Alkyl-4-silyl-phenols and their esters as atherosclerotic inhibitors

专利摘要:
The present invention is useful for treating atherosclerosis and chronic inflammatory diseases, inhibiting cytokine induced expression of VCAM-1 and / or ICAM-1, inhibiting peroxidation of LDL lipids and lowering plasma cholesterol; A compound of formula (1) or a pharmaceutically acceptable salt thereof useful as an antioxidant useful in preventing oxidative degradation in organic materials. <Formula 1> In the above formula, R 1 and R 6 are each independently C 1 -C 6 alkyl; R 2 , R 3 and R 4 are each independently hydrogen or C 1 -C 6 alkyl; R is hydrogen or —C (O) — (CH 2 ) m -Q wherein Q is hydrogen or —COOH and m is an integer of 1, 2, 3 or 4; Z is a thio, oxy or methylene group; A is a C 1 -C 4 alkylene group; R 5 and R 7 are each independently C 1 -C 6 alkyl or — (CH 2 ) n — (Ar), where n is an integer 0, 1, 2 or 3; Ar is hydroxy, methoxy, 1 to 1 selected from the group consisting of oxy, halogen, trifluoromethyl, C 1 -C 6 alkyl, or —NR 8 R 9 , wherein R 8 and R 9 are each independently hydrogen or C 1 -C 6 alkyl. Phenyl or naphthyl unsubstituted or substituted with three substituents; Provided that when at least one of R 2 and R 5 or R 7 is C 1 -C 6 alkyl and Ar is not substituted with trifluoromethyl or —NR 8 R 9 , then R is —C (O) — (CH 2 ) m -Q.
公开号:KR20000065101A
申请号:KR1019980708692
申请日:1997-03-03
公开日:2000-11-06
发明作者:스티븐 제이. 부쉬；킴 에스. 첸；마이클 제이. 에드워즈；제임스 이. 제이알. 매트；로저 에이. 파커；마크 제이. 바알；폴 에스. 라이트；마크 티. 예이츠
申请人:게리 디. 스트리트, 스티븐 엘. 네스비트；훽스트 마리온 로우셀, 인크.；
IPC主号:

专利说明:

Alkyl-4-silyl-phenols and their esters as atherosclerosis inhibitors
Coronary heart disease (CHD) is still the leading cause of death in industrialized countries. Although CHD mortality has declined in recent years, CHD still causes more than 500,000 deaths per year in the United States. The US spends more than $ 100 billion annually, directly and indirectly, due to CHD. The main cause of CHD is atherosclerosis, a disease characterized by deposition of lipids in the arterial vessel walls, narrowing the vascular pathway and eventually curing the vascular system.
Atherosclerosis, a major clinical complication of ischemic heart disease, suggests that smooth muscle cells in the artery proliferate from the erythematous layer to the arterial endocardium as lipids deposit and foam cells accumulate in the lesions after local injury begins. It is becoming. As atherosclerosis plaques develop, blood vessels may become more and more occluded gradually, eventually leading to local ischemia or infarction. Therefore, it is desirable to provide such a method of inhibition in a patient in need of inhibiting the progression of atherosclerosis.
Hypercholesterolemia is an important risk factor associated with CHD. For example, in December 1984, the National Institute of Health Consensus Development Conference Panel (NICP) found that lowering plasma cholesterol levels (especially blood levels of low-density lipoprotein cholesterol) reduced the risk of heart attack due to CHD. We conclude that it will definitely reduce. Serum lipoproteins are carriers of lipids in the circulation. They are classified according to density into taste grains, very low density lipoproteins (VLDL), low density lipoproteins (LDL), and high density lipoproteins (HDL). Grace grains are primarily involved in transporting dietary triglycerides and cholesterol from the intestine to adipose tissue and liver. VLDL carries endogenously synthesized triglycerides from the liver to fat and other tissues. LDL carries cholesterol to peripheral tissues and regulates endogenous cholesterol levels in those tissues. HDL carries cholesterol from peripheral tissues to the liver. Arterial wall cholesterol is derived almost exclusively from LDL (Brown and Goldstein, Ann. Rev. Biochem. 52, 223 (1983); Miller, Ann. Rev. Med. 31, 97 (1980)). Atherosclerosis rarely develops in patients with low LDL levels. Therefore, it would be desirable to provide a method for lowering plasma cholesterol in patients with hypercholesterolemia or at risk for developing hypercholesterolemia.
Increased cholesterol levels are also associated with numerous disease states, including stenosis, angina, cerebral atherosclerosis, and xanthoma. It would be desirable to provide a method for lowering plasma cholesterol in patients with stenosis, angina, cerebral atherosclerosis, xanthoma and other disease states associated with increased cholesterol levels or at risk of developing the disease.
Vascular Cell Adsorption Molecule-1 (VCAM-1) and Intercellular Adsorption Molecule-1 (ICAM-1) are Interleukin-1 (IL-1), Interleukin-4 (IL-4) and Tumor Necrosis in Vascular Endothelial and Smooth Muscle Cells Adsorption molecules in the class of immunoglobulins that are augmented by cytokines such as factor-α (TNF-α). Through interaction with the appropriate integrin counter receptor, VCAM-1 and ICAM-1 mediate the adsorption and endothelial metastasis of leukocytes in the inflammatory response. Inhibitors of VCAM-1 and / or ICAM-1 have therapeutic utility in various types of chronic infectious diseases, including atherosclerosis, asthma, rheumatoid arthritis and autoimmune diabetes. For example, in-situ hybridization and immunohistochemical analysis of atherosclerotic plaques from patients demonstrated that levels of adsorption molecules (VCAM-1 and ICAM-1) were increased compared to disease free sites ( O'Brien, KD et al., J. Clin. Invest. 92, 945-951 (1993); Davies, MJ et al., J. Pathol. 171, 223-229 (1993); Poston, RN et al. Am. J. Pathol. 140, 665-673 (1992). Diets that form atherosclerosis induce VCAM-1 expression in aortic endothelial and vascular smooth muscle cells in atheromatous atherosclerosis in rabbits (Poston, RN et al., Ibid .; Cybulsky, MI et al., Science 251, 788-791 ( 1991); Li, H. et al., Arterioscler. Thromb. 13, 197-204 (1993). In view of this previous study, it is believed that increased VCAM-1 expression is associated with the onset and progression of atherosclerosis plaques through supplementation to circulating mononuclear cells.
Moreover, VCAM-1 is also involved as a vehicle in other chronic inflammatory disorders such as asthma, rheumatoid arthritis and autoimmune diabetes. For example, it is known that expression of VCAM-1 and ICAM-1 is increased in asthmatic patients (Pilewski, JM et al., Am. J. Respir. Cell Mol. Biol. 12, 1-3 (1995) Ohkawara, Y. et al., Am. J. Respir. Cell Mol. Biol. 4-12 (1995). In addition, blocking the integrin receptors for VCAM-1 and ICAM-1 (VLA-4 and LFA-1, respectively) in the ovalbumin-sensitized rat model of allergic airway response inhibited both early and late responses (Rabb , HA et al., Am. J. Respir.Care Med. 149, 1186-1191 (1994)). In the microvascular structure of the rheumatoid synovial membrane, expression of endothelial adsorption molecules including VCAM-1 was also increased (Koch, AE et al., Lab. Invest. 64, 313-322 (1991); Morales-Ducret, J. et al., Immunol. 149, 1421-1431 (1992)). Neutralizing antibodies directed against VCAM-1 or its counter receptor VLA-4 can delay the onset of diabetes in a spontaneously developing mouse model (NOD mice) (Yang. XD et al., Proc. Natl. Acad Sci. USA 90, 10494-10498 (1993); Burkly, LC et al., Diabetes 43, 523-534 (1994); Baron, JL et al., J. Clin.Invest. 93, 1700-1708 (1994) )). Monoclonal antibodies against VCAM-1 may also have beneficial effects in animal models of two-factor graft rejection, suggesting that VCAM-1 expression inhibitors may have utility in preventing graft rejection (Orocz, CG). et al., Immunol. Lett. 32, 7-12 (1992).
VCAM-1 is expressed by cells as both membrane bound and soluble forms. Soluble forms of VCAM-1 have been found to induce chemotaxis of vascular endothelial cells in vivo, stimulating angiogenesis responses in rat corneas (Koch, A.E. et al., Nature 376, 517-519 (1995)). Inhibitors of expression of soluble VCAM-1 have strong therapeutic value in treating diseases with strong angiogenic components including tumor proliferation and metastasis (Folkman, J. and Shing, Y., J. Biol. Chem. 10931-). 10934 (1992).
Promoters for VCAM-1 and ICAM-1 have been cloned and characterized. For example, both promoters contain multiple DNA sequence elements capable of binding to the transcription factor NF-kB (Iademarco, MF et al., J. Biol. Chem. 267, 16323-16329 (1992); Voraberger, G. et al., J. Immunol. 147, 2777-2786 (1991)). The NF-kB family of transcription factors is central to the regulation of several genes that are extended within the site of transmission. Activation of NF-kB as a transcription factor involves separation from IkB, an inhibitory subunit in the cytoplasm. NF-kB subunits translocate to the nucleus, bind specific DNA sequence elements, and activate transcription of several genes, including VCAM-1 and ICAM-1 (Collins T. et al., Lab. Invest. 68, 499-508 (1993).
It has been hypothesized that regulation of VCAM-1 gene expression can be coupled with oxidative stress through specific reduction-oxidation (redox) sensitive transcription or post-transcriptional regulatory factors. Antioxidants pyrrolidine dithiocarbamate and N-acetylcysteine inhibit cytokine induced expression of VCAM-1 but not ICAM-1 in vascular endothelial cells (Mauri, N. et al., J. Clin. Invest 92, 1866-1874 (1993). This indicates that inhibition of VCAM-1 expression by antioxidants is associated with several additional factors that are not involved in regulating ICAM-1 expression.
2,6-di-alkyl-4-silyl-phenols are disclosed as atherosclerosis inhibitors in US Pat. No. 5,155,250, issued October 13, 1992 to Parker et al. Moreover, 2,6-di-alkyl-4-silyl-phenols are disclosed as serum cholesterol lowering agents in PCT International Publication No. WO 95/15760, published June 15, 1995.
It would be advantageous to modulate the release of VCAM-1 and / or ICAM-1 and to treat VCAM-1 and / or ICAM-1 mediated effects. It would also be advantageous to control or treat chronic inflammation without the occurrence of side effects known to accompany the use of anti-inflammatory steroids and nonsteroidal anti-inflammatory agents.
Summary of the Invention
The present invention provides a compound of formula (1) or a pharmaceutically acceptable salt thereof.
In the above formula,
R ₁ and R ₆ are each independently C ₁ -C ₆ alkyl;
R ₂ , R ₃ and R ₄ are each independently hydrogen or C ₁ -C ₆ alkyl;
R is hydrogen or —C (O) — (CH ₂ ) _m -Q wherein Q is hydrogen or —COOH and m is an integer of 1, 2, 3 or 4;
Z is a thio, oxy or methylene group;
A is a C ₁ -C ₄ alkylene group;
R ₅ and R ₇ are each independently C ₁ -C ₆ alkyl or — (CH ₂ ) _n — (Ar), where n is an integer 0, 1, 2 or 3; Ar is hydroxy, methoxy, 1 to 1 selected from the group consisting of oxy, halogen, trifluoromethyl, C ₁ -C ₆ alkyl, or —NR ₈ R ₉ , wherein R ₈ and R ₉ are each independently hydrogen or C ₁ -C ₆ alkyl. Phenyl or naphthyl unsubstituted or substituted with three substituents; Provided that when at least one of R ₂ and R ₅ or R ₇ is C ₁ -C ₆ alkyl and Ar is not substituted with trifluoromethyl or —NR ₈ R ₉ , then R is —C (O) — (CH ₂ ) _m -Q.
The present invention also provides a method for inhibiting peroxidation of LDL lipids in a patient comprising administering an effective amount of a compound antioxidant of formula (1) to a patient in need thereof.
The present invention also provides a method for lowering plasma cholesterol levels in a patient comprising administering to the patient in need of lowering plasma cholesterol levels a compound plasma cholesterol lowering amount of the formula (1).
In addition, the present invention comprises administering to a patient in need of inhibiting the progression of atherosclerosis and / or treating atherosclerosis an effective amount of a compound atherosclerosis inhibiting effective amount of the compound of formula (1) Provided are methods for inhibiting and / or treating atherosclerosis.
In addition, the present invention provides a method for administering an effective amount of the compound expression inhibiting compound of formula (1) to a patient in need of inhibiting cytokine-induced expression of vascular cell adsorption molecule-1 and / or intercellular adsorption molecule-1 It provides a method of inhibiting the expression of the molecule in the patient.
The present invention also provides a method of treating a patient, comprising administering a therapeutically effective amount of the compound of formula (1) to a patient suffering from a chronic inflammatory disease.
As used herein, the term “C ₁ -C ₆ alkyl” refers to a saturated hydrocarbyl radical of straight, branched or ring configuration consisting of 1 to 6 carbon atoms. Within the scope of this term are included methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, secondary butyl, tert-butyl, n-pentyl, n-hexyl, cyclohexyl and the like.
Likewise, the term “C ₁ -C ₄ alkylene” refers to a saturated hydrocarbyldiyl radical in a straight or branched chain arrangement of 1 to 4 carbon atoms. 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-propane-diyl, 1,4 Butane-diyl and the like.
When R ₅ is a-(CH ₂ ) _n- (Ar) radical, the "-(CH ₂ ) _n- " moiety represents a saturated hydrocarbyldiyl radical in a straight chain arrangement. The term "n" is defined as an integer 0, 1, 2 or 3. Thus, the residue "-(CH ₂ ) _n- " refers to a bond, methylene, 1,2-ethanediyl or 1,3-propanediyl. "-(Ar)" residue represents an aryl radical defined as a substituted or unsubstituted phenyl or naphthyl group. In case the-(Ar) residue is substituted aryl, phenyl or naphthyl may contain 1 to 3 substituents at any position, otherwise 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. In particular, phenyl within the scope of the term "-(CH ₂ ) _n- (Ar)";Naphthyl;Phenylmethyl;Phenylethyl;3,4,5-trihydroxyphenyl;3,4,5-trimethoxyphenyl;3,4,5-triethoxyphenyl;4-chlorophenyl;4-methylphenyl;3,5-di-tert-butyl-4-hydroxyphenyl;4-fluorophenyl;4-chloro-1-naphthyl;2-methyl-1-naphthylmethyl;2-naphthylmethyl;4-chlorophenylmethyl;4-tert-butylphenyl; 4-tert-butylphenylmethyl, and the like.
The term "pharmaceutically acceptable acid addition salt" means any of the compounds of formula (1) as appropriate, such as 2,6-di-t-butyl-4 [(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol It is intended for use in non-toxic organic or inorganic acid addition salts. Examples of inorganic acids that generate suitable salts include hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid; And acid metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Examples of organic acids that result in suitable salts include mono, di and tricarboxylic acids. Examples of such acids are acetic acid, trifluoroacetic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxide Hydroxybenzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, 2-phenoxybenzoic acid and sulfonic acid (eg, methane sulfonic acid and 2-hydroxyethane sulfonic acid). Such salts may be present in hydrated or substantially anhydrous form.
Compounds of formula (1) may be prepared using methods and techniques well known to those skilled in the art. General synthetic schemes for preparing compounds of formula (1) wherein Z is sulfur or oxygen are described in Scheme A, where all substituents are as defined above unless stated otherwise.
In general, the phenols of structure 1a may be substituted with suitable alkyl-4-mercaptophenols or alkylhydroquinones (or appropriately protected derivatives) of structure 2 such as non-prochines such as sodium hydride, potassium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide and the like. Prepared by reacting a suitable haloalkylenesilane of structure 3 such as a nuclear base and a suitable chloroalkylenesilane in a suitable protic solvent such as acetonitrile, dimethylformamide or dimethylacetamide or in an aqueous solvent such as water / 2-butanone Can be.
The phenol esters of structure 1b can be prepared by acylating the phenols of structure 1a according to standard acylation reaction techniques. For example, the phenol of structure 1a is dissolved in a suitable protic solvent such as acetonitrile, dimethylformamide or dimethylacetamide or an ether solvent such as diethyl ether or dioxane, triethylamine, N-methylmorpholine Treatment with a suitable base such as sodium hydroxide or sodium hydride. An excess of O-acylating agent is then added at room temperature and the reaction mixture is stirred at room temperature for 1 to 24 hours. Examples of O-acylating agents are acetyl chloride, propionyl chloride, monoethylsuccinyl chloride, succinic anhydride and the like. The product is purified by techniques well known in the art, such as extraction and flash chromatography. Optionally, further treatment with a suitable base such as sodium hydroxide, acidified with a suitable acid such as hydrochloric acid, followed by extraction and flash chromatography can yield the phenol ester of structure 1b.
The starting materials used in the general synthesis outlined in Scheme A are readily available to those of ordinary skill in the art. Certain phenolic starts of various compounds of formula (1) wherein Z is sulfur, such as, for example, 2,6-di-tert-butyl-4-mercaptophenol and 2-3-butyl-4-mercaptophenol Materials are disclosed in U.S. Patent Nos. 3,576,883, 3,952,064, 3,479407, 4,975,467, 5,155,250, and Japanese Patent Application 73-28425. Other phenol starting materials for the compound of formula (1) include commercially available trimethylhydroquinone and 2,5-di-tert-butylhydroquinone.
Silyl starting materials for various compounds of formula (1) such as (trimethylsilyl) methyl iodide, (trimethylsilyl) methyl bromide, (trimethylsilyl) methyl chloride and (1-chloropropyl) trimethylsilane are described in Synthesis 4 , 318-19 (1988) and J. Am. Chem. Soc. 105, 5665-75 (1983).
Additional methods of preparing suitable silanes include the Grignard reaction. For example, when R ₇ is a phenyl moiety containing a methoxy substituent, 4-bromoanisole is reacted with magnesium metal to produce a Grignard reagent, which is reacted with chlorodimethyl chloromethyl silane to Obtain methyldimethyl-4-methoxy phenyl silane.

