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  • C07C311/22—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound oxygen atoms
  • C07C311/29—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound oxygen atoms having the sulfur atom of at least one of the sulfonamide groups bound to a carbon atom of a six-membered aromatic ring
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  • C07C2601/14—The ring being saturated

Definitions

• This invention relates to acetylenic aryl sulfonamide and phosphinic acid amide hydroxamic acids which act as inhibitors of TNF- ⁇ converting enzyme
• TNF- ⁇ converting enzyme catalyzes the formation of TNF- ⁇ from membrane bound TNF- ⁇ precursor protein.
• TNF- ⁇ is a pro-inflammatory cytokine that is believed to have a role in rheumatoid arthritis [Shire, M. G.; Muller, G. W. Exp. Opin. Ther. Patents 1998, 8(5), 531; Grossman, J. M.; Brahn, E. J. Women's Health 1997, 6(6), 627; Isomaki, P.; Punnonen, J. Ann. Med. 1997, 29, 499; Camussi, G.; Lupia, E. Drugs, 1998, 55(5), 613.] septic shock [Mathison, et. al.
• sulfonamide hydroxamic acid MMP/TACE inhibitors in which a 2 carbon chain separates the hydroxamic acid and the sulfonamide nitrogen, as shown below, are disclosed in WIPO international publications WO9816503, WO9816506, WO9816514 and WO9816520 and U. S. patent 5,776,961.
• Sulfonamide MMP/TACE inhibitors in which a thiol is the zinc chelating group, as shown below, have been disclosed in WIPO international application 9803166.
• Y the sulfonyl or phosphinyl aryl
• These compounds provide enhanced levels of inhibition of the activity of TACE in vitro and in a cellular assay and/or selectivty over MMP-1. These compounds may therefore be used in the treatment of diseases mediated by TNF.
• TACE and MMP inhibiting ortho-sulfonamido hydroxamic acids of the present invention are represented by the formula:
• Z is O, NH, CI ⁇ or S;
• R 5 is hydrogen or alkyl of 1-6 carbon atoms;
• R ⁇ and R 12 are, independently, hydrogen, alkyl of 1-6 carbon atoms, cycloalkyl of 3-6 carbon atoms, a 5-7 membered heteroaryl having 1-3 heteroatoms selected from N, NR 9 , S and O, a 5-7 membered heterocycloalkyl having 1 or 2 heteroatoms selected from N, NR,, S and O, or phenyl, and the optional double bond represented by the dotted line is present; or
• Preferred compounds of this invention include compounds of structure B wherein X is SO 2 .
• More preferred compounds of this invention include compounds of structure B wherein X is SO 2 and Y is a phenyl ring substituted at the 1- and 4-positions by X and Z, respectively.
• More preferred compounds of this invention include compounds of structure B wherein X is SO 2 N is a phenyl ring substituted at the 1- and 4-positions by X and Z, respectively, and Z is oxygen.
• More preferred compounds of this invention include compounds of structure B wherein X is SO 2 N is a phenyl ring substituted at the 1- and 4-positions by X and Z, respectively, Z is oxygen and R 6 and R, are hydrogen.
• More preferred compounds of this invention include compounds of structure B wherein X is SO 2 N is a phenyl ring substituted at the 1- and 4-positions by X and Z, respectively, Z is oxygen, R 6 andR, are hydrogen and R g is -CH 2 OH or methyl.
• Heteroaryl as used throughout, is a 5-10 membered mono- or bicyclic ring having from 1-3 heteroatoms selected from N, NR 9 , S and O. Heteroaryl is preferably
• Halogen means bromine, chlorine, fluorine, and iodine.
• Suitable substituents of aryl, heteroaryl, alkyl, alkenyl, alkynyl, and cycloalkyl include, but are not limited to halogen, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, cyclocalkyl of 3-6 carbon atoms, -OR ; ,, -CN, -CO-R-,, perfluoroalkyl of 1-4 carbon atoms, -O-perfluoroalkyl of 1-4 carbon atoms, -CONR..R 3 ,
• Suitable substituents of heterocycloalkyl groups of the present invention include, but are not limited to alkyl of 1-6 carbon atoms, cycloalkyl of 3-6 carbon atoms, phenyl, naphthyl, heteroaryl and heterocycloalkyl.
• salts can be formed from organic and inorganic acids, for example, acetic, propionic, lactic, citric, tartaric, succinic, fumaric, maleic, malonic, mandelic, malic, phthalic, hydrochloric, hydrobromic, phosphoric, nitric, sulfuric, methanesulfonic, naphthalenesulfonic, benzenesulfonic, toluenesulfonic, camphorsulfonic, and similarly known acceptable acids when a compound of this invention contains a basic moiety.
