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    • 专利摘要:
    An effective amount of a compound of formula (1) is administered to a patient to provide a useful method for inhibiting VCAM-1 in a patient and for treating chronic inflammatory conditions. <Formula 1> Where R 1 , R 2 , R 3 and R 4 are each independently a C 1 -C 6 alkyl group, Z is a thio, oxy or methylene group, A is a C 1 -C 4 alkylene group, R 5 is a C 1 -C 6 alkyl group or — (CH 2 ) n- (Ar), where n is an integer 0, 1, 2 or 3, and Ar is phenyl or hydroxy, methoxy, ethoxy, chloro, Naphthyl unsubstituted or substituted with one to three substituents selected from the group consisting of fluoro or C 1 -C 6 alkyl.
    公开号:KR20000065102A
    申请号:KR1019980708693
    申请日:1997-03-12
    公开日:2000-11-06
    发明作者:폴 에스. 라이트；스티븐 제이. 부쉬
    申请人:게리 디. 스트리트, 스티븐 엘. 네스비트；훽스트 마리온 로우셀, 인크.；
    IPC主号:
    专利说明:

    Inhibition of vascular cell adsorption molecule-1 with 2,6-di-alkyl-4-silyl-phenol and treatment of chronic inflammatory diseases
    Vascular cell adsorption molecule-1 (VCAM-1) and intercellular adsorption molecule-1 (ICAM-1) are cytokines such as interleukin-1 (IL-1), interleukin-4 (IL-4) and tumors Necrosis factor-α (TNF-α) is an adsorbent molecule in the class of immunoglobulins that is increased in vascular endothelial cells and smooth muscle cells. Through interaction with the appropriate integrin counter receptor, VCAM-1 and ICAM-1 mediate the adsorption and endothelial migration of leukocytes during the inflammatory response. Inhibitors of VCAM-1 and / or ICAM-1 have therapeutic uses for various forms of chronic inflammatory diseases, including asthma, rheumatoid arthritis, and autoimmune diabetes. For example, it is known that the expression of VCAM-1 and ICAM-1 increases in asthmatic patients [Pilewski, J. M. et al. Am. J. Pespir. Cell Mol. Biol. 12, 1-3 (1995); Ohkawara, Y. et al. Am. J. Pespir. Cell Mol. Biol. 12, 4-12 (1995) In addition, blocking the integrin receptors for VCAM-1 and ICAM-1 (VLA-1 and LFA-1, respectively) in the ovalbumin-sensitized rat model of allergic airway response Inhibits early and late phase reactions (Rabb, HA et al. Am. J. Respir.Care Med, 149, 1186-1191 (1994)).
    Moreover, VCAM-1 is included as a vehicle among other chronic inflammatory disorders such as rheumatoid arthritis and autoimmune diabetes. For example, in the vascular system of rheumatoid synovial membranes, expression of endothelial adsorption molecules including VCAM-1 is enhanced [Koch, A. E. et al., Lab. Invest. 64, 313-322 (1991); Morales-Ducret, J. et al., Immunol, 149, 1421-1431 (1996)]. Neutralizing antibodies directed against VCAM-1 and its counter receptors, VLA-4, can delay the onset of diabetes in mouse models in which the disease spontaneously develops (NOD mice) [Yang, X. D. et al. Proc. Natl. Acad. Sci. USA 90, 10494-10498 (1993). In addition, monoclonal antibodies against VCAM-1 may have beneficial effects in animal models of allograft rejection, suggesting that inhibitors of VCAM-1 expression may have utility in preventing transplant rejection [Orocz, Immunol from CG et al. Lett. 32, 7-12 (1992).
    VCAM-1 is expressed by cells as membrane bound and soluble forms, respectively. Soluble forms of VCAM-1 have been shown to induce chemotaxis of vascular endothelial cells in vitro and to promote angiogenic responses in rat corneas (Koch, A. E. et al. Nature 376, 517-519 (1995)). Inhibitors of soluble VCAM-1 expression are potent angiogenic components and have potential therapeutic utility in the treatment of diseases including tumor growth and metastasis [Folkman, J. and Shing, Y. J. Biol. Chem. 10931-10934 (1992).
    Accelerators for VCAM-1 and ICAM-1 create clones and are characterized by them. For example, each promoter contains a number of DNA sequence elements capable of binding the transcription factor, NF-kB [Iademarco, M. F. et al. J. Biol. Chem. 267, 16323-16329 (1992); Voraverger, G. et al. J. Immunol. 147, 2777-2786 (1991). The NF-kB group of transcription factors is important for the regulation of several genes that are extended within the site of inflammation. The activity of NF-kB as a transcription factor includes cleavage from the inhibitory subunit, IkB, in the cytoplasm. NF-kB subunits are translocated to the nucleus, bound to specific DNA sequence elements, and activate transcription of several genes, including VCAM-1 and ICAM-1 [Collins, T. Dung, Lab. Invest. 68, 499-508 (1993).
    It has been hypothesized that regulation of VCAM-1 gene expression can be paired with oxidative power through specific reduction-oxidation (redox) sensitive transcription or post-transcriptional regulatory factors. The antioxidants pyrrolidine dithiorocarbamate and N-acetylcysteine inhibit cytokine-induced expression of VCAM-1 in vascular endothelial cells, but not in ICAM-1 [Mauri, N. et al. J. Clin. Invest. 92. 1866-1874 (1993). This may indicate that inhibition of VCAM-1 expression by antioxidants includes some additional factors, but not the regulation of ICAM-1 expression.
    US Pat. No. 5,155,250, issued to Parker et al. On October 13, 1992, discloses 2,6-di-alkyl-4-silyl-phenols as anti-atherosclerosis. Furthermore, WO 95/15760, published June 15, 1995, discloses 2,6-di-alkyl-4-silyl-phenols as cerium cholesterol lowering agents.
    It may be beneficial to control the release of VCAM-1 and to treat VCAM-1 mediated efficacy. In addition, it may be beneficial to control or treat VCAM-1 inflammation without the associated side effects known to accompany the use of anti-inflammatory steroids and non-steroidal anti-inflammatory agents.
    Summary of the Invention
    It has been found that compounds corresponding to formula (1) can be used to inhibit cytokine-induced expression of VCAM-1.
    Where
    R 1 , R 2 , R 3 and R 4 are each independently a C 1 -C 6 alkyl group,
    Z is a thio, oxy or methylene group,
    A is a C 1 -C 4 alkylene group,
    R 5 is a C 1 -C 6 alkyl group or-(CH 2 ) n- (Ar), where n is an integer 0, 1, 2 or 3, and Ar is hydroxy, methoxy, ethoxy, chloro, fluoro Or phenyl or naphthyl unsubstituted or substituted with one to three substituents selected from the group consisting of C 1 -C 6 alkyl.
    Administering such compounds to a patient to inhibit VCAM-1; Inhibit or treat VCAM-1 mediated effects; VCAM-1 mediated inflammation can be inhibited or treated. This compound can be administered to inhibit or treat VCAM-1 mediated effects of diseases such as chronic inflammation, asthma, rheumatoid arthritis and autoimmune diabetes.
    <Brief Description of Drawings>
    FIG. 1 illustrates the effect of 2,6-di-t-butyl-4-[(dimethylphenylsilyl) methylthio] phenol (MDL 29353) on LPS induced VCAM-1 expression in rabbit aorta in vivo. Data are expressed as% aortic surface endothelial expressing VCAM-1 as measured by immunostaining with anti-rabbit VCAM-1 antibodies.
    As used herein, the term “C 1 -C 6 alkyl” means a saturated hydrocarbyl radical in the form of a straight, branched or cyclic ring of 1 to 6 carbon atoms. Included in this term are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, n-hexyl, cyclohexyl and the like.
    Likewise, the term “C 1 -C 4 alkylene” means a saturated hydrocarbyldiyl radical in straight or branched chain form of 1 to 4 carbon atoms. Included within the scope of this term are methylene, 1,2-ethane-diyl, 1,1-ethane-diyl, 1,3-propane-diyl, 1,2-propane-diyl, 1,3-butane-diyl, 1,4-butane-diyl and the like.
