RS55903B1 - Obujmica za crevo - Google Patents

Description

DERIVATI FUMARNEKISELINE^KAONF-KAPPAB INHIBITORI

Ovaj pronalazak se odnosi na upotrebu jednog ili više derivata fumarne kiseline kao NF-kappaB inhibitora. U isto vreme, ovaj pronalazak se odnosi na upotrebu derivata fumarne kiseline za pripremanje farmaceutskog sastava za lečenje bolesti na koje se može uticati sa NF-kappaB.

STANJE TEHNIKE

Poznato je da farmaceutski sastavi kao što je fumarna kiselina koja, nakon biološke degradacije posle administriranja, ulaze u ciklus limunove kiseline ili su njegov deo pa odatle dobijaju povećani terapeutski značaj - posebno kada su dati u visokim dozama - pošto mogu da ublaže ili izleče bolest izazvanu kriptogenetički. Dodatno, fumarna kiselina inhibira rast Ehrlich trbušno vodenih tumora kod miševa, redukuje toksična dejstva mitomicina C

i aflatoksina i pokazuje anti-psorijatičku i anti-mikrobnu aktivnost.

Najvažnija praktična primena u lečenju psorijaze sa različitim derivatima fumarne kiseline je već opisana u brojnim patentima, na primer EP 0 188 479, DE 25 30 372, DE 26 21 214 ili EP 0 312 697.

Sledeća upotreba derivata fumarne kiseline, naime alkilhidrofumarata, je objavljena u DE 197 21 099.6 i DE 198 53 487.6 prema kojima su ovi specifični derivati fumarne kiseline opisani za lečenje auto-imunih bolesti kao što je poliartritis, multipleks skleroza i reakcija presada-spram-domaćina. Dodatno, DE 198 53 487.6 i DE 198 39566.3 uče o upotrebi alkilhidrofumarata i dialkilfumarata u transplatnatcionoj medicini, lako su obavljena individualna istraživanja mehanizma dejstva derivata fumarne kiseline u lečenju psorijaze, ne postoji specifična informacija o ovoj temi.

NF-kappaB (nuklearni faktor kappaB) je faktor transkripcije eukariotnih ćelija. NF-kappaB pripada familiji Rel proteina, klasi transkripcionih faktora koje karakteriše takozvani Rel domen. Rel domen je imenovan pošto je prvi član otkriven u avian virusu kao onkogen. Specifični položaji ovog homologog Rel domena (Rel homologi domen = RHD) koji se sastoji od 300 amino kiselina su odgovorni za DNK vezivanje na kappaB položaje, dimerizaciju sa drugim proteinima Rel familije i interakciju sa l-kappaB.

Do sada, kod sisara je poznato pet članova Rel familije. To su c-Rel, NF-kappaBI (p105/p50), NF-kappaB2 (p100/p52) i ReIB. Teorijski, ovih pet članova Rel familije proteina mogu da se kombinuju u bilo koji oblik homo- i heterodimera, iako je samo nekoliko specifičnih kombinacija primećenoin vivo.Klasično i najbolje karakterisan NF-kappaB molekul je heterodimer p50/p65 pod-jedinica NF-kappaB1/RelA. Ovaj heterodimer je najčešći kompleks i pronađen je praktično u svim ćelijskim tipovima.

Posle ćelijske aktivacije i disocijacije l-kappaB, NF-kappaB heterodimer p50/p65 migrira u ćelijski nukleus gde se vezuje za saglasnu sekvencu 5'-GGGRNNYYCC-3'. U ovom postupku, p50 pod-jedinica primarno služi kao pod-jedinica za vezivanje DNK, dok p65 pod-jedinica obezbeđuje transaktivacionu funkciju.

Kao rezultat različitih kombinacija, svaki od ovih heterodimera pokazuje jedinstvene karakteristike što se tiče specifičnosti ćelijskog tipa, preferenci što se tiče DNK-vezivanja, različitu interakciju sa 1-kappaB izooblicima, različite zahteve aktivacije i kinetike aktivacije.

Brza izvodljivost NF-kappaB je pripisana činjenici da je faktor prisutan u citoplazmi u neaktivnom obliku, naime kompleks vezan za NF-kappaB inhibitor 1-kappaB. Prema tome, nova sinteza proteina za aktivaciju se ne zahteva, već samo rastvaranje ovog kompleksa sa 1-kappaB ili degradacija ovog inhibitora i potonja translokacija novog aktivnog NF-kappaB dimera u jezgro.

NF-kappaB se može aktivirati vrlo različitim fiziološkim i ne-fizioliškim nadražajima. Ovo uključuju citokine, mitogene, viralne proizvode, unakrsno-povezivanje antigen receptora na T- i B-limfocitima, kalcijum jonofore, forbol estre, UV-zrake, oksidacioni pritisak, inhibitore fosfataze i drugo. Raspon mnogih NF-kappaB regulisanih ili aktiviranih gena je jednako širok, transkripcija kojom je aktiviran, izazvana ili pojačana vezivanjem heterodimera na saglasnu sekvencu kako je to gore opisano. Posebno TNF-alfa, IL-1, IL-2 i lipopolisaharidi mogu biti pomenuti kao važni stimulanti.

Regulisani geni generalno obuhvataju gene koje su uključeni u imunu funkciju, odgovor na inflamaciju, ćelijsku adheziju, ćelijski rast, ali takođe i u ćelijsku smrt. Treba da se posebno pomenu geni molekula ćelijske adhezije, citokini, receptori citokina, proteini akutne faze, faktori rasta i virusni geni. Specijalni geni među njima indukovani sa NF-kappaB su geni za interferon-p, za svetlosni lanac imunoglobulina, za receptor T-ćelije, za TNF-a i TNF-(3 i za faktor tkiva (CD142), ranije nazvan tromboplastin tkiva ili faktor III.

Dugujući svojoj gore pokazanoj centralnoj ulozi u regulaciji imunih reakcija i odgovora na inflamaciju i svom uključivanju u regulaciju faktora tkiva, citokina itd., pretpostavlja se da prednosti slične onim koje su već poznate iz anti-inflamatornih sredstava mogu biti očekivane iz razvoja selektivnih inhibitora za transkripciju faktora NF-kappaB. Steroidna anti-inflamatorna sredstva, interferoni i ciklosporin mogu biti imenovani kao primeri.

OPIS PRONALASKA

Iznenađujuće, sada je otkriveno da individualni derivati fumarne kiseline ili njihove smeše imaju NF-kappaB inhibirajuće dejstvo. Pretpostavljeno ovo dejstvo može biti upotrebljeno za pripremanje farmaceutskog sastava koji sadrži individualno ove derivate fumarne kiseline ili u primesi za lečenje bolesti koje su posredovane ili na koje se može uticati sa NF-kappaB. Posebno, bolesti na koje se može uticati sa NF-kappaB su progresivna sistemska sklerodermija, osteochondritis svphilitica (Wegener-ova bolest),

cutis marmorata (livedo reticularis), Behcet-ova bolest, panarteriitis,

ulcerozni kolitis, vaskulitis, osteoartritis, podagra, aretrioskleroza, Reiter-ova bolest, pulmonalna granulomatoza, tipovi encefalitisa, endotoksični šok (septični/toksični šok), sepsa, pneumonija, encefalomijelitis, anorexia nervosa, hepatitis (akutni hepatitis, hronični hepatitis, toksični hepatitis,

alkoholom izazvani hepatitis, virusni hepatitis, žutica, insuficijencija jetre i citomegalovirusni hepatitis), RennertT-limfomatoza, mesangial nefritis, post-

angioplastična restenoza, reperfuzioni sindrom, citomegavirusna retinopatija, adenovirusne bolesti kao što su adenovirusni nazebi, adenovirusna faringokonjuktivistična groznica i adenovirusna oftalmija, AIDS, Guillain-Barre sindrom, post-herpetična ili post-zoster neuralgia,

inflamatorna demijelizirajuća polineuropatija, mononeuropatija multipleks,

muskoviscidoza, Bechterew-ova bolest, Barett-ov ezofagus, EBV (Epstein-Barr virusna) infekcija, srčano remodelovanje, intersticijalni cistitis, dijabetis mellitus tip II, humani radioaktivno senzibilni tumor, multi-otpornost malignih ćelija na hemoterapeutska sredstva (otpornost mulitleka u hemoterapiji),

granualoma annulare i kanceri kao što su karcinom dojke, karcinom debelog creva, melanom, primarni karcinom ćelije jetre, adenokarcinom, Kapošijev sarkom, karcinom prostate, leukemija kao što je akutna mijeloidna leukemija, višestruki mijelom (plasmocvtoma), Burkitt-ov limfosarkom i Castelman-ov tumor.

U skladu sa pronalaskom, jedan ili više derivata fumarne kiseline odabran od grupe koja se sastoji od dialkil estra fumarne kiseline i monoalkilnih estara fumarne kiseline u obliku sloboden kiseline ili u obliku soli i njihovih smeša se pretpostavljeno upotrebljavaju za NF-kappaB inhibiciju i za pripremanje farmaceutskog sastava.

Dialkil estri fumarne kiseline pretpostavljeno odgovaraju formuli

pri čemuR:i R2koji mogu biti isti ili različiti, nezavisno predstavljaju linearni, račvasti, ciklični, zasićeni ili nezasićeni Ci?4 alkil radikal ili C5_2oari I redikal i ovi radikali su opciono supstituisani sa halogenom (F, Cl, Br, J), hidroksi, Ci_

4alkoksi, nitro ili cijano.