Alternatively, anisole is reacted with n-butyllithium to lithiate and the resulting thiothio compound is reacted with chlorodimethyl chloromethyl silane to give chloromethyl dimethyl-2-methoxyphenyl silane.

When R ₇ is a phenyl moiety containing an -NR ₈ R ₉ substituent such as dimethylamine, 4-bromo-N, N-dimethylaniline or the like is reacted with magnesium metal to generate a Grignard reagent, Reaction with chlorodimethyl chloromethyl silane yields 4-N, N-dimethylaminophenyl (dimethyl) chloromethylsilane.
When R ₇ is a phenyl moiety containing a trifluoromethyl substituent, bromo benzotrifluoride is reacted with magnesium metal to generate a Grignard reagent, which is reacted with chlorodimethyl chloromethyl silane to give dimethyl (chloro Methyl) trifluoromethylphenylsilane is obtained.
When both R ₅ and R ₇ are phenyl, about 2 molar equivalents of phenylmagnesium bromide are reacted with about 1 molar equivalent of dichloromethyl chloromethyl silane to obtain methyl diphenylchloromethyl silane.
In the case of 1-phenol functional groups, compounds of structure 2 can be reacted with compounds of structure 3 under reaction conditions, and the 1-phenol functions of structure 2 can be blocked with standard phenol blockers well known in the art. The selection and use of specific circuit breakers are well known to those skilled in the art. In general, blockers should be appropriately selected to adequately protect the phenols during subsequent synthesis steps and should be readily removable under conditions that do not cause degradation of the desired product.
Examples of suitable phenol protecting groups include ethers such as methoxymethyl, 2-methoxyethoxymethyl, tetrahydropyranyl, t-butyl and benzyl ethers; Silyl ethers such as trimethylsilyl and t-butyldimethylsilyl ether; Esters such as acetates and benzoates; Carbonates such as methyl carbonate and benzyl carbonate; Sulfonates such as methanesulfonate and toluenesulfonate.
In the case where R ₁ and R ₂ are each t-butyl, the reaction of Scheme A may be conveniently performed without blocking the 1-phenol functional group.
The following examples show representative synthesis examples described in Scheme A. Such embodiments are to be understood as illustrative only and are not intended to limit the scope of the invention in any way. As used herein, the term "g" refers to grams; "mol" refers to mole; "mmol" refers to millimoles; "L" refers to liters; "Ml" refers to milliliters; "° C" refers to degrees Celsius; "mm Hg" refers to millimeters of barometer mercury; "Mg" refers to milligrams; "μM" refers to micro molarity; "Μg" refers to micrograms.
<Example 1>
2,6-di-t-butyl-4-[(diphenylmethylsilyl) methyloxy] phenol (MDL 104,599)
Step a; Preparation of Chloro (diphenyl) methylsilane: 31.5 mL (0.3 mol) of bromobenzene solution was cooled to -60 ° C in 450 mL of dry THF. To this solution was added dropwise 120 ml (0.3 mol) of 2.5 M n-butyllithium while maintaining the reaction temperature below -55 ° C. When the dropwise addition was completed, 18.9 mL (0.15 mol) of chloromethyl (dichloro) methylsilane was added at a rate such that the reaction temperature was maintained below -55 ° C. The mixture was warmed to room temperature and 5 ml of ethyl acetate was added to quench any unreacted n-butyllithium. The reaction mixture was poured into 250 mL of water and the organic phase was separated. The organic phase was washed with 3 x 100 mL of water, treated with 3 x 100 mL of saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate, filtered and evaporated. The resulting pale yellow liquid was distilled at 155 to 160 ° C. and 5 mm Hg to afford the title compound as 33.1 g (90% yield) of a colorless clear liquid. GC / MS confirmed the structure and purity (about 99%) of the product.
Step b; Preparation of Diphenyl (iodomethyl) methylsilane: A solution of 20.0 g (81 mmol) of chloromethyl (diphenyl) methylsilane and 12.3 g (82 mmol) of sodium iodide in 250 mL of 2-butanone was refluxed overnight. After that, the solution was filtered and evaporated. The resulting yellow oil was redissolved in 250 ml of ethyl acetate, washed with 3 × 100 ml of water and 3 × 100 ml of saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate, filtered and evaporated. The pale yellow oil appeared as the title compound in purity (about 99%) sufficient to be used as such by GC / MS.
Step c; Preparation of 2,6-di-t-butyl-4-[(diphenylmethylsilyl) methyloxy] phenol (MDL 104,599): 9.17 g (27.1 mmol) of diphenyl (iodomethyl) methylsilane in 250 ml of anhydrous acetonitrile. ) And 6.0 g (27 mmol) of 2,6-di-t-butylbenzhydroquinone were completely degassed with nitrogen. To this solution was added 4.5 g (32.6 mmol) of potassium carbonate and the mixture was refluxed under nitrogen for 3 days. The reaction mixture was cooled down, filtered and evaporated. The resulting oil was redissolved in 250 ml of ethyl acetate, washed with 3 × 100 ml of water and 3 × 100 ml of saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate and evaporated. The oil was distilled at 250 ° C. at 5 mmHg to remove low boiling impurities, then carefully chromatographed (silica gel) several times, eluting with hexane, and finally purified by recrystallization from methanol to give the title compound at a melting point of 92 to 94 ° C. Obtained as 1.2 g (10% yield) of a white solid.
Elemental analysis for C ₂₈ H ₃₆ O ₂ Si: Calcd: C, 77.73; H, 8.39; Found: C, 77.66; H, 8.57.
NMR (CDCl ₃ ): 7.65-7.60 (m, 4H), 7.44-7.33 (m, 6H), 6.83 (s, 2H), 4.74 (s, 1H), 4.02 (s, 2H), 1.42 (s, 18H ), 0.70 (s, 3H).
<Example 2>
2,6-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol (MDL 104,556)
Step a; Preparation of Chloromethyl (dimethyl) -4-N, N-dimethylaminophenylsilane: 9.7 g (0.4 g atom) of magnesium turning was stirred under nitrogen with a Teflon paddle overnight. This "activated" magnesium was suspended in 100 ml of dry THF and iodine crystals were added. To this suspension was added a solution of 80.0 g (0.4 mol) of 4-bromo-N, N-dimethylaniline in 400 mL THF at a rate such that gentle reflux was maintained. Stirring was continued until the addition was complete (almost 2 hours) until the magnesium was consumed. A solution of 52.7 mL (0.4 mol) of chloro (chloromethyl) dimethylsilane in 220 mL of dry THF was then added dropwise and the mixture was stirred at rt overnight. The reaction mixture was quenched with 500 mL saturated aqueous ammonium chloride and stirred at room temperature (about 2 hours). The precipitated magnesium salt was filtered off and the reaction mixture was diluted with 300 mL of ether. The organic phase was separated, washed with 3 x 250 ml of water and 3 x 250 ml of saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate, filtered and evaporated. Approximately 90 g of the resulting brown oil was purified by distillation to give the title compound as 83.5 g of a colorless transparent liquid (yield 92%, boiling point 145 ° C. at 5 mmHg). GC / MS confirmed the structure and purity (about 100%) of the product.
Step b; Preparation of 4-N, N-dimethylaminophenyl (dimethyl) iodomethylsilane: 50.0 g (0.22 mol) of chloromethyl (dimethyl) -4-N, N-dimethylaminophenylsilane in 500 mL 2-butanone and iodide A solution of 33.0 g (0.22 mol) of sodium was refluxed overnight. This solution was filtered and evaporated. The resulting liquid was redissolved in 500 mL of ethyl acetate, washed with 3 × 200 mL of water and 3 × 200 mL of saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate, filtered and evaporated. The resulting pale yellow liquid was distilled at 165 ° C. and 5 mm Hg to give the title compound as 63.7 g (91% yield) of a colorless transparent liquid. GC / MS confirmed the structure and purity (about 100%) of the product.
Step c; Preparation of 2,6-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol (MDL 104,556): 4-N, N-dimethyl in 1.5 L of anhydrous acetonitrile A solution of 444 g (1.39 mol) of aminophenyl (dimethyl) iodomethylsilane and 309 g (1.39 mol) of 2,6-di-t-butylbenzhydroquinone was completely degassed with nitrogen. To this solution was added 435 g (1.39 mol) cesium carbonate and the mixture was refluxed under nitrogen for 24 hours. The reaction mixture was cooled, diluted with 1.5 L of ethyl acetate, washed with 3 × 500 mL of water and 3 × 500 mL of saturated sodium chloride, dried over anhydrous magnesium sulfate and evaporated. The resulting brown solid was treated with 1 L of methanol, filtered and dried under vacuum to give 350 g of a rose solid. This material was recrystallized from ethyl acetate / methanol (about 1: 10, 2 L). The material provided as a waxy white solid was homogenized by mechanical stirring with a Teflon paddle. This waxy solid was filtered, washed with 1 L of cooled methanol in dry ice / acetone and dried under vacuum to afford the title compound as 260 g (yield 45%) of a white solid with a melting point of 115 to 177 ° C.
Elemental analysis for C ₂₅ H ₃₉ NO ₂ Si: Calcd: C, 72.59; H, 9. 50; N, 3.39; Found: C, 72.47; H, 9. 50; N, 3.32.
NMR (CDCl ₃ ): 7.47 (d, 2H, J = 8.6 Hz), 6.81 (s, 2H), 6.75 (d, 2H, J = 8.6 Hz), 4.70 (s, 1H), 3.69 (s, 2H) , 2.96 (s, 6H), 1.42 (s, 18H), 0.38 (s, 6H).
<Example 3>
2,6-di-t-butyl-4-[(dimethyl-4-trifluoromethylphenyldimethylsilyl) methyloxy] phenol (MDL 105,975)
Step a; Preparation of Dimethyl-4-trifluoromethylphenylchloromethylsilane: 9.7 g (0.4 mol) of magnesium metal shaving is placed in a three-neck flask and the metal is stirred overnight with an overhead stirrer under nitrogen atmosphere. Activated. 100 mL of THF and iodine crystals were added and a solution of 90 g (0.4 mol) of 4-bromobenzotrifluoride in 500 mL of THF was added at a rate that maintained reflux. The mixture was further stirred for 4 hours, a solution of 57.2 g (0.4 mol) of chlorodimethylchloromethylsilane in 100 ml of THF was added at a rate that remained close to reflux and the mixture was stirred overnight at ambient atmosphere. This mixture was poured into a mixture of ether / aqueous ammonium chloride (1 L each), the organic layer was isolated, dried and evaporated. The residue was distilled off to give 53 g (53%) of the title compound as a clear liquid with a boiling point of 87-89 ° C. at 0.1 mmHg.
Elemental Analysis for C ₁₀ H ₁₂ ClF ₃ Si: Calcd: C, 47.52; H, 4.79; Found: C, 47.31; H, 4.77.
Step b; Preparation of 2,6-di-t-butyl-4-[(dimethyl-4-trifluoromethylphenylsilyl) methyloxy] phenol (MDL 105,975): Dimethyl (iodomethyl) -4- in 200 mL of anhydrous acetonitrile A solution of 12.0 g (35 mmol) of trifluoromethylphenylsilane and 6.5 g (29.2 mmol) of 2,6-di-t-butylbenzhydroquinone was completely degassed with nitrogen. To this solution was added 4.8 g (35 mmol) of potassium carbonate and the mixture was refluxed under nitrogen for 36 hours. The reaction mixture was cooled down, filtered and evaporated. The resulting oil was redissolved in 250 ml of ethyl acetate, washed with 3 × 100 ml of water and 3 × 100 ml of saturated sodium chloride, dried over anhydrous magnesium sulfate and evaporated. The oil was purified by distillation at 250 ° C. at 5 mmHg to remove low boiling impurities and the product was distilled (boiling points 215-220 ° C. at 5 mM Hg). 6.87 g of the title compound, which crystallizes when left, was recrystallized from methanol and dried under vacuum to give 3.95 g (31%) of a white solid having a melting point of 107 to 110 ° C.
Elemental analysis for C ₂₄ H ₃₃ F ₃ O ₂ Si: Calcd: C, 65.72; H, 7. 58; Found: C, 65.46; H, 7.46.
NMR (CDCl ₃ ): 7.73 (d, 2H, J = 7.5 Hz), 7.61 (d, 2H, J = 7.5 Hz), 6.79 (s, 2H), 4.74 (s, 1H), 3.75 (s, 2H) , 1.42 (s, 18 H), 0.44 (s, 6 H).
<Example 4>
2,6-di-t-butyl-4-[(dimethyl-3-trifluoromethylphenylsilyl) methyloxy] phenol (MDL 105,726)
Step a; Preparation of Chloromethyl (dimethyl) -3-trifluoromethylphenylsilane: 9.7 g (0.4 g atom) of magnesium turning was stirred under nitrogen overnight with a trademark Teflon paddle. This "activated" magnesium was suspended in 100 ml of dry THF and iodine crystals were added. To this suspension was added a solution of 56 mL (0.4 mol) of 3-bromo-benzotrifluoride in 400 mL of THF at a rate such that gentle reflux was maintained. Stirring was continued until the addition was complete (almost 2 hours) until the magnesium was consumed. A solution of 52.7 mL (0.4 mol) of chloro (chloromethyl) dimethylsilane in 220 mL of dry THF was then added dropwise and the mixture was stirred at rt overnight. The reaction mixture was quenched with 500 mL saturated aqueous ammonium chloride and stirred at room temperature (about 2 hours). The precipitated magnesium salt was filtered off and the reaction mixture was diluted with 300 mL of ether. The organic phase was separated, washed with 3 x 250 ml of water and 3 x 250 ml of saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate, filtered and evaporated. About 90 g of the resulting brown oil was purified by distillation to give 69.2 g of a colorless transparent liquid (yield 69%, boiling point 95 ° C. at 5 mmHg). GC / MS confirmed the structure and purity (about 100%) of the product.
Step b; Preparation of Dimethyl (iodomethyl) -3-trifluoromethylphenylsilane: 25.3 g (0.1 mol) of chloromethyl (dimethyl) -3-trifluoromethylphenylsilane in 400 mL of 2-butanone and 15.3 g (0.102) of sodium iodide mol) solution was refluxed overnight. This solution was filtered and evaporated. The resulting liquid was redissolved in 500 ml of ethyl acetate, washed with 3 × 250 ml of water and 3 × 250 ml of saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate, filtered and evaporated. The title compound produced in pale orange was pure enough (about 96%) to be used as is.
Step c; Preparation of 2,6-di-t-butyl-4-[(dimethyl-3-trifluoromethylphenylsilyl) methyloxy] phenol (MDL 105,726): Dimethyl (iodomethyl) -3- in 250 mL of anhydrous acetonitrile-3- A solution of 15.5 g (45 mmol) of trifluoromethylphenylsilane and 10 g (45 mmol) of 2,6-di-t-butylbenzhydroquinone was completely degassed with nitrogen. To this solution was added 6.2 g (45 mmol) of potassium carbonate and the mixture was refluxed under nitrogen for 3 days. The reaction mixture was cooled down, filtered and evaporated. The red oil obtained was redissolved in 250 ml of ethyl acetate, washed with 3 x 100 ml of water and 3 x 100 ml of saturated sodium chloride, dried over anhydrous magnesium sulfate and evaporated. About 24 g of the resulting red oil was distilled at 5 ° C. to 150 ° C. to remove low boiling impurities. Flash chromatography (20% CH ₂ Cl ₂ -hexane), about 12 g of material remaining in the pot, recrystallize twice from methanol and dry in vacuo to give the title compound 1.5 as a white solid at melting point 79-83 ° C. Obtained as g (yield 8%).
Elemental analysis for C ₂₄ H ₃₃ F ₃ O ₂ Si: Calcd: C, 65.72; H, 7. 58; Found: C, 65.62; H, 7.53.
NMR (CDCl ₃ ): 7.88 (m, 1H), 7.81 (dm, 1H, J = 7.3 Hz), 7.64 (dm, 1H, J = 7.3 Hz), 7.49 (t, 1H, J = 7.3 Hz), 6.81 (s, 2H), 4.75 (s, 1H), 3.76 (s, 2H), 1.44 (s, 18H), 0.46 (s, 6H).
Example 5
2-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol (MDL 103,491)
2-t-butyl-1,4-hydroquino33.2 g (0.2 mol), chloromethyldimethylphenylsilane 37.0 g (0.2 mol), lithium bromide 17.4 g (0.2 mol), potassium carbonate 27.6 g (0.2 mol) and aceto A mixture of 800 mL of nitrile was heated to reflux with stirring for 5 days. The mixture was cooled down, diluted with water and extracted with ether. The ether layer was washed with water and evaporated to dryness to give 66.1 g of dark oil. This oil was distilled in kugelrohr. Obtain 29.9 g of oil from the collected (135-155 ° C. at 0.1 mmHg) fraction, re-distillate it (135-155 ° C. at 0.1 mmHg) and chromatograph (chloroform) on silica gel to give 2-t-butyl-4- [ 29.2 g of (dimethylphenylsilyl) methyloxy] phenol were obtained as a bright yellow oil having a boiling point of 135 ° C. (0.1 mmHg).
Elemental Analysis for C ₁₉ H ₂₆ O ₂ Si: Calcd: C, 72.56; H, 8.33; Found: C, 72.32; H, 8.32.
<Example 6>
2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol succinic acid ester (MDL 103,076)
5.0 g (13.5 mmol) of 2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol (US Pat. No. 5,155,250) in 100 ml of dimethylacetamide and sodium hydride (60% in oil) 0.6 g (15 mmol) was stirred at rt for 1 h. 2.46 g (15 mmol) of monoethylsuccinylchloride were added to the reaction mixture with stirring. The reaction mixture was stirred at rt overnight, then heated at 90 ° C. for 2 h and cooled. The mixture was diluted with water and extracted with ether. The ether layer was washed with water and evaporated to dryness to give 6.6 g of a yellow oil. This oil was combined with 100 ml of methanol and heated to reflux. Sodium hydride (1.0 g in 20 mL of water) was added and the reaction mixture was refluxed for 30 minutes, then diluted with water and cooled. This aqueous suspension was acidified with concentrated hydrochloric acid and the mixture was extracted with ether and tetrahydrofuran. The organic layer was separated and evaporated to dryness to afford a yellow oil which crystallized from hexanes. 3.9 g of 2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol succinic acid ester of white crystalline powder having a melting point of 115 to 117 ° C were obtained.
Elemental Analysis for C ₂₇ H ₃₈ O ₅ Si: Calcd: C, 68.90; H, 8.14; Found: C, 68.78; H, 7.93.
<Example 7>
2-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol succinic acid ester (MDL 104,399)
2-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol (Example 5) 6.3 g (20 mmol), succinic anhydride 2.2 g (22 mmol), triethylamine 2.23 g (22 mmol) and aceto 100 ml of nitrile were combined and stirred at rt overnight, then heated to reflux for 2 h. The cooled mixture was diluted with water and extracted with ether. The ether layer was evaporated to dryness to give a white solid, which was recrystallized from acetonitrile. 6.1 g of white solids having a melting point of 92 to 93 DEG C as 2-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol succinic acid ester were obtained.
Elemental analysis for C ₂₃ H ₃₀ O ₅ Si: Calcd: C, 66.63; H, 7. 29; Found: C, 66.63; H, 7.35.
<Example 8>
2,6-di-t-butyl-4-[(dimethylphenylsilyl) methylthio] phenol succinic acid ester (MDL 103,141)
10.0 g (25.9 mmol) and 2,6-di-t-butyl-4-[(dimethylphenylsilyl) methylthio] phenol (US Pat. No. 5,155,250) in 200 mL tetrahydrofuran and sodium hydride (60% in oil) 1.03 g (25.9 mmol) of the mixture was stirred at rt for 1 h. 4.26 g (25.9 mmol) of monoethylsuccinylchloride were added to the reaction mixture with stirring. The reaction mixture was stirred at rt overnight, then heated to reflux for 2 h and cooled. The mixture was diluted with water and extracted with ether. The ether layer was washed with water and evaporated to dryness to give 12.3 g of a waxy solid. This solid was combined with 200 ml of methanol and heated to reflux. Sodium hydroxide (5.0 g in 20 ml of water) was added and the reaction mixture was refluxed for 30 minutes, then diluted with water and cooled. The aqueous suspension was acidified with concentrated hydrochloric acid and 10.6 g of solid was recovered and recrystallized from hexane to give 2,6-di-t-butyl-4-[(dimethylphenylsilyl) methylthio] at a melting point of 146 to 147 ° C. 9.3 g of a white crystalline powder that is a phenol succinic acid ester were obtained.
Elemental Analysis for C ₂₇ H ₃₈ O ₄ SSi: Calcd: C, 66.62; H, 7.87; Found: C, 66.53; H, 7.68.
Example 9
2,6-di-t-butyl-4-[(trimethylsilyl) methylthio] phenol succinic acid (MDL 104,863)
