichloromethane (370 mL) was stirred for 14 hours. The reaction mixture was concentrated, and the residual brown solid was washed with water and diethyl ether and dried (oil pump) to yield 54.9 g (93% yield) of 25c as a cream-colored solid. Spectral data: IR (Nujol) 3340, 1764, 1644 cm ^"1 ; ¹ H NMR (CD ₃ SOCD ₃ ) : δ 1.25 (s, 9H) . 2.00 (s, 6H) , 2.05 (S, 3H) , 2.35 (s, 3H) , 6.85 (s, IH) , 8.90 (s, IH) ; ¹³ C NMR (CD ₃ SOCD ₃ ) : δ 3.01, 4.25, 5.07, 10.20, 17.41, 116.54, 116.75, 118.61, 121.89, 123.88, 135.62, 156.65, 160.45.

EXAMPLE 42 Synthesis of 4-Acetoxy-6-bromo-2.3,5-trimethylpivalanilide (25d) A solution of 25c (5 g, 18 mmol) , bromine (1.85 mL, 36 mmol) and iodine (1.24 g, 4.9 mmol) in glacial acetic acid (65 mL) was heated between 65-73°C for 1 hour. The acetic acid was distilled (40°/O.3 torr) and the red viscouε residue, taken up in 75 mL of 1:1 CH ₂ Cl ₂ /hexanes, was washed with water, brine and dried (Na ₂ S0 ₄ ) . Concentration afforded a quantitative yield of 25d as an off-white solid. Spectral data: ¹ H NMR (CD ₃ SOCD ₃ ) : δ 1.25 (s, 9H) , 2.00 (s, 3H) , 2.10 (s, 3H) , 2.25 (s. 3H) , 2.45 (s, 3H) , 9.15 (s, IH) ; ¹³ C NMR (CD ₃ SOCD ₃ ) : δ 3.21, 5.30, 7.27, 10.19, 11.03, 17.34, 133.81, 117.62, 118.37, 123.51, 125.31, 136.25, 158.61, 165.96.

EXAMPLE 43 Synthesis of 6-Bromo-4-hydroxy-2.3.5-trimethylpivalanilide (2Se) By way of the same procedure that was used to deacetylate 23, crude 25d was isolated as a beige εolid (86% yield) by deacetylation of 25c. It was dried and used in the next step without further purification. Spectral data: ¹ H NMR: δ 1.25 (s, 9H) , 2.10 (ε, 3H) , 2.20 (ε, 3H) , 2.35 (s, 3H) , 3.65 (br s, >1H, [H ₂ 0]), 8.95 (s, IH) .

EXAMPLE 44 Synthesisof6-Bromo-4-tert-butyldimethylsiloxy-2.3,5-trimethy l- pivalanilide (25f)

Silylation of 25e was achieved by the method of Corey and Venkateswarlu, J. Am. Chem . Soc. 1972, 94 , 6190. The crude material was obtained in 91% yield. Spectral data: ¹ H NMR: *S

0.20 (S, 6H) , 1.00 (S, 9H) , 1.25 (s, 9H) , 2.05 (s, 3H) , 2.15 (ε,

3H) , 2.25 (S, 3H) , 8.95 (s, IH) .

EXAMPLE 45 Synthesis of iV-Allyl-6-bromo-4-tert-butyldimethylsiloxy-2.3.5- tri-met hylpivalanilide (26)

Anilide 25f (15 g, 35 mmol) in tetrahydrofuran (400 mL) was added dropwise over 1 hour to a εolution of allyl bromide (6 mL, 69 mmol) and previously washed and dried sodium hydride (1.68 g, 70 mmol) in tetrahydrofuran (50 mL) . The latter solution was maintained at -20°C during the addition process and subsequently allowed to approach ambient temperatures over 17 hours. It was quenched with ice water and the organic phase was then separated.

