NOVEL COMPOUNDS AND METHODS FOR TREATING MULTIPLE SCLEROSIS

This invention relates to novel thiomorpholinone compounds useful for treating multiple sclerosis.

Multiple Sclerosis was first described as a clinical entity in 1868. Clinically, it is a highly variable disease, which usually begins between the second and fifth decades of life. The most common signs of multiple sclerosis are sensory and visual motor dysfunction. In the chronic form the patient has periods of remission, but with each remission there is greater neurological dysfunction.

Macroscopically, multiple sclerosis involves lesions of 1 to 4 cm called plaques scattered throughout the white matter of the central nervous system. Microscopically, the disease is characterized by a break down of the nervous system's myelin sheath. There is also a loss of myelin basic protein in the area of the lesions.

The etiology and pathogenesis of multiple sclerosis remains obscure. Both chronic infectious agents and autoimmunity have been involved and, m fact, both might be important .

Meanwhile, the need continues for safer, better calibrated drugs which will either slow the process of neurodegeneration associated with multiple sclerosis or even prevent such neurodegeneration altogether. The present invention provides new thiomorpholmone compounds useful for treating multiple sclerosis. These compounds provide for safe and efficacious treatment of multiple sclerosis by slowing the process of neurodegeneration associated with such disease.

This invention provides compounds of the formula (I)

wherein:

R ¹ and R^ are each independently selected from Ci-Cβ alkyl; C2-C8 alkenyl; C2-C8 alkynyl; Ci-Cβ alkyloxy; C1-C8 alkylthio; trifluoromethyl; C1-C4 alkyl substituted with phenyl; phenyl; F; Cl; NO2; phenoxy; C1-C4 alkyl substituted with phenoxy; thiophenyl; C3 _. -C4 alkylthiophenyl; -COOR ⁷ ; -N(R ⁷ )2 or -N(R ⁷ )Sθ2R ⁷ where each R ⁷ is independently hydrogen or Ci-Cβ alkyl;

R ³ is H or C1-C4 alkyl;

R and R^ are each individually H, or when taken together form a bond;

R ⁶ is H; C1-C8 alkyl; C2-C8 alkenyl; C2-C8 alkynyl; -SO2CH3; - (CH2) _n NR ⁸ R ⁹ ; - (CH2) _n C02R ⁸ ; -(CH2) _n OR ⁸ where n is an

integer from 1 to 6, both inclusive, and R ⁸ and R ⁹ are each independently hydrogen; Ci-Cg alkyl; C2-C6 alkenyl; C2-C6 alkynyl; phenyl; C1-C4 alkyl substituted with phenyl; -(CH2)qOH; or (CH2)qS (Cι-C4alkyl) where q is an integer from 2 to 6, both inclusive; and

X is - °S*- where m is 0 or 1; or a pharmaceutically acceptable salt or optical isomer thereof .

According to a further aspect of the present invention there are provided pharmaceutical compositions comprising as active ingredient a compound of Formula I or a pharmaceutically acceptable salt thereof, m association with one or more pharmaceutically acceptable diluents, carriers and excipients thereof. The present invention also provides a method for treating multiple sclerosis m a mammal in need of such treatment, which comprises administering to said mammal a therapeutically effective amount of a compound, or pnarmaceutically acceptable salt or isomer thereof, of the formula I.

Other objects, features and advantages of the present invention will become apparent from the subsequent description and the appended claims.

As used herein, the term "C1-C8 alkyl" represents a straight or branched alkyl chain having from one to eight carbon atoms. Typical Cι_-C8 alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, t- butyl, n-pentyl, isopentyl, neopentyl, heptyl, hexyl, octyl and the like. The term "C1-C8 alkyl" includes within its definition the terms "C1-C4 alkyl" and "Cχ-C6 alkyl' .

"C1-C4 alkyl substituted with phenyl" represents a straight or branched chain alkyl group having from one to four carbon atoms attached to a phenyl ring. Typical C1-C4 alkylphenyl groups include benzyl, phenylethyl, phenyIpropyl, 1-methyl-l-phenylethyl, phenylbutyl, 2-methyl- 3-phenyIpropyl, and 1, 1-dimethyl-2-phenylethyl .

The term "C1-C4 alkylthiophenyl" represents a straight or branched chain alkyl group having from one to four carbon atoms attached to a thiophenyl moiety. Typical C1-C4 alkylthiophenyl groups include methylthiophenyl, 2- methylethylthiophenyl, ethylthiophenyl, isobutylthiophenyl and the like.

In a similar fashion, the term "C1-C4 alkyl substituted with phenoxy" represents a straight or branched chain alkyl group having from one to four carbon atoms substituted with a phenoxy moiety. Typical C1-C4 alkyloxyphenyl groups include phenoxymethyl, phenoxyethyl, phenoxypropyl and the like.

"C1-C8 alkyloxy" represents a straight or branched alkyl chain having one to eight carbon atoms, which chain is attached to the remainder of the molecule by an oxygen atom. Typical Cι_-C8 alkyloxy groups include methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert- butoxy, pentoxy, hexoxy, heptoxy, and the like. The term "C1-C8 alkoxy" includes within its definition the term "Ci- C4 alkoxy" .

