HRP20010777A2 - INHIBITORS OF LFA-1 BINDING TO ICAMs AND USES THEREOF - Google Patents

INHIBITORS OF LFA-1 BINDING TO ICAMs AND USES THEREOF Download PDF

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HRP20010777A2
HRP20010777A2 HR20010777A HRP20010777A HRP20010777A2 HR P20010777 A2 HRP20010777 A2 HR P20010777A2 HR 20010777 A HR20010777 A HR 20010777A HR P20010777 A HRP20010777 A HR P20010777A HR P20010777 A2 HRP20010777 A2 HR P20010777A2
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sulfide
chlorophenyl
amino
chloro
dichlorophenyl
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Kerry Fowler
Mark Orme
Donald
Janet Adolphson
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Icos Corp
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Description

Područje izuma Field of invention

Izum je iz područja farmakologije. The invention is from the field of pharmacology.

Stanje tehnike State of the art

Antigen koji je povezan s funkcijom leukocita (LFA-1, CD11a/CD18) je leukocit-specifičan β2 integrin koji sudjeluje u stanica/stanica adheziji. Vezujuća aktivnost LFA-1 je bitna za ekstravazaciju leukocita od cirkulacije do mjesta ozljede pri upalnom odgovoru. Tri glavna liganda za koje se zna da vežu LFA-1; ICAM-1, ICAM-2 i ICAM-3, su međustanične adhezijske molekule koje imaju značajnu ulogu pri lokalizaciji leukocitne adhezije za endotelne stanice na mjestu ozljede. Nađeno je da ICAM-4 i ICAM-5 vežu LFA-1. Većina leukocita konstitutivno vrši ekspresiju LFA-1, ali vezanje liganda zahtijeva aktiviranje za koje se vjeruje da izaziva konformacijske promjene i povećava avidnost vezanja liganda. Primjerice, ICAM-1 ima normalnu ekspresiju na nižim razinama endotelija, međutim ozljedom izazvani medijatori upale dovode do povećane površinske ekspresije u stanicama na mjestu ozljede što, opet, izaziva lokaliziranu adheziju leukocita putem vezanja s aktiviranim LFA-1. Leukocyte function-related antigen (LFA-1, CD11a/CD18) is a leukocyte-specific β2 integrin that participates in cell/cell adhesion. The binding activity of LFA-1 is essential for the extravasation of leukocytes from the circulation to the site of injury in the inflammatory response. The three major ligands known to bind LFA-1; ICAM-1, ICAM-2 and ICAM-3 are intercellular adhesion molecules that play a significant role in the localization of leukocyte adhesion to endothelial cells at the site of injury. ICAM-4 and ICAM-5 were found to bind LFA-1. Most leukocytes constitutively express LFA-1, but ligand binding requires activation, which is believed to induce conformational changes and increase ligand binding avidity. For example, ICAM-1 has normal expression at lower levels of the endothelium, however injury-induced inflammatory mediators lead to increased surface expression in cells at the site of injury which, in turn, induces localized leukocyte adhesion via binding to activated LFA-1.

U strukturi LFA-1 nalazimo odvojene unutarstanične i izvanstanične domene za koje se vjeruje da sudjeluju i/ili reguliraju ICAM vezanje. Od posebnog je interesa područje u αL lancu od približno 200 aminokiselina, označeno kao I domena, koje je nađeno je svim β2 integrinima, kao i u većini ostalih proteina. Činjenice ukazuju na to da je I domena značajna za LFA-1 vezanje za ICAM-1 i 3. Primjerice, anti-LFA-1 blokirajuća monoklonska antitijela bila su mapirana za epitope unutar I domene. Nadalje, rekombinantni polipeptidni fragmenti I domene, pokazalo se, inhibiraju adheziju koju određuje integrin i vežu ICAM-1. Unutar I domene LFA-1 (i drugih proteina) postoji adhezijsko mjesto koje ovisi o jednom ionu (MIDAS) za koje se preferentno vežu ioni mangana ili magenzija. Vezanje nekog od ovih kationa je neophodno za interakciju liganda i vjeruje se da izaziva konformacijske promjene LFA-1 koje su neophodne za vezanje. Vezanje kationa dakle može biti regulacijski mehanizam koji je odgovoran za promjene u izvanstaničnoj leukocitnoj okolini. Ovu hipotezu potkrepljuju opažanja da vezanje kalcijeva iona ustvari inhibira LFA-1 interakciju s ICAM-1. Zaista, pretpostavljeno je da je neaktivna LFA-1 konformacija rezultat vezanja kalcija, te da je zamjena kalcijeva iona s magnezijevim neophodan stupanj za LFA-1 aktiviranje [Griggs, et al.. J. Biol. Chem. 173:22113-22119 (1998)]. Pokazalo se da i ostali čimbenici izazivaju LFA-1 aktiviranje, uključujući sudjelovanje receptora T-stanica, stimuliranje citokina te in vitro PMA stimuliranje. In the LFA-1 structure, we find separate intracellular and extracellular domains that are believed to participate in and/or regulate ICAM binding. Of particular interest is a region in the αL chain of approximately 200 amino acids, designated the I domain, which is found in all β2 integrins, as well as in most other proteins. Evidence suggests that domain I is important for LFA-1 binding to ICAM-1 and 3. For example, anti-LFA-1 blocking monoclonal antibodies were mapped to epitopes within domain I. Furthermore, recombinant polypeptide fragments of domain I have been shown to inhibit integrin-mediated adhesion and bind ICAM-1. Within domain I of LFA-1 (and other proteins) there is a single ion-dependent adhesion site (MIDAS) to which manganese or magnesium ions preferentially bind. Binding of one of these cations is necessary for ligand interaction and is believed to induce conformational changes in LFA-1 that are necessary for binding. Cation binding can therefore be a regulatory mechanism responsible for changes in the extracellular leukocyte environment. This hypothesis is supported by the observation that calcium ion binding actually inhibits LFA-1 interaction with ICAM-1. Indeed, it has been hypothesized that the inactive LFA-1 conformation is the result of calcium binding, and that the exchange of calcium ions with magnesium is a necessary step for LFA-1 activation [Griggs, et al.. J. Biol. Chem. 173:22113-22119 (1998)]. Other factors have also been shown to induce LFA-1 activation, including T-cell receptor involvement, cytokine stimulation, and in vitro PMA stimulation.