4.3 g (13.2 mmol) of 2,6-di-t-butyl-4-[(trimethylsilyl) methylthio] phenol (US Pat. No. 5,155,250) in 50 ml of dimethylacetamide and sodium hydride (60% in oil) 0.48 A mixture of g (12 mmol) was stirred at rt. 2.2 g (13.2 mmol) of monoethylsuccinylchloride were added and the mixture was stirred at rt for 3 h. The mixture was diluted with water and extracted with ether. The ether layer was washed with water and evaporated to dryness to give 5.4 g of a brown oil. The oil was chromatographed (chloroform) on silica gel to give 3.4 g of oil. This oil was combined with 50 ml of methanol and heated to reflux. Sodium hydroxide (0.6 g in 10 ml of water) was added and the reaction mixture was refluxed for 30 minutes, then diluted with water and cooled. This aqueous suspension was acidified with concentrated hydrochloric acid and extracted with ether. The ether layer was separated and evaporated to dryness to yield 3.0 g of white solid foam form. This solid was chromatographed on silica gel and then recrystallized from ethanol water to give a white crystalline powder 10.6 which is 2,6-di-t-butyl-4-[(trimethylsilyl) methylthio] phenol succinic acid ester at a melting point of 127 to 128 ° C. g was obtained.
Elemental Analysis for C ₂₂ H ₃₆ O ₄ SSi: Calcd: C, 62.22; H, 8.55; Found: C, 62.33; H, 8.69.
<Example 10>
2-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol acetic acid ester (MDL 105,443)
4.8 g (15.3 mmol), triethylamine 3.04 g (30 mmol) and 100 ml of ether were combined and stirred at room temperature in 2-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol (Example 5). . 2.4 g (30 mmol) of acetyl chloride was added slowly with stirring. The mixture was stirred for 4 hours and then diluted with water. The layers were separated and the organic layer was evaporated to dryness to give 5.6 g of oil. This oil was distilled at 150-160 ° C. (0.1 mmHg) in Kugelloh to give 5.2 g of the title compound as a light yellow oil.
Elemental Analysis for C ₂₁ H ₂₈ O ₃ Si: Calcd: C, 70.74; H, 7.92; Found: C, 71.00; H, 8.09.
<Example 11>
2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol acetic acid ester (MDL 103,377)
6.2 g (16.7 mmol) of 2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol (US Pat. No. 5,155,250), sodium hydride (60% in oil) 0.67 g (16.7 mmol) And 50 ml of dimethylacetamide were combined and stirred at room temperature for 30 minutes. 2.6 g (33.5 mmol) of acetyl chloride were slowly added to the reaction mixture and the reaction mixture was stirred overnight. The reaction mixture was diluted with water and ether and the layers separated. The ether layer was evaporated to dryness to afford 7.0 g of a waxy solid. Distilled in Kugelroer (150-165 ° C., 0.1 mmHg), and then recrystallized from hexane to give 2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol succinic acid ester acetic acid ester To a white crystalline solid at -101 占 폚.
Elemental analysis for C ₂₅ H ₃₆ O ₃ Si: Calcd: C, 72.76; H, 8.79; Found: C, 72.90; H, 8.59.
<Example 12>
2,3,6-trimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol acetic acid ester (MDL 103,157)
Step a; Preparation of 4-acetoxy-2,3,5-trimethylphenol: 15.2 g (0.1 mol) of trimethylhydroquinone, 25.3 g (0.25 mol) of triethylamine and 500 ml of ether were stirred in an ice bath. 19.6 g (0.25 mol) of acetyl chloride were slowly added with stirring, the reaction mixture was warmed for 1 hour, then diluted with water and the layers separated. The ether layer was evaporated to dryness to give 23.1 g of a tan crystalline solid diacetate having a melting point of 105 to 108 ° C. This diacetate was dissolved in 300 ml of methanol. 11 ml strong ammonium hydroxide was added and the mixture was stirred at rt overnight. The solvent was distilled off under reduced pressure and the residue was dissolved in ether. The ether layer was washed with water and evaporated to dryness to give 18.4 g of a tan solid. Recrystallization from hexane-ether gave 16.7 g of 4-acetoxy-2,3,5-trimethylphenol having a melting point of 106 to 107 캜.
Step b; Preparation of 2,3,6-trimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol acetic acid ester (MDL 103,157): 4-acetoxy-2,3,5-trimethylphenol 8.1 g (41.7 mmol), chloro 7.7 g (41.7 mmol) of methyldimethylphenylsilane, 3.6 g (41.7 mmol) of lithium bromide, 5.8 g (41,7 mmol) of potassium carbonate and 150 ml of acetonitrile were combined and heated to reflux for 3 days with stirring. The mixture was cooled down, diluted with water, acidified with concentrated hydrochloric acid and extracted with ether. The ether layer was evaporated to dryness to give 14.9 g of yellow oil. Distilled in cougelor (145-160 ° C., 0.1 mmHg) and then chromatographed (chloroform) on silica gel to give 8.6 g of the title compound as a colorless oil.
Elemental Analysis for C ₂₀ H ₂₆ O ₃ Si: Calcd: C, 71.13; H, 7.65; Found: C, 70.82; H, 7.74.
Example 13
2,5-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol (MDL 104,962)
2,5-di-t-butyl-1,4-hydroquinone (Aldrich Chemical Company, Milwaukee, WI 53233) 66.7 g (0.3 mol), chloromethyldimethylphenylsilane 55.4 g (0.3 mol), A mixture of 8.7 g (0.1 mol) of lithium bromide, 41.5 g (0.3 mol) of potassium carbonate, 2.0 g of sodium iodide and 600 ml of acetonitrile was heated to reflux for 3 days with stirring. The mixture was cooled down, diluted with water and extracted with ether. The ether layer was washed with water and evaporated to dryness to give 120 g of dark oil. This oil was distilled in cugelrore. 20.1 g of oil were obtained from the collected (150-170 ° C., 0.1 mmHg) fraction, which was chromatographed (chloroform) on silica gel to give 18.3 g of a light yellow oil.
Elemental Analysis for C ₂₃ H ₃₄ O ₂ Si: Calcd: C, 74.54; H, 9. 25; Found: C, 74.71; H, 9.27.
<Example 14>
2,5-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol acetic acid ester (MDL 106,290)
A mixture of 7.4 g (20 mmol) of triethylamine and 2.53 g (25 mmol) of 2,5-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol (Example 13) in 150 mL ether Stir at room temperature. 1.96 g (25 mmol) of acetyl chloride were added and the mixture was stirred overnight. Water and ether were added and the layers separated. The organic layer was evaporated to give 8.2 g of amber oil, which was distilled to 150-180 ° C. (0.1 mmHg) in Kugellor. Chromatography (chloroform) on silica gel gave 7.5 g of the title compound as a colorless oil.
Elemental analysis for C ₂₅ H ₃₆ O ₃ Si: Calcd: C, 72.76; H, 8.79; Found: C, 72.99; H, 8.85.
<Example 15>
2-t-butyl-4-[(dimethylphenylsilyl) methylthio] phenol (MDL 104,571)
9.1 g (50 mmol) of 2-t-butyl-4-mercaptophenol, 9.3 g (50 mmol) of chloromethyldimethylphenylsilane, 5.0 g (50 mmol) of potassium bicarbonate, 0.1 g of potassium carbonate, 2.0 g of potassium iodide and isopropanol 150 mL of the mixture was heated to reflux with stirring overnight. The mixture was cooled down, diluted with water and ether and the layers separated. The organic layer was evaporated to dryness to afford 18.0 g of amber oil, which was distilled at 150-170 ° C. in Kugelrore. Chromatography (chloroform) on silica gel gave 11.3 g of a light colorless oil.
Elemental Analysis for C ₁₉ H ₂₆ OSSi: Calcd: C, 69.03; H, 7.93; Found: C, 69.44; H, 8.05.
<Example 16>
2,3,6-trimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol (MDL 105,314)
A mixture of 10.0 g (66 mmol) of trimethylhydroquinone (Aldrich Chemical Company), 12.2 g (66 mmol) of chloromethyldimethylphenylsilane, 9.12 g (66 mmol) of potassium carbonate, 9.9 g of sodium iodide and 150 ml of acetonitrile Heated to reflux for 5 days with stirring. The mixture was cooled down, diluted with water and ether and the layers separated. The organic layer was evaporated to dryness to afford 16.2 g of amber oil, which was distilled at 145 to 165 ° C. (0.1 mmHg) in Kugellower. The resulting oil was chromatographed on chloroform (chloroform: carbon tetrachloride 1: 1) to obtain an oil, which was distilled at 145 to 155 캜 (0.1 mmHg) to give 2,3,6-trimethyl-4-[(dimethylphenylsilyl) methyloxy ] 6.2 g of phenol were obtained as light yellow.
Elemental Analysis for C ₁₈ H ₂₄ O ₂ Si: Calcd: C, 71.95; H, 8.05; Found: C, 71.88; H, 8.14.
<Example 17>
2,3,5-trimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol (MDL 103,653)
The reaction product of Example 16 was chromatographed (chloroform) and then distilled at 140-150 ° C. (0.1 mmHg) to give 0.8 g of 2,3,5-trimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol. Obtained as a colorless oil.
Elemental Analysis for C ₁₈ H ₂₄ O ₂ Si: Calcd: C, 71.95; H, 8.05; Found: C, 71.67; H, 8.08.
Example 18
2-t-butyl-4-[(dimethyl-p-methoxyphenylsilyl) methyloxy] phenol (MDL 106,834)
31.26 g (102 mmol) of iodomethyldimethyl (4-methoxy) phenylsilane, 36.6 g (112.2 mmol) of cesium carbonate and 18.6 g (112.2 mmol) of t-butylhydroquinone (Aldrich Chemical Company) in 250 ml of acetonitrile ) Was added. This mixture was heated to 90 ° C and stirred for 42 hours. The mixture was cooled to rt and filtered. Acetonitrile was removed under vacuum and the residue was dissolved in 500 ml of ethyl acetate. The organic phase was washed with 200 mL of water, dried (MgSO ₄ ) and concentrated in vacuo. The residue was purified by column chromatography three times on silica gel eluting with 20: 1 hexanes / ethyl acetate to afford the title compound as 6.8 g of a light brown oil (yield 19.3%).
Elemental analysis for C ₂₀ H ₂₈ O ₃ Si: Calcd: C, 69.72; H, 8. 19; Found: C, 69.29; H, 8.13.
Example 19
2,6-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol propionic acid ester
8.26 g (20 mmol) of triethylamine and 2,6-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol in 150 mL ether (Example 2) A mixture of g (25 mmol) was stirred at rt. 23 g (25 mmol) propionyl chloride was added and the mixture was stirred overnight. Water and ether were added and the layers separated. The organic layer was evaporated to give an oil which was distilled in kugellore. Chromatography (chloroform) on silica gel gave the title compound.
Example 20
2,6-di-t-butyl-4-[(dimethyl-4-trifluoromethylphenylsilyl) methyloxy] phenol butyric acid ester
8.76 g (20 mmol) and 2.53 g of triethylamine (2,6-di-t-butyl-4-[(dimethyl-4-trifluoromethylphenylsilyl) methyloxy] phenol in 150 ml of ether (Example 3) 25 mmol) was stirred at room temperature. 2.66 g (25 mmol) of butyryl chloride were added and the mixture was stirred overnight. Water and ether were added and the layers separated. The organic layer was evaporated to give an oil which was distilled in kugellore. Chromatography (chloroform) on silica gel gave the title compound.
Example 21
2,5-di-t-butyl-4-[(diphenylmethylsilyl) methyloxy] phenol
A solution of 9.17 g (27.1 mmol) of diphenyl (methyl) iodomethylsilane (step a of Example 1) and 6.0 g (27 mmol) of 2,5-di-t-butylbenzhydroquinone in 250 ml of anhydrous acetonitrile The gas was removed completely with nitrogen and 4.5 g (32.6 mmol) of potassium carbonate were added in a similar manner to that described in step b of Example 1 to afford the title compound.
<Example 22>
2-t-butyl-4-[(diphenylmethylsilyl) methyloxy] phenol (MDL 107,917)
A mixture of 30 g (0.18 mol) of 2-t-butylbenzhydroquinone, 45 g (0.18 mol) of methyldiphenylchloromethylsilane, 58 g (0.18 mol) of cesium carbonate and 5 g of lithium bromide in 500 ml of acetonitrile under nitrogen Heated at reflux for 7 days, cooled and poured into 1 L of water. The organic layer was isolated, dried and evaporated. The residue was placed on a Kugellor apparatus and heated at a temperature of 90 ° C. (0.1 mmHg) for 2 hours. The residue was chromatographed (hexane / ethyl acetate 9/1). The purified material was recrystallized with 0.08 mol obtained in the second run to give 16.2 g (12%) of the product as a white solid with a melting point of 100 to 101.5 ° C.
Elemental analysis for C ₂₄ H ₂₈ O ₂ Si: Calcd: C, 76.55; H, 7. 49; Found: C, 76.35; H, 7.49.
<Example 23>
2,6-di-t-butyl-4-[(methyl-di-p-methoxyphenylsilyl) methyloxy] phenol (MDL 108,208)
Step a; Preparation of Chloromethylbis (4-methoxyphenyl) methylsilane: A solution of 50 ml (0.4 mol) of 4-bromoanisole in 500 ml of THF was activating magnesium 9.7 in 100 ml of dry THF containing iodine crystals. g (0.4 g atom) was added to the suspension. A solution of 25.5 mL (0.2 mol) of chloromethyl (dichloro) methylsilane in 100 mL of dry THF was then added according to the method of step a of Example 2 to give a pale yellow oil. This pale yellow oil was distilled at 200 ° C. and 5 mm Hg to remove low boiling impurities. GC / MS confirmed the structure and purity (about 87%) of 51.9 g (yield 85%) of the title compound.
Step b; Preparation of iodomethylbis (4-methoxyphenyl) methylsilane: 51.9 g (0.169 mol) of chloromethylbis (4-methoxyphenyl) methylsilane in 400 ml of 2-butanone and 25.5 g (0.17 mol) of sodium iodide The solution of was refluxed overnight. This solution was filtered and evaporated. The resulting liquid was redissolved in 500 ml of ethyl acetate, washed with 3 × 250 ml of water and 3 × 250 ml of saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate, filtered and evaporated. The title compound produced in pale orange was pure (about 87%) to a degree sufficient to use as is.
Step c; Preparation of 2,6-di-t-butyl-4-[(dimethyl-di-p-methoxyphenylsilyl) methyloxy] phenol: iodomethylbis (4-methoxyphenyl) methyl in 500 ml of anhydrous acetonitrile A solution of 53.7 g (0.135 mol) of silane and 30.0 g (0.135 mol) of 2,6-di-t-butylbenzhydroquinone was completely degassed with nitrogen. To this solution was added 20.0 g (0.145 mol) of potassium carbonate and the mixture was refluxed under nitrogen for 3 days. Since only traces of product appeared in the subsequent GC, 2.0 g of lithium bromide was added and refluxing continued overnight. About 10% of the product appeared in GC, and lithium bromide was added twice more once a day. 2.0 g of cesium carbonate was also added at this interval. After a total of 15 days of reflux, the reaction mixture was still considered to be about 30% product. The reaction mixture was cooled down, filtered and evaporated. The resulting oil was redissolved in 500 ml of ethyl acetate, washed with 3 x 250 ml of water and 3 x 250 ml of saturated sodium chloride, dried over anhydrous magnesium sulfate and evaporated. The resulting yellow oil crystallized when left. After treating this solid with methanol, it recrystallized from methanol and obtained the title compound as 15.8g (yield 24%) of white solids of melting | fusing point 131-133 degreeC.
Elemental analysis for C ₃₀ H ₄₀ O ₄ Si: Calcd: C, 73.13; H, 8. 18; Found: C, 73.14; H, 8.20.
NMR (CDCl ₃ ): 7.54 (d, 4H, J = 8.5), 6.92 (d, 4H, J = 8.5), 6.82 (s, 2H), 4.73 (s, 1H), 3.96 (s 2H), 3.81 ( s, 6H), 1.42 (s, 18H), 0.64 (s, 3H).
<Example 24>
2-t-butyl-4-[(dimethylbenzylsilyl) methyloxy] phenol (MDL 108,804)
Step a; Preparation of Dimethylbenzylchloromethylsilane: A solution of 68.4 g (0.4 mol) of benzylbromide in 400 ml of THF was added to a suspension of 9.7 g (0.4 mol) of "activated" magnesium in 100 ml of dry THF containing iodine crystals. A solution of 52.7 mL (0.4 mol) of dimethylchloromethylsilane in 200 mL of dry THF was added by the method of step a of Example 2 to obtain the title compound (yield 67%) having a boiling point of 60 to 80 ° C. at 5 mmHg.
Elemental Analysis for C ₉ H ₁₅ ClOSi: Calcd: C, 60.43; H, 7.61; Found: C, 60.29; H, 7.77.
Step b; Preparation of 2-t-butyl-4-[(dimethylbenzylsilyl) methyloxy] phenol (MDL 108,804): 55.2 g (0.3 mol) of dimethylbenzylchloromethylsilane and 600 g (0.35 mol) of sodium iodide in 600 ml of acetonitrile The mixture of was heated at reflux for 24 hours, 20 ml of solvent was distilled off to remove any water and the mixture was cooled to ambient temperature. To the cooled mixture was added 49.8 g (0.3 mol) of 2-t-butylbenzhydroquinone and 50 g (0.15 mol) of cesium carbonate. This mixture was heated for 3 days at 85-90 ° C. under inert atmosphere, cooled and poured into a mixture of water / ethyl acetate (1 L each). The organic layer was isolated, dried and evaporated. The residue was heated on 90 ° C. for 3 hours on a Kugelroer apparatus. The residue was chromatographed (twice with 9/1 hexanes / ethyl acetate, then once with 1/1). The residue was 10.5 g (10%) liquid.
Elemental analysis for C ₂₀ H ₂₈ O ₂ Si: Calcd: C, 73.12; H, 8.59; Found: C, 72.60; H, 8.89.
The following compounds can be prepared by methods analogous to those described in Examples 1-24 above.
2,5-di-t-butyl-4-[(triethylsilyl) methylthio] phenol,
2,5-di-t-butyl-4-[(diethylphenylsilyl) methylthio] phenol,
2,5-di-t-butyl-4-[(tripropylsilyl) methylthio] phenol,
2,5-di-t-butyl-4-[(dipropylphenylsilyl) methylthio] phenol,
2,5-di-t-butyl-4-[(triisopropylsilyl) methylthio] phenol,
2,5-di-t-butyl-4-[(diisopropylphenylsilyl) methylthio] phenol,
2,5-di-t-butyl-4-[(tributylsilyl) methylthio] phenol,
2,5-di-t-butyl-4-[(dibutylphenylsilyl) methylthio] phenol,
2,5-di-t-butyl-4-[(triisobutylsilyl) methylthio] phenol,
2,5-di-t-butyl-4-[(diisobutylphenylsilyl) methylthio] phenol,
2,5-di-t-butyl-4-[(tri-t-butylsilyl) methylthio] phenol,
2,5-di-t-butyl-4-[(di-t-butylphenylsilyl) methylthio] phenol,
2,5-di-methyl-4-[(trimethylsilyl) methylthio] phenol,
2,5-di-methyl-4-[(dimethylphenylsilyl) methylthio] phenol,
2,5-di-methyl-4-[(dibutylphenylsilyl) methylthio] phenol,
2,5-di-methyl-4-[(tri-t-butylsilyl) methylthio] phenol,
2,5-di-methyl-4-[(di-t-butylphenylsilyl) methylthio] phenol,
2,5-di-ethyl-4-[(trimethylsilyl) methylthio] phenol,
2,5-di-ethyl-4-[(dimethylphenylsilyl) methylthio] phenol,
2,5-di-ethyl-4-[(tri-t-butylsilyl) methylthio] phenol,
2,5-di-ethyl-4-[(di-t-butylphenylsilyl) methylthio] phenol,
2,5-di-propyl-4-[(trimethylsilyl) methylthio] phenol,
2,5-di-propyl-4-[(dimethylphenylsilyl) methylthio] phenol,
2,5-di-isopropyl-4-[(trimethylsilyl) methylthio] phenol,
2,5-di-isopropyl-4-[(dimethylphenylsilyl) methylthio] phenol,
2,5-di-butyl-4-[(trimethylsilyl) methylthio] phenol,
2,5-di-butyl-4-[(dimethylphenylsilyl) methylthio] phenol,
2,5-dimethyl-4-[(trimethylsilyl) methyloxy] phenol,
2,5-dimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol,
2,5-dibutyl-4-[(triethylsilyl) methyloxy] phenol,
2,5-dibutyl-4-[(diethylphenylsilyl) methyloxy] phenol,
2-t-butyl-4-[(triethylsilyl) methylthio] phenol,
2-t-butyl-4-[(diethylphenylsilyl) methylthio] phenol,
2-t-butyl-4-[(tripropylsilyl) methylthio] phenol,
2-t-butyl-4-[(dipropylphenylsilyl) methylthio] phenol,
2-t-butyl-4-[(triisopropylsilyl) methylthio] phenol,
2-t-butyl-4-[(diisopropylphenylsilyl) methylthio] phenol,
2-t-butyl-4-[(tributylsilyl) methylthio] phenol,
2-t-butyl-4-[(dibutylphenylsilyl) methylthio] phenol,
2-t-butyl-4-[(triisobutylsilyl) methylthio] phenol,
2-t-butyl-4-[(diisobutylphenylsilyl) methylthio] phenol,
2-t-butyl-4-[(tri-t-butylsilyl) methylthio] phenol,
2-t-butyl-4-[(di-t-butylphenylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(triethylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(diethylphenylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(tripropylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(dipropylphenylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(triisopropylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(diisopropylphenylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(tributylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(dibutylphenylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(triisobutylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(diisobutylphenylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(tri-t-butylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(di-t-butylphenylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(4-aminophenyldimethylsilyl) methyloxy] phenol,
2,3,5-trimethyl-4-[(triethylsilyl) methylthio] phenol,
2,3,5-trimethyl-4-[(diethylphenylsilyl) methylthio] phenol,
2,3,5-trimethyl-4-[(tripropylsilyl) methylthio] phenol,
