CA2198114A1 - Methods of inhibiting demyelinating and dysmyelinating diseases - Google Patents

Abstract

A method of inhibiting demyelinating or dysmyelinating diseases or their symptoms comprising administering to a human in need thereof an effective amount of a compound having formula (I) wherein R1 and R3 are independently hydrogen, -CH3, (a) or (b), wherein Ar is optionally substituted phenyl; R2 is selected from the group consisting of pyrrolidine, hexamethyleneimino, and piperidino; or a pharmaceutically acceptable salt of solvate thereof.

Description

W09~0s8~ 2 1 9~ 4 PCT~S95/1058l METHODS OF INHIBITING DEMYE~INATING AWD DESNYE~INATING
nr~ c Diseases of the myelin sheath are divided into two categories. The first category is the demy~lin~ting or myelinoclastic diseases and the second is the dysmyelinating diseases. The myelinoclastic (demyelinating) diseases contain multiple sclerosis, myelinoclastic diffuse sclerosis, post-infectious and postvaccinal encephalomyelitis (disseminated vasculomyelinopathy), tLdll~v~L~e myelitis, central pontine myelinolysis, and marchiafava-bignami disease.
The dysmyelinating diseases (leukodystrophies) contain metachromatic leukodystrophy, globoid cell leukodystrophy, adrenoleukodystrophy, andspongy sclerosis.
The first group (the demyelinating diseases) includes conditions in which the myelin sheath has developed normally and has a completely normal metabolic r-;ntPn~nre system, but in which the sheath appears to be the primary target of various conditions. In this group, two subgroups are recognized. The first one is multiple sclerosis, and some of its variants. The second group consists of the complications of various infections, pr;nrip~lly viral, and vaccinations, which result from a misdirection of the immune response that has been activated by the infection or vaccination. Therefore, both humoral and ce~lular (delayed) immune factors cause the myelin sheath of either the central or the peripheral nervous systems, or of both, to become in~ , edematous, or~0 destroyed.
n the second category, the dysmyelinating diseases, an inborn error of metabolism causes a disturbance of myelinogenesis. The dysmyelination may result from a metabolic failure of the myelin maintenance system after normally formed myelin has been laid down.

W096/058~ 2 1 9 ~: 1 1 4 - PCT~S9~10581 The dysmy~l in~t;ng diseases are of many types and include such disparate conditions as the leukodystrophies the gangliosidoses, such as Tay-Sachs disease; several amino-acidopathies, such as phenylketonuria and maple syrup urine disease; and probably other metabolic disturbances which in one way or another interfere with the normal development of myelin.
It should be pointed out that destruction of myelin is a nonspecific resuIt of almost any injury to the nervous system and can be seén following physical trauma or vascular insults, with infections and neoplasms, or as a result of various intoxications. The conditions considered here are those in which such causes and factors are~not recognized. Two conditions, central pontine myelinolysis and Marchiafava-Bi~nami disease, have been included because they fulfill the criteria of being primary diseases of myelin, although they are not easily classifiable as either variants of multiple sclerosis or instances of -:
postinfectious or postvaccinal reactions of the nervous system.
The conditions to be described here affect primarily, but not exclusively, the central nervous 8ystem.
While multiple sclerosis is a disease restricted to the central nervous system, the leukodystrophies normally also involve the peripheral nerves; the postinfectious or ~-~
postvaccinal reactions may be restricted to the central nervous system ~Pn~ph~ll yelitis), or peripheral nervous system ~ in-Barré syndrome) or may involve both to varying degrees. = ~ . 3 In the experimental allergic encephalomyeIitis AE) animal model, administration of myelin basic protein ~BP) induces EAE ~Higgins et al.~ J. Immunol ., 140 (2), 440-445, January 15, 1988; Bitar et al., Cell. Immun., 112, 364-370, (1988), and is characterized by increased levels of TGF~ and ~L-5 in the brain (Rhoury et al., J. ~xp. Med., 176, 1355-1364, Nov. 1992). Agents which induce TGE~and W096/05823 21~8 ~ ~ 4 r~ ,3~10~ul ~ , other ,3nt;infl, tory cytokine(s) may be useful for the ~c~r;h~ diseases and their symptoms.

