USRE41614E1 - Synthetic analogs of ecteinascidin-743 - Google Patents

Synthetic analogs of ecteinascidin-743 Download PDF

Info

Publication number
USRE41614E1
USRE41614E1 US10/738,973 US73897303A USRE41614E US RE41614 E1 USRE41614 E1 US RE41614E1 US 73897303 A US73897303 A US 73897303A US RE41614 E USRE41614 E US RE41614E
Authority
US
United States
Prior art keywords
group
compound
alkyl
ethyl acetate
halogen
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Lifetime
Application number
US10/738,973
Inventor
Elias J. Corey
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Harvard University
Original Assignee
Harvard University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Harvard University filed Critical Harvard University
Priority to US10/738,973 priority Critical patent/USRE41614E1/en
Application granted granted Critical
Publication of USRE41614E1 publication Critical patent/USRE41614E1/en
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/22—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains four or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • Et ecteinascidins
  • U.S. Pat. No. 5,721,362 which describes a synthetic process for the formation of ecteinascidin compounds and related structures, such as the saframycins.
  • the patent provides a synthetic route for the formation of ecteinascidin 743, an exceedingly potent marine-derived antitumor agent, now in clinical trials.
  • the process of this patent is enantio- and stereocontrolled, convergent and short.
  • novel process intermediates useful not only in the total synthesis of ecteinascidin 743, but also other known ecteinascidin compounds, including derivatives and analogs thereof.
  • Et 743 (NSC 648766) is currently undergoing evaluation by the National Cancer Institute on the basis of exceedingly potent activity in vivo against a variety of tumors.
  • This intermediate compound re-designated herein as Compound 1
  • the present invention is directed to compounds having the following formula:
  • Preferred compounds of the present invention have the following formula:
  • Suitable halogen substituents in the compounds of the present invention include F, Cl, Br and I.
  • Alkyl groups preferably have from 1 to about 12 carbon atoms, more preferably 1 to about 8 carbon atoms, still more preferably 1 to about 6 carbon atoms, and most preferably 1, 2, 3 or 4 carbon atoms.
  • Methyl, ethyl and propyl including isopropyl are particularly preferred alkyl groups in the compounds of the present invention.
  • the term alkyl unless otherwise modified, refers to both cyclic and noncyclic groups, although cyclic groups will comprise at least three carbon ring members.
  • alkenyl and alkynyl groups in the compounds of the present invention have one or more unsaturated linkages and from 2 to about 12 carbon atoms, more preferably 2 to about 8 carbon atoms, still more preferably 2 to about 6 carbon atoms, even more preferably 1, 2, 3 or 4 carbon atoms.
  • alkenyl and alkynyl as used herein refer to both cyclic and noncyclic groups, although straight or branched noncyclic groups are generally more preferred.
  • Preferred alkoxy groups in the compounds of the present invention include groups having one or more oxygen linkages and from 1 to about 12 carbon atoms, more preferably from 1 to about 8 carbon atoms, and still more preferably 1 to about 6 carbon atoms, and most preferably 1, 2, 3 or 4 carbon atoms.
  • Preferred alkylthio groups in the compounds of the present invention have one or more thioether linkages and from 1 to about 12 carbon atoms, more preferably from 1 to about 8 carbon atoms, and still more preferably 1 to about 6 carbon atoms. Alkylthio groups having 1, 2, 3 or 4 carbon atoms are particularly preferred.
  • Preferred alkylsulfinyl groups in the compounds of the present invention include those groups having one or more sulfoxide (SO) groups and from 1 to about 12 carbon atoms, more preferably from 1 to about 8 carbon atoms, and still more preferably 1 to about 6 carbon atoms.
  • Alkylsulfinyl groups having 1, 2, 3 or 4 carbon atoms are particularly preferred.
  • Preferred alkylsulfonyl groups in the compounds of the present invention include those groups having one or more sulfonyl (SO 2 ) groups and from 1 to about 12 carbon atoms, more preferably from 1 to about 8 carbon atoms, and still more preferably 1 to about 6 carbon atoms. Alkylsulfonyl groups having 1, 2, 3 or 4 carbon atoms are particularly preferred.
  • Preferred aminoalkyl groups include those groups having one or more primary, secondary and/or tertiary amine groups, and from 1 to about 12 carbon atoms, more preferably 1 to about 8 carbon atoms, still more preferably 1 to about 6 carbon atoms, even more preferably 1, 2, 3 or 4 carbon atoms.
  • Secondary and tertiary amine groups are generally more preferred than primary amine moieties.
  • Suitable heteroaromatic groups in the compounds of the present invention contain one, two or three heteroatoms selected from N, O or S atoms and include, e.g., coumarinyl including 8-coumarinyl, quinolinyl including 8-quinolinyl, pyridyl, pyrazinyl, pyrimidyl, furyl, pyrrolyl, thienyl, thiazolyl, oxazolyl, imidazolyl, indolyl, benzofuranyl and benzothiazol.
  • Suitable heteroalicyclic groups in the compounds of the present invention contain one, two or three heteroatoms selected from N, O or S atoms and include, e.g., tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, morpholino and pyrrolidinyl groups.
  • Suitable carbocyclic aryl groups in the compounds of the present invention include single and multiple ring compounds, including multiple ring compounds that contain separate and/or fused aryl groups.
  • Typical carbocyclic aryl groups contain 1 to 3 separate or fused rings and from 6 to about 18 carbon ring atoms.
  • carbocyclic aryl groups include phenyl including substituted phenyl, such as 2-substituted phenyl, 3-substituted phenyl, 2,3-substituted phenyl, 2,5-substituted phenyl, 2,3,5-substituted and 2,4,5-substituted phenyl, including where one or more of the phenyl substituents is an electron-withdrawing group such as halogen, cyano, nitro, alkanoyl, sulfinyl, sulfonyl and the like; naphthyl including 1-naphthyl and 2-naphthyl; biphenyl; phenanthryl; and anthracyl.
  • substituted phenyl such as 2-substituted phenyl, 3-substituted phenyl, 2,3-substituted phenyl, 2,5-substituted phenyl,
  • references herein to substituted R′ groups in the compounds of the present invention refer to the specified moiety that may be substituted at one or more available positions by one or more suitable groups, e.g., halogen such as fluoro, chloro, bromo and iodo; cyano; hydroxyl; nitro; azido; alkanoyl such as a C 1 -C 6 alkanoyl group such as acyl and the like; carboxamido; allyl groups including those groups having 1 3 to about 12 carbon atoms or from 1 3 to about 6 carbon atoms and more preferably 1-3 3 carbon atoms; alkenyl and alkynyl groups including groups having one or more unsaturated linkages and from 2 to about 12 carbon atoms or from 2 to about 6 carbon atoms; alkoxy groups having those having one or more oxygen linkages and from 1 to about 12 carbon atoms or 1 to about 6 carbon atoms; aryloxy such as phenoxy; alkylthio groups including those moi
  • the compounds of the present invention can be prepared synthetically from the intermediate compound 11 described in the '362 patent. Numerous active antitumor compounds have been prepared from this compound and it is believed that many more compounds may be formed in accordance with the teachings of the present disclosure.
  • One especially preferred embodiment of the present invention is the novel ecteinascidin-like compounds that have been prepared from Compound 1:
  • compositions useful as antitumor agents comprising an effective antitumor amount of one or more of the compounds of the present invention and a pharmaceutically acceptable diluent, carrier or excipient.
  • Yet another especially preferred embodiment of the present invention is the synthetic intermediates of the compounds of the present invention as described in detail below.
  • the present invention includes the synthetic processes described herein.
  • the currently most preferred compound of the present invention is the compound of formula 7:
  • the first step for producing the preferred compound 7 of the present invention is the high yield conversion (93%) of the phenol compound 1 to the allyl ether compound 2.
  • the second step is the high yield (99%) removal of the TBDPS protecting group to form the free alcohol compound 3.
  • the third step in this process is the high yield (91%) coupling of phthalimide to the free alcohol compound 3 to yield the phthalimide derivative, compound 4.
  • the phthalimide compound 4 is then converted in high yield (97%) to the phenol compound 5.
  • Phenol compound 5 is converted in high yield (94%) to the methoxymethyl ether compound 6.
  • the phthalimide compound 4 can be treated with several reagents to produce in high yield (91%) the methoxymethyl ether compound 6.
  • the methoxymethyl ether compound 6 is finally reacted with trifluoroacetic acid to provide the desired compound 7, in high yield (94%).
  • the overall yield of this process is about 72%.
  • the phenol compound 5 may be transformed into a number of derivatives, as shown in Scheme III:
  • reaction schemes described herein may be modified and/or combined in various ways, and the compounds generated therefrom are to be considered as being part of this invention.
  • Alcohol (3) (61.5 mg, 0.109 mmol) and phthalimide (18.8 mg, 0.128 mmol) were azeotropically dried with toluene (2 ⁇ 5 mL) and dissolved in THF (3.8 mL).
  • Triphenylphosphine (35.0 mg, 0.133 mmol) was added followed by diethyl azodicarboxylate (19.0 ⁇ L, 0.121 mmol). The reaction turned yellow and then a bright orange color within 5 minutes. After stirring at 23° C. for 2 h the reaction was concentrated in vacuo at 23° C.
  • Phthalimide (4) (20.0 mg, 0.0289 mmol) and acetic acid (16.5 ⁇ L, 0.289 mmol) were dissolved in methylene chloride (0.8 mL).
  • PdCl 2 (PPh 3 ) 2 (1.0 mg, 1.4 ⁇ mol) was added followed by tributyltin hydride (21.0 ⁇ L, 0.0779 mmol). Bubbling was observed and the reaction changes from a yellow to a dark orange color. After stirring at 23° C. for 10 min the reaction was quenched into water (20 mL), extracted with methylene chloride (2 ⁇ 20 mL), dried over sodium sulfate, decanted and concentrated in vacuo.
  • Phenol (5) (1.1 mg, 0.0017 mmol) was dissolved in methylene chloride (0.15 mL). 4-Dimethylaminopyridine (0.5 mg, 0.0041 mmol) and acetic anhydride (0.5 ⁇ L, 0.0053 mmol) were added to the solution which was stirred at 23° C. for 30 min. The reaction mixture was concentrated in vacuo. The residue was purified by flash column chromatography (0.5 mL silica gel, gradient 1:4 to 1:1 to 2:1 ethyl acetate-hexane) to afford Compound 6 (1.1 mg, 94%).
  • phthalimide (4) (68.5 mg, 0.99 mmol) and acetic acid (17.0 ⁇ L, 0.30 mmol) were dissolved in methylene chloride (6.0 mL).
  • PdCl 2 (PPh 3 ) 2 (3.5 mg, 5 ⁇ mol) was added followed by tributyltin hydride (67.0 ⁇ L, 0.25 mmol). Bubbling was observed and the reaction changes from a yellow to a dark orange color.
  • triethylamine (55.0 ⁇ L, 0.40 mmol)
  • 4-dimethylaminopyridine 5.5 mg, 0.045 mmol
  • acetic anhydride (38.0 ⁇ L, 0.39 mmol).
  • the methoxymethyl ether (6) (3.8 mg, 0.00547 mmol) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 4.0 mL) and the solution was stirred at 23° C. for 7 h.
  • the reaction mixture was diluted with toluene (5 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3 ⁇ 5 mL).
  • phthalimides and dicarboximides were not commercially available and had to be synthesized from the commercially available anhydrides or dicarboxylic acids using a variety of established methodologies.
  • the dicarboxylic acids were converted to the anhydrides by heating with acetic anhydride. Heating the anhydrides with urea 1 , urethane 2 or formamide 3 at
  • the 1,2-Naphthalimide was synthesized via a Diels-Alder with ⁇ -bromostyrene and maleimide. 5 p-Toluenesulfonyl isocyanate and t-butanol were reacted in order to generate the BOC-protected tolylsulfonamide. 6 The dicarboximides were systematically dried under vacuum (60° C., 30 mm) and by toluene azeotrope immediately before use.
  • the alcohol (9) (1.0 mg, 0.0018 mmol) and the dicarboximide (0.0065 mmol, 3.6 equiv.) were azeotropically dried with toluene (2 ⁇ 0.1 mL) and dissolved in THF (0.2 mL).
  • Triphenylphosphine (1.7 mg, 0.0065 mmol) was added as a solid followed by diethyl azodicarboxylate (1.0 ⁇ L, 0.0064 mmol) via syringe.
  • the reaction turned yellow and after stirring at 23° C. 7 for 15 h the reaction was concentrated in vacuo.
  • the methoxymethyl ether was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.2 mL) and the solution was stirred at 23° C. for 11 h.
  • the reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3 ⁇ 1 mL).
  • the residue was purified by flash column chromatography (0.5 mL silica gel, gradient methylene chloride to 2:1-ethyl acetate-hexane) to afford the desired product.
  • Nitro compound (14) (0.5 mg, 0.00072 mmol) was dissolved in methanol (0.4 mL), 10% Pd/C (0.2 mg) and ammonium formate (12.0 mg, 0.19 mmol) were added at 23° C. and the reaction was stirred for 40 min. The mixture was diluted with ethyl acetate (2 mL), filtered through a plug of Celite, concentrated in vacuo and the residue was purified by flash column chromatography (1.5 mL silica gel, 2:1 ethyl acetate-hexane) to afford Compound 20 (0.3 mg, 63%).
  • Nitro compound (18) (0.5 mg, 0.00067 mmol) was dissolved in methanol (0.4 mL), 10% Pd/C (0.2 mg) and ammonium formate (12.0 mg, 0.19 mmol) were added at 23° C. and the reaction was stirred for 40 min. The mixture was diluted with ethyl acetate (2 mL), filtered through a plug of Celite, concentrated in vacuo and the residue was purified by flash column chromatography (1.5 mL silica gel, 2:1 ethyl acetate-hexane) to afford Compound 21 (0.4 mg, 83%).
  • Alcohol (9) (1.0 mg, 0.0018 mmol) was dissolved in methylene chloride (0.2 mL) and 4-dimethylaminopyridine (0.1 mg, 0.00082 mmol) and phenyl isocyanate (0.5 ⁇ L, 0.0046 mmol) were added to the solution.
  • the reaction was stirred at 23° C. for 3 hr and then quenched into a saturated solution of aqueous sodium bicarbonate (10 mL).
  • the mixture was extracted with methylene chloride (2 ⁇ 5 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo to afford a residue (1.2 mg, 100%).
  • Phthalimide (7) (0.3 mg, 0.00046 mmol) was dissolved in methylene chloride (0.2 mL) and 4-dimethylaminopyridine (0.6 mg, 0.0049 mmol) and acetic anhydride (1.0 ⁇ L, 0.010 mmol) were added to the solution. The reaction was stirred at 23° C. for 20 min and then purified by flash column chromatography (0.3 mL silica gel, gradient methylene chloride to ethyl acetate) to afford Compound 23 (0.3 mg, 94%).
  • Phthalimide (7) (0.7 mg, 0.0011 mmol) was dissolved in methylene chloride (0.2 mL) and N,N-diisopropylethylamine (1.0 ⁇ L, 0.0058 mmol) and N-chlorosuccinimide (0.66 mg, 0.0049 mmol) were added to the solution.
  • the reaction was stirred at 23° C. for 28 hr and passed through a small plug of silica gel with ethyl acetate. The mixture was concentrated in vacuo and the residue was purified by preparative thin layer chromatography (10% ethyl acetate-methylene chloride, three elutions) to afford Compound 24 (0.5 mg, 68%).
  • Phthalimide (7) (0.5 mg, 0.00077 mmol) was dissolved in a 0.0056 M solution of N-bromosuccinimide in methylene chloride (0.14 mL, 0.00079 mmol). The reaction was stirred at 23° C. for 40 min and was then quenched into a saturated solution of sodium thiosulfate (10 mL). The mixture was extracted with ethyl acetate (10 mL) and the organic layers were washed with water (2 ⁇ 20 mL) and saturated aqueous sodium chloride (20 mL), dried over sodium sulfate, decanted and concentrated in vacuo.
  • Phthalimide (7) (0.5 mg, 0.00077 mmol) was dissolved in 3:2 acetonitrile-water (0.25 mL). Silver nitrate (4.0 mg, 0.024 mmol) was added as a solid and the solution was stirred at 23° C. for 11 hr. The reaction was quenched by stirring with a 1:1 mixture of saturated aqueous sodium chloride and saturated aqueous sodium bicarbonate (0.5 mL) for 15 min.
  • Phthalimide (4) (3.6 mg, 0.0052 mmol) was azeotropically dried with toluene (2 ⁇ 2 mL) and dissolved in THF (0.5 mL). The mixture was cooled to ⁇ 78° C. in a dry ice-acetone bath and a 1.0 M solution of L-Selectride in THF (10 ⁇ L, 0.010 mmol) was added drop-wise. The reaction was warmed to 23° C. slowly over 5 hr and was quenched with 2 drops of 5% acetic acid in water. After stirring at 23° C.
  • Phthalimide (4) (3.6 mg, 0.0052 mmol) was azeotropically dried with toluene (2 ⁇ 2 mL) and dissolved in THF (0.5 mL). The mixture was cooled to ⁇ 78° C. in a dry ice-acetone bath and a 1.0 M solution of L-Selectride in THF (10 ⁇ L, 0.010 mmol) was added dropwise. The reaction was warmed to 23° C. slowly over 5 hr and was quenched with 2 drops of 5% acetic acid in water. After stirring at 23° C.
  • This material (3.0 mg, 0.004 mmol) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.2 mL) and the solution was stirred at 23° C. for 11 h.
  • the reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3 ⁇ 1 mL).
  • Alcohol (4) (14.3 mg, 0.025 mmol) was azeotropically dried with toluene (2 ⁇ 1 mL) in vacuo. The residue was dissolved in methylene chloride (0.5 mL) and to this solution was added N,N-diisopropylethylamine (9.0 ⁇ L, 0.052 mmol), 4-dimethylaminopyridine (9.4 mg, 0.077 mmol) and p-toluenesulfonic anhydride (29.0 mg, 0.089 mmol). The reaction was stirred at 23° C. for 13 hr and was then quenched into a half-saturated solution of aqueous sodium bicarbonate (10 mL).
  • Tosylate (29) (14.0 mg, 0.020 mmol) was dissolved in DMF (0.5 mL). Lithium azide (7.7 mg, 0.16 mmol) was added and the reaction was placed in a 70° C. oil bath for 20 min. The reaction was cooled to room temperature, diluted with 1:1 ethyl acetate-hexane (20 mL) and washed with water (3 ⁇ 20 mL) and saturated aqueous sodium chloride (20 mL). The organic layers were dried over sodium sulfate, decanted and concentrated in vacuo.
  • Tosylate (29) (1.0 mg, 0.0014 mmol) was dissolved in a saturated solution of potassium 4-pyridinedicarboimide (0.2 mL, ⁇ 30 equiv.). After stirring at 23° C. for 4 hr the reaction was diluted with 1:1 ethyl acetate-hexane (10 mL) and washed with water (10 mL). The aqueous was extracted with 1:1 ethyl acetate-hexane (10 mL) and the combined organic layers were washed with saturated aqueous sodium chloride (10 mL), dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by flash column chromatography (1.0 mL silica gel, gradient methylene chloride to 2:1 diethyl ether-hexane) to afford a residue (0.9 mg, 94%).
  • This material was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.2 mL) and the solution was stirred at 23° C. for 11 hr.
  • the reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3 ⁇ 1 mL).
  • Amine (31) (0.6 mg, 0.0011 mmol) was dissolved in methylene chloride (0.5 mL). To this mixture was added 4-dimethylaminopyridine (0.5 mg, 0.0041 mmol) and ⁇ , ⁇ -dibromoxylene (0.5 mg, 0.0019 mmol). After stirring at 23° C. for 3 hr the reaction was purified by flash column chromatography (0.6 mL silica gel, gradient methylene chloride to 1:1 ethyl acetate-hexane) to afford a film (0.5 mg, 71%).
  • Amine (31) (0.6 mg, 0.0011 mmol) was dissolved in methylene chloride (0.5 mL). To this mixture was added 4-dimethylaminopyridine (0.5 mg, 0.0041 mmol), pyruvic acid (0.5 ⁇ L, 0.0072 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.5 mg, 0.0026 mmol). After stirring at 23° C. for 3 hr the reaction was purified by flash column chromatography (0.6 mL silica gel, gradient methylene chloride to 1:1 ethyl acetate-hexane) to afford a film (0.5 mg, 73%).
  • Phenol (5) (1.0 mg, 0.0015 mmol) was dissolved in a 0.0126 M solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine in methylene chloride (0.5 mL, 0.0064 mmol of each).
  • the carboxylic acid was added and the reaction was stirred at 23° C. for 30 min and then quenched into a saturated solution of aqueous sodium bicarbonate (2 mL).
  • the mixture was extracted with methylene chloride (2 ⁇ 5 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo.
  • the residue was purified by flash column chromatography (1 mL silica gel, gradient methylene chloride to 1:1 to 2:1 ethyl acetate-hexane) to afford the corresponding phenolic esters.
  • This material was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL) and the solution was stirred at 23° C. for 11 h.
  • the reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3 ⁇ 1 mL).
  • the residue was purified by flash column chromatography (1 mL silica gel, gradient methylene chloride to 2:1 ethyl acetate-hexane to ethyl acetate) 8 to afford the desired product.
  • Entry 5 was purified using 5% methanol-methylene chloride as the eluent.
  • Phenol (5) (1.0 mg, 0.0015 mmol) was azeotropically dried with toluene (2 ⁇ 1 mL) in vacuo and dissolved in DMF (0.1 mL).
  • Cesium carbonate (3.0 mg, 0.0092 mmol) was gently flame dried in vacuo, cooled and added as a solid to the reaction mixture.
  • the alkylation agent was added via syringe and the solution was stirred at 23° C. for 4 hr and then quenched into a saturated solution of aqueous sodium bicarbonate (2 mL).
  • Phenol (5) (1.0 mg, 0.0015 mmol) was dissolved in methylene chloride (0.5 mL) to this solution were added 4-dimethylaminopyridine (0.8 mg, 0.0066 mmol) and n-butyric anhydride (1.0 ⁇ L, 0.0061 mmol). The reaction was stirred at 23° C. for 15 min and then quenched into a saturated solution of aqueous sodium bicarbonate (2 mL). The mixture was extracted with methylene chloride (2 ⁇ 5 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo.
  • Phenol (5) (1.0 mg, 0.0015 mmol) was dissolved in methylene chloride (0.5 mL) to this solution were added 4-dimethylaminopyridine (0.8 mg, 0.0066 mmol) and methanesulfonyl chloride (0.5 ⁇ L, 0.0065 mmol). The reaction was stirred at 23° C. for 15 min and then quenched into a saturated solution of aqueous sodium bicarbonate (2 mL). The mixture was extracted with methylene chloride (2 ⁇ 5 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo.
  • Methoxymethyl ether (4) (0.5 mg, 0.00072 mmol) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL) and the solution was stirred at 23° C. for 11 h.
  • the reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3 ⁇ 1 mL).
  • the residue was purified by flash column chromatography (0.5 mL silica gel, gradient methylene chloride to 2:1 ethyl acetate-hexane) to afford Compound 45 (0.4 mg, 85%).
  • the methoxymethyl ether (5) (5.0 mg, 0.0077 mmol) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.2 mL) and the solution. was stirred at 23° C. for 11 h under an atmosphere of oxygen. The reaction mixture was diluted with toluene (5 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3 ⁇ 5 mL). The residue was purified by preparative TLC (2:1 ethyl acetate-hexane) to afford Compound 46 (2.3 mg, 50%).
  • hydroxyquinone (46) (2.3 mg, 0.0038 mmol) was dissolved in methylene chloride (3 mL). A dilute diazomethane solution in diethyl ether was added in small portions while monitoring the reaction by TLC analysis. Upon complete conversion to the product, acetic acid (50 ⁇ L) was added to quench the reaction. Purification via preparative TLC (1:1 ethyl acetate-hexane) afforded pure (47) (1.0 mg, 42%).
  • the methoxymethyl ether (5) (0.6 mg, 0.00092 mmol) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL)) and the solution was stirred at 23° C. for 7 h.
  • the reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3 ⁇ 1 mL).
  • the residue was purified by flash column chromatography (0.4 mL silica gel, gradient methylene chloride to 2:1 ethyl acetate-hexane) to afford Compound 48 (0.4 mg, 71%).
  • Phthalimide (5) (5.4 mg, 0.0083 mmol) was dissolved in ethanol (0.3 mL) and hydrazine (26 ⁇ L, 0.829 mmol) was added. The vessel was sealed and heated to 80° C. for 2 h. The reaction was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (2 ⁇ 1 mL). The residue was purified by flash column chromatography (0.5 mL silica gel, gradient methylene chloride to 5% methanol-ethyl acetate) to afford Compound 49 (4.3 mg, 100%).
  • the methoxymethyl ether (50) (1.3 mg, 0.0019 mmol) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL) and the solution was stirred at 23 ⁇ C for 10 h.
  • the reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3 ⁇ 1 mL).
  • the residue was purified by flash column chromatography (0.3 mL silica gel, gradient 2:1 ethyl acetate-hexane to ethyl acetate) to afford Compound 51 (0.5 mg, 42%).
  • p-Hydroxyphenylacetic acid (100 mg, 0.657 mmol) was dissolved in DMF (3.0 mL).
  • tert-Butyldimethylsily chloride 222 mg, 1.47 mmol
  • N,N-diisopropylethylamine 0.285 mL, 1.64 mmol were added to the solution which was stirred at 23° C. for 3 h.
  • Water (1 mL) was added and after 15 min the reaction mixture was poured into 5% aqueous acetic acid (25 mL) and extracted with ethyl acetate (2 ⁇ 25 mL).
  • Amine (49) (2.0 mg, 0.0038 mmol) and acid (53) (1.3 mg, 0.0049 mmol) were azeotropically dried with toluene (2 ⁇ 1 mL) and then dissolved in methylene chloride (0.2 mL).
  • 1,3-Dicyclohexylcarbodiimide (1.0 mg, 0.0049 mmol) was added to the solution which was stirred at 23° C. for 30 min. White precipitate was observed and the reaction was quenched into saturated solution of aqueous sodium bicarbonate (5 mL). The aqueous layer was extracted with methylene chloride (2 ⁇ 5 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo.
  • Phenol (54) (1.1 mg, 0.0015 mmol) was dissolved in methylene chloride (0.2 mL). 4-Dimethylaminopyridine (0.4 mg, 0.0032 mmol) and acetic anhydride (0.5 ⁇ L, 0.0053 mmol) were added to the solution which was stirred at 23° C. for 1 h. The reaction mixture was concentrated in vacuo. The residue was purified by flash column chromatography (0.3 mL silica gel, gradient 1:1 ethyl acetate-hexane to ethyl acetate) to afford Compound 55 (1.2 mg, 100%).
  • the methoxymethyl ether (55) (1.2 mg, 0.0015 mmol) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL) and the solution was stirred at 23° C. for 10 h.
  • the reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3 ⁇ 1 mL).
  • the residue was purified by flash column chromatography (0.3 mL silica gel, ethyl acetate) to afford Compound 56 (0.4 mg, 44%).
  • the methoxymethyl ether (65) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 v/v), 1.0 mL) and the solution was stirred at 23° C. for 11 h.
  • the reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3 ⁇ 1 mL).
  • the residue was purified by preparative thin layer chromatography (2:1 diethyl ether-hexane, two elutions and 1:1 ethyl acetate-hexane) to afford Compound 63 (0.4 mg, 61%).
  • Phenol (5) (0.8 mg, 0.0012 mmol) was dissolved in THF (0.2 mL) and to this solution were added 4-dimethylaminopyridine (1.0 mg, 0.0082 mmol) and methylisocyanate (0.5 ⁇ L, 0.0085 mmol). The reaction was stirred at 23° C. for 19 h and then quenched into a saturated solution of aqueous sodium bicarbonate (2 mL). The mixture was extracted with 1:1 ethyl acetate-hexane (2 ⁇ 5 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo.
  • the analogs described above were screened in vitro for anti-tumor activity.
  • the human cancer cell lines used in these assays include A-549 (Lung), HCT116 (Colon), A375 (Melanoma) and PC-3 (Prostate) and values are reported as IC 50 (ng/mL).
  • IC 50 ng/mL
  • the following tables summarize the activity of all the synthetic derivatives. An IC 50 reading greater than 100 ng/mL is considered inactive in the screening tests conducted on the compounds of the present invention. Lower values represent higher activity.
  • the compounds of the present invention will serve as useful antitumor agents in mammals, particularly in humans.
  • Antitumor compounds are typically administered in unit dosage form.
  • Each unit dose refers to a physically discrete unit suitable as unitary dosages for animals, each unit containing a predetermined quantity of active material calculated to produce the desired antitumor effect in association with the required diluent; i.e., carrier, or vehicle.
  • the specifications for the novel unit dose of this invention are dictated by and are directly dependent on (a) the unique characteristics of the active material and the particular antitumor effect to be achieved, and (b) the limitations inherent in the art of compounding such active material for such use in mammals, particularly humans, as disclosed in detail herein, these being features of the present invention.
  • Unit dosage forms are typically prepared from the active compound by dispersement thereof in a physiologically tolerable (or acceptable) diluent or vehicle such as water, saline or phosphate-buffered saline, to form an aqueous composition. If necessary, other pharmaceutically acceptable solvents may be used.
  • physiologically tolerable (or acceptable) diluent or vehicle such as water, saline or phosphate-buffered saline.
  • other pharmaceutically acceptable solvents may be used.
  • Such diluents are well known in the art and are discussed, for example, in Remington's Pharmaceutical Sciences, 16th Ed., Mack Publishing Company, Easton, Pa. (1980) at pages 1465-1467.
  • Dosage forms can also include an adjuvant as part of the diluent.
  • adjuvants such as complete Freund's adjuvant (CFA), incomplete Fruend's adjuvant (IFA) and alum are materials well known in the art, and are available commercially from several sources.
  • the quantity of active compound to be administered depends, inter alia, on the animal species to be treated, the subject animal's size, the size of the tumor being treated (if known), and the capacity of the subject to utilize the active compound. Precise amounts of the active compound required to be administered depend on the judgment of the practitioner and are peculiar to each individual, particularly where humans are the treated animals. Dosage ranges, however, can be characterized by a therapeutically effective blood concentration and can range from a concentration of the active compound of the present invention from about 0.01 ⁇ M to about 100 ⁇ M, preferably about 0.1 ⁇ M to 10 ⁇ M.
  • Suitable regimes for initial administration and booster injections are also variable, but are typified by an initial administration followed by repeated doses at one or more hour intervals by a subsequent injection or other administration.
  • continuous intravenous infusion sufficient to maintain therapeutically effective concentrations in the blood are contemplated.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Public Health (AREA)
  • General Chemical & Material Sciences (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Plural Heterocyclic Compounds (AREA)
  • Saccharide Compounds (AREA)

Abstract

The present invention is directed to the synthesis and characterization of compounds having the formula:
Figure USRE041614-20100831-C00001
    • wherein R1, R2, R3, R4, R5, R6, R7, R8 and R9 are each independently selected from the group consisting of H, OH, OR′, SH, SR′, SOR′, SO2R′, NO2, NH2, NHR′, N(R′)2, NHC(═O)R′ NHC(═O)R′, CN, halogen, ═O, C(═O)H, C(═O)R′, CO2H, CO2R′, C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl, and substituted or unsubstituted heteroaromatic;
    • wherein each of the R′ groups is independently selected from the group consisting of H, OH, NH2, NO2, SH, CN, halogen, ═O, C(═O)H, C(═O)CH3, CO2H, CO2CH3, C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, aryl, aralkyl, and heteroaromatic;
    • wherein each dotted circle represents one, two or three optional double bonds;
    • wherein R7 and R8 may be are joined into a carbocyclic or heterocyclic ring system; and
    • wherein X1 and X2 are each independently defined as above for R1-R8 R1-R 6 and R 9, and each further includes specific preferred groups as defined herein.

