Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
  • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
  • C07D311/78—Ring systems having three or more relevant rings
  • C07D311/80—Dibenzopyrans; Hydrogenated dibenzopyrans
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents

Definitions

• the family of natural products known as the schweinfurthins includes four compounds isolated from the African plant Macaranga schweinfurthii Pax (see Beutler, J. A. et al., J. Nat. Prod. 1998, 61, 1509-1512; and Beutler, J. A., et al., Nat. Prod. Lett. 2000, 14, 349-404).
• Schweinfurthins A, B, and D display significant activity in the NCI's 60-cell line anticancer assay with mean GIso's ⁇ 1 ⁇ M. Their biological activity has attracted interest because some CNS, renal, and breast cancer cell lines are among the types most sensitive to these compounds. Inspection of the spectrum of activity shows no correlation with any currently used agents and suggests that these compounds may be acting at a previously unrecognized target or through a novel mechanism.
• schweinfurthins as cancer therapeutics would be accelerated by elucidating their mechanism of action.
• schweinfurthin analogues are fluorescent under UV light.
• SF-295 cells human glioblastoma multiforme
• 3-deoxy schweinfurthin B failed to generate a significant fluorescent signal above autofluorescence of control cells.
• the invention provides a compound of formula (I):
• R 1 and R 2 are each independently H, (Q-C ⁇ ⁇ lkyl, halo(C 1 -C 6 )alkyl or (C 3 - Cs)cycloalkyl; or one Of R 1 and R 2 is carboxy and the other is H, (Ci-C 6 )alkyl, ImIo(C 1 -C 6 )alkyl or (C 3 -Cg)cycloalkyl;
• R 3 is H, (C 1 -C 15 )alkyl, (C 2 -C 15 )alkenyl, (C 2 -C 15 )alkynyl, (C 1 -C 15 ) ⁇ kOXy, (C 1 - C 15 )alkylthio, (Ci-d 5 )alkanoyl, (Ci-C 15 )alkoxycarbonyl, (C 2 -C 15 )alkanoyloxy, hydroxy, mercapto, halo, cyano, or NR a R b ;
• R 4 is H, hydroxy, (C 1 -C 15 )alkyl, (C 2 -C 15 )alkenyl, (C 2 -C 15 )alkynyl, (C 1 -C 15 ) k 0 Xy, (C 1 - C!5)alkylthio, (Ci-C 15 )alkanoyl, (d-C 15 )alkoxycarbonyl, (C 2 -C 15 )alkanoyloxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, or NR c R d ; R 5 is aryl or heteroaryl, which aryl or heteroaryl is substituted with one or more groups
• R a and R b are each independently H, (C 1 -C 6 )alkyl, or (C 1 -C 15 )alkanoyl;
• R c and R d are each independently H, (C t -C ⁇ ⁇ lkyl, or (C 1 -Ci 5 )alkanoyl;
• the invention provides a therapeutic method for treating cancer comprising administering to a mammal in need of such therapy, an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof.
• the invention also provides a compound of formula (I) for use in medical therapy (e.g. for use in treating cancer), as well as the use of a compound of formula (I) for the manufacture of a medicament useful for the treatment of cancer in a mammal, such as a human.
• the invention also provides a compound of formula (I), or a pharmaceutically acceptable salt thereof for the prophylactic or therapeutic treatment of cancer.
• the invention also provides a method for identifing the sub-cellular localization of the target of the schweinfurthins comprising contacting cells with a fluoresent schweinfurthin analog (e.g. a compound of formula I or II) and detecting the location of the fluorescent compound in the cells in order to identify the sub-cellular localization of the target of the schweinfurthins.
• a fluoresent schweinfurthin analog e.g. a compound of formula I or II
• the invention also provides processes and intermediates disclosed herein that are useful for preparing compounds of formula (I) as well as other Schweinfurthin analogs.
• alkyl, alkoxy, alkenyl, alkynyl, etc. denote both straight and branched groups; but reference to an individual radical such as propyl embraces only the straight chain radical, a branched chain isomer such as isopropyl being specifically referred to.
• Alkenyl denotes a hydrocarbon chain with one or more (1, 2, 3, or 4) double bonds.
• alkynyl denotes a hydrocarbon chain with one or more (1, 2, 3, or 4) triple bonds.
• Aryl denotes a phenyl radical or an ortho-fused bicyclic carbocyclic radical having about nine to ten ring atoms in which at least one ring is aromatic; and heteroaryl encompasses a monocyclic aromatic ring containing five or six ring atoms consisting of carbon and one to four heteroatoms each selected from the group consisting of non-peroxide oxygen, sulfur, and N(X) wherein X is absent or is H, O, (Q-C ⁇ alkyl, phenyl or benzyl, as well as a radical of an ortho-fused bicyclic heterocycle of about eight to ten ring atoms derived therefrom, particularly a benz-derivative or one derived by fusing a propylene, trimethylene, or tetramethylene diradical thereto.