Konkretnim rječnikom, identificiranje LFA-1/ICAM vezujućih mjesta daje ciljeve za moduliranje leukocitnih upalnih odgovora. Izdvojena su brojna antitijela koja mogu izazvati LFA-1 aktiviranje (vidi, primjerice, Landis, et al., J. Cell Biol. 120:1519-1527 (1993)] ili, naprimjer, spriječiti ICAM-1 interakciju [vidi primjerice, Randi and Hogg, J. Biol. Chem. 169:12395-12398 (1994)]. Prethodno identificiranje anti-LFA-1 aktivirajućih antitijela koja prepoznaju višestruke i udaljene izvanstanične epitope ukazuje na postojanje više nego jednog regulacijskog područja, za koja se pretpostavlja da su neovisna o signaliziranju citoplazme. Lokaliziranje LFA-1 mjesta koja vežu ICAM-1 istraženo je uporabom kimernih LFA-1 podjediničnih proteina koji sadrže humane i mišje komponente (Huang and Springer, J. Biol. Chem. 170:19008-19016 (1995)]. Istraživanja su ukazala da su rezidue koje koordiniraju vezanjem kationa i rezidue koje su proksimalno bitne za vezanje ICAM-1 na relativno ravnom međusloju. Preciznije omeđenje izvanstaničnih regulacijskih područja i dodirnih točaka za ICAM-1 vezanje će omogućiti oblikovanje učinkovitih modulatora. In concrete terms, identifying LFA-1/ICAM binding sites provides targets for modulating leukocyte inflammatory responses. A number of antibodies have been identified that can induce LFA-1 activation (see, for example, Landis, et al., J. Cell Biol. 120:1519-1527 (1993)) or, for example, prevent ICAM-1 interaction [see, for example, Randi and Hogg, J. Biol. Chem. 169:12395-12398 (1994)]. The previous identification of anti-LFA-1 activating antibodies that recognize multiple and distant extracellular epitopes indicates the existence of more than one regulatory region, which is hypothesized to be independent of cytoplasmic signaling Localization of LFA-1 ICAM-1 binding sites was investigated using chimeric LFA-1 subunit proteins containing human and mouse components (Huang and Springer, J. Biol. Chem. 170:19008-19016 (1995)) Research has shown that the residues that coordinate cation binding and the residues that are proximally essential for ICAM-1 binding are on a relatively flat interface. More precise delineation of the extracellular regulatory regions and contact points for ICAM-1 binding will allow shaping the effects of these modulators.

Prema tome, postoje potreba u tehnici da se točno odrede regulacijska područja za proteine koji sudjeluju u upalnim odgovorima, a posebice za LFA-1 i ICAM-e koji vežu LFA-1. Određivanje tercijarne (ili kvarterne) strukture proteina može odrediti moguća regulacijska područja što omogućuje racionalno oblikovanje biološki sukladnih malenih molekula za terapeutsku i profilaktičku intervenciju za upalne bolesti. Također postoji potreba u tehnici da se odrede spojevi koji mogu inhibirati LFA-1 vezanje za ICAM-e koji se mogu koristiti pri tretiranju upalnih bolesti. Accordingly, there is a need in the art to pinpoint regulatory regions for proteins involved in inflammatory responses, particularly for LFA-1 and LFA-1 binding ICAMs. Determining the tertiary (or quaternary) structure of proteins can determine possible regulatory areas, which enables the rational design of biologically compatible small molecules for therapeutic and prophylactic intervention for inflammatory diseases. There is also a need in the art to identify compounds that can inhibit LFA-1 binding to ICAMs that can be used in the treatment of inflammatory diseases.