This invention provides methods of inhibiting demy~1in~ting or dysmyelinating diseases or their symptoms comprising administering to a human:in need thereof an e~fective amount of a compound of formula I

~ OCH.CH2--R2 R10 ~ oR3 (I) wherein R1 and R3 are independently hydrogen, O O

-C~3 -C-(C1-C6 alkyl), or -C-Ar , wherein Ar is optionally substituted phenyl;

~ R2 is selected from the group consisting of pyrrolidino, hexamethyleneimino, and piperidino; and pharmaceutically acceptable salts and solvates thereof.

The current invention concerns the discovery that a select group of 2-phenyl-3-aroylbenzothiophenes (ben~othiophenes), those of formula I, are useful for inhibiting demyelinating or dysmyelïnating diseases or their symptoms. The compounds of formula I induce the expression of TGF~, and more specifically TGF~-3, and this indicates usefulness for inhibition of the diseases or their symptoms.

2 ~ 98 1 1 4 ~

The therapeutic and prophylactic treatments provided by this invention are practiced by administering to a human in need thereof a aose of a compound of formula I or a r~rr-r~lt;cally acceptable salt or solvate thereof, that is effective to inhibit demyelinating or ~ ~
dysmyelinating diseases or their symptoms or its symptoms.
The term ~inhibit~' includes its~generally accepted meaning which includes prohibiting, preventing, restraining, and slowing, stopping or reversing progression, severity or a resultant symptom. As such, the present method includes both medical therapeutic andfor prophylactic administration, as appropriate.
Raloxifene is a preferred compound of this invention and it is the hydrochloride salt of a compound of formula l wherein Rl and R3 are hydrogen and R2 is 1-piperidinyl.
Generally, at least one compound of formula I
is formulated with common excipients, diluents or carriers, and compressed into tablets, or formulated as elixirs or solutions for convenient oral administration, or administered by the intramuscular or intravenous routes.
The compounds can be administered transdermally, and may be formulated as sustained release dosage forms and the Iike.
The compounds used in the methods of the current invention can be made accordi~g to established procedures, such as those detailed i~ U.S. Patent Nos. 4,133,814, 4,418,068, and 4,380,635 all of which are incorporated by reference herein. In general, the process starts with a ben~o[b]thiophene having a 6-h~ydroxyl group and a 2-(4-hydroxyphenyl~ group The starting compound is protected,acylated, and deprotected to form the formula I compounds.
Examples of the preparation of such compounds are provided in the U.S. patents discussed above. The term "optionally substituted phenyl" includes phenyl-ana phenyl substituted once or twice with Cl-C6 alkyl~, Cl-C4 alkoxy, hydroxy, nitro, chloro, fluoro, or tri~chloro or fluoro)methyl.