The aqueous phaεe waε waεhed with diethyl ether (3 x 75 mL) , and the combined organic layerε waε washed with brine and dried

(Na ₂ S0 ₄ ) . Concentration afforded quantitative yields of a mustard-colored oil. Purification of this oil by flash column chromatography on silica gel using a solvent εyεtem of hexaneε/ethyl acetate (9:1, R _f = 0.26) afforded 26 (88% yield) as a colorleεε, viεcouε oil. Spectral data: ¹ H NMR: δ 0.15 (s,

6H) , 0.95 (ε, 9H) , 1.05 (ε, 9H) , 2.15 (s, 3H) , 2.20 (s, 3H) , 2.30

(s, 3H) , 3.70-3.80 (dd, IH) , 4.50-4.60 (dd, IH) , 5.00 (m, 2H) ,

5.85-6.05 (m, IH) .

EXAMPLE 46 Synthesis of iV-Pivaloyl-5-hydroxy-3.4.6,7-tetramethylindole (27)

Indole 27 was prepared by the Pd-catalyzed cyelization of 26 according to the general procedure described earlier. The reaction temperature waε maintained at 100°C for 4.5-5 h and, after work-up, a waxy olive green residue was obtained. Purification waε achieved by flaεh column chromatography on silica gel using a solvent system of increasing polarity

(hexanes/ethyl acetate, 9:1, 8.5:1.5 and 8:2, R _f = 0.30 in the

last solvent system) . The pure material , an amorphous white solid, mp 151.5-153°C, was typically obtained in 40-45% yields. Spectral data: IR (Nujol) 3485, 1679, 1163 cm ^"1 ; ¹ H NMR:

(CD ₃ SOCD ₃ ) : δ 1.40 (s, 9H) , 1.90 (S, 3H) , 2.20 (s, 3H) , 2.35 (s, 3H) , 2.45 (s, 3H) , 7.40 (s, IH) , 7.85 (s, IH) ; ¹³ C NMR (CD ₃ SOCD3) : «S 1.97, 3.16, 3.40, 7.56, 18.72, 31.14, 103.84, 106.77, 111.43, 111.92, 114.19, 118.08, 122.11, 139.03, 168.33; CI-MS: m/z 274 (MH ⁺ , base) , 256, 189; CI-HRMS (M+H*) : calcd. for C ₁₇ H ₂₄ N0 ₂ 274.1807, fnd. 274.1792. Anal: calcd. for C ₁₇ H ₂₃ N0 ₂ C, 74.69, H, 8.48, N, 5.12, fnd. C, 74.81, H, 8.49, N, 5.14.

EXAMPLE 47 Synthesis of 4-Acetoxy-2.3.5-trimethylacetanilide (28a)

Anilide 28a was prepared in 80% yield by the diacetylation of 4-hydroxy-2, 3, 5-trimethylaniline according to the method of Gilbert and Pinto, J ^" . Org. Chem. 1992, 57, 5271- 5276. The product precipitated from the organic layer during work-up and was collected aε a white εolid by vacuum filtration. It was vacuum-dried and used in the next step without further purification. Spectral data: ¹ H NMR (CD ₃ SOCD ₃ ) : δ 2.00 (3 partially resolved s, 12H) , 2.35 (s, 3H) , 7.00 (s, IH) , 9.38 (s, IH) ; ¹³ C NMR (CD ₃ SOCD ₃ ) : δ 13.01, 14.37, 15.82, 20.19, 23.01, 125.51, 126.53, 128.72, 130.33, 133.63, 145.25, 168.25, 168.73.

EXAMPLE 48 Svntheεiε of 4-Acetoxy-6-bromo-2, 3 , 5-trimethylacetanilide (28b) A εolution of 28a (4.3 g, 18.2 mmol), bromine (1.8 mL,

36 mmol) and iron powder (0.27 g, 5 mmol) in glacial acetic acid (120 mL) waε heated at 80°C for 5 hours. The cooled reaction mixture was poured into ice-cold sodium bisulfite solution. The resulting fine white powder waε iεolated by vacuum filtration, washed with water and diethyl ether and dried at 60°C/0.2-0.5 torr. Anilide 28b waε obtained in 80% yield and uεed without further purification. Spectral data: ¹ H NMR (CD ₃ SOCD ₃ ) : δ 1.90 (ε, 3H) , 2.00 (ε, 3H) , 2.10 (s, 3H) , 2.20 (s, 3H) , 2.35 (s, 3H) , 9.55 (s, IH) ; ¹³ C NMR (CD ₃ SOCD ₃ ) : <S 13.21, 15.62, 17.29, 20.18, 22.46, 123.22, 127.67, 128.49, 133.28, 135.05, 146.28, 168.12, 168.64.

