Compounds of cyclic phenolic thioethers which are useful in stimulating and inhibiting superoxide generation

The present invention relates to compounds of the formula: ##STR1## wherein R.sup.1 and R.sup.2 are the same or different and independently represent tert-alkyl, phenyl, or hydrogen; R.sup.3 represents hydrogen or alkyl; X represents O, S or (CH.sub.2).sub.m wherein m is 1 or 2; A represents O or S(O).sub.n wherein n is 0, 1, or 2; p is an integer from 0 to 4; and R represents alkyl; OH; OR.sup.4 wherein R.sup.4 is alkyl; or NR.sup.5 R.sup.6 wherein R.sup.5 is hydrogen or alkyl, and R.sup.6 is hydrogen, alkyl, heterocyclealkyl, substituted heterocyclealkyl, cycloalkyl, substituted cycloalkyl, phenyl, substituted phenyl, phenylalkyl, or substituted phenylalkyl, or NR.sup.5 R.sup.6 together form a heterocyclic ring which may optionally be substituted; or (CH.sub.2).sub.t COOR.sup.7 wherein t is an integer from 1 to 4 and R.sup.7 is hydrogen or alkyl; and the pharmaceutically acceptable salts thereof. The compounds are inhibitors or stimulators of superoxide generation.

BACKGROUND OF THE INVENTION 
1. Field of the Invention 
The present invention relates to cyclic phenolic thioethers and more 
particularly relates to the novel compounds of Formula I which are 
inhibitors or stimulators of superoxide generation and may also inhibit 
cyclooxygenase or 5-lipoxygenase. The compounds of the present invention 
which stimulate superoxide generation may be useful as adjunctive 
therapeutic agents in the treatment of infections. Other compounds of the 
present invention which inhibit superoxide generation may be useful in the 
therapeutic or prophylactic treatment of disease conditions which are 
mediated wholly or partly by superoxide generation such as adult 
respiratory distress syndrome, superoxide mediated inflammatory or 
allergic conditions, and other medical conditions which are caused by or 
aggravated by superoxide. 
The compounds of Formula I which inhibit cyclooxygenase or 5-lipoxygenase 
are useful, for example, as anti-inflammatory and/or anti-allergy agents 
and in the treatment of hypersensitivity reactions, psoriasis, asthma, and 
related disorders and conditions in which physiologically active agents 
formed in the arachidonic acid metabolic pathway are involved. Compounds 
of the present invention may be useful in treating inflammatory and 
allergic conditions such as arthritis, asthma, and psoriasis. 
2. Background Information 
Recently, oxygen radicals have been implicated in the pathogenesis of many 
diseases. This implication is reflected by the many conferences devoted to 
this topic, books on the subject of free radicals and disease, and the 
appearance of two new specialized journals: Free Radical Research 
Communications, and Free Radical Biology and Medicine. 
Much is known about the physicochemical properties of the various oxygen 
radicals, but knowledge of their overall importance in the initiation and 
amplification of human disease is limited. Some clinical conditions in 
which oxygen radicals are thought to be involved are discussed in Cross, 
C. E., et al., "Oxygen Radicals and Human Disease," ANN. INT. MED., 
107:526-545 (1987) (see Table 1, p. 527) and Ward, P. A., et al., "Oxygen 
Radicals, Inflammation, and Tissue Injury," FREE RADICAL BIOLOGY & 
MEDICINE, 5:403-408 (1988). Among the clinical conditions in which oxygen 
radicals are thought to be involved are, for example, inflammatory-immune 
injury, autoimmune diseases, ischemia-reflow states, aging disorders, 
cancer, cigarette-smoke effects, emphysema, acute respiratory distress 
syndrome (ARDS), atherosclerosis, rheumatoid arthritis, senile dementia, 
cataractogenesis, retinopathy of prematurity, radiation injury and contact 
dermatitis. 
Oxygen radicals are capable of reversibly or irreversibly damaging 
compounds of all biochemical classes, including nucleic acids, protein and 
free amino acids, lipids and lipoproteins, carbohydrates, and connective 
tissue macromolecules. These species may have an impact on such cell 
activities as membrane function, metabolism, and gene expression. Oxygen 
radicals are formed in tissues by many processes (see Cross, et al., p. 
528, Table 2). These are believed to be both endogenous, such as 
mitochondrial, microsomal and chloroplast electron transport chains; 
oxidant enzymes such as xanthine oxidase, indoleamine dioxygenase, 
tryptophan dioxygenase, galactose oxidase, cyclooxygenase, lipoxygenase, 
and monoamine oxidase; phagocytic cells such as neutrophils, monocytes and 
macrophages, eosinophils, and endothelial cells; and antioxidation 
reactions; and exogenous, such as redox-cycling substances, drug 
oxidations, cigarette smoke, ionizing radiation, sunlight, heat shock and 
substances that oxidize glutathione. They may be involved in the action of 
toxins such as paraquat, cigarette smoke, and quinone antitumor drugs. 
Those compounds of the present invention which inhibit superoxide 
generation may be useful in the treatment of diseases mediated by 
superoxide generation. 
There are also some conditions in which the generation of superoxide may be 
desirable. Those compounds of the present invention which stimulate 
superoxide generation may be useful in the adjunctive therapy of microbial 
infections. See Goodman and Gilman's, The Pharmacological Basis of 
Therapeutics (7th Edition, 1985) p. 660-673; P. A. Ward, et. al., "Oxygen 
Radicals, Inflammation and Tissue Injury," FREE RADICAL BIOLOGY & 
MEDICINE, 5: 403-408 (1988); and C. E. Cross, et. al., "Oxygen Radicals 
and Human Disease,"; ANN. INT. MED., 107: 526-545 (1987). Generation of 
reactive oxygen species is a critical event in successful host defense 
against invading organisms. Both neutrophils and macrophages rely on a 
variety of oxidants to damage bacterial constituents (see V. L. Shepherd, 
"The role of the respiratory burst of phagocytes in host defense," SEMIN. 
RESPIR. INFECT. (United States) Jun. 1986, 1(2) p. 99-106. 
It is well recognized that arachidonic acid, an essential unsaturated fatty 
acid, is enzymatically oxygenated to various products, including, 
prostaglandins, thromboxanes, the 5-, 11-, 12- and 
15-hydroxyeicosatetraenoic acids (HETEs, DIHETEs) and 
hydroperoxyeicosatetraenoic acids (HPETEs) and the leukotrienes, all of 
which have potent physiological effects. 
Those compounds of the present invention which inhibit cyclooxygenase 
inhibit the synthesis of prostaglandins via the cyclooxygenase pathway of 
arachidonic acid metabolism. These prostaglandin synthetase inhibitors may 
exhibit anti-inflammatory, anti-pyretic and analgesic activity, and are 
useful in the treatment of inflammatory conditions such as arthritis. 
The leukotrienes, which are produced via the 5-lipoxygenase pathway, are 
the major contributors to the onset of the symptoms of asthma, and 
mediators for immediate hypersensitivity reactions, inflammation and other 
allergic responses. 