Sažetak izuma Summary of the invention

Ovaj izum odnosi se na spojeve koji se vežu za nova regulacijska mjesta u I domeni LFA-1, pa time inhibiraju vezanje LFA-1 za ICAM-e koji vežu LFA-1. Ovaj izum prema tome daje metode za reguliranje adhezije leukocita za endotelne stanice. Spojevi ovog izuma korisni su za tretiranje patoloških stanja, kao što su ona povezana s upalnim bolestima, autoimunim bolestima, tumorskim metastazama, odbacivanjem transplantata i oštećenjem reperfuzije. Konkretno, ovaj izum se odnosi na diaril sulfide opće formule (I), njihove farmaceutski prihvatljive soli ili prolijekove, te na uporabu diaril sulfida, posebice spojeva formule (I), u inhibiranju vezanja LFA-1 za ICAM koji veže LFA-1. This invention relates to compounds that bind to new regulatory sites in the I domain of LFA-1, thereby inhibiting the binding of LFA-1 to ICAMs that bind LFA-1. The present invention therefore provides methods for regulating the adhesion of leukocytes to endothelial cells. The compounds of this invention are useful for treating pathological conditions, such as those associated with inflammatory diseases, autoimmune diseases, tumor metastasis, transplant rejection and reperfusion impairment. In particular, this invention relates to diaryl sulfides of general formula (I), their pharmaceutically acceptable salts or prodrugs, and to the use of diaryl sulfides, especially compounds of formula (I), in inhibiting the binding of LFA-1 to ICAM that binds LFA-1.

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gdje su A i B, neovisno, arilne skiupine odabrane iz skupa kojega sačinjavaju 5- i 6-člani aormatski prsteni, uključujući i ne ograničavajući se samo na njih – fenil, tienil, furil, pirimidinil, imidazolil, pirazolil, piridil, pirazinil, pirolil i piridazinil; where A and B are, independently, aryl groups selected from the group consisting of 5- and 6-membered aromatic rings, including but not limited to - phenyl, thienyl, furyl, pyrimidinyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrrolyl and pyridazinyl;

R1, R2 i R3 su neovisno odabrani iz skupa kojega sačinjavaju vodik, R1, R2 and R3 are independently selected from the group consisting of hydrogen,

-Ra, gdje je Ra vodik ili alkilna skupina koja sadrži od jedan od šest lančastih ili razgranatih ugljikovih atoma (C1-6 alkil), -Ra, where Ra is hydrogen or an alkyl group containing one of six chain or branched carbon atoms (C1-6 alkyl),

-halogen, gdje je halogen Cl, F, Br ili I, -halogen, where halogen is Cl, F, Br or I,

-NRbRc, gdje su Rb i Rc neovisno H, C1-6 alkil ili –CH2-aril, -NRbRc, where Rb and Rc are independently H, C1-6 alkyl or –CH2-aryl,

-NO2, -NO2,

-C(=O)Ra, -C(=O)Ra,

-CN, -CN,

-perfluorRa, kao trifluormetil, -perfluoroRa, as trifluoromethyl,

-N-C(=O)Ra, -N-C(=O)Ra,

-(CH2)n-NRbRc, gdje je n cijeli broj od 1 do 6, -(CH2)n-NRbRc, where n is an integer from 1 to 6,

5- ili 6-člani heterociklički prsten, bilo alifatski ili aromatski, koji sadrži jedan ili više O, N ili S, proizvoljno supstituiranih, kao što su morfolino, i –S-aril, gdje je aril 5- ili 6-člani aromatski prsten, proizvoljno supstituiran; A 5- or 6-membered heterocyclic ring, either aliphatic or aromatic, containing one or more O, N or S, optionally substituted, such as morpholino, and -S-aryl, where aryl is a 5- or 6-membered aromatic ring , arbitrarily substituted;

a R4, R5 i R6, neovisno, odabrani su iz skupa kojega sačinjavaju and R 4 , R 5 and R 6 , independently, are selected from the set they comprise

vodik, hydrogen,

-Ra, -Ra,

-O-Ra, -O-Ra,

-halo, -halo,

-NRbRc, -NRbRc,

-NO2, -NO2,

-C(=O)Ra, -C(=O)Ra,

-CN, -CN,

-perfluorRa, -perfluoroRa,

-N-C(=O)Ra, -N-C(=O)Ra,

-(CH2)n-NRbRc, i -(CH2)n-NRbRc, i

- 5- ili 6-člani heterociklički prsten, alifatski ili aromatski, koji sadrži jedan ili više O, N ili S, proizvoljno supstituiran, - 5- or 6-membered heterocyclic ring, aliphatic or aromatic, containing one or more O, N or S, arbitrarily substituted,

-S-aril, ili gdje -S-aryl, or where

R4 i R5 su uzeti zajedno da sačinjavaju 5- ili 6-člani aromatski prsten, koji proizvoljno sadrži jedan ili više O, N ili S u prstenu, proizvoljno supstituiran. R 4 and R 5 are taken together to form a 5- or 6-membered aromatic ring, optionally containing one or more O, N or S in the ring, optionally substituted.