"C1-C8 alkylthio" represents a straight or branched alkyl chain having one to eight carbon atoms, which chain is attached to the remainder of the molecule by a sulfur atom. Typical C1-C8 alkylthio groups include methylthio, ethylthio, propylthio, butylthio, tert- butylthio, octylthio and the like. The term "Cχ-C8

alkylthio" includes within its definition the term ' C1-C4 alkylthio" .

The term "C2-C6 alKenyl refers to straight and branched chain radicals of two to six carbon atoms, both inclusive, having one or more double bonds. As such, the term includes ethylene, propylene, l-butene, 2-butene, 2-methyl-l-propene, 1-pentene, 2-methyl-2-butene and the like.

The term "C2-C6 alkynyl" refers to straight and branched chain radicals of two to six carbon atoms, both inclusive, having one or more triple bonds. As such, the term includes acetylene, propyne, 1-butyne, 2-hexyne, 1- pentyne, 3-ethyl-l-butyne and the like.

The term "pharmaceutically acceptable salts" refers to salts of the compounds of the above formulae which are substantially non-toxic to living organisms. Typical pharmaceutically acceptable salts include those salts prepared by reaction of the compounds of the above formulae with a pharmaceutically acceptable mineral or organic acid, or a pharmaceutically acceptable alkali metal or organic base, depending on the types of substituents present on the compounds of the formulae.

Examples of pharmaceutically acceptable mineral acids which may be used to prepare pharmaceutically acceptable salts include hydrochloric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, phosphorous acid and the like. Examples of pharmaceutically acceptable organic acids which may be used to prepare pharmaceutically acceptable salts include aliphatic mono and dicarboxylic acids, oxalic acid, carbonic acid, citric acid, succinic acid, phenyl-substituted alkanoic acids, aliphatic and aromatic sulfonic acids and the like. Such pharmaceutically acceptable salts prepared from mineral or organic acids thus

include hydrochloride, hydrobromide, nitrate, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, hydroiodide, hydrofluoπde, acetate, propionate, formate, oxalate, citrate, lactate, p- toluenesulfonate, methanesulfonate, maleate, and the like. Many compounds of formulae I which contain a carboxy group may be converted to a pharmaceutically acceptable salt by reaction with a pharmaceutically acceptable alkali metal or alkaline-earth metal or organic or inorganic base.

Examples of pharmaceutically acceptable alkali or alkaline- earth metal bases include compounds of the general formula MOR13 , where M represents an alkali or alkaline earth metal atom, e.g. sodium, potassium, lithium, calcium or barium and R13 represents hydrogen or C1-C4 alkyl. Examples of pharmaceutically acceptable organic and inorganic bases which may be used to prepare pharmaceutically acceptable salts include sodium carbonate, sodium bicarbonate, ammonia, amines such as triethanolamine, triethylamine, ethylamine, and the like.

It should be recognized that the particular anion or cation forming a part of any salt of this invention is not critical, so long as the salt, as a whole, is pharmacologically acceptable and as long as the anion or cationic moiety does not contribute undesired qualities. Depending upon the definitions of R-*-, R^ and R3 , the compounds of Formula I may exist in various isomeric forms. This invention is not related to any particular isomer but includes all possible individual isomers and racemates.

Preferred Compounds of the Invention

A preferred genus of compounds includes those compounds wherein R ¹ , R ² , R3 , R ⁴ , R ⁵ and m are as set forth for formula I, and R^ is hydrogen, Cι_-C6 alkyl, C2-C6 alkenyl or -(CH2)n°R ⁸ where n is 2 and R ⁸ is hydrogen.

Of this preferred genus, those compounds in which

X is -S- where m is 0 are more preferred.

Of this more preferred genus, those compounds m which R° is hydrogen are especially preferred.

Of this especially preferred genus, those compounds m which R ¹ and R ² are each independently C1-C8 alkyl; C1-C8 alkyloxy; C1 _. -C4 alkyl substituted with phenyl; phenyl; F; Cl; O2; phenoxy; C1-C4 alkylthiophenyl; -COOR ⁷ or -N(R ⁷ )S02R ⁷ , where each R ⁷ is independently hydrogen or C1-C6 alkyl, are particularly preferred.

Of this particularly preferred genus, those compounds m which R ¹ and R ² are each independently Cχ-C8 alkyl (especially C1-C4 alkyl) , or C1-C8 alkoxy (especially C1-C6 alkoxy), are more particularly preferred.

The skilled artisan w ll readily recognize that compounds encompassed by the instant invention include

2-[[3,5-bιs(l, 1-dιmethylethyl) -4-hydroxyphenyl]methylene] -4-

(2-hydroxyethyl) -3-thiomorpholmone; 2-[[3,5-bιs(l, 1-dimethylethyl) -4-hydroxyphenyl]methylene] -4-

[2- (dimethylamino) ethyl] -3-thiomorpholmone;

2-[[3,5-bιs(l, 1-dimethylethyl) -3-hydroxyphenyl]methylene] -4-

(methylsulfonyl) -3-thiomorpholmone ^•

2- [ (4-hydroxy-3 , 5-dι-2-propenylphenyl)methylene] -4-methyl-3- thiomorpholmone;

2- [ (4-hydroxy-3 , 5-dmιtrophenyl)methylene] -4-methyl-3- thiomorpholmone;

2- [ (3 , 5-dichloro-4-hydroxyphenyl)methylene] -4-methyl-3- thiomorpholmone; 2- [ [3-ethoxy-4-hydroxy-5-