Leukotrienes are found in inflammatory exudates and are involved in the 
process of cellular invasion during inflammation. The term "leukotrienes" 
is used as a generic term to describe a class of substances, such as 
slow-reacting substance (SRS) which is an important mediator in asthma and 
other hypersensitivity reactions. Immunologically generated SRS is usually 
referred to as slow-reacting substance of anaphylaxis (SRS-A). SRS-A 
consists of leukotrienes (LT) known as A.sub.4, B.sub.4, C.sub.4, D.sub.4, 
and E.sub.4. LTC.sub.4 is at least 100 times more potent than histamine in 
causing long lasting bronchoconstricting effects. The leukotrienes also 
increase vascular permeability and cause decreased cardiac output and 
impaired ventricular contraction. LTB.sub.4 may be an important mediator 
of inflammation in, for example, inflammatory bowel disease. 
Chemotaxis is a reaction by which the direction of migration of cells is 
determined by substances in their environment. It is one of the major 
processes bringing leukocytes from the blood to an inflammatory site, 
whether the inflammation is caused by an infectious agent, allergic 
challenge, or other pro-inflammatory stimuli. LTB.sub.4 is not only 
chemotactic for neutrophils and monocytes, but is also highly active in 
stimulating eosinophil locomotion. LTB.sub.4 also stimulates calcium 
influx and aggregation of polymorphonuclear leukocytes and LTB.sub.4 may, 
thus, play an important role in mediating both acute and chronic 
inflammation. 
Rheumatoid spondylitis is characterized by an acute neutrophil flareup in 
the joint which is associated with elevated levels of LTB.sub.4. LTB.sub.4 
is also present in gouty effusions; and exposure to urate crystals is 
known to stimulate LTB.sub.4 production by neutrophils. Accordingly, those 
compounds of the present invention which inhibit 5-lipoxygenase through 
inhibition of neutrophil attraction and activation in arthritic joints 
should reduce the protease and oxidative burden believed responsible for 
joint destruction in arthritic diseases. 
Prior to the recognition of the significance of the arachidonic acid 
metabolism pathway in allergic reactions and inflammation, the search for 
effective therapeutic agents was based primarily on those agents which 
treated the symptoms of allergy and inflammation. There has since been an 
effort to develop new drugs which selectively block the formation of the 
mediators of these conditions, and the present invention provides new 
chemical entities which are inhibitors of the arachidonic acid pathway and 
are useful in the treatment of asthma, rheumatoid arthritis, psoriasis, 
and other allergic, hypersensitivity, and inflammatory conditions. 
Various thioether compounds have been described previously. For example, 
U.S. Pat. No. 4,711,903 and its continuation-in-part, 4,755,524 disclose 
compounds of the formula 
##STR2## 
wherein: R.sub.1 and R.sub.2 are the same or different and independently 
represent tert-alkyl or phenyl; A represents methylene or methylene 
substituted by alkyl, dialkyl or hydroxy, provided that when A includes 
hydroxymethylene, the hydroxymethylene group is not adjacent to a 
heteroatom; B represents sulfur, sulfoxide, sulfone, oxygen, --NH-- or 
nitrogen substituted by alkyl, phenyl, benzyl, substituted phenyl or 
substituted benzyl; C represents methylene or methylene substituted by 
alkyl; R.sub.3 represents CO.sub.2 H, CO.sub.2 -alkyl or a tetrazole 
group; m is 0 or 1, n is 2, 3 or 4 and p is 1, 2 or 3; and the 
pharmaceutically acceptable salts thereof. The compounds are specific 
inhibitors of 5-lipoxygenase and are useful in the treatment of local and 
systematic inflammation, allergy and hypersensitivity reactions and other 
disorders in which agents formed in the 5-lipoxygenase metabolic pathway 
are involved. 
U.S. Pat. No. 4,621,098 and its equivalent, European Patent Application 
Publication No. 0131221 discloses compounds of the formula 
##STR3## 
in which Ar is phenyl or phenyl substituted by one to three of varied 
substituents, for example, alkyl, alkoxy, hydroxy, etc.; Q is oxygen, 
sulfur or an NH group; A is straight or branched chain, optionally 
substituted, alkylene, and R is hydrogen or straight or branched alkyl, 
optionally substituted by alkoxy, hydroxyl, carboxyl, alkoxycarbonyl, 
etc.; and n is 0, 1 or 2. The disclosed compounds are indicated to have 
anti-inflammatory and anti-allergic properties through inhibition of 
undefined anaphylactic and anaphylactoid reactions, although no test data 
are provided. The preferred compounds are stated to be those in which Q 
represents oxygen and n is 0 without mention of any preference among the 
numerous possible substituents for R or substituted phenyl as Ar. In 
contrast to the invention disclosed in the foregoing publication, the 
compounds of the present invention all have cycloalkyl at the position 
corresponding to A as well as having di(tertiary)-alkyl or diphenyl groups 
as substituents on the phenol moiety corresponding to the substituted Ar 
group in the above publication which, as described therein, may or may not 
comprise a phenol. 
U.S. Pat. Nos. 4,029,812, 4,076,841 and 4,078,084 disclose compounds of the 
formula 
##STR4## 
comprising 2-(3,5-di-tert -butyl-4-hydroxy-phenyl) thio carboxamides. The 
compounds are indicated to be useful in lowering serum cholesterol and 
triglyceride levels. 
A series of thioethers, useful as, for example, polyfunctional antioxidants 
for polymers, and biologically active substances, obtained by the 
nucleophilic addition of thiols, including 
3,5-di-tertbutyl-4-hydroxythio-phenol, and hydrogen sulfide to acrylate 
derivatives have been described. See Medvedev et al., Khimiya; 
Khimicheskaya Tekhnologiya, Volume 20, (1977), pp. 568-574. The compounds 
resulting from the foregoing process have the general formulas 
RS(CH.sub.2).sub.n X and S(CH.sub.2 CH.sub.2 X).sub.2 in which R is 
3,5-di-tert-butyl-4-hydroxyphenyl and X represents, for example, 
--C.tbd.N, NH.sub.2, CH(OH)CH.sub.2 Cl, OH, COCl and various carboxy, 
carboxylate and amide functions. Compounds of formula I according to the 
present invention or 5-lipoxygenase activity for structurally related 
compounds are not disclosed. 
U.S. Pat. No. 4,153,803 discloses cholesterol-lowering phenoxyalkanoic acid 
esters of the formula 
##STR5## 
wherein, when Y is sulfur, X is hydrogen, benzyl, benzyloxy or benzylthio 
or substituted derivatives thereof; R is hydrogen, halogen, hydroxy, alkyl 
or alkoxy, A.sup.1 and A.sup.2 are hydrogen or alkyl and Z is amine or 
azacyclohydrocarbonyloxy. 
JP 49116035 discloses a process for making compounds of the formula 
##STR6## 
wherein R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are hydrogen, alkyl or aryl 
groups, and R.sup.1 and R.sup.2 can be combined to form a cycloalkyl 
group. The compounds are said to be useful as drug intermediates, 
agricultural chemicals, antioxidants and industrial chemicals. 
Specifically disclosed is a compound of the formula 
##STR7## 
SUMMARY OF THE INVENTION 
It is, therefore, a primary object of the present invention to provide 
novel cyclic phenolic thioethers pharmaceutical compositions containing 
them and methods of using them, as well as intermediates for producing 
them. 
It is a further object of the present invention to provide methods for 
stimulating or inhibiting superoxide generation and to provide methods for 
treating conditions mediated by products of the arachidonic acid metabolic 
pathway and for promoting anti-inflammatory and/or anti-allergic effects 
in mammals in need thereof by the administration of preselected dosages of 
compounds of the present invention or pharmaceutically acceptable salts 
thereof in appropriate non-toxic pharmaceutical dosage forms or 
compositions. 