2,3,5-trimethyl-4-[(dipropylphenylsilyl) methylthio] phenol,
2,3,5-trimethyl-4-[(triisopropylsilyl) methylthio] phenol,
2,6-di-t-butyl-4-[(4-N-methylaminophenyldimethylsilyl) methyloxy] phenol,
2,6-di-t-butyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) methyloxy] phenol,
2,6-di-t-butyl-4-[(4-aminophenyldimethylsilyl) methylthio] phenol,
2,6-di-t-butyl-4-[(4-N-methylaminophenyldimethylsilyl) methylthio] phenol,
2,6-di-t-butyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) methylthio] phenol,
2,5-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol,
2,5-di-t-butyl-4-[(4-aminophenyldimethylsilyl) methyloxy] phenol,
2,5-di-t-butyl-4-[(4-N-methylaminophenyldimethylsilyl) methyloxy] phenol,
2,5-di-t-butyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) methyloxy] phenol,
2,5-di-t-butyl-4-[(4-aminophenyldimethylsilyl) methylthio] phenol,
2,5-di-t-butyl-4-[(4-N-methylaminophenyldimethylsilyl) methylthio] phenol,
2,5-di-t-butyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) methylthio] phenol,
2-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol,
2-t-butyl-4-[(4-aminophenyldimethylsilyl) methyloxy] phenol,
2-t-butyl-4-[(4-N-methylaminophenyldimethylsilyl) methyloxy] phenol,
2-t-butyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) methyloxy] phenol,
2-t-butyl-4-[(4-aminophenyldimethylsilyl) methylthio] phenol,
2-t-butyl-4-[(4-N-methylaminophenyldimethylsilyl) methylthio] phenol,
2-t-butyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol,
2,3,6-trimethyl-4-[(4-aminophenyldimethylsilyl) methyloxy] phenol,
2,3,6-trimethyl-4-[(4-N-methylaminophenyldimethylsilyl) methyloxy] phenol,
2,3,6-trimethyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) methyloxy] phenol,
2,3,6-trimethyl-4-[(4-aminophenyldimethylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(4-N-methylaminophenyldimethylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) methylthio] phenol,
2,3,5-trimethyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol,
2,3,5-trimethyl-4-[(4-aminophenyldimethylsilyl) methyloxy] phenol,
2,3,5-trimethyl-4-[(4-N-methylaminophenyldimethylsilyl) methyloxy] phenol,
2,3,5-trimethyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) methyloxy] phenol,
2,3,5-trimethyl-4-[(4-aminophenyldimethylsilyl) methylthio] phenol,
2,3,5-trimethyl-4-[(4-N-methylaminophenyldimethylsilyl) methylthio] phenol,
2,3,5-trimethyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) methylthio] phenol,
2,3,5-trimethyl-4-[(diphenylmethylsilyl) methyloxy] phenol,
2,6-di-t-butyl-4-[(dimethyl-4-trifluoromethylphenylsilyl) methyloxy] phenol,
2,3,6-trimethyl-4-[(dimethyl-4-trifluoromethylphenylsilyl) methyloxy] phenol,
2,6-di-t-butyl-4-[(dimethyl-3-trifluoromethylphenylsilyl) methyloxy] phenol,
2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol propionic acid ester,
2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol butyric acid ester,
2,6-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol acetic acid ester,
2,6-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol succinic acid ester,
2,6-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol butyric acid ester,
2,6-di-t-butyl-4-[(4-N-methylaminophenyldimethylsilyl) methyloxy] phenol acetic acid ester,
2,6-di-t-butyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) methyloxy] phenol succinic acid ester,
2,6-di-t-butyl-4-[(4-aminophenyldimethylsilyl) methylthio] phenol acetic acid ester,
2,6-di-t-butyl-4-[(4-N-methylaminophenyldimethylsilyl) methylthio] phenol succinic acid ester,
2,6-di-t-butyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) methylthio] phenol propionic acid ester,
2,5-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol acetic acid ester,
2,5-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol succinic acid ester,
2,5-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol propionic acid ester,
2,3,6-trimethyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol butyric acid ester,
2,3,6-trimethyl-4-[(4-N-methylaminophenyldimethylsilyl) methyloxy] phenol acetic acid ester,
2,3,6-trimethyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) methyloxy] phenol succinic acid ester,
2,3,5-trimethyl-4-[(4-aminophenyldimethylsilyl) methylthio] phenol acetic acid ester,
2,3,5-trimethyl-4-[(4-N-methylaminophenyldimethylsilyl) methylthio] phenol succinic acid ester,
2,3,5-trimethyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) methylthio] phenol propionic acid ester,
2,5-di-t-butyl-4-[(triethylsilyl) methylthio] phenol acetic acid ester,
2,5-di-t-butyl-4-[(diethylphenylsilyl) methylthio] phenol succinic acid ester,
2,5-di-t-butyl-4-[(tripropylsilyl) methylthio] phenol acetic acid ester,
2,5-di-t-butyl-4-[(dipropylphenylsilyl) methylthio] phenol acetic acid ester,
2,5-di-t-butyl-4-[(triisopropylsilyl) methylthio] phenol propionic acid ester,
2,5-di-t-butyl-4-[(diisopropylphenylsilyl) methylthio] phenol butyric acid ester,
2,5-di-t-butyl-4-[(tributylsilyl) methylthio] phenol succinic acid ester,
2,5-di-t-butyl-4-[(dibutylphenylsilyl) methylthio] phenol acetic acid ester,
2,5-di-t-butyl-4-[(triisobutylsilyl) methylthio] phenol acetic acid ester,
2,5-di-t-butyl-4-[(diisobutylphenylsilyl) methylthio] phenol succinic acid ester,
2,5-di-t-butyl-4-[(tri-t-butylsilyl) methylthio] phenol succinic acid ester,
2,5-di-t-butyl-4-[(di-t-butylphenylsilyl) methylthio] phenol acetic acid ester,
2,3,6-trimethyl-4-[(diphenylmethylsilyl) methyloxy] phenol acetic acid ester,
2,3,5-trimethyl-4-[(diphenylmethylsilyl) methyloxy] phenol acetic acid ester,
2,6-di-t-butyl-4-[(dimethyl-4-trifluoromethylphenylsilyl) methyloxy] phenol acetic acid ester,
2,3,6-trimethyl-4-[(dimethyl-4-trifluoromethylphenylsilyl) methyloxy] phenol acetic acid ester, and
2,6-di-t-butyl-4-[(dimethyl-3-trifluoromethylphenylsilyl) methyloxy] phenol acetic acid ester.
General synthetic schemes for preparing compounds of formula (1) wherein Z is methylene are described in Scheme B, with all substituents in the formula being as defined above unless otherwise noted.
In general, the phenol of structure 1c can be prepared according to Scheme B in two steps. In step a, haloalkensilanes of suitable structure 3 are reacted with magnesium metal in a suitable protic solvent such as ethyl ether to produce a magnesium halide salt. The magnesium halide salt (Grignard reagent) is then reacted with the appropriate alkyl-4-hydroxy-benzaldehyde (or suitably protected derivative) of structure 4 to obtain an alcohol of structure 5. In step b, the alcohol of structure 5 can be reduced to the desired phenol of structure 1b by various reduction reaction techniques and methods well known and recognized in the art. For example, the alcohol of structure 5 can be reduced by a Birch reduction reaction by reacting with sodium in liquid ammonia.
The phenol esters of structure 1d can be prepared by acylating the phenols of structure 1c according to standard acylation reaction techniques as described above in Scheme A.
Starting materials used in the general synthetic methods outlined in Scheme B can be readily purchased or can be readily prepared according to standard techniques and methods. If it is necessary to prevent undesired side reactions, the 1-phenol functionality of the alkyl-4-hydroxy-benzaldehyde of structure 4 in Scheme B may be blocked prior to Grignard reaction with a standard phenol blocker as described above in Scheme A. have.
The following examples show representative synthesis examples described in Scheme B. Such embodiments are to be understood as illustrative only and are not intended to limit the scope of the invention in any way.
<Example 25>
2,3,6-dimethyl-4- [2- (trimethylsilyl) ethyl] phenol
Step a; Magnesium turning 240 mg (10 mmol) and anhydrous ethyl ether were mixed under inert atmosphere. A solution of 1.9 g (10 mmol) of chloromethyltrimethylsilane in anhydrous ethyl ether was added. Stir until magnesium metal is dissolved. A solution of 1.7 g (10 mmol) of 2,3,5-trimethyl-4-hydroxybenzaldehyde 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 give 4-hydroxy-2,3,5-trimethyl-α-[(trimethylsilyl) methyl] benzenemethanol and purified by silica gel chromatography.
Step b; 520 mg (22.6 mmol) of sodium metal and 13 ml of liquid ammonia were mixed. To this solution was added dropwise a solution of 2.37 g (10 mmol) of 4-hydroxy-2,3,5-trimethyl-α-[(trimethylsilyl) methyl] benzenemethanol in 0.5 g of ethyl alcohol and 5 ml of ethyl ether. After the blue disappeared, 13 ml of water were carefully added, extracted with ethyl ether, dried (MgSO ₄ ) and the solvent was evaporated. The residue was purified by silica gel chromatography to give the title compound.
Alternatively, the compound of formula (1) wherein Z is methylene can be prepared according to the method described in Scheme C, and all substituents in the scheme are as already described unless otherwise stated.
In general, the phenol of structure 1b can be prepared by first reacting a suitable haloalkylenesilane of structure 3 with magnesium metal in a suitable protic solvent such as ethyl ether to produce a magnesium halide salt. The magnesium halide salt (Grignard reagent) is then reacted with the appropriate alkyl-4-hydroxy-benzylhalide (or suitably protected derivative) of structure 6 to obtain the desired phenol of structure 1c.
The phenol esters of structure 1d can be prepared by acylating the phenols of structure 1c according to standard acylation reaction techniques as described above in Scheme A.
Starting materials used in the general synthetic methods outlined in Scheme C can be readily purchased or can be readily prepared according to standard techniques and methods. For example, 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 phenolic starting material by standard hydrolysis methods.
If it is necessary to prevent undesired side reactions, the 1-phenol functionality of the alkyl-4-hydroxy-benzylhalide of structure 6 in Scheme C should be blocked prior to Grignard reaction with a standard phenol blocker as described above in Scheme A. Can be.
The following examples show representative synthesis examples described in Scheme C. These examples are to be understood as illustrative only and are not intended to limit the scope of the invention in any way.
Example 26
2,6-diethyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) ethyl] phenol
Magnesium turning 240 mg (10 mmol) and anhydrous ethyl ether were mixed under inert atmosphere. A solution of 3.19 g (10 mmol) of 4-N, N-dimethylaminophenyl (dimethyl) iodomethylsilane (Example 2) in anhydrous ethyl ether was added. Stir until magnesium metal is dissolved. A solution of 2.43 g (10 mmol) of 4-bromomethyl-2,6-diethylphenol in anhydrous ethyl ether was added and the mixture was refluxed until the reaction was complete. It 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.
Example 27
2,6-diethyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) ethyl] phenol acetic acid ester
7.1 g (20 mmol) of 2,6-diethyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) ethyl] phenol (Example 26) in 150 ml of ether and 2.53 g (25 mmol) of triethylamine ) Was stirred at room temperature. 1.96 g (25 mmol) of acetyl chloride were added and the mixture was stirred overnight. Water and ether were added and the layers separated. The organic layer was evaporated to give an oil which was distilled in kugelloar. Chromatography (chloroform) on silica gel gave the title compound.
<Example 28>
2,6-di-t-butyl-4-[(diphenylmethylsilyl) ethyl] phenol
A solution of 1.85 g (10 mmol) of diphenyl (methyl) chloromethylsilane (Example 1) in anhydrous ethyl ether was mixed in 240 mg (10 mmol) of magnesium turning and anhydrous ethyl ether in a similar manner to that described in Example 26. And a solution of 2.9 g (10 mmol) of 4-bromomethyl-2,6-di-t-butylphenol (Maybridge # MB 00185) in anhydrous ethyl ether to give the title compound.
The following compounds can be prepared by methods analogous to those described in Examples 25-28 above.
2,5-dipropyl-4- [2- (trimethylsilyl) ethyl] phenol,
2,5-dipropyl-4- [2- (dimethylphenylsilyl) ethyl] phenol,
2,5-diisopropyl-4- [2- (trimethylsilyl) ethyl] phenol,
2,5-diisopropyl-4- [2- (dimethylphenylsilyl) ethyl] phenol,
2,5-diisobutyl-4- [2- (trimethylsilyl) ethyl] phenol,
2,5-diisobutyl-4- [2- (dimethylphenylsilyl) ethyl] phenol,
2,5-dibutyl-4- [2- (trimethylsilyl) ethyl] phenol,
2,5-dibutyl-4- [2- (dimethylphenylsilyl) ethyl] phenol,
2,5-di-t-butyl-4- [2- (trimethylsilyl) ethyl] phenol,
2,5-di-t-butyl-4- [2- (dimethylphenylsilyl) ethyl] phenol,
2,5-di-t-butyl-4- [2- (tri-t-butylsilyl) ethyl] phenol,
2,5-di-t-butyl-4- [2- (di-t-butylphenylsilyl) ethyl] phenol,
2,5-dimethyl-4- [2- (trimethylsilyl) ethyl] phenol,
2,5-dimethyl-4- [2- (dimethylphenylsilyl) ethyl] phenol,
2-t-butyl-4- [2- (trimethylsilyl) ethyl] phenol,
2-t-butyl-4- [2- (dimethylphenylsilyl) ethyl] phenol,
2-t-butyl-4- [2- (trimethylsilyl) ethyl] phenol,
2-t-butyl-4- [2- (dimethylphenylsilyl) ethyl] phenol,
2-t-butyl-4- [2- (trimethylsilyl) ethyl] phenol,
2-t-butyl-4- [2- (dimethylphenylsilyl) ethyl] phenol,
2-t-butyl-4- [2- (trimethylsilyl) ethyl] phenol,
2-t-butyl-4- [2- (dimethylphenylsilyl) ethyl] phenol,
2,3,5-tri-t-butyl-4- [2- (trimethylsilyl) ethyl] phenol,
2,3,5-tri-t-butyl-4- [2- (dimethylphenylsilyl) ethyl] phenol,
2,3,5-tri-t-butyl-4- [2- (tri-t-butylsilyl) ethyl] phenol,
2,3,5-tri-t-butyl-4- [2- (di-t-butylphenylsilyl) ethyl] phenol,
2,3,6-trimethyl-4- [2- (trimethylsilyl) ethyl] phenol,
2,3,6-trimethyl-4- [2- (dimethylphenylsilyl) ethyl] phenol,
2,6-di-t-butyl-4-[(4-N-methylaminophenyldimethylsilyl) ethyl] phenol,
2,6-di-t-butyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) ethyl] phenol,
2,6-di-t-butyl-4-[(4-aminophenyldimethylsilyl) ethyl] phenol,
2,5-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) ethyl] phenol,
2,5-di-t-butyl-4-[(4-aminophenyldimethylsilyl) ethyl] phenol,
2,5-di-t-butyl-4-[(4-N-methylaminophenyldimethylsilyl) ethyl] phenol,
2,5-di-t-butyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) ethyl] phenol,
2,6-di-t-butyl-4-[(dimethylphenylsilyl) ethyl] phenol propionic acid ester,
2,6-di-t-butyl-4-[(dimethylphenylsilyl) ethyl] phenol butyric acid ester,
2,6-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) ethyl] phenol acetic acid ester,
2,6-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) ethyl] phenol succinic acid ester,
2,6-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) ethyl] phenol butyric acid ester,
2,6-di-t-butyl-4-[(4-N-methylaminophenyldimethylsilyl) ethyl] phenol acetic acid ester,
2,6-di-t-butyl-4-[(4-N-methyl-N-ethylaminophenyldimethylsilyl) ethyl] phenol succinic acid ester,
2,6-di-t-butyl-4-[(4-aminophenyldimethylsilyl) ethyl] phenol acetic acid ester,
2,5-di-t-butyl-4-[(triethylsilyl) ethyl] phenol acetic acid ester,
2,5-di-t-butyl-4-[(diethylphenylsilyl) ethyl] phenol succinic acid ester,
2,3,6-trimethyl-4-[(diphenylmethylsilyl) methyloxy] phenol acetic acid ester,
2,3,5-trimethyl-4-[(diphenylmethylsilyl) methyloxy] phenol acetic acid ester,
2,6-di-t-butyl-4-[(dimethyl-4-trifluoromethylphenylsilyl) methyloxy] phenol acetic acid ester,
2,3,6-trimethyl-4-[(dimethyl-4-trifluoromethylphenylsilyl) methyloxy] phenol acetic acid ester, and
2,6-di-t-butyl-4-[(dimethyl-3-trifluoromethylphenylsilyl) methyloxy] phenol succinic acid ester.
It is contemplated that the compounds of formula (1) may exist in various stereoisomeric forms. All stereoisomeric forms consistent with the above formulas (interpreted in accordance with standard practice of expressing stereoisomeric structures) are intended to be included within the scope of the present invention.
Preferred compounds of formula (1) are those in which R is hydrogen, acetyl or succinyl; R ₁ is methyl or tertiary butyl; R ₂ and R ₃ are each independently hydrogen, methyl or tertiary butyl; R ₄ is hydrogen or methyl; R ₆ is methyl; A is methylene; R ₅ and R ₇ are each independently methyl or — (CH ₂ ) _n — (Ar) where n is 0 or 1 and Ar is hydroxy, methoxy, ethoxy, halogen, trifluoromethyl, C _{Is 1} -C ₆ alkyl or -NR ₈ R ₉ , wherein R ₈ and R ₉ are each independently phenyl unsubstituted or substituted with one to three substituents selected from the group consisting of hydrogen or methyl . More preferable are the following compounds.
2,6-di-t-butyl-4-[(diphenylmethylsilyl) methyloxy] phenol,
2,6-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol,
2,6-di-t-butyl-4-[(dimethyl-4-trifluoromethylphenylsilyl) methyloxy] phenol,
2,6-di-t-butyl-4-[(dimethyl-3-trifluoromethylphenylsilyl) methyloxy] phenol,
2-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol,
2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol succinic acid ester,
2-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol succinic acid ester,
2,6-di-t-butyl-4-[(dimethylphenylsilyl) methylthio] phenol succinic acid ester,
2,6-di-t-butyl-4-[(trimethylphenylsilyl) methylthio] phenol succinic acid ester,
2-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol acetic acid ester,
2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol acetic acid ester,
2,3,6-trimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol acetic acid ester,
2,5-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol,
2,5-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol acetic acid ester,
2-t-butyl-4-[(dimethylphenylsilyl) methylthio] phenol,
2,3,6-trimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol,
2,3,5-trimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol,
2-t-butyl-4-[(dimethyl-p-methoxyphenylsilyl) methyloxy] phenol,
2,5-di-t-butyl-4-[(diphenylmethylsilyl) methyloxy] phenol,
2,6-di-t-butyl-4-[(diphenylmethylsilyl) methyloxy] phenol,
2,6-di-t-butyl-4-[(methyl-di-p-methoxyphenylsilyl) methyloxy] phenol,
2,6-di-t-butyl-4-[(dimethyl-p-methoxybenzylsilyl) methyloxy] phenol, and
2-t-butyl-4-[(dimethylbenzylsilyl) methyloxy] phenol.
The term “patient” as used herein requires the treatment of chronic inflammatory disease, atherosclerosis, hypercholesterolemia, or cytokine-induced expression of vascular cell adsorption molecule-1 and / or intercellular adsorption molecule-1. Refers to a warm-blooded animal or mammal in need of inhibition. Guinea pigs, dogs, cats, rats, mice, hamsters, rabbits and primate animals, including humans, are considered examples of patients within the scope of this term.
Atherosclerosis is a disease state characterized by the development and increase of atherosclerosis lesions or plaques. Identifying patients in need of treatment of atherosclerosis is by the ability and knowledge of one of ordinary skill in the art. For example, each patient suffering from clinically significant atherosclerosis or at risk of developing clinically significant atherosclerosis is a patient in need of treatment of atherosclerosis. One of ordinary skill in the art can readily determine patients in need of treatment of atherosclerosis using clinical trials, physical examinations and medical and / or family histories.
An atherosclerosis inhibiting effective amount of the compound of formula (1) is an effective amount that inhibits the atherosclerosis in a patient in need of inhibiting the onset or progression of atherosclerosis. As such, successful treatment of atherosclerosis patients is understood to include effectively delaying, stopping, arresting or stopping atherosclerosis lesions or plaques and necessarily atherosclerosis It does not indicate the removal of the whole. In addition, it is understood and recognized by one of ordinary skill in the art that successful treatment of atherosclerosis may include preventive measures that interfere with atherosclerosis lesions or plaque formation.