Primjeri novih negativnih regulatora vezanja LFA-1 za ICAM-e uključuju, ali nisu ograničeni na spojeve koji su prikazani u tablici I. Examples of novel negative regulators of LFA-1 binding to ICAMs include, but are not limited to, the compounds shown in Table I.

Tablica I Table I

Primjeri negativnih regulatora Examples of negative regulators

3-Klor-4-(2-klorfenilsulfanil)-fenilamin hidroklorid 3-Chloro-4-(2-chlorophenylsulfanyl)-phenylamine hydrochloride

4-Nitro-2-klorfenil-(2',3'-diklorfenil)-sulfid 4-Nitro-2-chlorophenyl-(2',3'-dichlorophenyl)-sulfide

3-Klor-4-(2-naftilsulfanil)-fenilamin hidroklorid 3-Chloro-4-(2-naphthylsulfanyl)-phenylamine hydrochloride

3-Klor-4-(2,3-diklorfenilsulfanil)-fenilamin hidroklorid 3-Chloro-4-(2,3-dichlorophenylsulfanyl)-phenylamine hydrochloride

3-Klor-4-(2,4,5-triklorfenilsulfanil)-fenilamin hidroklorid 3-Chloro-4-(2,4,5-trichlorophenylsulfanyl)-phenylamine hydrochloride

3-Klor-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 3-Chloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride

4-(Benzotiazol-2-ilsulfanil)-3-klor-fenilam~ne 4-(Benzothiazol-2-ylsulfanyl)-3-chloro-phenylamine

3-Klor-4-(1-klor-naftalen-2-ilsulfanil)-fenilamin 3-Chloro-4-(1-chloro-naphthalen-2-ylsulfanyl)-phenylamine

3-Metoksi-4-(2,3-diklorfenilsulfanil)-fenilamin 3-Methoxy-4-(2,3-dichlorophenylsulfanyl)-phenylamine

5-Amino-2-(2,3-diklorfenilsulfanil)-acetophenone hidroklorid 5-Amino-2-(2,3-dichlorophenylsulfanyl)-acetophenone hydrochloride

4-(2,3-diklorfenilsulfanil)-fenilamin 4-(2,3-dichlorophenylsulfanyl)-phenylamine

3-Klor-4-(1-naftilsulfanil)-fenilamin hidroklorid 3-Chloro-4-(1-naphthylsulfanyl)-phenylamine hydrochloride

3-Metil-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 3-Methyl-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride

1-Acetamido-3-klor-4-(2,3-diklorfenilsulfanil)-benzen 1-Acetamido-3-chloro-4-(2,3-dichlorophenylsulfanyl)-benzene

4-Metilamino-2,2',4'-triklordifenilsulfid 4-Methylamino-2,2',4'-trichlorodiphenylsulfide

3-Brom-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 3-Bromo-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride

3-Hidroksi-4-(2,3-diklorfenilsulfanil)-fenilamin hidroklorid 3-Hydroxy-4-(2,3-dichlorophenylsulfanyl)-phenylamine hydrochloride

6-Klor-5-(2,4-diklorfenilsulfanil)-1 H-benzimidazol 6-Chloro-5-(2,4-dichlorophenylsulfanyl)-1H-benzimidazole

4-Amino-2-klorfenil-(2'4'-dimetilfenil)-sulfid hidroklorid 4-Amino-2-chlorophenyl-(2'4'-dimethylphenyl)-sulfide hydrochloride

2,5-Diklor-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 2,5-Dichloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride

4-Amino-2-klorfenil-(2'-metil-4'-klorfenil)-sulfid 4-Amino-2-chlorophenyl-(2'-methyl-4'-chlorophenyl)-sulfide

4-Amino-2-klorfenil-(2',4'-difluorfenil)-sulfid hidroklorid 4-Amino-2-chlorophenyl-(2',4'-difluorophenyl)-sulfide hydrochloride

4-Amino-2-klorfenil-(2',4',6'-triklorfenil)-sulfid 4-Amino-2-chlorophenyl-(2',4',6'-trichlorophenyl)sulfide

4-Amino-2-klorfenil-(2'-amino-4'-klorfenil)-sulfid 4-Amino-2-chlorophenyl-(2'-amino-4'-chlorophenyl)-sulfide

4-Amino-2-klorfenil-(2'-klor-4'-nitrofenil)-sulfid 4-Amino-2-chlorophenyl-(2'-chloro-4'-nitrophenyl)-sulfide

4-Amino-2-klorfenil-(2'-nitro-4'-klorfenil)-sulfid 4-Amino-2-chlorophenyl-(2'-nitro-4'-chlorophenyl)-sulfide

4-Amino-2-klorfenil-(3',4'-diklorfenil)-sulfid 4-Amino-2-chlorophenyl-(3',4'-dichlorophenyl)-sulfide

4-Amino-2-klorfenil-2-(3-klor-5-trifluormetilpiridil)-sulfid 4-Amino-2-chlorophenyl-2-(3-chloro-5-trifluoromethylpyridyl)-sulfide

Bis-(4,4'-diamino-2,2'-diklorfenil)-sulfid Bis-(4,4'-diamino-2,2'-dichlorophenyl)-sulfide