RadikaliR^i R2su pretpostavljeno metil, etil, n-propil, izopropil, n-butil, sek-butil, t-butil, pentil, ciklopentil, 2-etilheksil, heksil, cikloheksil, heptil, cikloheptil, oktil, vinil, alil, 2-hidroksietil, 2- ili 3-hodroksipropil, 2,3-dihidroksipropil, 2-metoksietil, metoksimetil ili 2- ili 3-metoksipropil.

Monoalkil estri fumarne kiseline pretpostavljeno odgovaraju formuli

pri čemu jeR^kako je to gore definisano, A je vodonik, alkali ili alkalni katjon zemnog metala ili fiziološki kompatibilni katjon prelaznog metala, pretpostavljeno odabran od Li<+>, Na<+>, K<+>, Mg<2+>,Ca<2+>, Zn<2+>,Fe<2>+ i Mn<2+>, a n je 1 ili 2 i odgovara valenci A.

Pronalazak pretpostavljeno upotrebljava jedan ili više derivata fumarne kiseline odabranih od grupe koja obuhvata dimetil estar fumarne kiseline, dietil estar fumarne kiseline, metiletil estar fumarne kiseline, metilfumarna kiselina, etilfumarna kiselina, magnezijummetilfumarat,

magnezijumetilfumarat, cinkmetilfumarat, cinketilfumarat,

gvožđemetilfumarat, gvožđeetilfumarat, kalcijummetilfumarat i/ili kalcijumetilfumarat.

U skladu sa pronalaskom, derivati fumarne kiseline za pripremanje farmaceutskog sastava se pretpostavljeno upotrebljavaju u takvoj količini da jedna jedinična doza navedenog farmaceutskog sastava sadrži količinu derivata fumarne kiseline koja odgovara ili je ekvivalentna sa 1 do 500 mg,

poželjno 10 do 300 mg a najpoželjnije 10 do 200 mg fumarne kiseline.

Pretpostavljeni oblici administracije za farmaceutski sastav su oralni, parenteralni, rektalni preko kože, dermalni, nazalni, pulmonalni (inhalacija) ili oftalni (u obliku kapi za oči), pri čemu je poželjna oralna administracija. Sastav će biti prisutan u pogodnom obliku za svaki tip administracije.

Kada se administrira oralno, farmaceutski sastav je prisutan u obliku tableta jediniče doze, mikro-tableta (višestruke jedinične doze tableta) ili mini-tableta, mikro-pilula ili granulata (navedene mikro-tablete, mikro-pilule ili granulat su opciono kapsulirani ili napunjeni u kesice), kapsula ili rastvora za piće. U pretpostavljenoj realizaciji, čvrsta doza ili oblici administracije su obezbeđeni putem enteričnog oblaganja. Takvo oblaganje može takođe biti obezbeđeno na kapsuliranim ili punjenim oblicima doza.

U slučaju parenteralne administracije ubrizgavanjem (i.v., i.m., p.k., Lp.),

sastav je prisutan u pogodnom obliku. Mogu se upotrebiti svi uobičajeni nosači tečnosti pogodni za ubrizgavanje.

Farmaceutski sastav može pretpostavljeno sadržati bilo individualno ili u primesi: 10 do 500 mg dialkilfumarata, posebno dimetilfumarata i/ili dietilfumarata; 10 do 500 mg kalcijumalkilfumarata, posebno kalcijummetilfumarata i/ili kalcijumetilfumarata; 0 do 250 mg cinkalkilfumarata, posebno cinkmetilfumarata i/ili cinketilfumarata; 0 do 250 mg alkilhidrofumarata, posebno metilhidrofumarata i/ili etilhidrofumarata; i 0 do 250 mg magnezijumalkilfumarata, posebno magnezijummetilfumarata i/ili magnezijumetilfumarata, a ukupne gore pomenute količine odgovaraju ekvivalentu od 10 do 500 mg, poželjno 10 do 300 mg a najpoželjnije 100 mg fumarne kiseline.

Pretpostavljeni sastavi pronalaska sadrže samo dimetilfumarat u količini od 10 do 300 mg.

Prema posebno pretpostavljenoj realizaciji, sastav je prisutan u obliku mikro-tableta ili mikro-pilula. Oni pretpostavljeno imaju veličinu ili srednju vrednost prečnika od <5000 (.im, još poželjnije 300 do 2500fim, posebno 300 do 1000 ?m za pilule i 1000 do 2500 |am za mikro-tablete. Administriranjem derivata fumarne kiseline u obliku mikro-tableta, što je pretpostavljeno prema pronalasku, gastro-intestinalne iritacije ili propratni efekti koji ne mogu biti isključeni u slučaju administracije uobičajenih tablete jedinične doze mogu biti dalje smanjeni. Ovo je verovatno zbog činjenice da su mikro-tablete, pretpostavljeno mikro-tablete sa enteričnom oblogom, beć rasute u stomaku pa prema tome dosežu do intestinalnog trakta u porcijama gde se aktivni sastojci oslobađaju u manjim dozama dok ukupna doza ostaje ista. Ovo, za uzvrat, pomaže izbegavanju lokalne iritacije epitelnih intestinalnih ćelija što rezultira poboljšanom gastrointestinalnom tolerancijom mikro-tableta u poređenju sa konvencionalnim tabletama.

Na primer, derivati fumarne kiseline koji se sadrže u sastavima pronalaska su pripremljeni postupkom opisanim u EP 0 312 679.

Primeri proizvodnje

U principu, oralni sastavi pronalaska u obliku tableta ili mikro-tableta mogu biti pripremljeni klasičnim postupcima pravljenja tableta. Umesto takvih klasičnih postupaka pravljenja tableta mogu se upotrebiti drugi postupci za pripremanje tableta, kao što je direktno pravljenje tableta kao i postupci za pripremanje čvrstih disperzija prema postupku topljenja ili postupak sušenja raspršivanjem.

Tablete mogu biti obezbeđene sa enteričnom oblogom. Enterična obloga može se staviti u klasičan sud za oblaganje ili raspršiti. Obloga može takođe biti štavljenja putem Boegel aparata za oblaganje. Dodatno, tableta može biti obezbeđena sa oblogom od filma.

Da bi se objasnila upotreba prema ovom pronalasku, u daljem tekstu dati su različiti primeri za pripremanje pretpostavljenih lekova. Ovi primeri imaju nameru samo da ilustruju ali ne i da limitiraju pronalazak.

Primer 1

Pripremanje film tableta sa enteričnim oblaganjem koje sadrže 100, 0 mg

monometilfumarat- Casoli, koja odgovara 78 mg fumarne kiseline

Preduzimanjem neophodnih predostrožnosti (maska za disanje, zaštitno odelo, itd.), 10 kg monometilfumarat-Ca soli se zdrobi, intenzivno meša i homogenizuje sita 800. Potom se priprema smeša inertnih punilaca sledećeg sastava: 21 kg derivata škroba (STA-RX 1500®), 2 kg mikro-kristalne celuloze (Avicel PH 101®), 0,6 kg polivinil pirolidona (PVP, Kollidon® 25), 4 kg Primogel®, 0,3 kg koloidne silicijumove kiseline (Aerosil®).

Aktivni sastojak se dodaje ukupnoj smeši u obliku pudera, meša se, homogenizuje sa sitom 200 i postupa se sa 2% vodenog rastvora polivinil pirolidona (PVP, Kollidon® 25) na uobičajeni način u vezivne granule, i onda meša sa vanjskom fazom u suvom obliku. Potonje se sastoji od 2 kg takozvanog FST kompleksa koji sadrži 80% talka, 10% silicijumovu kiselinu i 10% magnezijum stearata.

Potom se smesa presuje u konveksne tablete težine od 400 mg i prečnika od 10,0 mm uobičajenim postupkom. Umesto ovih klasičnih postupaka spajanja mogu se upotrebiti za pripremanje tableta drugi metodi kao što je direktna kompresija ili čvrsto raspršivanje prema postupku topljenja i sušenja raspršivanjem.

Enterično oblaganje:

Rastvor 2,250 kg hidroksipropilmetilcelulozeftalata (HPMCP, Pharmacoat HP® 50) se rastvara u smeši rastvarača koj9i se sastoji od 2,50 litara deminaralizovane vode, 1,3 litra acetona Ph. Helv. VII i 13 litara etanola (94% po težini) i potom se dodaje rastvoru 0,240 ricinusovog ulja (Ph. Eur. II). Rastvor se sipa ili rasprši u delovima na jezgra tableta u posudi za oblaganje na konvencionalan način ili se stavlja putem aparata sa fluidizacionom kolonom odgovarajuće strukture.

Posle sušenja, stavlja se obloga od filma. Navedena obloga se sastoji od rastvora Eudragit E 12,5%® 4,8 kg, talka Ph. Eur. II 0,34 kg, titanijum(VI)oksida Cronus RN 56® 0,52 kg, obojenog laka ZLT-2 plavo (Siegle) 0,21 kg, i polietilen glikola 6000 Ph. Helv. VII 0,12 kg u smeši rastvarača od 8,2 kg 2-propanol Ph. Helv. VII, 0,06 kg glicerin triacetata (Triacetin®) i 0,2 kg demineralizovane vode. Posle homogene distribucije u posudu za oblaganje na fluidizacionu kolonu, smeša se osuši i polira na uobičajeni način.