Description

CROSS-REFERENCE TO RELATED APPLICATION
This application is a reissue of U.S. Pat. No. 6,348,467 and issued Feb. 19, 2002. The '467 patent was issued from U.S. application Ser. No. 09/510,315, filed Feb. 22, 2000 as a continuation of application U.S. Ser. No. 09/165,892, filed Sep. 30, 1998, now U.S. Pat. No. 6,124,292, the disclosure of which is hereby incorporated herein by reference.
STATEMENT OF GOVERNMENT SUPPORT
This invention was supported in part by funding from the National Institutes of Health under Grant No. R01 GM 34167 and the National Science Foundation under Grant Nos. CHE 9300276 and CHE 9811917. Accordingly, the government of the United States may have certain rights in this invention.
BACKGROUND OF THE INVENTION
The ecteinascidins (herein abbreviated Et or Et's) are exceedingly potent antitumor agents isolated from the marine tunicate Ecteinascidia turbinata. Several ecteinascidins have been reported previously in the patent and scientific literature. See, for example:
U.S. Pat. No. 5,721,362, which describes a synthetic process for the formation of ecteinascidin compounds and related structures, such as the saframycins. In one particularly preferred embodiment, the patent provides a synthetic route for the formation of ecteinascidin 743, an exceedingly potent marine-derived antitumor agent, now in clinical trials. The process of this patent is enantio- and stereocontrolled, convergent and short. Also disclosed are novel process intermediates, useful not only in the total synthesis of ecteinascidin 743, but also other known ecteinascidin compounds, including derivatives and analogs thereof.
U.S. Pat. No. 5,256,663, which describes pharmaceutical compositions comprising matter extracted from the tropical marine invertebrate, Ecteinascidia turbinata, and designated therein as ecteinascidins, and the use of such compositions as antibacterial, anti-viral, and/or antitumor agents in mammals.
U.S. Pat. No. 5,089,273, which describes novel compositions of matter extracted from the tropical marine invertebrate, Ecteinascidia turbinata, and designated therein as ecteinascidins 729, 743, 745, 759A, 759B and 770. These compounds are useful as antibacterial and/or antitumor agents in mammals.
U.S. Pat. No. 5,478,932, which describes ecteinascidins isolated from the Caribbean tunicate Ecteinascidia turbinata, which provide in vivo protection against P388 lymphoma, B16 melanoma, M5076 ovarian sarcoma, Lewis lung carcinoma, and the LX-1 human lung and MX-1 human mammary carcinoma zenografts.
U.S. Pat. No. 5,654,426, which describes several ecteinascidins isolated from the Caribbean tunicate Ecteinascidia turbinata, which provide in vivo protection against P388 lymphoma, B16 melanoma, M5076 ovarian sarcoma, Lewis lung carcinoma, and the LX-1 human lung and MX-1 human mammary carcinoma zenografts.
See also: Corey, E. J., J. Am. Chem. Soc., 1996, 118 pp. 9202-9203; Rinehart, et al., Journal of National Products, 1990, “Bioactive Compounds from Aquatic and Terrestrial Sources”, vol. 53, pp. 771-792; Rinehart et al., Pure and Appl. Chem., 1990, “Biologically active natural products”, vol. 62, pp. 1277-1280; Rinehart, et al., J. Org. Chem., 1990, “Ecteinascidins 729, 743, 745, 759A, 759B, and 770: Potent Antitumor Agents from the Caribbean Tunicate Ecteinascidia turbinata”, vol. 55, pp. 4512-4515; Wright et al., J. Org. Chem., 1990, “Antitumor Tetrahydroisoquinoline Alkaloids from the Colonial Ascidian Ecteinascidia turbinata”, vol. 55, pp. 4508-4512; Sakai et al., Proc. Natl. Acad. Sci. USA 1992, “Additional antitumor ecteinascidins from a Caribbean tunicate: Crystal structures and activities in vivo”, vol. 89, 11456-11460; Science 1994, “Chemical Prospectors Scour the Seas for Promising Drugs”, vol. 266, pp. 1324; Koenig, K. E., “Asymmetric Synthesis,” ed. Morrison, Academic Press, Inc., Orlando, Fla., vol. 5, 1985, p. 71; Barton, et al., J. Chem Soc. Perkin Trans., 1, 1982, “Synthesis and Properties of a Series of Sterically Hindered Guandidine Bases”, pp. 2085; Fukuyama et al., J. Am Chem Soc., 1982, “Stereocontrolled Total Synthesis of (+)-Saframycin B”, vol. 104, pp. 4957; Fukuyama et al., J. Am Chem Soc., 1990, “Total Synthesis of (+)-Saframycin A”, vol. 112, p. 3712; Saito, et al., J. Org. Chem., 1989, “Synthesis of Saframycins. Preparation of a Key Tricyclic Lactam Intermediate to Saframycin A”, vol. 54, 5391; Still, et al., J. Org. Chem., 1978, “Rapid Chromatographic Technique for Preparative Separations with Moderate Resolution”, vol. 43, p. 2923; Kofron, W. G.; Baclawski, L. M., J. Org. Chem., 1976, vol. 41, 1879; Guan et al., J. Biomolec. Struc. & Dynam., vol. 10 pp. 793-817 (1993); Shamma et al., “Carbon-13 NMR Shift Assignments of Amines and Alkaloids,” p. 206 (1979); Lown et al., Biochemistry, 21, 419-428 (1982); Zmijewski et al., Chem. Biol. Interactions, 52, 361-375 (1985); Ito, CRC Crit. Rev. Anal. Chem., 17, 65-143 (1986); Rinehart et al., “Topics in Pharmaceutical Sciences 1989” pp. 613-626, D. D. Breimer, D. J. A. Cromwelin, K. K. Midha, Eds., Amsterdam Medical Press B. V., Noordwijk, The Netherlands (1989); Rinehart et al., “Biological Mass Spectrometry,” 233-258 eds. Burlingame et al., Elsevier Amsterdam (1990); Guan et al., Jour. Biomolec. Struc & Dynam., vol. 10 pp. 793-817 (1993); Nakagawa et al., J. Amer. Chem. Soc., 111: 2721-2722 (1989); Lichter et al., “Food and Drugs from the Sea Proceedings” (1972), Marine Technology Society, Washington, D.C. 1973, 117-127; Sakai et al., J. Amer. Chem. Soc., 1996, 118, 9017; Garcia—Rocha et al., Brit. J. Cancer, 1996, 73: 875-883; and Pommier et al., Biochemistry, 1996, 35: 13303-13309.
The disclosures of the above-referenced patents and publications are hereby incorporated herein by reference.
Et 743 (NSC 648766) is currently undergoing evaluation by the National Cancer Institute on the basis of exceedingly potent activity in vivo against a variety of tumors.
Figure USRE041614-20100831-C00002
In 1996, the total synthesis of Et-743 was reported. See E. J. Corey et al., J. Amer. Chem. Soc., 118, 9292-9203 (1996); see also, U.S. Pat. No. 5,721,362. Disclosed in the '362 patent is the intermediate 11, with the following structure:
Figure USRE041614-20100831-C00003
This intermediate compound, re-designated herein as Compound 1, has served as the starting material for a series of new synthetic ecteinascidin-like compounds.
SUMMARY OF THE INVENTION
The present invention is directed to compounds having the following formula:
Figure USRE041614-20100831-C00004
    • wherein the substituent groups defined by R1, R2, R3, R4, R5, R6, R7, R8 and R9 are each independently selected from the group consisting of H, OH, OR′, SH, SR′, SOR′, SO2R′, NO2, NH2, NHR′, N(R′)2, NHC(O)R′, CN, halogen, ═O, C(═O)H, C(═O)R′, CO2H, CO2R′, C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl, and substituted or unsubstituted heteroaromatic;
    • wherein each of the R′ groups is independently selected from the group consisting of H, OH, NO2, NH2, SH, CN, halogen, ═O, C(═O)H, C(═O)CH3, CO2H, CO2CH3, C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, aryl, aralkyl, and heteroaromatic;
    • wherein each dotted circle represents one, two or three optional double bonds;
    • wherein R7 and R8 may be joined into a carbocyclic or heterocyclic ring system;
    • and wherein X1 and X2 are each independently defined as above for R1-R8 and further include the definitions of X1 and X2 as provided below for the preferred embodiments.
Preferred compounds of the present invention have the following formula:
Figure USRE041614-20100831-C00005
    • wherein the substituent groups defined by R1, R2, R3, R4, R5, R6, and R9 are each independently selected from the group consisting of H, OH, OR′, SH, SR′, SOR′, SO2R′, NO2, NH2, NHR′, N(R′)2, NHC(O)R′ NHC(═O)R′, CN, halogen, ═O, C1-C6 alkyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl, and substituted or unsubstituted heteroaromatic;
    • wherein each of the R′ groups is independently selected from the group consisting of H, OH, NO2, NH2, SH, CN, halogen, ═O, C(═O)H, C(═O)CH3, CO2H, CO2CH3, C1-C6 alkyl, phenyl, benzyl, and heteroaromatic;
    • wherein each dotted circle represents one, two or three optional double bonds;
    • and wherein X1 and X2 are each independently defined as above for R1-R8 R1-R 6 and R 9, and further include the definitions of X1 and X2 as provided below for the preferred embodiments.
Suitable halogen substituents in the compounds of the present invention include F, Cl, Br and I.
Alkyl groups preferably have from 1 to about 12 carbon atoms, more preferably 1 to about 8 carbon atoms, still more preferably 1 to about 6 carbon atoms, and most preferably 1, 2, 3 or 4 carbon atoms. Methyl, ethyl and propyl including isopropyl are particularly preferred alkyl groups in the compounds of the present invention. As used herein, the term alkyl, unless otherwise modified, refers to both cyclic and noncyclic groups, although cyclic groups will comprise at least three carbon ring members.
Preferred alkenyl and alkynyl groups in the compounds of the present invention have one or more unsaturated linkages and from 2 to about 12 carbon atoms, more preferably 2 to about 8 carbon atoms, still more preferably 2 to about 6 carbon atoms, even more preferably 1, 2, 3 or 4 carbon atoms. The terms alkenyl and alkynyl as used herein refer to both cyclic and noncyclic groups, although straight or branched noncyclic groups are generally more preferred.
Preferred alkoxy groups in the compounds of the present invention include groups having one or more oxygen linkages and from 1 to about 12 carbon atoms, more preferably from 1 to about 8 carbon atoms, and still more preferably 1 to about 6 carbon atoms, and most preferably 1, 2, 3 or 4 carbon atoms.
Preferred alkylthio groups in the compounds of the present invention have one or more thioether linkages and from 1 to about 12 carbon atoms, more preferably from 1 to about 8 carbon atoms, and still more preferably 1 to about 6 carbon atoms. Alkylthio groups having 1, 2, 3 or 4 carbon atoms are particularly preferred.
Preferred alkylsulfinyl groups in the compounds of the present invention include those groups having one or more sulfoxide (SO) groups and from 1 to about 12 carbon atoms, more preferably from 1 to about 8 carbon atoms, and still more preferably 1 to about 6 carbon atoms. Alkylsulfinyl groups having 1, 2, 3 or 4 carbon atoms are particularly preferred.
Preferred alkylsulfonyl groups in the compounds of the present invention include those groups having one or more sulfonyl (SO2) groups and from 1 to about 12 carbon atoms, more preferably from 1 to about 8 carbon atoms, and still more preferably 1 to about 6 carbon atoms. Alkylsulfonyl groups having 1, 2, 3 or 4 carbon atoms are particularly preferred.
Preferred aminoalkyl groups include those groups having one or more primary, secondary and/or tertiary amine groups, and from 1 to about 12 carbon atoms, more preferably 1 to about 8 carbon atoms, still more preferably 1 to about 6 carbon atoms, even more preferably 1, 2, 3 or 4 carbon atoms. Secondary and tertiary amine groups are generally more preferred than primary amine moieties.
Suitable heteroaromatic groups in the compounds of the present invention contain one, two or three heteroatoms selected from N, O or S atoms and include, e.g., coumarinyl including 8-coumarinyl, quinolinyl including 8-quinolinyl, pyridyl, pyrazinyl, pyrimidyl, furyl, pyrrolyl, thienyl, thiazolyl, oxazolyl, imidazolyl, indolyl, benzofuranyl and benzothiazol. Suitable heteroalicyclic groups in the compounds of the present invention contain one, two or three heteroatoms selected from N, O or S atoms and include, e.g., tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, morpholino and pyrrolidinyl groups.
Suitable carbocyclic aryl groups in the compounds of the present invention include single and multiple ring compounds, including multiple ring compounds that contain separate and/or fused aryl groups. Typical carbocyclic aryl groups contain 1 to 3 separate or fused rings and from 6 to about 18 carbon ring atoms. Specifically preferred carbocyclic aryl groups include phenyl including substituted phenyl, such as 2-substituted phenyl, 3-substituted phenyl, 2,3-substituted phenyl, 2,5-substituted phenyl, 2,3,5-substituted and 2,4,5-substituted phenyl, including where one or more of the phenyl substituents is an electron-withdrawing group such as halogen, cyano, nitro, alkanoyl, sulfinyl, sulfonyl and the like; naphthyl including 1-naphthyl and 2-naphthyl; biphenyl; phenanthryl; and anthracyl.
References herein to substituted R′ groups in the compounds of the present invention refer to the specified moiety that may be substituted at one or more available positions by one or more suitable groups, e.g., halogen such as fluoro, chloro, bromo and iodo; cyano; hydroxyl; nitro; azido; alkanoyl such as a C1-C6 alkanoyl group such as acyl and the like; carboxamido; allyl groups including those groups having 1 3 to about 12 carbon atoms or from 1 3 to about 6 carbon atoms and more preferably 1-3 3 carbon atoms; alkenyl and alkynyl groups including groups having one or more unsaturated linkages and from 2 to about 12 carbon atoms or from 2 to about 6 carbon atoms; alkoxy groups having those having one or more oxygen linkages and from 1 to about 12 carbon atoms or 1 to about 6 carbon atoms; aryloxy such as phenoxy; alkylthio groups including those moieties having one or more thioether linkages and from 1 to about 12 carbon atoms or from 1 to about 6 carbon atoms; alkylsulfinyl groups including those moieties having one or more sulfinyl linkages and from 1 to about 12 carbon atoms or from 1 to about 6 carbon atoms; alkylsulfonyl groups including those moieties having one or more sulfonyl linkages and from 1 to about 12 carbon atoms or from 1 to about 6 carbon atoms; aminoalkyl groups such as groups having one or more N atoms and from 1 to about 12 carbon atoms or from 1 to about 6 carbon atoms; carbocyclic carbocyclic aryl having 6 or more carbons, particularly phenyl (e.g., R being a substituted or unsubstituted biphenyl moiety); and aralkyl such as benzyl.
The compounds of the present invention can be prepared synthetically from the intermediate compound 11 described in the '362 patent. Numerous active antitumor compounds have been prepared from this compound and it is believed that many more compounds may be formed in accordance with the teachings of the present disclosure.
One especially preferred embodiment of the present invention is the novel ecteinascidin-like compounds that have been prepared from Compound 1:
Figure USRE041614-20100831-C00006
    • wherein X1 and X2 are each independently selected from the group consisting of:
      Figure USRE041614-20100831-C00007
      or the formula:
      Figure USRE041614-20100831-C00008
    • wherein Z is selected from the group consisting of:
      Figure USRE041614-20100831-C00009
    • where n=1, 2, 3, . . . 20
      Figure USRE041614-20100831-C00010
    • wherein each R group, which may be the same or be different, is selected from the group consisting of H, OH, SH, NH2, NO2, CN, NH(C═O)CH3, O(C═O) CH3, halogen, C1-C6-alkyl, C1-C6-alkoxy, C1-C6-acyl, aryl or alkylaryl.
Another especially preferred embodiment of the present invention is pharmaceutical compositions useful as antitumor agents, comprising an effective antitumor amount of one or more of the compounds of the present invention and a pharmaceutically acceptable diluent, carrier or excipient.
Yet another especially preferred embodiment of the present invention is the synthetic intermediates of the compounds of the present invention as described in detail below.
Finally, the present invention includes the synthetic processes described herein.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
The currently most preferred compound of the present invention is the compound of formula 7:
Figure USRE041614-20100831-C00011
The preferred method of producing the compound of formula 7 is set forth below in Scheme I:
Figure USRE041614-20100831-C00012
Figure USRE041614-20100831-C00013
Figure USRE041614-20100831-C00014
Figure USRE041614-20100831-C00015
As illustrated in Scheme I, the first step for producing the preferred compound 7 of the present invention is the high yield conversion (93%) of the phenol compound 1 to the allyl ether compound 2. The second step is the high yield (99%) removal of the TBDPS protecting group to form the free alcohol compound 3. The third step in this process is the high yield (91%) coupling of phthalimide to the free alcohol compound 3 to yield the phthalimide derivative, compound 4. The phthalimide compound 4 is then converted in high yield (97%) to the phenol compound 5. Phenol compound 5 is converted in high yield (94%) to the methoxymethyl ether compound 6. Alternatively, the phthalimide compound 4 can be treated with several reagents to produce in high yield (91%) the methoxymethyl ether compound 6. The methoxymethyl ether compound 6 is finally reacted with trifluoroacetic acid to provide the desired compound 7, in high yield (94%). The overall yield of this process is about 72%.
The Scheme I method can be modified for the preparation of a preferred group of compounds. This modification is shown below in Scheme II:
Figure USRE041614-20100831-C00016
Figure USRE041614-20100831-C00017
In Scheme II, the free alcohol compound 3 is protected by reaction with 2-methoxypropene to yield the allyl ether compound 8 in high yield (99%). Compound 8 is then converted into the intermediate alcohol 9 in three steps with an overall yield of 89%. Compound 9 can be reacted with a wide variety of phthalimides, dicarboximides, or equivalents thereof (e.g., amides, including aromatic amides, ureas, urethanes, sulfonamides, alkoxy compounds, urethanes, and the like) to form compounds of the formula:
Figure USRE041614-20100831-C00018
wherein X1 is the radical provided by the phthalimide, dicarboxyimide or equivalent compound. Especially preferred compounds prepared by the Scheme II process include the compounds wherein X1 has the formula:
Figure USRE041614-20100831-C00019
and wherein Z is selected from the group consisting of:
Figure USRE041614-20100831-C00020
    • wherein each R group, which may be the same or be different, is selected from the group consisting of hydrogen, amino, halogen, nitro, C1-C6-alkyl, C1-C6-alkoxy, C1-C6-acyl, aryl, especially phenyl or alkylaryl, specially benzyl.
In yet another preferred modification, the phenol compound 5 may be transformed into a number of derivatives, as shown in Scheme III:
Figure USRE041614-20100831-C00021
As shown in Scheme III, the phenol compound (5) is reacted with various side-chain modifying carboxylic acids to afford the corresponding phenolic esters. Scheme III can be used to produce numerous compounds having the formula:
Figure USRE041614-20100831-C00022
    • wherein X2 is the radical provided by the carboxylic acid. Especially preferred X2 groups are selected from the group consisting of:
      Figure USRE041614-20100831-C00023
Another modification is the alkylation reaction illustrated in Scheme IV:
Figure USRE041614-20100831-C00024
In Scheme IV, the phenol compound 5 is treated with an alkylating agent to afford the corresponding R4 derivatives. Scheme IV can be used to produce numerous compounds having the formula:
Figure USRE041614-20100831-C00025
    • wherein X2 is the radical provided by the alkylating agent. Representative derivatives of this type include the compounds wherein X2 is selected from the group consisting of:
      Figure USRE041614-20100831-C00026
Several key intermediate compounds include the tosylate 29, the azide compound 30, and the free amine compound 31. The reaction sequence for these compounds is shown below in Scheme V:
Figure USRE041614-20100831-C00027
Figure USRE041614-20100831-C00028
The following additional compounds of the present invention (including for example, Compounds 43, 44, 45, 46, 47, 48, 49, 50, 51, 54, 55 and 56) have been prepared as described in detail in the Examples infra:
Figure USRE041614-20100831-C00029
Figure USRE041614-20100831-C00030
Figure USRE041614-20100831-C00031
Figure USRE041614-20100831-C00032
Figure USRE041614-20100831-C00033
As the skilled artisan will readily appreciate, the reaction schemes described herein may be modified and/or combined in various ways, and the compounds generated therefrom are to be considered as being part of this invention.
The present invention will be further illustrated with reference to the following examples which aid in the understanding, but which are not to be construed as limitations thereof. All percentages reported herein, unless otherwise specified, are percent by weight. All temperatures are expressed in degrees Celsius.
EXAMPLE 1
Figure USRE041614-20100831-C00034
Cesium carbonate (100.0 mg, 0.307 mmol) was gently flame dried and added as a solid to a solution of the phenol (1) (79.0 mg, 0.104 mmol) in DMF (5.5 mL). Allyl bromide (35.0 μL, 0.405 mmol) was then charged into the solution and the reaction was stirred at 23° C. for 2 h. The reaction was diluted with 1:1 ethyl acetate-hexane (100 mL), washed with water (3×100 mL), dried over sodium sulfate, decanted and concentrated in vacuo to afford Compound 2 as a pure clear viscous oil (77.2 mg, 93%). If necessary the material can be purified by flash column chromatography (70 mL silica gel, 1:2 ethyl acetate-hexane). m.p.: 167° (dec.); Rf0.57 (1:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.62-7.58 (m, 2H), 7.46-7.34 (m, 6H), 7.32-7.26 (m, 2H), 6.70 (s, 1H), 6.12 (m, 1H), 5.78 (d, J=1.5 Hz, 1H), 5.64 (d, J=1.5 Hz, 1H), 5.41 (dq, J=17.2, 1.4 Hz, 1H), 5.27 (dd, J=10.4, 1.5 Hz, 1H), 5.13 (dd, J=7.2, 5.9 Hz, 2H), 4.46 (d, J=2.6 Hz, 1H), 4.25 (d, J=1.9 Hz, 1H), 4.21-4.04 (m, 3H), 3.75 (s, 3H), 3.64 (dd, J=9.9, 2.3 Hz, 1H), 3.60 (s, 3H), 3.42-3.36 (m, 2H), 3.30-3.22 (m, 2H), 3.04 (dd, J=17.8, 8.2 Hz, 1H), 2.72 (d, J=17.8 Hz, 1H), 2.33 (s, 3H), 2.26 (s, 3H). 2.11 (s, 3H), 1.94 (dd, J=16.0, 12.2 Hz, 1H), 0.87 (s, 9H); 13C NMR (100 MHz, CDCl3) δ148.5, 148.3, 148.2, 144.1, 139.1, 135.7, 135.4, 133.8, 133.1, 132.7, 130.5, 130.4, 129.6, 129.5, 127.6, 127.5, 125.2, 124.3, 121.6, 118.5, 117.5, 113.0, 111.8, 100.9, 99.2, 74.1, 67.7, 61.5, 59.7, 59.0, 57.1, 57.2, 55.4, 41.6, 26.6, 26.5, 25.6, 18.9, 15.8, 9.2; FTIR (neat) 2931 (s br), 2857 (m), 1460 (m), 1447 (m, br), 1429 (s), 1158 (m), 1107 (s), 1093 (s), 1022 (m), 999 (m br), 931 (m, br) cm−1; HRMS (FAB), [m+Na]/z calc'd for C47H55O7N3SiNa; 824.3707, found 824.3708; [α]D 23=+73.1° (c 1.0, methylene chloride).
EXAMPLE 2
Figure USRE041614-20100831-C00035
Compound 2 (77.2 mg, 0.096 mmol) was dissolved in THF (8.0 mL) and a 1.0 M tetrabutylammonium fluoride solution in THF (200 μL, 0.20 mmol) was added. After stirring at 23° C. for 7 h the reaction was concentrated in vacuo at 23° C. The reaction was diluted into ethyl acetate/hexane (1:1, 100 mL), washed with water (3×100 mL), dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by flash column chromatography (30 mL silica gel, gradient 1:3 to 1:1 ethyl acetate-hexane) to afford Compound 3 as a clear film (53.3 mg, 99%). Rf0.28 (1:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ6.71 (s, 1H), 6.16-6.06 (m, 1H), 5.92 (d, J=1.4 Hz, 1H), 5.87 (d, J=1.4 Hz, 1H), 5.42 (dq, J=17.1, 1.4 Hz, 1H), 5.28 (dd, J=10.3, 1.3 Hz, 1H), 5.12 (s, 2H), 4.26 (d, J=2.3 Hz, (1H), 4.19 (dd, J=12.1, 5.6 Hz, 1H), 4.14 (dd, J=12.1, 6.3 Hz, 1H), 4.05 (d, J=2.5 Hz, 1H), 3.97 (t, J=3.1 Hz, 1H), 3.70 (s, 3H), 3.65 (dt, J=11.4, 2.4 Hz, 1H), 3.58 (s, 3H), 3.46 (dt, J=10.6, 2.6 Hz, 1H), 3.39-3.33 (m, 2H), 3.24 (dd, J=15.8, 2.7 Hz, 1H), 3.12 (dd, J=17.9, 7.9 Hz, 1H), 2.51 (d, J=18.1 Hz, 1H), 2.36 (s, 3H), 2.21 (s, 3H), 2.11 (s, 3H), 1.87-1.68 (m, 2H); 13C NMR (100 MHz, CDCl3) δ148.7, 148.6, 148.5, 144.4, 139.0, 133.8, 131.1, 129.5, 125.1, 124.0, 120.8, 117.6, 117.4, 113.3, 112.3, 101.1, 99.2, 74.1, 63.4, 60.0, 59.7, 58.0, 57.7, 57.1, 56.6, 55.3, 41.6, 26.2, 25.7, 15.7, 9.2; FTIR (neat) 3495 (w br), 2934 (m br), 2253 (w), 1448 (m), 1432 (m br), 1340 (m), 1158 (m), 1104 (s br), 1065 (m), 998 (m), 917 (m br) cm−1; HRMS (FAB), [m+Na]/z calc'd for C31H37O7N3Na: 586.2529, found 586.2543; [α]D 23=+96.1° (c 1.0, methylene chloride).