EXAMPLE 49

Synthesis of N-Allyl-4-acetoxγ-6-bromo-2.3.5-trimethylacetanilide '28c)

The alkylation of 28b was conducted in the same manner as for the alkylation of 25f but with the following changes: dimethyl formamide was used in place of tetrahydrofuran, a 10- to 15-fold excess of allyl bromide was employed, the reaction was initiated and carried out at room temperature and over a 3-hour reaction time and, the dimethyl formamide was distilled from the reaction mixture before quenching. Pure 28b was obtained aε a colorless oil (60% yield) after flash column chromatography over neutral alumina uεing a solvent system of Skellysolve B/ethyl acetate (7:3, R _f = 0.27) . Spectral data: 'H NMR (CD ₃ SOCD ₃ ) : δ 1.65 (s, 3H) , 2.05 (s, 3H) , 2.20 (ε, 3H) , 2.23 (ε, 3H) , 2.38 (s, 3H) , 3.80-3.90 (dd, IH) , 4.30-4.40 (dd, IH) , 5.10 (m, IH) , 5.80- 5.95 (m, IH) ; ¹³ C NMR (CD ₃ SOCD ₃ ) : δ 13.38, 16.09, 17.43, 20.18, 21.72, 50.64, 118.67, 124.00, 129.09, 129.87, 132.97, 135.75, 137.57, 147.17, 168.40, 169.14.

EXAMPLE 50 Synthesisof.V-Acetyl-5-acetoxy-3.4.6,7-tetramethylindoline (29a)

Cyelization of 28b and work-up of the reaction mixture was conducted analogously to that for the cyelization of 8. Pure

29a was obtained in 80% yield by purification of the crude material by flaεh column chromatography on silica gel using a solvent system of increasing polarity, namely, hexanes/ethyl acetate: 8:2, 6:4, 4:6, 2:8, R _f = 0.35 in the laεt syεtem. Spectral data: NMR (CD ₃ SOCD ₃ ) : δ 1.10 (d, 3H) , 1.95 and 1.96 (2 partially reεolved s, 6H and 3H respectively), 2.20 (br s, 3H) , 2.30 (s, 3H) , 3.15-3.25 (m, IH) , 3.75-3.90 (br m, IH) , 3.90-4.05 (br m, IH) ; ¹³ C NMR (CD ₃ SOCD ₃ , 90°C, 500 MHz) : δ 11.50, 12.17, 16.54, 17.48, 19.56, 22.57, 34.83, 58.06, 121.97, 124.86, 136.04, 138.37, 144.86, 168.01, 168.69.

EXAMPLE 51 Synthesis of 7V-Acetyl-5-hydroxy-3 ,4.6.7-tetramethylindoline (29b) Deacetylation of 29a was achieved by reaction with potassium carbonate in methanol according to the procedure of Burkalova, et al., Biofizika 1980, 24 , 989. Purification of

crude material by flash column chromatography on silica gel using a solvent system of ethyl acetate/Skellysolve B (6:4, R _f = 0.18) afforded 29b as a white solid, mp 171-173°C. Spectral data: IR (Nujol) 3500, 1635, 1400, 1210 cm ^"1 ; Η NMR (CD ₃ SOCD ₃ ) : δ 1.05 (d, 3H) , 1.90 (br s, 3H) , 2.10 (2 partially resolved s, 3H each), 2.15 (br 8, 3H) , 3.10 (br m, IH) , 3.75 (br m, IH) , 3.85 (br m, IH) , 7.85 (br s, IH) ; ¹³ C NMR (CD ₃ SOCD ₃ ) : δ 12.34, 12.64, 17.42, 18.10, 23.50, 35.44, 58.25, 117.49, 121.94, 124.93, 133.47, 135.81, 150.41, 168.58; CI-MS: m/z 234 (MH ⁺ , base), 172, 154, 136; CI-HRMS: Calcd. for C _u H _2rj N0 ₂ (M+H ⁺ ) 234.1494, fnd. 234.1494. Anal: calcd. for C ₁₄ H ₁₉ N0 ₂ C, 72.07, H, 8.21, N, 6.00, fnd. C, 71.85, H, 8.24, N, 5.92.