Primer 2

Pripremanje enteričnog oblaganja kapsula koje sadrže 86, 5 mg

monoetilfumarat- Ca soli i 110, 0 mg dimetilfumarata, koji odgovara ukupno

150 mg fumarne kiseline

Preduzimajući neophodne mere predostrožnosti (maska za disanje, rukavice, zaštitna odeća, itd.), 8,65 kg monoetilfumarat-Ca soli i 11 kg dimetilfumarata se intenzivno mešaju sa smešom koja se sastoji od 15 kg škroba, 6 kg laktoze Ph. Helv. VII, 2 kg mikro-kristalne celuloze (Avicel®), 1 gd polivinil pirolidona (Kollidon® 25) i 4 kg Primogel® i homogenizuje putem sita 800.

Zajedno sa 2% vodenog rastvora polivinil pirolidona (Kollidon® 25) sa ukupnom puder smešom se postupa na uobičajeni način u vezivnom granulatu i meša sa spoljnom fazom u suvom obliku. Navedena spoljna faza se sastoji od 0,35 kg koloidne silicijumove kiseline (Aerosil®), 0,5 kg Mg stearata i 1,5 kg talka Ph.Helv. VII. Homogena smeša se onda puni u porcije od 500,0 mg u odgovarajuće kapsule koje se onda obezbeđuju sa enteričnim (otpornim na želudačnu kiselinu) oblaganjem koje se sastoji od hidroksipropiletilcelulozastearata i ricinusovog ulja kao sredstva za omekšavanje putem poznatih postupaka. Umesto kapsula od tvrdog želatina, smeša se takođe može puniti u odgovarajuće na želudačnu kiselinu otporne kapsule, koje se sastoje od smeše celuloza acetat ftalat (CAP) i hidroksipropiletilcelulozaftalata (HPMCP).

Primer 3

Pripremanje enterično- obloženih mikro- tableta u kapsule koje sadrže 87, 0

mg meonoetilfumarat- Ca soli, 120 mg dimetilfumarata, 5, 0 mg

monoetilfumarat- Mg soli i 3, 0 mg monoetilfumarat- Zn soli, koje odgovaraju

ukupno 164 mg fumarne kiseline(" forte "tablete).

Preduzimanjem neophodnih mera predostrožnosti (maska za disanje,

rukavice, zaštitna odeća, itd.9, 8,7 kg 8,7 kg monoetilfumarat-Ca soli, 12 kg dimetilfumarata, 0,5 kg monoetilfumarat-Mg soli i 0,3 kg monoetilfumarat-Zn soli se drobe intenzivno mešaju i homogenizuju kroz sito 800. Onda se priprema smeša inertnih punilaca sledećeg sastava: 18 kg derivata škroba (STA-RX 1500), 0,3 kg mikro-kristalne celuloze (Avicel PH 101), 0,75 kg PVP (Kollidon 120), 4 kg Primogel-a, 0,25 kg koloidne silicijumove kiseline (Aerosil). Ukupna puder smeša se dodaje smeši aktivnog sastojka, homogenizuje putem 200 sita, i postupa se na uobičajeni način sa 2% vodenog rastvora polivinil pirolidona (Kollidon K25) da bi se dobio vezivni granulat i meša se sa spoljnom fazom koja se sastoji od 0,5 kg magnezijum stearata i 1,5 kg talka. Onda se puder smeša presuje konvencionalnim postupkom u konveksne mikro-tablete sa bruto težinom od 10,0 mg i prečnikom od 2,0 mm. Umesto klasičnog postupka pravljenja tableta mogu se upotrebiti takođe drugi postupci za pravljenje tableta kao što je direktno pravljenje tableta ili čvrste disperzije putem postupka topljenja i postupak suvog raspršivanja.

Oblaganje otporno na želudačnu kiselinu može se sipati ili raspršiti u klasičnoj posudi za oblaganje ili staviti na aparat sa fluidizacionom kolonom. Da bi se postigla otpornost na želudačnu kiselinu, porcije rastvora od 2,250 kg hidroksipropilmetilcelulozaftalata (HPMCP, Pharmacoat HP 50) se rastvaraju u smeši sledećih rastvarača: aceton 13 L, etanol 94% po težini denaturizovano sa 2% ketona 13.5L i demineralizovana voda 2,5L. 0,240 ricinusovog ulja se dodaje gotovom rastvoru kao sredstvo za omekšavanje i stavlja u porcije na jezgra tableta na uobičajeni način.

Film-obloga: Pošto je sušenje završeno, suspenzija sledećeg sastava se stavlja kao film-obloga u istom aparatu: talk 0,340 kg, titanijum(VI)oksida Cronus RN 56 0,4 kg, obojeni lak L crveni lak 86837 0,324 kg, Eudragit E 12,5 % 4,8 kg i polietilen glikol 6000 pH 11 XI 0,12 kg u smeši rastvarača sledećeg sastava: 2-propanol 8,17 kg,aqua demineralisata0,2 kg i glicerintriacetat (Triacetin) 0,6 kg.

Na želudačnu kiselinu otporne mikro-tablete se onda pune u kapsule od tvrdog želatina neto težine od 500,0 mg i pečate.

Primer 4

Pripremanje enterično- obloženih mikro- tableta u kapsule koje sadrže 120, 0

mg dimetilfumarata, koje odgovaraju 96 mg fumarne kiseline

Preduzimanjem neophodnih mera bezbednosti (maska za disanje, rukavice, zaštitno odelo, itd.), 12 kg dimetilfumarata se drobi i homogenizuje kroz sito 800. Onda se priprema smeša inertnog punioca sledećeg sastava: 17,5 kg derivata škroba (STA-RX 1500®), 0,30 kg mikro-kristalne celuloze (Avicel PH 101®), 0,75 kg PVP (Kollidon® 120), 4 kg Primogel®-a, 0,25 kg koloidne silicijumove kiseline (Aerosil®). Aktivni sastojak se dodaje ukupnoj puder smeši, meša, homogenizuje putem sita 200 i postupa se sa 2% vodenim rastvorom polivinil pirolidona (Kollidon® 25) na uobičajeni način u vezivne granule, i potom meša sa spoljašnom fazom u suvom obliku. Potonje se sastoji od 0,5 kg magnezijum stearata i 1,5 kg talka.

Potom se puder smeđa presuje uobičajenim postupkom u konveksne tablete koji imaju bruto težinu od 10,0 mg i prečnik od 2,0 mm.

Da bi se postigla otpornost na želudačni fluid, rastvor od 2,25 kg hidroksi propil metil celuloza ftalata (HPMCP, Pharmacoat HP® 50) se rastvara u mešti sledećih rastvarača: 13 litra aceton, 13,5 litara etanola (94% po težini denaturizovano sa 2% ketona) i 1,5 Laqua demineralisata.Onda se dodaje ricinusovog ulja (0,24 kg) gotovom rastvoru kao sredstvo za omekšavanje i stavlja se na tabletna jezgra na uobičajeni način.

Posle sušenja suspenzija sledećeg sastava se stavlja u istu aparaturu kao obloga od filma: talk 0,34 kg, titanijum(VI)oksid Cronus RN 56® 0,4 kg,

obojeni lak L-crveno 86837 0,324 kg, Eudragit E 12,5%® 4,8 kg i polietilen glikol 6000 pH 11 XI 0,12 kg u smešu rastvarača sledećeg sastava: 8,17 kg 2-propanol, 0,2 kg demineralizovana voda i 0,6 kg glicerintriacetat (Triacetin®).

Enterično-obložene mikro-tablete se onda pune u kapsule od tvrdog želatina neto težine od 400 mg i potom pečate.

Primer 5

Pripremanje enterično- obloženih mikro tableta u kapsule koje sadrže 120,

mg dimetilfumarata, što odgovara 96 mg fumarne kiseline

12 kg dimetilfumarata se zdrobi i homogenizuje kako je to gore opisano. Onda se priprema smeša inertnog punioca sledećeg sastava: 23,2 kg mikro-kristalna celuloza (Avicel PH 200®), 3 kg kroskarmaloza natrijum (AC-Si-SOL-SD-711), 2,56 kg talk, 0,1 kg anhidrovane silicijumove kiseline (Aerosil® 200) i 1 kg Mg stearata. Potom se puder smeša presuje uobičajenim postupkom u konveksne tablete koje imaju bruto težinu od 10,0 mg i prečnik od 2,0 mm.

Posle toga, priprema se rastvor od 0,94 kg Eudragit® L u izopropanolu koji dodatno sadrži 0,07 kg dibutilftalata. Ovaj rastvor se rasprši na jezgra

tableta. Onda se disperzija od 17,32 kg Eudragit® L D-55 i smeša od 2,8 kg mikro-talka, 2 kg Macrogol 6000 i 0,07 kg dimetikona u vodi priprema i rasprši na jezgra.

Potom, enterično-obložene mikro-tablete se pune u kapsule od tvrdog želatina pri neto težini od 650 mg i pečate.

Primer 6

NF- kappaB translokacija u ćelijsko jezgro

NF-kappaB (p65) se umeće u vektor pEGFP-C1 koji sadrži EGFP (zeleni fluorescentni protein) povezan sa citomegalovirus promoterom (Clontech). Ovo vodi ekspresiji fluorescentnog NF-kappaB. HUVEC ćelije se postave na ploču između trećeg i petog prolaza u želatinom obložene ploče kulture sa 12 bazenčića (Costar) i uzgajaju do 80 ili 90% slivanja, svaka pojedinačno. Onda se ove ćelije podvrgavaju izmenjenoj infekciji upotrebom postupka taloženja kalcijumfosfata. Specifično, ćelije se postavljaju u uslove sa Dulbecco-vim modifikovanim Eagles medijumom (DMEM), taloga koji sadrži 1 jag DNK po bezenčiću dodat posle 24 sata i ćelije se inkubiraju dodatnih četiri sata. Posle pranja sa HBSS (Hanks-ov balansirani rastvor soli), medijum kulture se dodaje i ćelije uzgajaju narednih 18 sati pre nego što se stimulišu.