[ (phenylthio)methyl]phenyl]methylene] -4-methyl-3- thiomorpholmone;

2-[[3,5-bιs(l, 1-dimethylethyl) -4-hydroxyphenyl]methylene] -4- (2-propenyl) -3-thiomorpholmone; 2-[[3,5-bιs(l, 1-dimethylethy1-4-hydroxyphenyl]methylene] -3- thiomorpholmone

2- [ [ [ -hydroxy-5 (1-penteny1) -3-phenyl]phenyl]methylene] -4- methyl-3-thiomorpholmone; ±2 - [ [ 3 - (2-pentynyl) -4-hydroxy-5-phenoxyphenyl]methyl] -4- methyl-3-thiomorpholmone;

±2 - [ [2-trifluoromethyl-4-hydroxy-3- (2,3- dimethylpentyl)phenyl]methyl] -4-methyl-3-thiomorpholmone;

2- [ [ (3-fluoro-4-hydroxy-6-methylthio)phenyl]methylene] -4- methyl-3-thiomorpholmone; ±2- [ [ (4-hydroxy-2-methyl-6-thιophenyl)phenyl]methyl] -4- methyl-3-thiomorpholmone;

2- [ (3 , 5-diethylamino-4-hydroxyphenyl)methylene] -4-methyl-3- thiomorpholmone;

2- [ [ [3- (2-pentynyl) -4-hydroxy-5-phenoxypropyl]phenyl]methyl ^' 4-methyl-3-thiomorpholmone;

2- [ [ [4-hydroxy-2 , 6-bis (sulfonamido) ]phenyl]methylene] -4- methyl-3-thiomorpholmone;

±2 - [ [ (3-carboxy-4-hydroxy-5-dιmethylethyl)phenyl]methyl] -4- methy1-3-thiomorpholmone; 2-[ (4-hydroxy-2-methoxycarbonyl-6-ethylphenyl)methylene] -4- methy1-3-thiomorpholmone;

±2 - [ [ [4-hydroxy-3-methyl-5 [N-propyl-N- methy1sulfonylam o]phenyl]methyl] -4-methyl-3- thiomorpholmone;

2- [ [3 , 5-bιs (1, 1-dιmethylethyl) -4-hydroxyphenyl] ethylene] -4- isopentyl-3-thiomorpholmone;

2- [ [3, 5-bιs (1, 1-dιmethylethyl) - -hydroxyphenyl]methyl ] -4- (2- hexynyl) -3-thiomorpholmone;

2- [ [3 , 5-bis (1, 1-dιmethylethyl) -4-hydroxyphenyl]methylene] -4- ethynyl-3-thiomorpholmone; 2- [ [3 , 5-bιs (1, 1-dιmethylethyl) -4-hydroxyphenyl]methyl] -4- [4-

(dipnenylamino)butyl] -3-thiomorpholmone hydrochloride salt; 2-[[3,5-bιs(l, 1-dimethylethyl) - -hydroxyphenyl]methylene] -4-

[2- (N,N-bis (2-propenyl)aminoethyl-3-thiomorphol one; 2- [ [3, 5-bis (1,1-dιmethylethyl) -4-hydroxyphenyl]methyl] -4- [3- (diethynylammo)propyl] -3-thiomorpholmone oxaylic ac d salt; 2- [ [3 , 5-bιs (1, 1-dιmethylethyl) -4-hydroxyphenyl]methyl] -4- [ [3-

(2-hydroxyethyl) ammo]propyl] -3-thiomorpholmone;

2- [ [3 , 5-bis (1, 1-dimethylethyl) -4-hydroxyphenyl]methyl] -4-

[ [ [3- (N,N-dimethylamino) ethyl] ammo]propyl] -3- thiomorpholmone citric acid salt;

2- [ [3 , 5-bιε (1, 1-dιmethylethyl) -4-hydroxyphenyl]methylene] -4-

[ [3-ethylphenylammo]propyl] -3-thiomorpholmone hydrochloride salt;

±2- [ [3 , 5-bis (1, 1-dιmethylethyl) -4-hydroxyphenyl]methyl] -4- [2- (carboxyethyl) ethyl] -3-thiomorpholmone;

±2- [ [3 , 5-bis (1, 1-dιmethylethyl) -4-hydroxypheny1]methyl] -4- [2-

(ethoxycarbonyl) ethyl] -3-thiomorpholmone sodium salt;

2- [ [3 , 5-bis (1, 1-dιmethylethyl) -4-hydroxyphenyl]methylene] -4-

[3- [2- (methylthio) ethyl]am o]propyl-3-thiomorpholmone; 2- [ [ , -bis (1, 1-dιmethylethyl) -4-hydroxyphenyl]methylene] -4-

(3-ethoxypropyl) -3-thiomorpholmone;

2- [ [3 , 5-bis (1, 1-dιmethylethyl) -4-hydroxyphenyl] - ethyImethy1ene] -4-methyl-3-thiomorpholmone.

2- [ [3 , 5-bis (1, 1-dimethyle hyl) -4-hydroxyphenyl] -ethylmethyl^ 4-methyl-3-thiomorpholinone.

Synthesis Methods

Scheme I

The compounds of formula I, where R^ is hydrogen, are prepared according to the reaction scheme I outlined below.

In the above reaction scheme, an appropriately substituted phenol, dissolved in an aprotie polar solvent such as tetrahydrofuran (THF), is reacted with a molar excess of a metal oxide, such as silver oxide, to prepare the substituted quinone methide (1) . The reaction is conducted at temperatures of from about 0°C to reflux, preferably at about 25°C.