    When R 5 is a-(CH 2 ) n- (Ar) radical, the "-(CH 2 ) n- " residue represents a straight chain saturated hydrocarbyldiyl radical. "n" is defined as an integer 0, 1, 2 or 3. Thus, the residue "-(CH 2 ) n- " denotes a bond, methylene, 1,2-ethanediyl or 1,3-propanediyl. "-(Ar)-" residue represents an aryl radical defined as a substituted or unsubstituted phenyl or naphthal group. In these examples where the "-(Ar)-" residue is substituted aryl, phenyl or naphthyl may have 1 to 3 substituents at any position, or may be occupied by a hydrogen atom. Substituents are selected from the group consisting of hydroxy, methoxy, ethoxy, chloro, fluoro and C 1 -C 6 alkyl groups. Specifically included within the scope of the term "-(CH 2- (Ar)" are phenyl; naphthyl; phenylmethyl; phenylethyl; 3,4,5-trihydroxyphenyl; 3,4,5-trimeth Oxyphenyl; 3,4,5-triethoxyphenyl; 4-chlorophenyl; 4-methylphenyl; 3,5-di-t-butyl-4-hydroxyphenyl; 4-fluorophenyl; 4-chloro-1 -Naphthyl; 2-methyl-1-naphthylmethyl; 2-naphthylmethyl; 4-chlorophenylmethyl; 4-t-butylphenyl; 4-t-butylphenylmethyl and the like.
    Compounds of formula (1) may be prepared using processes and techniques known and understood by those of ordinary skill in the art. General synthetic methods for the preparation of compounds of formula (1) wherein Z is sulfur or oxygen are shown in Scheme (1), where all substituents are the same as described above unless stated otherwise.
    Generally, the phenols of formula (1a) are suitable 2,6-dialkyl-4-mers of formula (2) in suitable aprotic solvents such as dimethylformamide or dimethylacetamide or in aqueous solvents such as water / 2-butanone The captophenol or 2,6-dialkylhydroquinone (or appropriately protected derivative) is converted to a non-nucleophilic base such as sodium hydride, potassium carbonate or cesium carbonate, and a haloalkylene of appropriate formula (3) such as chloroalkylenesilane It can be prepared by reacting with silane.
    Starting materials for use in the general synthetic processes outlined in Scheme A are readily available to those skilled in the art. For example, certain phenolic starting materials for various compounds of formula (1) wherein Z is sulfur, such as 2,6-di-t-butyl-4-mercaptophenol, are described in US Pat. No. 3,576,883, US Pat. No. 3,952,064. No. 3,479,407 and Japanese Patent Application No. 73-28425. In addition, silyl starting materials for various compounds of formula (1) such as (trimethylsilyl) -methyl iodide, (trimethylsilyl) methyl bromide, (trimethylsilyl) methyl chloride, (1-chloropropyl) trimethylsilane are described in the literature. Synthesis 4, 318-9 (1988) and J. Am. Chem. Soc. 105, 5665-75 (1983). Further methods of preparing the appropriate silanes include the Grignard reaction, eg, 4-bromoanisole, with magnesium metal to prepare Grignard reagent, which is reacted with chlorodimethyl chloromethyl silane to give chloromethyldimethyl- Reactions such as making 4-methoxy phenyl silane.

    Alternatively, anisole may be reacted with Δ-butyllithium to be lithiated, and the prepared lithium compound is reacted with chlorodimethyl chloromethyl silane to produce chloromethyl dimethyl-2-methoxyphenyl silane.

    Where the 1-phenolic functional group of the compound of formula (2) can react with the compound of formula (3) under the reaction conditions, the 1-phenolic group of the compound of formula (2) is a standard phenol known and recognized in the art. Can be blocked with a blocking agent. The selection and use of specific circuit breakers are known to those of ordinary skill in the art. In general, the blocker should be selected for proper protection of the phenol in question during the subsequent synthesis step and should be readily removable under conditions that will not cause degradation of the desired product.
    Examples of suitable phenol protecting groups include ethers such as methoxymethyl, 2-methoxyethoxymethyl, tetrahydro-pyranyl, t-butyl and benzyl; Silyl ethers such as trimethylsilyl and t-butyldimethylsilyl; Esters such as acetates and benzoates; Carbonates such as methyl carbonate and benzyl carbonate; As well as sulfonates such as methanesulfonate and toluenesulfonate.
    In these examples where R 1 and R 2 are each t-butyl, the reaction of Scheme A may conveniently be carried out without blocking of the 1-phenol functional group.
    The following examples provide typical synthetic methods as described in Scheme A. These examples are to be understood as illustrative only and are not intended to limit the scope of the methods of the invention in any way. As used herein, designated terms have the following meanings; "g" represents grams, "bp" represents boiling points, "° C" represents degrees Celsius, "mm Hg" represents millimeters of mercury, "mp" represents melting points, and "mg" represents milligrams. "ΜM" represents micromolar and "μg" represents microgram.
    <Example 1>
    2,6-di-t-butyl-4-[(dimethylphenylsilyl) methylthio] phenol (MDL 29353)
    2,6-di-t-butyl-4-mercaptophenol (2.4 g, 10 mmol), potassium carbonate (1.4 g, 10 mmol), chloromethyldimethylphenylsilane (1.9 g, 10 mmol) and dimethylformamide ( 50 mL) were mixed and stirred overnight at room temperature under argon environment. The mixture was diluted with ice water and extracted with ethyl ether. The ether layer was washed with water, then brine, filtered through fluorosil-Na 2 SO 4 , and evaporated to yield orange oil (3.5 g). After the first distillation of the product (bp 160-170 ℃, 0.1 mm Hg ), was purified by silica gel chromatography (CCl 4: CHCl 3/1 : 1) to give were obtained the title compound as light yellow oil, which is to be solid white wax (2.3 g, 59%) was determined slowly.
    analysis
    Calcd for C 23 H 34 OSSi: C, 71.44; H, 8.86; S, 8.29;
    Found: C, 71.14; H, 8.86; S, 7.98.
    <Example 2>
    2,6-di-t-butyl-4-[(trimethylsilyl) methylthio] phenol (MDL 28,235)
    2,6-di-t-butyl-4-mercaptophenol (2.4 g, 10 mmol), potassium carbonate (1.4 g, 10 mmol), and dimethylacetamide (50 mL) were mixed and stirred at ambient temperature under argon environment It was. Chloromethyltrimethylsilane (1.3 g, 10 mmol) was added and stirred overnight. Warm up in the steam bath for 2 hours, cool and dilute with water. Extracted with ethyl ether, dried and evaporated until a light yellow solid (2.8 g) was obtained and recrystallized (CH 3 CN) to give 1.1 g (34%) of the title compound; Melting point 100-101 ° C.
    analysis
    Calcd for C 18 H 32 OSSi: C, 66.60; H, 9.88; S, 9.88;
    Found: C, 66.83; H, 10.05; S, 9.91.
    <Example 3>
    2,6-dimethyl-4-[(trimethylsilyl) methyloxy] phenol
    2,6-dimethylhydroquinone (1.4 g, 10 mmol), potassium carbonate (1.4 g, 10 mmol), chloromethyltrimethylsilane (1.9 g, 10 mmol) and dimethylformamide (50 mL) were mixed. Stir at room temperature under inert environment until reaction is complete. The mixture was diluted with ice water and extracted with ethyl ether. The ether layer was washed with water, then brine and filtered through fluorosil-Na 2 SO 4 . Evaporation gave the title compound which was purified by silica gel chromatography.