Radi eksperimenata, ćelije su postavljaju u uslove sa 40 jaM/L

dimetilfumarata, paralelnim pripremanjima bez DNK koja imaju ulogu kontrole. 2 sata posle početka tretiranja pod uslovim ćelije se stimulišu sa 10 ng/mL TNF-a tokom vremena navedenog u tabeli 1.

Posle toga, ćelije se podvrgavaju rastvaranju, površinski sloj se odbacuje a ćelijska jezgra skupljaju u Dounce puferu sa inhibitorom proteaze (10 mM

tris-HCI, pH 7,6, 0,5 mM MgCI, 10 jag/mL leupeptin, 10ng/mL aprotinin, 1

mM fenil metil sulfonil fluorid, 1,8 mg/mL jodoacetamida). Posle 10 minuta centrifugiranja na 1200 g, 4°C, ćelijska jezgra se analiziraju na FAC scanflovv citometru (Becton Dickinson).

Primer 7

Inhibicija sa NF- kappaB stimulisanog zapisa

Trostruko ponavljanje AP-1 položaja saglasnosti (položaj vezivanja) (48bp, 3

x TGTGA- TGACTCAGGTT) i trostruko ponavljanje NF-kapaB položaja saglasnosti (60bp, 3 x AATCGT GGAATTTCCTCTGA), obiđeni sa boka od strane Spel položaja vezivanja (nije prikazano) se umeću u Spel položaj pTK-UBT-luc vektora (de Martin, Gene 124, 137-138, 1993). 1,3 kb konstrukt E-selektin promotera koji se prostire od bp-1285 do bp+482 se umeće u Ndel položaj pMAM Neo-luc vektora (Clontech).

HUVEC ćelije se podvrgavaju tranfekciji sa tako dobijenim konstruktima kako je to opisano u primeru 6. Za navedenu transfekciju, dodaje se 2,5 [ig značajnog promoter konstrukta po bazenčiću. Da bi se verifikovala efikasnost transfekcije, ko-transfekcije se obavljaju sa 500 ng pSV-beta galaktozidaza kontrolnog vektora (Promega Corp., Madison, Wl, U.S.A.) kao kontrola u svakom eksperimentu. Dva dana posle transfekcije ćelije se stimulišu tokom 2 sata sa 10 ng/mL TNF-alfa sa i bez dodavanja 6 (.ig/mL

dimetilfumarata (DMF). Ćelije se onda oberu tripsinacijom, granuliraju,

operu i re-suspenduju u 200 jaL "puferu za rastvaranje reportera" (Promega)

tokom 15 minuta, kako je to propisano od strane proizvođača.

Aktivnost luciferaze se meri sredstvima Berthold AuotoLumat LB9507 luminometra upotrebom luciferaza test sistema (Promega). Aktivnost beta-galaktozidaze se utvrđuje upotrebom Promega beta-galaktozidaza enzim test sistema. Aktivnosti luciferaze dobijene sa značajnim promoter konstruktima se normalizuju prema beta-galaktozicaza aktivnosti. Varirajuća širina aktivnosti beta-galaktozidaze u okviru pojedinačnih eksperimenata je ispod 10%. Tabela 2 pokazuje pojedinačne rezultate x-putavis- a- visosnovne linije.

Tabela 2 pokazuje da dimetilfumarat inhibira sa TNF izazvanu transkripciju od NF-kappaB zavisnog gena, ali ne transkripciju AP-1 zavisnog gena. Prema tome inhibicija dimetilfumaratom je specifična za NF-kappaB.

Claims (18)

1. Upotreba jednog ili više derivata fumarne kiseline za pripremanje farmaceutskog sastava za lečenje bolesti na koje se može uticati sa NF-kappaB.

2. Upotreba prema Zahtevu 1, karakterisana time da je derivat fumarne kiseline odabran od grupe koja se sastoji od dialkil estara fumarne kiseline i monoalkil estara fumarne kiseline, koji mogu opciono biti supstituisani, u obliku slobodne kiseline ili njihovih soli i od njih spravljenih smeša.

3. Upotreba prema Zahtevu 2, pri čemu dialkil estar fumarne kiseline odgovara formuli pri čemu F^i R2koji mogu biti isti ili različiti, nezavisno predstavljaju linearni, račvasti, ciklični, zasićeni ili nezasićeni C^ 24 alkil radikal ili C5_2oaril redikal i ovi radikali su opciono supstituisani sa halogenom (F, Cl, Br, J), hidroksi, C1.4alkoksi, nitro ili cijano.

4. Upotreba prema jednom od Zahteva 2 i 3, karakteristična po tome što su radikali R1i R2metil, etil, n-propil, izopropil, n-butil, sek-butil, t-butil, pentil, ciklopentil, 2-etilheksil, heksil, cikloheksil, heptil, cikloheptil, oktil, vinil, alil, 2-hidroksietil, 2- ili 3-hodroksipropil, 2,3-dihidroksipropil, 2-metoksietil, metoksimetil ili 2- ili 3-metoksipropil.

5. Upotreba prema Zahtevu 2 pri čemu monoalkil estar fumarne kiseline odgovara formuli pri čemu - Rtje kako je to definisano u Zahtevima 3 i 4 - A je vodonik, alkali i alkalni katjon zemnih metala ili fiziološki kompatibilni katjon prelaznog metala, pretpostavljeno odabran od Li<+>, Na<+>, K<+>, Mg2\Ca<2>+,Zn<2+>, Fe2+ i Mn<2+>, i - n je 1 ili 2 i odgovara valenci A.

6. Upotreba prema bilo kom od prethodnih zahteva, karakterisana time da je derivat fumarne kiseline jedan ili više derivata odabranih od grupe koja obuhvata dimetil estar fumarne kiseline, dietil estar fumarne kiseline, metiletil estar fumarne kiseline, metilfumarna kiselina, etilfumarna kiselina, kalcijummetilfumarat, kalcijumetilfumarat, magnezijummetilfumarat, magnezijumetilfumarat, cinkmetilfumarat, cinketilfumarat, gvožđemetilfumarat i gvožđeetilfumarat i smeše nastale od njih.

7. Upotreba prema Zahtevu 6, karakterisana time da je derivat fumarne kiseline dimetil estar fumarne kiseline (dimetilfumarat).

8. Upotreba jednog ili više derivata fumarne kiseline za pripremanje farmaceutskog sastava za lečenje bolesti na koje se može uticati sa NP-kappaB, odabranih iz grupe bolesti u koje spadaju: progresivna sistemska sklerodermija, osteochondritis svphilitica (Wegener-ova bolest), cutis marmorata (livedo reticularis), Behcet-ova bolest, panarteriitis, ulcerozni kolitis, vaskulitis, osteoartritis, podagra, aretrioskleroza, Reiter-ova bolest, pulmonalna granulomatoza, tipovi encefalitisa, endotoksični šok (septični/toksični šok), sepsa, pneumonija, encefalomijelitis, anorexia nervosa, hepatitis (akutni hepatitis, hronični hepatitis, toksični hepatitis, alkoholom izazvani hepatitis, virusni hepatitis, žutica, insuficijencija jetre i citomegalovirusni hepatitis), Rennert T-limfomatoza, mesangial nefritis, post-angioplastična restenoza, reperfuzioni sindrom, citomegavirusna retinopatija, adenovirusne bolesti kao što su adenovirusni nazebi, adenovirusna faringokonjuktivistična groznica i adenovirusna oftalmija, AIDS, Guillain-Barre sindrom, post-herpetična ili post-zoster neuralgia, inflamatorna demijelizirajuća polineuropatija, mononeuropatija multipleks, muskoviscidoza, Bechterew-ova bolest, Barett-ov ezofagus, EBV (Epstein-Barr virusna) infekcija, srčano remodelovanje, intersticijalni cistitis, dijabetis mellitus tip II, humani radioaktivno senzibilni tumor, multi-otpornost malignih ćelija na hemoterapeutska sredstva (otpornost mulitleka u hemoterapiji), granualoma annulare i kanceri kao što su karcinom dojke, karcinom debelog creva, melanom, primami karcinom ćelije jetre, adenokarcinom, Kapošijev sarkom, karcinom prostate, leukemija kao što je akutna mijeloidna leukemija, višestruki mijelom (plasmocvtoma), Burkitt-ov limfosarkom i Castelman-ov tumor.

9. Upotreba prema Zahtevu 8, karakterisana time što je derivat fumarne kiseline odabran od grupe koaj se sastoji od dialkil estara fumarne kiseline i monoalkil estara fumarne kiseline u obliku slobodne kiseline ili soli ili od njih spravljenih smeša.

10. Upotreba prema Zahtevu 9, pri čemu dialkil estar fumarne kiseline odgovara formuli pri čemu Rii R2koji mogu biti isti ili različiti, nezavisno predstavljaju linearni, račvasti, ciklični, zasićeni ili nezasićeni C-i_24alkil radikal ili C5.20aril redikal i ovi radikali su opciono supstituisani sa halogenom (F, Cl, Br, J), hidroksi, d.4 alkoksi, nitro ili cijano.