A 4-substituted 3-thiomorpholmone starting material (2) can be readily prepared by reacting 3-thiomorpholmone with an appropriately substituted halide sucn as methyl or ethyl iodide, or methylsulfonyl chloride. The substituted thiomorpholmone starting material (2) is then treated with a molar excess of an organolithium base, such as n-butyl lithium, to produce the lithium enolate. The reaction is conducted in an aprotie polar solvent, such as THF, at a temperature of from about 0°C to about 25°C preferably about 25°C.

The resultant lithium enolate solution is then reacted with the quinone methide (1) at a temperature of from about 0°C to reflux, preferably at about 25°C to produce (3) . The desired product (3) can be chromatographically purified using silica gel as the stationary phase and ethyl acetate hexane as the mobile phase.

Depending on the substituents at R*-- , R^ and R^, the compounds of the present invention may have one, two or three stereocenters. The method and compounds of the present invention encompass the diastereomers and the racemates and their individual stereoisomers. Diaεteromeric pairs may be readily separated by standard techniques such as chromatography or crystallization. The stereoisomers may be obtamed accordmg to procedures well known m that art. For example, the racemate can be resolved by treating the mixture with diisopropyl tartrate and t-butyl hydroperoxide, as described m United States Patent No. 5,216,002, herein incorporated by reference.

Scheme II

The compounds of formula I, where R ¹ -* and R^ taken together form a bond, are prepared accordmg to the following general procedure.

(3) In the above Scheme II, an appropriately substituted p- hydroxybenzaldehyde or alkyl-p-hydroxyphenyl ketone is dissolved in a halogenated hydrocarbon such as dichloromethane and then reacted at room temperature with triethylamine and an acylating agent of the formula ZCOCl where Z is the residue of a protecting group, such as pivaloyl chloride or acetyl chloride, to prepare a protected benzaldehyde or phenyl ketone starting material (1) . The lithium enolate of a 4-substituted-3- thiomorpholinone, prepared as described in Scheme I above, is then dissolved in an aprotie solvent such as THF and the solution is cooled to from about -78° to about -20°C, preferably about -78°C. The enolate solution is reacted with

13 a molar excess of the protected benzaldehyde (1) to produce the thiomorpholmone (3) .

Oxidation of the thiomorphol ones prepared by the above schemes to produce the sulfoxide can be readily accomplished by treating the thiomorpholmone with an oxidant such as metachloroperbenzoic acid (m-CPBA) at temperatures of from about -20°C to about 25°C. The reaction is preferably carried out a halogenated hydrocarbon solvent such as methylene chloride One mole of m-CPBA, per mole of (3) , is required to produce the sulfoxide, and the reaction is substantially complete within an hour.

The sulfoxide can be purified us g standard recrystallization procedures m a suitable organic solvent such as ethyl acetate/hexane. Further recrystalization may be accomplished with dichloromethane/hexane to give the desired product as the dichloromethane solvate.

Preparation of the 4-alkyl-3-thiomorpholmone starting material (2) can be achieved by an N-alkylation reaction using a suitable R^ containing halide, such as iodide or bromide, to provide the corresponding N-substituted derivatives; i.e., those compounds of formula I where R° is Cι-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, -SO2CH3 , - (CH2) _n NR ⁸ R ⁹ , -(CH2) _n C02 ⁸ where n is an integer from 1 to 6, both inclusive and R ⁸ and p are each independently hydrogen, Cχ-Cβ alkyl, C2-Cς alkenyl, C2~Cg alkynyl, phenyl, C1 _. -C4 alkylphenyl, -(CH2)q0H, - (CH2) qN(Cχ-C4alkyl) 2 or (CH2)qS(Cι- C4alkyl) where q is an integer from 2 to 6, both inclusive. For example, 4-methyl-3-thiomorpholmone can be prepared by treating a solution of 3-thiomorpholmone THF with a 60% NaH dispersion followed by methyl iodide.

It will be readily appreciated by one skilled in tne art that the substituted benzaldehyde, hydroxyphenylketone and substituted phenol starting materials are either commercially

available or can be readily prepared by known techniques from commercially available starting materials. For example, p- hydroxybenzaldehyde may be alkylated under Fπedel-Crafts conditions to yield an alkylated benzaldehyde which may in turn itself be alkylated.

Compounds where R-*- and R ² in the starting material area each independently selected from COOR ⁷ , -N(R ⁷ )2 or -N(R ⁷ ) SO2 ⁷ , where R ⁷ is hydrogen, will require the use of a protecting group, as described m the standard text "Protecting Groups m Organic Synthesis," 2nd Edition (1991), by Greene and erts. The protecting group may be readily removed by conventional techniques after the final (linkage) step. Am e protecting groups may include acetyl while t- butyl may be used to protect acid functionalities. All other reactants used to prepare the compounds in the instant mvention are commercially available.

The following examples further illustrate the preparation of the compounds of this invention. The examples are illustrative only and are not intended to limit the scope of the invention any way.

Example 1

Preparation of 2- [ [3 , 5-Bιs (1, 1-dιmethylethyl) -4- hydroxyphenyl]methyl] -4-methyl-3-thiomorpholmone.