    <Example 4>
    2,6-di-t-butyl-4-[(4-chlorophenyldimethylsilyl) methyloxy] phenol (MDL 104,280)
    2,6-di-t-butylbenzhydroquinone (13.7 g, 61.6 mM), potassium carbonate (9.4 g, 68 mM), chloromethyl (4-chlorophenyl) dimethyl silane (14.9 g, 68 mM) and catalytic amount of iodide Potassium was refluxed for 3 days under N 2 in acetonitrile (200 ml). The solid was filtered off and evaporated. Redissolved in ethyl acetate and washed with water, then brine, dried over anhydrous magnesium sulfate, filtered and evaporated. The resulting orange oil was distilled to 135 ° C. @ 0.1 mm Hg to remove impurities at lower boiling points, and then the product could be purified by distillation (bp ° C. @ 0.1 mm Hg). On standing, the crystallized product was recrystallized from hexane to give fine white acicular crystals (7.4 g, 27% yield); mp 102-105 ° C.
    analysis
    Calcd for C 23 H 33 ClO 2 Si: C, 68.20; H, 8.21
    Found: C, 68.39; H, 8.13
    NMR (CDCl 3 ): 7.53 (d, 2H, J 8.3), 7.34 (d, 2H, J 8.3), 6.79 (s, 2H), 4.73 (s, 1H), 3.71 (s, 2H), 1.42 (s , 18H), 0.41 (s, 6H).
    Example 5
    2,6-di-t-butyl-4-[(dimethyl-4-fluorophenylsilyl) methyloxy] phenol (MDL 104,389)
    2,6-di-t-butylbenzhydroquinone (10.0 g, 45 mM), potassium carbonate (6.2 g, 45 mM) and dimethyl (4-fluorophenyl) iodomethylsilane (13.2 g, 45 mM) It was refluxed for 3 days under N 2 in nitrile (150 ml). The solid was filtered off and evaporated. It was redissolved in ethyl acetate and washed with water, then with brine, dried over anhydrous magnesium sulfate, filtered and evaporated until left to crystallize very pale yellow oil. This material was recrystallized from methanol to give a white crystalline solid (5.9 g, 34% yield). mp 90-93 ° C.
    analysis
    Calcd for C 23 H 33 FO 2 Si: C, 71.09; H, 8.86
    Found: C, 70.96; H, 8.58
    NMR (CDCl 3 ): 7.58 (dd, 2H, J 8.5, 6.2), 7.10-7.04 (m, 2H), 6.80 (s, 2H), 4.73 (s, 1H), 3.71 (si, 2H), 1.43 ( s, 18 H), 0.41 (s, 6 H).
    <Example 6>
    2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol (MDL 103,902)
    2,6-di-t-butylbenzhydroquinone (5.43 g, 24.4 mM), potassium carbonate (3.7 g, 26.8 mM), and dimethyl (iodomethyl) phenylsilane (7.4 g, 26.8 mM) were acetonitrile (125 in reflux overnight under N 2 . The solid was filtered off and evaporated. Redissolved in ethyl acetate and washed with water, then brine, dried over anhydrous magnesium sulfate, filtered and evaporated. The resulting orange oil was distilled to 135 ° C @ 0.1 mm Hg to remove impurities of lower boiling point, and then the product (bp 155-165 ° C @ 0.1 mm Hg) could be purified by distillation. On standing, the crystallized product was recrystallized from methanol to give a white solid (5.8 g, 64% yield); mp 82-83 ° C.
    analysis
    Calcd for C 23 H 34 0 2 Si: C, 74.54; H, 9.25
    Found: C, 74.51; H, 9.28
    NMR (CDCl 3 ): 7.64-7.58 (m, 2H), 7.42-7.32 (m, 2H), 6.80 (s, 2H), 4.72 (s, 1H), 3.73 (s, 2H), 1.42 (s, 18H ), 0.42 (s, 6H).
    <Example 7>
    2,6-di-t-butyl-4-[(dimethyl-4-methoxyphenylsilyl) methyloxy] phenol (MDL 105,074)
    2,6-di-t-butylbenzhydroquinone (13.7 g, 61.6 mmol), potassium carbonate (9.4 g, 68 mmol), chloromethyl (dimethyl) -4-methoxyphenylsilane (14.6 g, 68 mmol) and catalytic amount Potassium iodide was refluxed in acetonitrile (200 ml) under N 2 for 3 days. The solid was filtered off and evaporated. Redissolved in ethyl acetate and washed with water, then brine, dried over anhydrous magnesium sulfate, filtered and evaporated. The resulting orange oil was distilled to 135 ° C @ 0.1 mm Hg to remove impurities of lower boiling point, and then the product (bp 155-165 ° C @ 0.1 mm Hg) could be purified by distillation. On standing the crystallized product was recrystallized from hexane to give a white solid (4.9 g, 19% yield); mp 122-123 ° C.
    analysis
    Calcd for C 24 H 36 0 3 Si: C, 71.95; H, 9.06
    Found: C, 71.80; H, 9.00
    NMR (CDCl 3 ): 7.53 (d, 2H, J 8.6), 6.93 (d, 2H, J 8.6), 6.80 (s, 2H), 4.71 (s, 1H), 3.81 (s, 3H), 3.70 (s , 2H), 1.42 (s, 18H), 0.39 (s, 6H).
    <Example 8>
    2,6-dimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol (MDL 103,719)
    2,6-dimethylhydroquinone (10.0 g, 72.4 mmol M), potassium carbonate (10.0 g, 72.4 mmol), dimethyl (chloromethyl) phenylsilane (13.4 g, 72.4 mmol) in acetonitrile (150 ml) under argon for 72 hours Reflux for a while. The mixture was cooled, diluted with water and extracted with ether. The oil was distilled at 145 to 160 ° C. @ 0.1 mm Hg to yield 4.9 g of a light yellow oil.
    analysis
    Calcd for C 16 H 22 0 2 Si: C, 71.28; H, 7.74
    Found: C, 71.27; H, 7.74
    Example 9
    2-t-butyl-6-methyl-4-[(dimethylphenylsilyl) methylthio] phenol (MDL 104,518)
    2-t-butyl-6-methyl-4-mercaptophenol (11.8 g, 60.1 mmol), potassium carbonate (6.0 g, 11.8 mmol), dimethyl (chloromethyl) phenylsilane (11.1 g, 60.1 mmol) under argon Reflux in acetonitrile (150 ml) for 24 hours. The mixture was cooled, diluted with water and extracted with ether. The ether layer was evaporated to dryness to yield 21.9 g of oil. The oil was distilled at 145 to 160 ° C. @ 0.1 mm Hg to yield 4.9 g of a light yellow oil.