• Salts may also be formed from organic and inorganic bases, preferably alkali metal salts, for example, sodium, lithium, or potassium, when a compound of this invention contains an acidic moiety.
• the compounds of this invention are shown to inhibit the enzymes MMP-1, MMP-9, MMP- 13 and TNF- ⁇ converting enzyme (TACE) and are therefore useful in the treatment of arthritis, tumor metastasis, tissue ulceration, abnormal wound healing, periodontal disease, graft rejection, insulin resistance, bone disease and HIV infection.
• TACE TNF- ⁇ converting enzyme
• the compounds of the invention provide enhanced levels of inhibition of the activity of TACE in vitro and in cellular assay and/or enhanced selectivity over MMP-1 and are thus particularly useful in the treatment of diseases mediated by TNF.
• this invention provides a process for preparing compounds of formula 1 , as defined above, which comprises one of the following: a) reacting a compound of formula V:
• Removal of protecting groups as illustrated by process b) can be carried out by processes known in the art to provide the hydroxamic acid.
• a racemic mixture may be converted to a mixture of optically active diastereoisomers by reaction with a single enantiomer of a 'resolving agent' (for example by diastereomeric salt formation or formation of a covalent bond).
• the resulting mixture of optically active diastereoisomers may be separated by standard techniques (e.g. crystallisation or chromatography) and individual optically active diastereoisomers then treated to remove the 'resolving agent' thereby releasing the single enantiomer of the compound of the invention.
• Chiral chromatography using a chiral support, eluent or ion pairing agent
• the compounds of formula B may be isolated in the form of a salt of a pharmaceutically acceptable acid e.g. an organic or inorganic acid by treatment with an acid such as described above.
• a pharmaceutically acceptable acid e.g. an organic or inorganic acid by treatment with an acid such as described above.
• the resultant sulfonyl chloride, fluoride or bromide may be further converted into triazolide, imidazolide or benzothiazolide derivatives, where J is 1,2,4-triazolyl, imidazol-yl or benzotriazolyl, by reacting the compound with 1,2,4-triazole, imidazole or benzotriazole, respectively.
• R 6 , R. and R g are as defined above.
• Compounds 3, wherein R ⁇ is a t-butyl, benzyl, trialkylsilyl or other suitable masking group may then be deprotected by known methods to provide the hydroxamic acid 1.
• the acetylenic side chain may also be appended after sulfonylation or phosphorylation of the amino acid derivative, as shown in Scheme 5.
• the amino acid derivatives 4 and 8 can be sulfonylated or phosphorylated with compounds 20, where ZR 50 is hydroxy or protected hydroxy, thiol or amine, and, if necessary, alkylated with R,J as in Scheme 2, to give 21. Removal of the R 50 masking group to give 22 and subsequent alkylation of the resulting phenol, thiol or amine with 10 provides 7. In the case where ZR 50 is equal to OH, no deprotection step is required to give 22.
• the propargylic amine analogs of 7 can be synthesized as shown in Scheme 6 starting from the amino acid derivatives 4 and/or 8.
• Sulfonylation or phosphorylation with para-nitro aryl compound 23, for example 4-nitrobenzenesulfonyl chloride, followed by alkylation with R 5 J (for 4) using a base such as potassium carbonate or sodium hydride in DMF provides 24.
• Reduction of the nitro moiety with hydrogen and palladium on carbon, tin chloride or other known method to give aniline 25 and subsequent alkylation with 10 then provides 7.
• Aniline 25 may be derivatized with a suitable nitrogen protecting group, such as t-butoxycarbonyl, to give 26 prior to alkylation with 10 subsequent deprotection after the alkylation step.
• Scheme 6 :
• Acetylenic derivatives 7 are also accessible via the fluoro compounds 27, readily prepared from the amino acid derivatives 4 and/or 8 by reaction with fluoraryl 26, as shown in Scheme 7.
• the fluorine of 27 can also be displaced in a polar aprotic solvent with the propargylic derivative 29, where Z is O, S or NH, in the presence of a base such as sodium hydride, to give 7 directly.
• Scheme 7 :
• Example 8 (cis)-2-[Benzyl-(4-methoxy-benzenesuIfonyI)-amino]-cyclohexanecarboxyIic acid
• 0.240 g (0.522 mmol) of the product from Example 6 provided 0.207 g (98%) of the desired carboxylic acid as a white solid.