Peroxidation of LDL lipids, such as the unsaturated fatty acid portion of LDL cholesteryl esters, is known to promote the deposition of cholesterol in macrophages that deposit on the vessel walls and transform into foam cells. Identifying patients in need of inhibition of LDL lipid peroxidation is by the skill and knowledge of one of ordinary skill in the art. For example, each patient in need of treatment of atherosclerosis as defined above is also a patient in need of inhibition of LDL lipid peroxidation. An antioxidant effective amount of the compound of formula (1) is an amount effective to inhibit LDL lipid peroxidation in the blood of a patient.
Hypercholesterolemia is a disease condition characterized by elevated levels of serum cholesterol or LDL cholesterol in clinically significant amounts, beyond what is considered normal by one of ordinary skill in the art. Identifying patients in need of treatment of hypercholesterolemia is by the skill and knowledge of one of ordinary skill in the art. For example, an individual with serum cholesterol levels or LDL cholesterol levels determined in clinical trials, substantially and clinically increased than would be considered normal by one of ordinary skill in the art, may be treated for hypercholesterolemia. It is a patient in need. In addition, for example, an individual at risk of developing hypercholesterolemia may also be a patient in need of treatment of hypercholesterolemia. One of ordinary skill in the art will use patients who are at risk of developing hypercholesterolemia and patients with hypercholesterolemia who require treatment of hypercholesterolemia using clinical trials, physical examinations, medicine and / or family history. It's easy to decide.
The term "chronic inflammatory disease" refers to a disease or condition characterized by persistent inflammation in the absence of irritating irritants or microbial pathogens. Inflammatory diseases treated with compounds of formula (1) include, in particular, asthma, chronic inflammation, rheumatoid arthritis, autoimmune diabetes, graft rejection and tumor angiogenesis. A “therapeutically effective amount” of a compound of formula (1) is an effective amount according to single or multiple administrations to a patient that alleviates the symptoms associated with a chronic inflammatory disease. The "effective amount of vascular cell adsorption molecule-1 and / or intercellular cell adsorption molecule-1 inhibition" of the compound of formula (1) is a symptom associated with the vascular cell adsorption molecule-1 and / or intercellular adsorption molecule-1 mediated state. Effective amount according to single or multiple administrations to a patient.
As used herein, "mitigating symptoms" of a chronic inflammatory disease or vascular cell adsorption molecule-1 mediated condition means reducing the severity expected without treatment and does not necessarily indicate eliminating or treating the entire disease. Does not.
To determine a therapeutically effective amount or dosage of Formula (1), an antioxidant effective amount or dosage, a plasma cholesterol lowering dosage or dosage, an atherosclerosis inhibiting effective amount or dosage, or VCAM-1 and / or ICAM-1 inhibitory effective amount There are a number of factors (species of mammals; their size, age and overall health; the specific disease involved; the severity or incidence of the disease; the response of the individual patient; the specific compound administered; Concomitant dosing regimens; concomitant use of drug therapy; and other related situations) are contemplated by the attending physician.
A therapeutically effective amount or dose of Formula (1), an antioxidant inhibitory dose or dose, a plasma cholesterol lowering dose or dose, an atherosclerosis inhibiting effective amount or dosage, or VCAM-1 and / or ICAM-1 inhibitory effective amount is generally From about 1 mg / kg body weight per day (mg / kg / day) to about 5 g / kg body weight per day (g / kg / 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 and / or ICAM-1 expression. The compounds of the present invention exhibit an inhibitory effect through the inhibition of VCAM-1 and / or ICAM-1 upregulation by cytokines, thereby resulting in chronic inflammatory, including asthma, chronic inflammation, rheumatoid arthritis, autoimmune diabetes, and the like. disease; It is thought to prevent or alleviate the symptoms of atherosclerosis and hypercholesterolemia. However, it is to be understood that the present invention is not limited by any particular theory or proposed mechanism to explain its utility in the intended use.
In treating a patient, the compound of formula (1) may be administered in any form or manner which makes the compound effective in vivo, including oral and parenteral routes. 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 art of making formulations can readily select the appropriate dosage form and mode, depending on the disease state to be treated, the stage of the disease and other relevant circumstances (Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Co. (1990). )).
The compound of formula (1) may be administered in the form of a pharmaceutical composition or medicament prepared by combining the compound of formula (1) with a pharmaceutically acceptable carrier or excipient, the proportions and properties of which are selected by the route of administration and standard agent Determined by a chemical practice.
Pharmaceutical compositions or medicaments are prepared by methods well known in the art of pharmacy. The carrier or excipient may be a solid, semisolid or liquid substance which can act as a vehicle or medium of the active ingredient. Suitable carriers or excipients are well known in the art. The pharmaceutical compositions can be adapted for oral or parenteral use and can be administered to the patient in the form of tablets, capsules, suppositories, solutions, suspensions and the like.
The pharmaceutical composition can be administered orally with an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For oral therapeutic 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% of the compound of formula (1) as the active ingredient, but can vary depending on the particular form and can conveniently be from 4% to about 70% of the unit's body weight. The amount of the active ingredient present in the composition is that amount in which a unit dosage form suitable for administration is obtained.
Tablets, pills, capsules, troches, and the like may be combined with one or more adjuvants such as microcrystalline cellulose, gum tragacanth or gelatin; Excipients such as starch or lactose; Disintegrants such as alginic acid, Primogel, corn starch and the like; 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 salicylate or orange spice. When the dosage unit form is a capsule, it may contain, in addition to substances of this type, liquid carriers such as polyethylene glycol or fatty oils. Other dosage unit forms may contain various other materials, such as coatings, that modify the material form of the dosage unit. Thus, tablets or pills can be coated with sugars, shellac or other enteric coatings. Syrups may contain sucrose and certain preservatives, dyes, colorants and flavoring agents as sweeteners in addition to the active ingredient. The materials used to prepare these various compositions must be pharmaceutically pure and nontoxic when used.
For parenteral administration, the compound of formula (1) may be mixed in solution or suspension. Such formulations should contain at least 0.1% of the compounds of the present invention, but can vary from 0.1 to about 50% of body weight. The amount of active ingredient present in such compositions is that amount in which appropriate administration is made.
The solution or suspension may be formulated with one or more auxiliaries, such as water for injection, physiological saline, nonvolatile oils, polyethylene glycol, glycerin, propylene glycol or other synthetic solvents, depending on the solubility and other properties of the compound of formula (1). Sterile diluents; Bactericides 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 acetates, citrate or phosphates; And toxicity modulators such as sodium chloride or dextrose.
<Example 29>
Cell Surface ELISA for VCAM-1 and / or ICAM-1
Proliferating human umbilical vein endothelial cells (HUVEC) or human aortic smooth muscle cells (HASMC) from Clonetics, San Diego, Calif., Play at 20,000 cells per cm 2 in 100 μl of medium per well on 96-well plates. Ting. Cultures were maintained for 2 days in growth medium (EGM or SMGM2, Clontics, San Diego, Calif.) Before adding cytokines or drugs. Compound-containing or non-cytokine was 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, Gaithersburg, MD) was added to the embryos at 100-200 pg / ml (addition of 100 μl of cytokine containing compound aseptically diluted on each 96-well plate By transfer into cells containing cells, culture medium was not exchanged before adding effector). The culture medium was removed and the monolayers were washed twice with Hanks buffered saline (HBSS) at room temperature. Anti-human VCAM-1 or Immuntech, Inc., Westbrook, ME of primary antibody (Upstate Biotechnology, Inc., Lake Placid, NY) Human ICAM-1) was added to each well and incubated at 37 ° C. for 1 hour at 37 ° C., with new calf serum (Gibco-BRL, Gaithersburg, MD) containing 5% HBSS. The wells were washed twice with HBSS and then 1/1000 dilution of goat anti-mouse IgG (BioRad, Hercules, CA) bound to horseradish peroxidase in HBSS containing 5% newborn calf serum. 100 μl was added to each well and incubated at 37 ° C. for 1 hour. Wells were washed three times with HBSS, then 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 at 450 nm with a plate reader. IC ₅₀ values were determined from curves of absorbance values obtained from serial dilutions of compounds (dissolved in dimethyl sulfoxide).
IC ₅₀ values are defined as drug concentrations that inhibit cytokine-induced adsorption molecule expression by 50%. The maximum value of adsorption molecule expression in the cytokine-induced culture was determined by subtracting the basal level of adsorption molecule expression in the culture (without cytokine) to determine the level of induction. VCAM-1 was usually induced at about 5-7 fold. ICAM-1 was induced approximately 5-10 fold. Each drug concentration was tested four times in the wells. Single point testing of compounds at 50 μM was assessed as described in IC ₅₀ determinations, except that the data showed inhibition levels without correction of baseline expression (baseline adsorption molecule expression was 10-20 of total induced expression. %).
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.
Inhibition of VCAM-1 in Human Aortic Smooth Muscle Cells (HASMC) Compound Number (MDL Number)HSMC-1 (% inhibition, 50 μM)HSMC-2 (% inhibition, 50 μM)HSMC-3 (% inhibition, 50 μM)VCAM-1 (average) 104,59950.138.049.045.7 104,55654.158.058.056.7 105,975(11.6)20.051.019.8 103,491N.T. *57.049.053.0 103,07630.655.046.043.9 103,14113.747.033.031.2 104,86356.552.056.054.8 103,377N.T.45.046.045.5 105,44311.444.022.025.8 105,3148.738.038.028.2 103,65363.154.052.056.4 * N.T. = not tested
Table 2 summarizes the ability of the various compounds of the invention to selectively inhibit VCAM-1 or to inhibit both VCAM-1 and ICAM-1 using proliferating human umbilical vein endothelial cells (HUVEC). In this experiment, cells were co-cultured with tumor necrosis factor-α for 20-24 hours with the indicated compounds prior to assessing cell surface adsorption molecule expression.
Inhibition of VCAM-1 and / or ICAM-1 in Human Vein Endothelial Cells (HUVEC) Compound Number (MDL Number)VCAM-1 (% inhibition, 50 μM) *ICAM-1 (% inhibition, 50 μM) @ 104,59912.3(9.5) 104,55633.31.5 105,9756.32.5 103,49112.080 103,0767.376 103,1411.379.5 104,86344.353 103,37718.3(5.0) 105,4433.073.0 105,31410.775.5 103,65337.778.5 * Average of 3 trials @ Average of 2 trials, numbers in parentheses indicate negative values
In vivo activity of such compounds can also be assessed in other models of inflammation described above as being associated with increased VCAM-1 levels. One such model for respiratory diseases such as asthma is an ovalbumin sensitized model (Kung, T. T. et al, Int. Arch. Allergy Immunol. 105, 83-90 (1994)). This model of lung inflammation is IgG mediated and is associated with eosinophilia (as in asthmatic human patients). Bronchoalveolar lavage (BAL) fluids obtained from experimental animals can be assessed for a number of variables including soluble adsorption molecule expression and leukocyte accumulation. Adsorption molecule expression can be assessed by immunohistochemistry in the tissues of experimental animals, particularly the lungs. The effect of the claimed compounds, such as MDL 29,353, should be to inhibit the increase in VCAM-1 expression and to inhibit eosinophil accumulation in BAL fluid. Inhibitors can be tested in a rat model of adjuvant arthritis, which has already been shown to respond to anti-ICAM-1 monoclonal antibodies (Iigo, Y. et al., J. Immunol. 147, 4167 -4171 (1991)). In this model, adsorption molecule expression is evaluated in the limbs (joints) of experimental animals. For autoimmune diabetes, the ability of those compounds to delay onset or prevent application metastasis can be tested in NOD mouse models (Heinke, EW et al., Diabetes 42, 1721-1730 (1993); Baron, JL et al., J. Clin. Invest. 93, 1700-1708 (1994)). Furthermore, it is possible to monitor the onset of diabetes in experimental animals as well as to monitor the level of VCAM-1 expression in tissues (eg pancreas). The therapeutic potential of transplant rejection can be assessed by monitoring cardiac tag transplantation survival (Balb / c heart implanted in C3H / He recipients) (Isobe, M. et al., J. Immunol. 153, 5810-5818 (1994) ). In vivo administration of anti-VCAM-1 and anti-VLA-4 monoclonal antibodies induces immune suppression against cardiac tagging implants and soluble antigens in this mouse model. The effect of compounds on tumor metastasis and angiogenesis can be assessed in a number of models. These include B16 (murine) and M24met (human) melanoma models for experimental metastasis (Fidler, IJ, Cancer Res. 35, 218-224 (1975); Meuller, BM et al., Cancer Res. 51, 2193-2198). The activity of the compounds can be assessed not only by the effect on the number of advanced lung metastases, but also by the effect on VCAM-1 expression in the lung as described above in 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 subcutaneous injected basement membrane proteins in mice (Passaniti, A. et al., Lab. Invest. 67, 519-528 (1992)). Angiogenesis is recorded by the number of blood vessels supplemented into matrigel and the hemoglobin content of the gel. Adsorption molecule expression and accumulation of leukocytes can be determined by immunohistochemical methods as in all of the above examples.
<Example 30>
Low Cholesterol and Antioxidant Effects of Compounds of Formula (1) in Cholesterol-fed Female New Zealand White Rabbits
A. Experimental Protocol
Five independent experiments were performed in the following manner. Each study was performed in controls and 1-5 groups treated with MDL compounds (N = 5 per group). Female New Zealand white rabbits (Hazelton, about 2.0 to 2.3 kg) were fed 0.2% cholesterol supplemented rabbit feed (Purina # 5322) with or without 0.4% MDL compound (0.26% for MDL 108,804, 0.6% for MDL 103,491). ). MDL compound was dissolved in 100% ethanol. The MDL compound was not dissolved in 100% ethanol but was dissolved in diethyl ether: ethanol (3: 2, volume ratio). The MDL mixture was sprayed on the food and dried overnight in a chemical vapor hood. Ethanol was injected into the control food. The rabbits were fed 100 g of food per day for 7 days (0.6% MDL 103,491 for 14 days) and the water was not limited. On day 7, approximately 2 ml of blood of the (overnight starved) rabbit was taken from the marginal ear vein; Rabbits treated with 0.6% MDL 103,491 were tested on day 14. Rabbits were euthanized with carbon dioxide overdose. Body and liver weights are reported in grams. Food consumption was recorded in g / day / rabbit. Fresh serum was dispensed and used for clinical chemical analysis in serum, lipoprotein cholesterol measurement, thiobarbituric acid reactive substance (TBARS) evaluation, and determination of compound and metabolite concentrations. The liver (approximately 5 g aliquots) was frozen at −20 ° C. for subsequent determination of compound and metabolite concentrations.
B. Clinical Chemistry Analysis
Blood was allowed to coagulate by placing at room temperature for 30 minutes. Serum was obtained after centrifugation at 3000 rpm and 5 ° C. for 10 minutes using a Beckman GPKR centrifuge equipped with a GH rotor. Fresh serum was analyzed by COBAS MIRA Roche Diagnostics using Roche diagnostic reagents for total cholesterol and triglycerides (CHOL, Kit # 44334; and TG, Kit # 44120, respectively). Cholesterol and triglycerides were calculated as mg / dl.
C. TBARS Assessment
TBARS is a qualitative indicator of lipid oxidation in a sample. In this evaluation, oxidation of serum lipids is initiated with CuSO ₄ , and aldehydes such as malondialdehyde (MDA) are produced. When incubated with thiobarbituric acid, the absorbance of the aldehyde can be detected at 530-540 nm. TBARS values lower than control serum levels indicate the relative ability of compounds to inhibit oxidation. TBARS was determined as follows: 50 μl of serum was mixed with 50 μl of 0.9% physiological saline and 400 μl of 5 mM CuSO ₄ solution and incubated at 37 ° C. for 5 hours. 1.0 ml of 20% trichloroacetic acid was added to stop the reaction. Then, 1.0 ml of 0.67% thiobarbituric acid in 0.05 N sodium hydroxide was added, mixed, and the samples were incubated at 90 ° C. for 30 minutes. Samples were briefly centrifuged to pellet undissolved material and the supernatant transferred to a 96-well microtiter plate. Absorbance was measured at 540 nm using a Biotek model EL311 microplate reader. The resulting MDA (nmol) was calculated from a standard curve of 0-10 nmol of MDA prepared from malonaldehyde bis (dimethylacetal). Serum samples from treated rabbits were compared to serum samples from control rabbits not fed the MDL compound.
D. HPLC quantitation of compound and metabolite concentrations in serum and liver
Serum and liver concentrations of the parent compound, metabolite, bisphenol and diphenoquinone were determined by reverse phase HPLC using a Waters 990 Powerline system. 1 g of liver was homogenized with 5.0 ml of PBS at pH 7.4 for 20-30 seconds at control point 5 using a Polytron tissue homogenizer. Serum or liver homogenate was extracted as follows: 100 μl of serum or homogenate was added to 2.0 ml of diethyl ether: ethanol (3: 1) while vortexing the tube. The sample tube was capped and centrifuged for 10 minutes at 3500 rpm and 5 ° C. with a Beckman GPKR centrifuge equipped with a GH 3.7 rotor. The supernatant was transferred to a clean tube and dried under nitrogen. Samples were reconstituted with 200 μl of acetonitrile: hexanes: 0.1 M ammonium acetate (90: 6.5: 3.5, volume ratio). 100 μl was then injected onto a Waters Deltapak C18-300 mm 3 column and eluted at a flow rate of 1.5 ml / min with 83% acetonitrile: 17% mobile phase. Absorbance was recorded at wavelengths of 240, 254 and 420 nm. Compound concentrations were calculated from known stocks of certain parent compounds after correction for recovery; The recovery range from the spike partial sample ranged from 40 to 100%. The lowest detection limit for this class of compounds was about 0.5 μg / ml. Concentrations were calculated as μg / ml serum and μg / liver g.