~ W096/058~ 2 ~ 9 8 1 ~ 4 PCT~S9~10581 The compounds used in the methods of this invention form pharmaceutically acceptable acid and base addition salts with a wide variety of organic and inorganic acids and bases and include the physiologically acceptable salts which are often used in pharmaceutical chemistry.
Such salts are also part of this invention. Typical inorganic acids used to form such salts include hydrochloric, hydrobromic, hydroiodic, nitric, sulfuric, phosphoric, hypophosphoric and the like. Salts derived from organic acids, such as ~1iph~t;c mono and dicarboxylic acids, phenyl substituted alkanoic acids, hydroxyalkanoic and hydroxyalkandioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, may also be used. Such pharmaceutically acceptable salts thus include acetate, phenylacetate, trifluoroacetate, acrylate, ascorbate, ben_oate, chlorobenzoate, dinitrnhpn7o~te~ hydroxybenzoate, methoxybenzoate, methylbenzoate, o-acetoxybenzoate, naphthalene-2-benzoate, bromide, isobutyrate, phenylbutyrate, ~-hydroxybutyrate, butyne-1,4-dioate, hexyne-1,4-dioate, caprate, caprylate, chloride, cinnamate, citrate, formate, fumarate, glycollate, heptanoate, hippurate, lactate, malate, maleate, hydroxymaleate, malonate, mandelate, mesylate, nicotinate, isonicotinate, nitrate, oxalate, phth~l~te~ teraphthalate, phosphate, monohydrogenphosphate, dih~dlu~e-lullosphate, met~hncph~te, pyrophosphate, propiolate, propionate, phenylpropionate, salicylate, sebacate, succinate, suberate, sulfate, bisulfate, pyrosulfate, sulfite, bisulfite, sulfonate, benzene-sulfonate, p-bromophenylsulfonate, chlorobPn7~n~c~lifonate, ethanesulfonate, 2-hydroxyethanesulfonate, meth~neql-1fonate, naphthalene-1-sulfonate, n~phth~l ene-2-sulfonate, p-toluenesulfonate, xylenesulfonate, tartarate, and the like. A preferred salt is the hydrochloride salt.
= . The pharmaceutically acceptable acid addition salts are typically formed by reacti~g a compound of 2~:981~
W09610S8~ PCTNS95/10581 formula I with an e~uimolar or excess amo~nt of acid. The reactants are gene~ally ~ d in a mutual solvent such as diethyl ether or benzene. ~he salt normally precipitates o~t of solution within about one hour to= lO
days and can be isolated by filtration~or the solvent can be stripped off:by conventional means.
sases commonly used for formation of salts include ammonium hydroxide an=d alkali and alkaline earth metal hydroxides, carbonates, as well as aliphatic and primary, secondary and tertiary amines, aliphatic diamines Bases especially useful in the preparation of addition salts include ammonium hydroxide, potassium carbonate, methylamine, diethylamine, ethylene diamine and cyclohexylamine ~he pharmaceutically acceptable salts generally have enhanced solubility characteristics compared to the compound from which they are derived, and thus are often more ~r-nAh1~ to formulation as liguidc or emulsions.~
Pharmaceutical formulations can be ~repared by procedures known in the art. For example, the compounds can be formulated with common excipients, diluents, or carriers, and formed into tablets, capsules, suspensions, powders, and the like. Examp:les of eXcipients, diluents, and carrier-s that are c-1it~h1~ for such formuIations include the following: fillers and ex~ënaers such as starch, sugars, mannitol, and silicic derivatives; binding agents such as carboxymethyl cellulose and other cell~lose derivativec, alginates, gelatin, and polyvinyl pyrrolidone;
moisturi~ing agents such as glycerol; disintegrating=agents such as calcium carbonate and sodium bicarbonate; agents for retarding dissolution such as paraffin; resorption accelerators such as guaternary ;nm com~oounds; surface active agents such as cetyl alcohol, glycerol monostearate;
adsorptive carriers such as kaolin and bentonite; and lubricants such as talc, calcium and r-~n~c;~1m stearate, and solid polyethyl glycols.~

~ W09~05823 2 ~ 4 r~l/U~ ~ i The compounds can also be formulated as elixirs or solutions for convenient oral administration or as solutions appropriate for parenteral administration, for instance by intramuscular, subcutaneous or intravenous routes. Additionally, the compounds are well suited to formulation as sustained release dosage forms and the like.
The formulations can be so constituted that they release the active ing~edient only or preferably in a particular part of the intestinal tract, possibly over a period of time. The coatings, envelopes, and protective matrices may be made, for example, from polymeric substances or waxes.
The particular dosage of a compound of formula I
re~uired to inhibit demyelinating or dysmyelinating diseases or their symptoms according to this invention, will depend upon the severity of the condition, the route of administration, and related factors that will be decided by the attending physician. Generally, accepted and effective daily doses will be from about 0.1 to about 1000 mg/day, and more typically from about 50 to about 200 mg/day. Sùch dosages will be administered to a subject in need thereof from once to about three times each day, or more often as needed, and for a duration to effectively inhibit the disease(s) or symptom(s~.
It is usually preferred to administer a compound of formula I in the form of an acid addition salt, as is customary in the administration of pharmaceuticals bearing a basic yroup, such as the piperidino ring. For such purposes the following oral dosage forms are available.