EXAMPLE 52 Synthesis of iV-(2-Methyl-2-propenyl) -4-acetoxy-6-bromo-2.3.5- trimethylpivalanilide (30)

Methallylation of 25d was accomplished analogously to the alkylation of 28b in dimethyl formamide. Flash column chromatography of the crude material on silica gel using a solvent syεtem of Skellyεolve B/ethyl acetate (60:40, R = 0.30) afforded a clear colorleεε oil (65% yield) . Spectral data: ¹ H NMR (CD ₃ SOCD ₃ ) : δ 1.65 (S, 3H) , 1.75 (S, 3H) , 2.05 (s, 3H) , 2.20 (S, 3H) , 2.25 (s, 3H) , 2.40 (s, 3H) , 3.65 (d, IH) , 4.35 (d, IH) , 4.60 (s, IH) , 4.70 (8, IH) .

EXAMPLE 53 Svntheεiε of iV-Pivaloyl-5-acetoxy-3.3.4.6,7-pentamethylindoline (31a)

Induline 31a waε prepared by the Pd-catalyzed cyelization of 30 according to the general procedure. The reaction was allowed to proceed at 85°C for 14 h. Purification of the crude olive green waxy residue by flash column chromatography on silica gel uεing a εolvent εyεtem of Skellyεolve B and ethyl acetate (6:4, R _f - 0.15) afforded a white solid (78% yield). Spectral data: ^] H NMR: δ 1.35 (br s, 6H) , 2.10 (ε, 3H) , 2.15 (ε, 6H) , 2.25 (br s, 3H) , 2.35 (s, 3H) , 3.70 (br s, 2H) ; CI-MS: m/z 290 (MH ⁺ , base), 247, 205, 190, 174.

EXAMPLE 54

Synthesis of N-Pivaloyl-5-hydroχy-3.3.4-6.7-pentamethylindoline

Olb)

Deacetylation of 31a was achieved as in the case of 29a. Product 31b was obtained as a white solid (79% yield), mp 164-165°C. Spectral data: H NMR (CD ₃ SOCD ₃ ) : <S 1.25 (s, 6H) , 1.90 (br s, 3H) , 2.10 (s, 3H) , 2.20 (ε, 3H) , 3.65 (br ε, 2H) , 7.85 (br ε, IH) ; ¹³ C NMR (CD ₃ SOCD ₃ ) : δ 11.85, 12.75, 17.24, 23.41, 24.68, 42.57, 64.86, 117.90, 122.01, 124.93, 133.86, 137.71, 150.64, 168.32; CI-MS: m/z 248 (MH ⁺ , baεe) , 230, 205, 190, 167; CI-HRMS (M+H ⁺ ) : Calcd. for C ₁₅ H ₂₂ N0 ₂ 248.1650, fnd. 248.1644. Anal: calcd. for C ₁₅ H ₂₁ N0 ₂ C, 72.7, H, 8.50, N, 5.70, fnd. C, 72.86, H, 8.46, N, 5.59.

EXAMPLE 55 Syntheεiε of Spiror2.3-dihydro-5-hvdroxy-4.6-7-trimethylbenzo- furan-2.1'-cyclopropane ] (32)

The phenol 32 was prepared according to the methodε of Gilbert and Pinto, J. Org. Chem. 1992, 57, 5271.

EXAMPLE 56 Synthesis of 5-Hvdroxy-2.4.6.7-tetramethylbenzofuran (33)