Another object of the present invention is to provide dosage unit forms 
adapted for, e.g., oral, topical, and/or parenteral administration and 
useful in the stimulation or inhibition of superoxide generation and in 
the treatment, management and mitigation of inflammation, allergies, 
psoriasis and hypersensitivity reactions and related disorders and 
conditions in which physiologically active agents formed in the 
arachidonic acid metabolic pathway are involved. 
Those compounds of the present invention which inhibit superoxide 
generation may be useful in the therapeutic or prophylactic treatment of 
disease conditions which are mediated wholly or partly by superoxide 
generation such as adult respiratory distress syndrome, superoxide 
mediated inflammatory or allergic conditions, and other medical conditions 
which are caused by or aggravated by superoxide. 
Those compounds of Formula I which are stimulators of superoxide generation 
in neutrophils may be useful in the therapeutic or prophylactic treatment 
of disease conditions in which superoxide generation is an important 
factor. 
Although it has been speculated that 5-lipoxygenase may be involved in 
superoxide generation, the ability of some compounds, which inhibit 
5-lipoxygenase, to stimulate superoxide generation in neutrophils while 
others inhibit superoxide generation indicates that superoxide generation 
is not governed by 5-lipoxygenase. Thus the activity of the compounds of 
Formula I in stimulating or inhibiting superoxide generation is not 
related to the ability to inhibit 5-lipoxygenase. Compounds which do not 
inhibit 5-lipoxygenase may still act as inhibitors or stimulators of 
superoxide generation. In general those compounds of Formula I which are 
carboxylic acids inhibit superoxide generation and those compounds which 
are esters or heterocycle alkyl amides stimulate superoxide generation. 
Compounds of Formula I which are readily hydrolyzable to the carboxylic 
acid upon oral administration may also act as prodrugs which would be 
converted to superoxide inhibitors by stomach acid, blood, liver, or other 
organs. 
In general, those compounds of Formula I wherein R.sup.1 and R.sup.2 are 
tert-alkyl or phenyl and A is sulfur are inhibitors of 5-lipoxygenase. 
The present invention provides a method by which neutrophil activation and 
the generation of superoxide anions are accomplished utilizing compounds 
of Formula I having the ability to stimulate superoxide generation. 
Accordingly these compounds of Formula I are useful in the design and 
testing of anti-inflammatory properties of other pharmacologically active 
agents. 
The ability to produce superoxide which may itself be microbicidal or which 
is then converted to toxic oxidants such as H.sub.2 O.sub.2, OH, and 
singlet oxygen is important to the phagocytic killing mechanisms which 
enable neutrophils and macrophages to kill bacteria and parasites through 
phagocytosis. 
Therefore, compounds of Formula I which stimulate superoxide generation may 
be useful in the adjunctive therapy of microbial infections. The compounds 
may also be useful in treating conditions such as Chediak-Higashi Syndrome 
in which the patient's macrophages and polymorphs are only weakly active 
causing the patients to suffer from recurring infections involving 
organisms with normally low pathogenicity. Compounds of Formula I may also 
be useful in the adjunctive therapy of patients whose immune systems have 
been weakened or impaired by disease or by chemotherapy or radiation 
therapy and who are more subject to microbial infections.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS 
These and other similar objects, advantages and features are accomplished 
according to the products, compositions and methods of the invention 
comprising compounds of the formula 
##STR8## 
and the pharmaceutically acceptable salts thereof wherein R.sup.1 and 
R.sup.2 are the same or different and independently represent tert-alkyl 
of 4 to 10 carbon atoms, phenyl, or hydrogen; R.sup.3 represents hydrogen 
or alkyl of 1 to 4 carbon atoms; X represents O, S or (CH.sub.2).sub.m 
wherein m is 1 or 2; A represents O or S(O).sub.n wherein n is 0, 1, or 2; 
p is an integer from 0 to 4; and R represents: 
(a) alkyl of 1 to 4 carbon atoms; 
(b) OH; 
(c) OR.sup.4 wherein R.sup.4 is alkyl of 1 to 4 carbon atoms; 
(d) NR.sup.5 R.sup.6 wherein R.sup.5 is hydrogen or alkyl, and R.sup.6 is 
hydrogen, alkyl, heterocyclealkyl, substituted heterocyclealkyl, 
cycloalkyl, substituted cycloalkyl, phenyl, substituted phenyl, 
phenylalkyl, or substituted phenylalkyl, or NR.sup.5 R.sup.6 together form 
a heterocyclic ring which may optionally be substituted; or 
(e) (CH.sub.2).sub.t COOR.sup.7 wherein t is an integer from 1 to 4 and 
R.sup.7 is hydrogen or alkyl of 1 to 4 carbon atoms. 
Included in the present invention are compounds of the formula 
##STR9## 
and the pharmaceutically acceptable salts thereof wherein R.sup.1 and 
R.sup.2 are the same or different and independently represent tert-butyl, 
phenyl, or hydrogen with the proviso that when R.sup.1 is hydrogen, 
R.sup.2 is tert-butyl or phenyl; R.sup.3 represents hydrogen or alkyl of 1 
to 4 carbon atoms; X represents (CH.sub.2).sub.m wherein m is 1 or 2; A 
represents O or S; and R represents: 
(a) OH; 
(b) OR.sup.4 wherein R.sup.4 is alkyl of 1 to 4 carbon atoms; or 
(c) NR.sup.5 R.sup.6 wherein R.sup.5 is hydrogen or alkyl of 1 to 4 carbon 
atoms and R.sup.6 is hydrogen, alkyl of 1 to 4 carbon atoms or 
heterocyclealkyl wherein the alkyl moiety has 1 to 4 carbon atoms; or 
NR.sup.5 R.sup.6 together form a heterocyclic ring which may optionally be 
substituted. 
Also included in the present invention are novel intermediates of the 
Formula III 
##STR10## 
wherein R.sup.1, R.sup.2, R.sup.3 and X, are defined as in Formula I, and 
Z represents hydroxy, halogen, sulfate ester, or perfluoroacyl ester. 
The compounds of Formula III are useful as intermediates for making 
compounds of Formula I. 
More preferred compounds of the present invention are compounds of the 
formula IV: 
##STR11## 
wherein R.sup.1 and R.sup.2 are the same or different and independently 
represent tert-butyl, phenyl or hydrogen with the proviso that when 
R.sup.1 is hydrogen, R.sup.2 is tert-butyl or phenyl; X is 
(CH.sub.2).sub.m wherein m is 2, A is S or O; and R is: 
(a) OH; 
(b) OR.sup.4 wherein R.sup.4 is alkyl of 1 to 4 carbon atoms; or 
(c) NR.sup.5 R.sup.6 wherein R.sup.5 is alkyl of 1 to 4 carbon atoms and 
R.sup.6 is alkyl of 1 to 4 carbon atoms or heterocyclealkyl wherein the 
alkyl moiety has 1 to 4 carbon atoms 
and the pharmaceutically acceptable salts thereof. 