    analysis
    Calcd for C 20 H 28 OSSi: C, 69.71; H, 8.19
    Found: C, 69.76; H, 8.20
    <Example 10>
    2,6-di-t-butyl-4-[(dimethyl-2-methoxyphenylsilyl) methyloxy] phenol (MDL 104,036)
    A mixture of chloromethyldimethyl (2-methoxy) phenylsilane (27.2 g, 0.127 mol), sodium iodide (19 g, 0.127 mol) and acetonitrile (350 ml) was heated at reflux for 28 hours. The mixture was cooled to ambient temperature and 2,6-di-t-butyl-1,4-hydroquinone (31.5 g, 0.14 mol) and potassium carbonate (20.8 g, 0.15 mol) were added. The mixture was refluxed for 7 days under nitrogen environment. The mixture was cooled, poured into water (400 ml) and ethyl acetate (400 ml) and the organic layer was separated. The organic layer was evaporated and the residue was chromatographed on silica gel (hexane / ethyl acetate 9/1). The chromatographed product was recrystallized (methanol) to give the product (15.6 g 31%) as a white solid; mp 89-90 ℃
    analysis
    Calcd for C 24 H 36 0 3 Si: C, 71.95; H, 9.06
    Found: C, 71.84; H, 9.05
    <Example 11>
    2,6-di-t-butyl-4-[(dimethyl-2,5-dimethoxyphenylsilyl) methyloxy] phenol (MDL 103,016)
    The product was obtained as a white solid using chloromethyl dimethyl-2,5-dimethoxy-phenylsilane (14 g, 57 mmol) as the silane as in the above process for preparing the compound; mp 103-104 ℃
    analysis
    Calcd for C 25 H 38 O 4 Si: C, 69.72; H, 8.89
    Found: C, 69.71; H, 8.72
    <Example 12>
    2,6-di-t-butyl-4-[(dimethyl-2,3-dimethoxyphenylsilyl) methyloxy] phenol (MDL 108,750)
    The product was obtained as a white solid using chloromethyl (dimethyl) -2,3-dimethoxy phenylsilane (11.3 g, 46 mmol) as the silane as in the above process for preparing the compound; mp 94.5-96 ℃
    analysis
    Calcd for C 25 H 38 O 4 Si: C, 69.72; H, 8.89
    Found: C, 69.84; H, 8.91
    Example 13
    2,6-di-t-butyl-4-[(dimethyl-4-t-butylphenylsilyl) methyloxy] phenol (MDL 106,630)
    The product was obtained as a white solid using 4-t-butylphenyl chloromethyl dimethylsilane (6.2 g, 25.7 mmol) as silane as in the above process for preparing the compound; mp 114-115 ℃
    analysis
    Calcd for C 27 H 42 0 2 Si: C, 76.00; H, 9.92
    Found: C, 75.94; H, 1013
    <Example 14>
    2,6-di-t-butyl-4-[(benzyldimethylsilyl) methyloxy] phenol (MDL 107,411)
    The product was obtained as a white solid using benzyl chloromethyl dimethyl silane (7.13 g, 35.9 mmol) as silane as in the method for preparing the above-mentioned compound; mp 76-77 ℃
    analysis
    Calcd for C 24 H 36 0 2 Si: C, 74.95; H, 9.43
    Found: C, 74.94; H, 9.36
    <Example 15>
    2,6-di-t-butyl-4-[(dimethyl-p-dimethoxybenzylsilyl) methyloxy] phenol (MDL 103,816)
    Step a; Preparation of Dimethyl-p-methoxybenzyl-chloromethylsilane: Magnesium turning (9.7 g, 0.4 g atoms) was stirred overnight under nitrogen using a Teflon® paddle. This "activated" magnesium was suspended in dry THF (100 mL) and iodine crystals were added. To this suspension was added a solution of p-methoxy-benzylbromide (80.8 g, 0.4 mol) in THF (400 mL) at a rate to maintain mild reflux. After the addition was complete, stirring was continued until almost all magnesium was consumed (˜2 hours). A solution of dimethylchloromethylchlorosilane (52.7 mL, 0.4 mol) in THF (200 mL) was added dropwise and the mixture was stirred at room temperature overnight. The reaction mixture was quenched with saturated aqueous ammonium chloride (500 mL) and stirred at room temperature (˜2 hours). The precipitated magnesium salt was filtered off and diluted with ether (300 mL). The organic phase was separated, washed with water (3 × 250 mL), saturated aqueous sodium chloride (3 × 250 mL), dried over anhydrous magnesium sulfate, filtered and evaporated. The resulting brown oil was distilled off and purified to afford the title compound. Yield 55% at 5 mm Hg, bp 110-115 ° C.
    analysis
    Calcd for C 10 H 15 ClOSi: C, 55.93; H, 7.15
    Found: C, 55.40; H, 7.15
    Step b; Preparation of 2,6-di-t-butyl-4-[(dimethyl-p-methoxybenzyl-silyl) methyloxy] phenol (MDL 108,816): Dimethyl-p-methoxybenzyl in acetonitrile (250 mL) A mixture of chloromethylsilane (28 g, 0.13 mmol), sodium iodide (0.5 g, cat), 2,6-di-t-butybenzhydroquinone (23 g, 0.1 mol) and cesium carbonate (32 g, 0.1 mol) Was heated at reflux for 6 days, cooled and poured into a mixture of water / ethyl acetate (400 mL each). The organic layer was separated, dried, evaporated and the residue was heated on a Kugelrohr apparatus for 3 hours at 110 ° C. and another 2 hours at 140-160 ° C. The residue was left to solidify to give the product as a wax solid (20.9 g, 39%); mp 58-60 ° C.
    analysis
    Calcd for C 25 H 38 O 3 Si: C, 72.41; H, 8.87
    Found: C, 70.29; H, 8.96
    The following compounds can be prepared in a similar manner as described in Examples 1-14 above.
    2,6-di-t-butyl-4-[(triethylsilyl) methylthio] phenol
    2,6-di-t-butyl-4-[(diethylphenylsilyl) methylthio] phenol
    2,6-di-t-butyl-4-[(tripropylsilyl) methylthio] phenol
    2,6-di-t-butyl-4-[(dipropylphenylsilyl) methylthio] phenol
    2,6-di-t-butyl-4-[(triisopropylsilyl) methylthio] phenol
    2,6-di-t-butyl-4-[(diisopropylphenylsilyl) methylthio] phenol
    2,6-di-t-butyl-4-[(tributylsilyl) methylthio] phenol
    2,6-di-t-butyl-4-[(dibutylphenylsilyl) methylthio] phenol
    2,6-di-t-butyl-4-[(triisobutylsilyl) methylthio] phenol
    2,6-di-t-butyl-4-[(diisobutylphenylsilyl) methylthio] phenol
    2,6-di-t-butyl-4-[(tri-t-butylsilyl) methylthio] phenol
    2,6-di-t-butyl-4-[(di-t-butylphenylsilyl) methylthio] phenol
    2,6-di-methyl-4-[(trimethylsilyl) methylthio] phenol
    2,6-di-methyl-4-[(dimethylphenylsilyl) methylthio] phenol
    2,6-di-methyl-4-[(dibutylphenylsilyl) methylthio] phenol
    2,6-di-methyl-4-[(tri-t-butylsilyl) methylthio] phenol
    2,6-di-methyl-4-[(di-t-butylphenylsilyl) methylthio] phenol
    2,6-di-ethyl-4-[(trimethylsilyl) methylthio] phenol
    2,6-di-ethyl-4-[(dimethylphenylsilyl) methylthio] phenol
    2,6-di-ethyl-4-[(tri-t-butylsilyl) methylthio] phenol
    2,6-di-ethyl-4-[(di-t-butylphenylsilyl) methylthio] phenol
    2,6-di-propyl-4-[(trimethylsilyl) methylthio] phenol
    2,6-di-propyl-4-[(dimethylphenylsilyl) methylthio] phenol
    2,6-di-isopropyl-4-[(trimethylsilyl) methylthio] phenol
    2,6-di-isopropyl-4-[(dimethylphenylsilyl) methylthio] phenol
    2,6-di-butyl-4-[(trimethylsilyl) methylthio] phenol
    2,6-di-butyl-4-[(dimethylphenylsilyl) methylthio] phenol
    2,6-dimethyl-4-[(trimethylsilyl) methyloxy] phenol
    2,6-dimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol
    2,6-dibutyl-4-[(triethylsilyl) methyloxy] phenol
    2,6-dibutyl-4-[(diethylphenylsilyl) methyloxy] phenol
    2,6-di-t-butyl-4-[(triethylsilyl) methyloxy] phenol
    2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol
    A general synthetic scheme for preparing compounds of formula (1) wherein Z is methylene is in Scheme B and all substituents are as described above unless otherwise indicated.
    In general, the phenols of formula (1b) can be prepared according to Scheme (B) in a two-step process. In step (a), a suitable haloalkylenesilane of formula (3) is reacted with magnesium metal in a suitable aprotic solvent such as ethyl ether to form a magnesium halide salt. The magnesium halide salt (Grignard reagent) was then reacted with the appropriate 3,5-dialkyl-4-hydroxybenzaldehyde (or suitably protected derivative) of formula (4) to prepare an alcohol of formula (5). In step (b), the alcohol of formula (5) can be reduced to the desired phenol of formula (1b) by various reduction techniques and processes as known and understood in the art. For example, the alcohol of formula (5) can be reduced by reacting it with sodium in liquid ammonia using Birch reduction.