A. 4-methyl-3-thiomorpholmone

To a stirred solution of 3-thiomorpholmone (3.38g, 28.8mmol) m tetrahydrofuran (288ml) was added 60% sodium hydride dispersion (1.27g, 31.7mmol) followed by methyl iodide (1.80ml, 28.8mmol) . After stirring for 2 hours, the reaction was quenched with water and concentrated m vacuo . The resultmg aqueous suspension was diluted with

dichloromethane and acidified with IN hydrochloric acid. Drying over sodium sulfate and evaporation of the dichloromethane layer gave a solid which was chromatographed on silica gel. Elution with 6L of a 10-50% acetone in hexane gradient yielded 4-methyl-3-thiomorpholmone (3.02g, 80%) . -•-H NMR (CDCI3) 3 3.65 (t, J=6Hz, 2H) , 3.35 (s, 2H) , 3.05 (s, 3H) , 2.9 (t, J=6Hz, 2H) .

B. 2-[[3,5-Bιs(l, 1-dιmethylethyl) -4-hydroxyphenyl]methyl] - 4-methyl-3-thiomorpholmone

4-methyl-3-thiomorpholmone (1.88g, 14.3mmol) dissolved in tetrahydrofuran (72ml) was cooled to 0'C and treated with 1.37M n-butyl lithium m hexane (20.9ml, 28.7mmol) . After five mmutes, the cold bath was removed. Ten mmutes later a solution of 2, 6-dι-t-butyl-qumone methide, prepared by adding silver oxide (33.2g, 143mmol) to 2, 6-dι-t-butyl-4- methyl-phenol (BHT) (3.16g, 14.3mmol) m tetrahydrofuran (72ml) stirring for 30 mmutes and filtering, was added dropwise over 20mm. The reaction was stirred for three hours, quenched with IN hydrochloric acid, and evaporated to an aqueous suspension. The crude product was extracted into ethyl acetate, dried over sodium sulfate and concentrated m vacuo . Chromatography on silica gel eluting with 8L of a 0- 50% ethyl acetate hexane gradient yielded desired product (2.0g, 40%) :

^ NMR (CDCI3) 3 7.1 (ε, 2H) , 5.15 (ε, IH) , 3.7-3.4 (m, 4H) , 3.1 (s, 3 H) , 2.85 (dd, ,7=15, 9Hz, IH) , 2.75 (t, J=6Hz, 2H) , 1.45 (s, 18H); FD MS 349 (M ⁺ ) ; Elemental Analysis for C20 ^H 3lNO2S

Theory: C, 68.72; H, 8.94; N, 4.01. Found: C, 68.95; H, 8.92; N, 4.23.

Example 2

Preparation of (-)2- [ [3 , 5-bιs (1, 1-dιmethylethyl) -4- hydroxyphenyl]methyl] -4-methy1-3-thiomorphol one.

To a stirred εuεpenεion of 4A molecular sieves (1.05g) in methylene chloride (25ml) was added titanium tetraisopropoxide (0.45ml, 1.5mmol) , (+)diisopropyl tartrate (0.63ml, 3.0mmol), and deionized water (27ml, 1.5mmol) , respectively. The suεpension was allowed to stir at room temperature for 20 mmutes before addition of 2- [[3,5- bis (1, 1-dimethylethyl) -4-hydroxyphenyl]methyl] -4-methyl-3- thiomorphol one (0.87g, 2.5mmol) . After dissolution of the sulfide, the reaction was cooled to -20°C and 2.57M t-butyl hydroperoxide solution in isooctane (0.58ml, 1.5mmol) was added. The reaction was stirred at -20°C for 6 hours, at which time the molecular sieves were removed by filtration. The filtrate was quenched by pouring mto a εtirred 50ml solution prepared from citric acid monohydrate (3.3g) , ferrous sulfate heptahydrate (9.9g) , and deionized water.

Stirring was continued for 30 mmutes, then the layers were left to separate. The aqueous layer was extracted with an equal volume of methylene chloride. The original methylene chloride layer and the methylene chloride extract were combined and dried over sodium sulfate. Evaporation of the solvent followed by NMR analysis of a deuterated chloroform (CDCl) solution of the reεidue showed a 31/69 ratio of starting material to sulfoxide products (67/33 mixture of sulfoxide diastereomers) . The evaporation residue was chromatographed on silica gel. Elution with 6L of a 10-50% ethyl acetate m hexane gradient yielded optically enriched starting material (0.26g, 29% recovery) as a white foam: ee 34% (HPLC) ,*

Elemental Analysis for C20 ^H 31 ^NO 2 ^s Theory: C, 68.72; H, 8.94; N, 4.01. Found: C, 68.95; H, 8.92; N, 4.23.

Example 3

Preparation of (Z) -2- [ [3 , 5-bιs (1, 1-dιmethylethyl) -4- hydroxyphenyl]methylene] -4-methyl-3-thiomorpholmone.

A. Preparation of pivaloyl protected 3 , 5-dι-t-butyl-4- hydroxy benzaldehyde

To a stirred solution of 3 , 5-dι-t-butyl-4-hydroxy benzaldehyde (213.2g, 910mmole) dichloromethane (2.73L) was added triethylamine (139.5ml, lmole) . Then a solution of pivaloyl chloride (123.3ml, lmole) in dichloromethane (455ml) was added while the reaction temperature was mamtained at 25°C with an ice bath. After an additional 10mm of stirring the reaction was extracted with 1.8L of water. The organic layer was dried over sodium sulfate, evaporated to dryness, and used without further purification.