• enantiomerically enriched refers to mixtures that have one enantiomer present to a greater extent than another.
• the term “enantiomerically enriched” refers to a mixture having at least about 2% ee; in another embodiment of the invention, the term “enantiomerically enriched” refers to a mixture having at least about 5% ee; in another embodiment of the invention, the term “enantiomerically enriched” refers to a mixture having at least about 20% ee; in another embodiment of the invention, the term “enantiomerically enriched” refers to a mixture having at least about 50% ee; in another embodiment of the invention, the term “enantiomerically enriched” refers to a mixture having at least about 80% ee; in another embodiment of the invention, the term “enantiomerically enriched” refers to a mixture having at least about 90% ee; in another embodiment of the invention, the term “enantiomerically enriched”
• enantiomerically enriched includes enantiomerically pure mixtures which are mixtures that are substantially free of the species of the opposite optical activity or one enantiomer is present in very low quantities, for example, 0.01%, 0.001% or 0.0001%.
• protecting group or "blocking group” refers to any group which, when bound to a hydroxy prevents undesired reactions from occurring at this group and which can be removed by conventional chemical or enzymatic steps to reestablish the hydroxyl group.
• removable blocking group is not critical and preferred removable hydroxyl blocking groups include conventional substituents such as allyl, benzyl, acetyl, chloroacetyl, thiobenzyl, benzylidine, phenacyl, methyl methoxy, silyl ethers (e.g., t-butyl-diphenylsilyl or t- butylsilyl (“TBS”)) and any other group that can be introduced chemically onto a hydroxyl functionality and later selectively removed either by chemical or enzymatic methods in mild conditions compatible with the nature of the product.
• Suitable hydroxyl protecting groups are known to those skilled in the art and disclosed in more detail in T.W. Greene, Protecting Groups In Organic Synthesis; Wiley: New York, 1981, and the references cited therein.
• (Q-C ⁇ alkyl can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec- butyl, t-butyl, pentyl, 3-pentyl, hexyl, heptyl, octyl, nonyl, decyl, do-decyl, hexadecyl, octadecyl, icosyl;
• (C]-Ci 5 )alkoxy can be methoxy, ethoxy, propoxy, isopropoxy, butoxy, iso- butoxy, sec-butoxy, pentoxy, 3-pentoxy, or hexyloxy;
• (C 2 -C 15 )alkenyl can be vinyl, allyl, 1- propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1,-pentenyl, 2-pentenyl, 3-
• the invention provides a compound of formula (II):
• R 1 and R 2 are each independently H, (CrC 6 )alkyl, halo(C 1 -C 6 )alkyl or (C 3 - Cg)cycloalkyl; or one Of R 1 and R 2 is carboxy and the other is H, (Ci-C 6 )alkyl, halo(C 1 -C 6 )alkyl or (C 3 -C 8 )cycloalkyl; R 3 is H, (Ci-C 15 )alkyl, (C 2 -C 15 )alkenyl, (C 2 -C 15 )alkynyl, (C 1 - C 1S )EUCyItIIiO, (C 1 -C 15 )alkanoyl, (C 1 -C 15 )alkoxycarbonyl, (C 2 -C 15 )alkanoyloxy, hydroxy, mercapto, halo, cyano, or NR a R b ;
• R 4 is H, hydroxy, (C 1 -C 15 )alkyl, (C 2 -C 15 )alkenyl, (C 2 -C 15 )alkynyl, (C 1 - C 15 )alkylthio, (C 1 -C 15 )alkanoyl, (CrC ⁇ alkoxycarbonyl, (C 2 -C 15 )alkanoyloxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, or NR c R d ;
• R a and R b are each independently H, (CrC 6 )alkyl, or (Ci-C 15 )alkanoyl;
• R c and R d are each independently H, (d-C 6 )alkyl, or (CrC 15 )alkanoyl;
• R 1 is (C 1 -C 6 ) alkyl.
• R 1 is methyl
• R 2 is H.
• R 2 is (C 1 -C 6 ) alkyl. In one specific embodiment of the invention R 2 is methyl.
• R 3 is H.
• R 3 is hydroxy, amino, or mercapto.
• R 4 is H.
• R 4 is nitro. In one specific embodiment of the invention R 4 is hydroxy. In one specific embodiment of the invention R 4 is (Ci-C 1 S)EIkOXy.