Particularly preferred compounds of formula I are those wherein R.sup.1 and 
R.sup.2 are both tert-butyl; X is (CH.sub.2).sub.m wherein m is 2; A is S 
or O; and R is: 
(a) OH; 
(b) OR.sup.4 wherein R.sup.4 is alkyl of 1 to 2 carbon atoms; or 
(c) NR.sup.5 R.sup.6 wherein R.sup.5 alkyl of 1 to 4 carbon atoms and 
R.sup.6 is heterocyclealkyl wherein the alkyl moiety has 1 to 4 carbon 
atoms; 
and the pharmaceutically acceptable salts thereof. 
Compounds of Formula I can possess one or more asymmetric atoms and are 
thus capable of existing in the form of optical isomers as well as in the 
form of racemic or nonracemic mixtures thereof. The optical isomers can be 
obtained by resolution of the racemic mixtures by conventional processes. 
Included in the family of compounds of Formula I and Formula III are all 
isomeric forms thereof, including diastereoisomers, geometric isomers, and 
the pharmaceutically acceptable salts thereof. 
The term "tert-alkyl" as used herein in reference to R.sup.1 and R.sup.2 
refers to branched chain alkyl moieties of from about 4 to about 10 carbon 
atoms having a tertiary carbon atom attached to the phenyl ring 
substituted by R.sup.1 and R.sup.2. Examples of such groups are 
tert-butyl, i.e., 1,1-dimethylethyl, 1-1-dimethylpropyl, 
1-methyl-1-(ethyl)pentyl, 1,1-diethylpropyl, 1-ethyl-1-(propyl)butyl and 
the like. 
The term "alkyl" defines straight or branched chain monovalent hydrocarbon 
radicals having between about 1 to 6 carbon atoms including, for example, 
methyl, ethyl, propyl, isopropyl, butyl, t-butyl, pentyl, 1-methylbutyl, 
isopentyl, neopentyl, hexyl, etc. 
The terms "heterocycle" and "heterocyclic ring" as used herein refer to 
aromatic or nonaromatic heterocyclic rings which contain one or more 
heteroatoms and include but are not limited to pyridine, piperazine, 
piperidine, morpholine, azabicycloalkyl, e.g., 3-azabicyclo[3,2,2]nonane, 
azatricycloalkyl, 1,2,3,4-tetrahydroisoquinoline, 
5,6,11,12-tetrahydrodibenz[b,f]azocine, iminostilbene, and the like which 
may optionally be substituted with one or more substituents selected from 
alkyl, phenyl, substituted phenyl, phenylalkyl, heterocycle, cycloalkyl, 
halogen, hydroxy and lower alkoxy. 
The terms "substituted phenyl" and "substituted phenylalkyl" as used herein 
refer to phenyl or phenylalkyl moieties in which the phenyl ring is 
substituted by one or more substituents selected from alkyl, hydroxy, 
alkoxy, halogen, alkylamino, dialkylamino, phenyl and alkyl carbonyl. 
The term "cycloalkyl" refers to cycloalkyl rings of 3 to 10 carbon atoms 
and includes but is not limited to cyclopentyl, cyclohexyl, adamantane, 
norbornane and the like which may optionally be substituted by 1 or more 
substituents selected from alkyl, hydroxy, alkoxy, and halogen. 
The term "halogen" refers to chlorine, bromine, fluorine, and iodine. 
The expression "pharmaceutically acceptable salts" is intended to include 
those salts capable of being formed with the compounds of the present 
invention, e.g., when R represents OH, NR.sup.5 R.sup.6 or alkyl carboxyl, 
without materially altering the chemical structure or pharmacological 
properties thereof. Such salt include inorganic and organic cations or 
acid addition salts, such as sodium, potassium, calcium, ammonium, 
alkylammonium, quaternary ammonium, triethanolamine, lysine, hydrochloric, 
hydrobromide, etc. well known to those skilled in the art. The foregoing 
salts are prepared in the conventional manner by neutralization of the 
compounds of formula I with the desired base or acid. 
The compounds of the present invention can be administered to a patient in 
such oral dosage forms as tablets, capsules, pills, powders, granules, 
elixirs, or syrups as well as aerosols for inhalation. Likewise, 
administration may be effected intravascularly, subcutaneously, or 
intramuscularly using dosage forms known to those of ordinary skill in the 
pharmaceutical arts. In general, the preferred form of administration is 
oral. An effective but non-toxic amount of the compound is employed in 
treatment. The dosage regimen utilizing the present compounds is selected 
in accordance with a variety of factors including the type, age, weight, 
sex, and medical condition of the patient; the severity of the condition 
to be ameliorated; and the route of administration. A physician of 
ordinary skill can readily determine and prescribe the effective amount of 
the drug required to prevent, treat or arrest the progress of the 
condition. Dosages of the compounds of the present invention, will range 
generally between 0.1 mg/kg/day to about 100 mg/kg/day and preferably 
between about 0.5 mg/kg/day to about 50 mg/kg/day when administered to 
patients suffering from inflammation or allergic or hypersensitivity 
reactions. The compounds may also be administered transdermally or 
topically to treat proliferative skin conditions such as psoriasis. The 
daily dosage may be administered in a single dose or in equal divided 
doses three or four times daily. 
In the pharmaceutical compositions and methods of the present invention, at 
least one of the active compounds of the invention or a pharmaceutically 
acceptable salt thereof will typically be administered in admixture with 
suitable pharmaceutical diluents, excipients, or carriers (collectively 
referred to herein as "carrier" materials) suitably selected with respect 
to the intended form of administration, that is, oral tablets, capsules, 
elixirs, syrups, and the like, and consistent with conventional 
pharmaceutical practices. For instance, for oral administration in the 
form of tablets or capsules, the active drug component may be combined 
with any oral non-toxic pharmaceutically acceptable inert carrier such as 
lactose, starch, sucrose, cellulose, magnesium stearate, dicalcium 
phosphate, calcium sulfate, mannitol and the like; for oral administration 
in liquid form, the active drug component may be combined with any oral 
non-toxic pharmaceutically acceptable inert carrier such as ethanol and 
the like. Moreover, when desired or necessary, suitable binders, 
lubricants, disintegrating agents and coloring agents can also be 
incorporated in the mixture. Suitable binders include starch, gelatin, 
natural sugars, corn sweeteners, natural and synthetic gums such as 
acacia, sodium alginate, carboxymethylcellulose, polyethylene glycol, and 
waxes. Lubricants for use in these dosage forms include boric acid, sodium 
benzoate, sodium acetate, sodium chloride, and the like. Disintegrators 
include, without limitation, starch, methylcellulose, agar, bentonite, 
guar gum, and the like. 
The compounds of the invention are prepared from readily available starting 
materials by any of the following alternate processes in a conventional 
manner. The following reaction schemes describe methods which can be 
employed for preparing the compounds of formula I, including starting 
materials, intermediates, and reaction conditions. 
As shown in part (1) of Scheme A, a mercaptan (V) can be reacted with an 
oxabicyclo compound (VI) to give the intermediate (III) which may then be 
reacted with a monohaloacid in a base or alternatively with a thiol acid 
in an acid to obtain a product acid of type (VII). Epoxides of type (VI) 
are readily prepared by oxidation of a double bond with peroxides such as 
m-chloroperbenzoic acid, peracetic acid, per trifluoroacetic acids, 
hydrogen peroxide, t-butyl hydroperoxide and the like. Base induced 
cyclization of halohydrins, obtained by treatment of double bonds with 
mineral acids, also produces epoxides. In addition, epoxides can be used 
as starting materials for the preparation of halohydrins which can also be 
used to produce compounds of type III, following reaction with, for 
example, a mercaptan. Most bases can be used for the preparation of III, 
for example, hydroxides, tert-amines, heterocyclic amines, 
dimethylaminopyridines, hydrides, lithium alkyls, lithium amides and the 
like, since the thiolate anion is an exceptional nucleophile. 