11. Upotreba prema jednom od Zahteva 9 i 10, karakteristična po tome što su radikali Rii R2metil, etil, n-propil, izopropil, n-butil, sek-butil, t-butil, pentil, ciklopentil, 2-etilheksil, heksil, cikloheksil, heptil, cikloheptil, oktil, vinil, alil, 2-hidroksietil, 2- ili 3-hodroksipropil, 2,3-dihidroksipropil, 2-metoksietil, metoksimetil ili 2- ili 3-metoksipropil.

12. Upotreba prema Zahtevu 9 pri čemu monoalkil estar fumarne kiseline odgovara formuli pri čemu - Rije kako je to definisano u Zahtevima 3 i 4 - A je vodonik, alkali ili alkalni katjon zemnog metala ili fiziološki kompatibilni katjon prelaznog metala, pretpostavljeno odabran od Li<+>, Na<+>, K<+>, Mg2+, Ca2+, Zn2+, Fe2+ i Mn2+, i - n je 1 ili 2 i odgovara valenci A.

13. Upotreba prema bilo kom od Zahteva 8 do 12, karakterisana time što jedna jedinica doze farmaceutskog sastava sadrži količinu derivata fumarne kiseline koja odgovara 1 do 500 mg, pretpostavljeno 10 do 300 mg a najpoželjnije 10 do 200 mg fumarne kiseline.

14. Upotreba prema bilo kom od ranijih Zahteva 8 do 13 za pripremanje farmaceutskog sastava za oralnu, parenteralnu, rektalnu, transdermalnu, dermalnu, nazalnu, pulmonalnu (inhalacija) ili oftalnu administraciju, pretpostavljeno sa oralnu administraciju.

15. Upotreba prema Zahtevu 14 gde je farmaceutski sastav za oralnu administraciju prisutan u obliku tableta jedinične doze kao tableta, mikro-tableta, mikro-pilula ili granulata (navedene mikro-tablete, mikro-pilule ili granulat su opciono kapsulirane ili napunjene u kesice), kapsula ili rastvora koji se piju.

16. Upotreba prema zahtevu 15, karakteristična po tome što su čvrsti oblici doza obezbeđeni sa enteričnim oblaganjem.

17. Upotreba prema Zahtevu 8, karakteristična po tome što oblici doze farmaceutskog sastava pretpostavljeno sadrže bilo individualno ili u primesi: 10 do 500 mg dialkilfumarata, posebno dimetilfumarata i/ili dietilfumarata; 10 do 500 mg kalcijumalkilfumarata, posebno kalcijummetilfumarata i/ili kalcijumetilfumarata 0 do 250 mg cinkalkilfumarata, posebno cinkmetilfuamarata i/ili cinketilfumarata, 0 do 250 mg alkilvodonikfumarata, posebno metilvodonikfumarata i/ili etilvodonikfumarata; i 0 do 250 mg magnezijumalkilfumarata, posebno magnezijummetilfumarata i/ili magnezijumetilfumarata ukupno gore pomenutih količina koje odgovaraju ekvivalentu od 10 do 500 mg, pretpostavljeno 10 do 300 mg a najpoželjnije 100 mg fumarne kiseline.