    Starting materials for use in the general synthetic processes outlined in Scheme B are readily available or can be readily prepared according to standard techniques or processes. If it is necessary to prevent undesired side reactions, the 3,5-dialkyl-4-hydride of formula (4) in Scheme (B) prior to Grignard reaction with standard phenol blockers as described in Scheme A above It is possible to block the 1-phenol functionality of oxy-benzaldehyde.
    The following examples provide typical synthetic methods as described in Scheme B. It should be understood that this embodiment is exemplary only and is not intended to limit the scope of the method of the present invention in any case.
    <Example 16>
    2,6-dimethyl-4- [2- (trimethylsilyl) ethyl] phenol
    Step a: Magnesium turning (240 mg, 10 mmol) and anhydrous ethyl ether were mixed under inert environment. A solution of chloromethyltrimethylsilane (1.9 g, 10 mmol) in anhydrous ethyl ether was added. Stir until magnesium metal is dissolved. A solution of 3,5-dimethyl-4-hydroxybenzaldehyde (1.5 g, 10 mmol) in anhydrous ethyl ether was added. Stir until the reaction is complete. The reaction mixture was cooled to 0 ° C and saturated ammonium chloride solution was added. The ether layer was separated, washed with water and dried (MgSO 4 ). It was evaporated to prepare 4-hydroxy-3,5-dimethyl-α-[(trimethylsilyl) -methyl] benzenemethanol and purified by silica gel chromatography.
    Step b: Sodium metal (520 mg, 22.6 mmol) and liquid ammonia (13 mL) were mixed. To this solution was added dropwise a solution of 4-hydroxy-3,5-dimethyl-α-[(trimethylsilyl) -methyl] benzenemethanol (2.22 g, 10 mmol) in ethyl alcohol (0.5 g) and ethyl ether (5 mL). It was. After the blue disappeared, water (13 mL) was added carefully, extracted with ethyl ether, dried (MgSO 4 ) and the solvent evaporated. The residue was purified by silica gel chromatography to give the title compound.
    Alternatively, all substituents may be prepared for compounds of formula (1) in which Z is methylene according to the process described in Scheme C as described above unless otherwise indicated.
    In general, the phenol of formula (1b) may be prepared by first reacting a magnesium metal with a suitable haloalkylenesilane of formula (3) in an aprotic solvent such as a suitable ethyl ether to form a magnesium halide salt. The magnesium halide salt (Grignard reagent) can then be reacted with the appropriate 3,5-dialkyl-4-hydroxy-benzyl halide of formula (6) to produce the desired phenol of formula (1b).
    Starting materials for use in the general synthetic processes outlined in Scheme C are readily available or can be readily prepared according to standard techniques or processes. For example, the preparation of 3,5-dimethyl-4-acetoxy-benzylbromide is described in Tetrahedron 33, 3097-103 (1977). 3,5-dimethyl-4-acetoxy-benzylbromide can be converted to the corresponding phenol starting material by standard hydrolysis processes.
    If undesired side reactions are desired, 1 of 3,5-dialkyl-4-hydroxy benzyl halide of formula (6) in Scheme C prior to Grignard reaction with a standard phenol blocker as described in Scheme A above Can block phenolic functionalities;
    The following examples provide the typical synthetic methods described in Scheme C. It is to be understood that this embodiment is exemplary only, and is not intended to limit the scope of the method of the present invention in any case.
    <Example 17>
    2,6-dimethyl-4- [2- (trimethylsilyl) ethyl] phenol
    Magnesium turning (240 mg, 10 mmol) and anhydrous ethyl ether were mixed under inert environment. A solution of chloromethyltrimethylsilane (1.9 g, 10 mmol) in anhydrous ethyl ether was added. Stir until magnesium metal is dissolved. A solution of 4-bromomethyl-2,6-diethylphenol (2.43 g, 10 mmol) in anhydrous ethyl ether was added and the mixture was refluxed until the reaction was complete. This was poured onto a mixture of ice / hydrochloric acid and the layers separated. The ether layer was washed with water, dried (MgSO 4 ) and evaporated to afford the title compound which was purified by silica gel chromatography.
    The following compounds can be prepared using methods similar to those in Example 16 described above.
    2,6-dipropyl-4- [2- (trimethylsilyl) ethyl] -phenol
    2,6-dipropyl-4- [2- (dimethylphenylsilyl) ethyl] -phenol
    2,6-diisopropyl-4- [2- (trimethylsilyl) ethyl] -phenol
    2,6-diisopropyl-4- [2- (dimethylphenylsilyl) ethyl] -phenol
    2,6-diisobutyl-4- [2- (trimethylsilyl) ethyl] -phenol
    2,6-diisobutyl-4- [2- (dimethylphenylsilyl) ethyl] -phenol
    2,6-dibutyl-4- [2- (trimethylsilyl) ethyl] -phenol
    2,6-dibutyl-4- [2- (dimethylphenylsilyl) ethyl] -phenol
    2,6-di-t-butyl-4- [2- (trimethylsilyl) ethyl] -phenol
    2,6-di-t-butyl-4- [2- (dimethylphenylsilyl) ethyl] -phenol
    2,6-di-t-butyl-4- [2- (tri-t-butylsilyl) ethyl] -phenol
    2,6-di-t-butyl-4- [2- (di-t-butylphenylsilyl) ethyl] -phenol
    2,6-dimethyl-4- [2- (trimethylsilyl) ethyl] -phenol
    2,6-dimethyl-4- [2- (dimethylphenylsilyl) methyl] -phenol
    It is understood that the compounds of formula (1) exist in various stereoisomeric forms. All stereoisomeric forms corresponding to the above formulas are intended to be included within the scope of the present invention as described according to standard practice for expressing stereoisomeric structures.
    Compounds of formula (1), for example 2,6-di-alkyl-4-silyl-phenol, are known in the art. In particular, compounds of formula (1) are described in US Pat. No. 5,155,250. Preferred compounds of formula (1) are those in which R 1 and R 2 are C 4 alkyl groups, R 3 and R 4 are C 1 alkyl groups, A is a C 1 alkylene group, and R 5 is-(CH 2 ) n- (Ar Wherein n is 0 and Ar is phenyl unsubstituted or substituted with 1 to 3 substituents selected from the group consisting of hydroxy, methoxy, ethoxy, chloro, fluoro or C 1 -C 6 alkyl) It is the ones that are. More preferred compound is 2,6-di-t-butyl-4 [(dimethylphenylsilyl) methyl] -thio-phenol.
    As used herein, the term “patient” relates to a warm blood-animal or mammal infected with a particular VCAM-1 mediated inflammatory disease. Primates, including guinea pigs, dogs, cats, rats, mice, hamsters, rabbits and humans, are understood to be examples of patients within the meaning of this term.
    The term "chronic inflammatory disease" relates to a disease or condition characterized by inflammation that persists in the absence of identifiable stimulants or bacterial pathogens. Inflammatory diseases for which treatment with the compound of formula (1) would be particularly useful include asthma, chronic inflammation, rheumatoid arthritis, autoimmune diabetes, transplant rejection and tumor angiogenesis. Particularly preferred compounds of formula (1) for the treatment of inflammatory diseases in patients are
    2,6-di-t-butyl-4-[(dimethylphenylsilyl) methylthio] phenol
    2,6-di-t-butyl-4-[(trimethylsilyl) methylthio] phenol
    2,6-di-t-butyl-4-[(4-chlorophenyldimethylsilyl) methyloxy] phenol
    2,6-di-t-butyl-4-[(dimethyl-4-fluorophenylsilyl) methyloxy] phenol
    2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol
    2,6-di-t-butyl-4-[(dimethyl-4-methoxyphenylsilyl) methyloxy] phenol
    2,6-dimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol
    2-t-butyl-6-methyl-4-[(dimethylphenylsilyl) methylthio] phenol
    2,6-di-t-butyl-4-[(dimethyl-2-methoxyphenylsilyl) methyloxy] phenol
    2,6-di-t-butyl-4-[(dimethyl-2,5-dimethoxyphenylsilyl) methyloxy] phenol
    2,6-di-t-butyl-4-[(dimethyl-2,3-dimethoxyphenylsilyl) methyloxy] phenol
    2,6-di-t-butyl-4-[(dimethyl-4-t-butylphenylsilyl) methyloxy] phenol
    2,6-di-t-butyl-4-[(benzyldimethylsilyl) methyloxy] phenol
    It includes.