4-Amino-2-klorfenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-chlorophenyl-(2',4'-dichlorophenyl)-sulfide

4-Amino-2-klorfenil-(4'-acetamido-2'-klorfenil)-sulfid 4-Amino-2-chlorophenyl-(4'-acetamido-2'-chlorophenyl)-sulfide

4-Amino-2-klorfenil-6-(5-nitrokinolino)-sulfid 4-Amino-2-chlorophenyl-6-(5-nitroquinolino)-sulfide

4-Amino-2-klorfenil-(4'-dimetilamino-2'-klorfenil)-sulfid 4-Amino-2-chlorophenyl-(4'-dimethylamino-2'-chlorophenyl)-sulfide

2-Klor-4-amino-S-metilaminofenil-(2',4'-diklorfenil)-sulfid 2-Chloro-4-amino-S-methylaminophenyl-(2',4'-dichlorophenyl)-sulfide

2-Klor-4-amino-5-N-morfolinofenil-(2',4'-diklorfenil)-sulfid 2-Chloro-4-amino-5-N-morpholinophenyl-(2',4'-dichlorophenyl)-sulfide

4-Amino-2-trifluormetilfenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-trifluoromethylphenyl-(2',4'-dichlorophenyl)-sulfide

4-Amino-2-klorfenil-2-(5-nitro-3-brom)-pyridine sulfid 4-Amino-2-chlorophenyl-2-(5-nitro-3-bromo)-pyridine sulfide

4-Aminometil-2-klorfenil-(2', 4'-diklorfenil)-sulfid 4-Aminomethyl-2-chlorophenyl-(2', 4'-dichlorophenyl)-sulfide

4,5-Diklor-2-(2,4-diklorfenilsulfanil)-fenilamin 4,5-Dichloro-2-(2,4-dichlorophenylsulfanyl)-phenylamine

3,5-Diklor-4-(2,4-diklorfenilsulfanil)-fenilamin 3,5-Dichloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine

2,3-Diklor-4-(2,4-diklorfenilsulfanil)-fenilamin 2,3-Dichloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine

4-Amino-2-fluorfenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-fluorophenyl-(2',4'-dichlorophenyl)-sulfide

5-Amino-3-klorfenil-(2',4'-diklorfenil)-sulfid 5-Amino-3-chlorophenyl-(2',4'-dichlorophenyl)-sulfide

3-Klor-4-( 1-klor-nafthalen-2-ilsulfanil)-fenilamin 3-Chloro-4-(1-chloro-naphthalen-2-ylsulfanyl)-phenylamine

1-(3-Nitro-4-fenilsulfanil-fenil)-etanon 1-(3-Nitro-4-phenylsulfanyl-phenyl)-ethanone

1-(3-Nitro-4-fenilsulfanil-fenil)-etanon oksim 1-(3-Nitro-4-phenylsulfanyl-phenyl)-ethanone oxime

5-Trifluormetil-2-fenilsulfanil-benzonitril 5-Trifluoromethyl-2-phenylsulfanyl-benzonitrile

1-(3,5-diklorfenil)-3-fenilsulfanil-pirolidin-2,5-dion 1-(3,5-dichlorophenyl)-3-phenylsulfanyl-pyrrolidine-2,5-dione

Bis-2,4,6-Trinitrofenil-sulfid Bis-2,4,6-Trinitrophenyl-sulphide

2-Metil-1-(2-o-tolilsulfanil-fenil)-1H pirol 2-Methyl-1-(2-o-tolylsulfanyl-phenyl)-1H pyrrole

3-[2-(4-Klor-2-nitro-fenilsulfanil)-fenilamino-3H isobenzofuran-1-one 3-[2-(4-Chloro-2-nitro-phenylsulfanyl)-phenylamino-3H isobenzofuran-1-one

4-(Benzotiazol-2-ilsulfanil)-3-klor-fenilamin 4-(Benzothiazol-2-ylsulfanyl)-3-chloro-phenylamine

2-Nitro-4-klorfenil-(2'aminofenil)-sulfid 2-Nitro-4-chlorophenyl-(2'aminophenyl)-sulfide

6-Amino-2-klorfenil-(4'-metilfenil)-sulfid 6-Amino-2-chlorophenyl-(4'-methylphenyl)-sulfide

4-Nitrofenil-(2'-klorfenil)-sulfid 4-Nitrophenyl-(2'-chlorophenyl)sulfide

2, 4-Dinitrofenil-(4'-klorfenil)-sulfid 2, 4-Dinitrophenyl-(4'-chlorophenyl)-sulfide

4-Aminofenil-(2'-klorfenil)-sulfid 2, 4-Aminophenyl-(2'-chlorophenyl)-sulfide 2,

4-Diaminofenil-(4'-izopropilfenil)-sulfid 4-Diaminophenyl-(4'-isopropylphenyl)-sulfide

4-Nitro-2-klorfenil-(2',3'-diklorfenil)-sulfid 4-Nitro-2-chlorophenyl-(2',3'-dichlorophenyl)-sulfide