E. HPLC Separation and Quantitative Analysis of Lipoprotein Small Fraction Cholesterol Levels
Lipoprotein fractions (ultra low density lipoprotein, VLDL; low density lipoprotein, LDL; and high density lipoprotein, HDL) were separated on a Sepharose 6HR column (1 × 30 cm, Pharmacia) connected to a Waters Powerline HPLC system. 50 μl of serum was injected onto the column and eluted at a flow rate of 0.5 ml / min with phosphate buffered saline at pH 7.4. Cholesterol reagent (Roche Diagnostics, Kit # 44334, diluted with 20 ml of water, then diluted with 20 ml of 0.9% saline solution) was added to the next column eluent at a rate of 0.2 ml / min and knitted PFTE Kratos reaction coil. (Applied Biosystems) incubated for 5 minutes at 37 ℃. Absorbance was measured at 500 nm. Lipoprotein subfractions were quantified as follows.
(Total Serum Cholesterol) × (% Area Under Curve For Each Subfraction)
Tables 3 and 4 below summarize the data from the individual experiments of this test method.
Hypocholesterolemic and Antioxidant Effects of Compounds of Formula (1) in Cholesterol-Fed Female New Zealand White Rabbits as a Percent to Control MDL numberDiet%foodweightlw / bwCHOLLDLHDLTRIGTBARS 103,4910.4100%100%89%85%81%103%134%32% 103,4910.680%95%96%139%ND *ND216%18% 104,5560.4100%97%96%47%53%116%81%70% 104,5990.498%99%86%76%71%105%64%79% 104,9620.469%97%71%118%105%163%159%19% 105,4430.4100%101%90%98%97%115%127%42% 105,9750.4100%101%108%69%77%76%143%68% 106,8340.499%99%94%67%89%61%98%70% 107,9170.4100%99%106%151%165%86%129%68% 108,2080.4100%97%103%109%117%97%107%89% 108,8040.26100%101%96%91%82%109%143%49% * ND = not measured N = 5 rabbits / group; Starved overnight Rabbits were fed for 7 days (exceptionally, MDL 103,491 was fed for 0.6 days at 0.6%) Diet% = (MDL compound weight / feed weight) × (100) Normalize the data in Table 3 as follows: % Control = (treated group average / control average) x (100) feeding = amount fed per rabbit per day (g) body weight = weight gLW / BW = (liver weight g / body weight g) CHOL = total cholesterol (mg / VIII) LDL = low density lipoprotein cholesterol (mg / dl) HDL = high density lipoprotein cholesterol (mg / dl) TRIG = triglycerides (mg / dl)
Drug and Metabolite Concentrations in Rabbit Serum and Liver MDL numberDiet%serumliver Parent compoundBisQueenParent compoundVisqueen 103,4910.4000000 103,4910.6000000 104,5560.44.9007.900 104,5990.414.80047.200 104,9620.43.8003.600 105,4430.4000000 105,9750.413.80054.600 106,8340.42.9009.100 107,9170.4000000 108,2080.4000000 108,8040.26000000 N = 5 rabbits / group; Starved overnight Rabbits were fed for 7 days (exceptionally, MDL 103,491 was fed for 0.6 days at 0.6%) Diet% = (MDL compound weight / feed weight) × (100) The data in Table 4 were compared to the control values. Serum parent compound = serum parent compound concentration (μg / ml) serum bis (Bis) = serum bisphenol concentration (μg / ml) serum quine = serum diphenoquinone concentration (μg / g) Liver parent compound = liver parent compound concentration (μg / g) Liver bis = liver bisphenol concentration (μg / g) liver quine = liver diphenoquinone concentration (μg / g)
<Example 31>
Determination of Antioxidant Activity and Bioavailability of Compounds of Formula (1) by In Vivo Screening in Male Sprague-Dawley Rats
A. Experimental Protocol
Representative experiments consisted of four to six groups of rats (N = 5 / group), one group being a control group not fed the MDL compound and the other group treated with 0.3% MDL compound. Some of the compounds were repeated at 0.3% or again evaluated at lower doses of 0.1%. 50-100 g of male Sprague-Dawley rats (Harlan Laboratories, Indianapolis, IN) housed in five groups and a dietary mixture containing unlimited water and MDL compound-free or furina rodent animal feed (# 5002) As feed for 4 days. A diet mixture (0.3%) was prepared by mixing 1.2 g of MDL compound with 400 g of Purina rodent animal food (# 5002). The MDL compound was mixed with about 50 g of food using a mortar and pestle. It was added to the remaining feed and mixed for 3 hours on a rotary mixer. On the morning of the fifth day, hungry rats were anesthetized with carbon dioxide and the heart was punctured to collect blood. Rats were sacrificed by cervical dislocation. Body weight and liver weight were reported in g. Feed consumption was recorded as g / day / rat. Dead rats were recorded as mortality. Fresh serum was dispensed and used for clinical chemical analysis, thiobarbituric acid reactive substance (TBRARS) evaluation and conjugated diene determination. Approximately 0.5 ml of serum aliquot and whole liver were frozen at −20 ° C. for subsequent concentration of compounds and metabolites.
B. Clinical Chemistry Analysis
Blood was allowed to coagulate by placing at room temperature for 30 minutes. Serum was obtained after centrifugation at 3000 rpm and 4 ° C. for 10 minutes using a Beckman J-6M / E centrifuge equipped with a JS-4.2 rotor. Fresh serum was prepared from Roche Diagnostic Reagents for clinical and chemical measurements of alkaline phosphatase, alanine transaminase, aspartate aminotransferase, total cholesterol, triglycerides and glucose (each, ALP, kit # 44553; ALT, kit # 42375; AST , Kit # 42381; CHOL, Kit # 44334; TG, Kit # 44120; and GLU, Kit # 44558) were analyzed by COBAS MIRA S automated analyzer (Roche Diagnostics). ALP, ALT and AST were calculated as units / liter. Cholesterol, triglycerides and glucose were calculated as mg / dL.
C. HPLC quantitation of compound and metabolite concentrations in serum and liver
Serum and liver concentrations of the parent compound, metabolite, bisphenol and diphenoquinone were determined by reverse phase HPLC using the Waters 990 Powerline System. A 1 g sample of liver was homogenized with 5.0 ml of PBS at pH 7.4 for 20-30 seconds at control point 5 using a polytron tissue homogenizer. Serum or liver homogenate was extracted as follows: 100 μl of serum or homogenate was added to 2.0 ml of diethyl ether: ethanol (3: 1) while vortexing the tube. The sample tube was capped and centrifuged for 10 minutes at 3500 rpm and 5 ° C. with a Beckman GPKR centrifuge equipped with a GH 3.7 rotor. The supernatant was transferred to a clean tube and dried under nitrogen. Samples were reconstituted with 200 μl of acetonitrile: hexanes: 0.1 M ammonium acetate (90: 6.5: 3.5, volume ratio). 100 μl was then injected onto a Waters Delta Pack C18-300 mm 3 column and eluted at a flow rate of 1.5 ml / min with 83% acetonitrile: 17% water mobile phase. Absorbance was recorded at wavelengths of 240, 254 and 420 nm. Compound concentrations were calculated from known stocks of certain parent compounds after correction for recovery; The recovery range from the spike partial sample ranged from 40 to 100%. The lowest detection limit for this class of compounds was about 0.5 μg / ml. Concentrations were calculated as μg / ml serum and μg / liver g.
D. Evaluation of Thiobarbituric Acid Reactant (TBARS)
In this evaluation, oxidation of serum lipids is initiated with CuSO ₄ , and aldehydes such as malondialdehyde (MDA) are produced. When incubated with thiobarbituric acid, the absorbance of aldehydes can be detected at 530-540 nm. As noted in the previous examples, lower TBARS values than control serum values indicate the relative ability of the test compound to inhibit lipid oxidation of the sample. TBARS was determined as follows: 100 μl of test serum was mixed with 400 μl of 5 mmol CuSO ₄ solution and incubated at 37 ° C. for 3 hours. 1.0 ml of 20% trichloroacetic acid was added to stop the reaction. Then, 1.0 ml of 0.67% thiobarbituric acid in 0.05 N sodium hydroxide was added, mixed, and the samples were incubated at 90 ° C. for 30 minutes. Samples were briefly centrifuged to pellet undissolved material and the supernatant transferred to a 96-well microtiter plate. Absorbance was measured at 540 nm using a Biotek model EL311 microplate reader. The resulting MDA (nmol) was calculated from a standard curve of 0-10 nmol of MDA prepared from malonaldehyde bis (dimethylacetal). Serum samples from treated rats were compared to serum samples from control rats not fed the MDL compound.
E. Conjugated Diene Measurement
The conjugated diene lag phase is another indicator of lipid oxidation. Lipids exposed to Cu ⁺⁺ formed conjugated dienes that absorb ultraviolet light in the range of 230-235 nm. The retarder of diene formation is an indicator of lipid oxidation. Retarders longer than the control sample show inhibition of oxidation. Conjugated diene retarders were measured at 30 ° C. using a Varian DMS200 spectrophotometer (equipped with a constant temperature five cuvette sample exchanger). 20 μl of collected serum was added to a cuvette containing 3.0 ml of phosphate buffered saline at pH 7.5. Absorbance of all cuvettes was measured and the machine baseline was adjusted to zero using the sample with the lowest absorbance. Next, 100 μl of 1 mmol CuSO ₄ was added and mixed immediately. The absorbance of each cuvette was recorded at 2 minute intervals for 840 minutes. Data was obtained and sent to Microsoft® registered trademark EXCEL spreadsheet, with curves flat and different. The retarder was mathematically determined as minutes. Serum samples were collected (N = 5) and the data shown is the average of two measurements. Serum samples from treated rats were compared to serum samples from control rats not fed the MDL compound.
Tables 5, 6 and 7 below summarize the data from the individual experiments of this test method. Table 5 shows serum chemistry in male Sprague-Dawley rats, Table 6 shows animal variables, and Table 7 provides drug or metabolite concentrations in both serum and liver.
Antioxidant Effect of Compounds of Formula (1) in Male Sprague-Dawley Rats as a Percent to Controls MDL numberDiet%ALPASTALTCHOLGLUCTRIGTBARSConjugated diene (min) 103,0760.3201%99%132%114%99%77%107%ND * 103,1410.3147%120%126%119%107%47%113%ND 103,1570.3144%92%133%112%100%82%94%50 103,3770.3147%149%145%93%102%85%71%ND 103,4910.380%114%127%146%94%131%34%ND 103,4910.1116%94%133%112%101%92%70%168 104,3990.391%102%107%130%88%168%41%395 104,5560.3112%101%106%119%113%95%34%ND 104,5560.1112%107%133%96%102%74%61%200 104,5710.3103%103%125%108%96%119%58%ND 104,5990.3110%76%109%94%111%100%34%ND 104,9620.3130%109%91%99%91%74%25%320 105,3140.3154%76%112%110%115%78%85%ND 105,4430.1101%94%113%111%106%111%78%151 105,4430.390%171%156%131%98%126%14%ND 105,7260.3118%112%113%104%105%75%48%ND 105,9750.3105%122%106%122%106%107%24%ND 105,9750.1112%89%96%98%110%108%67%ND 106,2900.3118%84%89%153%109%75%17%372 106,8340.369%122%141%150%95%83%31%492 108,7010.396%96%111%99%112%58%62%274 * ND = not measured N = 5 rats / group diet% = (MDL compound weight / feed weight) × (100) conjugated diene = conjugated diene retarder (min) (average of two measurements of collected samples, N 5) Except for the conjugated diene and diet%, the data in Table 5 was normalized as follows:% Control = (Treatment, Mean / Control, Mean) × (100) ALP = Alkaline Phosphatase, U / mL AST = Aspartate Aminotransferase, U / mL ALT = Alanine Aminotransferase, U / mL CHOL = Total Cholesterol, mg / lTRIG = Triglycerides, mg / l GLUC = Glucose, mg / l TBARS = nmol MDL Thiobarbituric acid reactive substances
Animal Variables as Percent to Controls MDL numberDiet%foodweightlw / bwdeath rate 103,0760.393%105%110%0 % 103,1410.387%91%96%0 % 103,1570.396%96%120%0 % 103,3770.382%97%105%0 % 103,4910.389%100%130%0 % 103,4910.192%95%106%0 % 104,3990.390%101%118%0 % 104,5560.368%87%120%0 % 104,5560.1102%103%106%0 % 104,5710.3108%97%123%0 % 104,5990.3103%93%107%0 % 104,9620.374%95%103%0 % 105,3140.3109%111%118%0 % 105,4430.1100%101%110%0 % 105,4430.384%95%118%0 % 105,7260.3112%105%120%0 % 105,9750.398%100%113%0 % 105,9750.1106%98%105%0 % 106,2900.3108%95%104%0 % 106,8340.391%95%132%0 % 108,7010.390%97%119%0 % N = 5 rats / group diet% = (MDL compound weight / feed weight) × (100) The data in Table 6 was normalized according to the formula shown in Table 5. Feed = amount eaten per day per rat (g) body weight = Weight (g) lw / bw = (liver g / g) mortality = deaths / group
Drug and Metabolite Concentrations in Rat Serum and Liver MDL numberDiet%serumliver Parent compoundVisqueenParent compoundVisqueen 103,0760.3000000 103,1410.3000000 103,1570.3000000 103,3770.37.72 *0022.2 **00 103,4910.3300000 103,4910.1000000 104,3990.3000000 104,5560.316.30060.300 104,5560.19.20038.700 104,5710.31.2000.500 104,5990.328.30044.900 104,9620.316.40043.500 105,3140.30.0000.000 105,4430.1000000 105,4430.30.0 †00000 105,7260.31700128.300 105,9750.335.800185.900 105,9750.123.80094.500 106,2900.34.8 ‡0026.2 ‡‡00 106,8340.31.8002.700 108,7010.380028.200 In addition, 2.3 μg / ml of 2,6-di-t-butyl-4 [(dimethylphenylsilyl) methyloxy] phenol was also observed. ** 19.2 μg / mg of 2,6-di-t-butyl -4 [(dimethylphenylsilyl) methyloxy] phenol was also observed. † 3.8 μg / ml of MDL 103,491 was also observed. ‡ This value represents the amount of MDL 104,962 observed. ‡‡ This value was observed The amount of MDL 104,962 is shown. The data in Table 7 is shown as mean (N = 5) and not normalized to the control values. Serum parent compound = serum parent compound concentration (μg / ml) Serum bis = Serum bisphenol concentration (μg / ml) serum quin (serum) = serum diphenoquinone concentration (μg / g) liver parent compound = liver parent compound concentration (μg / g) liver bis = liver bisphenol concentration (μg / g) Liver Quine = Liver Diphenoquinone Concentration (μg / g)
<Example 32>
Inhibitory Effect of Compounds of Formula (1) on Atherosclerosis in Cholesterol-fed Female New Zealand White Rabbits
A. Experimental Protocol
Four independent experiments were performed. Each experiment consisted of 1 to 5 groups (N = 5 rats / group) treated with control and MDL compounds. Female New Zealand white rabbits (Hazelton, about 2.0-2.3 kg) were fed 1% cholesterol supplemented rabbit food (Purina # 5322) with or without 0.4% MDL compound. The MDL compound was dissolved in 100% ethanol, sprayed on the food and dried overnight in a chemical vapor hood. Alternatively, the MDL compound can be mixed in furina rabbit food. Ethanol was injected into the control food. The rabbits were fed 100 g of food per day for 70 days, with no water restrictions. About 2 ml of blood (overnight) rabbits were drawn from the marginal ear vein and serum cholesterol levels were monitored regularly. Rabbits were euthanized at 70 days with carbon dioxide overdose. Body weight and liver weight were reported in grams. Feed consumption was recorded in g / day. Fresh serum was dispensed and used for clinical chemical analysis, lipoprotein cholesterol determination, thiobarbituric acid reactive substance (TBRARS) evaluation, and determination of compound and metabolite concentrations in serum. The liver (approximately 5 g aliquots) was frozen at −20 ° C. for subsequent determination of compound and metabolite concentrations.
The aorta was dissected immediately after the rabbits were sacrificed. After debridement of the foreign adipose tissue, the aorta was excised from the elevation artery for the iliac branch. The aorta was stored overnight in phosphate buffered saline at pH 7.4 at 4 ° C. until final debridement. Arteries were longitudinally cut open and stained with Sudan IV. After staining, a pin was inserted into the aorta to be flattened, and an area of the proteolytic lesion was quantified after obtaining an electronic image.
B. Clinical Chemistry Analysis
Blood was allowed to coagulate by placing at room temperature for 30 minutes. Serum was obtained after centrifugation at 3000 rpm and 5 ° C. for 10 minutes using a Beckman GPKR centrifuge equipped with a GH 3.7 rotor. Fresh serum was analyzed with a COBAS MIRA S Automated Analyzer (Roche Diagnostics) using Roche Diagnostic Reagents for Total Cholesterol and Triglycerides (CHOL, Kit # 44334; and TG, Kit # 44120, respectively). Cholesterol and triglycerides were calculated as mg / dl.
C. TBARS Assessment
Oxidation of serum lipids is initiated with CuSO ₄ to produce aldehydes such as malondialdehyde (MDA). When incubated with thiobarbituric acid, the absorbance of aldehydes can be detected at 530-540 nm. TBARS was determined as follows: 50 μl of serum was mixed with 50 μl of 0.9% physiological saline and 400 μl of 5 mmol CuSO ₄ solution and incubated at 37 ° C. for 5 hours. 1.0 ml of 20% trichloroacetic acid was added to stop the reaction. Then, 1.0 ml of 0.67% thiobarbituric acid in 0.05 N sodium hydroxide was added, mixed, and the samples were incubated at 90 ° C. for 30 minutes. Samples were briefly centrifuged to pellet undissolved material and the supernatant transferred to a 96-well microtiter plate. Absorbance was measured at 540 nm using a Biotek model EL311 microplate reader. The resulting MDA (nmol) was calculated from a standard curve of 0-10 nmol of MDA prepared from malonaldehyde bis (dimethylacetal). Serum samples from treated rabbits were compared to serum samples from control rabbits not fed the MDL compound.
D. HPLC quantitation of compound and metabolite concentrations in serum and liver
Serum and liver concentrations of the parent compound, metabolite, bisphenol and diphenoquinone were determined by reverse phase HPLC using the Waters 990 Powerline System. 1 g of liver was homogenized with 5.0 ml of PBS at pH 7.4 for 20-30 seconds at control point 5 using a polytron tissue homogenizer. Serum or liver homogenate was extracted as follows: 100 μl of serum or homogenate was added to 2.0 ml of diethyl ether: ethanol (3: 1) while vortexing the tube. The sample tube was capped and centrifuged for 10 minutes at 3500 rpm and 5 ° C. with a Beckman GPKR centrifuge equipped with a GH 3.7 rotor. The supernatant was transferred to a clean tube and dried under nitrogen. Samples were reconstituted with 200 μl of acetonitrile: hexanes: 0.1 M ammonium acetate (90: 6.5: 3.5, volume ratio). 100 μl was then injected onto a Waters Delta Pack C18-300 mm 3 column and eluted at a flow rate of 1.5 ml / min with 83% acetonitrile: 17% water mobile phase. Absorbance was recorded at wavelengths of 240, 254 and 420 nm. Compound concentrations were calculated from known amounts of the parent compound after correcting for recovery. Concentrations were calculated as μg / ml serum and μg / liver g.
E. HPLC Separation and Quantitative Analysis of Lipoprotein Small Fraction Cholesterol Levels
Lipoprotein fractions of VLDL, LDL and HDL were separated on a Sepharose 6HR column (1 × 30 cm, Pharmacia) connected to a Waters Powerline HPLC system. 50 μl of serum was injected onto the column and eluted at a flow rate of 0.5 ml / min with phosphate buffered saline at pH 7.4. Cholesterol reagent (Roche Diagnostics, Kit # 44334, diluted with 20 ml of water, then diluted with 20 ml of 0.9% saline solution) was added to the next column eluent at a flow rate of 0.2 ml / min, and a knitted PFTE Kratos reaction coil (Applied Biosystems Incubated at 37 ° C. for 5 minutes. Absorbance was measured at 500 nm. Lipoprotein subfractions were quantified as follows: (total serum cholesterol) × (% area under the curve for each subfraction).
In addition, the compounds of the formula (1) may be used as chemical antioxidants, for example, in organic materials such as rubber, plastics, fats, petroleum products, etc. which are usually subjected to oxidative degradation. In general, the preservative effective amount of the compound of formula (1) is a concentration sufficient to inhibit oxidative degradation of the substance to be protected. The preservative effective amount of formula (1) is generally varied from about 0.01% to about 1.0% by weight.