    A “therapeutically effective amount” of a compound of formula (1) is an amount effective to alleviate the signs associated with an inflammatory disease upon single or multiple dose administration to a patient. An “vascular cell adsorption molecule-1 inhibitory effective amount” of a compound of formula (1) is an amount effective to alleviate the signs associated with vascular cell adsorption molecule-1 mediated status upon single or multiple dose administration to a patient. As used herein, "relieving symptoms" of an inflammatory disease or vascular cell adsorption molecule-1 mediated condition means reducing the anticipated exacerbation if not treated and does not necessarily indicate complete elimination or treatment of the disease. Do not. In addition, alleviation of symptoms includes prophylaxis.
    In determining the therapeutically effective amount or dosage, a number of factors are contemplated by the diagnosis of the attending physician, including species of mammals; Its size, age, and general state of health; Certain diseases; The degree or relevance of the disease; Individual patient response; The specific compound to be administered; Mode of administration; Bioavailability of the administered agent; Selected method of administration; The use of accompanying drugs; And other related environments.
    The therapeutically effective amount of the compound of formula (1) will generally vary from about 1 mg / kg to about 5 g / kg per day. A daily dosage of about 1 mg / kg to about 500 mg / kg is preferred. Likewise, the vascular cell adsorption molecule-1 inhibitory effective amount of the compound of formula (1) will generally vary from about 1 mg / kg to about 5 g / kg per day. A daily dosage of about 1 mg / kg to about 500 mg / kg is preferred.
    Compounds of the invention are inhibitors of VCAM-1 expression. The compounds of the present invention are believed to exert their inhibitory effects through inhibition of VCAM-1 increase by cytokines, thereby preventing or alleviating the signs of inflammatory diseases including asthma, chronic inflammation, rheumatoid arthritis, autoimmune diabetes, and the like. . However, it is understood that the present invention is not limited by any particular theory and suggests a mechanism for explaining its efficiency in end use.
    In effective treatment of a patient, the compound of formula (1) may be administered in any form or manner, including oral and parenteral routes, which make it effective bioavailable. For example, the compound can be administered orally, subcutaneously, intramuscularly, intravenously, transdermally, intranasally, rectally and the like. Oral administration is generally preferred. Those skilled in the pharmaceutical industry can easily select the appropriate form and mode of administration depending on the disease state to be treated, the stage of the disease, and other relevant circumstances. Remington's Pharamaceutical Sciences, 18th Edition, Mack Publishing Co. (1980)].
    The compound of formula (1) may be administered in the form of a pharmaceutical composition or medicament, which is prepared by combining the compound of formula (1) with a pharmaceutically acceptable carrier or excipient, the proportions and characteristics of which are selected And standard pharmaceutical experiments.
    Pharmaceutical compositions or medicaments have been prepared in a manner known in the pharmaceutical art. The carrier or excipient can be a solid, semi-solid, or liquid substance, which can act as a carrier or medium of the active ingredient. Suitable carriers or excipients are known in the art. The pharmaceutical compositions can be used orally or parenterally and can be administered to the patient in the form of tablets, capsules, suppositories, solutions, suspensions and the like.
    Pharmaceutical compositions can be administered orally, for example, with an inert diluent or an edible carrier. These may be packaged in gelatin capsules or included into tablets. For therapeutic oral administration, the compound of formula (1) can be mixed with excipients and used in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, chewing gums and the like. These preparations should contain at least 4% by weight of the compound of formula (1), the active ingredient, but can be modified according to the particular form, advantageously from 4 to about 70% by weight of the unit. A unit dosage form suitable for administration in the amount of active ingredient present in the composition will be obtained.
    In addition, tablets, pills, capsules, troches, and the like may include one or more of the following adjuvants: binders such as microcrystalline cellulose, gum tragacanth or gelatin; Excipients such as starch or lactose; Disintegrants (eg, alginic acid, primogel, corn starch, etc.); Lubricants such as magnesium stearate or sterotex; Glidants such as colloidal silicon dioxide; And sweetening agents such as sucrose or saccharin, or flavoring agents such as peppermint, methyl silicate or orange flavoring and the like. When the dosage unit form is a capsule, it may contain a liquid carrier, such as polyethylene glycol or fatty oil, in addition to the material of that form. Other dosage unit forms may include various other materials that modify the physical form of the dosage unit, such as, for example, a coating. Thus, tablets or pills may be coated with sugar, shellac, or other enteric coatings. Syrups, in addition to the active ingredient, may contain sucrose and certain preservatives, dyes and colorants and flavors as sweeteners. The materials used to prepare these various compositions must be pharmaceutically pure and nontoxic in the amounts used.
    For parenteral administration, the compounds of formula (1) may be mixed in solution or suspension. These formulations should comprise at least 0.1% of a compound of the present invention, but may vary from 0.1 to about 50% by weight. Appropriate dosages will be obtained with the amounts of active ingredient present in such compositions.
    In addition, solutions or suspensions may contain one or more of the following auxiliaries, depending on the solubility and other properties of the compound of formula (1): sterile diluents, for example water for injection, physiological saline, fixed oils, polyethylene glycols, glycerin, Propylene glycol or other synthetic solvents; Antibacterial agents such as benzyl alcohol or methyl parabens; Antioxidants such as ascorbic acid or sodium bisulfite; Chelating agents such as ethylene diaminetetraacetic acid; Buffers such as acetate, citrate or phosphate; And toxic titrants such as sodium chloride or dextrose. Parenteral preparations can be placed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.
    Example 18
    Cell Surface ELISA for VCAM-1 / ICAM-1
    96-well plates in 100 μL medium per well of proliferating human umbilical vein endothelial cells (HUVEC) or human aortic smooth muscle cells (HASMC) from Clonetics (San Diego, CA) at 20,000 cells per cm 2 Cultured on Cultures were maintained for 2 days in growth medium (EGM or SMGM2, Clonetics, San Diego, Calif.) Before adding cytokines or drugs. Cytokine ± compounds were added for 20 to 24 hours before analyzing the adsorption molecule levels. Tumor necrosis factor (Genzyme, Cambridge, Mass.) Was added to the culture at 500-1000 units / mL. Interleukin-4 (GIBCO-BRL, Gaithersbug, MD) was added to the culture at 100-200 pg / mL (100 μL of serially diluted Cytokine + Compound diluted in individual 96-well plates was transferred to the plate containing cells. Culture medium was not exchanged before adding effector material). The culture medium was removed and the monolayers were washed twice with Hanks buffered saline (HBSS) at room temperature. Primary antibody (anti-human VCAM-1 from Upstate Biotechnology, Inc., Lake Placid, NY, or anti-human ICAM-1 from Immunotech, Inc., Westbrook, ME) was added to each well (1 in HBSS) μg + 5% neonatal calf serum, GIBCO-BRL, Gaithersburg, MD) was incubated at 37 ° C. for 1 hour. The wells were washed twice with HBSS, and then each well was diluted with a 1/1000 dilution of goat anti-mouse IgG bound to horse radish peroxidase (BioRad, Hercules, CA) in HBSS + 5% newborn calf serum. 100 μL was added and incubated at 37 ° C. for 1 hour. After washing the wells three times with HBSS, 100 μL of TMB substrate (BioRad, Hercules, CA) was added to each well. After the blue color appeared, 50 μL of 1 NH 2 SO 4 was added to stop the reaction. Absorbance was measured using a plate reader at 450 nm. IC 50 values were determined from curves of absorbance values obtained from dilutions of continuous compounds (dissolved in dimethyl sulfoxide).