B. Preparation of the lithium enolate of 4-methyl-3- thiomorphol one n-Butyl lithium in hexane (135.5ml, 182mmole) was added to 4-methyl-3-thιomorpholmone (23.88g, 182mmole) (prepared as in Example IA) m tetrahydrofuran (546ml) at 0°C over δmmutes. The reaction was allowed to stir for 3 mmutes before the ice bath was removed. After an additional 10 m utes the solution was cooled to -78°C and kept at that temperature until ready for use.

C. Preparation of (Z) -2- [ [3 , 5-bιs (1, 1-dιmethylethyl) -4- hydroxyphenyl]methylene] -4-methyl-3-thiomorpholmone.

To a tetrahydrofuran (1.27L) solution of the protected benzaldehyde, prepared above Step A, which was cooled to -78°C, was added the enolate solution, prepared above m Step B, over 30 mmutes. The reaction was allowed to stir an additional 1 hour at -78°C, then poured mto 910ml water and extracted with ethyl acetate. The organic layer was extracted with brine, dried over sodium sulfate, and evaporated to dryness. The residue was triturated with dichloromethane and filtered to remove recovered 3,5-dι-t- buty1-4-hydroxy benzaldehyde. The filtrate was chromatographed on silica gel usmg a 5-30% acetone in hexane gradient to yield desired product (30.4g, 48%) :

iH NMR (CDCI3) 3 7.9 (s, IH) , 7.5 (s, 2H) , 5.45 (ε, IH) , 3.8

(m, 2H) , 3.2 (s, 3H) , 2.0 (m, 2H) , 1.5 (s, 18H) ; FD MS 347 (M ⁺ ) ;

Elemental Analysis for C20 ^H 29 ^NO 2S Theory : C, 69.12; H, 8.41; N, 4.04. Found: C, 69.41; H, 8.33; N, 3.76.

Example 4

Preparation of (Z) -2- [ [3 , 5-bιs (1, 1-dιmethylethyl) -4- hydroxyphenyl]methylene] -4-methyl-3-thιomorpholmone-1-oxide.

A solution of (Z) -2- [ [3 , -bιs (1, 1-dιmethylethyl) -4- hydroxyphenyl]methylene] -4-methyl-3-thiomorphol one (1.25g, 3.6mmol) dichloromethane (18ml) was cooled under nitrogen to 0°C. A solution of m-chloroperoxybenzoic acid (0.78g, 3.6mmol) dichloromethane (11ml) was then added dropwise over 5 mmutes. The reaction mixture was stirred an

additional 25 m utes and washed once with saturated sodium bicarbonate. The organic layer was dried over sodium sulfate. Rotary evaporation gave crude prodαct which was crystallized from ethyl acetate-hexane to give desired product (1.04g, 79%) : lH NMR (CDCI3) d 8.45 (ε, IH) , 7.65 (s, 2H) , 5.7 (s, IH) , 4.65 (ddd, J=12, 12, 2Hz, IH) , 3.5 (ddd, J=12 , 2, 2Hz, lH) , 3.2 (s, 3H) , 3.15 (m, IH) , 2.95 (ddd J=12, 12, 2Hz, IH) , 1.5 (ε, 18H) ; FD MS 363 (M ⁺ ) ; Elemental Analysis for C20 ^H 29NO3S

Theory: C, 66.08; H, 8.04; N, 3.85. Found: C, 66.18; H, 8.09; N, 3.88.

Example 5

Preparation of (Z) -2- [ [3, 5-bιε (1, 1-dιmethylethyl) -4- hydroxyphenyl]methylene] -4-methyl-3-thιomorpholmone-1, 1- dioxide.

A solution of (Z) -2- [ [3 , 5-bι (1, 1-dιmethylethyl) -4- hydroxyphenyl]methylene] -4-methyl-3-thiomorpholmone (1.25g, 3.6mmol) dichloromethane (18ml) was cooled under nitrogen o to 0 C. A solution of m-chloroperoxybenzoic acid (1.55g, 7.2mmol) dichloromethane (22ml) was then added dropwise over 15 mmutes. The reaction was allowed to stir at room temperature for 20 hours and washed twice with saturated sodium bicarbonate. The organic layer was washed once with brine and dried over sodium sulfate. Rotary evaporation gave crude product which was crystallized from ethyl acetate- hexane, then recrystallized from dichloromethane-hexane to give desired product (1.15g, 84%) as the dichloromethane solvate:

!H NMR (CDCI3) 3 8.4 (s, IH), 7.85 (ε, 2H) , 5.8 (ε, IH) , 5.3 (ε, 2H) , 3.85 (m, 2H) , 3.4 (m, 2H) , 3.2 (ε, 3H) , 1.5 (s, 18H) ; FD MS 379 (M+) ;

Elemental Analysis for C20H29NO4S ^• CH2CI2 : Theory C, 54.55; H, 6.83; N, 3.01. Found: C, 54.30; H, 6.74;N, 3.02.

Example 6

Preparation of (Z) -2- [ [3 , 5-bιε (1, 1-dιmethylethyl) -4- hydroxyphenyl]methylene] -4-ethyl-3-thiomorpholmone.