权利要求:
Claims (43)
[1" claim-type="Currently amended] Compound of formula (1) or a pharmaceutically acceptable salt thereof.
<Formula 1>

In the above formula,
R ₁ and R ₆ are each independently C ₁ -C ₆ alkyl;
R ₂ , R ₃ and R ₄ are each independently hydrogen or C ₁ -C ₆ alkyl;
R is hydrogen or —C (O) — (CH ₂ ) _m -Q wherein Q is hydrogen or —COOH and m is an integer of 1, 2, 3 or 4;
Z is a thio, oxy or methylene group;
A is a C ₁ -C ₄ alkylene group;
R ₅ and R ₇ are each independently C ₁ -C ₆ alkyl or — (CH ₂ ) _n — (Ar), where n is an integer 0, 1, 2 or 3; Ar is hydroxy, methoxy, 1 to 1 selected from the group consisting of oxy, halogen, trifluoromethyl, C ₁ -C ₆ alkyl, or —NR ₈ R ₉ , wherein R ₈ and R ₉ are each independently hydrogen or C ₁ -C ₆ alkyl. Phenyl or naphthyl unsubstituted or substituted with three substituents; Provided that when at least one of R ₂ and R ₅ or R ₇ is C ₁ -C ₆ alkyl and Ar is not substituted with trifluoromethyl or —NR ₈ R ₉ , then R is —C (O) — (CH ₂ ) _m -Q.
[2" claim-type="Currently amended] The compound of claim 1, wherein R ₁ is methyl or tertiary butyl; R ₂ and R ₃ are each independently hydrogen, methyl or tertiary butyl; R ₄ is hydrogen or methyl; R ₅ is methyl; R ₆ is methyl or phenyl; R is hydrogen, acetyl or succinyl.
[3" claim-type="Currently amended] The compound of claim 1, wherein R ₇ is — (CH ₂ ) _n — (Ar) where n is an integer 0, 1, 2 or 3; Ar is phenyl substituted with 1-3 substituents of —NR ₈ R _9. ) Compound.
[4" claim-type="Currently amended] The compound of claim 3, wherein R is hydrogen, acetyl or succinyl; R ₁ is methyl or tertiary butyl; R ₂ and R ₃ are each independently hydrogen, methyl or tertiary butyl; R ₄ is hydrogen or methyl; R ₅ and R ₆ are each methyl.
[5" claim-type="Currently amended] The compound of claim 4, wherein R ₈ and R ₉ are each methyl and R is hydrogen.
[6" claim-type="Currently amended] The compound of claim 1, wherein R is —C (O) — (CH ₂ ) _m -Q, wherein Q is hydrogen or —COOH and m is an integer of 1, 2, 3, or 4. 8.
[7" claim-type="Currently amended] The compound of claim 6, wherein R ₁ is methyl or tertiary butyl; R ₂ and R ₃ are each independently hydrogen, methyl or tertiary butyl; R ₄ is hydrogen or methyl; R ₅ is methyl; R ₆ is methyl or phenyl; R ₈ and R ₉ are each methyl.
[8" claim-type="Currently amended] The compound of claim 2, wherein Z is thio.
[9" claim-type="Currently amended] The compound of claim 2, wherein Z is oxy.
[10" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,6-di-t-butyl-4-[(diphenylmethylsilyl) methyloxy] phenol.
[11" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,6-di-t-butyl-4-[(4-N, N-dimethylaminophenyldimethylsilyl) methyloxy] phenol.
[12" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-4-trifluoromethylphenylsilyl) methyloxy] phenol.
[13" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,6-di-t-butyl-4-[(dimethyl-3-trifluoromethylphenylsilyl) methyloxy] phenol.
[14" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2-t-butyl-4-[(dimethylphenolsilyl) methyloxy] phenol.
[15" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol succinic acid ester.
[16" claim-type="Currently amended] The compound of claim 1, wherein said compound is 2-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol succinic acid ester.
[17" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,6-di-t-butyl-4-[(dimethylphenylsilyl) methylthio] phenol succinic acid ester.
[18" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,6-di-t-butyl-4-[(trimethylsilyl) methylthio] phenol succinic acid ester.
[19" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol acetic acid ester.
[20" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,6-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol acetic acid ester.
[21" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,3,6-trimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol acetic acid ester.
[22" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,5-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol.
[23" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,5-di-t-butyl-4-[(dimethylphenylsilyl) methyloxy] phenol acetic acid ester.
[24" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2-t-butyl-4-[(dimethylphenylsilyl) methylthio] phenol.
[25" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,3,6-trimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol.
[26" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,3,5-trimethyl-4-[(dimethylphenylsilyl) methyloxy] phenol.
[27" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2-t-butyl-4-[(dimethyl-p-methoxyphenylsilyl) methyloxy] phenol.
[28" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,5-di-t-butyl-4-[(diphenylmethylsilyl) methyloxy] phenol.
[29" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,6-di-t-butyl-4-[(diphenylmethylsilyl) methyloxy] phenol.
[30" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2,6-di-t-butyl-4-[(methyl-di-p-methoxyphenylsilyl) methyloxy] phenol.
[31" claim-type="Currently amended] The compound of claim 1, wherein the compound is 2-t-butyl-4-[(dimethylbenzylsilyl) methyloxy] phenol.
[32" claim-type="Currently amended] A method of inhibiting the progression of atherosclerosis in a patient comprising administering to the patient in need of inhibiting the progression of atherosclerosis an effective amount of the compound atherosclerosis inhibitory effect of claim 1.
[33" claim-type="Currently amended] A method for treating atherosclerosis in a patient comprising administering to said atherosclerosis patient an effective amount of the compound atherosclerosis inhibiting compound of claim 1.
[34" claim-type="Currently amended] A method of inhibiting peroxidation of LDL cholesterol in a patient comprising administering an effective amount of the compound antioxidant of claim 1 to a patient in need of inhibiting the peroxidation of LDL cholesterol.
[35" claim-type="Currently amended] A method of lowering plasma cholesterol levels in a patient comprising administering to the patient in need thereof a plasma cholesterol level lowering compound of claim 1.
[36" claim-type="Currently amended] In said patient comprising administering to said patient in need of inhibiting cytokine-induced expression of vascular cell adsorption molecule-1 and / or intercellular adsorption molecule-1 said compound expression inhibitory effective amount of said compound of claim 1. A method of inhibiting cytokine-induced expression of.
[37" claim-type="Currently amended] A method of treating a patient, comprising administering to said patient a chronic inflammatory disease a therapeutically effective amount of the compound of claim 1.
[38" claim-type="Currently amended] The method of claim 37, wherein the inflammatory disease is asthma.
[39" claim-type="Currently amended] The method of claim 37, wherein the inflammatory disease is chronic inflammation.
[40" claim-type="Currently amended] The method of claim 37, wherein the inflammatory disease is rheumatoid arthritis.
[41" claim-type="Currently amended] The method of claim 37, wherein the inflammatory disease is autoimmune diabetes.
[42" claim-type="Currently amended] The method of claim 37, wherein the inflammatory disease is transplant rejection.
[43" claim-type="Currently amended] The method of claim 37, wherein the inflammatory disease is tumor angiogenesis.