Non-nucleophilic bases are preferred for the conversion of III into VII in 
the presence of an electrophilic reagent such as a substituted halo alkyl 
group. 
Compound III may also be converted into VII via conversion into a halo 
compound (Scheme C), an activated ester (Scheme C) or acid catalysis 
(Scheme A). In the first case, treatment of the hydroxy compound with 
hydrochloric, hydrobromic, hydriodic or hydrofluoric acid, preferably at 
reflux temperatures, converts it into the corresponding halo compound. 
Displacement of the halogen (SN.sub.2) with a mercaptan under basic 
conditions (as shown in Scheme B) provides compound XIII. 
The alcohols III or XI may be converted into activated esters such as those 
of toluene sulfonic acid (tosylates), methane sulfonic acid (mesylates), 
trifluoromethane sulfonic acid Triflates) and trifluoroacetic acid. 
Displacement of the activated ester (SN.sub.2) with a mercaptan under 
basic conditions (see above) provides compound VII. Both this method and 
that outlined above utilizing a halo intermediate have the advantage that 
the stereochemistry at the carbon bearing the functional group may be 
inverted thus allowing control (selection) of the stereochemistry (cis or 
trans) of the product. 
Treatment of alcohols III or XI with a mercaptan in the presence of an acid 
(Scheme A, B, C) should provide the corresponding sulfide, e.g., VII or 
XIII. Mineral acids, organic acids and Lewis acids are suitable for this 
reaction. Non-nucleophilic acids are preferred and include, for example, 
trifluoroacetic acid, toluene sulfonic acid, perfluorobutyric acid, 
triflic acid, phosphoric acid, sulfuric acid, boron trifloride, aluminum 
chloride and the like. 
Conversion of a carboxylic acid such as VII, XI or XII into an ester or an 
amide is accomplished by standard means. The carboxylic acid may be 
treated with the appropriate alcohol with or without added solvent in the 
presence of an acid (see above) to provide the product ester. A salt of 
the carboxylic acid may be prepared by treatment with a base (see above) 
and the salt then treated with an electrophilic group with displacement 
of, for example, a halide, tosylate and the like. An alternative method of 
preparation is conversion of the carboxylic acid into an activate carbonyl 
function such as an acid halide, mixed anhydride or activated ester 
followed by treatment with an appropriate alcohol or amine. Acid halides 
can be made by mixing, for example, thionyl chloride, thionyl bromide, 
phosphorus trichloride, phosphorus pentabromide, oxalyl chloride and the 
like with the acid. Mixed anhydrides with, e.g., isobutyl chloroformate, 
are prepared in the standard manner with the acid being treated with 
isobutyl chloroformate in the presence of a base or from a preformed 
carboxylate salt. The same type of salt, either prepared in the reaction 
or preformed, can be treated with for example, N-chlorosuccinimide, to 
form the succinimide activated ester. Treatment of either of these 
intermediates with the appropriate amine, alcohol, mercaptan or 
electrophile will provide the compounds of this invention. 
The conversion of the alcohols/mercaptans III, XI, XVI, XVII and the like 
into compounds of, for example, X, is accomplished in the same manner as 
the synthesis of the other esters outlined above. In this case, the 
appropriate alcohol/mercaptan is represented as outlined above and the 
acylating agent is an acid halide or anhydride. 
Scheme C illustrates yet another method for the preparation of the 
intermediates and compounds of this invention. An alpha-halo ketone, 
substituted or unsubstituted, is treated with a oxygen or sulfur 
nucleophile generated as described above. The valuable intermediate, XV, 
is reduced directly with, for example, a hydride reducing agent such as 
sodium borohydride, lithium aluminum hydride, sodium cyano borohydride and 
the like or borane, to provide the alcohols III and XI. The use of these 
intermediates for the preparation of compounds VII, VIII, XIV and X is 
discussed above. Conversion of ketone XV into a thioketone is readily 
accomplished using reagents such as phosphorus pentasulfide or 
2,4-bis(4-methoxyphenyl)-1,3-dithia-2,4-diphosphetane-2,4-disulfide 
(Lawesson's Reagent). Reduction of the thioketone as discussed above for 
the ketones provides the mercaptan analogs of III and XI and it is used 
similarly. 
##STR12## 
BIOLOGICAL EVALUATIONS 
The compounds of the invention are evaluated with respect to superoxide 
modulating activity according to the following assay procedure: 
Human neutrophil superoxide generation: Superoxide generation by 
formyl-methionyl-leucyl-phenylalanine (FMLP)-stimulated neutrophils was 
quantitated by the reduction of cytochrome C (Badwey, J. A., Curnutte, J. 
T. and Karnovsky, M. L., cis-Polyunsaturated fatty acids induce high 
levels of superoxide production by human neutrophils. J. Biol. Chem. 256: 
12640-12643, 1981.) To 5 million neutrophils in 2.85 ml of Krebs-Ringer 
phosphate buffer, pH 7.2, 50 ul of inhibitor (in 10% DMSO/buffer), and 50 
ul ferricytochrome C (5 mM, stock) were added and preincubated for 3 
minutes at 37.degree. C. Absorption measurements at 550 nm were recorded 
at start of preincubation. Fifty ul FMLP (6 uM, stock) was added to 
initiate reaction. A plateau was reached within 3 minutes and this reading 
minus the initial reading (before addition of FMLP) was used to calculate 
nanomoles of superoxide generated based on a molar extinction coefficient 
of 2.1.times.10.sup.4 cm.sup.-1 mole.sup.-1. 
Isolation of human neutrophils: Human neutrophils were isolated from 
freshly drawn blood of healthy donors. Two ml of 5% dextran (MW 
200,000-300,000) in saline was added to 10 ml aliquots of blood, mixed and 
placed upright for 45 min. at 37.degree. C. Approx. 8-10 ml of the 
plasma-white cell suspension from the dextran sedimentation was layered on 
3 ml of Ficol-paque in a 15 ml tube and centrifuged at 400 g for 30 min. 
The supernate, containing plasma and platelets, was discarded by 
aspiration, and the pellet, containing predominantly neutrophils, was 
resuspended in 1 ml saline. The suspension was transferred to a clean 
tube, and pooled with other aliquots of blood treated similarly. The 
pooled suspension was centrifuged at 350 g for 5 min. and supernate 
discarded. The pellet was resuspended in 5 ml of 0.05% NaCl with a plastic 
Pasteur pipette for 25 seconds to lyse contaminating red cells, then 5 ml 
of 1.75% NaCl added to regain isotonicity. The red cell lysing procedure 
was repeated, the cells suspended in appropriate buffer (depending on 
assay) and counted. 
The compounds of the present invention evaluated with respect to 
cyclooxygenase inhibition according to the following assay procedure. 
Inhibition of Sheep Seminal Vesicle Microsome Cyclooxygenase 
The assay was based on oxygen consumption during conversion of arachidonic 
acid to prostaglandin G.sub.2 catalyzed by cyclooxygenase Biochem. 