EXAMPLE 3
Figure USRE041614-20100831-C00036
Alcohol (3) (61.5 mg, 0.109 mmol) and phthalimide (18.8 mg, 0.128 mmol) were azeotropically dried with toluene (2×5 mL) and dissolved in THF (3.8 mL). Triphenylphosphine (35.0 mg, 0.133 mmol) was added followed by diethyl azodicarboxylate (19.0 μL, 0.121 mmol). The reaction turned yellow and then a bright orange color within 5 minutes. After stirring at 23° C. for 2 h the reaction was concentrated in vacuo at 23° C. The residue was purified by flash column chromatography (60 mL silica gel, gradient 2:1 diethyl ether-hexane to 2:3 to 1:1 ethyl acetate-hexane) to afford Compound 4 as a white foam (68.5 mg, 91%). Rf0.56 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.71-7.65 (m, 4H), 6.63 (s, 1H), 6.08 (m, 1H), 5.61 (d, J=1.5 Hz, 1H), 5.38 (dd, J=17.2, 1.6 Hz, 1H), 5.25-5.23 (m, 2H), 5.07 (dd, J=7.6, 6.0 Hz, 2H), 4.24-4.20 (m, 2H), 4.15-4.13 (m, 3H), 3.61 (d, J=5.6 Hz, 2H), 3.57 (s, 3H), 3.55 (s, 3H), 3.37 (dd, J=8.2, 5.5 Hz, 1H), 3.23 (dd, J=15.4, 2.2 Hz, 1H), 3.18 (dt, J=11.6, 2.6 Hz, 1H), 3.05 (dd, J=18.1, 8.1 Hz, 1H), 2.69 (d, J=18.1 Hz, 1H), 2.31 (s, 3H), 2.19 (s, 3H), 2.10 (s, 3H), 1.69 (dd, J=15.3, 11.6 Hz, 1H); 13C NMR (100 MHz, CDCl3) δ167.7, 151.3, 148.7, 148.3, 148.1, 144.2, 139.5, 133.8, 133.5, 131.9, 130.3, 130.2, 125.1, 123.8, 122.9, 121.0, 118.0, 117.5, 113.6, 112.4, 100.8, 99.2, 74.3, 60.3, 59.6, 57.7, 57.5, 56.9, 55.7, 55.5, 41.9, 41.5, 26.6, 25.4, 16.0, 9.4; FTIR (neat) 2935 (m, br), 2256 (w), 1773 (m), 1716 (s), 1459 (m br), 1432 (m br), 1343 (m), 1267 (m br), 1233 (m), 1158 (m), 1100 (s), 1064 (m), 1024 (m), 947 (m br) cm−1; HRMS (FAB), [m+Na]/z calc'd for C39H40O8N4Na: 715.2744, found 715.2730; [α]D 23=+72.7° (c 1.0, methylene chloride).
EXAMPLE 4
Figure USRE041614-20100831-C00037
Phthalimide (4) (20.0 mg, 0.0289 mmol) and acetic acid (16.5 μL, 0.289 mmol) were dissolved in methylene chloride (0.8 mL). PdCl2(PPh3)2 (1.0 mg, 1.4 μmol) was added followed by tributyltin hydride (21.0 μL, 0.0779 mmol). Bubbling was observed and the reaction changes from a yellow to a dark orange color. After stirring at 23° C. for 10 min the reaction was quenched into water (20 mL), extracted with methylene chloride (2×20 mL), dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by flash column chromatography (30 mL silica gel, gradient 1:4 to 1:1 to 2:1 ethyl acetate-hexane) to afford Compound 5 as a white foam (18.3 mg, 97%). Rf0.42 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.75-7.71 (m, 2H), 7.69-7.65 (m, 2H), 6.61 (s, 1H), 5.51 (s, 1H), 5.27 (d, J=6.0 Hz, 1H), 5.23 (br s, 1H), 5.13 (d, J=6.0 Hz, 1H), 5.06 (s, 1H), 4.25 (d, J=2.4 Hz, 1H), 4.21 (d, J=5.0 Hz, 1H), 4.16 (d, J=2.1 Hz, 1H), 3.67 (s, 3H), 3.66 (s, 3H), 3.53 (m, 2H), 3.37 (d, J=7.8 Hz, 1H), 3.22 (d, J=11.5 Hz, 1H), 3.15 (d, J=14.7 Hz, 1H), 3.05 (dd, J=18.0, 8.1 Hz, 1H), 2.65 (d, J=18.0 Hz, 1H), 2.31 (s, 3H), 2.20 (s, 3H), 2.05 (s, 3H), 1.73 (m, 1H); 13C NMR (100 MHz, CDCl3) δ167.9, 148.7, 147.5, 145.6, 145.5, 144.3, 136.9, 133.6, 132.0, 130.5, 124.9, 123.0, 117.9, 113.1, 112.4, 106.3, 100.5, 99.6, 60.3, 59.8, 57.7, 57.0, 56.7, 55.5, 55.3, 42.4, 41.6, 25.9, 25.4, 15.9, 8.9; FTIR (neat) 3464 (w br), 2936 (w br), 1773 (w), 1715 (s), 1484 (w), 1461 (m), 1433 (m), 1397 (m), 1235 (w), 1157 (w), 1101 (m), 1076 (w), 1060 (w), 1023 (w), 1007 (w), 957 (w) cm−1; HRMS (FAB), [m+H]/z calc'd for C36H37O8N4: 653.2611, found 653.2608; [α]D 23=+3.1° (c 0.35, methylene chloride).
EXAMPLE 5
Figure USRE041614-20100831-C00038
Phenol (5) (1.1 mg, 0.0017 mmol) was dissolved in methylene chloride (0.15 mL). 4-Dimethylaminopyridine (0.5 mg, 0.0041 mmol) and acetic anhydride (0.5 μL, 0.0053 mmol) were added to the solution which was stirred at 23° C. for 30 min. The reaction mixture was concentrated in vacuo. The residue was purified by flash column chromatography (0.5 mL silica gel, gradient 1:4 to 1:1 to 2:1 ethyl acetate-hexane) to afford Compound 6 (1.1 mg, 94%). Rf0.53 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.70-7.63 (m, 4H), 6.64 (s, 1H), 5.73 (s, 1H), 5.50 (s, 1H), 5.07 (d, J=5.7 Hz, 1H) 4.98 (d, J=5.7 Hz, 1H), 4.27 (d, J=2.1 Hz, 1H), 4.24 (m, 1H), 4.08 (d, J=2.5 Hz, 1H), 3.74-3.67 (m, 2H), 3.53 (s, 3H), 3.50 (s, 3H), 3.38 (d, J=7.1 Hz, 1H), 3.18 (d, J=11.5 Hz, 1H), 3.02 (dd, J=18.1, 8.1 Hz, 1H), 2.75 (d, J=16.1 Hz, 2H), 2.31 (s, 3H), 2.27 (s, 3H), 2.18 (s, 3H), 2.01 (s, 3H), 1.60 (m, 1H); 13C NMR (100 MHz, CDCl3) δ168.3, 167.5, 148.1, 147.8, 144.3, 141.2, 140.5, 133.4, 131.8, 130.2, 125.3, 123.4, 123.0, 120.8, 118.0, 113.6, 111.7, 101.3, 99.1, 59.8, 59.6, 57.7, 56.7, 56.6, 56.1, 55.4, 41.5, 40.9, 26.7, 25.0, 20.1, 16.0, 9.5; FTIR (neat) 2935 (m br), 17641 (m), 1716 (s), 1433 (m br), 1394 (m br), 1369 (m br), 1234 (m), 1198 (s), 1158 (m), 1101 (m br), 1072 (m), 1025 (m), 1000 (m), 947 (m), 933 (m) cm−1; HRMS (FAB), [m+H]/z calc'd for C38H39O9N4: 695.2717, found 695.2744; [α]D 23+21.6° (c 1.0, methylene chloride).
EXAMPLE 6
Figure USRE041614-20100831-C00039
Alternatively, phthalimide (4) (68.5 mg, 0.99 mmol) and acetic acid (17.0 μL, 0.30 mmol) were dissolved in methylene chloride (6.0 mL). PdCl2(PPh3)2 (3.5 mg, 5 μmol) was added followed by tributyltin hydride (67.0 μL, 0.25 mmol). Bubbling was observed and the reaction changes from a yellow to a dark orange color. After stirring at 23° C. for 10 min to the reaction was added triethylamine (55.0 μL, 0.40 mmol), 4-dimethylaminopyridine (5.5 mg, 0.045 mmol) and acetic anhydride (38.0 μL, 0.39 mmol). The reaction was stirred at 23° C. for 10 min and was quenched into quarter-saturated aqueous sodium chloride solution (20 mL), extracted with methylene chloride (3×20 mL), dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by flash column chromatography (40 mL silica gel, 1:1 ethyl acetate-hexane) to afford Compound 6 as a white foam (62.8 mg, 91%).
EXAMPLE 7
Figure USRE041614-20100831-C00040
The methoxymethyl ether (6) (3.8 mg, 0.00547 mmol) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 4.0 mL) and the solution was stirred at 23° C. for 7 h. The reaction mixture was diluted with toluene (5 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×5 mL). The residue was dissolved in ethyl acetate (10 mL) and washed with a saturated aqueous sodium bicarbonate solution (20 mL), the aqueous layer was extracted with ethyl acetate (2×10 mL) and the combined organic layers were dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by flash column chromatography (1.5 mL silica gel, gradient methylene chloride to 2:1 ethyl acetate-hexane) to afford Compound 7 (3.4 mg, 94%). Rf0.41 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ7.73-7.71 (m, 2H), 7.67-7.65 (m, 2H), 6.39 (s, 1H), 5.66 (s, 1H), 5.59 (s, 1H), 5.33 (br s, 1H), 4.25-4.23 (m, 2H), 4.02 (d, J=2.5 Hz, 1H), 3.64 (m, 5H), 3.35 (d, J=8.3 Hz, 1H), 3.20 (d, J=12.0 Hz, 1H), 3.02 (dd, J=18.1, 8.1 Hz, 1H), 2.77 (d, J=14.6 Hz, 1H), 2.45 (d, J=18.1 Hz, 1H), 2.29 (s, 6H), 2.22 (s, 3H), 1.99 (s, 3H), 1.73 (t, J=14.3 Hz, 1H); 13C NMR (125 MHz, CDCl3) δ167.7, 146.3, 144.3, 142.6, 141.2, 140.6, 133.5, 131.9, 130.9, 128.3, 123.1, 121.0, 120.9, 118.0, 116.5, 113.7, 111.8, 101.2, 60.5, 60.2, 57.1, 56.4, 55.6, 55.5, 41.8, 41.6, 26.6, 25.3, 20.3 15.9, 9.6; FTIR (neat) 3463 (m br), 2934 (m br), 1764 (m), 1716 (s), 1455 (m br), 1433 (m br), 1395 (m br), 1370 (m), 1233 (m), 1102 (m), 1073 (m) cm−1; HPLC (Columbus, 5μ, C18, 100 Å, 250×4.60 mm, flow rate: 1.0 mL/min, λ=254 nm), RT=13.7 min (60% CH3CN in water); HRMS (FAB), [m+H]/z calc'd for C36H35O8N4: 651.2455, found 651.2444; [α]D 23=+21.9° (c 1.0, methylene chloride).
EXAMPLE 8
Figure USRE041614-20100831-C00041
Alcohol (4) (31.3 mg, 0.056 mmol) was dissolved in 2-methoxypropene. Catalytic phosphorus oxychloride was added and stirred at 23° C. for 15 min. One drop of triethylamine and methanol (1 mL) were added to quench the reaction which was then concentrated vacuo. The residue was purified by flash column chromatography (2 mL silica gel, gradient methylene chloride to 1:1 ethyl acetate-hexane) to afford Compound 8 (35.0 mg, 99%). Rf0.48 (1:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ6.62 (s, 1H), 6.17-6.06 (m, 1H), 5.92 (s, 1H), 5.85 (s, 1H), 5.41 (d, J=17.2 Hz, 1H), 5.27 (d, J=10.4 Hz, 1H), 5.13-5.08 (m, 2H), 4.41 (s, 1H), 4.23-4.10 (m, 3H), 4.04 (d, J=8.2 Hz, 1H), 3.73 (s, 3H), 3.43 (s, 3H), 3.42 (dd, J=8.8, 2.6 Hz, 1H), 3.29 (d, J=7.7 Hz, 1H), 3.22 (d, J=14.1 Hz, 2H), 3.07-2.96 (m, 4H), 2.84 (t, J=8.9 Hz, 1H), 2.64 (d, J=17.6 Hz, 1H), 2.31 (s, 3H), 2.12 (s, 3H), 2.02 (s, 3H), 1.82 (dd, J=15.3, 12.0 Hz, 1H), 1.29 (s, 3H), 1.17 (s, 3H); 13C NMR (100 MHz, CDCl3) δ148.5, 148.3, 148.1, 144.2, 139.3, 133.8, 130.8, 130.2, 124.8, 124.2, 121.4, 118.9, 117.6, 113.0, 112.0, 101.0, 99.8, 99.2, 74.2, 67.0, 62.0, 59.7, 57.7, 57.4, 57.3, 56.7, 55.5, 48.3, 41.6, 26.3, 25.6, 24.4, 24.3, 15.7, 9.3; FTIR (neat) 2988 (w), 2933 (m br), 2825 (w), 1483 (m), 1460 (m), 1444 (m), 1432 (m), 1421 (m), 1380 (m), 1367 (w), 1341 (w), 1232 (w), 1212 (m), 1157 (m), 1104 (s), 1094 (s), 1076 (m), 1066 (m), 1046 (m), 1023 (m), 999 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C35H45O8N3Na: 658.3104, found 658.3114; [α]D 23+107° (c 0.10, methylene chloride).
EXAMPLE 9
Figure USRE041614-20100831-C00042
Allyl ether (8) (35.0 mg, 0.055 mmol) and acetic acid (20.0 μL, 0.35 mmol) were dissolved in methylene chloride (2.0 mL). PdCl2(PPh3)2 (2.5 mg, 0.0036 mmol) was added as a solid followed by tributyltin hydride (40.0 μL, 0.148 mmol). Bubbling was observed and the reaction changes color from a yellow to a dark orange. After stirring at 23° C. for 5 min, triethylamine (100 μL, 0.72 mmol), 4-dimethylaminopyridine (7.0 mg, 0.057 mmol) and acetic anhydride (10.0 μL, 0.10 mmol) were added to the solution. After stirring at 23° C. for 10 min, the reaction was concentrated in vacuo and dissolved in a solution of 19:1 acetic acid-water (2.0 mL). After stirring at 23° C. for 5 min the reaction was concentrated in vacuo and the residue was purified by flash column chromatography (14 mL silica gel, gradient 1:1 to 2:1 ethyl acetate-hexane) to afford Compound 9 (27.8 mg, 89%). Rf0.19 (1:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ6.70 (s, 1H), 5.96 (d, J=1.3 Hz, 1H), 5.90 (d, J=1.4 Hz, 1H), 5.14 (d, J=5.7 Hz, 1H), 5.07 (d, J=5.7 Hz, 1H), 4.21 (d, J=2.3 Hz, 1H) 4.10 (d, J=1.8 Hz, 1H), 3.99 (t, J=3.3 Hz, 1H), 3.72 (s, 3H), 3.66 (d, J=11.1 Hz, 1H), 3.58 (s, 3H), 3.49-3.44 (m, 1H), 3.40-3.32 (m, 2H), 3.10 (dd, J=18.0, 7.9 Hz, 1H), 2.79 (d, J=15.7 Hz, 1H), 2.51 (d, J=18.1 Hz, 1H), 2.36 (s, 3H), 2.32 (s, 3H), 2.21 (s, 3H), 2.00 (s, 3H), 1.82-1.70 (m, 2H); 13C NMR (125 MHz, CDCl3) δ168.5, 148.6, 148.3, 144.5, 140.6, 140.4, 131.3, 129.5, 125.1, 123.6, 120.5, 117.6, 113.2, 111.7, 101.5, 99.2, 63.6, 59.9, 59.8, 58.0, 57.7, 56.9, 56.1, 55.3, 41.6, 26.3, 25.6, 20.1, 15.7, 9.3; FTIR (neat) 3500 (m br), 2935 (s br), 2854 (w), 1760 (s), 1484 (m), 1440 (m), 1434 (m), 1401 (m), 1370 (m), 1341 (w), 1324 (w), 1234 (m), 1201 (s), 1158 (m), 1106 (s), 1086 (s), 1075 (s), 1043 (m), 1023 (m), 1000 (m), 961 (m), 912 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C30H35O8N3Na: 588.2322, found 588.2303; [α]D 23+50.2° (c 0.66, methylene chloride).
Methods for Generating the Dicarboximides
Many of the phthalimides and dicarboximides were not commercially available and had to be synthesized from the commercially available anhydrides or dicarboxylic acids using a variety of established methodologies. The dicarboxylic acids were converted to the anhydrides by heating with acetic anhydride. Heating the anhydrides with urea1, urethane2 or formamide3 at
1 Campayo, L.; Jimenez, B.; Manzano, T.: Navarro, P. Synthesis 1985, 197 and Crockett, G. C.; Swanson, B. J.; Anderson, D. R.; Kock, T. H. Synth. Commun. 1981, 11 (6), 447-454.
2 Weider-Wells, M. A.; DeCamp, A.; Mazzocchi, P. H. J. Org. Chem. 1989, 54 (24), 5746-5758.
3 Vostrova, V. N.; Plakidin, V. L. J. Org. Chem. USSR 1982, 18, 1754 and Ganin, E. V.; Makarov, V. F.; Nikitin, V. I. J. Org. Chem. USSR 1987, 23, 981-983. ˜200° C. (15 minutes to 12 hours) and crystallization from water afforded pure to semi-pure dicarboximides. Filtration through a pad of silica gel and elution with ethyl acetate provided pure material. Alternatively the anhydrides were reacted with ammonium hydroxide followed by refluxing in ethanol with catalytic hydrochloric acid4. The 1,2-Naphthalimide was synthesized via a Diels-Alder with β-bromostyrene and maleimide.5 p-Toluenesulfonyl isocyanate and t-butanol were reacted in order to generate the BOC-protected tolylsulfonamide.6 The dicarboximides were systematically dried under vacuum (60° C., 30 mm) and by toluene azeotrope immediately before use.
4 Alexion, M.; Tyman, J.; Wilson, I. Tetrahedron Lett. 1981, 22 (24), 2303.
5 Newman, M. S.; Dhawan, B.; Hehem, M. M.; Khanna, V. K.; Springer, J. M. J. Org. Chem. 1976, 41 (24), 3925.
6 Corey, E. J.; Su, Wei-juo Tetrahedron Lett. 1990, 31 (27), 3833-3836.
EXAMPLE 10
General Procedure for the Mitsunobu Coupling Reaction of Alcohol (9) with Dicarboximides
Figure USRE041614-20100831-C00043
The alcohol (9) (1.0 mg, 0.0018 mmol) and the dicarboximide (0.0065 mmol, 3.6 equiv.) were azeotropically dried with toluene (2×0.1 mL) and dissolved in THF (0.2 mL). Triphenylphosphine (1.7 mg, 0.0065 mmol) was added as a solid followed by diethyl azodicarboxylate (1.0 μL, 0.0064 mmol) via syringe. The reaction turned yellow and after stirring at 23° C.7 for 15 h the reaction was concentrated in vacuo. The residue was purified by flash column chromatography (1.0 mL silica gel, gradient methylene chloride to 2:1 diethyl ether-hexane to 1:1 to 2:1 ethyl acetate-hexane) followed by preparative thin layer chromatography to afford the desired product.
The methoxymethyl ether was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.2 mL) and the solution was stirred at 23° C. for 11 h. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by flash column chromatography (0.5 mL silica gel, gradient methylene chloride to 2:1-ethyl acetate-hexane) to afford the desired product.
TABLE 1
General Procedure for the Coupling of Alcohol (9) with
Dicarboximides.
Com- Mitsunobu MOM
pound Coupling Removal
Entry # Dicarboximide Yield (%) Yield (%)
1 10
Figure USRE041614-20100831-C00044
 		56 100
2 11
Figure USRE041614-20100831-C00045
 		51 40
3 12
Figure USRE041614-20100831-C00046
 		89 53
4 13
Figure USRE041614-20100831-C00047
 		53 91
5 14
Figure USRE041614-20100831-C00048
 		61 93
6 15
Figure USRE041614-20100831-C00049
 		76 96
7 16
Figure USRE041614-20100831-C00050
 		82 25
8 17
Figure USRE041614-20100831-C00051
 		92 90
9 18
Figure USRE041614-20100831-C00052
 		100 98
10 19
Figure USRE041614-20100831-C00053
 		34 100
7 Heating to 40° C. was required for entries 8 and 9.
EXAMPLE 11
Compound 10—Preparative thin layer chromatography of the first step done using 4:1 diethyl ether-hexane. Rf0.42 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ6.50 (s, 1H), 5.99 (s, 1H), 5.91 (s, 1H), 5.62 (s, 1H), 4.28 (s, 1H), 4.16 (d, J=3.1 Hz, 1H), 4.02 (s, 1H), 3.76 (s, 3H), 3.75-3.70 (m, 2H), 3.37 (d, J=7.3 Hz, 1H), 3.15 (d, J=11.4 Hz, 1H), 2.96 (dd, J=18.0, 7.9 Hz, 1H), 2.86 (d, J=18.0 Hz, 1H), 2.74 (d, J=15.5 Hz, 1H), 2.43 (q, J=7.4 Hz, 1H), 2.29 (s, 6H), 2.26 (s, 3H), 2.01 (s, 3H), 2.04-2.02 (m, 1H), 1.80-1.45 (m, 4H), 1.40-1.17 (m, 5H); FTIR (neat) 3412 (m br), 2935 (m br), 2858 (m), 2256 (w), 1759 (m), 1706 (s), 1498 (w), 1452 (m), 1434 (m), 1396 (m), 1370 (m), 1334 (m), 1325 (m), 1295 (m), 1234 (m), 1201 (m), 1148 (m), 1105 (m), 1093 (m), 1075 (m), 1008 (m), 913 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H40O8N4Na: 679.2744, found 679.2727.
EXAMPLE 12
Compound 11—Preparative thin layer chromatography of the first step was done using 4:1 diethyl ether-hexane. Rf0.45 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ8.85 (d, J=8.7 Hz, 1H), 8.12 (d, J=8.2 Hz, 1H), 7.95 (d, J=8.1 Hz, 1H), 7.76-7.72 (m, 2H), 7.67-7.65 (m, 1H), 6.26 (s, 1H) 5.63 (s, 1H), 5.58 (s, 1H), 5.34 (br s, 1H), 4.33-4.28 (m, 2H), 4.07 (s, 1H), 3.72-3.65 (m, 2H), 3.57 (s, 3H), 3.40 (d, J=8.0 Hz, 1H), 3.25 (d, J=11.5 Hz, 1H), 3.02 (dd, J=17.1, 7.6 Hz, 1H), 2.80 (d, J=14.9 Hz, 1H), 2.66 (d, J=18.6 Hz, 1H), 2.31 (s, 3H), 2.30 (s, 3H), 2.00 (s, 3H), 1.99 (s, 3H), 1.80 (dd, J=14.9, 11.7 Hz, 1H); FTIR (neat) 3438 (m br), 2938 (m br), 1763 (m), 1706 (s), 1588 (w), 1500 (w), 1456 (m), 1431 (m), 1388 (m), 1231 (m), 1200 (m), 1144 (w), 1100 (m), 1075 (m), 1031 (w), 1006 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C40H36O8N4Na: 723.2431, found 723.2443.
EXAMPLE 13
Compound 12—Preparative thin layer chromatography of the first step was done using 1:1 ethyl acetate-hexane. Rf0.34 (1:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ7.73 (s, 2H), 6.40 (s, 1H), 5.78 (s, 1H), 5.63 (br s, 1H), 5.55 (s, 1H), 4.24-4.21 (m, 2H), 4.00 (d, J=1.9 Hz, 1H), 3.74-3.71 (m, 2H), 3.60 (s, 3H), 3.36 (d, J=8.1 Hz, 1H), 3.18 (d, J=11.9 Hz, 1H), 3.00 (dd, J=17.9, 8.2 Hz, 1H), 2.75-2.69 (m, 2H), 2.28 (s, 6H), 2.24 (s, 3H), 2.01 (s, 3H), 1.61-1.54 (m, 1H); FTIR (neat) 3415 (m br), 2933 (m br), 2855 (w), 1762 (m), 1720 (s), 1500 (w), 1459 (m), 1452 (m), 1433 (m), 1387 (m), 1369 (m), 1265 (m), 1234 (m), 1196 (m), 1144 (m), 1102 (m), 1083 (m), 1074 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H32O8N4Cl2Na: 741.1495, found 741.1498.
EXAMPLE 14
Compound 13—Preparative thin layer chromatography of the first step was done using 4:1 diethyl ether-hexane and again using 1:1 ethyl acetate-hexane. Rf0.20 (1:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ8.21 (s, 2H), 8.03 (dd, J=6.1, 3.2 Hz, 2H), 7.69 (dd, J=6.1, 3.2 Hz, 2H), 6.38 (s, 1H), 5.62 (s, 1H), 5.57 (s, 1H), 5.30 (s, 1H), 4.31-4.28 (m, 2H), 4.02 (s, 1H), 3.73-3.68 (m, 2H), 3.52 (s, 3H), 3.36 (d, J=7.3 Hz, 1H), 3.22 (d, J=11.7 Hz, 1H), 3.02 (dd, J=18.2, 7.7 Hz, 1H), 2.78 (d, J=15.3 Hz, 1H), 2.67 (d, J=18.0 Hz, 1H), 2.30 (s, 3H), 2.29 (s, 3H), 2.13 (s, 3H), 1.99 (s, 3H), 1.78 (dd, J=14.8, 12.4 Hz, 1H); FTIR (neat) 3428 (m br), 2983 (m br), 1766 (m), 1712 (s), 1432 (m), 1384 (m), 1197 (m), 1150 (w), 1103 (m), 905 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C40H36O8N4Na: 723.2431, found 723.2416.
EXAMPLE 15
Compound 14—Preparative thin layer chromatography of the first step was done using 4:1 diethyl ether-hexane. Rf0.20 (4:1 diethyl ether-hexane); 1H NMR (400 MHz, CDCl3) δ8.50 (d, J=7.9 Hz, 1H), 8.46 (s, 1H), 7.85 (d, J=8.1 Hz, 1H), 6.43 (s, 1H), 5.76 (s, 1H), 5.58 (br s, 1H), 5.54 (s, 1H), 4.27 (t, J=4.6 Hz, 1H), 4.24 (d, J=2.0 Hz, 1H), 4.00 (d, J=2.5 Hz, 1H), 3.79 (d, J=4.0 Hz, 2H), 3.57 (br s, 3H), 3.38 (d, J=8.0 Hz, 1H), 3.18 (d, J=11.6 Hz, 1H), 3.02 (dd, J=18.1, 18.1 Hz, 1H), 2.74 (d, J=16.7 Hz, 2H), 2.28 (s, 3H), 2.26 (s, 3H), 2.21 (s, 3H), 2.01 (s, 3H), 1.65-1.55 (m, 1H); FTIR (neat) 3488 (w br), 2932 (m br), 1761 (m), 1725 (s), 1622 (w), 1584 (w), 1541 (m), 1499 (w), 1435 (m), 1393 (w), 1345 (m), 1233 (m), 1196 (m), 1146 (w), 1105 (m), 1075 (m), 1030 (m), 1001 (w), 951 (w), 907 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H33O10N5Na: 718.2125, found 718.2125.
EXAMPLE 16
Compound 15—Preparative thin layer chromatography of the first step was done using 1:1 ethyl acetate-hexane. Rf0.25 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ8.07 (dd, J=8.1, 0.7 Hz, 1H), 7.96 (dd, J=7.5, 0.8 Hz, 1H), 7.84 (t, J=7.8 Hz, 1H), 6.36 (s, 1H), 5.68 (s, 1H), 5.60 (s, 1H), 5.46 (br s, 1H), 4.30-4.20 (m, 2H), 4.03 (d, J=1.8 Hz, 1H), 3.75-3.65 (m, 5H), 3.35 (d, J=8.4 Hz, 1H), 3.21 (d, J=12.3 Hz, 1H), 3.02 (dd, J=18.2, 8.2 Hz, 1H), 2.78 (d, J=16.5 Hz, 1H), 2.61 (d, J=17.8 Hz, 1H), 2.30 (s, 3H), 2.28 (s, 3H), 2.21 (s, 3H), 2.00 (s, 3H), 1.80-1.70 (m, 1H); FTIR (neat) 3490 (w br), 2938 (m br), 1762 (m), 1722 (s), 1543 (m), 1459 (m), 1448 (m), 1444 (m), 1433 (m), 1394 (m), 1369 (m), 1233 (m), 1196 (m), 1103 (m), 1074 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H33O10N5Na: 718.2125, found 718.2122.
EXAMPLE 17
Compound 16—Preparative thin layer chromatography of the first step was done using 2:1 ethyl acetate-hexane. Rf0.19 (2:1 ethyl acetate/hexane); 1H NMR (400 MHz, CDCl3) δ6.49 (s, 1H), 5.94 (s, 1H), 5.87 (s, 1H), 5.64 (s, 1H), 4.20 (d, J=2.2 Hz, 1H), 4.15 (t, J=4.4 Hz, 1H), 4.03 (d, J=1.2 Hz, 1H), 3.78 (s, 3H), 3.65-3.43 (m, 2H), 3.35 (d, J=7.8 Hz, 1H), 3.17 (d, J=12.3 Hz, 1H), 2.99 (dd, J=18.5, 7.9 Hz, 1H), 2.76 (dd, J=15.6, 1.8 Hz, 2H), 2.43-2.10 (m, 13H), 2.01 (s, 3H), 1.70-1.60 (m, 1H); FTIR (neat) 3428 (m br), 2926 (s br), 2853 (m), 1757 (m), 1705 (s), 1497 (w), 1431 (m br), 1233 (w), 1198 (m), 1150 (w), 1086 (m), 920 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C32H34O8N4Na: 625.2274, found 625.2274.
EXAMPLE 18
Compound 17—The Mitsunobu was conducted at 40° C. and the purification by preparative thin layer chromatography was done using 10% ethyl acetate-methylene chloride and again using 5% methanol-methylene chloride. Rf0.31 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ8.48 (m, 2H), 8.21 (dd, J=8.3, 0.9 Hz, 2H), 7.75 (t, J=7.6 Hz, 2H), 6.34 (s, 1H), 5.68 (s, 1H), 5.29 (s, 1H), 4.62 (br s, 1H), 4.46 (d, J=2.2 Hz, 1H), 4.34 (dd, J=9.4, 3.3 Hz, 1H), 4.23 (dd, J=12.7, 9.7 Hz, 1H), 4.07 (d, J=2.2 Hz, 1H), 3.90 (dd, J=13.0, 3.3 Hz, 1H), 3.79 (s, 3H), 3.35 (d, J=9.5 Hz, 1H), 3.24 (d, J=11.9 Hz, 1H), 3.04 (dd, J=18.3, 8.4 Hz, 1H), 2.85 (d, J=14.9 Hz, 1H), 2.58 (d, J=17.8 Hz, 1H), 2.34 (s, 3H), 2.30 (s, 3H), 2.24 (s, 3H), 2.02-1.95 (m, 1H), 1.94 (s, 3H); FTIR (neat) 3422 (m br), 2929 (m br), 1761 (m), 1704 (m), 1660 (s), 1591 (m), 1456 (m), 1439 (m), 1378 (m), 1236 (s), 1198 (m), 1105 (m), 1074 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C40H36O8N4Na: 723.2431, found 723.2433.
EXAMPLE 19
Compound 18—The Mitsunobu was conducted at 40° C. and the purification by preparative thin layer chromatography of the first step was done using 5% methanol-methylene chloride. Rf0.45 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ9.19 (s, 1H), 9.12 (d, J=2.2 Hz, 1H), 8.66 (d, J=6.5 Hz, 1H), 8.41 (d, J=8.3 Hz, 1H), 7.93 (t, J=7.8 Hz, 1H), 6.34 (s, 1H), 5.67 (s, 1H), 5.34 (s, 1H), 4.75 (br s, 1H), 4.42 (d, J=2.3 Hz, 1H), 4.34 (dd, J=9.3, 3.2 Hz, 1H), 4.29-4.21 (m, 1H), 4.07 (d, J=2.0 Hz, 1H), 3.95 (dd, J=13.1, 3.1 Hz, 1H), 3.77 (s, 3H), 3.37 (d, J=7.9 Hz, 1H), 3.23 (d, J=11.8 Hz, 1H), 3.06 (dd, J=18.1, 8.2 Hz, 1H), 2.84 (d, J=15.5 Hz, 1H), 2.59 (d, J=18.1 Hz, 1H), 2.33 (s, 3H), 2.30 (s, 3H), 2.22 (s, 3H), 2.03-1.85 (m, 4H); FTIR (neat) 3463 (m br), 2931 (m br), 1762 (m), 1711 (m), 1668 (s), 1600 (m), 1542 (m), 1458 (m), 1433 (m), 1420 (m), 1370 (m), 1345 (m), 1328 (m), 1234 (m), 1197 (m), 1104 (m), 1075 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C40H35O10N5Na: 768.2282, found 768.2308.
EXAMPLE 20