• R 4 is methoxy
• R 5 is aryl that is substituted with one or two groups R x and that is also optionally substituted with one or more halo, hydroxy, cyano, trifluoromethyl, trifluoromethoxy, NR e R f , (C 2 -C 15 )alkenyl, (C 2 -C 15 )alkynyl, (CrCi 5 )alkoxy, (C 1 -C 15 )alkanoyl, (C 1 -C 15 )alkoxycarbonyl, or (C 2 -C 1 s)alkanoyloxy.
• R 5 is aryl that is substituted with one group R x and that is also optionally substituted with one or more halo, hydroxy, cyano, trifluoromethyl, trifluoromethoxy, NR e R f , (C 2 -C 15 )alkenyl, (C 2 -C 15 )alkynyl, (C r C 15 )alkoxy, (C 1 -C 15 )alkanoyl, (C 1 -C 1 5)alkoxycarbonyl, or (C 2 -C 15 )alkanoyloxy.
• R 5 is aryl that is substituted with one group R x and that is also optionally substituted with one or more halo, hydroxy, cyano, trifluoromethyl, trifluoromethoxy, NR e R f , (C 2 -C 15 )alkenyl, (C 2 -Ci 5 )alkynyl, (C 1 -C 15 )alkoxy, (Ci-C 15 )alkanoyl, (Ci-Ci 5 )alkoxycarbonyl, or (C 2 -C 15 )alkanoyloxy.
• R 5 is phenyl that is substituted with one or two groups R x and that is also optionally substituted with one or more halo, hydroxy, cyano, trifluoromethyl, trifluoromethoxy, NR e R f , (C 2 -C 15 )alkenyl, (C 2 -C 15 )alkynyl, (C 1 -C 15 )alkoxy, (C 1 -C 15 )alkanoyl, (C 1 -C 15 )alkoxycarbonyl, or (C 2 -Ci 5 )alkanoyloxy.
• R 5 is phenyl that is substituted with one group R x and that is also optionally substituted with one or more halo, hydroxy, cyano, trifluoromethyl, trifluoromethoxy, NR e R f , (C 2 -C 15 )alkenyl, (C 2 -C 15 )alkynyl, (CrCi 5 )alkoxy, (Ci-C 15 )alkanoyl, (Ci-C 15 )alkoxycarbonyl, or (C 2 -C 15 )alkanoyloxy.
• R 5 is phenyl that is substituted with one group R x and that is also optionally substituted with one or more halo, hydroxy, cyano, trifluoromethyl, trifluoromethoxy, NR e R f , (C 2 -C 15 )alkenyl, (C 2 -C i 5 )alkynyl, (Ci-C 15 )alkoxy, (Ci-C 15 )alkanoyl, (C 1 -C 15 )alkoxycarbonyl, or (C 2 -C 15 )alkanoyloxy.
• R 5 is of the formula
• R g and R k are each independently H, halo, hydroxy, (Q-Q ⁇ alkyl, (C 2 -C 15 )alkenyl, (C 2 -
• R 5 is of the formula
• R 8 and R k are each independently H, halo, hydroxy, (C 1 -Ci 5 )ahcyl, (C 2 -C 15 )alkenyl, (C 2 - Ci 5 )alkynyl, (CrC ⁇ alkoxy, methoxymethoxy, and (C 2 -C 15 )alkanoyloxy;
• R 8 and R k are each independently H, fluoro, chloro, bromo, hydroxy, or methoxy. In one specific embodiment of the invention R 8 and R k are each hydroxy.
• R 5 is heteroaryl that is substituted with one or two groups R x and that is also optionally substituted with one or more halo, hydroxy, cyano, trifluoromethyl, trifluoromethoxy, NR e R f , (C 2 -C 15 )alkenyl, (C 2 -C 15 )alkynyl, (Q-C ⁇ alkoxy, (C]-C 15 )alkanoyl, In one specific embodiment of the invention R 5 is heteroaryl that is substituted with one group R x and that is also optionally substituted with one or more halo, hydroxy, cyano, trifluoromethyl, trifluoromethoxy, NR e R f , (C 2 -C ⁇ alkenyl, (C 2 -C 1 s)alkynyl, (Ci-C ⁇ alkoxy, (C rC ⁇ alkanoyl, (C 1 -C 15 )alkoxycarbony
• R 5 is heteroaryl that is substituted with one group R x and that is also optionally substituted with one or more halo, hydroxy, cyano, trifluoromethyl, trifluoromethoxy, NR e R f , (C 2 -C 15 )alkenyl, (C 2 -C 15 )alkynyl, (Ci-C 15 )alkoxy, (C 1 -Ci 5 )alkanoyl, (Ci-Ci 5 )alkoxycarbonyl, or (C 2 -Ci 5 )alkanoyloxy.