18. Upotreba prema jednom od Zahteva 15 ili 16, karakteristična po tome što je sastav prisutan u obliku mikro-tableta ili mikro-pilula koje imaju veličinu od <5000 |am, pretpostavljeno veličine od 300 do 1000 |am za pilule i 1000 do 2500 (im za mikro-tablete. Fumaric Acid Derivativesa$. NF-kappaB Inhibitor The present invention relates to the use of one or more fumaric acid derivatives as NF-kappaB inhibitor. At the same time, the present invention relates to the use of the fumaric acid derivatives for preparing a pharmaceutical composition for treating diseases that may be influenced by NF-kappaB. It is known that pharmaceutical preparations such as fumaric acid which, upon biological degradation after administration, enter into the citric acid cycle or are part thereof gain increasing therapeutic significance - especially when given in high dosages - since they can alleviate or heal diseases caused cryptogenetically. In addition, fumaric acid inhibits the growth of the Ehrlich ascites tumour in mice, reduces the toxic effects of mitomycin C and aflatoxin and displays anti-psoriatic and anti-microbial activity. The most important practical application is the treatment of psoriasis with various fumaric acid derivatives which has already been described in a number of patents, for example EP 0 188 479, DE 25 30 372, DE 26 21 214 or EP 0 312 697. Another use of certain fumaric acid derivatives, namely of the alkyl hydrogen fumarates, is disclosed in DE 197 21 099.6 and DE 198 53 487.6 according to which these specific fumaric acid derivatives are described for treating auto-immune diseases such as polyarthritis, multiple sclerosis and graft-versus-host reaction. In addition, DE 198 53 487.6 and DE 198 39 566.3 teach the use of alkyl hydrogen fumarates and dialkyl fumarates in transplantation medicine. Even though individual investigations of the action mechanism of fumaric acid derivatives in the treatment of psoriasis have been carried out, no specific information exists on this topic. The NF-kappaB (nuclear factor kappaB) is a transcription factor of eukaryotic cells. NF-kappaB belongs to the family of Rel proteins, a class of transcription factorscharacterised bya so-called Rel domain. The Rel domain has been named after the first member found in an avian virus as an oncogen. Specific sites of this homologous Rel domain (Rel homology domain = RHD) which consists of 300 amino acids are responsible for the DNA bonding to the kappaB sites, the dimeri-sation with other proteins of the Rel family and the interaction with I-kappaB. So far. five members of the Rel family are known in mammals. These are c-Rel. NF-kappaB 1 (pl05/p50), NF-kappaB2 (pl00/p52) and RelB. In theory, these five members of the Rel protein family may combine into any form of homo- and heterodimers, even though only a few specific combinations have been observedin vivo.The classic and bestcharacterisedNF-kappaB molecule is a heterodimer of the p50/p65 sub-units NF-kappaB 1/RelA. This heterodimer is the most common complex and is found in practically all cell types. After the cellular activation and the dissociation of I-kappaB, the NF-kappaB heterodimer p50/p65 migrates into the cell nucleus where it binds to the consensus sequence 5'-GGGRNNYYCC-3'. In this process, the p50 sub-unit primarily serves as the DNA-binding sub-unit, while the p65 sub-unit provides the transactivation function. As a result of the different combinations, each of these heterodimers displays unique characteristics as far as cell type specificity, preferences with regard to DNA-bonding, differential interaction with I-kappaB isoforms, differential activation requirements and the kinetics of activation are concerned. The rapid inducibility of NF-kappaB is attributed to the fact that the factor is present in the cytoplasm in an inactive form, namely in a complex bonded to the NF-kappaB inhibitor I-kappaB. Therefore, no new protein synthesis is required for activation, but merely the solution of this complex with I-kappaB or degradation of this inhibitor and subsequent translocation of the now active NF-kappaB dimer into the nucleus. NF-kappaB may be activated by a large variety of physiological and non-physiological stimuli. These include cytokines, mitogenes, viruses, viral products, the cross-linking of antigen receptors on T- and B-lymphocytes, calcium ionophores, phorbol esters, UV-rays, oxidation stress, phosphatase inhibitors and others. The range of the many NF-kappaB regulated or activated genes is just as broad, the transcription of which is activated, induced or enhanced by the bonding of the heterodimer to the consensus sequence as described above. Especially TNF-alpha. IL-1, IL-2 and lipopolysaccharides may be mentioned as important stimulants. The regulated genes generally comprise genes involved in the immune function, inflammation response, cell adhesion, cell growth, but also cell death. Genes of cell adhesion molecules, cytokines, cytokine receptors, acute phase proteins, growth factors and viral genes should be mentioned in particular. Special among the genes induced by NF-kappaB are the genes for interferon-B, for the light chain of the immunoglobulin, for the T-cell receptor, for TNF-a and TNF-B and for the tissue factor (CD 142), formerly called tissue tromboplastin or factor III. Owing to its central role in the regulation of immune reactions and inflammation responses shown above and its involvement in the regulation of tissue factors, cytokines etc. it was assumed that advantages similar to those already known from anti-inflammatory agents may be expected from the development of selective inhibitors for the transcription factor NF-kappaB. Steroidal anti-inflammatory agents, interferons or cyclosporine may be named as examples. Surprisingly, it has now been found that individual fumaric acid derivatives or mixtures thereof have an NF-kappaB inhibiting effect. This effect may preferably be used for the preparation of a pharmaceutical composition containing these fumaric acid derivatives individually or in admixture for the therapy of diseases that are mediated or may be influenced by NF-kappaB. In particular, diseases that may be influenced by NF-kappaB are progressive systemic sclerodermia, osteochondritis syphilitica (Wegener's disease), cutis marmorata{ livedo reticularis),Behcet disease, panarteritis, colitis ulcerosa, vasculitis, osteoarthritis, gout, arteriosclerosis, Reiter's disease, pulmonary granulomatosis, types of encephalitis, endotoxic shock (septic-toxic shock), sepsis, pneumonia, encephalomyelitis, anorexia nervosa, hepatitis (acute hepatitis, chronic hepatitis, toxic hepatitis, alcohol-induced hepatitis, viral hepatitis, jaundice, liver insufficiency and cytomegaloviral hepatitis), Rennert T-lymphomatosis, mesangial nephritis, post-angioplastic restenosis, reperfusion syndrome, cytomegaloviral retinopathy, adenoviral diseases such as adenoviral colds, adenoviral pharyngoconjunctival fever and adenoviral ophthalmia, AIDS, Guillain-Barre syndrome, post-herpetic or post-zoster neuralgia, inflammatory demyelinising polyneuropathy, mononeuropathia multiplex, mucoviscidosis, Bechterew's disease, Barett oesophagus, EBV (Epstein-Barr virus) infec-tion, cardiac remodeling, interstitial cystitis, diabetes mellitus type II, human tumour radiosensitisation, multi-resistance of malignant cells to chemotherapeutic agents (multidrug resistance in chemotherapy), granuloma annulare and cancers such as mamma carcinoma, colon carcinoma, melanoma, primary liver cell carcinoma, adenocarcinoma, kaposi's sarcoma, prostate carcinoma, leukaemia such as acute myeloid leukaemia, multiple myeloma (plasmocytoma), Burkitt lymphoma and Castleman rumour. According to the invention, one or more fumaric acid derivatives selected from the group consisting of fumaric acid dialkyi esters and fumaric acid monoalkyl esters in the form of the free acid or in the form of salts and mixtures thereof are preferably used for NF-kappaB inhibition and for preparing the pharmaceutical composition. The fumaric acid dialkyi esters preferably correspond to the formula wherein R]andRi,which may be the same or different, independently represent a linear, branched, cyclic, saturated or unsaturated Q_24alkyl radical or a C5.20aryl radical and these radicals are optionally substituted with halogen (F, CI, Br, I), hydroxy, Cm alkoxy, nitro or cyano. The radicals R]and R2are preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, t-butyl, pentyl, cyclopentyl, 2-ethylhexyl, hexyl, cyclohexyl, heptyl, cycloheptyl, octyl, vinyl, allyl, 2-hydroxyethyl, 2--or 3-hydroxypropyl, 2,3-dihydroxypropyl, 2-methoxyethyl, methoxymethyl or 2- or 3-methoxypropyl. The fumaric acid monoalkyl esters preferably correspond to the formula wherein R]is as defined above, A is hydrogen, an alkali or alkaline earth metal cation or a physiologically compatible transition metal cation, preferably selected from Li , Na', K , Mg'', Ca~ , Zn~ , Fe~' and Mn" , and n is 1 or 2 and corresponds to the valence of A The invention preferably uses one or more fumaric acid derivative(s) selected from the group comprising fumaric acid dimethyl ester, fumaric acid diethyl ester, fumaric acid methylethyl ester, methyl hydrogen fumarate, ethyl hydrogen fumarate, magnesium methyl fumarate, magnesium ethyl fumarate, zinc methyl fumarate, zinc ethyl fumarate, iron methyl fumarate, iron ethyl fumarate, calcium methyl fumarate and/or calcium ethyl fumarate,. According to the invention, the fumaric acid derivatives for preparing the pharmaceutical composition are preferably used in such an amount that one dosage unit of said pharmaceutical composition contains an amount of fumaric acid derivative^) corresponding or equivalent to 1 to 500 mg, preferably 10 to 300 mg and most preferably 10 to 200 mg of fumaric acid. Preferred forms of administration for the pharmaceutical composition are oral, parenteral, rectal transdermal, dermal, nasal, pulmonal (inhalation) or ophthal administration (in the form of eye drops), oral administration being preferred. The composition will then be present in a suitable form for each type of administration. When administered orally, the pharmaceutical composition is present in the form of single unit dose tablets, micro-tablets (multiple unit dose tablets) or mini-tablets, micro-pellets or granulate (said micro-tablets, micro-pellets or the granulate optionally being encapsulated or filled into sachets), capsules or solutions for drinking.Ina preferred embodiment, solid dosage or administration forms are provided with an enteric coating. Such a coating may also be provided on encapsulated or filled dosage forms. In case of parenteral administration by injection (i.v., i.m., s.c, i.p.), the composition is present in a suitable form. All customary liquid carriers suitable for injec-tions may be used. The pharmaceutical composition may preferably contain either individually or in admixture: 10 to 500 mg of dialkyi fumarate. especially dimethyl fumarate and/or diethyl fumarate; 10 to 500 mg of calcium alkyl fumarate. especially calcium methyl fumarate and/or calcium ethyl fumarate; 0 to 250 mg of zinc alkyl fumarate. especially zinc methyl fumarate and/or zinc ethyl fumarate; 0 to 250 mg of alkyl hydrogen fumarate. especially methyl hydrogen fumarate and/or ethyl hydrogen fumarate; and 0 to 250 mg magnesium alkyl fumarate, especially magnesium methyl fumarate and/or magnesium ethyl fumarate, the total of the above-mentioned amounts corresponding to an equivalent of 10 to 500 mg, preferably 10 to 300 mg and most preferably 100 mg of fumaric acid. Preferred compositions of the invention contain only dimethyl fumarate in an amount of 10 to 300 mg. According to a particularly preferred embodiment, the composition is present in the form of micro-tablets or micro-pellets. These preferably have a size or mean diameter of < 5000 urn, more preferably 300 to 2500 urn, especially 300 to 1000 um for pellets and 1000 to 2500 um for the micro-tablets. By administering the fumaric acid derivatives in the form of micro-tablets, which is preferred in accor-dance with the invention, gastro-intestinal irritations or side effects which cannot be ruled out in case of administration of customary single-unit dose tablets may be further reduced. This is probably due to the fact that micro-tablets, preferably micro-tablets with enteric coating, are already dispersed in the stomach and therefore reach the intestinal tract in portions where the active ingredients are released in locally smaller doses while the overall dose remains the same. This, in turn, helps avoid local irritation of the epithelial intestinal cells resulting in improved gastro-intestinal tolerance of the micro-tablets in comparison with conventional tablets. For example, the fumaric acid derivatives contained in the compositions of the invention are prepared by the process described in EP 0 312 679. Production examples In principle, the oral compositions of the invention in the form of tablets or micro-tablets may be prepared by classic tabletting procedures. Instead of such classic tabletting procedures other methods for preparing tablets may be used, such as direct tabletting as well as processes for preparing solid dispersions according to the melt process or the spray drying process. The tablets may be provided with an enteric coating. The enteric coating may be applied in a classic coating pan or sprayed on. The coating may also be applied with a Boegel coating apparatus. In addition, the tablet may be provided with a film coat. In order to explain the use according to the invention, various examples for preparing preferred drugs are given below. These examples are intended to illustrate, but not to limit the invention. Example 1 Preparation of film tablets with an enteric coating containing 100. 0 mg of monomethyl fumarate- Ca salt, which corresponds to 78 mg of fumaric acid Taking the necessary precautions (breathing mask, gloves, protective clothing, etc.), 10 kg of monomethyl fumarate-Ca salt are crushed, mixed intensely and homogenised by means of a sieve 800. Then an excipient mixture of the following composition is prepared: 21 kg of starch derivative (STA-RX 1500®), 2 kg of micro-crystalline cellulose (Avicel PH 101®), 0.6 kg of polyvinyl pyrrolidone (PVP, Kollidon® 25), 4 kg of Pnmogel®, 0.3 kg of colloidal silicic acid (Aerosil®). The active ingredient is added to the entire powder mixture, mixed, homogenised by means of a sieve 200 and processed with a 2 % aqueous solution of polyvinyl pyrrolidone (PVP, Kollidon® 25) in the usual manner into binder granules, and then mixed with the outer phase in a dry state. The latter consists of 2 kg of a so-called FST complex containing 80 % of talcum, 10 % of silicic acid and 10 % of magnesium stearate. Thereafter the mixture is pressed into convex tablets with a weight of 400 mg and a diameter of 10.0 mm by the usual method. Instead of these classic compaction methods, other methods such as direct compaction or solid dispersions according to the melting and spray drying method may also be used for preparing tablets. Enteric coating: A solution of 2.250 kg of hydroxy propyl methyl cellulose phthalate (HPMCP, Pharmacoat HP® 50) is dissolved in a solvent mixture consisting of 2.50 litres of demineralised water. 13 litres of acetone Ph. Helv. VII and 13 litres of ethanol (94 % by weight) and then 0.240 kg of castor oil (Ph. Eur. II) is added to the solution. The solution is poured or sprayed in portions onto the tablet cores in a coating pan in a conventional manner or applied by means of a fluidised-bed apparatus of the appropriate structure. After drying, the film coating is applied. Said coating consists of a solution of Eudragit E 12.5 %® 4.8 kg, talcum Ph. Eur. II 0.34 kg, titanium(VI) oxide Cronus RN 56® 0.52 kg, coloured lacquer ZLT-2 blue (Siegle) 0.21 kg, and polyethylene glycol 6000 Ph. Helv. VII 0.12 kg in a solvent mixture of 8.2 kg of 2-propanol Ph. Helv. VII, 0.06 kg of glycerine triacetate (Triacetin®) and 0.2 kg of demineralised water. After homogenous distribution in the coating pan or the fluidised bed, the mixture is dried and polished in the usual manner. Example 2 Preparation of enteric coated capsules containing 86. 5 mg of monoethvl fumarate-Ca salt and 110. 0 mg of dimethyl fumarate, which corresponds to a total of 150 mg of fumaric acid Taking the necessary precautions (breathing mask, gloves, protective clothing, etc.), 8.65 kg of monoethyl fumarate-Ca salt and 11 kg of dimethyl fumarate are intensely mixed with a mixture consisting of 15 kg of starch, 6 kg of lactose Ph. Helv. VII, 2 kg of micro-crystalline cellulose (Avicel®), 1 kg of polyvinyl pyrrolidone (Kollidon® 25) and 4 kg of Primogel® and homogenised by means of a sieve 800. Together with a 2 % aqueous solution of polyvinyl pyrrolidone (Kollidon® 25) the entire powder mixture is processed in the usual manner into a binder granulate and mixed with the outer phase in the dried state. Said outer phase consists of 0.35 kg of colloidal silicic acid (Aerosil®), 0.5 kg of Mg stearate and 1.5 kg of talcum Ph. Helv. VII. The homogeneous mixture is then filled in portions of 500.0 mg into appropriate capsules which are then provided with an enteric (gastric-acid resistant) coating consisting of hydroxy propyl ethyl cellulose stearate and castor oil as softening agent by a known method. Instead of hard gelatine capsules, the mixture may also be filled into appropriate gastric acid-resistant capsules. w;hich consist of a mixture of cellulose acetate phthalate (CAP) and hydroxy propyl ethyl cellulose phthalate (HPMCP). Example 3 Preparation of enteric- coated micro- tablets in capsules containing 87. 0 mg of monoeth<y>l fumarate- Ca salt. 120 mg of dimethyl fumarate. 5. 0 mg of monoethvl fumarate- Mg salt and 3. 0 mg of monoethvl fumarate- Zn salt, which corresponds to a total of 164 mg of fumaric acid(" forte "tablets) Taking the necessary precautions (breathing mask, gloves, protective clothing, etc.). 8.7 kg of monoethyl fumarate-Ca salt, 12 kg of dimethyl fumarate, 0.5 kg of monoethyl fumarate-Mg salt and 0.3 kg of monoethyl fumarate-Zn salt are crushed, intensely mixed and homogenised by means of an sieve 800. Then an excipient mixture of the following composition is prepared: 18 kg of starch derivative (STA-RX 1500), 0.3 kg of micro-crystalline cellulose (Avicel PH 101),