Compound 19—Preparative thin layer chromatography of the first step was done using 4:1 diethyl ether-hexane. Rf0.50 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ7.28 (d, J=8.2 Hz, 2H), 7.00 (d, J=8.0 Hz, 2H), 6.59 (s, 1H), 5.76 (s, 2H), 5.53 (br s, 1H), 4.11 (s, 1H), 4.00 (d, J=6.3 Hz, 1H), 3.93 (s, 1H), 3.89 (s, 1H), 3.80 (s, 3H), 3.57-3.45 (m, 1H), 3.35-3.29 (m, 2H), 3.18-3.11 (m, 2H), 2.72-2.87 (m, 1H), 2.49 (d, J=16.9 Hz, 1H), 2.39 (s, 3H), 2.36 (s, 3H), 2.33 (s, 3H), 2.32 (s, 3H), 1.99 (s, 3H), 1.82 (dd, J=16.4, 12.4 Hz, 1H); FTIR (neat) 3425 (w br), 3331 (m br), 2958 (m), 2927 (s br), 2855 (m), 1759 (s), 1719 (w), 1498 (w), 1459 (m), 1390 (m), 1370 (m), 1326 (m), 1233 (s), 1201 (s), 1154 (s), 1111 (m), 1088 (s), 1074 (s), 1028 (m), 1007 (m), 995 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C35H38O8N4SNa: 697.2308, found 697.2318.
EXAMPLE 21
Figure USRE041614-20100831-C00054
Nitro compound (14) (0.5 mg, 0.00072 mmol) was dissolved in methanol (0.4 mL), 10% Pd/C (0.2 mg) and ammonium formate (12.0 mg, 0.19 mmol) were added at 23° C. and the reaction was stirred for 40 min. The mixture was diluted with ethyl acetate (2 mL), filtered through a plug of Celite, concentrated in vacuo and the residue was purified by flash column chromatography (1.5 mL silica gel, 2:1 ethyl acetate-hexane) to afford Compound 20 (0.3 mg, 63%). Rf0.20 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) □ 7.49 (d 8.1 l), 6.91 (d 2.1, 1H), 6.77 (dd, J=8.1, 2.2 Hz, 1H), 6.38 (s, 1H), 5.67 (s, 1H), 5.61 (s, 1H), 5.34 (br s, 1H), 4.28 (br s, 2H), 4.23-4.19 (m, 2H), 4.03 (d, J=1.8 Hz, 1H), 3.71 (s, 3H), 3.53 (d, J=5.7 Hz, 2H), 3.33 (d, J=8.2 Hz, 1H), 3.20 (d, J=12.3 Hz, 1H), 3.01 (dd, J=17.6, 8.1 Hz, 1H), 2.78 (d, J=14.7 Hz, 1H), 2.61 (d, J=18.6 Hz, 1H), 2.31 (s, 3H), 2.29 (s, 3H), 2.24 (s, 3H), 1.98 (s, 3H), 1.79 (dd, J=14.4, 11.8 Hz, 1H); FTIR (neat) 3456 (w br), 3374 (m br), 3243 (w br), 2932 (m br), 2853 (w), 1760 (m), 1703 (m), 1699 (s), 1617 (m), 1501 (m), 1463 (m), 1457 (m), 1431 (m), 1398 (m), 1232 (m), 1199 (m), 1103 (m), 1073 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H35O8N5Na: 688.2383, found 688.2367.
EXAMPLE 22
Figure USRE041614-20100831-C00055
Nitro compound (18) (0.5 mg, 0.00067 mmol) was dissolved in methanol (0.4 mL), 10% Pd/C (0.2 mg) and ammonium formate (12.0 mg, 0.19 mmol) were added at 23° C. and the reaction was stirred for 40 min. The mixture was diluted with ethyl acetate (2 mL), filtered through a plug of Celite, concentrated in vacuo and the residue was purified by flash column chromatography (1.5 mL silica gel, 2:1 ethyl acetate-hexane) to afford Compound 21 (0.4 mg, 83%). Rf0.28 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ8.21 s (1), 7.93-7.91 (m, 2H), 7.59 (t, J=7.8 Hz, 1H), 7.28 (d, J=2.3 Hz, 1H), 6.35 (s, 1H), 5.68 (s, 1H), 5.32 (s, 1H), 4.67 (br s, 1H), 4.44 (s, 1H), 4.32 (dd, J=9.6, 3.2 Hz, 1H), 4.20 (t, J=11.0 Hz, 1H), 4.14 (s, 2H), 4.07 (d, J=2.3 Hz, 1H), 3.86 (dd, J=13.1, 3.3 Hz, 1H), 3.80 (s, 3H), 3.34 (d, J=8.5 Hz, 1H), 3.24 (d, J=12.1 Hz, 1H), 3.04 (dd, J=17.8, 7.9 Hz, 1H), 2.84 (d, J=14.4 Hz, 1H), 2.57 (d, J=17.6 Hz, 1H), 2.34 (s, 3H), 2.30 (s, 3H), 2.24 (s, 3H), 2.05-1.93 (m, 4H); FTIR (neat) 3456 (m br), 3369 (s br), 3250 (w br), 2931 (m br), 2856 (w), 1750 (m), 1700 (s), 1656 (s), 1619 (s), 1581 (m), 1450 (s), 1375 (m), 1331 (w), 1300 (m), 1231 (m), 1219 (m), 1150 (w), 1106 (m), 1075 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C40H37O8N5Na: 738.2540, found 738.2566.
EXAMPLE 23
Figure USRE041614-20100831-C00056
Alcohol (9) (1.0 mg, 0.0018 mmol) was dissolved in methylene chloride (0.2 mL) and 4-dimethylaminopyridine (0.1 mg, 0.00082 mmol) and phenyl isocyanate (0.5 μL, 0.0046 mmol) were added to the solution. The reaction was stirred at 23° C. for 3 hr and then quenched into a saturated solution of aqueous sodium bicarbonate (10 mL). The mixture was extracted with methylene chloride (2×5 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo to afford a residue (1.2 mg, 100%). This crude material was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL) and the solution was stirred at 23° C. for 11 hr. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by flash column chromatography (1 mL silica gel, gradient methylene chloride to 1:4 to 1:1 to 2:1 ethyl acetate-hexane) to afford Compound 22 (0.8 mg, 71%). Rf0.54 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CD2Cl2) δ7.28-7.25 (m, 4H), 7.04-7.01 (m, 1H), 6.33 (br s, 1H), 6.27 (s, 1H), 5.98 (d, J=1.2 Hz, 1H), 5.94 (d, J=1.3 Hz, 1H), 5.68 (s, 1H), 4.50 (dd, J=11.2, 3.2 Hz, 1H), 4.13-4.11 (m, 2H), 4.05 (d, J=2.2 Hz, 1H), 3.90 (dd, J=11.2, 3.4 Hz, 1H), 3.57 (br s, 3H), 3.33 (d, J=7.8 Hz, 1H), 3.17 (dt, J=11.9, 2.7 Hz, 1H), 2.95 (dd, J=17.9, 8.2 Hz, 1H), 2.83 (d, J=14.4 Hz, 1H), 2.63 (d, J=17.8 Hz, 1H), 2.34 (s, 3H), 2.24 (s, 3H), 2.03 (s, 3H), 1.87-1.81 (m, 1H), 1.81 (br s, 3H); FTIR (neat) 3375 (m br), 2933 (m br), 2873 (w), 1733 (m br), 1601 (m), 1533 (m), 1501 (m), 1445 (m), 1417 (m), 1371 (m), 1314 (m), 1299 (m), 1266 (m), 1214 (s), 1155 (m), 1145 (m), 1109 (m), 1086 (m), 1070 (m), 1029 (m), 1007 (m), 953 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C35H36O8N4Na: 663.2431, found 663.2417.
EXAMPLE 24
Figure USRE041614-20100831-C00057
Phthalimide (7) (0.3 mg, 0.00046 mmol) was dissolved in methylene chloride (0.2 mL) and 4-dimethylaminopyridine (0.6 mg, 0.0049 mmol) and acetic anhydride (1.0 μL, 0.010 mmol) were added to the solution. The reaction was stirred at 23° C. for 20 min and then purified by flash column chromatography (0.3 mL silica gel, gradient methylene chloride to ethyl acetate) to afford Compound 23 (0.3 mg, 94%). Rf0.19 (1:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.72-7.65 (m, 4H), 6.78 (s, 1H), 5.70 (s, 1H), 5.40 (br s, 1H), 4.25-4.23 (m, 2H), 3.72-3.63 (m, 2H), 3.63-3.50 (m, 4H), 3.38 (d, J=7.6 Hz, 1H), 3.19 (d, J=12.2 Hz, 1H), 3.05 (dd, J=18.1, 8.0 Hz, 1H), 2.72 (d, J=18.0 Hz, 1H), 2.62 (d, J=14.6 Hz, 1H), 2.33 (s, 3H), 2.31 (s, 3H), 2.25 (s, 3H), 2.23 (s, 3H), 2.00 (s, 3H), 1.78-1.63 (m, 1H); FTIR (neat) 2931 (m br), 2850 (w), 1769 (s), 1713 (s), 1494 (w), 1431 (m br), 1394 (m), 1369 (m), 1238 (m), 1194 (s), 1100 (m), 1075 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C38H36O9Na: 715.2380, found 715.2360.
EXAMPLE 25
Figure USRE041614-20100831-C00058
Phthalimide (7) (0.7 mg, 0.0011 mmol) was dissolved in methylene chloride (0.2 mL) and N,N-diisopropylethylamine (1.0 μL, 0.0058 mmol) and N-chlorosuccinimide (0.66 mg, 0.0049 mmol) were added to the solution. The reaction was stirred at 23° C. for 28 hr and passed through a small plug of silica gel with ethyl acetate. The mixture was concentrated in vacuo and the residue was purified by preparative thin layer chromatography (10% ethyl acetate-methylene chloride, three elutions) to afford Compound 24 (0.5 mg, 68%). Rf0.19 (10% ethyl acetate-methylene chloride); 1H NMR (500 MHz, CDCl3) δ7.72-7.70 (m, 2H), 7.65-7.63 (m, 2H), 5.70 (s, 1H), 5.56 (s, 1H), 5.39 (br s, 1H), 4.28 (d, J=2.2 Hz, 1H), 4.25 (t, J=5.4 Hz, 1H), 4.07 (s, 1H), 3.66 (d, J=4.9 Hz, 2H), 3.60 (s, 3H), 3.46 (d, J=8.3 Hz, 1H), 3.22 (d, J=11.7 Hz, 1H), 2.96 (dd, J=18.7, 8.0 Hz, 1H), 2.76 (d, J=15.8 Hz, 1H), 2.70 (d, J=18.6 Hz, 1H), 2.30 (s, 3H), 2.28 (s, 6H), 1.99 (s, 3H), 1.67 (t, J=12.4 Hz, 1H); FTIR (neat) 3407 (m br), 2936 (m br), 2854 (w), 1764 (m), 1716 (s), 1466 (m), 1452 (m), 1431 (m), 1408 (m), 1395 (m), 1369 (m), 1315 (w), 1273 (w), 1235 (m), 1197 (m), 1146 (w), 1102 (m), 1086 (m), 1074 (m), 1031 (m), 1003 (w), 947 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H33O8N4ClNa: 707.1885, found 707.1888.
EXAMPLE 26
Figure USRE041614-20100831-C00059
Phthalimide (7) (0.5 mg, 0.00077 mmol) was dissolved in a 0.0056 M solution of N-bromosuccinimide in methylene chloride (0.14 mL, 0.00079 mmol). The reaction was stirred at 23° C. for 40 min and was then quenched into a saturated solution of sodium thiosulfate (10 mL). The mixture was extracted with ethyl acetate (10 mL) and the organic layers were washed with water (2×20 mL) and saturated aqueous sodium chloride (20 mL), dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by preparative thin layer chromatography (10% ethyl acetate-methylene chloride, two elutions) to afford Compound 25 (0.5 mg, 89%). Rf0.16 (10% ethyl acetate-methylene chloride); 1H NMR (500 MHz, CDCl3) δ7.73-7.71 (m, 2H), 7.65-7.63 (m ,2H), 5.68 (s, 1H), 5.60 (s, 1H), 5.36 (br s, 1H), 4.28 (s, 1H), 4.25 (t, J=5.3 Hz, 1H), 4.08 (s, 1H), 3.65 (d, J=5.0 Hz, 2H), 3.61 (s, 3H), 3.46 (d, J=8.1 Hz, 1H), 3.22 (d, J=11.5 Hz, 1H), 2.94 (dd, J=18.7, 8.1 Hz, 1H), 2.76 (d, J=15.7 Hz, 1H), 2.69 (d, J=18.4 Hz, 1H), 2.35 (s, 3H), 2.28 (s, 6H), 1.99 (s, 3H), 1.69-1.63 (m, 1H); FTIR (neat) 3412 (m br), 2935 (m br), 2856 (w), 1764 (m), 1717 (s), 1461 (m), 1449 (m), 1431 (m), 1405 (m), 1395 (m), 1369 (m), 1196 (m), 1101 (m), 1075 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H33O8N4BrNa: 751.1379, found 751.1399.
EXAMPLE 27
Figure USRE041614-20100831-C00060
Phthalimide (7) (0.5 mg, 0.00077 mmol) was dissolved in 3:2 acetonitrile-water (0.25 mL). Silver nitrate (4.0 mg, 0.024 mmol) was added as a solid and the solution was stirred at 23° C. for 11 hr. The reaction was quenched by stirring with a 1:1 mixture of saturated aqueous sodium chloride and saturated aqueous sodium bicarbonate (0.5 mL) for 15 min. The mixture was poured into a 1:1 mixture of saturated aqueous sodium chloride and saturated aqueous sodium bicarbonate (2 mL) and extracted with methylene chloride (3×4 mL), dried over sodium sulfate, filtered through Celite and concentrated in vacuo to afford Compound 26 (0.3 mg, 60%). 1H NMR (400 MHz, CDCl3) δ7.72-7.70 (m, 2H), 7.66-7.62 (m, 2H), 6.43 (s, 1H), 5.58 (s, 1H), 5.59 (s, 1H), 5.15 (br s, 1H), 4.65-4.58 (m, 2H), 4.01 (d, J=10.6 Hz, 1H), 3.93 (s, 1H), 3.69-3.50 (m, 5H), 3.26 (d, J=11.8 Hz, 1H), 3.15 (d, J=7.2 Hz, 1H), 2.92 (dd, J=18.2, 8.4 Hz, 1H), 2.71 (d, J=15.2 Hz, 1H), 2.58 (d, J=17.5 Hz, 1H), 2.30 (s, 3H), 2.25 (s, 3H), 2.24 (s, 3H), 1.96 (s, 3H), 1.76-1.67 (m, 1H); FTIR (neat) 3436 (m br), 2960 (m br), 2929 (m br), 2855 (w), 1762 (m), 1716 (s), 1499 (m), 1459 (m), 1432 (m), 1394 (m), 1367 (m), 1293 (w), 1262 (w), 1233 (m), 1199 (m), 1149 (w), 1103 (m), 1073 (m), 1030 (m), 1007 (m), 946 (w) cm−1.
EXAMPLE 28
Figure USRE041614-20100831-C00061
Phthalimide (4) (3.6 mg, 0.0052 mmol) was azeotropically dried with toluene (2×2 mL) and dissolved in THF (0.5 mL). The mixture was cooled to −78° C. in a dry ice-acetone bath and a 1.0 M solution of L-Selectride in THF (10 μL, 0.010 mmol) was added drop-wise. The reaction was warmed to 23° C. slowly over 5 hr and was quenched with 2 drops of 5% acetic acid in water. After stirring at 23° C. for 30 min the reaction was concentrated in vacuo, dissolved in ethyl acetate, passed through a short pad a silica gel using ethyl acetate and concentrated in vacuo. This residue was dissolved in methylene chloride (0.8 mL) and to this solution was added acetic acid (5.0 □L, 0.088 mmol), PdCl2(PPh3)2 (1.0 mg, 1.4 μmol) and tributyltin hydride (4.0 μL, 0.015 mmol). Bubbling was observed and the reaction changes from a yellow to a dark orange color. After stirring at 23° C. for 10 min the reaction was charged with 4-dimethylaminopyridine (13.2 mg, 0.11 mmol) and acetic anhydride (10 μL, 0.10 mmol). The reaction was stirred at 23° C. for 5 min and purified by flash column chromatography (1 mL silica gel, gradient methylene chloride to 2:1 ethyl acetate-hexane). This residue was further purified by preparative thin layer chromatography (2:1 ethyl acetate-hexane) to afford a 55:45 mixture of isomers (3.2 mg, 84%).
The mixture of compounds (1.9 mg, 0.0026 mmol) was dissolved in methylene chloride (0.5 mL) and the solution treated with triethylsilane (28 μL, 0.175 mmol) and trifluoroacetic acid (10 μL, 0.129 mmol). After stirring at 23° C. for 10 min the reaction was concentrated in vacuo and purified twice by preparative thin layer chromatography (2:1 ethyl acetate-hexane and 5% methanol-methylene chloride) to afford a residue (0.9 mg, 51%).
This material (0.8 mg, 0.0012 mmol) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.2 mL) and the solution was stirred at 23° C. for 11 hr. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed from repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by flash column chromatography (1.0 mL silica gel, methylene chloride to 1:1 to 2:1 ethyl acetate-hexane) to afford Compound 27 (0.8 mg, 100%). Rf0.20 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.75-7.73 (m, 1H), 7.39-7.35 (m, 2H), 7.08 (d, J=7.2 Hz, 1H), 6.14 (s, 1H), 5.97 (s, 1H), 5.92 (s, 1H), 5.44 (s, 1H), 4.36 (d, J=1.9 Hz, 1H), 4.24 (d, J=4.4 Hz, 1H), 3.97 (d, J=2.3 Hz, 1H), 3.59 (s, 3H), 3.55-3.34 (m, 5H), 3.24 (d, J=11.6 Hz, 1H), 2.89-2.84 (m, 2H), 2.77 (d, J=15.6 Hz, 1H), 2.29 (s, 3H), 2.27 (s, 3H), 2.05 (s, 3H), 1.96 (s, 3H), 1.62-1.60 (m, 1H); FTIR (neat) 3379 (m br), 2932 (m br), 2857 (w), 1759 (s), 1682 (s), 1619 (w), 1588 (w), 1499 (w), 1455 (m), 1434 (m), 1416 (m), 1370 (m), 1327 (w), 1303 (w), 1234 (m), 1199 (s), 1148 (w), 1105 (m), 1085 (m), 1076 (m), 1030 (w), 1000 (w), 956 (w), 913 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H36O7N4Na: 659.2482, found 659.2488.
EXAMPLE 29
Figure USRE041614-20100831-C00062
Phthalimide (4) (3.6 mg, 0.0052 mmol) was azeotropically dried with toluene (2×2 mL) and dissolved in THF (0.5 mL). The mixture was cooled to −78° C. in a dry ice-acetone bath and a 1.0 M solution of L-Selectride in THF (10 μL, 0.010 mmol) was added dropwise. The reaction was warmed to 23° C. slowly over 5 hr and was quenched with 2 drops of 5% acetic acid in water. After stirring at 23° C. for 30 min the reaction was concentrated in vacuo, dissolved in ethyl acetate, passed through a short pad a silica gel using ethyl acetate and concentrated in vacuo. This residue was dissolved in methylene chloride (0.8 mL) and to this solution was added acetic acid (5.0 μL, 0.088 mmol), PdCl2(PPh3)2 (1.0 mg, 1.4 μmol) and tributyltin hydride (4.0 μL, 0.015 mmol). Bubbling was observed and the reaction changes from a yellow to a dark orange color. After stirring at 23° C. for 10 min the reaction was charged with 4-dimethylaminopyridine (13.2 mg, 0.11 mmol) and acetic anhydride (10 μL, 0.10 mmol). The reaction was stirred at 23° C. for 5 min and purified by flash column chromatography (1 mL silica gel, gradient methylene chloride to 2:1 ethyl acetate-hexane). This residue was further purified by preparative thin layer chromatography (2:1 ethyl acetate-hexane) to afford a 55:45 mixture of isomers (3.2 mg, 84%).
This material (3.0 mg, 0.004 mmol) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.2 mL) and the solution was stirred at 23° C. for 11 h. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by flash column chromatography (1.0 mL silica gel, methylene chloride to 2:1 ethyl acetate-hexane) and then purified by preparative thin layer chromatography (2:1 ethyl acetate-hexane) to afford isomerically pure (28) (1.2 mg, 46%). Rf0.18 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.67 (d, J=6.7 Hz, 1H), 7.44-7.40 (m, 2H), 7.26-7.24 (m, 1H), 6.09 (s, 1H), 5.98 (s, 1H), 5.96 (s, 1H), 5.42 (s, 1H), 4.95 (d, J=9.5 Hz, 1H), 4.34 (d, J=2.3 Hz, 1H), 4.31 (br s, 1H), 4.24 (d, J=4.5 Hz, 1H), 3.96 (d, J=2.2 Hz, 1H), 3.64 (d, J=14.2 Hz, 1H), 3.39-3.12 (m, 6H), 3.25 (dt, J=11.7, 2.7 Hz, 1H), 2.85 (dd, J=18.1, 7.4 Hz, 1H), 2.77 (d, J=17.9 Hz, 1H), 2.29 (s, 3H), 2.26 (s, 3H), 2.07 (s, 3H), 1.91 (s, 3H), 1.79 (d, J=9.6 Hz, 1H); FTIR (neat) 3375 (m br), 2933 (m br), 2857 (w), 2256 (w), 1758 (m), 1686 (s), 1499 (w), 1435 (s), 1370 (m), 1326 (m), 1299 (s), 1292 (s), 1234 (s), 1199 (s), 1147 (w), 1124 (m), 1105 (m), 1084 (s), 1075 (s), 1031 (m), 1008 (m), 953 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H36O8N4Na: 675.2431, found 675.2439.
EXAMPLE 30
Figure USRE041614-20100831-C00063
Alcohol (4) (14.3 mg, 0.025 mmol) was azeotropically dried with toluene (2×1 mL) in vacuo. The residue was dissolved in methylene chloride (0.5 mL) and to this solution was added N,N-diisopropylethylamine (9.0 μL, 0.052 mmol), 4-dimethylaminopyridine (9.4 mg, 0.077 mmol) and p-toluenesulfonic anhydride (29.0 mg, 0.089 mmol). The reaction was stirred at 23° C. for 13 hr and was then quenched into a half-saturated solution of aqueous sodium bicarbonate (10 mL). The mixture was extracted with methylene chloride (3×10 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by flash column chromatography (10 mL silica gel, 1:1 ethyl acetate-hexane) to afford Compound 29 (12.6 mg, 69% yield). Rf0.32 (1:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ7.62 (d, J=8.1 Hz, 2H), 7.27 (d, J=7.9 Hz, 2H), 6.70 (s, 1H), 6.10-6.07 (m, 1H), 5.79 (s, 2H), 5.40 (d, J=15.8 Hz, 1H), 5.27 (d, J=10.3 Hz, 1H), 5.13-5.09 (m, 2H), 4.20-4.10 (m, 5H), 3.95 (dd, J=9, 3, 3.0 Hz, 1H), 3.74 (s, 3H), 3.57 (s, 3H), 3.51 (t, J=9.8 Hz, 1H), 3.30 (d, J=8.0 Hz, 1H), 3.22 (d, J=13.6 Hz, 2H), 3.02 (d, J=17.9, 7.9 Hz, 1H), 2.65 (d, J=17.9 Hz, 1H), 2.44 (s, 3H), 2.31 (s, 3H), 2.26 (s, 3H), 2.10 (s, 3H), 1.78 (dd, J=15.7, 12.2 Hz, 1H); 13C NMR (125 MHz, CDCl3) δ148.6, 148.3, 148.2, 144.7, 144.4, 139.3, 133.7, 132.9, 130.5, 129.7, 127.8, 125.3, 123.7, 121.4, 118.0, 117.7, 113.0, 110.2, 101.2, 99.3, 74.3, 73.5, 61.6, 59.7, 57.7, 57.5, 57.1, 55.9, 55.6, 41.5, 26.2, 25.4, 21.6, 15.8, 9.3; FTIR (neat) 2935 (m br), 2256 (w), 1738 (w), 1600 (w), 1484 (w), 1449 (m), 1402 (w), 1364 (m), 1342 (m), 1295 (w), 1268 (w), 1232 (m), 1189 (m), 1177 (s), 1158 (m), 1096 (s), 1066 (m), 1021 (m), 998 (m), 970 (m), 962 (m), 930 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C38H43O9N3SNa: 740.2618, found 740.2649; [α]D 23+78.7° (c 0.97, methylene chloride).
EXAMPLE 31
Figure USRE041614-20100831-C00064
Tosylate (29) (14.0 mg, 0.020 mmol) was dissolved in DMF (0.5 mL). Lithium azide (7.7 mg, 0.16 mmol) was added and the reaction was placed in a 70° C. oil bath for 20 min. The reaction was cooled to room temperature, diluted with 1:1 ethyl acetate-hexane (20 mL) and washed with water (3×20 mL) and saturated aqueous sodium chloride (20 mL). The organic layers were dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by preparative thin layer chromatography (2:1 diethyl ether-hexane, two elutions) to afford Compound 30 (8.4 mg, 73% yield). Rf0.43 (1:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ6.61 (s, 1H), 6.15-6.08 (m, 1H), 5.94 (d, J=1.4 Hz, 1H), 5.87 (d, J=1.4 Hz, 1H), 5.41 (dq, J=17.2, 1.5 Hz, 1H), 5.28 (ddd, J=11.5, 1.6, 1.1 Hz, 1H), 5.14 (d, J=5.9 Hz, 1H), 5.11 (d, J=5.9 Hz, 1H), 4.24-4.12 (m, 4H), 4.01 (dd, J=7.1, 2.9 Hz, 1H), 3.73 (s, 3H), 3.58 (s, 3H), 3.40 (dd, J=12.1, 3.0 Hz, 1H), 3.35 (d, J=7.6 Hz, 1H), 3.27 (dd, J=6.7, 2.6 Hz, 1H), 3.24 (d, J=2.6 Hz, 1H), 3.12-3.02 (m, 2H), 2.63 (d, J=17.9 Hz, 1H), 2.33 (s, 3H), 2.22 (s, 3H), 2.13 (s, 3H), 1.89 (dd, J=15.8, 12.1 Hz, 1H); 13C NMR (100 MHz, CDCl3) δ148.6, 148.4, 148.3, 144.4, 139.0, 133.7, 130.6, 130.2, 125.4, 123.7, 121.3, 118.1, 117.7, 112.7, 112.4, 101.2, 99.3, 74.2, 61.3, 59.7, 57.7, 57.1, 56.9, 56.5, 55.5, 41.5, 26.3, 25.6, 15.7, 9.3; FTIR (neat) 2934 (s br), 2857 (m), 2105 (s), 1725 (w), 1650 (w), 1613 (w), 1581 (w), 1484 (m), 1444 (s), 1342 (m), 1323 (m), 1302 (m), 1269 (m), 1232 (m), 1158 (m), 1104 (s), 1096 (m), 1078 (m), 1024 (m), 999 (s), 977 (m), 928 (m), 914 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C31H36O6N6Na: 611.2594, found 611.2613; [α]D 23+71.0° (c 0.73, methylene chloride).
EXAMPLE 32
Figure USRE041614-20100831-C00065
Azide (30) was dissolved in nitrogen degassed methanol (0.5 mL). To the solution was added triethylamine (21 μL, 0.15 mmol) and dithiothreitol (24.0 mg, 0.16 mmol). The reaction was stirred at 23° C. for 17 hr and then concentrated in vacuo. The residue was dissolved in ethyl acetate (20 mL) and washed with water (20 mL). The aqueous layer was extracted with ethyl acetate (20 mL) and the combined organic layers were washed with saturated aqueous sodium chloride (20 mL), dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by flash column chromatography (5 mL silica gel, gradient diethyl ether to 5% methanol-methylene chloride) to afford Compound 31 (4.9 mg, 59% yield). Rf0.10 (5% methanol-methylene chloride); 1H NMR (400 MHz, CDCl3) δ6.70 (s, 1H), 6.14-6.10 (m, 1H), 5.93 (d, J=1.4 Hz, 1H), 5.86 (d, J=1.4 Hz, 1H), 5.40 (dd, J=17.1, 1.5 Hz, 1H), 5.27 (dd, J=10.3, 1.4 Hz, 1H), 5.12 (s, 2H), 4.23 (d, J=2.0 Hz, 1H), 4.22-4.18 (m, 1H), 4.14 (dd, J=12.1, 5.8 Hz, 1H), 3.99 (d, J=2.5 Hz, 1H), 3.91 (s, 1H), 3.71 (s, 3H), 3.59 (s, 3H), 3.37 (d, J=7.3 Hz, 1H), 3.28 (dt, J=11.7, 2.7 Hz, 1H), 3.21 (dd, J=15.8, 2.7 Hz, 1H), 3.09 (dd, J=17.9, 8.0 Hz, 1H), 2.76 (dd, J=17.7, 2.5 Hz, 1H), 2.71 (dd, J=13.7, 3.3 Hz, 1H), 2.49 (d, J=17.9 Hz, 1H), 2.35 (s, 3H), 2.21 (s, 3H), 2.13 (s, 3H), 1.81 (dd, J=15.7, 11.8 Hz, 1H), 1.34 (br s, 2H); 13C NMR (100 MHz, CDCl3) δ148.6, 148.43, 148.37, 144.5, 138.8, 133.8, 130.8, 130.0, 125.1, 124.0, 121.3, 117.9, 117.6, 113.7, 112.2, 101.1, 99.3, 74.1, 59.9, 59.8, 58.9, 57.7, 57.1, 56.3, 55.3, 44.2, 41.7, 26.5, 25.7, 15.8, 9.3; FTIR (neat) 3100 (v/v br), 2934 (m br), 2860 (w), 1484 (w), 1446 (m), 1432 (m), 1385 (m), 1376 (w), 1341 (m), 1323 (w), 1299 (w), 1269 (m), 1233 (w), 1158 (m), 1103 (s), 1075 (m), 1064 (m), 1042 (m), 1023 (s), 998 (m), 977 (m), 964 (m), 927 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C31H38O6N4Na: 585.2689, found 585.2693; [α]D 23+86.8° (c 0.41, methylene chloride).
EXAMPLE 33
Figure USRE041614-20100831-C00066
Tosylate (29) (1.0 mg, 0.0014 mmol) was dissolved in a saturated solution of potassium 4-pyridinedicarboimide (0.2 mL, −30 equiv.). After stirring at 23° C. for 4 hr the reaction was diluted with 1:1 ethyl acetate-hexane (10 mL) and washed with water (10 mL). The aqueous was extracted with 1:1 ethyl acetate-hexane (10 mL) and the combined organic layers were washed with saturated aqueous sodium chloride (10 mL), dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by flash column chromatography (1.0 mL silica gel, gradient methylene chloride to 2:1 diethyl ether-hexane) to afford a residue (0.9 mg, 94%).
This material was dissolved in methylene chloride (0.3 mL) and to this solution was added acetic acid (1.0 μL, 0.018 mmol), PdCl2(PPh3)2 (0.5 mg, 0.6 μmol) and tributyltin hydride (2.5 μL, 0.0093 mmol). Bubbling was observed and the reaction changes from a yellow to a dark orange color. After stirring at 23° C. for 10 min the reaction was charged with 4-dimethylaminopyridine (2.5 mg, 0.020 mmol) and acetic anhydride (2.5 μL, 0.025 mmol). The reaction was stirred at 23° C. for 5 min and purified by preparative thin layer chromatography (2:1 ethyl acetate-hexane) to afford a residue (0.9 mg, 100 %).
This material was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.2 mL) and the solution was stirred at 23° C. for 11 hr. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by flash column chromatography (1.0 mL silica gel, gradient methylene chloride to 2:1 diethyl ether-hexane to ethyl acetate) and then by preparative thin layer chromatography (2:1 ethyl acetate-hexane, two elutions) to afford Compound 32 (0.7 mg, 83%). Rf0.14 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, C6D6) δ8.75 (s, 1H), 8.27 (d, J=4.8 Hz, 1H), 6.91 (s, 1H), 6.39 (s, 1H), 5.31 (br s, 1H), 5.24 (s, 1H), 5.01 (br s, 1H), 4.47 (d, J=3.7 Hz, 1H), 3.93 (d, J=2.2 Hz, 1H), 3.81-3.76 (m, 2H), 3.62 (dd, J=13.9, 5.5 Hz, 1H), 3.37 (d, J=11.4 Hz, 1H), 2.99 (s, 3H), 2.86 (d, J=4.2 Hz, 1H), 2.79 (d, J=17.1 Hz, 1H), 2.62-2.60 (m, 2H), 2.16 (s, 3H), 2.08 (s, 3H), 2.01 (s, 3H), 1.76 (s, 3H), 1.59 (m, 1H); FTIR (neat) 3431 (w br), 2935 (m br), 2856 (w), 1761 (m), 1723 (s), 1615 (w), 1499 (m), 1434 (m), 1388 (m), 1369 (w), 1327 (w), 1301 (w), 1294 (m), 1268 (m), 1234 (m), 1197 (m), 1145 (m), 1137 (m), 1100 (m), 1074 (m), 1030 (m), 1007 (m), 997 (m), 947 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C35H33O8N5Na: 674.2227, found 674.2237.