• Example 25 to give a white solid (0.825 g), mp 172-175°C. Analysis for
• the thiopeptide substrate is made up fresh as a 20 mM stock in 100% DMSO and the DTNB is dissolved in 100% DMSO as a 100 mM stock and stored in the dark at room temperature. Both the substrate and DTNB are diluted together to 1 mM with substrate buffer (50 mM HEPES pH 7.5, 5 mM CaCl2) before use. The stock of enzyme is diluted with buffer (50 mM HEPES, pH 7.5, 5 mM CaCl2, 0.02% Brij) to the desired final concentration.
• substrate buffer 50 mM HEPES pH 7.5, 5 mM CaCl2
• buffer 50 mM HEPES, pH 7.5, 5 mM CaCl2, 0.02% Brij
• a fluorescent peptide substrate is used.
• the peptide substrate contains a fluorescent group and a quenching group.
• the fluorescence that is generated is quantitated on the fluorescence plate reader.
• the assay is run in HCBC assay buffer (50mM
• HEPES pH 7.0, 5 mM Ca+ 2 , 0.02% Brij, 0.5% Cysteine
• the substrate is dissolved in methanol and stored frozen in 1 mM aliquots.
• substrate and enzymes are diluted in HCBC buffer to the desired concentrations.
• Compounds are added to the 96 well plate containing enzyme and the reaction is started by the addition of substrate.
• the reaction is read (excitation 340 nm, emission 444 nm) for 10 min. and the increase in fluorescence over time is plotted as a linear line.
• Compounds can be evaluated for their effect on basal (non-stimulated) shedding of the receptors by replacing the LPS with 50 ml/well of THP-1 media. Plates are placed into an incubator set at 5% CO2 and at 37° C. After 4 hours of incubation, 300 ml/well of tissue culture supernatant (TCS) is removed for use in an TNF- ⁇ ELISA. Following 24 hours of incubation, 700 ml/well of TCS is removed and used for analysis in TNF-R p75/80, TNF-R p55/60 and IL-8 ELISAs.
• TCS tissue culture supernatant
• the cells for each treatment group are collected by resuspension in 500 ⁇ l/well of THP-1 media and transferred into a FACS tube.
• Two ml/tube of a 0.5 mg/ml stock of propidium iodide (PI) (Boerhinger Mannheim cat. # 1348639) is added.
• the samples are run on a Becton Dickinson FaxCaliber FLOW cytometry machine and the amount of dye taken up by each cell is measured in the high red wavelength (FL3). Only cells with compromised membranes (dead or dying) can take up PI.
• the percent of live cells is calculated by the number of cells not stained with PI, divided by the total number of cells in the sample.
• IC 50 values for each compound are calculated by non-linear regression analysis using customized software utilizing the JUMP statistical package.
• Liquid carriers may be used in preparing solutions, suspensions, emulsions, syrups and elixirs.
• the active ingredient of this invention can be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture of both or pharmaceutically acceptable oils or fat.
• a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture of both or pharmaceutically acceptable oils or fat.
• the liquid carrier can contain other suitable pharmaceutical additives such a solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colors, viscosity regulators, stabilizers or osmo-regulators.

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• 2000
  • 2000-01-26 AR ARP000100325A patent/AR035478A1/es not_active Application Discontinuation
  • 2000-01-27 KR KR1020017009153A patent/KR20010089617A/ko not_active Application Discontinuation
  • 2000-01-27 EA EA200100808A patent/EA200100808A1/ru unknown
  • 2000-01-27 JP JP2000595968A patent/JP2002535383A/ja active Pending
  • 2000-01-27 BR BR0007754-2A patent/BR0007754A/pt not_active IP Right Cessation
  • 2000-01-27 NZ NZ512025A patent/NZ512025A/en unknown
  • 2000-01-27 WO PCT/US2000/001865 patent/WO2000044711A1/en not_active Application Discontinuation
  • 2000-01-27 IL IL14432100A patent/IL144321A0/xx unknown
  • 2000-01-27 AU AU26306/00A patent/AU769410B2/en not_active Ceased
  • 2000-01-27 CA CA002356345A patent/CA2356345A1/en not_active Abandoned
  • 2000-01-27 CZ CZ20012709A patent/CZ20012709A3/cs unknown
  • 2000-01-27 CN CN00803031A patent/CN1337944A/zh active Pending
  • 2000-01-27 EP EP00904570A patent/EP1147078A1/en not_active Withdrawn
  • 2000-01-27 HU HU0200605A patent/HUP0200605A3/hu unknown
• 2001
  • 2001-05-31 ZA ZA200104508A patent/ZA200104508B/xx unknown
  • 2001-07-24 NO NO20013639A patent/NO20013639D0/no not_active Application Discontinuation

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