0.75 kg of PVP (Kollidon 120), 4 kg of Pnmogel, 0.25 kg of colloidal silicic acid (Aerosil). The entire powder mixture is added to the active ingredient mixture, homogenised by means of a 200 sieve, processed m the usual manner with a 2 % aqueous solution of polyvinyl pyrrolidone (Kollidon K25) to obtain a binder granulate and mixed in a dry state with the outer phase consisting of 0.5 kg of magnesium stearate and 1.5 kg of talcum. Then the powder mixture is pressed by the conventional method into convex micro-tablets with a gross mass of 10.0 mg and a diameter of 2.0 mm. Instead of this classic tabletting method other methods for making tablets such as direct tabletting or solid dispersions by the melt method and the spray drying method may also be used. The gastric acid-resistant coating may be poured or sprayed on in a classic coating pan or applied in a fluidised-bed apparatus. In order to achieve resistance to gastric acid, portions of a solution of 2.250 kg of hydroxy propyl methyl cellulose phthalate (HPMCP, Pharmacoat HP 50) are dissolved in a mixture of the following solvents: acetone 13 1, ethanol 94 % by weight denatured with 2 % ketone 13.5 1 and demineralised water 2.5 1. 0.240 kg of castor oil are added as softening agent to the finished solution and applied in portions to the tablet cores in the usual manner. Film-coat: After drying is completed, a suspension of the following composition is applied as a film-coat in the same apparatus: talcum 0.340 kg. titanium(VI) oxide Cronus RN 56 0.4 kg, coloured lacquer L red lacquer 86837 0.324 kg, Eudragit E 12.5 % 4.8 kg and polyethylene glycol 6000 pH 11 XI 0.12 kg in a solvent mixture of the following composition: 2-propanol 8.17 kg,aqua demineralisata0.2 kg and glycerine triacetate (Triacetin) 0.6 kg. The gastric acid-resistant micro-tablets are then filled into hard gelatine capsules at a net weight of 500.0 mg and sealed. Example 4 Preparation of entenc- coated micro- tablets in capsules containing 120. 0 mg of dimethyl fumarate, which corresponds to 96 mg of fumaric acid Taking the necessary precautions (breathing mask, gloves, protective clothing, etc.), 12 kg of dimethyl fumarate are crushed and homogenised by means of a sieve 800. Then an excipient mixture of the following composition is prepared:

17.5 kg of starch derivative (STA-RX 1500®), 0.30 kg of micro-crystalline cellulose (Avicel PH 101®), 0.75 kg of PVP (Kollidon® 120), 4 kg of Primogel®, 0.25 kg of colloidal silicic acid (Aerosil®). The active ingredient is added to the entire powder mixture, mixed, homogenised by means of a sieve 200 and processed with a 2 % aqueous solution of polyvinyl pyrrolidone (Kollidon® 25) in the usual manner into binder granules, and then mixed with the outer phase in a dry state. The latter consists of 0.5 kg of magnesium stearate and 1.5 kg of talcum. Thereafter the powder mixture is pressed into convex tablets having a gross weight of 10.0 mm and a diameter of 2.0 mm by the usual method. To achieve resistance against gastric fluid, a solution of 2.25 kg of hydroxy propyl methyl cellulose phthalate (HPMCP, Pharmacoat HP® 50) is dissolved in a mixture of the following solvents: 13 litres of acetone, 13.5 litres of ethanol (94 % by weight denatured with 2 % of ketone) and 1.5 1of aqua demineralisata.Then castor oil (0.24 kg) is added to the finished solution as a softening agent and applied onto the tablet cores in the usual manner. After drying a suspension of the following composition is applied in the same apparatus as a film coat: talcum 0.34 kg, titanium(VI) oxide Cronus RN 56® 0.4 kg, coloured lacquer L-red 86837 0.324 kg, Eudragit E 12.5 %® 4.8 kg and polyethylene glycol 6000 pH 11 XI 0.12 kg in a solvent mixture of the following composition: 8.17 kg of 2-propanol, 0.2 kg of demineralised water and 0.6 kg of glycerine triacetate (Triacetin®). The enteric-coated micro-tablets are then filled into hard gelatine capsules at a net weight of 400 mg and sealed. Example 5 Preparation of enteric- coated micro- tablets in capsules containing 120. 0 mg of dimethyl fumarate. which corresponds to 96 mg of fumaric acid

12 kg of dimethyl fumarate are crushed and homogenised as described above, Then an excipient mixture of the following composition is prepared: 23.2 kg of micro-crystalline cellulose (Avicel PH 200®), 3 kg of croscarmelose sodium (AC-Si-SOL-SD-711), 2. 5 kg of talcum, 0.1 kg of anhydrous silicic acid (Aerosil® 200) and 1 kg of Mg stearate. Thereafter the powder mixture is pressed into convex tablets having a gross weight of 10.0 mm and a diameter of 2.0 mm by the usual method. After that, a solution of 0.94 kg Eudragit® L in isopropanol is prepared which additionally contains 0.07 kg of dibutyl phthalate. This solution is sprayed onto the tablet cores. Then a dispersion of 17.32 kg of Eudragit® L D-55 and a mixture of 2.8 kg of micro-talcum, 2 kg of Macrogol 6000 and 0.07 kg of dimeticon in water is prepared and sprayed onto the cores. Thereafter, the enteric-coated micro-tablets are filled into hard gelatine capsules at a net weight of 650 mg and sealed. Example 6 NF- kappaB translocation into the cell nucleus NF-kappaB (p65) was inserted into the vector pEGFP-Cl which contained EGFP (green fluorescent protein) linked with a cytomegalovirus promoter (Clontech). This leads to the expression of a fluorescent NF-kappaB. HUVEC cells were plated between the third and the fifth passage in gelatine-coated culture plates having 12 wells (Costar) and grown to 80 or 90 % confluence, respectively. Then these cells were subjected to transfection using the calcium phosphate precipita-tion method. Specifically, the cells were conditioned with Dulbecco's modified Eagles medium (DMEM), the precipitate containing 1 ug of DNA per well added after 24 hours and the cells incubated a further four hours. After washing with HBSS (Hanks balanced salt solution), culture medium was added and the cells grown for a further 18 hours before they were stimulated. For the experiments, the cells were conditioned with 40 uM/1 of dimethyl fumarate. parallel preparations without DNA acting as control. 2 hours after com-mencement of conditioning the cells were stimulated with 10 ng/ml TNF-a for the time stated in table 1. After that, the cells were subjected to lysis, the supernatant discarded and the cell nuclei collected in Dounce buffer with protease inhibitor (10 mM tris-HCl, pH 7.6, 0.5 mM MgCl, 10 ug/ml leupeptin, 10 ug/ml aprotinin, 1 mM phenyl methyl sulfonyl fluoride, 1.8 mg/ml iodoacetamide). After 10 minutes of centrifugation at 1200 g, 4°C, the cell nuclei were analysed on an FACscanflow cytometer (Becton Dickinson). This table shows that dimethyl fumarate at a concentration of 40 uM/1 inhibited the TNF-induced translocation of NF-kappaB into the cell nucleus. Example 7 Inhibition of the NF- kappaB stimulated transcription A triple repeat of the AP-1 consensus site (bonding site) (48bp, 3 x TGTGA-TGACTCAGGTT) and a triple repeat of the NF-kappaB consensus site (60 bp, 3 x AATCGT GGAATTTCCTCTGA). flanked by Spel bonding sites (not shown) were inserted into the Spel site of the pTK-UBT-luc vector (de Martin, Gene 124, 137 - 138, 1993). A 1.3 kb construct of the E-selectin promoter extending from bp -1285 to bp +482 was inserted into the Ndel site of the pMAM Neo-luc vector (Clontech). HUVEC cells were subjected to transfection with the constructs thus obtained as described in example 6. For said transfection, 2.5 ug of the pertinent promoter construct per well were added. In order to verify the transfection efficiency, co-transfections were carried out with 500 ng of a pSV-beta galactosidase control vector (Promega Corp., Madison, WI, U.S.A.) as control in each experiment. 2 days after transfection the cells were stimulated for 2 hours with 10 ng/ml TNF-alpha with and without addition of 6 ug/ml dimethyl fumarate (DMF). The cells were then harvested by trypsmation. pelletised, washed and re-suspended in 200 pi of "reporter lysis buffer" (Promega) for 15 min. as prescribed by the manufac-turer. The luciferase activity was measured by means of a Berthold AutoLumat LB9507 luminometer using the luciferase test system (Promega). The beta-galactosidase activity was determined using the Promega beta-galactosidase enzyme test system. The luciferase activities obtained with the pertinent promoter constructs were normalised to the beta-galactosidase activity. The variation width of the beta-galactosidase activity within the individual experiments was below 10 %. Table 2 shows the individual results x-fold vis-a-vis the base line. Table 2 shows that dimethyl fumarate inhibited the TNF induced transcription of a NF-kappaB dependent gene, but not the transcription of an AP-1 dependent gene. Therefore the dimethyl fumarate inhibition is NF-kappaB-specific.