Phenol 33 was prepared by the reaction of Tebbe's reagentwith5-(tert-butyldimethylsiloxy)-4,6,7-trimethyl-2 (3H) - benzofu ranone according to the eεtabliεhed procedure of Gilbert and Pinto, J. Org. Chem. 1992, 57, 5271. The exo-methylene compound so obtained was isomerized to the benzofuran by pasεing the crude mixture through a column of εilica gel. The benzofuran was then recrystallized from hexanes and ethyl acetate (8:2) and sublimed at 50-90°C and at 0.51-0.5 torr, mp 136-137°C (lit.: Zahalka, H. A., et al., J. Org. Chem. 1988, 53, 3739-3745, 137.1- 138.9°C). Spectral data: H NMR: <S 2.25 (s, 3H) , 2.30 (s, 3H) , 2.40 (ε, 3H) , 2.42 (ε, 3H) , 4.40 (ε, IH) , 6.28 (ε, IH) ; ¹³ C NMR: δ 11.94, 12.03, 12.21, 14.16, 101.36, 110.18, 117.08, 118.21, 126.40, 147.23, 148.56, 154.41; EI-MS 190 (M ⁺ , base), 189, 175, 43; HRMS: Calcd. for C ₁₂ H _u 0 ₂ 190.0994, fnd. 190.0988. Materials And Media

Cellε were cultured in the following media: 1) MDA MB 435 cells were cultured in Minimum Essential Media (MEM-HyClone,

Logan, UT) supplemented with 5% heat inactivated fetal bovine serum (HyClone) , 2 mM glutamine (Sigma, St. Louis, MO) , 1 mM sodium pyruvate (Sigma) , IX MEM Non-Essential Amino Acidε (Sigma) , 2X MEM Vitamins (Sigma) , and 100 IU/mL penicillin/100 μg/mL streptomycin (HyClone) ; 2) MCF-7 McGuire cells were cultured in Minimum Essential Media (MEM-HyClone, Logan, UT) supplemented with 5% heat inactivated fetal bovine serum (HyClone) , 2 mM glutamine (Sigma, St. Louis, MO) , and 100 IU/mL penicillin/100 μg/mL streptomycin (HyClone) ; 3) HL-60 cells were cultured in Iεcoveε Modified Dulbecco's Media (IMDM-HyClone, Logan, UT) supplemented with 5% heat inactivated fetal bovine serum (HyClone) , 2 mM glutamine (Sigma, St. Louis, MO) , 1 mM sodium pyruvate (Sigma) , IX MEM Non-Essential Amino Acids (Sigma) , 2X MEM Vitamins (Sigma) , and 100 IU/mL penicillin/100 μg/mL streptomycin (HyClone) ; 2) MCF-7 McGuire cells were cultured in Minimum Esεential Media (MEM-HyClone, Logan, UT) supplemented with 5% heat inactivated fetal bovine εerum (HyClone) , 2 mM glutamine (Sigma, St. Louiε, MO) , and 100 IU/mL penicillin/100 μg/mL εtreptomycin (HyClone) ; 3) HL-60 cellε were cultured in Iεcoves Modified Dulbecco's Media (IMDM-HyClone, Logan, UT) εupplemented with 5% heat inactivated fetal bovine serum (HyClone) , 2 mM glutamine (Sigma, St. Louiε, MO) , and 100 IU/mL penicillin/100 μg/mL εtreptomycin (HyClone) . Solubility and Dilution of Phenolic Compounds All compounds were handled aε if they were light sensitive. All compounds were initially dissolved in absolute ethanol and subsequently diluted to a final concentration of 0.5% ethanol with the appropriate medium.

EXAMPLE 57 Bioassay

For the bioassay, all cells were used at 2.5 x 10 ⁵ /mL. Cells were treated with each of the compounds at concentrations of 20, 10, 5, 1, 0.1, and 0.01 μg/mL , and 200 μl of each treatment group were plated in quadruplicate in a 96 well culture plate (Corning, Corning NY) . Plates were done in duplicate, one plate to be used for viability testing and the other plate to examine ³ H-TdR uptake (DNA synthesis) .

Plates were cultured for 48 hours at 37°C, 5% C0 ₂ . Eight hours prior to the end of incubation, ³ H-TdR was added to one of the duplicate plates and incubation continued for 8 hours. The cells were then harvested (trypsinization waε required to harvest MDA MB 435 and MCF-7 McGuire) . At the end of the incubation, the cells were removed from the wells (of the duplicate plate - no radioisotope added) and viability checked by the Trypan Blue Exclusion method. Percent viability and percent suppression of each treatment group were calculated. EXAMPLE 58

Antiproliferative Effects

With respect to the data shown in the Tables below, the data is arranged from greatest percent inhibition of DNA synthesis to lowest. "TOXIC" refers to the fact that the compound at this concentration killed the cells or greatly lowered their viability. "NT" refers to "Not Tested". The cellε were cultured for 48 hourε, pulεed 8 hourε with tritiated thymidine, harveεted and counted. MDA MB 435 is the abbreviation for the Metastic Human Breast Cancer Cell Line. MCF-7 McGuire is the abbreviation for the Estrogen Responεive Human Breast Cancer Cell Line and HL-60 is the Human Promyelocytic Leukemia Cell Line.