11:3276-3285 (1972). Lyophilized ovine microsome (approx. 1 mg) suspended 
in 2.9 ml Tris-HCl buffer, pH 8.2, containing 0.7 mM phenol were used as 
source of arachidonate cyclooxygenase. The inhibitor, 50 .mu.l in DMSO, 
was added and the mixture preincubated for 5 minutes at 37.degree. C. 
Fifty .mu.l of arachidonic acid (final conc. 50 .mu.M) was added to start 
the reaction. The slopes of the initial rates of oxygen uptake, in the 
presence and absence of inhibitor, were compared to determine reaction 
inhibition. Percent inhibition was computed using the following formula: 
##EQU1## 
The compounds of the invention are evaluated with respect to 5-lipoxygenase 
inhibition according to the following assay procedure. 
Inhibition of 5-lipoxygenase, in vitro: anti-inflammatory, anti-allergy 
activities. 
The 100,000 x g supernatant fraction of Rat Basophilic Leukemia Cell 
Homogenate (RBL-1) serves as a 5-lipoxygenase enzyme source. The enzyme is 
incubated with [1-.sup.14 C)-arachidonic acid and Ca++ in the presence and 
absence of test compound. The product of 5-lipoxygenase, 
5-hydroxyeicosatetraenoic acid (5-HETE), is separated by thin-layer 
chromatography an measured by radioactivity. A compound inhibiting 5-HETE 
synthesis by 30% or more is considered active at that concentration. 
Initial screening doses are 1.times.10.sup.-4 M. When the compound 
inhibits more than 50% of 5-HETE synthesis at 10.sup.-4 M, that compound 
is tested at multiple dose levels to determine the IC.sub.50 value 
(inhibitory concentration to inhibit 50%). 
For comparison the compound of Formula XX, a known 5-lipoxygenase inhibitor 
described in U.S. Pat. No. 4,755,524 was used. 
##STR13## 
The results with respect to certain compounds of the present invention are 
set forth in Table I below. 
TABLE 1 
______________________________________ 
Compound 
5-Lipoxygenase 
FMLP Induced Cyclooxygenase 
Example Inhibition Superoxide Inhibition 
Number IC.sub.50 (.mu.M) 
Generation IC.sub.50 (.mu.M) 
______________________________________ 
4 -- Stimulation -- 
1 .mu.M, 
67% &gt; control 
10 .mu.M, 
133% &gt; control 
100 .mu.M, 
167% &gt; control 
5 100 Inhibition -- 
IC.sub.50 = 1.1 .mu.M 
7 Stimulated 45% 
Inhibition Inactive at 
at 10.sup.-4 M 
IC.sub.50 = 50 .mu.M 
100 .mu.M 
Stimulated 27% 
at 10.sup.-5 M 
11 Stimulated Inhibition 2.9 
2.5% at 10.sup.-4 M 
IC.sub.50 = 4.8 .mu.M 
Stimulated 
3.2% 10.sup.-5 M 
15 Inhibited 21.3% 
Inhibition 64.0 
at 10.sup.-4 M 
IC.sub.50 = 2.8 .mu.M 
Inhibited 14.9% 
at 10.sup.-5 M 
16 100 Stimulation -- 
10 .mu.M, 
33% &gt; control 
25 .mu. M, 
83% &gt; control 
50 .mu.M 
100% &gt; control 
17 &gt;100 Inhibition -- 
IC.sub.50 = 3.4 .mu.M 
Formula 4.9 Inhibition -- 
XX IC.sub.50 = 11 .mu.M 
______________________________________ 
The compound of Formula XX inhibited both superoxide generation and 
5-lipoxygenase whereas the compound of Example 16 inhibited 5-lipoxygenase 
and stimulated superoxide generation. This data indicates that superoxide 
generation is not dependent on 5-lipoxygenase and that the ability of a 
compound to inhibit 5-lipoxygenase is not related to its ability to 
simulate superoxide generation. 
The compound of Example 7, which has no substituents on the phenyl ring 
(i.e., R.sup.1 and R.sup.2 =H) did not inhibit either 5-lipoxygenase or 
cyclooxygenase but did inhibit superoxide generation. 
Complement C5a induced superoxide generation may also be stimulated or 
inhibited by compounds of the present invention. 
While the invention has been described and illustrated with reference to 
certain preferred embodiments thereof, those skilled in the art will 
appreciate that various changes, modifications, and substitutions can be 
made therein without departing from the spirit of the invention. For 
example, effective dosages other than the preferred ranges set forth 
hereinabove may be applicable as a consequence of variations in the 
responsiveness of the mammal treated, severity of condition treated, 
dosage related adverse effects, if any, observed and analogous 
considerations. Likewise, the specific pharmacological responses observed 
may vary depending upon the particular active compounds selected or 
whether different active compounds are used in combination or in the 
presence of suitable pharmaceutical carriers, as well as the type of 
formulation and mode of administration employed, and such expected 
variations or differences in results are contemplated in accordance with 
the objects and practices of the present invention. It is intended, 
therefore, that the invention be limited only by the scope of the claims 
which follow. 
The following non-limiting examples further illustrate details for the 
preparation of the compounds of the present invention. Those skilled in 
the art will readily understand and appreciate that known variations of 
the conditions and procedures in the following preparative methods can be 
utilized. All temperatures are degrees Celsius unless otherwise noted. 
Melting points were determined on a Thomas-Hoover melting point apparatus 
and are uncorrected. 
EXAMPLE 1 
##STR14## 
Potassium hexylmethyl disilane (15% by weight in toluene, 260 ml, 0.169 
moles) was added by syringe to a solution of 3,5-di-tert-butylphenol (34.8 
g, 0.169 moles) in tetrahydrofuran (500 ml). After 30 minutes, a solution 
of dimethylthiocarbamoyl chloride (24.7 g, 0.20 moles) in tetrahydrofuran 
(50 ml) was added over 10 minutes. The reaction mixture was stirred at 
room temperature for 30 minutes then at 50.degree. C. for 1.5 hours. After 
cooling to room temperature, the reaction mixture was poured into cold 
(0.degree. C.) water (100 ml) containing potassium hydroxide (30 g). The 
mixture was extracted twice with 1000 ml of ethyl ether. The combined 
ethyl ether extracts were dried over sodium sulfate, filtered and 
concentrated with a rotary evaporator to give the crude product as a 
yellow oil. The title product was purified by chromatography on silica gel 
and used directly in the next Example 2. 
EXAMPLE 2 
##STR15## 
The compound of Example 1 (42 g, 0.143 moles) was heated to 300.degree. C. 
in a round bottom flask with a heating mantle for 2 hours. After cooling 
to room temperature the material was dissolved in ethylene glycol (100 
ml). A solution of potassium hydroxide (12.0 g, 0.214 moles) in water (20 
ml) was added and the reaction mixture was heated to 123.degree. C. for 
3.5 hrs. After stirring at room temperature for 20 hours, the reaction 
mixture was cooled to 0.degree. C. with an ice bath, and 10% hydrochloric 
acid was added slowly to adjust the pH to 2.0. The reaction mixture was 
extracted twice with 100 ml of ethyl acetate. The combined ethyl acetate 
extracts were washed with brine (100 ml), dried over sodium sulfate, 
filtered and concentrated to an oil. The title product was purified by 
silica gel chromatography and recrystallized from pentane, m.p. ca. 