1. The use of one or more fumaric acid derivatives for preparing a pharmaceutical composition for the treatment of diseases that may be influenced by NF-kappaB.

2. The use according to claim 1,characterised in thatthe fumaric acid derivative is selected from the group consisting of fumaric acid dialkyi esters and fumaric acid monoalkyl esters, which may optionally be substituted, in the form of the free acid or its salts and mixtures thereof.

3. The use according to claim 2, wherein the fumaric acid dialkyi ester corre sponds to the formula wherein Rx and R2, which may be the same or different, independently represent a linear, branched, cyclic, saturated or unsaturatedC1- 2 4alkyl radical or a C5-20aryl radical and these radicals are optionally substituted with halogen (F, CI, Br, I), hydroxy, C^ alkoxy, nitro or cyano.

4. The use according to one of the claims 2 and 3,characterised in thatthe radicals Ri and R2are methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, t-butyl, pentyl, cyclopentyl, 2-ethylhexyl, hexyl, cyclohexyl, heptyl, cycloheptyl, octyl, vinyl, allyl, 2-hydroxyethyl, 2- or 3-hydroxypropyl, 2,3-dihydroxypropyl, 2-methoxyethyl, methoxymethyl or 2- or 3-methoxypropyl.

5. The use according to claim 2 wherein the fumaric acid monoalkyl ester corresponds to the formula wherein Ri is as defined in claims 3 or 4 A is hydrogen, an alkali or alkaline earth metal cation or a physiologically compatible transition metal cation, preferably selected from Li', Na^, K , Mg"', Ca"\ Zn ~r,Fe''"r and Mn"T, and n is 1 or 2 and corresponds to the valence of A

6. The use according to any of the previous claims,characterised in thatthe fumaric acid derivative is one or more selected from the group comprising fumaric acid dimethyl ester, fumaric acid diethyl ester, fumaric acid methylethyl ester, methyl hydrogen fumarate, ethyl hydrogen fumarate, calcium methyl fumarate, calcium ethyl fumarate, magnesium methyl fumarate, magnesium ethyl fumarate, zinc methyl fumarate, zinc ethyl fumarate, iron methyl fumarate and iron ethyl fumarate and mixtures thereof.

7. The use according to claim 6,characterised in thatthe fumaric acid derivative is fumaric acid dimethyl ester (dimethyl fumarate).

8. The use of one or more fumaric acid derivatives for preparing a pharmaceutical composition for the therapy of diseases that may be influenced by NP-kappaB, selected from the group comprising: progressive systemic sclerodermia, osteochondritis syphilitica (Wegener's disease), cutis marmorata( livedo reticularis),Behcet disease, panarteritis, colitis ulcerosa, vasculitis, osteoarthritis, gout, arteriosclerosis, Reiter's disease, pulmonary granulomatosis, types of encephalitis, endotoxic shock (septic-toxic shock), sepsis, pneumonia, encephalomyelitis, anorexia ner vosa, hepatitis (acute hepatitis, chronic hepatitis, toxic hepatitis, alcohol-induced hepatitis, viral hepatitis, jaundice, liver insufficiency and cytomegaloviral hepatitis), Rennert T-lymphomatosis. mesangial nephritis, post-angioplastic restenosis, reperfusion syndrome, cytomegaloviral retinopathy, adenoviral diseases such as adenoviral colds, adenoviral pharin-goconjunctival fever and adenoviral ophthalmia, AIDS, Guillain-Barre syndrome, post-herpetic or post-zoster neuralgia, inflammatory demyelinising polyneuropathy, mononeuropathia multiplex, mucoviscidosis, Bechterew's disease, Barett oesophagus, EBV (Epstein-Barr virus) infec-tion, cardiac remodeling, interstitial cystitis, diabetes mellitus type II, human tumour radiosensitisation, multi-resistance of malignant cells to che- - motherapeutic agents (multidrug resistance in chemotherapy), granuloma annulare and cancers such as mamma carcinoma, colon carcinoma, melanoma, primary liver cell carcinoma, adenocarcinoma, kaposi's sarcoma, prostate carcinoma, leukaemia such as acute myeloid leukaemia, multiple myeloma (plasmocytoma), Burkitt lymphoma and Castleman rumour.

9. The use according to claim 8,characterised in thatthe fumaric acid derivative is selected from the group consisting of fumaric acid dialkyi esters and fumaric acid monoalkyl esters in the form of the free acid or a salt or mixtures thereof.

10. The use according to claim 9, where the fumaric acid dialkyi ester corre sponds to the formula wherein Rx and R2, which may be the same or different, independently represent a linear, branched, cyclic, saturated or unsaturated Ci _24 alkyl radical or a C5_2oaiyl radical and these radicals are optionally substituted with halogen (F, CI, Br, I), hydroxy, C]__4alkoxy, nitro or cyano.

11. The use according to one of the claims 9 and 10,characterised in thatthe radicals Rx and R2are methyl, ethyl, n-propyl, isopropyl, n-butyl, sec- butyl, t-butyl. pentyl. cyclopentyl, 2-ethylhexyl, hexyl, cyclohexyl, heptyl, cycloheptyl, octyl. vinyl, allyl, 2-hydroxyethyl. 2- or 3-hydroxypropyl. 2,3-dihydroxypropyl,2-methoxyethyl, methoxymethyl or 2- or 3-methoxypropyl.

12. The use according to claim 9 wherein the fumaric acid monoalkyl ester corresponds to the formula wherein Ri is as defined in claims 3 or 4 A is hydrogen, an alkali or alkaline earth metal cation or a physiologically compatible transition metal cation, preferably selected from Li<+>, Na<+>, K<+>, Mg<2+>, Ca2+, Zn2+, Fe<2+>and Mn2+, and n is 1 or 2 and corresponds to the valence of A

13. The use according to any of the claims 8 to 12,characterised in thatone dosage unit of the pharmaceutical composition contains an amount of fumaric acid derivative(s) corresponding to I to 500 mg, preferably 10 to 300 mg and most preferably 10 to 200 mg of fumaric acid.

14. The use according to any of the previous claims 8 to 13 for preparing a pharmaceutical composition for oral, parenteral, rectal, transdermal, dermal, nasal, pulmonal (inhalation) or ophthal administration, preferably for oral administration.

15. The use according to claim 14 where the pharmaceutical composition for oral administration is present in the form of unit dose tablets, micro-tablets, micro-pellets or granulate (said micro-tablets, micro-pellets or the granulate optionally being encapsulated or filled into sachets), capsules or solutions for drinking.

16. The use according to claim 15,characterised in thatthe solid dosage forms are provided with an enteric coating.

17. The use according to claim 8,characterised in thatthe dosage units of the pharmaceutical composition preferably contain either individually or in admixture: 10 to 500 mg of dialkyi fumarate, especially dimethyl fumarate and/or diethyl fumarate; 10 to 500 mg of calcium alkyl fumarate, especially calcium methyl fumarate and/or calcium ethyl fumarate 0 to 250 mg of zinc alkyl fumarate, especially zinc methyl fumarate and/or zinc ethyl fumarate, 0 to 250 mg of alkyl hydrogen fumarate, especially methyl hydrogen fumarate and/or ethyl hydrogen fumarate; and 0 to 250 mg magnesium alkyl fumarate, especially magnesium methyl fumarate and/or magnesium ethyl fumarate the total of the above-mentioned amounts corresponding to an equivalent of 10 to 500 mg, preferably 10 to 300 mg and most preferably 100 mg of fumaric acid.

18. The use according to one of the claims 15 or 16,characterised in thatthe composition is present in the form of micro-tablets or micro-pellets having a size of < 5000 um, preferably a size of 300 to 1000 urn for the pellets and 1000 to 2500 um for the micro-tablets. The present invention relates to the use of one or more fumaric acid derivatives as NF-kappaB inhibitor. At the same time, the present invention relates to the use of the fumaric acid derivatives for preparing a pharmaceutical composition for treating diseases that may be influenced by NF-kappaB.