Formulatinnq In the fo l~tinn~ which follow, "Active ingredient" means a compound of formula I.

2'1.~114 ~
W096/OS8~ PCTNS95110S81 F llstion 1: Gelatin Capsules Hard gelatin capsules are prepared using the following:

IngredientQuantlty (mq~c~psule) Active ingredient 0.1 - 1000 StArch, NF O - 650 StArch flow~ble powder0 - 650 Silicone fluid 350=centistokes 0 - 15 The ingredients are blended, passed through a No. 45 mesh U.S. sieve, and filled int~o hard gelatin capsules.~~--Examples of specific capsule~formulations ofraloxifene that have been made include those shown below:

For~nl~tion 2: Raloxifene capsule IngredientQu~ntity (mg/cApsule) Raloxifene St~rch, NF 112 StArch flow~ble powder 225.3 Sillcone fluid 350 centistokes 1.7 F 1l ~tion 3: Raloxifene capsule IngredientQu~ntitv (mg/c~psule) R~loxifene 5 StArch, NF 108 StArch flowAble powder 225.3 Silicone fluid 350 centistokes 1 7 ~ W096/oS8~ 21 ~8 ~ 14 r~ c~ ' Fo lAtion 4: Raloxifene capsule -Ingredient Quantity (mq/capsule) Raloxifene 10 starch, NF 103 Starch flow~ble powder- 225.3 Silicone fluid 350 centistokes 1.7 Formn7~t;on 5: Raloxifene capsule ~
Ingredient = = Quantity (mqtcapsule) Raloxifene 50 Starch, NF 150 StArch flowable powder 397 Silicone fluid 354 centistokes 3.0 The specific f( llatinn~ above may be changed in rn~ nre with the reasonable variations provided.
A tablet formulation is prepared using the ingredients below:

For~n~tion 6: Tablets Inqredient Quantity (mq~tabietj Active ingredient 0.1 - 1000 Cellulose, microcrystalIine 0 - 650 Silicon dioxide, fumed0 - 650 Ste~rate acid 0 - 15 The components are blended and compressed to form tablets.
Alternatively, tablets each cnnt~ining 0.1 -lO00 mg of ActIve ingredient are made up as follows:

21~811~ ~
W096/05823 ~ PC~NS95/10581 F ~l~tion 7: Tablets IngredientQu~ntity tmg2t~blet) Active ingredient 0.1 - 1000 =~ = r St~rch 95 Cellulose, microcrystalline 35' Polyvinylpyrrolidone 4 (~s 10~ solution in waterl Sodium c~rboxymethyl cellulose ~ = ~.5 Magnesium ste~r~te 0.5 T~lc 7 The Active ingredient, starch, and cellulose are passed through a No. 45 mesh U.S. sieve and mixad 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, ~gn~ m stearate, and talc, previously passed through a No. 60 U.S. sieve, are then added to the granules which, after. mixing, are ~-compressed on a tablet machine to yield tablets.
Suspensions each containing 0 1 - 1000 mg of Active ingredient per 5 m~ dose are made as follows:

Formulation ~: Suspensions Ingredient Qu~ntity ~mq/5 mll Active ingredient 0.1 - 1000 mg Sodium c~rboxymethyl cellulose : 50 mg Syrup 1.25 mg Benzoic ~cid solution O.10 mL
Fl~vor ~.= q.v.
Color q.v.
Purified w~ter to S mL

~ W096/OS8~ 2 t 98 1 t 4 PCT~SgS/10581 The Active ingredient is passed through a No. 45 mesh U.S.
sieve and mixed with the sodium carboxymethyl cellulose and syrup to form a smooth paste. The benzoic acid solution, ~lavor, and color are diluted with some of the water and added, with stirring. Sufficient water is then added to produce the required volume.