Title compound was prepared substantially accordmg to the procedure for Example 3 given above: iH NMR (CDCI3) 3 7.84 (s, IH) , 7.47 (ε, 2H) , 5.41 (s, IH) ,

3.77 (m, 2H), 3.59 (q, J=8 Hz, 2H), 2.99 (m, 2H) , 1.46 (s,

18H) , 1.23 (t, J=8 Hz, 3H) ;

FD MS 361 (M ⁺ ) ;

Elemental Analysiε for C21H31NO2S Theory : C, 69.77; H, 8.64; N, 3.87.

Found: C, 70.05, H, 8.60; N, 3.61.

Experimental Autoimmune Encephalomyelitis (EAE) Model Experimental autoimmune encephalomyelitis (EAE) is an inflammatory autoimmune demyelinatmg disease which can be induced m laboratory animals by injection of myelin basic protein. Such disease has become the standard laboratory model for studying clinical and experimental autoimmune diseases. In fact, numerous articles [e.g., Abramsky, e__ ^1. , J. Neuroimmunol . , 2_, 1 (1982) and Bolton e_t a_l. , J.

Neurol. Sci.. 56, 147 (1982)] note that the similarities of chronic relapsing EAE m animals to multiple sclerosis m

humans especially implicates the value of EAE for the study of autoimmune demyelinatmg diseases such as multiple sclerosis. As such, the EAE test model was employed to establish the activity of the compounds of formula I agamst multiple sclerosis. Such testing was conducted accordmg to the following procedure.

Female Lewis rats (Olac Ltd., U.K.), were injected in their footpads with 12.5 mg of myelin basic protein (MBP) (prepared from gumea-pig spinal cord) m Complete Freunds adjuvant. Test compound was given daily from day 0 (MBP injection date) in carboxymethylcelluloεe p.o. at a dosage of 33 mg/kg to the test animals. A control solution (carboxymethylcellulose alone) was given to certam other test animals. The animals were then weighed and scored daily for symptomε of EAE accordmg to a scale of 0 to 3 (0= no change; 1= flaccid tail; 2= hmd limb disability and 3= hmd quarter paralysis/moribund) . Animals were sacrificed when they reached a score of 3.

The results of the experiment described above are set forth Table I, below. In Table I, Column 1 indicates the example number of the test compound employed or, if appropriate, that no test compound was employed (control) . Columns 2-16 report the EAE disease score asεociated with variouε times after the MBP injection date (day 0) .

TABLE I Inhibition of EAE

Compound EAE Disease Score At Various Example Days After MBP Administration *

No./ Control

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Control 0 0 0 0 0 0 0 0 0.13 0.25 1.38 2.75 3 3 3

Example 0 0 0 0 0 0 0 0 0 0.5 0.5 2.2 2. 2.4 2.4 3 4

*EAE Disease Score based on an average of 6 test animals.

The results set forth m Table I, above, establish that, given the proper dosmg paradigm, the compounds of formula I inhibit the progression of EAE. As such, the compounds of formula I would be expected to be efficacious m treating multiple sclerosis.

Pharmaceutical Formulations As noted above, the compoundε of formula I are capable of slowing the process of neurodegeneration associated with multiple sclerosis, thereby lending themselves to the valuable therapeutic method claimed herein. This method comprises administering to a mammal in need of treatment for multiple scleroεis an amount of one or more compounds of formula I sufficient to achieve the therapeutic effect desired. The compounds can be administered by a variety of routes mcludmg the oral, rectal, transdermal, subcutaneous, intravenous, intramuscular or intranasal routes. The oral

and tranεdermal routeε of administration are preferred. No matter what route of administration is chosen, such administration is accomplished by means of pharmaceutical compositions which are prepared by techniqueε well known the pharmaceutical sciences.

In making the pharmaceutical compositions, one or more active ingredients will usually be mixed with a carrier, or diluted by a carrier, or enclosed within a carrier, or diluted by a carrier, or enclosed within a carrier which may be the form of a capsule, sachet, paper or other contamer. When the carrier serves as a diluent, it may be a solid, semi-solid or liquid material which acts as a vehicle, excipient or medium for the active ingredient. Thus, the compositions can be m the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutionε, εyrups, aerosols (as a solid or in a liquid medium) , ointments containing for example up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders.

Some examples of suitable carriers, excipients, and diluentε include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, algmates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, salme solution, syrup, methylcellulose, methyl- and propylhydroxybenzoates, talc, magnesium stearate and mineral oil. The formulations can additionally include lubricating agents, wetting agents, emulsifying and suspending agents, preserving agents, sweetening agents or flavoring agents. The compositions may be formulated so as to provide rapid, sustained or delayed release of the active ingredient after

administration to the patient by employing procedures well known in the art.

The compositions are formulated, preferably in a unit dosage form, such that each dosage contains from about 5 to about 500 mg, more usually about 25 to about 300 mg, of the active ingredient. The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit contammg a predetermined quantity of active material calculated to produce the desired therapeutic effect, aεεociation with one or more suitable pharmaceutical diluents, excipients or carriers.