Table I shows that compound 27 (MP2.8) significantly inhibited the proliferation of MB-435, MCF-7 and HL-60 cells at a dose of 5 μg/mL. Table II shows that compound 24 (MP2.1) significantly inhibited the proliferation of MB-435 and MCF-7 cells at 10 μg/mL and HL-60 cells at a dose of 20 μg/mL. Table III shows that compound 6 (KJA1) significantly inhibited the proliferation of MCF-7 and HL-60 cells at 10 μg/mL, and MB-435 cells at 5 μg/mL. Table IV shows that compound 13 (K-IPMP) inhibited the proliferation of MB-435 and HL-60 cells at 20 μg/mL. Table V showε that compound 12b (K-IPPA) significantly inhibited the proliferation of MB-435, MCF-7 and HL-60 cells at a dose of 5-20 μg/mL.

TABLE I INHIBITION (%) OF DNA SYNTHESIS BY COMPOUND 27 (MP 2.8) Concentration of Compounds (ug/mL )

Cell Type 20 10 5 1 0.1 0.01 MB-435 97 91 54 15 4 NT

MCF-7 TX 86 57 14 0 NT

HL-60 97 91 61 12 0 NT

TABLE II

INHIBITION (%) OF DNA SYNTHESIS BY COMPOUND 24 (MP 2.1) Concentration of Compoundε (ug/mL)

Cell Type 20 _. lj) 5 1 0.1 0.01

MB-435 97 85 29 5 13 NT

MCF-7 97 85 25 4 0 NT

HL-60 47 30 5 0 7 NT

TABLE III

INHIBITION (%) OF DNA SYNTHESIS BY COMPOUND 6 (KJA1)

Concentration of Compounds (ug/mL)

Cell Type 20 10 5 1 0.1 0.01

MB-435 TOXIC TOXIC 80 0 3 NT MCF-7 TOXIC 37 13 10 0 NT

HL-60 TOXIC 89 59 24 3 NT

TABLE IV

INHIBITION (%) OF DNA SYNTHESIS BY COMPOUND 13 (K-IPMP) Concentration of Compounds (ug/mL) Cell Type 20 10 5 1 _J. 0.01

MB-435 45 17 8 -4 4 NT

MCF-7 NT NT NT NT NT NT

HL-60 53 27 15 10 0 NT

TABfrg V INHIBITION (%) OF DNA SYNTHESIS BY COMPOUND 12b (K-IPPA)

Concentration of Compounds (u /mL )

Cell Type 20 10 5 1 0.1 0.01 MB-435 41 24 32 19 1 NT MCF-7 70 58 38 47 20 NT HL-60 46 67 32 2 0 NT

EXAMPLE 59 Table VI shows that compound 33 (MP2.6) significantly inhibited the proliferation of MB-435, MCF-7 and HL-60 cells at a dose of 20 μg/mL . Table VII shows that compound lib (K-IPP) significantly inhibited the proliferation of MB-435 cells at 10 μg/mL and MCF-7 cells at 5 μg/mL, while having virtually no effect on HL-60 cells. Table VIII showε that compound 6 (KJA1) significantly inhibited the proliferation of MB-435 and MCF-7 cells at 10-20 μg/mL, while having virtually no effect on HL-60 cells. Table IX shows that compound 32 (MP2.5) significantly inhibited the proliferation of MB-435, MCF-7 and HL-60 cells at a dose of 10-20 μg/mL. Table X shows that compound 20 (K-TBDP) inhibited the proliferation of MB-435 and HL-60 cells at 20 μg/mL; MCF-7 cellε were not teεted with thiε compound. Table XI shows that compound 31b (MP2.9) significantly inhibited the proliferation of MB-435, MCF-7 and HL-60 cells at a dose of 10-20 μg/mL. Table XII showε that compound 16 (K-ADMP) εignificantly inhibited the proliferation of MB-435 and HL-60 cellε at 5-20 μg/mL; MCF-7 cells were not tested with thiε compound.