4-Amino-2-klorfenil-2-(5-nitro-3-brom)-piridin-sulfid 4-Amino-2-chlorophenyl-2-(5-nitro-3-bromo)-pyridine-sulfide

Spojevi kojima odgovara strukturna formula (I) mogu se prirediti sintetskim metodama ili metaboličkim procesima. Priređivanje spojeva metaboličkim procesima uključuje in vivo i in vitro procese. Podrazumijevaju se i farmaceutske smjese koje sadrže spojeve ovog izuma. Compounds corresponding to the structural formula (I) can be prepared by synthetic methods or metabolic processes. Preparation of compounds by metabolic processes includes in vivo and in vitro processes. Pharmaceutical mixtures containing the compounds of this invention are also understood.

Izumom su definirane metode inhibiranja LFA-1 vezanja za ICAM-e koji vežu LFA-1, koje uključuju kontakt LFA-1 s diaril sulfidom, te poželjno spojem formule 1. Slično, izumom su definirane metode za inhibiranje adhezije leukocita za endotelne stanice koje uključuju stupanj kontakta leukocita s ekspresijom LFA-1 s diaril sulfidom, te poželjno spojem strukturne formule (I). Izum također obuhvaća metode za tretiranje upalnih bolesti koje uključuju stupnjeve primjene sisavcu određene količine farmaceutske smjese ovog izuma koja je dovoljna za inhibiranje vezanja LFA-1 za svoje prirodne ligande koji kompetiraju s ICAM-1 ili ICAM-3 za vezanje s LFA-1. Izumom su također obuhvaćene metode za tretiranje upalnih bolesti koje nastaju nakon LFA-1 vezanja za svoje prirodne ligande koji kompetiraju s ICAM-1 ili ICAM-3 vezanjem za LFA-l, koje uključuju primjenu sisavcu kojemu je to potrebno spoja koji kompetira s 3-klor-4-(1-klornaftalen-2-ilsulfanil)-fenilaminom za vezanje na LFA-1 u količini koja je dovoljna da se inhibira vezanje za prirodne ligande LFA-1. Nadalje, izumom su definirane metode za poboljšanje patoloških stanja koja su povezana s LFA-1 vezanjem za ICAM koji veže LFA-1, koje uključuju primjenu osobi kojoj je to potrebno učinkovite količine diaril sulfida, te poželjno spoja strukturen formule (I) za inhibiranje LFA-1 vezanja za ICAM. The invention defines methods of inhibiting LFA-1 binding to ICAMs that bind LFA-1, which include contacting LFA-1 with diaryl sulfide, and preferably with a compound of formula 1. Similarly, the invention defines methods for inhibiting leukocyte adhesion to endothelial cells that include the degree of contact of leukocytes with LFA-1 expression with diaryl sulfide, and preferably with the compound of structural formula (I). The invention also encompasses methods for treating inflammatory diseases comprising the steps of administering to a mammal an amount of a pharmaceutical composition of the invention sufficient to inhibit the binding of LFA-1 to its natural ligands that compete with ICAM-1 or ICAM-3 for binding to LFA-1. Also encompassed by the invention are methods for treating inflammatory diseases resulting from LFA-1 binding to its native ligands that compete with ICAM-1 or ICAM-3 binding to LFA-1, comprising administering to a mammal in need thereof a compound that competes with 3- with chloro-4-(1-chloronaphthalen-2-ylsulfanyl)-phenylamine to bind to LFA-1 in an amount sufficient to inhibit binding to native LFA-1 ligands. Furthermore, the invention defines methods for ameliorating pathological conditions associated with LFA-1 binding to ICAM that binds LFA-1, which include administering to a person in need of it an effective amount of diaryl sulfide, and preferably a compound of formula (I) to inhibit LFA -1 bindings to ICAM.

Primjeri inhibitora ovog izuma uključuju, ali nisu ograničeni na spojeve koji su navedeni u tablici I. Examples of inhibitors of the present invention include, but are not limited to, the compounds listed in Table I.

Izumom je također obuhvaćena primjena spoja ovog izuma pri proizvodnji medikamenta za tretiranje patoloških stanja koja su povezana s LFA-1 koji se veže za ICAM-1. The invention also covers the application of the compound of this invention in the production of medication for the treatment of pathological conditions associated with LFA-1 that binds to ICAM-1.

Ovim izumom su također obuhvaćene metode za identificiranje negativnog regulatora LFA-1 koji se veže za svoje prirodne ligande koji kompetiraju s ICAM-1 ili ICAM-3 za vezanje s LFA-1, što uključuje sljedeće stupnjeve: Also encompassed by the present invention are methods for identifying a negative regulator of LFA-1 that binds to its native ligands that compete with ICAM-1 or ICAM-3 for binding to LFA-1, comprising the following steps:

a) dovođenje u međusobni kontakt LFA-1 s aktivatorom vezanja LFA-1; a) bringing LFA-1 into mutual contact with the LFA-1 binding activator;

b) mjerenje LFA-1 vezanja s prirodnim ligandom u prisutnosti i odsutnosti ispitivanog spoja; i b) measurement of LFA-1 binding with the natural ligand in the presence and absence of the test compound; and

c) identificiranje spoja koji se ispituje kao inhibitora kada se smanjeno LFA-1 vezanje za ligand uočava u prisutnosti ispitivanog spoja. U jednom aspektu, aktivator je kristalno ljubičasto (crystal violet). c) identifying the test compound as an inhibitor when reduced LFA-1 ligand binding is observed in the presence of the test compound. In one aspect, the activator is crystal violet.