The compounds utilized m the method of the present invention are effective over a wide dosage range for the treatment of multiple sclerosis. Thus, as used herem, the term "therapeutically effective amount" refers to a dosage range of from about 0.5 to about 50 mg/kg of body weight per day. In the treatment of adult humans, the range of about 1 to about 10 mg/kg, in smgle or divided doseε, is preferred. However, it will be underεtood that the amount of the compound actually administered will be determmed by a physician, m the light of the relevant circumstances including the choice of compound to be administered, the chosen route of administration, the age, weight, and response of the individual patient, and the severity of the patient's symptoms, and therefore the above dosage ranges are not intended to limit the scope of the invention any way. Multiple sclerosis can exist as either an acute or chronic condition. The term "acute" means an exacerbated condition of short course followed by a period of remission. The term "chronic" means a deteriorating condition of slow progress and long continuance. Symptoms are myriad, depending upon the area in the bram where lesions occur, and

can occur anywhere m the body. Symptoms may include such conditions aε weakneεs, blindness, speech difficulties, sensory changes, memory change and εo forth. It is contemplated that the present mvention encompasses the treatment of both acute and chronic forms of multiple sclerosis. In the acute form, compound is administered at the onset of symptoms and discontinued when the symptomε disappear. A chronic condition is treated when it is diagnosed as chronic and continued throughout the course of the disease.

The following formulation examples may employ aε active ingredients any of the compoundε of formula I. The examples are illustrative only and are not intended to limit the scope of the mvention m any way.

Formulation 1

Hard gelatin capsules are prepared usmg the following ingredients : Quantity (mσ/capsule)

2-[ [3,5-bιs(l,l-dιmethylethyl)-4- hydroxyphenyl]methylene] -4-

(2-hydroxyethyl) -3- thiomorpholmone 500 Starch dried 200

Magnesium 10

The above ingredients are mixed and filled mto hard gelatin capsules 710 mg quantities.

Formulation 2

A tablet formula iε prepared using the ingredients below: Quantity (mσ/tablet)

2-[ [3 ,5-bis(l,l-dimethylethyl) -4- hydroxyphenyl]methylene] -4-

[2- (dimethylamino) ethyl] -3-thio orpholinone 100 Cellulose, microcrystalline 400

Silicon dioxide, fumed 10

Stearic acid 5

The components are blended and compresεed to form tablets each weighing 515 mg.

Formulation 3

Tablets each contammg 50 mg of active mgredient are made up as follows:

Quantity (mα/tablet'

2-[ [3,5-bιs(l,l-dιmethylethyl)-3- hydroxyphenyl]methylene] -4- (methylsulfonyl) -3- thiomorpholmone 50 mg

Starch 50 mg

Microcrystalline celluloεe 40 mg

Polyvinylpyrrolidone

(aε 10% solution in water 4 mg Sodium carboxymethyl starch 4.5 mg

Magnesium stearate 0.5 mg

Talc 1 mα

Total 150 mg

The active mgredient, starch and cellulose are passed through a No. 45 mesh U.S. sieve and mixed thoroughly. The solution of polyvinylpyrrolidone is mixed with the resultant powders which are then passed through a No. 14 mesh U.S. sieve. The granules so produced are dried at 50-60 'C and passed through a No. 18 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate and talc, previously passed through a No. 60 mesh U.S. Sieve, are then added to the granules which, after mixing, are compressed by a tablet machme to yield tablets each weighing 150 mg.

Formulation 4

Capsuleε each containing 25 mg of medicament are made as follows:

Quantity (mα/capsule)

2- [ (4-hydroxy-3 , 5-dι-2-propenylphenyl) methylene] -4-methyl-3-thiomorpholmone 25 mg Starch 60 mg

Microcrystalline cellulose 60 mg

Magnesium stearate 5 mq

Total 150 mg

The active ingredient, cellulose, starch and magneεium stearate are blended, passed through a No. 45 mesh U.S. sieve, and filled mto hard gelatin capsules 200 mg quantities.

Formulation 5

Suppositories each containing 250 mg of active ingredient are made up as follows:

Quantity (mσ/suppository)

2- [ (4-hydroxy-3 , 5-dinιtrophenyl)methylene] -4-methyl-3-thiomorpholinone; 250 mg Saturated fatty acid glycerides to 2,000 mg

The active ingredient is passed through a No. 60 mesh U.S. sieve and suεpended in the εaturated fatty acid glycerides previously melted using the minimum heat necessary. The mixture is then poured into a suppository mold of nominal 2 g capacity and allowed to cool.

Formulation 6

Suspensions each contammg 100 mg of medicament per 5 ml dose are made as follows:

Quantity (mα/5mll

2- [ (3 , 5-dichloro-4-hydroxyphenyl)methylene]

- -methyl-3-thiomorpholmone; 100 mg Sodium carboxymethylcellulose 50 mg

Syrup 1.25 ml

Benzoic acid solution 0.10 ml

Flavor q.v.

Color q.v. Purified water to 5 ml

The medicament is passed through a No. 45 mesh U.S. sieve and mixed with the sodium carboxymethyl celluloεe and syrup to form a smooth paste. The benzoic acid solution, flavor and color are diluted with εome of the water and added, with stirring. Sufficient water is then added to produce the required volume.

Formulation 7

Capsules each containing 5 mg of medicament are made up aε follows:

Quantity (mα/tablet!

2- [ ( 3 , 5-dichloro-4-hydroxyphenyl)methylene]

-4-methyl-3-thiomorpholinone; 5 mg Starch 164 mg

Microcrystalline cellulose 164 mg

Magnesium stearate 22 mσ

Total 355 mg

The active ingredient, cellulose, starch and magnesium stearate are blended, passed through a No . 45 mesh U.S. sieve, and filled into hard gelatin capsules in 355 mg quantities.