EXAMPLE 34
Figure USRE041614-20100831-C00067
Amine (31) (0.6 mg, 0.0011 mmol) was dissolved in methylene chloride (0.5 mL). To this mixture was added 4-dimethylaminopyridine (0.5 mg, 0.0041 mmol) and α,α-dibromoxylene (0.5 mg, 0.0019 mmol). After stirring at 23° C. for 3 hr the reaction was purified by flash column chromatography (0.6 mL silica gel, gradient methylene chloride to 1:1 ethyl acetate-hexane) to afford a film (0.5 mg, 71%).
This residue was dissolved in methylene chloride (0.5 mL) and to this solution was added acetic acid (0.5 μL, 0.0088 mmol), PdCl2(PPh3)2 (0.02 mg, 0.04 μmol) and tributyltin hydride (1.0 μL, 0.0037 mmol). Bubbling was observed and the reaction changes from a yellow to a dark orange color. After stirring at 23° C. for 10 min the reaction was charged with 4-dimethylaminopyridine (2.0 mg, 0.016 mmol) and acetic anhydride (1.0 μL, 0.010 mmol). The reaction was stirred at 23° C. for 5 min and purified by preparative thin layer chromatography (1:1 ethyl acetate-hexane, three elutions).
This material was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 0.5 mL) and the solution was stirred at 23° C. for 11 hr. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by preparative thin layer chromatography (2:1 ethyl acetate-hexane, two elutions) to afford Compound 33 (0.2 mg, 43% over two steps). Rf0.43 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ7.20-7.19 (m, 4H), 6.46 (s, 1H), 5.95 (d, J=1.2 Hz, 1H), 5.91 (d, J=1.3 Hz, 1H), 5.64 (s, 1H), 4.94 (br s, 1H), 4.10 (d, J=8.3 Hz, 1H), 4.07 (s, 1H), 3.99 (d, J=11.9 Hz, 2H), 3.87 (d, J=11.4 Hz, 2H), 3.77 (s, 3H), 3.26 (d, J=12.0 Hz, 1H), 3.20 (d, J=7.2 Hz, 1H), 2.91 (dd, J=17.7, 8.1 Hz, 1H), 2.87 (d, J=12.6 Hz, 1H), 2.79 (d, J=16.6 Hz, 1H), 2.75-2.71 (m, 1H), 2.59 (d, J=17.8 Hz, 1H), 2.33 (s, 3H), 2.30 (s, 3H), 2.26 (s, 3H), 2.00 (s, 3H), 1.91 (dd, J=16.1, 11.2 Hz, 1H); FTIR (neat) 3406 (w br), 2927 (s), 2854 (m), 1762 (m), 1719 (m), 1459 (m), 1500 (w), 1432 (m), 1370 (m), 1325 (w), 1294 (w), 1233 (m), 1199 (s), 1144 (m), 1105 (m), 1085 (m), 1074 (m), 1029 (m), 1006 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H39O6N4: 623.2870, found 623.2878.
EXAMPLE 35
Figure USRE041614-20100831-C00068
Amine (31) (0.6 mg, 0.0011 mmol) was dissolved in methylene chloride (0.5 mL). To this mixture was added 4-dimethylaminopyridine (0.5 mg, 0.0041 mmol), pyruvic acid (0.5 μL, 0.0072 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.5 mg, 0.0026 mmol). After stirring at 23° C. for 3 hr the reaction was purified by flash column chromatography (0.6 mL silica gel, gradient methylene chloride to 1:1 ethyl acetate-hexane) to afford a film (0.5 mg, 73%).
This residue was dissolved in methylene chloride (0.5 mL) and to this solution was added acetic acid (0.5 μL, 0.0088 mmol), PdCl2(PPh3)2 (0.02 mg, 0.04 μmol) and tributyltin hydride (1.5 μL, 0.0056 mmol). Bubbling was observed and the reaction changes from a yellow to a dark orange color. After stirring at 23° C. for 10 min the reaction was charged with 4-dimethylaminopyridine (2.0 mg, 0.016 mmol) and acetic anhydride (1.0 μL, 0.010 mmol). The reaction was stirred at 23° C. for 5 min and purified by preparative thin layer chromatography (1:1 ethyl acetate-hexane, three elutions).
This material was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 0.5 mL) and the solution was stirred at 23° C. for 11 hr. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by preparative thin layer chromatography (2:1 ethyl acetate-hexane, two elutions) to afford Compound 34 (0.3 mg, 64% over two steps). Rf0.30 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ6.45 (s, 1H), 6.42 (br s, 1H), 5.99 (d, J=1.1 Hz, 1H), 5.93 (d, J=1.2 Hz, 1H), 5.66 (s, 1H), 4.08-4.06 (m, 2H), 4.01 (d, J=2.4 Hz, 1H), 3.79 (s, 3H), 3.46-3.42 (m, 2H), 3.35 (d, J=7.8 Hz, 1H), 3.25 (d, J=11.7 Hz, 1H), 3.03 (dd, J=18.1, 8.5 Hz, 1H), 2.79 (d, J=14.1 Hz, 1H), 2.56 (d, J=17.7 Hz, 1H), 2.31 (s, 3H), 2.30 (s, 3H), 2.26 (s, 3H), 2.21 (s, 3H), 2.00 (s, 3H), 1.77 (t, J=13.6 Hz, 1H); FTIR (neat) 3382 (m br), 2929 (m br), 2854 (w), 1761 (m), 1735 (m), 1721 (m), 1687 (s), 1519 (w), 1509 (w), 1500 (w), 1458 (m), 1417 (m), 1368 (m), 1325 (w), 1294 (w), 1233 (m), 1199 (s), 1155 (m), 1108 (m), 1087 (m), 1030 (w), 1006 (w), 956 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C31H34O8N4Na: 613.2274, found 613.2195.
EXAMPLE 36
Figure USRE041614-20100831-C00069
TABLE 2
General procedure for the EDC-HCl Coupling of Carboxylic
to phenol (5).
Cmpd Side Chain Stoichiometry Coupling MOM Removal
Entry # Reagent (equiv.) Yield (%) Yield (%)
1 35
Figure USRE041614-20100831-C00070
 		70 100 82
2 36
Figure USRE041614-20100831-C00071
 		4.5 94 100
3 37
Figure USRE041614-20100831-C00072
 		5.9 95 96
4 38
Figure USRE041614-20100831-C00073
 		4.9 93 96
5 39
Figure USRE041614-20100831-C00074
 		6.8 55 100
Phenol (5) (1.0 mg, 0.0015 mmol) was dissolved in a 0.0126 M solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine in methylene chloride (0.5 mL, 0.0064 mmol of each). The carboxylic acid was added and the reaction was stirred at 23° C. for 30 min and then quenched into a saturated solution of aqueous sodium bicarbonate (2 mL). The mixture was extracted with methylene chloride (2×5 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by flash column chromatography (1 mL silica gel, gradient methylene chloride to 1:1 to 2:1 ethyl acetate-hexane) to afford the corresponding phenolic esters. This material was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL) and the solution was stirred at 23° C. for 11 h. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by flash column chromatography (1 mL silica gel, gradient methylene chloride to 2:1 ethyl acetate-hexane to ethyl acetate)8 to afford the desired product.
8 Entry 5 was purified using 5% methanol-methylene chloride as the eluent.
EXAMPLE 37
Compound 35—Rf0.30 (1:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) α 77.2-7.69 (m, 2H), 7.67-7.64 (m, 2H), 6.39 (s, 1H), 5.69 (s, 1H), 5.60 (s, 1H), 5.38 (br s, 1H), 4.36-4.22 (m, 4H), 4.00 (d, J=1.9 Hz, 1H), 3.67 (d, J=5.2 Hz, 2H), 3.61 (s, 3H), 3.55 (s, 3H), 3.36 (d, J=8.3 Hz, 1H), 3.19 (d, J=12.0 Hz, 1H), 3.02 (dd, J=18.0, 8.3 Hz, 1H), 2.73 (dd, J=15.6, 2.3 Hz, 1H), 2.68 (d, J=18.3 Hz, 1H), 2.29 (s, 3H), 2.21 (s, 3H), 2.00 (s, 3H), 1.73 (dd, J=15.4, 12.3 Hz, 1H); FTIR (neat) 3420 (w), 2933 (m), 2872 (w), 2854 (w), 1774 (m), 1716 (s), 1432 (m), 1394 (m), 1118 (m), 1105 (m), 1071 (m), 1231 (m), 1162 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C37H36O9N4Na: 703.2380, found 703.2373.
EXAMPLE 38
Compound 36—Rf 0.52 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) α 7.74-7.71 (m, 2H), 7.68-7.65 (m, 2H), 6.39 (s, 1H), 5.65 (s, 1H), 5.57 (s, 1H), 5.27 (br s, 1H), 4.25-4.23 (m, 2H), 4.06 (s, 1H), 3.65 (s, 3H), 3.63-3.61 (m, 2H), 3.38 (d, J=6.2 Hz, 1H), 3.22 (d, J=12.0 Hz, 1H), 3.03 (dd, J=17.9, 8.0 Hz, 1H), 2.77 (d, J=14.1 Hz, 1H), 2.66 (d, J=18.1 Hz, 1H), 2.60 (q, J=7.6 Hz, 2H), 2.31 (s, 3H), 2.22 (s, 3H), 1.97 (s, 3H), 1.79-1.72 (m, 1H), 1.31 (t, J=7.6 Hz, 3H); FTIR (neat) 3450 (m br), 2979 (w), 2935 (m br), 1771 (w), 1759 (m), 1716 (s), 1460 (m), 1432 (m), 1418 (m), 1394 (m), 1234 (m), 1191 (m), 1144 (m), 1102 (m), 1089 (m), 1070 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C37H36O8N4Na: 687.2431, found 687.2421.
EXAMPLE 39
Compound 37—Rf 0.60 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) α 7.73-7.69 (m, 2H), 7.67-7.64 (m 2), 7.47-7.36 (m, 5H), 6.35 (s, 1H), 5.63 (s, 1H), 5.25 (br s, 1H), 5.13 (br s, 1H), 4.22-4.19 (m, 2H), 3.94 (d, J=2.3 Hz, 1H), 3.86 (s, 2H), 3.60-3.58 (m, 2H), 3.53 (br s, 3H), 3.33 (d, J=8.2 Hz, 1H), 3.16 (d, J=12.0 Hz, 1H), 3.00 (dd, J=17.9, 8.1 Hz, 1H), 2.67 (d, J=15.6 Hz, 1H), 2.60 (d, J=18.1 Hz, 1H), 2.27 (s, 3H), 2.19 (s, 3H), 1.89 (s, 3H), 1.68-1.61 (m, 1H); FTIR (neat) 3429 (m br), 2932 (m br), 2856 (w), 1761 (w), 1735 (m), 1715 (s), 1498 (w), 1456 (m), 1432 (m), 1395 (m), 1324 (m), 1296 (w), 1233 (m), 1191 (w), 1120 (ml, 1104 (m), 1083 (m), 1071 (m), 1029 (w), 1004 (w), 946 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C42H38O8N4Na: 749.2587, found 749.2577.
EXAMPLE 40
Compound 38—Rf 0.61 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) α 7.74-7.72 (m, 2H), 7.67-7.65 (m, 2H), 7.37-7.24 (m, 5H), 6.38 (s, 1H), 5.63 (s, 1H), 5.50 (s, 1H), 5.25 (br s, 1H), 4.25-4.21 (m, 2H), 4.01 (d, J=2.1 Hz, 1H), 3.64 (s, 3H), 3.62-3.60 (m, 2H), 3.34 (d, J=8.1 Hz, 1H), 3.20 (d, J=1.02 Hz, 1H), 3.13-3.08 (m, 2H), 3.02 (dd, J=18.1, 8.0 Hz, 1H), 2.92-2.88 (m, 2H), 2.76 (d, J=14.8 Hz, 1H), 2.63 (d, J=18.0 Hz, 1H), 2.29 (s, 3H), 2.22 (s, 3H), 1.87 (s, 3H), 1.76 (dd, J=15.2, 12.0 Hz, 1H); FTIR (neat) 3427 (m br), 2934 (m br), 2858 (w), 1758 (m), 1716 (s), 1455 (m), 1432 (m), 1395 (m), 1350 (w), 1316 (w), 1256 (m), 1132 (m), 1104 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C43H40O8N4Na: 763.2744, found 763.2755.
EXAMPLE 41
Compound 39—Rf 0.17 (4:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) α 7.73-7.71 (m, 2H), 7.67-7.64 (m, 2H), 6.36 (s, 1H), 6.00 (s, 1H), 5.65 (d, J=1.4 Hz, 1H), 5.27 (d, J=1.4 Hz, 1H), 4.70 (br s, 1H), 4.27 (d, J=2.3 Hz, 1H), 4.22 (t, J=6.2 Hz, 1H), 4.11 (s, 1H), 4.01 (s, 1H), 3.68-3.62 (m, 5H), 3.35 (d, J=7.3 Hz, 1H), 3.17 (d, J=11.8 Hz, 1H), 3.03 (dd, J=18.0, 8.2 Hz, 1H), 2.85 (d, J=14.3 Hz, 1H), 2.63 (d, J=17.9 Hz, 1H), 2.32 (s, 3H), 2.20 (s, 3H), 2.14 (s, 3H), 1.99 (s, 3H), 1.70 (dd, J=15.4, 12.2 Hz, 1H); FTIR (neat) 3382 (m br), 2934 (m br), 1774 (m), 1716 (s), 1673 (m), 1538 (w), 1500 (w), 1459 (m), 1432 (m), 1419 (m), 1396 (m), 1377 (m), 1293 (w), 1234 (m), 1153 (m), 1133 (m), 1103 (m), 1072 (m), 1031 (w), 944 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C38H37O9N5Na: 730.2489, found 730.2492.
EXAMPLE 42
Figure USRE041614-20100831-C00075
TABLE 3
General Procedure for the Alkylation of Phenol (5).
Stoichi- MOM
Cmpd Alkylating ometry Alkylation Removal
Entry # Reagent (equiv.) Yield (%) Yield (%)
1 40
Figure USRE041614-20100831-C00076
 		5.7 92 90
2 41
Figure USRE041614-20100831-C00077
 		5.7 48 100
3 42
Figure USRE041614-20100831-C00078
 		5.9 75 100
Phenol (5) (1.0 mg, 0.0015 mmol) was azeotropically dried with toluene (2×1 mL) in vacuo and dissolved in DMF (0.1 mL). Cesium carbonate (3.0 mg, 0.0092 mmol) was gently flame dried in vacuo, cooled and added as a solid to the reaction mixture. The alkylation agent was added via syringe and the solution was stirred at 23° C. for 4 hr and then quenched into a saturated solution of aqueous sodium bicarbonate (2 mL). The mixture was extracted with 1:1 ethyl acetate-hexane (2×5 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by preparative thin layer chromatography (2:1 ethyl acetate-hexane, one elution). This material was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL) and the solution was stirred at 23° C. for 11 hr. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by flash column chromatography (1 mL silica gel, gradient methylene chloride to 2:1 ethyl acetate-hexane) to afford the desire product.
EXAMPLE 43
Compound 40—Rf 0.43 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ7.73-7.72 (m, 2H), 7.67-7.66 (m, 2H), 6.40 (s, 1H), 5.67 (s, 1H), 5.56 (d, J=1.5 Hz, 1H), 5.10 (d, J=1.5 Hz, 1H), 4.22-4.19 (m, 2H), 4.10 (d, J=2.0 Hz, 1H), 3.70 (s, 3H), 3.61 (s, 3H), 3.59-3.51 (m, 2H), 3.35 (d, J=8.3 Hz, 1H), 3.24-3.19 (m, 2H), 3.05 (dd, J=18.1, 8.2 Hz, 1H), 2.63 (d, J=17.9 Hz, 1H), 2.31 (s, 3H), 2.25 (s, 3H), 2.09 (s, 3H), 1.85 (dd, J=15.7, 12.2 Hz, 1H); FTIR (neat) 3428 (w br), 2935 (m br), 1774 (m), 1716 (s), 1619 (w), 1588 (w), 1499 (w), 1432 (m), 1423 (m), 1396 (m), 1324 (m), 1301 (m), 1266 (m), 1233 (m), 1191 (m), 1145 (w), 1101 (m), 1066 (m), 1028 (m), 998 (m), 948 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C35H34O7N4Na: 645.2325, found 645.2325.
EXAMPLE 44
Compound 41—Rf 0.45 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.72-7.65 (m, 4H), 6.41 (s, 1H), 5.65 (s, 1H), 5.57 (d, J=1.4 Hz, 1H), 5.11 (d, J=1.4 Hz, 1H), 4.22-4.19 (m, 2H), 4.08 (s, 1H), 3.73 (q, J=7.0 Hz, 2H), 3.68 (s, 3H), 3.62-3.53 (m, 2H), 3.35 (d, J=8.2 Hz, 1H), 3.25-3.17 (m, 2H), 3.05 (dd, J=18.2, 8.2 Hz, 1H), 2.65 (d, J=18.0 Hz, 1H), 2.31 (s, 3H), 2.24 (s, 3H), 2.09 (s, 3H), 1.80 (dd, J=15.3, 11.6 Hz, 1H), 1.35 (t, J=7.0 Hz, 3H); FTIR (neat) 3412 (m br), 2930 (m br), 1773 (m), 1716 (s), 1619 (w), 1588 (w), 1500 (w), 1455 (m), 1395 (m), 1386 (m), 1370 (m), 1265 (m), 1233 (m), 1145 (m), 1101 (m), 1066 (m), 1028 (m), 1006 (m), 950 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H36O7N4Na: 659.2482, found 659.2477.
EXAMPLE 45
Compound 42—Rf0.53 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.69-7.63 (m, 4H), 6.43 (s, 1H), 5.62-5.61 (m, 2H), 5.18 (d, J=1.5 Hz, 1H), 4.23-4.20 (m, 2H), 4.07 (d, J=2.5 Hz, 1H), 3.93 (m, 1H), 3.62 (m, 5H), 3.35 (d, J=8.4 Hz, 1H), 3.25 (dd, J=15.4, 2.0 Hz, 1H), 3.14 (d, J=12.1 Hz, 1H), 3.03 (dd, J=17.9, 8.1 Hz, 1H), 2.69 (d, J=17.9 Hz, 1H), 2.29 (s, 3H), 2.24 (s, 3H), 2.08 (s, 3H), 1.72 (dd, J=15.3, 11.9 Hz, 1H), 1.24 (d, J=4.4 Hz, 3H), 1.22 (d, J=4.4 Hz, 3H); FTIR (neat) 3435 (m br), 2973 (m br), 2933 (m br), 1773 (m), 1716 (s), 1619 (w), 1588 (w), 1500 (w), 1461 (m), 1432 (m), 1395 (m), 1384 (m), 1233 (m), 1144 (m), 1100 (m), 1075 (m), 1064 (m), 1029 (m), 1006 (w), 998 (w), 947 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C37H38O7N4Na: 673.2638, found 673.2663.
EXAMPLE 46
Figure USRE041614-20100831-C00079
Phenol (5) (1.0 mg, 0.0015 mmol) was dissolved in methylene chloride (0.5 mL) to this solution were added 4-dimethylaminopyridine (0.8 mg, 0.0066 mmol) and n-butyric anhydride (1.0 μL, 0.0061 mmol). The reaction was stirred at 23° C. for 15 min and then quenched into a saturated solution of aqueous sodium bicarbonate (2 mL). The mixture was extracted with methylene chloride (2×5 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by flash column chromatography (1 mL silica gel, gradient methylene chloride to 1:1 to 2:1 ethyl acetate-hexane) to afford a residue (0.9 mg, 83%). This material was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL) and the solution was stirred at 23° C. for 11 hr. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by flash column chromatography (1 mL silica gel, gradient methylene chloride to 2:1 ethyl acetate-hexane) to afford Compound 43 (0.7 mg, 93%). Rf0.56 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.74-7.71 (m, 2H), 7.68-7.65 (m, 2H), 6.38 (s, 1H), 5.64 (s, 1H), 5.55 (s, 1H), 5.25 (s, 1H), 4.27-4.21 (m, 2H), 4.02 (d, J=2.3 Hz, 1H), 3.66 (s, 3H), 3.62-3.60 (m, 2H), 3.34 (d, J=7.7 Hz, 1H), 3.20 (d, J=11.9 Hz, 1H), 3.02 (dd, J=18.0, 8.0 Hz, 1H), 2.78 (d, J=15.3 Hz, 1H), 2.63 (d, J=18.0 Hz, 1H), 2.55 (dt, J=2.5, 7.3 Hz, 2H), 2.28 (s, 3H), 2.23 (s, 3H), 1.97 (s, 3H), 1.82 (q, J=7.4 Hz, 2H), 1.78-1.72 (m, 1H), 1.08 (t, J=7.4 Hz, 3H); FTIR (neat) 3433 (m br), 2934 (m br), 2876 (w), 1758 (m), 1716 (s), 1499 (w), 1459 (m), 1432 (m), 1395 (m), 1328 (w), 1296 (w), 1234 (m), 1190 (w), 1172 (m), 1146 (m), 1102 (m), 1072 (m), 1029 (w), 1005 (w), 998 (w), 947 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C38H38O8N4Na: 701.2587, found 701.2581.
EXAMPLE 47
Figure USRE041614-20100831-C00080
Phenol (5) (1.0 mg, 0.0015 mmol) was dissolved in methylene chloride (0.5 mL) to this solution were added 4-dimethylaminopyridine (0.8 mg, 0.0066 mmol) and methanesulfonyl chloride (0.5 μL, 0.0065 mmol). The reaction was stirred at 23° C. for 15 min and then quenched into a saturated solution of aqueous sodium bicarbonate (2 mL). The mixture was extracted with methylene chloride (2×5 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by flash column chromatography (1 mL silica gel, gradient methylene chloride to 1:1 to 2:1 ethyl acetate-hexane) to afford a residue (0.9 mg, 82%). This material was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL) and the solution was stirred at 23° C. for 11 h. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by flash column chromatography (1 mL silica gel, gradient methylene chloride to 2:1 ethyl acetate-hexane) to afford Compound 44 (0.8 mg, 100%). Rf0.45 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.68-7.62 (m, 4H), 6.43 (s, 1H), 5.76 (d, J=1.5 Hz, 1H), 5.60 (s, 1H), 5.47 (d, J=1.5 Hz, 1H), 4.25-4.22 (m, 2H), 4.06 (d, J=2.2 Hz, 1H), 3.73 (dd, J=14.0, 6.9 Hz, 1H), 3.67 (dd, J=14.0, 3.3 Hz, 1H), 3.55 (s, 3H), 3.37 (d, J=8.0 Hz, 1H), 3.20-3.13 (m, 5H), 3.03 (dd, J=18.1, 8.1 Hz, 1H), 2.73 (d, J=18.0 Hz, 1H), 2.30 (s, 3H), 2.22 (s, 3H), 2.21 (s, 3H), 1.85 (dd, J=16.0, 12.0 Hz, 1H); FTIR (neat) 3464 (m br), 2936 (m br), 2855 (w), 1774 (w), 1716 (s), 1499 (w), 1461 (m), 1433 (m), 1394 (m), 1366 (m), 1295 (w), 1234 (w), 1178 (m), 1145 (w), 1101 (m), 1081 (w), 1070 (m), 1058 (m), 1030 (w), 996 (w), 971 (w), 948 (w), 890 (m), 808 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C35H34O9N4SNa: 709.1944, found 709.1956.
EXAMPLE 48
Figure USRE041614-20100831-C00081
Methoxymethyl ether (4) (0.5 mg, 0.00072 mmol) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL) and the solution was stirred at 23° C. for 11 h. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by flash column chromatography (0.5 mL silica gel, gradient methylene chloride to 2:1 ethyl acetate-hexane) to afford Compound 45 (0.4 mg, 85%). Rf0.53 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.73-7.65 (m, 4H), 6.41 (s, 1H), 6.18-6.02 (m, 1H), 5.61 (s, 1H), 5.58 (d, J=1.5 Hz, 1H), 5.38 (dd, J=17.2, 1.5 Hz, 1H), 5.24 (d, J=10.3 Hz, 1H), 5.13 (d, J=1.4 Hz, 1H), 4.23-4.13 (m, 3H), 4.08 (s, 1H), 3.97 (dd, J=7.5, 5.9 Hz, 1H), 3.68 (s, 3H), 3.59-3.52 (m, 2H), 3.35 (d, J=8.2 Hz, 1H), 3.24 (dd, J=17.3, 2.1 Hz, 1H), 3.18 (d, J=11.4 Hz, 1H), 3.04 (dd, J=17.3, 8.0 Hz, 1H), 2.64 (d, J=18.2 Hz, 1H), 2.30 (s, 3H), 2.24 (s, 3H), 2.09 (s, 3H), 1.80 (dd, J=15.2, 11.7 Hz, 1H); FTIR (neat) 3413 (w br), 2931 (m br), 2856 (w), 1775 (w), 1713 (s), 1463 (m), 1431 (m), 1394 (m), 1300 (w), 1269 (w), 1231 (w), 1144 (w), 1100 (m), 1063 (w), 1031 (w), 994 (w), 950 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C37H36O7N4Na: 671.2482, found 671.2498.
EXAMPLE 49
Figure USRE041614-20100831-C00082
The methoxymethyl ether (5) (5.0 mg, 0.0077 mmol) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.2 mL) and the solution. was stirred at 23° C. for 11 h under an atmosphere of oxygen. The reaction mixture was diluted with toluene (5 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×5 mL). The residue was purified by preparative TLC (2:1 ethyl acetate-hexane) to afford Compound 46 (2.3 mg, 50%). Rf0.38 (5% methanol-methylene chloride, 1H NMR (400 MHz, C6D6) δ7.07-7.05 (m, 2H), 6.66-6.64 (m, 2H), 6.31 (s, 1H), 5.10 (s, 1H), 4.07 (br s, 1H), 4.06 (d, J=2.0 Hz, 1H), 3.81 (d, J=2.3 Hz, 1H), 3.74 (dd, J=14.5, 1.8 Hz, 1H), 3.67 (dd, J=14.6, 4.5 Hz, 1H), 3.53 (br s, 1H), 3.19-3.14 (m, 2H), 2.86 (d, J=7.9 Hz, 1H), 2.79 (d, J=18.3 Hz, 1H), 2.72 (s, 3H), 2.60 (dd, J=18.1, 7.7 Hz, 1H), 2.29-2.17 (m, 1H), 2.10 (s, 3H), 1.94 (s, 3H), 1.88 (s, 3H); FTIR (neat) 3399 (m br), 2928 (m br), 2855 (m), 1773 (w), 1713 (s), 1657 (m), 1644 (m), 1631 (m), 1436 (m), 1416 (m), 1396 (m), 1382 (m), 1378 (m), 1360 (m), 1334 (m), 1303 (m), 1245 (m), 1234 (w), 1172 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C33H30O7N4Na: 617.2012, found 617.2036.
EXAMPLE 50
Figure USRE041614-20100831-C00083
The hydroxyquinone (46) (2.3 mg, 0.0038 mmol) was dissolved in methylene chloride (3 mL). A dilute diazomethane solution in diethyl ether was added in small portions while monitoring the reaction by TLC analysis. Upon complete conversion to the product, acetic acid (50 μL) was added to quench the reaction. Purification via preparative TLC (1:1 ethyl acetate-hexane) afforded pure (47) (1.0 mg, 42%). Rf0.33 (1:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ7.59 (m, 4H), 6.43 (s, 1H), 5.48 (s, 1H), 4.39 (d, J=2.3 Hz, 1H), 4.11 (br s, 1H), 4.07 (s, 3H), 4.07-4.03 (m, 1H), 4.01 (d, J=2.0 Hz, 1H), 3.81 (dd, J=14.5, 1.3 Hz, 1H), 3.43-3.39 (m, 1H), 3.40 (s, 3H), 3.03 (dt, J=11.1, 2.7 Hz, 1H), 2.98-2.93 (m, 3H), 2.30 (s, 3H), 2.12 (s, 3H), 1.96 (s, 3H); FTIR (neat) 3459 (m br), 2934 (m br), 2855 (m), 1773 (m), 1713 (s), 1659 (s), 1641 (m), 1622 (m), 1499 (m), 1437 (s), 1396 (m), 1362 (m), 1302 (m), 1282 (m), 1267 (m), 1239 (s), 1163 (m), 1149 (m), 1138 (m), 1104 (m), 1087 (m), 1061 (m), 997 (m), 969 (m), 921 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C34H32O7N4Na: 631.2169, found 631.2183.
EXAMPLE 51
Figure USRE041614-20100831-C00084
The methoxymethyl ether (5) (0.6 mg, 0.00092 mmol) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL)) and the solution was stirred at 23° C. for 7 h. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by flash column chromatography (0.4 mL silica gel, gradient methylene chloride to 2:1 ethyl acetate-hexane) to afford Compound 48 (0.4 mg, 71%). Rf0.37 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ7.45-7.73 (m, 2H), 7.69-7.66 (m, 2H), 6.39 (s, 1H), 5.71 (s, 1H), 5.51 (s, 1H), 5.02 (s, 1H), 4.28-4.17 (m, 3H), 4.15-4.07 (m, 1H), 3.74 (s, 3H), 3.59-3.49 (m, 2H), 3.35 (d, J=8.0 Hz, 1H), 3.23 (d, J=11.9 Hz, 1H), 3.11-3.02 (m, 2H), 2.62 (d, J=18.0 Hz, 1H), 2.31 (s, 3H), 2.24 (s, 3H), 2.07 (s, 3H), 1.86 (dd, J=14.9, 11.8 Hz, 1H); FTIR (neat) 3464 (m br), 2934 (m br), 1772 (m), 1713 (s), 1460 (m br), 1433 (m br), 1416 (m br), 1367 (w), 1324 (w), 1234 (m), 1102 (m), 1075 (w), 1061 (w), 1028 (w), 1006 (w) cm−1; HRMS (FAB), [m+H]/z calc'd for C34H33O7N4: 609.2349, found 609.2341.
EXAMPLE 52
Figure USRE041614-20100831-C00085
Phthalimide (5) (5.4 mg, 0.0083 mmol) was dissolved in ethanol (0.3 mL) and hydrazine (26 μL, 0.829 mmol) was added. The vessel was sealed and heated to 80° C. for 2 h. The reaction was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (2×1 mL). The residue was purified by flash column chromatography (0.5 mL silica gel, gradient methylene chloride to 5% methanol-ethyl acetate) to afford Compound 49 (4.3 mg, 100%). Rf0.18 (5% methanol-ethyl acetate); 1H NMR (400 MHz, CDCl3) δ6.66 (s, 1H), 5.88 (s, 1H), 5.80 (s, 1H), 5.34 (d, J=6.1 Hz, 1H), 5.19 (d, J=6.0 Hz, 1H), 4.23 (d, J=2.3 Hz, 1H), 4.01 (d, J=2.6 Hz, 1H), 3.94 (s, 1H), 3.70 (s, 3H), 3.69 (s, 3H), 3.37 (d, J=7.7 Hz, 1H), 3.33 (d, J=9.0 Hz, 1H), 3.10-3.04 (m, 2H), 2.83 (d, J=13.8 Hz, 1H), 2.74 (dd, J=13.7, 2.5 Hz, 1H), 2.49 (d, J=18.0 Hz, 1H), 2.34 (s, 3H), 2.20 (s, 3H), 2.08 (s, 3H), 1.79 (dd, J=15.0, 11.9 Hz, 1H); 13C NMR (100 MHz, CDCl3) δ149.1, 147.6, 145.5, 144.6, 135.9, 131.0, 130.2, 124.8, 122.9, 117.8, 113.1, 112.6, 106.1, 100.7, 99.8, 59.8, 59.5, 58.8, 57.7, 56.7, 55.6, 55.3, 43.6, 41.7, 26.2, 25.7, 15.7, 8.8; FTIR (neat) 3346 (w br), 3000 (w v br), 2935 (s br), 1446 (s br), 1419 (m), 1401 (m), 1327 (m), 1152 (m), 1101 (s), 1075 (m), 1060 (m), 998 (m), 975 (m) cm−1; HRMS (FAB), [m+H]/z calc'd for C28H35O6N4: 523.2557, found 523.2552; [α]D 23=−16.5° (c 0.20, methylene chloride).
EXAMPLE 53
Figure USRE041614-20100831-C00086
Amine (49) (0.9 mg, 0.0017 mmol) and acid EJM-III-124C (0.5 mg, 0.0026 mmol) were azeotropically tried with toluene (2×1 mL) and then dissolved in methylene chloride (0. 1 mL). 1,3-Dicyclohexylcarbodiimide (0.7 mg, 0.0034 mmol) was added to the solution which was stirred at 23° C. for 30 min. White precipitate was observed and the reaction was quenched into saturated aqueous sodium bicarbonate solution (7 mL). The aqueous layer was extracted with methylene chloride (2×7 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by flash column chromatography (1.0 mL silica gel, gradient 2:1 ethyl acetate-hexane to ethyl acetate) to afford Compound 50 (0.5 mg, 50%). Rf0.16 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ6.91 (s, 1H), 6.83 (d, J=8.2 Hz, 2H), 6.77 (s, 1H), 6.67 (d, J=8.4 Hz, 2H), 5.83 (s, 1H), 5.74 (s, 1H), 5.58 (s, 1H), 5.33 (d, J=6.3 Hz, 1H), 5.17 (d, J=6.0 Hz, 1H), 4.90 (m, 1H), 4.23 (s, 1H), 4.14 (s, 1H), 3.98 (s, 1H), 3.70 (s, 3H), 3.69 (s, 3H), 3.57-3.46 (m, 2H), 3.38 (d, J=7.5 Hz, 1H), 3.25 (d, J=11.5 Hz, 1H), 3.15 (d, J=15.8 Hz, 1H), 3.02-2.98 (m, 1H), 2.85 (d, J=15.8 Hz, 1H), 2.69 (d, J=18.0 Hz, 1H), 2.32 (s, 3H), 2.28 (s, 3H), 2.24 (s, 3H), 2.06 (s, 3H), 1.65 (m, 1H); FTIR (neat) 3400 (w br), 2924 (s br), 2853 (s), 1763 (m), 1753 (m), 1745 (m), 1737 (m), 1461 (m), 1452 (w), 1440 (w), 1350 (w), 1234 (w), 1216 (m), 1197 (m), 1160 (w) cm−1; HRMS (FAB), [m+H]/z calc'd for C38H43O9N4: 699.3030, found 699.3005.