• R 5 is an isoxazolyl, imadazolyl, pyridyl, indolyl, or benzo[b]furanyl ring that is substituted with one group R x and that is also optionally substituted with one or more halo, hydroxy, cyano, trifluoromethyl, trifluoromethoxy, NR e R f , (C 2 -Ci 5 )alkenyl, (C 2 -C i 5 )alkynyl, (Ci-Ci 5 )alkoxy, (Ci-C 15 )alkanoyl, (Ci-C 15 )alkoxycarbonyl, or (C 2 -C 15 )alkanoyloxy .
• each R x is independently R y .
• each R y is independently aryl, which is optionally substituted with one or more groups independently selected from hydroxy, (C 1 - C 6 )alkyl, (C 2 -C 6 )alkenyl, (C 2 -C 6 )alkynyl, (C 1 -C 6 )alkoxy, (C 1 -C 6 )alkylthio, (C 1 -C 6 )alkanoyl, (C 1 -C ⁇ )alkoxycarbonyl, (C 2 -C 6 )alkanoyloxy, halo, cyano, trifiuoromethyl, trifluoromethoxy, nitro, and NR V R W ; wherein each R v and R w is independently H, (C 1 -C 6 )alkyl, or (Ci- Ci 5 )alkanoyl.
• each R y is independently heteroaryl, which is optionally substituted with one or more groups independently selected from hydroxy, (C 1 - C 6 )alkyl, (C 2 -C 6 )alkenyl, (C 2 -C 6 )alkynyl, (C]-C 6 )alkoxy, (C 1 -C 6 )alkylthio, (Ci-C 6 )alkanoyl, (C 1 -C 6 )alkoxycarbonyl, (C 2 -C 6 )alkanoyloxy, halo, cyano, trifiuoromethyl, trifluoromethoxy, nitro, and NR V R W ; wherein each R v and R w is independently H, (C 1 -C 6 )alkyl, or (C 1 - Ci 5 )alkanoyl.
• each R y is independently phenyl, which is optionally substituted with one or more groups independently selected from hydroxy, (C 1 - C 6 )alkyl, (C 2 -C 6 )alkenyl, (C 2 -C 6 )alkynyl, (d-C 6 )alkoxy, (C r C 6 )alkylthio, (Ci-C 6 )alkanoyl, (C 2 -C 6 )alkanoyloxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, and NR V R W ; wherein each R v and R w is independently H, (Ci-C 6 )alkyl, or (Ci- Ci5)alkanoyl.
• each R y is independently phenyl which is optionally substituted with one or more groups independently selected from hydroxy, (Ci- C ⁇ jalkyl, (C 2 -C6)alkenyl, (Ci-C 6 )alkoxy, (Ci-C 6 )alkoxycarbonyl, (C 2 -C 6 )alkanoyloxy, halo, cyano, trifluoromethyl, trifluoromethoxy, nitro, and NR V R W ; wherein each R v and R w is independently H, (d-C 6 )alkyl, or (Ci-Ci 5 )alkanoyl.
• each R y is independently phenyl which is substituted with nitro or amino.
• the compound of formula (I) is selected from:
• the compound of formula (I) is selected from:
• the compound of formula (I) is isolated and purified.
• the invention provides a compound which is enantiomerically enriched and has an enantiomeric excess of at least about 90%.
• the invention provides a compound which is enantiomerically enriched and has an enantiomeric excess of at least about 95%.
• the invention provides a compound which is enantiomerically enriched and has an enantiomeric excess of at least about 98%.
• the invention provides a compound which is enantiomerically enriched and has an enantiomeric excess of at least about 99%. In one specific embodiment the invention provides a compound which is enantiomerically pure.
• the invention provides a compound of formula (I) which is the 2S 4aS 9aS enantiomer. In one specific embodiment the invention provides a compound of formula (I) which is the 2R 4aR 9aR enantiomer.
• salts are organic acid addition salts formed with acids which form a physiological acceptable anion, for example, tosylate, methanesulfonate, acetate, citrate, malonate, tartarate, succinate, benzoate, ascorbate, ⁇ -ketoglutarate, and ⁇ -glycerophosphate.
• Suitable inorganic salts may also be formed, including hydrochloride, sulfate, nitrate, bicarbonate, and carbonate salts.
• salts may be obtained using standard procedures well known in the art, for example by reacting a sufficiently basic compound such as an amine with a suitable acid affording a physiologically acceptable anion.