    IC 50 values are defined as drug concentrations that inhibit the expression of cytokine-induced adsorption molecules of 50%. The induction level was determined by subtracting the maximum value of adsorption molecule expression in the cytokine-induced culture from the baseline level of adsorption molecule expression in the culture (-cytokine). VCAM-1 is typically induced 5-7 fold. ICAM-1 is typically induced 5-10 fold. Each drug concentration was tested four times in the wells. The single point test at 50 μM was evaluated as described for the IC 50 measurement, except for the fact that the data indicate the level of inhibition without correction for basal expression (base adsorption molecule expression was determined by 10-20%).
    Table 1 summarizes the ability of various compounds of the invention to inhibit VCAM-1 using human aortic smooth muscle cells (HASMC). In this experiment, cells were co-cultured with interleukin-4 and the listed compounds for about 20 hours before assessing cell surface VCAM-1 levels. Each column represents an individual experiment.
    TABLE 1
    Compound Number (MDL Number)HSMC-1 (% inhibition, 50 μM)HSMC-2 (% inhibition, 50 μM)HSMC-3 (% inhibition, 50 μM)VCAM-1 (average) 28,2357.818.042.022.6 29,35358.060.046.054.7 103,719 49.042.045.5 103,90249.060.043.050.7 104,28054.763.044.053.9 104,5184.447.026.025.8 105,07428.452.047.042.4
    Table 2 summarizes the ability of various compounds of the invention to selectively inhibit VCAM-1 or to inhibit VCAM-1 and ICAM-1, respectively, using proliferated human venous endothelial cells (HUVEC). In this experiment, cells were co-cultured with tumor necrosis factor-α with designated compounds for about 20 to 24 hours before assessing cell surface adsorption molecule expression.
    TABLE 2
    Compound Number (MDL Number)VCAM-1 (% inhibition, 50 μM) * ICAM-1 (% inhibition, 50 μM) @28,2354.3(3.0) 29,3538.76.0 103,71917.077.0 103,90225.339.0 104,28027.322.0 104,518(1.0)78.0 105,07420.0(8.0)* 3 averages @ 2 averages
    Table 3 illustrates the activity of selected compounds when eight series of drug dilutions in vascular endothelial and smooth muscle cells were tested for cytokine induction of VCAM-1 expression. In addition, subtraction of basal VCAM-1 expression was used for the calculation of IC 50 values. In addition, each compound was tested in a similar manner as in ICAM-1 inhibition. No inhibition of ICAM-1 was detected in vascular smooth muscle cells (up to 100 μM). In vascular endothelial cells, only MDL 103, 902 at 50 and 100 μM showed significant inhibition of ICAM-1 expression, which could be calculated by the loss of cell adsorption on the tissue culture surface as observed microscopically. .
    TABLE 3
    Compound Number (MDL Number)VCAM-1, HUVEC (IC 50 , μm)VCAM-1, HSMC (IC 50 , μm) 29,3531910 103,902115 105,0741240
    Example 19
    In Vivo Inhibition of VCAM-1 Increase by MDL 29353 in Rabbit Aorta
    New Zealand white rabbits were fed ± 0.4% MDL 29353 diet for 3 weeks before lipopolysaccharide (LPS, 40 μg / animal, intravenous ear vein). LPS was injected and after 4 hours the aorta was removed from each animal and washed briefly in phosphate buffered saline. Tissues were incubated overnight with RB1 / 9 antibody (mouse anti-rabbit VCAM-1) at 4 ° C. to perform immunohistochemistry of VCAM-1. Bound RB1 / 9 was detected using biotinylated goat anti-mouse secondary antibody (60 minutes, RT). VCAM-1 was stained by addition of streptavidin-alkaline phosphatase binder (30 min, RT) and incubated with BT red (Biotec, 10, RT). An image analysis system was used to capture images of the stained tissue to determine the percentage of aortic endothelial indicative of immunoreactivity with anti-VCAM-1 antibodies.
    In addition, the in vivo activity of these compounds could be evaluated in other inflammatory models predicted to have increased VCAM-1 levels. One such model for respiratory diseases such as asthma is an ovalbumin sensitized model [Kung, T.T. Int. Arch. Allergy Immunol. 105, 83-90 (1994)]. This lung inflammation model is IgE mediated and includes eosinophilia (as in asthma patients). Bronchoalveolar lavage (BAL) fluids from laboratory animals could be evaluated for a number of variables including soluble adsorption molecule expression and leukocyte accumulation. Adsorption molecule expression could be assessed by immunohistochemistry in the tissues of experimental animals, particularly in the lungs. The effect of the claimed compounds, such as MDL 29353, should inhibit the increase in VCAM-1 expression and inhibit eosinophil accumulation in BAL fluid. Inhibitors can be tested in a rat model of adjuvant arthritis, which is reported to respond to anti-VCAM-1 monoclonal antibodies [Iigo, Y. et al. J. Immunol. 147, 4167-4171 (1991)]. In this model, adsorption expression can be assessed in the limbs (joints) of laboratory animals. For autoimmune diabetes, the ability of compounds in the NOD mouse model to delay the onset or prevent the application metastasis of the disease was tested [Heinke, E. W. et al. Diabetes 42, 1721-1730 (1993); Baron, J. L. et al. J. Clin. Invest. 93, 1700-1708 (1994). Moreover, it is possible to monitor the level of VCAM-1 expression in tissues as well as monitor the progress of diabetes in laboratory animals. The therapeutic potential for transplant rejection could be assessed by monitoring cardiac tagogy survival (Balb / c heart implanted with C3H / He receptor) [Isobe, M. et al. 153, 5810-5818 (1994). In vivo administration of anti-VCAM-1 and anti-VLA-4 single cell antibodies induces immunosuppression against cardiac grafts and soluble antigens in this mouse model. The effect of the compound on tumor metastasis and angiogenesis can be evaluated in a number of models. These may include B16 (murine) and M24met (human) melanoma models for experimental metastases [Fidler, I. J., Cancer Res. 35 218-224 (1975); Meuller, M. M. et al., Cancer Res. 51, 2193-2198. The activity of the compounds could be assessed not only by their effect on the number of advanced lung metastases, but also by the effect on VCAM-1 expression in the lung as described above for the mouse respiratory model. Models for evaluating anti-angiogenic compounds that can be used to test compounds include monitoring vascular response to a mixture of angiogenic factors mixed with basement membrane proteins injected subcutaneously in mice [Passaniti, A. et al. Lab. Invest. 67, 519-528 (1992). Angiogenesis is recorded according to the number of tubes supplemented into matrigel and the hemoglobin content of the gel. Adsorption molecule expression and leukocyte accumulation could be measured by immunohistochemistry as in all of the above examples.
    result
    The claimed compounds inhibit the cytokine-induced increase in VCAM-1 gene expression in vascular cells ex vivo. Compared to another cytokine-induced adsorption molecule, ICAM-1, selective inhibition of VCAM-1 expression was seen in certain claimed compounds (Table 3). In vivo experiments show that MDL 29353, when properly accumulated, can inhibit LPS induced levels of VCAM-1 expression in rabbit aortic endothelial (FIG. 1). This experiment also shows the fact that the compound has oral activity.
    权利要求:
    Claims (46)
    [1" claim-type="Currently amended] Administering the cytokine-induced expression of vascular cell adsorption molecule-1 to a patient in need thereof, comprising administering an inhibitory effective amount of a compound of formula (1) to vascular cell adsorption molecule-1 A method of inhibiting induced expression.