EXAMPLE 54
Figure USRE041614-20100831-C00087
The methoxymethyl ether (50) (1.3 mg, 0.0019 mmol) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL) and the solution was stirred at 23□C for 10 h. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by flash column chromatography (0.3 mL silica gel, gradient 2:1 ethyl acetate-hexane to ethyl acetate) to afford Compound 51 (0.5 mg, 42%). Rf0.19 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ6.74 (d, J=8.4 Hz, 2H), 6.56 (s, 1H), 6.48 (d, J=8.4 Hz, 2H), 5.89 (s, 1H), 5.79 (s, 1H), 5.75 (s, 1H), 4.92 (s, 1H), 4.81 (s, 1H), 4.12 (s, 1H), 4.01 (d, J=13.3 Hz, 2H), 3.81 (m, 4H), 3.36 (d, J=7.1 Hz, 1H), 3.27 (d, J=13.0 Hz, 1H), 3.21 (d, J=15.5 Hz, 1H), 3.16 (d, J=12.0 Hz, 1H), 3.02 (dd, J=17.9, 8.3 Hz, 1H), 2.89 (d, J=15.6 Hz, 1H), 2.72 (d, J=15.4 Hz, 1H), 2.49 (d, J=18.4 Hz, 1H), 2.39 (s, 3H), 2.34 (s, 3H), 2.30 (s, 3H), 2.06 (s, 3H), 1.11 (dd, J=15.3, 11.3 Hz, 1H); FTIR (neat) 3388 (s br), 2931 (s br), 1754 (m), 1657 (m), 1506 (m), 1460 (m br), 1434 (m br), 1369 (m), 1233 (s), 1194 (s), 1099 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H38O8N4Na: 677.2587, found 677.2573.
EXAMPLE 55
Figure USRE041614-20100831-C00088
p-Hydroxyphenylacetic acid (52) (100 mg, 0.657 mmol) was dissolved in DMF (3.0 mL). tert-Butyldimethylsily chloride (222 mg, 1.47 mmol) and N,N-diisopropylethylamine (0.285 mL, 1.64 mmol were added to the solution which was stirred at 23° C. for 3 h. Water (1 mL) was added and after 15 min the reaction mixture was poured into 5% aqueous acetic acid (25 mL) and extracted with ethyl acetate (2×25 mL). The organic layers were then washed with water (2×20 mL) and saturated aqueous sodium chloride (20 mL), dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by flash column chromatography (120 mL silica gel, gradient 1:4 to 1:1 ethyl acetate-hexane, 0.1% acetic acid) to afford Compound 53 (125 mg, 72%). Rf0.52 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.14 (d, J=8.5 Hz, 2H), 6.80 (d, J=8.5 Hz, 2H), 3.57 (s, 2H), 0.98 (s, 9H), 0.19 (s, 6H); 13C NMR (100 MHz, CDCl3) δ178.3, 154.9, 130.3, 125.8, 120.1, 40.3, 25.6, 18.2, −4.4; FTIR (neat) 3122 (w br), 2957 (m), 2931 (m), 2897 (m), 2888 (m), 2859 (m), 1712 (s), 1611 (w), 1512 (s), 1472 (w), 1464 (w), 1409 (w), 1263 (s), 1171 (w), 917 (s), 840 (m), 826 (m), 803,(m) cm−1; HRMS (EI), [m+] calc'd for C14H22O3Si: 266.1338, found 266.1331.
EXAMPLE 56
Figure USRE041614-20100831-C00089
Amine (49) (2.0 mg, 0.0038 mmol) and acid (53) (1.3 mg, 0.0049 mmol) were azeotropically dried with toluene (2×1 mL) and then dissolved in methylene chloride (0.2 mL). 1,3-Dicyclohexylcarbodiimide (1.0 mg, 0.0049 mmol) was added to the solution which was stirred at 23° C. for 30 min. White precipitate was observed and the reaction was quenched into saturated solution of aqueous sodium bicarbonate (5 mL). The aqueous layer was extracted with methylene chloride (2×5 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by flash column chromatography (1.7 mL silica gel, gradient 1:4 ethyl acetate-hexane to ethyl acetate) to afford Compound 54 (1.4 mg, 47%). Rf0.39 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ6.76 (s, 1H), 6.55 (d, J=8.4 Hz, 2H), 6.47 (d, J=8.3 Hz, 2H), 5.82, (s, 1H), 5.74 (s, 1H), 5.53 (s, 1H), 5.35 (d, J=6.1 Hz, 1H), 5.18 (d, J=6.1 Hz, 1H), 4.89 (br s, 1H), 4.20 (s, 1H), 4.15 (d, J=2.3 Hz, 1H), 3.97 (s, 1H), 3.70 (s, 6H), 3.67-3.58 (m, 1H), 3.44-3.36 (m, 2H), 3.23 (d, J=11.4 Hz, 1H), 3.11 (d, J=16.1 Hz, 1H), 3.02-2.96 (m, 2H), 2.83 (d, J=16.1 Hz, 1H), 2.70 (d, J=18.2 Hz, 1H), 2.31 (s, 3H), 2.23 (s, 3H), 2.07 (s, 3H), 1.67-1.57 (m, 1H), 0.96 (s, 9H), 0.14 (s, 3H), 0.13 (s, 3H); FTIR (neat) 3396 (w br), 2930 (m br), 2857 (m), 1656 (m br), 1651 (w), 1509 (s), 1462 (m br), 1257 (s), 1157 (m), 1097 (s), 1060 (m), 915 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C42H54O8N4SiNa: 793.3609, found 793.3624.
EXAMPLE 57
Figure USRE041614-20100831-C00090
Phenol (54) (1.1 mg, 0.0015 mmol) was dissolved in methylene chloride (0.2 mL). 4-Dimethylaminopyridine (0.4 mg, 0.0032 mmol) and acetic anhydride (0.5 μL, 0.0053 mmol) were added to the solution which was stirred at 23° C. for 1 h. The reaction mixture was concentrated in vacuo. The residue was purified by flash column chromatography (0.3 mL silica gel, gradient 1:1 ethyl acetate-hexane to ethyl acetate) to afford Compound 55 (1.2 mg, 100%). Rf0.48 (2:1 ethyl acetate-hexane); 1H NMR (500 MHz, CDCl3) δ6.78 (s, 1H), 6.75-6.65 (m, 4H), 5.89 (d, J=1.3 Hz, 1H), 5.84 (d, J=1.3 Hz, 1H), 5.16 (d, J=5.7 Hz, 1H), 5.06 (d, J=5.7 Hz, 1H), 5.03 (br s, 1H), 4.18 (s, 2H), 3.96 (s, 1H), 3.71 (s, 3H), 3.62 (m, 1H), 3.58 (s, 3H), 3.39-3.32 (m, 2H), 3.25 (d, J=13.0 Hz, 1H), 3.01 (dd, J=18.2, 8.2 Hz, 1H), 2.95 (d, J=15.4 Hz, 1H), 2.83 (d, J=15.7 Hz, 1H), 2.78 (m, 1H), 2.74 (d, J=18.2 Hz, 1H), 2.35 (s, 3H), 2.34 (s, 3H), 2.17 (s, 3H), 1.99 (s, 3H), 1.78 (dd, J=15.3, 11.6 Hz, 1H), 0.94 (s, 9H), 0.12 (s, 3H), 0.11 (s, 3H); FTIR (neat) 3404 (w br), 2932 (s br), 2858 (m), 1761 (m), 1673 (m), 1509 (s), 1442 (m br), 1368 (m), 1259 (s), 1201 (s), 1159 (m), 1089 (m), 916 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C44H56O9N4SiNa: 835.3714, found 835.3699.
EXAMPLE 58
Figure USRE041614-20100831-C00091
The methoxymethyl ether (55) (1.2 mg, 0.0015 mmol) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL) and the solution was stirred at 23° C. for 10 h. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by flash column chromatography (0.3 mL silica gel, ethyl acetate) to afford Compound 56 (0.4 mg, 44%). Rf0.13 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ6.56-6.48 (m, 5H), 5.92 (d, J=1.3 Hz, 1H), 5.85 (d, J=1.3 Hz, 1H), 5.74 (s, 1H), 5.36 (s, 1H), 4.88 (br s, 1H), 4.14-4.08 (m, 2H), 3.98 (s, 1H), 3.78 (s, 3H), 3.68-3.43 (m, 2H), 3.36 (d, J=7.4 Hz, 1H), 3.22 (d, J=11.9 Hz, 1H), 3.07-2.93 (m, 3H), 2.82-2.73 (m, 1H), 2.66 (d, J=16.3 Hz, 1H), 2.38 (s, 3H), 2.29 (s, 3H), 2.34 (s, 3H), 1.98 (s, 3H), 1.74-1.65 (m, 1H); FTIR (neat) 3403 (s br), 2929 (s br), 2856 (m), 1756 (m), 1656 (m), 1513 (s), 1450 (m), 1369 (m), 1233 (s), 1201 (s), 1086 (m) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H38O8N4Na: 677.2587, found 677.2587.
EXAMPLE 59
Figure USRE041614-20100831-C00092
Compound 7 1.0 mg, 0.00154 mmol) was dissolved in THF (1.0 mL). Salcomine (0.1 mg, 0.00031 mmol) was added as a solid to make an orange solution. The vial was secured inside a bomb reactor and the vessel was purged with oxygen ten times and filled to 50 psi. (˜3 bar). The solution was stirred at 23° C. for 3 h. The reaction concentrated in vacuo, passed through a small pad of silica get eluting with ethyl acetate and purified by preparative thin layer chromatography (1:1 acetate-hexane, 2×) to afford Compound 57 (0.9 mg, 90%). Rf0.51 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.70 (m, 4H), 5.85 (s, 1H), 5.72 (br s, 1H), 4.32 (t, J=5.1 Hz, 1H), 4.02 (d, J=2.2 Hz, 1H), 3.90 (s, 1H), 3.80 (s, 3H), 3.73 (d, J=5.1 Hz, 2H), 3.34 (d, J=7.3 Hz, 1H), 3.15 (d, J=12.1 Hz, 1H), 2.73 (dd, J=20.8, 7.3 Hz, 1H), 2.46 (dd, J=15.2, 2.0 Hz, 1H), 2.32 (s, 1H), 2.27 (s, 3H), 2.26 (s, 3H), 2.03 (s, 3H), 2.01 (s, 3H), 1.50-1.43 (m, 1H); FTIR (neat) 2938 (w), 2898 (w), 2851 (w), 1764 (m), 1716 (s), 1649 (m), 1616 (m), 1432 (m), 1393 (m), 1371 (m), 1308 (m), 1232 (m), 1197 (m), 1146 (m), 1101 (m), 1084 (w), 1029 (w), 946 (w), 906 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H32O9N4Na: 687.2067, found 687.2061.
TABLE 4
General Procedure for the Coupling of Alcohol (9) with
Dicarboximides.
Mitsunobu MOM
Coupling Removal
Entry Dicarboximide Yield Yield
Ex. 60
Figure USRE041614-20100831-C00093
 		57% No. 58 73% No. 59
Ex. 61
Figure USRE041614-20100831-C00094
 		60% No. 60 100% No. 61
EXAMPLE 60
Compound 59. The amount of Compound 9 was increased to 2.3 mg for this reaction. Preparative thin layer chromatography of the first step was done using 2:1 diethyl ether-hexane. Rf0.54 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ6.44 (s, 1H), 5.86 (s, 1H), 5.83 (br s, 1H), 5.63 (s, 1H), 4.20 (d, J=2.3 Hz, 1H), 4.11 (dd, J=6.3, 3.2 Hz, 1H), 4.03 (d, J=2.3 Hz, 1H), 3.76 (s, 3H), 3.53 (br m, 2H), 3.35 (d, J=8.0 Hz, 1H), 3.16 (d, J=11.8 Hz, 1H), 2.99 (dd, J=18.2, 8.0 Hz, 1H), 2.76-2.72 (m, 2H), 2.30 (s, 3H), 2.28 (s, 3H), 2.26 (s, 3H), 2.21-2.08 (m, 4H), 2.00 (s, 3H), 1.64 (br m, 5H); FTIR (neat) 3445 (m br), 2935 (m br), 2864 (w), 1761 (m), 1708 (s), 1499 (w), 1433 (m), 1410 (m), 1373 (m), 1323 (w), 1299 (w), 1270 (w), 1231 (m), 1200 (m), 1145 (w), 1101 (m), 1075 (m), 1030 (w), 1005 (w), 935 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H38O8N4Na: 677.2587, found 677.2597.
EXAMPLE 61
Compound 62. The amount of Compound 9 was increased to 2.3 mg for this reaction. Preparative thin layer chromatography of the first step was done using 2:1 diethyl ether-hexane and again using 1:1 ethyl acetate-hexane. Rf0.50 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.80 (s, 1H), 7.72 (dd, J=7.9, 1.7 Hz, 1H), 7.55 (d, J=7.8 Hz, 1H), 6.40 (s, 1H), 5.74 (s, 1H), 5.58 (s, 1H), 5.51 (br s, 1H), 4.24-4.19 (m, 2H), 4.01 (d, J=2.5 Hz, 1H), 3.69 (d, J=4.3 Hz, 2H), 3.62 (s, 3H), 3.36 (d, J=8.2 Hz, 1H), 3.18 (d, J=11.8 Hz, 1H), 3.01 (dd, J=18.0, 8.1 Hz, 1H), 2.75 (d, J=15.5 Hz, 1H), 2.69 (d, J=18.0 Hz, 1H), 2.28 (s, 6H), 2.24 (s, 3H), 2.00 (s, 3H), 1.68-1.62 (m, 1H); FTIR (neat) 3431 (m br), 3059 (w), 2934 (m br), 2858 (w), 1763 (s), 1719 (s), 1610 (w), 1499 (w), 1430 (s), 1386 (s), 1324 (m), 1298 (m), 1269 (m), 1231 (s), 1199 (s), 1145 (m), 1102 (s), 1075 (s), 1030 (m), 1003 (m), 945 (w), 905 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H33O8N4BrNa: 751.1379, found 751.1367.
EXAMPLE 62
Figure USRE041614-20100831-C00095
Amine (31) (0.7 mg, 0.0012 mmol) was dissolved in methylene chloride (0.3 mL). To this mixture was added 4-dimethylaminopyridine (trace), triethylamine (1.0 μL, 0.0072 mmol) and benzoyl chloride (0.5 μL, 0.0044 mmol). After stirring at 23° C. for 1.25 h the reaction was purified by flash column chromatography (1.0 mL silica gel, gradient methylene chloride to 1: 1 ethyl acetate-hexane) to afford Compound 64 (0.8 mg, 96%).
This residue was dissolved in methylene chloride (0.3 mL) and to this solution was added acetic acid (1.0 μL, 0.018 mmol), PdCl2(PPh3)2 (0.5 mg, 0.7 μmol) and tributyltin hydride (1.5 μL, 0.0056 mmol). Bubbling was observed and the reaction changes from a yellow to a dark orange color. After stirring at 23° C. for 10 min the reaction was charged with 4-dimethylaminopyridine (3.0 mg, 0.025 mmol) and acetic anhydride (2.5 μL, 0.026 mmol). The reaction was stirred at 23° C. for 5 min and purified by flash column chromatography (1.0 mL silica gel, gradient methylene chloride to 2:1 ethyl acetate-hexane) to afford Compound 6.5 (0.7 mg, 88%).
The methoxymethyl ether (65) was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 v/v), 1.0 mL) and the solution was stirred at 23° C. for 11 h. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by preparative thin layer chromatography (2:1 diethyl ether-hexane, two elutions and 1:1 ethyl acetate-hexane) to afford Compound 63 (0.4 mg, 61%). Rf0.33 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.32-7.37 (m, 1H), 7.25-7.23 (m, 4H), 6.18 (br s, 1H), 5.99 (s, 1H), 5.93 (s, 1H), 5.69 (br s, 1H), 5.61 (br s, 1H), 4.17 (br s, 2H), 4.08 (s, 1H), 3.65 (br s, 5H), 3.37 (d, J=7.2 Hz, 1H), 3.30 (d, J=12.0 Hz, 1H), 2.93 (dd, J=17.9, 7.4 Hz, 1H), 2.84 (d, J=15.8 Hz, 1H), 2.65 (d, J=17.7 Hz, 1H), 2.34 (s, 3H), 2.28 (s, 3H), 2.02 (s, 3H), 1.93-1.86 (m, 1H), 1.86 (br s, 3H); FTIR (neat) 3411 (m br), 2929 (s br), 2858 (m), 1757 (m), 1716 (m), 1655 (m), 1580 (w), 1524 (m), 1487 (s), 1452 (m), 1371 (m), 1293 (m), 1268 (m), 1231 (m), 1201 (s), 1151 (m), 1085 (s), 1030 (w), 1006 (w), 954 (w), 909 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C35H36O7N4Na: 647.2482, found 647.2455.
EXAMPLE 63
Figure USRE041614-20100831-C00096
Phenol (5) (0.8 mg, 0.0012 mmol) was dissolved in THF (0.2 mL) and to this solution were added 4-dimethylaminopyridine (1.0 mg, 0.0082 mmol) and methylisocyanate (0.5 μL, 0.0085 mmol). The reaction was stirred at 23° C. for 19 h and then quenched into a saturated solution of aqueous sodium bicarbonate (2 mL). The mixture was extracted with 1:1 ethyl acetate-hexane (2×5 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by flash column chromatography (1 mL silica gel, gradient methylene chloride to 1:1 to 2:1 ethyl acetate-hexane) to afford Compound 67 (0.8 mg, 92%). This material was dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL) and the solution was stirred at 23° C. for 11 h. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by flash column chromatography (1 mL silica gel, gradient methylene chloride to 1:1 to 2:1 ethyl acetate-hexane) to afford Compound 66 (0.7 mg, 96%). Rf0.21 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.73-7.70 (m, 2H), 7.69-7.66 (m, 2H), 6.36 (s, 1H), 5.63 (d, J=5.9 Hz, 1H), 5.18 (s, 1H), 5.03 (m, 1H), 4.23-4.21 (m, 2H), 4.05 (d, J=2.2 Hz, 1H), 3.67 (s, 3H), 3.63-3.55 (m, 2H), 3.34 (d, J=7.6 Hz, 1H), 3.23 (d, J=11.7 Hz, 1H), 3.06-2.95 (m, 2H), 2.88 (d, J=4.7 Hz, 3H), 2.85 (d, J=4.6 Hz, 1H), 2.60 (d, J=18.0 Hz, 1H), 2.29 (s, 3H), 2.20 (s, 3H), 2.02 (s, 3H), 1.78 (dd, J=15.8, 12.0 Hz, 1H); FTIR (neat) 3390 (m br), 2936 (m br), 2828 (w), 1771 (w), 1712 (s), 1647 (m), 1622 (w), 1519 (m), 1458 (m), 1430 (m), 1399 (m), 1322 (w), 1308 (w), 1232 (s), 1192 (w), 1109 (s), 1070 (m), 1029 (w), 1005 (w), 943 (w), 884 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C36H35O8N5Na: 688.2383, found 688.2392.
EXAMPLE 64
Figure USRE041614-20100831-C00097
Compound 6 (2.7 mg, 0.0039 mmol) was dissolved in chloroform and the reaction was cooled to −20° C. To this solution was added solid m-chloro-peroxybenzoic acid (1.4 mg, −0.0058 mmol). The reaction was stirred at −20° C. for 20 min and the reaction was quenched with triethylamine (20 μL, 0.146 mmol). The reaction was warmed to 4° C. and trifluoroacetic anhydride (6 μL, 0.043 mmol) was added. After 10 min the reaction was poured into water (5 mL), extracted with methylene chloride (2×5 mL) and the organic layers were dried over sodium sulfate, decanted and concentrated in vacuo. The residue was purified by preparative thin layer chromatography (2:1 ethyl acetate-hexane, one elution) to afford starting material and two compounds related to desired product. These two compounds were dissolved in a mixture of trifluoroacetic acid-THF-water (4:1:1 (v/v), 1.0 mL) and the solution was stirred at 23° C. for 11 h. The reaction mixture was diluted with toluene (1 mL) and the solution was concentrated in vacuo. Additional volatiles were removed by repetitive in vacuo azeotropic concentration from toluene (3×1 mL). The residue was purified by flash column chromatography (1 mL silica gel, gradient methylene chloride to 1:1 to 2:1 ethyl acetate-hexane to ethyl acetate) to afford Compound 68 (0.6 mg, 25% over two steps). Rf0.21 (2:1 ethyl acetate-hexane); 1H NMR (400 MHz, CDCl3) δ7.74-7.71 (m, 2H), 7.69-7.65 (m, 2H), 6.41 (s, 1H), 5.66 (s, 1H), 5.63 (s, 1H), 5.29 (br s, 1H), 4.30 (d, J=2.8 Hz, 1H), 4.26 (d, J=1.7 Hz, 1H), 4.20 (t, J=5.8 Hz, 1H), 3.73-3.69 (m, 2H), 3.65 (s, 3H), 3.64-3.62 (m, 2H), 3.15-3.07 (m, 2H), 2.85 (d, J=18.0 Hz, 1H), 2.81 (d, J=16.2 Hz, 1H), 2.31 (s, 3H), 2.22 (s, 3H), 1.99 (s, 3H), 1.74 (t, J=13.6 Hz, 1H); FTIR (neat) 3430 (m br), 3330 (w br), 2929 (m br), 2857 (w), 1764 (m), 1714 (s), 1499 (w), 1458 (m), 1431 (s), 1394 (s), 1376 (m), 1324 (w), 1300 (w), 1270 (w), 1201 (s), 1105 (s), 1081 (m), 1024 (w), 1011 (w), 945 (w), 908 (w) cm−1; HRMS (FAB), [m+Na]/z calc'd for C35H32O8N4Na: 659.2118, found 659.2126.
Biological Results
The analogs described above were screened in vitro for anti-tumor activity.9 The human cancer cell lines used in these assays include A-549 (Lung), HCT116 (Colon), A375 (Melanoma) and PC-3 (Prostate) and values are reported as IC50 (ng/mL). The following tables summarize the activity of all the synthetic derivatives. An IC50 reading greater than 100 ng/mL is considered inactive in the screening tests conducted on the compounds of the present invention. Lower values represent higher activity.
9 Cancer cell antiproliferative assays were performed by Dr. Takashi Owa, a postdoctoral fellow in the Stuart L. Schreiber research group.
TABLE 5
Biological Data for B-Ring Substitution at Position X1
Figure USRE041614-20100831-C00098
Compound Analog A- HCT-
Entry # (X1 Group) 459 116 A375 PC-3
1 7
Figure USRE041614-20100831-C00099
 		0.62 0.25 0.11 0.36
2 10
Figure USRE041614-20100831-C00100
 		1.0 0.56 0.18 0.69
3 11
Figure USRE041614-20100831-C00101
 		2.4 1.1 0.67 2.1
4 12
Figure USRE041614-20100831-C00102
 		2.6 1.8 0.75 1.7
5 13
Figure USRE041614-20100831-C00103
 		3.6 1.7 0.40 1.7
6 22
Figure USRE041614-20100831-C00104
 		4.5 2.3 1.9 2.6
7 20
Figure USRE041614-20100831-C00105
 		14 2.1 0.79 2.0
8 14
Figure USRE041614-20100831-C00106
 		31 7.3 1.1 2.9
9 16
Figure USRE041614-20100831-C00107
 		58 6.8 1.6 9.2
10 18
Figure USRE041614-20100831-C00108
 		61 8.3 3.9 11
11 56
Figure USRE041614-20100831-C00109
 		>100 20 6.5 24
12 21
Figure USRE041614-20100831-C00110
 		>100 43 22 64
13 19
Figure USRE041614-20100831-C00111
 		>100 63 27 71
14 17
Figure USRE041614-20100831-C00112
 		>100 66 29 89
15 15
Figure USRE041614-20100831-C00113
 		>100 68 11 52
16 27
Figure USRE041614-20100831-C00114
 		7.2 2.2 0.71 1.7
17 28
Figure USRE041614-20100831-C00115
 		22 2.4 0.79 2.7
18 32
Figure USRE041614-20100831-C00116
 		11 2.1 0.36 37
TABLE 6
Biological Data for A-Ring Substitution at Position X2.
Figure USRE041614-20100831-C00117
Compound Analog A- HCT
Entry # (X2 Group) 549 116 A375 PC-3
1 7
Figure USRE041614-20100831-C00118
 		0.62 0.25 0.11 0.36
2 35
Figure USRE041614-20100831-C00119
 		1.1 0.61 0.22 0.63
3 44
Figure USRE041614-20100831-C00120
 		1.2 0.40 0.20 0.59
4 36
Figure USRE041614-20100831-C00121
 		1.4 0.80 0.34 0.99
5 40
Figure USRE041614-20100831-C00122
 		1.9 0.86 0.34 1.9
6 39
Figure USRE041614-20100831-C00123
 		4.0 0.91 0.32 1.1
7 43
Figure USRE041614-20100831-C00124
 		4.5 2.1 0.94 2.6
8 45
Figure USRE041614-20100831-C00125
 		5.2 2.1 0.98 3.1
9 48
Figure USRE041614-20100831-C00126
 		30 6.0 2.5 7.7
10 37
Figure USRE041614-20100831-C00127
 		49 19 11 26
11 38
Figure USRE041614-20100831-C00128
 		56 23 15 28
12 41
Figure USRE041614-20100831-C00129
 		2.5 1.1 0.62 1.5
13 42
Figure USRE041614-20100831-C00130
 		2.9 1.9 0.73 2.0
TABLE 7
Activity Data for Additional Analogs
Figure USRE041614-20100831-C00131
Mela-
No. Analog (X1 Group) Lung Colon noma Prostate
59
Figure USRE041614-20100831-C00132
 		0.81 0.40 0.23 0.49
69
Figure USRE041614-20100831-C00133
 		9.6 2.0 0.72 2.1
62
Figure USRE041614-20100831-C00134
 		1.7 0.65 0.33 0.82
63
Figure USRE041614-20100831-C00135
 		2.0 0.37 0.22 0.40
70
Figure USRE041614-20100831-C00136
 		66 19 8.9 25
Figure USRE041614-20100831-C00137
66
Figure USRE041614-20100831-C00138
 		8.6 1.3 0.92 2.1
TABLE 8
Additional Biological Data
Cancer IC50
Structure Cells (ng/mL)
Figure USRE041614-20100831-C00139
 		Lung Colon Melan. Prost. 1.1 0.53 0.30 0.45
Figure USRE041614-20100831-C00140
 		Lung Colon Melan. Prost. 160 51 46 68
Figure USRE041614-20100831-C00141
 		Lung Colon Melan. Prost. 21 3.5 1.9 3.3
Figure USRE041614-20100831-C00142
 		Lung Colon Melan. Prost. 40 20 8.3 17
Figure USRE041614-20100831-C00143
 		Lung Colon Melan. Prost. 4.7 1.3 0.47 2.0
Figure USRE041614-20100831-C00144
 		Lung Colon Melan. Prost. >100 >100 >100 >100
Figure USRE041614-20100831-C00145
 		Lung Colon Melan. Prost. 48 13 2.1 9.0
Figure USRE041614-20100831-C00146
 		Lung Colon Melan. Prost. 0.67 0.39 0.13 0.34
Figure USRE041614-20100831-C00147
 		Lung Colon Melan. Prost. 1.9 0.70 0.24 0.73
Figure USRE041614-20100831-C00148
 		Lung Colon Melan. Prost. >100 24 9.0 22
Based upon the tests performed to date, it is believed that the compounds of the present invention will serve as useful antitumor agents in mammals, particularly in humans.
Antitumor compounds are typically administered in unit dosage form. Each unit dose, as it pertains to the present invention, refers to a physically discrete unit suitable as unitary dosages for animals, each unit containing a predetermined quantity of active material calculated to produce the desired antitumor effect in association with the required diluent; i.e., carrier, or vehicle. The specifications for the novel unit dose of this invention are dictated by and are directly dependent on (a) the unique characteristics of the active material and the particular antitumor effect to be achieved, and (b) the limitations inherent in the art of compounding such active material for such use in mammals, particularly humans, as disclosed in detail herein, these being features of the present invention.
Unit dosage forms are typically prepared from the active compound by dispersement thereof in a physiologically tolerable (or acceptable) diluent or vehicle such as water, saline or phosphate-buffered saline, to form an aqueous composition. If necessary, other pharmaceutically acceptable solvents may be used. Such diluents are well known in the art and are discussed, for example, in Remington's Pharmaceutical Sciences, 16th Ed., Mack Publishing Company, Easton, Pa. (1980) at pages 1465-1467.
Dosage forms can also include an adjuvant as part of the diluent. Adjuvants such as complete Freund's adjuvant (CFA), incomplete Fruend's adjuvant (IFA) and alum are materials well known in the art, and are available commercially from several sources.
The quantity of active compound to be administered depends, inter alia, on the animal species to be treated, the subject animal's size, the size of the tumor being treated (if known), and the capacity of the subject to utilize the active compound. Precise amounts of the active compound required to be administered depend on the judgment of the practitioner and are peculiar to each individual, particularly where humans are the treated animals. Dosage ranges, however, can be characterized by a therapeutically effective blood concentration and can range from a concentration of the active compound of the present invention from about 0.01 μM to about 100 μM, preferably about 0.1 μM to 10 μM.
Suitable regimes for initial administration and booster injections are also variable, but are typified by an initial administration followed by repeated doses at one or more hour intervals by a subsequent injection or other administration. Alternatively, continuous intravenous infusion sufficient to maintain therapeutically effective concentrations in the blood are contemplated.
The present invention has been described in detail, including the preferred embodiments thereof. However, it will be appreciated that those skilled in the art, upon consideration of the present disclosure, may make modifications and/or improvements on this invention and still be within the scope and spirit of this invention as set forth in the following claims.