• a sufficiently basic compound such as an amine
• a suitable acid affording a physiologically acceptable anion.
• Alkali metal (for example, sodium, potassium or lithium) or alkaline earth metal (for example calcium) salts of carboxylic acids can also be made.
• Suitable acids includes any organic acid suitable to catalyze the reaction, such as, trifluoroacetic acid (TFA).
• Suitable base includes any base suitable to catalyze the reaction, such as, triethyl amine (TEA).
• the terms “isolated” and “purified” refer to substances that are substantially free of other biological agents, for example, at least about 95%, about 98%, or about 99% pure.
• the terms “treat,” “treatment,” and “treating,” extend to prophylaxis and include prevent, prevention, preventing, lowering, stopping or reversing the progression or severity of the condition or symptoms being treated. As such, the term “treatment” includes both medical, therapeutic, and/or prophylactic administration, as appropriate.
• compositions suitable for use in the present invention include those wherein the active compound is administered in an effective amount to achieve its intended purpose. More specifically, a "therapeutically effective amount” means an amount effective to treat the disease, disorder, and/or condition. Determination of a therapeutically effective amount is well within the capacity of persons skilled in the art, especially in light of the detailed disclosure provided herein.
• the pharmaceutically active compounds of the invention can be formulated as pharmaceutical compositions and administered to a mammalian host, such as a human patient in a variety of forms adapted to the chosen route of administration, e.g., orally or parenterally, by intravenous, intramuscular, topical or subcutaneous routes.
• the present compounds may be systemically administered, e.g., orally, in combination with a pharmaceutically acceptable vehicle such as an inert diluent or an assimilable edible carrier. They may be enclosed in hard or soft shell gelatin capsules, may be compressed into tablets, or may be incorporated directly with the food of the patient's diet.
• a pharmaceutically acceptable vehicle such as an inert diluent or an assimilable edible carrier.
• the active compound may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like.
• Such compositions and preparations should contain at least 0.1% of active compound.
• the percentage of the compositions and preparations may, of course, be varied and may conveniently be between about 2 to about 60% of the weight of a given unit dosage form.
• the amount of active compound in such therapeutically useful compositions is such that an effective dosage level will be obtained.
• the tablets, troches, pills, capsules, and the like may also contain the following: binders such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, fructose, lactose or aspartame or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring may be added.
• a liquid carrier such as a vegetable oil or a polyethylene glycol.
• any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed.
• the active compound may be incorporated into sustained-release preparations and devices.
• the active compound may also be administered intravenously or intraperitoneally by infusion or injection.
• Solutions of the active compound or its salts can be prepared in water, optionally mixed with a nontoxic surfactant.
• Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
• the pharmaceutical dosage forms suitable for injection or infusion can include sterile aqueous solutions or dispersions or sterile powders comprising the active ingredient which are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes.
• the ultimate dosage form for injection or infusion should be sterile, fluid and stable under the conditions of manufacture and storage.
• the liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof.
• the proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions or by the use of surfactants.
• the prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.
• Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filter sterilization.
• the preferred methods of preparation are vacuum drying and the freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the previously sterile-filtered solutions.
• the present compounds may be applied in pure form.
• compositions or formulations in combination with a dermatologically acceptable carrier, which may be a solid or a liquid.
• a dermatologically acceptable carrier which may be a solid or a liquid.
• Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silica, alumina and the like.
• Useful liquid carriers include water, alcohols or glycols or water-alcohol/glycol blends, in which the present compounds can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants.
• Adjuvants such as fragrances and additional antimicrobial agents can be added to optimize the properties for a given use.
• the resultant liquid compositions can be applied from absorbent pads, used to impregnate bandages and other dressings, or sprayed onto the affected area using pump-type or aerosol sprayers.
• Thickeners such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses or modified mineral materials can also be employed with liquid carriers to form spreadable pastes, gels, ointments, soaps, and the like, for application directly to the skin of the user.
• Examples of useful dermatological compositions which can be used to deliver the pharmaceutically active compounds of the invention to the skin are known to the art; for example, see Jacquet et al. (U.S. Pat. No. 4,608,392), Geria (U.S. Pat. No. 4,992,478), Smith et al. (U.S. Pat. No. 4,559,157) and Wortzman (U.S. Pat. No. 4,820,508).
• Useful dosages of the pharmaceutically active compounds of the invention can be determined by comparing their in vitro activity, and in vivo activity in animal models. Methods for the extrapolation of effective dosages in mice, and other animals, to humans are known to the art; for example, see U.S. Pat. No. 4,938,949.