58.degree. C. The structure assignment was supported by NMR, infrared 
spectroscopy and elemental analysis. 
Analysis calculated for: C.sub.14 H.sub.22 S (m.w.=22.4): 
Theory: C, 75.61; H, 9.71; S, 14.42. Found: C, 75.55; H, 10.07; S, 14.34. 
EXAMPLE 3 
##STR16## 
3,5-Bis(1,1-dimethylethyl)benzenethiol (Example 2) (5.0 g, 0.0225 moles) 
was added to freshly prepared sodium ethoxide (0.0230 moles) in absolute 
ethyl alcohol (50 ml). After stirring for 1 hour, cyclohexene oxide (2.2 
g; 0.0225 moles) was added by syringe over 5 minutes, and the reaction 
mixture was stirred for 60 hours at room temperature. Water (100 ml) was 
added, and the reaction mixture was extracted twice with 75 ml of ethyl 
acetate. The combined ethyl acetate extracts were washed with brine (50 
ml) dried over sodium sulfate, filtered and concentrated to give the 
product as a yellow solid which was recrystallized from cold pentane. The 
structure assignment was supported by NMR spectroscopy. The title compound 
was used in Example 4. 
EXAMPLE 4 
##STR17## 
Methyl thioglycolate (2.65 g, 0.025 moles) was added by syringe to a 
solution of trans-2-[[3,5-bis(1,1-dimethylethyl)phenyl]thio]cyclohexanol 
(5.0 g., 0.025 moles) in methylene chloride (10 ml). After stirring the 
reaction mixture for 15 minutes, trifluoroacetic acid (10 ml) was added by 
syringe, and the reaction mixture was stirred for 20 hours at room 
temperature. The reaction mixture was poured into cold water (100 ml). 
After stirring for 30 minutes, the mixture was extracted with ethyl 
acetate. The aqueous layer was washed with ethyl acetate (50 ml). The 
combined ethyl acetate extracts were washed twice with 75 ml of water, 
dried over sodium sulfate, filtered and concentrated to give the crude 
product as an oil. The title compound was purified by silica gel 
chromatography and dried in a vacuum oven at 60.degree. C. for 3 hours. 
The structure assignment was supported by NMR, infrared spectroscopy, and 
elemental analysis. 
Analysis calculated for: C.sub.23 H.sub.36 O.sub.2 S.sub.2 (m.w.=408.66): 
Theory: C, 67.60; H, 8.88; S, 15.69. Found: C, 67.62; H, 9.13; S, 15.58. 
EXAMPLE 5 
##STR18## 
Water was added to a solution of the compound of Example 4 (9.2 g, 0.0255 
moles) in methyl alcohol (100 ml) until the solution became cloudy. 
Lithium hydroxide hydrate (1.75 g, 0.0675 moles) was added, and the 
reaction mixture was stirred at room temperature. Periodically, water was 
added to make the solution cloudy. After 6 hours, the solution was made 
acidic with 10% hydrochloric acid and extracted with ethyl acetate. The 
ethyl acetate extract was dried over sodium sulfate, filtered, and 
concentrated to give the crude product as an oil. The product was purified 
by silica gel chromatography. The structure assignment was supported by 
NMR and elemental analysis. 
Analysis calculated for: C.sub.22 H.sub.34 S.sub.2 O.sub.2 (m.w.=394.63): 
Theory: C, 66.96; H, 8.68; S, 16.25. Found: C, 66.92; H, 8.80; S, 16.00. 
EXAMPLE 6 
##STR19## 
Thiophenol (2.14 g, 0.0194 mole) was added to freshly prepared sodium 
ethoxide (sodium, 0.45 g) in ethanol (30 ml). After several minutes, 
cyclohexene oxide was added, and the reaction mixture was stirred at room 
temperature for 24 hours. The reaction mixture was concentrated with a 
gentle flow of nitrogen gas. The residue was dissolved in diethyl ether 
(75 ml) and washed with 1N hydrochloric acid (19 ml). The diethyl ether 
was washed three times with 50 ml of 5% sodium carbonate, once with 0.5N 
hydrochloric acid (50 ml) and once with brine (25 ml), dried over 
anhydrous magnesium sulphate, filtered and concentrated with a rotary 
evaporator to give the product as an oil. The structure was supported by 
NMR and infrared spectroscopy. 
EXAMPLE 7 
##STR20## 
Mercaptoacetic acid (0.44 g, 0.0048 mole) was added to a cold solution of 
the compound of Example 6 (1.0 g 0.0048 mole) in methylene chloride (5 ml) 
containing trifluoracetic acid (3.5 ml). The reaction mixture was stirred 
at room temperature for 20 hours. The reaction mixture was concentrated to 
an oil with a rotary evaporator. The residue was dissolved in diethyl 
ether (50 ml), washed three times with 20 ml of 5% sodium bicarbonate, 
followed by 1N hydrochloric acid (10 ml) and water (20 ml), dried over 
anhydrous magnesium sulfate, filtered and concentrated to a colorless oil 
with a rotary evaporator. The product was purified by silica gel 
chromatography. The structure was supported by NMR, infrared spectroscopy 
and elemental analysis. 
Analysis calculated for: C.sub.14 H.sub.18 S.sub.2 O.sub.7 (m.w.=282.44): 
Theory: C, 59.54; H, 6.42; S, 22.70. Found: C, 59.36; H, 6.57; S, 22.43. 
EXAMPLE 8 
##STR21## 
meta-t-Butyl phenol (30.0 g, 0.20 mole) was added to water (140 ml) 
containing potassium hydroxide (11.2 g, 0.20 mole) and stored at 0.degree. 
C. for 20 hours. Dimethyl thiocarbamoyl chloride (32.8 g, 0.265 mole) was 
added as a solution in tetrahydrofuran (60 ml) to the cold solution with 
stirring. The ice bath was removed, and the turbid solution was stirred 
for 15 minutes. To this mixture was added 10% potassium hydroxide (75 ml). 
The reaction mixture was extracted three times with 125 ml of benzene. The 
combined benzene extracts were washed with brine (75 ml) and concentrated 
in a rotary evaporator to give the crude product as an oil. The product 
was purified by silica gel chromatography. The structure was supported by 
NMR. 
EXAMPLE 9 
##STR22## 
Starting with O-[3-(1,1-dimethylethyl)phenyl]dimethylcarbamothioate and 
following the procedure described in Example 2, the title compound was 
obtained. 
EXAMPLE 10 
##STR23## 
Using the method of Example 6 and substituting 
3-(1,1-dimethylethyl)benzenethiol for thiophenol, the title compound was 
obtained. 
EXAMPLE 11 
##STR24## 
Trifluoracetic acid (10 ml) was added to a solution of the compound of 
Example 10 (3.6 g, 0.0136 mole) in methylene chloride (5 ml) with 
stirring. After several minutes, methyl thioglycolate (1.59 g, 0.015 mole) 
was added, and the reaction mixture was stirred for 30 minutes. The 
reaction mixture was poured into methanol (50 ml) containing lithium 
hydroxide hydrate (12.6 g, 0.30 mole). Water (125 ml) was slowly added to 
the mixture and then the mixture was extracted with diethyl ether (100 
ml). The aqueous phase was acidified with concentrated hydrochloric acid 
and extracted twice with 100 ml of diethyl ether. The combined diethyl 
ether extracts were washed with water (30 ml), 5% sodium bicarbonate (50 
ml) and brine (50 ml), dried over anhydrous magnesium sulfate, filtered 
and concentrated with a rotary evaporator to give the crude product as an 
oil. The product was purified by silica gel chromatography. The structure 
was supported by NMR, infrared spectroscopy, and elemental analysis. 