Claims (51)

1. Compounds having the formula:
Figure USRE041614-20100831-C00149
wherein each of the groups X1, X2, R1, R2, R3, R4, R5, R6, R7, R8 and R9 is independently selected from the group consisting of H, OH, OR′, SR′, SOR′, SO2R′, NO2, NH2, NHR′, N(R′)2, NHC(O)R′ NHC(═O)R′, CN, halogen, ═O, C(═O)H, C(═O)R′, CO2H, CO2R′, C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl, and substituted or unsubstituted heteroaromatic;
wherein each of the R′ groups is independently selected from the group consisting of H, OH, NH2, NO2, SH, CN, halogen, ═O, C(═O)H, C═O)CH3 C(═O)CH 3, CO2H, CO2CH3, C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, aryl, aralkyl, and heteroaromatic;
wherein each dotted circle represents one, two or three optional double bonds; and
wherein R7 and R8 may be are joined into a carbocyclic or heterocyclic ring system.
2. The compounds of claim 1 wherein X1 and X2 are independently selected from the group consisting of:
Figure USRE041614-20100831-C00150
or the formula:
Figure USRE041614-20100831-C00151
wherein Z is selected from the group consisting of:
Figure USRE041614-20100831-C00152
where n=1, 2, 3 . . . 20
wherein each R group is independently selected from the group consisting of H, OH, SH, NH2, NO2, halogen, nitro, C1-C6-alkyl, C1-C6-alkoxy, C1-C6-acyl, substituted or unsubstituted phenyl or substituted or unsubstituted benzyl.
3. Compounds of the formula:
Figure USRE041614-20100831-C00153
wherein each of the groups X1, X2, R1, R2, R3, R4, R5, R6, and R9 is independently selected from the group consisting of H, OH, OR′, SH, SR′, SOR′, SO2R′, NO2, NH2, NHR′, N(R′)2, NHC(O)R′ NHC(═O)R′, CN, halogen, ═O, C1-C6 alkyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl, and substituted or unsubstituted heteroaromatic;
wherein each of the R′ groups is independently selected from the group consisting of H, OH, NO2, NH2, SH, CN, halogen, ═O, C(═O)H, C(═O)CH3, CO2H, CO2CH3, C1-C6 alkyl, phenyl, benzyl, and heteroaromatic; and
wherein each dotted circle represents one, two or three optional double bonds.
4. The compounds of claim 3, wherein X1 and X2 are each independently selected from the group consisting of:
Figure USRE041614-20100831-C00154
or the formula:
Figure USRE041614-20100831-C00155
wherein Z is selected from the group consisting of:
Figure USRE041614-20100831-C00156
wherein each R group is independently selected from the group consisting of H, OH, SH, NH2, NO2, halogen, C1-C6-alkyl, C1-C6-alkoxy, C1-C6-acyl, aryl or alky-laryl alkylaryl.
5. The compound of the formula:
Figure USRE041614-20100831-C00157
and pharmaceutically acceptable salts and derivatives thereof, wherein:
R1 is H, OH, SH or NH2;
R2 is H, OH, OCH3;
R3 is H, OH, SH, NH2 or CH3;
R4 is H, OH, SH, NH2, OCH3, or halogen;
R5 is H or C1-C6 alkyl;
R6 is CN, OH, SH, NH2, OR, SR, or O(C═O)R;
R9 is C1-C6 alkyl;
wherein X1 is selected from the group consisting of:
Figure USRE041614-20100831-C00158
Figure USRE041614-20100831-C00159
Figure USRE041614-20100831-C00160
where each R group, which may be the same or be different, is selected from the group consisting of H, OH, SH, NH2, NO2, halogen, C1-C6-alkyl, C1-C6-alkoxy, C1-C6-acyl, alyl aryl or alkylaryl.
6. The compound of claim 5, wherein X1 is:
Figure USRE041614-20100831-C00161
7. The compound of claim 5, wherein X1 is:
Figure USRE041614-20100831-C00162
8. The compound of claim 5, wherein X1 is:
Figure USRE041614-20100831-C00163
9. The compound of claim 5, wherein X1 is:
Figure USRE041614-20100831-C00164
10. The compound of claim 5, wherein X1 is:
Figure USRE041614-20100831-C00165
11. The compound of claim 5, wherein X1 is:
Figure USRE041614-20100831-C00166
12. The compound of claim 5, wherein X1 is:
Figure USRE041614-20100831-C00167
13. The compound of claim 9 5, wherein X1 is:
Figure USRE041614-20100831-C00168
14. The compound of claim 5, wherein X1 is:
Figure USRE041614-20100831-C00169
15. The compound of claim 5, wherein X1 is:
Figure USRE041614-20100831-C00170
16. The compound of claim 5, wherein X1 is:
Figure USRE041614-20100831-C00171
17. The compound of claim 5, wherein X1 is:
Figure USRE041614-20100831-C00172
18. The compound of claim 5, wherein X1 is:
Figure USRE041614-20100831-C00173
19. Pharmaceutical compositions comprising an effective antitumor amount of a compound of the formula:
Figure USRE041614-20100831-C00174
and pharmaceutically acceptable salts and derivatives thereof, wherein:
R1 is H, OH, SH or NH2;
R2 is H, OH, OCH3;
R3 is H, OH, SH, NH2 or CH3;
R4 is H, OH, SH, NH2, OCH3, or halogen;
R5 is H or C1-C6 alkyl;
R6 is CN, OH, SH, NH2, OR, SR, or O(CO)R;
R9 is C1-C6 alkyl;
wherein X1 is selected from the group consisting of:
Figure USRE041614-20100831-C00175
Figure USRE041614-20100831-C00176
and
wherein each R group, which many be the same or be different, is selected from the group consisting of H, OH, SH, NH2, NO2, halogen, C1-C6-alkyl, C1-C6-alkoxy, C1-C6-acyl, aryl or alkylaryl.
20. The pharmaceutical composition of claim 19, wherein X1 is:
Figure USRE041614-20100831-C00177
21. The pharmaceutical composition of claim 19, wherein X1 is:
Figure USRE041614-20100831-C00178
22. The pharmaceutical composition of claim 19, wherein X1 is:
Figure USRE041614-20100831-C00179
23. The pharmaceutical composition of claim 19, wherein X1 is:
Figure USRE041614-20100831-C00180
24. The pharmaceutical composition of claim 19, wherein X1 is:
Figure USRE041614-20100831-C00181
25. The pharmaceutical composition of claim 19, wherein X1 is:
Figure USRE041614-20100831-C00182
26. The pharmaceutical composition of claim 19, wherein X1 is:
Figure USRE041614-20100831-C00183
27. The pharmaceutical composition of claim 19, wherein X1 is:
Figure USRE041614-20100831-C00184
28. The pharmaceutical composition of claim 19, wherein X1 is:
Figure USRE041614-20100831-C00185
29. The pharmaceutical composition of claim 19, wherein X1 is:
Figure USRE041614-20100831-C00186
30. The pharmaceutical composition of claim 19, wherein X1 is:
Figure USRE041614-20100831-C00187
31. The pharmaceutical composition of claim 19, wherein X1 is:
Figure USRE041614-20100831-C00188
32. The pharmaceutical composition of claim 19, wherein X1 is:
Figure USRE041614-20100831-C00189
33. A method of treating tumors selected from the group consisting of lung cancer, colon cancer and prostate cancer in mammals comprising administering to a mammal in need of such treatment, an effective antitumor amount of a compound of the formula:
Figure USRE041614-20100831-C00190
and pharmaceutically acceptable salts and derivatives thereof, wherein:
R1 is H, OH, SH or NH2;
R2 is H, OH, OCH3;
R3 is H, OH, SH, NH2 or CH3;
R4 is H, OH, SH, NH2, OCH3, or halogen;
R5 is H or C1-C6 alkyl;
R6 is CN, OH, SH, NH2, OR, SR, or O(CO)R;
R9 is C1-C6 alkyl,
wherein X1 is selected from the group consisting of:
Figure USRE041614-20100831-C00191
Figure USRE041614-20100831-C00192
and
wherein each R group, which may be the same or be different, is selected from the group consisting of H, OH, SH, NH2, NO2, halogen, C1-C6-alkyl, C1-C6-alkoxy, C1-C6-acyl, aryl or alkylaryl.
34. The method of treatment of claim 33, wherein X1 is:
Figure USRE041614-20100831-C00193
35. The method of treatment of claim 33, wherein X1 is:
Figure USRE041614-20100831-C00194
36. The method of treatment of claim 33, wherein X1 is:
Figure USRE041614-20100831-C00195
37. The method of treatment of claim 33, wherein X1 is:
Figure USRE041614-20100831-C00196
38. The method of treatment of claim 33, wherein X1 is:
Figure USRE041614-20100831-C00197
39. The method of treatment of claim 33, wherein X1 is:
Figure USRE041614-20100831-C00198
40. The method of treatment of claim 33, wherein X1 is:
Figure USRE041614-20100831-C00199
41. The method of treatment of claim 33, wherein X1 is:
Figure USRE041614-20100831-C00200
42. The method of treatment of claim 33, wherein X1 is:
Figure USRE041614-20100831-C00201
43. The method of treatment of claim 33, wherein X1 is:
Figure USRE041614-20100831-C00202
44. The method of treatment of claim 33, wherein X1 is:
Figure USRE041614-20100831-C00203
45. The method of treatment of claim 33, wherein X1 is:
Figure USRE041614-20100831-C00204
46. The method of treatment of claim 33, wherein X1 is:
Figure USRE041614-20100831-C00205
47. The compound of the formula:
Figure USRE041614-20100831-C00206
and pharmaceutically acceptable salts and derivatives thereof, wherein:
R 1 is H, OH, SH or NH 2 ;
R 2 is H, OH, OCH 3 ;
R 3 is H, OH, SH, NH 2 or CH 3 ;
R 4 is H, OH, SH, NH 2 , OCH 3 , or halogen;
R 5 is H or C 1 -C 6 alkyl;
R 6 is CN, OH, SH, NH 2 , OR, SR, or O(C═O)R;
R 9 is C 1 -C 6 alkyl;
wherein X 1 is selected from the group consisting of:
Figure USRE041614-20100831-C00207
where each R group, which may be the same or be different, is selected from the group consisting of H, OH, SH, NH 2 , NO 2 , halogen, C 1 -C 6-alkyl, C 1 -C 6-alkoxy, C 1 -C 6-acyl, aryl or alkylaryl.
48. Pharmaceutical compositions comprising an effective antitumor amount of a compound of the formula:
Figure USRE041614-20100831-C00208
and pharmaceutically acceptable salts and derivatives thereof, wherein:
R 1 is H, OH, SH or NH 2 ;
R 2 is H, OH, OCH 3 ;
R 3 is H, OH, SH, NH 2 or CH 3 ;
R 4 is H, OH, SH, NH 2 , OCH 3 , or halogen;
R 5 is H or C 1 -C 6 alkyl;
R 6 is CN, OH, SH, NH 2 , OR, SR, or O(C═O)R;
R 9 is C 1 -C 6 alkyl;
wherein X 1 is selected from the group consisting of:
Figure USRE041614-20100831-C00209
and
wherein each R group, which many be the same or be different, is selected from the group consisting of H, OH, SH, NH 2 , NO 2 , halogen, C 1 -C 6-alkyl, C 1 -C 6-alkoxy, C 1 -C 6-acyl, aryl or alkylaryl.
49. A method of treating tumors selected from the group consisting of lung cancer, colon cancer and prostate cancer in mammals comprising administering to a mammal in need of such treatment, an effective antitumor amount of a compound of the formula:
Figure USRE041614-20100831-C00210
and pharmaceutically acceptable salts and derivatives thereof, wherein:
R 1 is H, OH, SH or NH 2 ;
R 2 is H, OH, OCH 3 ;
R 3 is H, OH, SH, NH 2 or CH 3 ;
R 4 is H, OH, SH, NH 2 , OCH 3 , or halogen;
R 5 is H or C 1 -C 6 alkyl;
R 6 is CN, OH, SH, NH 2 , OR, SR, or O(C═O)R;
R 9 is C 1 -C 6 alkyl;
wherein X 1 is selected from the group consisting of:
Figure USRE041614-20100831-C00211
and
wherein each R group, which may be the same or be different, is selected from the group consisting of H, OH, SH, NH 2 , NO 2 , halogen, C 1 -C 6-alkyl, C 1 -C 6-alkoxy, C 1 -C 6-acyl, aryl or alkylaryl.
50. Pharmaceutical compositions comprising an effective antitumor amount of a compound of the formula:
Figure USRE041614-20100831-C00212
and pharmaceutically acceptable salts and derivatives thereof, wherein:
X 1 is selected from the group consisting of:
Figure USRE041614-20100831-C00213
and
Figure USRE041614-20100831-C00214
wherein each R group, which many be the same or be different, is selected from the group consisting of H, OH, SH, NH 2 , NO 2 , halogen, C 1 -C 6-alkyl, C 1 -C 6-alkoxy, C 1 -C 6-acyl, aryl or alkylaryl.
51. A method of treating tumors selected from the group consisting of lung cancer, colon cancer and prostate cancer in mammals comprising administering to a mammal in need of such treatment, an effective antitumor amount of a compound of the formula:
Figure USRE041614-20100831-C00215
and pharmaceutically acceptable salts and derivatives thereof, wherein:
X 1 is selected from the group consisting of:
Figure USRE041614-20100831-C00216
and
wherein each R group, which may be the same or be different, is selected from the group consisting of H, OH, SH, NH 2 , NO 2 , halogen, C 1 -C 6-alkyl, C 1 -C 6-alkoxy, C 1 -C 6-acyl, aryl or alkylaryl.
US10/738,973 1998-09-30 2003-12-17 Synthetic analogs of ecteinascidin-743 Expired - Lifetime USRE41614E1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US10/738,973 USRE41614E1 (en) 1998-09-30 2003-12-17 Synthetic analogs of ecteinascidin-743