• the amount of the compound, or an active salt or derivative thereof, required for use in treatment will vary not only with the particular salt selected but also with the route of administration, the nature of the condition being treated and the age and condition of the patient and will be ultimately at the discretion of the attendant physician or clinician.
• the compounds of the invention can also be administered in combination with other therapeutic agents that are effective to treat cancer.
• the desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day.
• the sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations; such as multiple inhalations from an insufflator or by application of a plurality of drops into the eye.
• R 1 -R 4 have any of the values or specific values defined herein; and wherein R 5a is a group of formula R 5 having any of the values or specific values defined herein or a group of formula R 5 that bears one or more protecting groups.
• the compound of formula (I) can be prepared by removing the protecting groups to provide the compound of formula (I).
• the corresponding compounds of formula (I) wherein the bond represented by — is a single bond can be prepared by reduction of the olefin with magnesium in methanol.
• the invention provides a method for preparing a compound of formula I comprising reacting an aldehyde of formula 20 or a corresponding aldehyde bearing one or more protecting groups, with a phosphonate of formula 21 or a corresponding phosphonate bearing one or more protecting groups, and optionally removing any protecting groups, to provide the compound of formula I.
• Intermediate aldehyde 20 is particularly useful for preparing compounds of formula I, and represents one specific embodiment of the invention.
• Halogen-lithium exchange of bromide 1 afforded the lithiated arene which was subsequently allowed to react with dry dimethylformamide affording the aldehyde 2 in acceptable yield.
• This aldehyde was treated with the known phosphonate 3 under modified Homer- Wadsworth-Emmons conditions giving the protected stilbene 4. Removal of the silyl protecting group under standard conditions gave access to the benzylic alcohol 5.
• a three step procedure was then used to convert the alcohol 5 into the benzylic phosphonate.
• treatment of the alcohol with methanesulfonyl chloride and triethyl amine affords the mesylate which can be smoothly transformed into the iodide.
• Another intermediate aldehyde that is useful for preparing compounds of formula I is a compound of formula 134:
• the compound of formula 134 can be prepared as illustrated below and as described in
• this aldehyde can be prepared from benzyl alcohol 115, which itself was available in 3 steps and 94% overall yield from vanillin. Methylation via a Williamson ether synthesis provided compound 116, which was then exposed to n- BuLi to induce halogen metal exchange. Reaction of the resulting aryl anion with geranyl bromide (17) furnished intermediate 118 in excellent overall yield. The methyl ether 118 was easily purified by column chromatography, which allowed preparation of this intermediate on a 5- to 10-gram scale.
• Ketone 124 then served as a platform for numerous oxidations.
• the Rubottom approach was explored using the more reactive silyl triflates to encourage enol ether formation. Even so, conversion to the silyl enol ether was incomplete (-60%), as observed by 1 H NMR analysis of a reaction conducted in CD 2 Cl 2 or by analysis of the initial product mixture. When this mixture was treated with mCPBA, the best result obtained was a 9% isolated yield of acyloin 125. Attempted MoOPH oxidation of ketone 124 afforded only recovered staring material as did the recent procedure of Tomkinson. When more forcing conditions were attempted with this oxidation, only decomposition was observed.
• Ketone 124 proved to be more reactive to oxidation by O 2 under basic conditions, but in this case the only isolated product had undergone rearrangement to an acid tentatively assigned structure 126, a product similar to one observed by Danishefsky, which may result from a Favorskii-like process.
• Ketone 131 was reduced upon treatment with NaBH 4 in quantitative yield to afford alcohol 133 as the only observed diastereomer.
• Exposure of compound 133 to DDQ afforded aldehyde 134 directly from the methyl ether.
• Compounds of formula I e.g. compounds 219, 225, 227, 235, and 237) can be prepared as illustrated below and as described in Examples 3-7.
• the nitro-stilbene 225 could in principle be used to identify the sub-cellular localization of the target of the schweinfurthins. In order for this to be truly informative it was determined that a fluorescent control compound lacking important pharmacophore elements would be highly useful. Reduction of the nitro group allowing entrance to amine analogues which could be used as a point of attachment to aldehyde decorated solid supports was also envisioned. These then could be used for chromatographic style isolation of interacting cellular components. Rounding out the set of desired compounds would then be the amine analogue of the control compound without the schweinfurthin warhead, and nitro and amine containing compounds based on the newly available schweinfurthin B A-ring diol system. These design priorities led to the identification of stilbenes 230, and 232 as fluorescent controls, and to stilbenes 227, and 237 as amine substituted targets.
• the target compounds all exhibited fluorescence (Table 1).
• the nitro-compounds 219, 225, 230, and 235 all displayed larger stokes shifts than the amine compounds 227, 232 and 237.