ASSAYS
Assav 1 Experimental allergic encephalomyelitis ~EAE1 is an acute ;nf~. ~tory autoimmune disease of the central nervous system that can be elicited in rodents and is the major animal model for the study of multiple sclerosis (MS). EAE is a systemically initiated autoimmune disease in which spinal cord homogenate or myelin basic protein (MsP) prepared in suitable adjuvants, such as complete Freundls adjuvant (CFA), is injected to activate the peripheral immune system. Rapid migration of activated T
cells from peripheral blood to the CNS then occurs where they initiate a localized infl. tinn and subsequent demyelination. A monophasic, acute or spontaneous relapsing-remitting chronic form of the disease may develop - according to the mode of sensiti~ation, genetic background and age of the animal. Alternatively, the chronic relapsing form of EAE can also be induced by injection of MBP-specific T=cell lines or clones of the helperjinducer phenotype (CD4+). The resulting demyrlin~tinn closely resembles that obtained by injection of MBP.
Typically~ spinal cord homogenate or M3P is mixed with CFA at a 1:1 ratio. Rodents are injected s.c.
in the posterior flank at multiple sites with not less than 1 mg sensitizing protein in a final volume of approximately 100 ul. Subsequent inorul~t;nnc~ similar to the primary, can be administered at 7 day intervals. Rodents usually demonstrate severe neurological symptoms as early as 1~
days after primary inoclTl~t;nn with peak incidence levels , .. . . . . . . _ .. .. . _ ,, .. . . . , , _ _ W096/058~ 2 1 ~ 1 1 4 of EAE=being reached by 21 days. S~ t~n~us injection of as few as lOe5 ~BP-specific T~cell line or clone cells of the CD4+ helperlinducer phenotype is also~effective at inducing EAE. Compound is administered orally or s.c., before or aiter2primary antig-en inoculation, and the beneficial effects determinea by evaluation of~ neurological symptoms associated with EAE disease progression. O~e can also examine cerebrospinal fluid or brain ~or rhe presence of i~fl, ~tory ceIIs, oligo~lonal IgG or incr~ased class II expression.

~ .
Five to fifty patients are selected for the clinical study. The patients-suffer from a demyelinating or desmyelinating disease~which exhibi~s symptoms, but otherwise are ~n good general health. Because of the idiosyncratic and subjective nature of these disorders, the study has a placebo control group, i.e., the patients are divided into two groups, one of which receives a comp~ound of iormula 1 as the active agent and the other receives a placebo. Patients in the te~t sroup receive between 50-200 mg of the drug per day by the oral route. ~hey continue this therapy for 3-12 months. Accurate records are kept as to the number and severity of the symptoms in both groups and at the end of the study these results are compared.
The results are compared both between members of each group and also the results for each patient are compared to~ the symptoms report~d by each patient before the study began.

U~ility of the compounds of formula I is illustrated by the positive impact they have in at least one o_ the assays described above _ - ~

Claims (5)

I claim:

1. A method of inhibiting demyelinating or dysmyelinating diseases or their symptoms comprising administering to a human in need thereof an effective amount of a compound having the formula wherein R1 and R3 are independently hydrogen, -CH3, , , wherein Ar is optionally substituted phenyl;
R2 is selected from the group consisting of pyrrolidine, hexamethyleneimino, and piperidino; or a pharmaceutically acceptable salt of solvate thereof.

2. The method of Claim 1 wherein said compound is the hydrochloride salt thereof.

3. The method of Claim 1 wherein said administration is prophylactic.

4. The method of Claim 1 wherein said compound is or its hydrochloride salt.

5. The method of Claim 1 wherein said disease is multiple sclerosis.