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US09/165,892 US6124292A (en) 1998-09-30 1998-09-30 Synthetic analogs of ecteinascidin-743
US09/510,315 US6348467B1 (en) 1998-09-30 2000-02-22 Synthetic analogs of ecteinascidin-743
US10/738,973 USRE41614E1 (en) 1998-09-30 2003-12-17 Synthetic analogs of ecteinascidin-743

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
US09/510,315 Reissue US6348467B1 (en) 1998-09-30 2000-02-22 Synthetic analogs of ecteinascidin-743

Publications (1)

Publication Number Publication Date
USRE41614E1 true USRE41614E1 (en) 2010-08-31

Family

ID=22600906

Family Applications (3)

Application Number Title Priority Date Filing Date
US09/165,892 Expired - Lifetime US6124292A (en) 1998-09-30 1998-09-30 Synthetic analogs of ecteinascidin-743
US09/510,315 Ceased US6348467B1 (en) 1998-09-30 2000-02-22 Synthetic analogs of ecteinascidin-743
US10/738,973 Expired - Lifetime USRE41614E1 (en) 1998-09-30 2003-12-17 Synthetic analogs of ecteinascidin-743

Family Applications Before (2)

Application Number Title Priority Date Filing Date
US09/165,892 Expired - Lifetime US6124292A (en) 1998-09-30 1998-09-30 Synthetic analogs of ecteinascidin-743
US09/510,315 Ceased US6348467B1 (en) 1998-09-30 2000-02-22 Synthetic analogs of ecteinascidin-743

Country Status (13)

Country Link
US (3) US6124292A (en)
EP (1) EP1117297B1 (en)
JP (1) JP4638985B2 (en)
AT (1) ATE303063T1 (en)
AU (1) AU765439B2 (en)
CA (1) CA2345297C (en)
DE (1) DE69927039T2 (en)
DK (1) DK1117297T3 (en)
ES (1) ES2249028T3 (en)
HK (1) HK1039727B (en)
MX (1) MXPA01003339A (en)
NZ (1) NZ510734A (en)
WO (1) WO2000018233A1 (en)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100197695A1 (en) * 2001-07-17 2010-08-05 Valentin Martinez Antitumoral derivatives of et-743
US20110070232A1 (en) * 2008-05-16 2011-03-24 Pharma Mar, S.A. Combination Therapy with an Antitumor Alkaloid
US20110076343A1 (en) * 2008-05-16 2011-03-31 Pharma Mar, S.A. Multiple Myeloma Treatments
US9428524B2 (en) 2010-05-25 2016-08-30 Pharma Mar, S.A. Synthetic process for the manufacture of ecteinascidin compounds

Families Citing this family (28)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6316214B1 (en) * 1998-05-11 2001-11-13 The Board Of Trustees Of The University Of Illinois ETM-775 metabolite of ecteinascidin 743
US6124292A (en) * 1998-09-30 2000-09-26 President And Fellows Of Harvard College Synthetic analogs of ecteinascidin-743
MY164077A (en) * 1999-05-13 2017-11-30 Pharma Mar Sa Compositions and uses of et743 for treating cancer
GB9918178D0 (en) * 2000-05-15 1999-10-06 Pharma Mar Sa Synthetic methods
AR035842A1 (en) * 1999-05-14 2004-07-21 Pharma Mar Sa METHOD OF HEMISINTESIS FOR THE FORMATION OF INTERMEDIARY AND DERIVATIVE COMPOUNDS AND OF STRUCTURES RELATED TO ECTEINASCIDINE AND TETRAHYDROISOCHINOLINPHENOLS AND INTERMEDIARY APPLICATION COMPOUNDS IN SUCH METHOD
JP2003520801A (en) * 2000-01-19 2003-07-08 ザ・トラスティーズ・オブ・コランビア・ユニバーシティー・イン・ザ・シティー・オブ・ニューヨーク Saframycin-ectineacidin family of compounds, their use and synthesis
EP1255759B1 (en) * 2000-02-11 2007-08-01 President And Fellows of Harvard College Synthetic process for an intermediate for ecteinascidin and phthalascidin compounds
US6815544B2 (en) 2000-02-11 2004-11-09 President And Fellows Of Harvard College Synthetic process for an intermediate for ecteinascidin and phthalascidin compounds
ATE299146T1 (en) * 2000-04-12 2005-07-15 Pharma Mar Sa ECTEINASCHIDINE DERIVATIVES WITH ANTI-CANCER EFFECT
US7919493B2 (en) * 2000-04-12 2011-04-05 Pharma Mar, S.A. Anititumoral ecteinascidin derivatives
MXPA02011319A (en) * 2000-05-15 2003-06-06 Pharma Mar Sa Antitumoral analogs of et 743.
US7420051B2 (en) * 2000-05-15 2008-09-02 Pharma Mar, S.A. Synthetic process for the manufacture of an ecteinaschidin compound
AU3956502A (en) * 2000-11-03 2002-05-27 Harvard College Saframycins, analogues and uses thereof
JP4391083B2 (en) * 2000-11-06 2009-12-24 ファルマ・マール・ソシエダード・アノニマ Effective anti-tumor treatment
GB0119243D0 (en) * 2001-08-07 2001-10-03 Pharma Mar Sa Antitumoral analogs of ET-743
GB0202544D0 (en) * 2002-02-04 2002-03-20 Pharma Mar Sa The synthesis of naturally occuring ecteinascidins and related compounds
US7183054B2 (en) * 2003-06-03 2007-02-27 President And Fellows Of Harvard College Assay for identifying biological targets of polynucleotide-binding compounds
UA87877C2 (en) * 2004-10-26 2009-08-25 Фарма Мар С.А., Сосьедад Униперсональ Pegylated liposomal doxorubicin in combination with ecteinescidin 743
KR101188691B1 (en) * 2004-10-29 2012-10-09 파르마 마르 에스.에이. Formulations comprising ecteinascidin and a disaccharide
GB0522082D0 (en) * 2005-10-31 2005-12-07 Pharma Mar Sa Formulations
WO2011048210A1 (en) 2009-10-22 2011-04-28 Pharma Mar, S.A. PDGFR-α AS RESPONSE MARKER FOR PM00104 TREATMENT
CA2995033C (en) 2010-11-12 2020-04-28 Pharma Mar, S.A. Combination therapy with an antitumor alkaloid
CN107522698B (en) * 2016-06-20 2021-12-28 浙江海正药业股份有限公司 Preparation method and intermediate of trabectedin
TWI807411B (en) 2017-04-27 2023-07-01 西班牙商瑪製藥股份有限公司 Antitumoral compounds
US10597378B2 (en) 2017-09-08 2020-03-24 National Health Research Institutes Tetrahydroisoquinolines for use as MOR/NOP dual agonists
CN111518110B (en) * 2019-02-01 2023-11-03 博瑞生物医药（苏州）股份有限公司 Preparation method of ecteinascidin compound and intermediate thereof
EP4061372A1 (en) 2019-11-21 2022-09-28 Pharma Mar, S.A. Methods of treating small cell lung cancer with lurbinectedin formulations
WO2024186263A1 (en) * 2023-03-07 2024-09-12 Axcynsis Therapeutics Pte. Ltd. Antibody-drug conjugates comprising trabectedin and lurbinectedin derivatives

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4248863A (en) 1977-09-12 1981-02-03 Tadashi Arai Antibiotics saframycins A, B, C, D and E and process for producing the same
US4372947A (en) 1980-03-26 1983-02-08 Tadashi Arai Antibiotic saframycin S and process for producing the same
JPS632991A (en) * 1986-06-23 1988-01-07 Akinori Kubo Novel isoquinoline derivative
US5089273A (en) 1986-06-09 1992-02-18 Board Of Trustees Of The University Of Illinois Ecteinascidins 729, 743, 745, 759A, 759B and 770
US5256663A (en) 1986-06-09 1993-10-26 The Board Of Trustees Of The University Of Illinois Compositions comprising ecteinascidins and a method of treating herpes simplex virus infections therewith
US5478932A (en) 1993-12-02 1995-12-26 The Board Of Trustees Of The University Of Illinois Ecteinascidins
US5721362A (en) 1996-09-18 1998-02-24 President And Fellows Of Harvard College Process for producing ecteinascidin compounds
US6124292A (en) * 1998-09-30 2000-09-26 President And Fellows Of Harvard College Synthetic analogs of ecteinascidin-743

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5942382A (en) * 1982-09-02 1984-03-08 Yoshitomi Pharmaceut Ind Ltd Safracin derivative
JPS6158593A (en) * 1984-08-30 1986-03-25 Tadashi Arai Novel saframycin a derivtive and its preparation
EP0233841A1 (en) * 1986-02-18 1987-08-26 Arai, Tadashi, Prof. Quinone derivatives and process for their preparation
EP0262085A1 (en) * 1986-08-15 1988-03-30 Gesellschaft für Biotechnologische Forschung mbH (GBF) Antibiotics from myxococcus
EP0329606A3 (en) * 1988-02-12 1991-04-03 Ciba-Geigy Ag Antibiotics from myxocuccos

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4248863A (en) 1977-09-12 1981-02-03 Tadashi Arai Antibiotics saframycins A, B, C, D and E and process for producing the same
US4372947A (en) 1980-03-26 1983-02-08 Tadashi Arai Antibiotic saframycin S and process for producing the same
US5089273A (en) 1986-06-09 1992-02-18 Board Of Trustees Of The University Of Illinois Ecteinascidins 729, 743, 745, 759A, 759B and 770
US5256663A (en) 1986-06-09 1993-10-26 The Board Of Trustees Of The University Of Illinois Compositions comprising ecteinascidins and a method of treating herpes simplex virus infections therewith
JPS632991A (en) * 1986-06-23 1988-01-07 Akinori Kubo Novel isoquinoline derivative
US5478932A (en) 1993-12-02 1995-12-26 The Board Of Trustees Of The University Of Illinois Ecteinascidins
US5654426A (en) 1993-12-02 1997-08-05 Univ Illinois Ecteinascidins
US5721362A (en) 1996-09-18 1998-02-24 President And Fellows Of Harvard College Process for producing ecteinascidin compounds
US6124292A (en) * 1998-09-30 2000-09-26 President And Fellows Of Harvard College Synthetic analogs of ecteinascidin-743

Non-Patent Citations (39)

* Cited by examiner, † Cited by third party
Title
"Chemical Prospectors Scour The Seas for Promising Drugs", Science, 1994, vol. 266, pp. 1324-1325.
Akinori Kubo et al., Stereoselective Total Synthesis of (±)-Saframycin B, J. Org. Chem., 53, pp. 4295-4310, 1988.
Amy Wright et al., "Antitumor Tetrahydroisoquinoline Alkaloids from the Colonial Ascidian Ecteinasccidia turbinata", J. Org. Chem. 1990, 55, pp. 4508-4512.
Corey et al., "Enantioselective Total Synthesis of Ecteinascidin 743", J. Am. Chem. Soc., vol. 118, No. 38, pp. 9201-9203, Sep. 25, 1996.
E. J. Corey et al., "Total Synthesis of a Potent Thromboxane A2 Antagonsit", Tetrahedron Letters, vol. 31, No. 27, pp. 3833-3836 (1990).
E.H.R. Barton, et al., "Synthesis and Properties of a Series of Sterically Hindered Guanidine Bases", J.Chem.Soc. 1982, pp. 2085-2090.
E.J. Corey, et al. "Enantioselective Total Synthesis of Ecteinascidin", J.Am.Chem.Soc. 1996, vol. 118, pp. 9202-9203.
E.V. Ganin et al., "Reaction of Aromatic Di- and Tetracarboxylic Anhydrides with Amides", J. Org. Chem. USSR 1987, vol. 23, pp. 981-983.
J. W. Lown et al., "Molecular Mechanisms of Binding and Single-Strand Scission of Deoxyribonucleic Acid by the Antitumor Antibiotics Saframycins A and C", Biochemistry, vol. 21, No. 3, pp. 419-428 (1982).
K. Koenig, "Asymmetric Synthesis", vol. 5, 1985, Academic Press, Inc., Orlando, FL., pp. 73-101.
Kobo Akiyoshi et al. "Studies on the total syn . . . Saframycin B"; Meiji Coll. Phram. 28th, 465-72 (1986). *
Kobo-Akiyoshi et al. "Studies on the total synthesis . . . Saframycin B"; Meiji Coll. Pharm. 28th, 465-72, Jan. 1986. *
L. Campayo et al., "Diazapolycyclic Compounds: XXV. Improved Synthesis of 6-Substituted 2,3-Dihydrobenzo [g]phtalazine-1,4-dione Derivatives", Synthesis Communications, 1985, pp. 197-200.
M. Alexiou et al., "Nucleophilic Displacement of the Nitro Group In 2- and 4-Nitronaphthalic-1,8-Anhydrides and Their Derivatives", Tetrahedron Letters, vol. 22, No. 24, pp. 2303-2306 (1981).
M. Nakagawa et al., "Total Synthesis of (-)-Eudistomin L and (-)-Debromoeudistomin L", J. Am. Chem. Soc. 1989, vol. 111, pp. 2722-2724.
M. Nakagawa et al., "Total Synthesis of (−)-Eudistomin L and (−)-Debromoeudistomin L", J. Am. Chem. Soc. 1989, vol. 111, pp. 2722-2724.
M. Newman et al., "A Convenient Synthesis of 1,2-Naphthalic Anhydride", J. Org. Chem., vol. 41, No. 24, pp. 3925 (1976).
M. Zmijewski et al., "The In Vitro Interaction of Naphthyridinomycin with Deoxyribonucleic Acids", Chem.-Biol. Interactions, vol. 52, pp. 361-375 (1985).
N. Saito et al., "Synthesis of Saframycins. 3. Preparation of a Key Tricyclic Lactam Intermediate to Saframycin A" J Org Chem vol. 54 No. 22 (1989) pp. 5392-5395.
Naoki Saito et al., Synthesis of Saframycins. VII. The Synthesis of Novel Ranieramycin Congeners, Heterocycles, vol. 32, No. 6, pp. 1203-1214, 1991.
Plakidin et al., "Reaction of Naphthalic Anhydride and Its Substituted Derivatives with Acid Amides", J. Org. Chem. USSR 1982, vol. 18, pp. 1754-1755.
R. Sakai et al., "Additional and Antitumor Ecteinascidins from a Caribbean Tunicate: Crystal Structures and Activities in vivo", Proc. Natl. Acad. Sci. USA, vol. 89, pp. 11456-11460.
R. Sakai et al., "Ecteinascidins: Putative Biosynthetic Precursors and Absolute Stereochemistry", J. Am. Chem. Soc. 1996, vol. 118, pp. 9017-9023.
Rinehart et al., "Bioactive Compounds from Aquatic and Terrestrial Sources", Journal of Natural Products, vol. 53, No. 4, pp. 771-792 (1990).
Rinehart et al., "Biologically Active Natural Products", Pure & App. Chem., vol. 62, No. 7, pp. 1277-1280 (1990).
Rinehart et al., "Ecteinascidins 729, 743, 745, 759A. 759Bm and 770: Potent Antitumor Agents from the Caribbean Tunicate Ecteinascidia turbinata", J. Org. Chem. 1990, 55 pp. 4512-4515.
Sakai et al., Ecteinascidins: Putative Biosynthetic Precursors and Absolute Steriochemistry, J. Am. Chem. Soc., 1996, vol. 118, pp. 9017-9023.
T. Fukuyama et al., "Stereocontrolled Total Synthesis of Saframycin B", J. Am. Chem. Soc. 1982, vol. 104, pp. 4957-4958.
T. Fukuyama et al., "Total Synthesis of Saframycin B", J. Am. Chem. Soc. 1990, vol. 112, pp. 3712-3715.
T. Fukuyama et al., "Total Synthesis of Saframycin B", J. Am. Chem. Soc. 1990, vol. 112, pp. 3713-3715.
Tohru Fukuyama et al., Stereocontrolled Total Synthesis of (±)-Saframycin B, J. Am. Chem. Soc., 104, pp. 4957-4958, 1982.
Tohru Fukuyama et al., Total Synthesis of (±)-Saframycin A, J. Am. Chem. Soc., 112, pp. 3712-3713, 1990.
W. Clark Still et al., "Rapid Chromatographic Technique for Preparative Separations with Moderate Resolution", J. Org. Chem., vol. 43, No. 14, (1978), pp. 2923-2925.
W. Kofron et al., "A Convenient Method for Estimation of Alkyllithium Concentrations", J. Org. Chem., vol. 41, No. 10, (1976), pp. 1879-1880.
Weidner-Wells et al., "Photochemical Approach to the Synthesis of the Pyrrolo[1,4] benzodiazepine Antibiotics", J. Org. Chem. 1989, vol. 54, pp. 5746-5758.
Y. Guan et al., "Molecular and Crystal Structures of Ecteinascidins: Potent Antitumor Compounds from the Caribbean Tunicate Ecteinascidia turbinata", Journal of Biomolecular Structure & Dynamics, vol. 10, pp. 7930817 (1993).
Y. Ito, "High Speed Countercurrent Chromatography", CRC Critical Reviews in Analytical Chemistry, vol. 17, Issue 1, pp. 65-143.
Y. Pommier et al., "DNA Sequence-and Structure-Selective Alkylation of Guanine N2 in the DNA Minor Groove by Ecteinascidin 743, a Potent Antitumor Compound from the Caribbean Tunicate Ecteinascidia turbinata", Biochemistry 1996, vol. 35, pp. 13303-13309.
Y.Guan, et al., "Molecular and Crystal Structures of Ecteinascidins: Potent Antitumor Compounds from the Caribbean Tunicate Ecteinascidia turbinata", J. Biomolecular Structure & Dynamics, 1993, vol. 10, No. 5, pp. 793-818.

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100197695A1 (en) * 2001-07-17 2010-08-05 Valentin Martinez Antitumoral derivatives of et-743
US8076337B2 (en) 2001-07-17 2011-12-13 Pharma Mar, S.A. Antitumoral derivatives of ET-743
US20110070232A1 (en) * 2008-05-16 2011-03-24 Pharma Mar, S.A. Combination Therapy with an Antitumor Alkaloid
US20110076343A1 (en) * 2008-05-16 2011-03-31 Pharma Mar, S.A. Multiple Myeloma Treatments
US8435992B2 (en) 2008-05-16 2013-05-07 Pharma Mar S.A. Multiple myeloma treatments
US9428524B2 (en) 2010-05-25 2016-08-30 Pharma Mar, S.A. Synthetic process for the manufacture of ecteinascidin compounds

Also Published As

Publication number Publication date
HK1039727A1 (en) 2002-05-10
NZ510734A (en) 2003-10-31
EP1117297A4 (en) 2002-02-27
MXPA01003339A (en) 2002-07-02
CA2345297A1 (en) 2000-04-06
AU765439B2 (en) 2003-09-18
WO2000018233A1 (en) 2000-04-06
CA2345297C (en) 2009-12-22
EP1117297B1 (en) 2005-08-31
DK1117297T3 (en) 2006-01-09
ES2249028T3 (en) 2006-03-16
JP4638985B2 (en) 2011-02-23
US6124292A (en) 2000-09-26
US6348467B1 (en) 2002-02-19
EP1117297A1 (en) 2001-07-25
DE69927039D1 (en) 2005-10-06
ATE303063T1 (en) 2005-09-15
JP2002525296A (en) 2002-08-13
DE69927039T2 (en) 2006-06-22
AU6165099A (en) 2000-04-17
HK1039727B (en) 2006-01-06

Similar Documents

Publication Publication Date Title
USRE41614E1 (en) Synthetic analogs of ecteinascidin-743
CN102149717B (en) Dioxa-bicyclo[3.2.1]octane-2,3,4-triol derivatives
KR100886496B1 (en) Antitumor Actinidine Derivatives
US8012975B2 (en) Antitumoral analogs of ET-743
US8076337B2 (en) Antitumoral derivatives of ET-743
US6936714B2 (en) Compounds of the saframycin-ecteinascidin series, uses, and synthesis thereof
JPH08337584A (en) Fused hexacyclic amino compound, pharmaceutical containing the same, and process for producing the same
US6569859B1 (en) Synthetic analogs of ecteinascidin-743
US20080033001A1 (en) Derivatives of Variolin B
US8846734B2 (en) Diazonamide analogs
EP0055602A2 (en) Oxazolidinedione sulfide compounds

Legal Events

Date Code Title Description
FEPP Fee payment procedure

Free format text: PAYOR NUMBER ASSIGNED (ORIGINAL EVENT CODE: ASPN); ENTITY STATUS OF PATENT OWNER: LARGE ENTITY

FPAY Fee payment

Year of fee payment: 12