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CA2251991C|2002-05-14|

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EP0900225B1|2002-01-30|

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WO1997041129A1|1997-11-06|

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AU1985497A|1997-11-19|

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PT900225E|2002-06-28|

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NO985039L|1998-12-21|

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JP2001509782A|2001-07-24|

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ES2167716T3|2002-05-16|

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DK0900225T3|2002-05-13|

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KR100503181B1|2006-03-03|

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CA2251991A1|1997-11-06|

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CN1216993A|1999-05-19|

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AR006890A1|1999-09-29|

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BR9709190A|1999-08-10|

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AT212639T|2002-02-15|

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ZA9703503B|1997-10-30|

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NO985039D0|1998-10-29|

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DE69710191D1|2002-03-14|

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引用文献:

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公开号 | 申请日 | 公开日 | 申请人 | 专利标题

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法律状态:
1996-04-30|Priority to US8/637,968

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1996-04-30|Priority to US08/637,968

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1996-04-30|Priority to US08/637,968

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1997-03-03|Application filed by 게리 디. 스트리트, 스티븐 엘. 네스비트, 훽스트 마리온 로우셀, 인크.

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1997-03-03|Priority to PCT/US1997/003335

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2000-11-06|Publication of KR20000065101A

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2006-03-03|Application granted

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2006-03-03|Publication of KR100503181B1

优先权:

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申请号 | 申请日 | 专利标题

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US8/637,968|1996-04-30|

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US08/637,968|1996-04-30|

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US08/637,968|US5608095A|1996-04-30|1996-04-30|Alkyl-4-silyl-phenols and esters thereof as antiatherosclerotic agents|

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PCT/US1997/003335|WO1997041129A1|1996-04-30|1997-03-03|Alkyl-4-silyl-phenols and esters thereof as antiatherosclerotic agents|