Realizacija izuma s primjerima izvođenja Realization of the invention with examples of execution

Vrijednost IC50 spoja definira se kao ona koncentracija spoja koje je potrebna da se postigne 50% inhibiranje biološke aktivnosti koja je od interesa. Kako se ovdje rabi, negativni regulator je spoj koji je karakteriziran s IC50 vrijednošću za inhibiranje LFA-1 vezanja za prirodni ligand. Negativni regulatori LFA-1 definirani su kao oni koji imaju IC50 vrijednost manju od oko 200 µM, manju od oko 100 µM, manju od oko 50 µM, te poželjno od oko 0,05 µM do oko 40 µM. The IC50 value of a compound is defined as that concentration of the compound required to achieve 50% inhibition of the biological activity of interest. As used herein, a negative regulator is a compound characterized by an IC50 value for inhibiting LFA-1 binding to a native ligand. Negative regulators of LFA-1 are defined as having an IC50 value of less than about 200 µM, less than about 100 µM, less than about 50 µM, and preferably from about 0.05 µM to about 40 µM.

Pojam “farmaceutski prihvatljivi nosač” kako se ovdje rabi odnosi se na one prolijekove spojeva ovog izuma koji su pogodni za uporabu u kontaktu s životnjama primateljima i za koje su nepoželjna toksičnost, nadražaj i alergijski odgovor u razumnom odnosu boljitak/rizik, te koji su učinkoviti za predviđenu uporabu. The term "pharmaceutically acceptable carrier" as used herein refers to those prodrugs of the compounds of this invention which are suitable for use in contact with recipient animals and for which undesirable toxicity, irritation and allergic response are in a reasonable benefit/risk ratio, and which are effective for the intended use.

Pojam “prolijek” kako se ovdje rabi, odnosi se na spojeve koji podliježu brzoj transformaciji in vivo matičnog spoja gore navedene formule, primjerice hidrolizom. Iscrpna diskusija može se naći u Higuchi, et al., Prodrugs as Novel Delivery Systems, vol. 14 A.C.S.D. Symposium Series, te u Roche (ed), Bioreversible Carriers in Drug Design, American Phannaceutical Association i Pergamon Press, 1987, a oboje je ovdje uključeno kao referenca. Oblikovanje prolijeka općenito je razmotreno u Hardma, et al., (Eds), Goodman & Gilman's The Pharmacological Basis of Therapeutics, Ninth Edition, New York, New York (1996), pp. 11-16. Ukratko, nakon primjene lijeka slijedi uklanjanje iz tijela ili neka biološka transformacija, čime se biološka aktivnost lijeka smanjuje ili uklanja. Alternativno, proces biološke transformacije može rezultirati metaboličkim sporednim produktima koji su sami metabolički aktivniji ili jednako aktivni kao i lijek koji je početno primijenjen. Bolje razumijevanje ovih bioloških promjena omogućuje oblikovanje tzv. “prolijekova” koji, nakon biološke transformacije, postaju biološki aktivni na nekom drugom mjestu. Prolijekovi su prema tome farmakološki neaktivni spojevi koji su prevedeni u biološki aktivne metabolite. U nekim oblicima, prolijekovi ostaju biološki aktivni tijelom hidrolize, primjerice, esterske ili amidne veze, pri čemu se često uvodi ili izlaže funkcionalna skupina na prolijeku. Tako promijenjeni lijek može također reagirati s endogenim spojem tako da nastane u vodi topljivi konjugat koji dodatno povećava farmakološka svojstva spoja, primjerice kao rezultat povećanog poluživota u krvotoku. The term "prodrug" as used herein refers to compounds that are subject to rapid in vivo transformation of the parent compound of the above formula, for example by hydrolysis. A comprehensive discussion can be found in Higuchi, et al., Prodrugs as Novel Delivery Systems, vol 14 A.C.S.D. Symposium Series, and in Roche (ed), Bioreversible Carriers in Drug Design, American Phannaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference. Prodrug formulation is generally discussed in Hardma, et al., (Eds), Goodman & Gilman's The Pharmacological Basis of Therapeutics, Ninth Edition, New York, New York (1996), pp. 11-16. In short, drug administration is followed by removal from the body or some biological transformation, which reduces or removes the drug's biological activity. Alternatively, the process of biological transformation may result in metabolic byproducts that are themselves metabolically more active or as active as the drug initially administered. A better understanding of these biological changes enables the formation of the so-called "prodrugs" which, after biological transformation, become biologically active in another place. Prodrugs are therefore pharmacologically inactive compounds that have been converted into biologically active metabolites. In some forms, prodrugs remain biologically active by hydrolysis of, for example, ester or amide bonds, often introducing or exposing a functional group on the prodrug. The drug thus changed can also react with the endogenous compound to form a water-soluble conjugate that further increases the pharmacological properties of the compound, for example as a result of an increased half-life in the bloodstream.