TABLE VI

INHIBITION (%) OF DNA SYNTHESIS BY COMPOUND 33 (MP2.6) Concentration of Compoundε (μg/mL) Cell Type 20 JL0 5 1 0.1 0.01 MB-435 42 16 8 2 1 NT MCF-7 72 16 0 0 19 NT HL-60 36 7 0 0 0 NT

TABLE VII INHIBITION (%) OF DNA SYNTHESIS BY COMPOUND l b (K-IPP)

Concentration of Compoundε (ug/mL)

Cell Type 20 AP_ 5 1 0.1 0.01

MB-435 36 22 21 21 0 NT

MCF-7 53 33 47 26 24 NT

HL-60 0 0 0 0 0 NT

TABLE VIII

INHIBITION (%) OF DNA SYNTHESIS BY COMPOUND lie (K-TBP) Concentration of Compoundε (μg/mL) Cell Type 2_o 10 _. 5 l o.l o.oi

MB-435 31 12 20 12 13 NT

MCF-7 43 31 9 1 18 NT

HL-60 0 0 0 0 0 NT

TABLE IX

INHIBITION (%) OF DNA SYNTHESIS BY COMPOUND 32 (MP2.5) Concentration of Compounds (μg/mL)

Cell Type 2_o it) 5 l o.l o.oi

MB-435 43 30 15 6 6 NT MCF-7 22 18 5 10 0 NT

HL-60 26 17 12 5 11 NT

TABLE X

INHIBITION (%) OF DNA SYNTHESIS BY COMPOUND 20 (K-TBDP) Concentration of Compounds (μg/mL) Cell Type 20 .10 5 1 0^.1 0.01 MB-435 26 -13 -1 -5 -1 NT

MCF-7 NT NT NT NT NT NT

HL-60 63 33 18 6 2 NT

TABLE XI INHIBITION (%) OF DNA SYNTHESIS BY COMPOUND 31b (MP2.9) Concentration of Compounds (μg/mL) Cell TVPe 20 10 5 1 0_j_l 0.01 MB-435 33 15 0 0 0 NT

MCF-7 22 18 29 0 0 NT

HL-60 28 15 6 0 0 NT

TABLE XII

INHIBITION (%) OF DNA SYNTHESIS BY COMPOUND 16 (K-ADMP) Concentration of Compounds (μg/mL) Cell TVPe 20. 10 5 1 0.1 O.OI

MB-435 31 18 9 1 0 NT

MCF-7 NT NT NT NT NT NT

HL-60 15 11 11 0 0 NT

EXAMPLE 60

The present invention has potential for the efficacy of 5-hydroxyindoleε and 5-hydroxy-2, 3-dihydroindole derivativeε of the preεent invention as therapeutic agents by in vivo analyses of immune status and health of chickens inoculated with avian erythroblaεtoεiε viruε or avian erythroblaεtoεiε viruε- transformed tumor cells. Avian Erythroblaεtoεiε Viruε (AEV) Model:

The in vivo εtudieε utilize an avian retroviruε erythroleukemia model, namely, the avian erythroblastosis virus model. The avian erythroblastosis virus causes a rapid cachexia and fatal leukemia in 100% of challenged young birds. Newly hatched chickens are fed the 5-hydroxyindoles and 5-hydroxy-2, 3- dihydroindole derivativeε of the preεent invention supplemented diets or are intraperitoneally injected daily with the 5- hydroxyindoleε and 5-hydroxy-2, 3-dihydroindole derivativeε of the present invention for one week prior to tumor induction. For tumor induction, the chickens are injected with erythroleukemia cells or infectious avian erythroblastosiε viruε and their health (weight, immune εtatus, red blood cell status, e.g., anemia, and viability) are monitored.