• the nitro substituted compounds also demonstrate significantly red-shifted absorbtion and emission maxima compared to the previously synthesized meta-phenol bis-stilbene 210. This shift to the green region for the fluorescence emission allows cellular studies on these compounds without any interference from autofluroescence.
• Table 1
• the anti-cancer activity of a compound of the invention may be determined using pharmacological models which are well known to the art, for example, NCI 60-cell line anticancer assay. Representative compounds of formula (I) were tested and were found to have anti-cancer activity in this assay.
• the effect of a compound of the invention on cancer cell morphology may be determined using pharmacological models which are well known to the art, or it may be determined using Test A below.
• Cell Culture-S ⁇ -295 cells were maintained in RPMI 1640 supplemented with 10% fetal calf serum, and penstreptamycin. A-549 cells were maintained in F- 12 media with 10% fetal calf serum and penstreptamycin. Both cell lines were incubated at 37° C and 5% CO .
• SF-295 cells are amongst the most sensitive cell lines to schweinfurthin induced growth inhibition. In addition to their effects on cellular growth, schweinfurthins induce drastic changes in cell morphology at time points beyond 24 hours.
• SF-295 cells are large polygonal shaped cells with multiple outstretched focal adhesions. Treatment of SF-295 cells with schweinfurthins such as 3-dSB, induces changes hallmarked by a decrease in cell area and a reduction in F-actin, which results in spindle-shaped cells with F-actin staining only at the periphery of these cells.
• the schweinfurthins display large differential, which means that some cell lines are less sensitive to schweinfurthins.
• One of these cell lines is the A-549 (human lung carcinoma) cell line.
• A-549 cells are rectangular and cover less surface area in comparison to SF-295 cells.
• Treatment with 3-dSB (50OnM) or compound 9 (50OnM) does not induce a morphologic change at 24 hours or 48 hours.
• the absence of change is consistent with previous studies which indicate that A549 cells are less sensitive to schweinfurthin treatment.
• Y-27632 (lO ⁇ M) induces ruffled F-actin at the edges of the A-549 cells.
• Selective toxicity is a distinguishing characteristic of the schweinfurthin compounds. Preserving the toxicity of the natural schweinfurthins is useful in the utilization of schweinfurthin analogues of formula I. Compound 9 maintains the activity of schweinfurthins in SF-295 cells and displays characteristic changes in cell morphology.
• the in vivo fluorescent properties of a compound of the invention may be determined using models which are well known to the art, or they may be determined using Test B below.
• Fluorescence Microscopy-SF -295 cells were plated on sterilized coverslips in 6-well plates and allowed to reach 65% confluency and then treated for indicated intervals. At the conclusion of the treatment interval cells were washed three times in complete media to remove residual compound. Immediately following rinsing cells were mounted onto microscope slides and imaged using a Bio-Rad Multi-photon microscope at the University of Iowa Central Microscopy facility. Images were further processed using Image-J software.
• Compounds of the invention can be used to elucidate the mechanism or schweinfurthin activity via compound localization. Indeed, SF-295 cells treated with concentrations of compound 9 as low as 10OnM could be visualized at all time points tested. Unlike previous attempts with other schweinfurthins, the fluorescence in treated cells was significantly greater than control cell fluorescence. As expected treatment of SF-295 cells with increasing concentrations of compound 9 increased fluorescence intensity at all time points tested. The localization of compound 9 within cells appears largely cytosolic with intense fluorescence in the peri-nuclear region. The fluorescent properties of compound 9 allow for its visualization which may allow for the determination of its site of action.
• the anti-cancer effect of a compound of the invention can also be determined using the assay scheme discussed in Test C below.
• the National Cancer Institute 60 human tumor cell line anti-cancer assay has been used for indicating the schweinfurthin-like activity of various analogues. Additionally, a three pronged approach that allows a more rapid turn around can be used. This three pronged testing scheme involves 1) MTT assay in schweinfurthin sensitive human glioma derived SF-295 cell line; 2) MTT assay in the schweinfurthin resistant human non-small cell lung cancer derived cell line A549; and 3) microscopic observation of cell morphology changes at 24 and 48 hours. Compounds displaying schweinfurthin-like activity show a dramatic change in cell morphology at concentrations consistent with anti-cancer activity.
• the invention provides a method for identifying a compound with schweinfurthin-like activity comprising, subjecting the compound to 1) an MTT assay in a schweinfurthin sensitive human glioma derived SF-295 cell line; 2) an MTT assay in the schweinfurthin resistant human non-small cell lung cancer derived cell line A549; and 3) a microscopic observation of cell morphology changes at one or more preselected time points (e.g. at about 24 or 48 hours).