Analysis calculated for: C.sub.18 H.sub.26 S.sub.2 O.sub.2 (m.w.=338.52): 
Theory: C, 63.87; H, 7.74; S, 18.94. Found: C, 64.00; H, 8.02; S, 19.10. 
EXAMPLE 12 
##STR25## 
Using the method of Example 8 and substituting meta-phenyl phenol for 
meta-t-butyl phenol the title compound was prepared. The structure was 
supported by NMR. 
EXAMPLE 13 
##STR26## 
Starting with [1,1'-biphenyl]-3-yl dimethylcarbamoate and using the 
procedure described in Example 2, the title compound was obtained. 
EXAMPLE 14 
##STR27## 
Starting with 3-[1,1'-biphenyl]thiol and following the procedure described 
in Example 3 gave the title compound. 
EXAMPLE 15 
##STR28## 
Starting with trans-2-[([1,1'-biphenyl]-3-yl)thio]cyclohexanol and using 
the method described in Example 11 gave the title compound. The structure 
was supported by NMR, infrared spectroscopy and elemental analysis. 
Analysis calculated for: C.sub.20 H.sub.22 S.sub.2 O.sub.2 (m.w.=358.51): 
Theory: C, 67.00; H, 6.18; S, 17.89. Found: C, 66.94; H, 6.30; S, 18.07. 
EXAMPLE 16 
##STR29## 
Oxalyl chloride (0.19 g, 0.0015 moles) was added by syringe to a cold 
(10.degree. C.) solution of 
trans-[[2-[[3,5-bis(1,1-dimethylethyl)phenyl]thio]cyclohexyl]thio]acetic 
acid (Example 5) (0.55 g, 0.0014 moles) in benzene (50 ml). The cold bath 
was removed and the reaction mixture was stirred at room temperature for 
20 hours. The reaction mixture was concentrated to an oil using a rotary 
evaporator. The oil was dissolved in toluene (50 ml) and concentrated to 
an oil. The process was repeated using tetrahydrofuran (25 ml) instead of 
toluene. The residue was dissolved in tetrahydrofuran (50 ml). To this 
solution was added 2-(2-methylaminoethyl)pyridine (0.19 g, 0.0014 moles) 
and triethylamine (0.22 g) and the reaction mixture was stirred at room 
temperature for 48 hours. The white solid precipitate was removed by 
filtration and washed with ethyl acetate (25 ml). The filtrate was 
concentrated to give the crude product as an oil. The produce was purified 
by silica gel chromatography and dried in vacuo at 100.degree. C. for 1 
hour to give the title compound. The structure assignment was supported by 
NMR, infrared spectroscopy and elemental analysis. 
Analysis calculated for: C.sub.30 H.sub.44 N.sub.2 O.sub.2 S.sub.2 
(m.w.=512.83): 
Theory: C, 70.26; H, 8.65; S, 5.45. Found: C, 69.96; H, 8.76; S, 5.43. 
EXAMPLE 17 
##STR30## 
Sodium hydride (0.33 g, 0.0138 mole) was added to a solution of 
trans-2-[[3,5-bis(1,1-dimethylethyl)phenyl]thio]cyclohexanol (3.4 g, 
0.0106 mole) in tetrahydrofuran (50 ml) at 0.degree. C. After stirring the 
reaction mixture for 1.5 hr, the tetrahydrofuran was removed by rotary 
evaporation. Dimethyl sulfoxide (75 ml) was added followed by chloroacetic 
acid sodium salt (1.48 g, 0.0127 mole) and the reaction mixture was 
stirred at room temperature for 10 days. Water (100 ml) was added dropwise 
to the mixture followed by 10% hydrochloric acid (10 ml). The product was 
extracted twice with 200 ml of ethyl acetate. The combined ethyl acetate 
extracts were washed twice with 200 ml of water, dried over anhydrous 
sodium sulfate, filtered, and concentrated. The product was purified by 
chromatography on silica gel. The structure was supported by NMR and 
elemental analysis (378.6+1/4 mole H.sub.2 O). 
Analysis calculated for: C.sub.22 H.sub.34 O.sub.3 S+1/4 mole H.sub.2 O: 
Theory: C, 68.98; H, 9.08; S, 8.37. Found: C, 69.12; H, 9.21; S, 8.27. 
EXAMPLE 18 
##STR31## 
2,5-Dihydrofuran (DHF) (13.2 g, 0.188 mole) was added by syringe to a 
solution containing 3-chloroperoxybenzoic acid (29.1 g, 0.198, mole) and 
trifuloroacetic acid (0.5 ml) in methylene chloride (500 ml). After 
stirring at room temperature for 20 hours, the white solid was removed by 
filtration. The filtrate was washed with a solution of sodium carbonate 
(100 ml, saturated). The organic phase was stirred with solid sodium 
carbonate and sodium thiosulfate for 20 minutes and filtered. The product 
was purified by low pressure distillation (41.degree. C./5 mmHg). The 
structure was supported by NMR. 
EXAMPLE 19 
##STR32## 
2,6-bis(1,1-Dimethylethyl)-4-benzenethiol (0.0078 mole) and the compound of 
Example 18 (0.0074 mole) are added to a degassed (Argon) solution of 50% 
sodium hydroxide (5 ml) and isopropyl alcohol (50 ml). The reaction is 
heated to reflux for 24 hours. The reaction is cooled to room temperature 
and poured into water (125 ml). The solution is made acidic with 1N 
hydrochloric acid and extracted 3 times with 100 ml of diethyl ether. The 
combined diethyl ether extracts are dried over anhydrous magnesium 
sulfate, filtered and concentrated with a rotary evaporator. The product 
is purified by silica gel chromatography. 
EXAMPLE 20 
##STR33## 
The compound of Example 19 (0.0062 mole) is added to acetic anhydride (20 
ml). Triethylamine (0.0062 mole) is added, and the reaction mixture is 
stirred for 3 hours. Additional triethylamine (0.3 ml) is added, and the 
reaction mixture is stirred for 2 hours. The reaction mixture is 
concentrated to an oil with a gentle flow of nitrogen gas. The residue is 
dissolved in diethyl ether (75 ml), washed twice with 50 ml of 0.25N 
hydrochloric acid and once with 25 ml of brine, dried over anhydrous 
magnesium sulfate, filtered and concentrated with a gentle flow of 
nitrogen gas. The product is purified by silica gel chromatography. 
EXAMPLE 21 
##STR34## 
Succinic anhydride (0.0185 mole) and triethylamine (0.0185 mole) are added 
to a solution of tetrahydrofuran (THF) (50 ml) containing the compound of 
Example 19 (0.0092 mole). The reaction mixture is stirred for 3 days and 
then concentrated to an oil with a gentle flow of nitrogen gas. The 
residue is dissolved in diethyl ether. The solution is washed twice with 
50 ml of water and once with 20 ml of 1N hydrochloric acid, dried over 
anhydrous magnesium sulfate, filtered and concentrated to an oil with a 
rotary evaporator. The product is purified by silica gel chromatography.