    <Formula 1>

    Where
    R 1 , R 2 , R 3 and R 4 are each independently a C 1 -C 6 alkyl group,
    Z is a thio, oxy or methylene group,
    A is a C 1 -C 4 alkylene group,
    R 5 is C 1 -C 6 alkyl or-(CH 2 ) n- (Ar), where n is an integer 0, 1, 2 or 3, and Ar is hydroxy, methoxy, ethoxy, chloro, fluoro Or phenyl or naphthyl unsubstituted or substituted with one to three substituents selected from the group consisting of C 1 -C 6 alkyl.
    [2" claim-type="Currently amended] The method of claim 1, wherein R 1 and R 2 are t-butyl.
    [3" claim-type="Currently amended] The method of claim 2, wherein R 3 and R 4 are methyl.
    [4" claim-type="Currently amended] The method of claim 3 wherein A is methylene.
    [5" claim-type="Currently amended] The method of claim 4, wherein Z is thio.
    [6" claim-type="Currently amended] The method of claim 4, wherein Z is oxy.
    [7" claim-type="Currently amended] The process of claim 1 wherein the compound is 2,6-di-t-butyl-4-[(dimethylphenylsilyl) methylthio] phenol.
    [8" claim-type="Currently amended] The process of claim 1 wherein the compound is 2,6-di-t-butyl-4-[(trimethylsilyl) methylthio] phenol.
    [9" claim-type="Currently amended] The process of claim 1 wherein the compound is 2,6-di-t-butyl-4-[(4-chlorophenyldimethylsilyl) methyloxy] phenol.
    [10" claim-type="Currently amended] The method of claim 1 wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-4-fluorophenylsilyl) methyloxy] phenol.
    [11" claim-type="Currently amended] The method of claim 1 wherein the compound is 2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol.
    [12" claim-type="Currently amended] The method of claim 1, wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-4-methoxyphenylsilyl) methyloxy] phenol.
    [13" claim-type="Currently amended] The method of claim 1 wherein the compound is 2,6-dimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol.
    [14" claim-type="Currently amended] The method of claim 1, wherein the compound is 2-t-butyl-6-methyl-4-[(dimethylphenylsilyl) methylthio] phenol.
    [15" claim-type="Currently amended] The method of claim 1 wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-2-methoxyphenylsilyl) methyloxy] phenol.
    [16" claim-type="Currently amended] The process of claim 1 wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-2,5-dimethoxyphenylsilyl) methyloxy] phenol.
    [17" claim-type="Currently amended] The process of claim 1 wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-2,3-dimethoxyphenylsilyl) methyloxy] phenol.
    [18" claim-type="Currently amended] The method of claim 1 wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-4-t-butylphenylsilyl) methyloxy] phenol.
    [19" claim-type="Currently amended] The process of claim 1 wherein the compound is 2,6-di-t-butyl-4-[(benzyldimethylsilyl) methyloxy] phenol.
    [20" claim-type="Currently amended] The process of claim 1 wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-p-methoxybenzylsilyl) methyloxy] phenol.
    [21" claim-type="Currently amended] A method of treating a patient with chronic inflammatory disease comprising administering a therapeutically effective amount of a compound of formula (1) to a patient with chronic inflammatory disease.
    <Formula 1>

    Where
    R 1 , R 2 , R 3 and R 4 are each independently a C 1 -C 6 alkyl group,
    Z is a thio, oxy or methylene group,
    A is a C 1 -C 4 alkylene group,
    R 5 is a C 1 -C 6 alkyl group or-(CH 2 ) n- (Ar), where n is an integer 0, 1, 2 or 3, and Ar is hydroxy, methoxy, ethoxy, chloro, fluoro Or phenyl or naphthyl unsubstituted or substituted with one to three substituents selected from the group consisting of C 1 -C 6 alkyl.
    [22" claim-type="Currently amended] The method of claim 21, wherein R 1 and R 2 are t-butyl.
    [23" claim-type="Currently amended] The method of claim 22, wherein R 3 and R 4 are methyl.
    [24" claim-type="Currently amended] The method of claim 23, wherein A is methylene.
    [25" claim-type="Currently amended] The method of claim 24, wherein Z is thio.
    [26" claim-type="Currently amended] The method of claim 24, wherein Z is oxy.
    [27" claim-type="Currently amended] The method of claim 21, wherein the compound is 2,6-di-t-butyl-4-[(dimethylphenylsilyl) methylthio] phenol.
    [28" claim-type="Currently amended] The method of claim 21, wherein the compound is 2,6-di-t-butyl-4-[(trimethylsilyl) methylthio] phenol.
    [29" claim-type="Currently amended] The method of claim 21, wherein the compound is 2,6-di-t-butyl-4-[(4-chlorophenyldimethylsilyl) methyloxy] phenol.
    [30" claim-type="Currently amended] The method of claim 21, wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-4-fluorophenylsilyl) methyloxy] phenol.
    [31" claim-type="Currently amended] The method of claim 21, wherein the compound is 2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol.
    [32" claim-type="Currently amended] The method of claim 21, wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-4-methoxyphenylsilyl) methyloxy] phenol.
    [33" claim-type="Currently amended] The method of claim 21, wherein the compound is 2,6-dimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol.
    [34" claim-type="Currently amended] The method of claim 21, wherein the compound is 2-t-butyl-6-methyl-4-[(dimethylphenylsilyl) methylthio] phenol.
    [35" claim-type="Currently amended] The method of claim 21, wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-2-methoxyphenylsilyl) methyloxy] phenol.
    [36" claim-type="Currently amended] The method of claim 21, wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-2,5-dimethoxyphenylsilyl) methyloxy] phenol.
    [37" claim-type="Currently amended] The method of claim 21, wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-2,3-dimethoxyphenylsilyl) methyloxy] phenol.
    [38" claim-type="Currently amended] The method of claim 21, wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-4-t-butylphenylsilyl) methyloxy] phenol.
    [39" claim-type="Currently amended] The method of claim 21, wherein the compound is 2,6-di-t-butyl-4-[(benzyldimethylsilyl) methyloxy] phenol.
    [40" claim-type="Currently amended] The method of claim 21, wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-p-methoxybenzylsilyl) methyloxy] phenol.
    [41" claim-type="Currently amended] The method of claim 21, wherein the inflammatory disease is asthma.
    [42" claim-type="Currently amended] The method of claim 21, wherein the inflammatory disease is chronic inflammation.
    [43" claim-type="Currently amended] The method of claim 21, wherein the inflammatory disease is rheumatoid arthritis.
    [44" claim-type="Currently amended] The method of claim 21, wherein the inflammatory disease is autoimmune diabetes.
    [45" claim-type="Currently amended] The method of claim 21, wherein the inflammatory disease is transplant rejection.
    [46" claim-type="Currently amended] The method of claim 21, wherein the inflammatory disease is tumor angiogenesis.
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    同族专利:
    公开号 | 公开日
    JP2008088183A|2008-04-17|
    ZA9703571B|1997-10-30|
    CA2252869A1|1997-11-06|
    NO985040D0|1998-10-29|
    WO1997040837A1|1997-11-06|
    NO985040L|1998-10-29|
    BR9709189A|1999-08-10|
    EP0910384A1|1999-04-28|
    CN1216921A|1999-05-19|
    AU2209097A|1997-11-19|
    JP2000509070A|2000-07-18|
    IL126744D0|1999-08-17|
    AR006891A1|1999-09-29|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    法律状态:
    1996-04-30|Priority to US64147496A
    1996-04-30|Priority to US8/641,474
    1997-03-12|Application filed by 게리 디. 스트리트, 스티븐 엘. 네스비트, 훽스트 마리온 로우셀, 인크.
    1997-03-12|Priority to PCT/US1997/003928
    2000-11-06|Publication of KR20000065102A
    优先权:
    申请号 | 申请日 | 专利标题
    US64147496A| true| 1996-04-30|1996-04-30|
    US8/641,474|1996-04-30|
    PCT/US1997/003928|WO1997040837A1|1996-04-30|1997-03-12|Method of inhibiting vascular cell adhesion molecule-1 and treating chronic inflammatory diseases with 2,6-di-alkyl-4-silyl-phenols|
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