EXAMPLE 61 Supplementation with 27 (MP2.8):

The maximum level of the 5-hydroxyindole derivative 27 (MP 2.8) that can be fed or injected without negatively affecting the health and growth of chickens is being established. These studies will also measure levels of 27 in the serum. Newly hatched SC strain White Leghorn chickens from Hy-line International (Dallas Center, IA) are to be used. Upon arrival the chicks will be assigned randomly (10/group/brooder) . Chicks will be housed in thermostatically controlled battery brooders with raised wire floors modified to prevent the loss of feed and are given a 15-hour day (15 hours light, 9 hours dark) .

The chickens will be observed daily and food consumption (brooder average values) will be determined every other day. Body weightε will be determined three timeε/week by weighing chickε. Blood (plaεma) will be taken at weekly intervalε, hematocritε will be performed and hexane extractε prepared and analyzed for the 5-hydroxyindole derivative 27, using high-performance liquid chromatography. At the conclusion of the study, chickens will be sacrificed by carbon dioxide anesthesia performed in compliance with recommendations of the Panel on Euthanasia of the American Veterinary Medical Association. Internal organs will be examined for any type of gross morphological evidence of 27 toxicity. The content of 27 in splenic bursal and thy ie lymphocyteε will be determined after density gradient purification and saponification and extraction into hexane by HPLC. These studies will permit the determination of levels of MP 2.8 that can be adminiεtered to young chickε, and determine the levelε of MP 2.8 found in plaεma, peripheral blood red blood cellε, splenic, bursal, and thymie lymphoid cells of chickens.

EXAMPLE 62 Analyses of health of chickens challenged with AEV

Upon arrival newly hatched chicks (10 chickens/group/brooder) will be assigned randomly to control groups and groups supplemented with 27. Eight days later (day 10 post hatch) , control chicks and thoεe εupplemented with 27

will be challenged with intravenous injections of 2.5 xlO ⁵ FFU AEV in 0.2 mL volume of phosphate buffered saline/animal or 1 x 10 ⁴ AEV tumor cells in 0.2 mL volume of phosphate buffered saline per animal. Previous studies have shown that this dose of virus or tumor cells when injected into chickens less than five weeks of age, induces fatal erythroleukemia involving weight loss, thymie atrophy and immune suppression in 100% of the chickens within 3 weeks. (Rao, A., Kline, K., and Sanders, B.G. Immune Abnormalities in Avian Erythroblastosis Virus-Infected Chickens. Cancer Research 50: 4764-4770, 1990) . The general health of the chickens will be observed daily, and body weight of individual chickens will be measured two times per week. Initially, tumor progression will be determined by comparing the time of death of virally and tumor injected chickens in non-27 supplemented controls versus 27-supplemented groups. The day of death is recorded aε the day a chicken dieε or is sacrificed in moribund condition.

Generally, 10-14 dayε following virus or tumor injection, all virus or tumor injected non-27 supplemented control chickens are dead. Supplementations with 27 will continue until control animals die (permitting evaluation of the ability of 27 to prolong life) or until an animal iε judged to be tumor free based on ability to survive 4 weeks beyond expected time of death with no histological evidence of erythroleukemia. At this time supplementation with 27 is terminated. These chickens are observed for general health, and body weights, and are assessed weekly for another four weeks when the experiment is terminated.

If supplementations with 27 diminish or prevent lethal erythroleukemia, additional experiments iε conducted to determine if 27 supplementation reduces cachexia (preventε weight loss, anemia, and immune suppreεsion) . Thus, 27 supplemented virus injected chickens and controls is used. Weights are measured every other day. Starting at 6 days post-viruε injection and every other day until day 14, five chickens from control and treatment groups are sacrificed by carbon dioxide anesthesia, bled for hematological analyses and spleen and thymus removed for

immune analyseε. The immune status is analyzed using standard T cell lectin-induced mitogenesiε and εtandard co-culture analyses for the presence of T suppressor cell activity.

Any patents or publications mentioned in this specification are indicative of the levels of thoεe skilled in the art to which the invention pertains. These patentε and publicationε are herein incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference. One skilled in the art will readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantageε mentioned, aε well aε those inherent therein. The present examples along with the methods, procedures, treatments, moleculeε, and εpecific compounds described herein are presently representative of preferred embodiments, are exemplary, and are not intended as limitations on the scope of the invention. Changeε therein and other uses will occur to those skilled in the art which are encompassed within the εpirit of the invention aε defined by the scope of the claims.