• the nitro-substituted compound 225 and the control compound lacking the schweinfurthin left-half warhead 230 displayed activity as expected in this regard.
• the additional analogues synthesized here were also tested in this scheme.
• the free resorcinol 219 was the most potent of these compounds with almost identical activity to the standard 3- deoxyschweinfurthin B in the SF-295 cell line.
• 219 is almost an order of magnitude more potent than 3-deoxyschweinfurthin B in the A549 lung cancer line.
• This compound still shows the significant drop in activity in the A549 line vs. the SF-295 line that is indicative of the schweinfurthin like activity. All of the compounds showed an approximately 10 fold decrease in activity in the lung cancer vs.
• the intermediate stilbene 8 was prepared as follows.
• the reaction mixture was concentrated in vacuo to afford a red solid, which was dissolved in EtOAc. After the resulting yellow solution was washed with Na 3 S 2 O 3 until the color faded, it was washed with brine, dried (MgSO 4 ) and concentrated in vacuo. The resulting yellow oil was added to triethyl phosphite (2 mL) and the mixture was heated at 80 0 C for 24 h. After the solution was allowed to cool to rt, the excess phosphite was removed at high vacuum.
• the intermediate methyl ether 133 was prepared from the benzyl alcohol 115 as described below.
• Tricyclic Ether 121 To a solution of epoxide 120 (958 mg, 2.6 mmol) in CH 2 Cl 2 (350 mL) at -78 0 C was added BF 3 OEt 2 (2.0 mL, 16 mmol). After 7 min the reaction was quenched by addition of TEA (4.1 mL, 29 mmol). The resulting solution was concentrated in vacuo, dissolved in CH 2 Cl 2 , and washed with water then brine. The organic phase was dried (MgSO 4 ) and concentrated in vacuo.
• Ketone 124 To a solution of tricycle 121 (119 mg, 0.28 mmol) in CH 2 Cl 2 at room temperature was added TPAP (9 mg, 0.03 mmol) and NMO (49 mg, 0.41 mmol). After 18.5 hours the reaction mixture was diluted with ethyl acetate, filtered through celite, and concentrated in vacuo.
• Alcohol 129 To a solution of ketone 127 (1.75 g, 4.3 mmol) in CH 3 OH at room temperature was added CeCl 3 -7H 2 O (1.81 g, 4.9 mmol) followed by NaBH 4 (300 mg, 7.9 mmol). After 20 min, the reaction was quenched by addition of water and concentrated in vacuo. The resulting solution was extracted with ethyl acetate, and the combined extracts were washed with brine, dried (MgSO 4 ), and concentrated in vacuo, to afford alcohol 129 (1.75 g, 100%) as white crystals.
• Ketone 131 To a solution of compound 130 (35 mg, 0.08 mmol) in acetone was added NaHCO 3 (14 mg, 0.17 mmol) followed by KMnO 4 (23 mg, 0.15 mmol). After 20 hours at rt, additional NaHCO 3 (70 mg, 0.83 mmol) and KMnO 4 (20 mg, 0.13 mmol) was added. After an additional 24 hours at rt, the reaction mixture was filtered through celite, washed with acetone, and concentrated in vacuo.
• the intermediate protected stilbene 218 was prepared as follows
• the reaction mixture was concentrated in vacuo to afford a red solid, which was dissolved in EtOAc. After the resulting yellow solution was washed with Na 2 S 2 O 3 until the color faded, it was washed with brine, dried (MgSO 4 ) and concentrated in vacuo. The resulting yellow oil was added to triethyl phosphite (2 mL) and toluene (2 mL) the reaction mixture was heated at 110 0 C for 24 h. After the solution was allowed to cool to it, the excess phosphite was removed at high vacuum.
• the intermediate compound 224 was prepared as follows.
• the intermediate compound 223 was prepared as follows.
• the intermediate amine 226 was prepared as follows.
• the intermediate compound 234 was prepared as follows.
• a. Amine 236 To a solution of nitro compound 235 (16 mg, 0.024 mmol) in acetone (2 niL) was added sat. NH 4 Cl (1 mL) followed by Zn dust (26 mg, 0.4 mmol) and the mixture was heated to reflux. After 80 min, the solution was cooled and decanted into a separatory runnel. The resulting solution was extracted with ethyl acetate, and the combined organic phases were washed with brine.
• Example 8 Using procedures similar to those described herein, the following compounds of formula (I) were also prepared.
• Example 9 The following illustrate representative pharmaceutical dosage forms, containing a compound of formula (I) ('Compound X'), for therapeutic or prophylactic use in humans.

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