Kao druga mogućnost, prolijekovi mogu biti tako oblikovani da podliježu kovalentnoj promjeni na funkcionalnoj skupini s, primjerice, glukuronskom kisleinom, sulfatom, glutationom, aminokiselinama ili acetatom. Nastali konjugat može biti deaktiviran i izlučen u žuč, podvrgnut enzimskom cijepanju ili ostati jači od svog matičnog spoja. Visokomolekularni konjugati mogu također biti izlučeni u žuč, podvrgnuti enzimskom cijepanju i vraćeni natrag u optok, čime se djelotvorno povećava biološki vrijeme poluživota izvorno primijenjenog spoja. Alternatively, prodrugs may be designed to undergo covalent modification of the functional group with, for example, glucuronic acid, sulfate, glutathione, amino acids, or acetate. The resulting conjugate can be deactivated and excreted in the bile, subjected to enzymatic cleavage or remain more potent than its parent compound. High molecular weight conjugates may also be secreted into the bile, undergo enzymatic cleavage, and be returned to the circulation, thereby effectively increasing the biological half-life of the originally administered compound.

Spojevi ovog izuma mogu postojati kao stereoizomeri s asimetričnim ili kiralnim centrima. Stereoizomeri su označeni bilo “S” ili “R”, ovisno o rasporedu supstituenata oko kiralnog ugljikova atoma. Ovim izumom obuhvaćene su smjese stereoizomera. Stereoizomeri uključuju enantiomere, diastereomere i njihove smjese. Pojedinačni stereoizomeri spojeva ovog izuma mogu se prirediti sintetski oz komercijalno raspoloživih tvari koje imaju asimetrične ili kiralne centre ili priređivanjem racemičkih smjesa, nakon čega slijede tehnike odvajanja ili razlučivanja, koje su poznate u tehnici. Metode razlučivanja su (1) dodavanje smjese enantiomera za kiralnu pkmoćnu tvar, odvajanje dobivene smjese pomoćšu rekristaliziranja ili kromatografije, te oslobađanje optički čistog produkta od pomoćne tvari; (2) nastajanje soli korištenjem optički aktivnog sredstva za razlučivanje, i (3) izravno odvajanje smjese optičkih enantiomera na kiralnim kromatografskim kolonama. The compounds of this invention may exist as stereoisomers with asymmetric or chiral centers. Stereoisomers are designated either “S” or “R”, depending on the arrangement of substituents around the chiral carbon atom. Mixtures of stereoisomers are covered by this invention. Stereoisomers include enantiomers, diastereomers, and mixtures thereof. Individual stereoisomers of the compounds of this invention can be prepared synthetically from commercially available materials having asymmetric or chiral centers or by preparing racemic mixtures, followed by separation or resolution techniques known in the art. The methods of resolution are (1) addition of a mixture of enantiomers for a chiral active substance, separation of the resulting mixture using recrystallization or chromatography, and liberation of the optically pure product from the auxiliary substance; (2) salt formation using an optically active resolving agent, and (3) direct separation of a mixture of optical enantiomers on chiral chromatographic columns.

Spojevi ovog izuma uključuju, ali nisu ograničeni na one koji su obuhvaćeni općom formulom (I) i spojeve koji su navedeni u tablici I. The compounds of this invention include, but are not limited to, those covered by general formula (I) and the compounds listed in Table I.

Izumom se također dobivaju farmaceutske smjese koje sadrže jedan ili više spojeva ovog izuma, koje također sadrže i farmaceutski prihvatljiv nosač ili razrjeđivač. The invention also provides pharmaceutical mixtures containing one or more compounds of this invention, which also contain a pharmaceutically acceptable carrier or diluent.

Izumom su također obuhvaćene metode inhibiranja LFA-1 vezanja za ICAM koji veže LFA-1 što uključuje stupanj dovođenja u dodir LFA-1, ili ICAM-vezujući fragment, s negativnim regulatorskim spojem; pri čemu negativni regulator veže LFA-1 αL polipeptid ili njegov fragment, na mjestu koje je odabrano iz skupa kojega sačinjava konformacija koja veže diaril sulfid ili vezujuće mjesto koje je definirano s Ile259, Leu298, Ile235, Val157, Leu161 i Ile306 humanog LFA-1 αL polipeptida i LFA-1 domene koja veže 3-klor-4-(1-klor-naftalen-2-ilsulfanil)-fenilamin koja ima strukturu koja je prije opisana. Alternativno, vezujuće mjesto negativnog regulatora na LFA-1 definirano je aminokiselinskim reziduama Ile259, Leu298, Ile235, Val157, Leu161, Ile306, Leu302,Tyr257, Leu132, Val233, Val130 i Tyr166. U sljedećoj alternativi, vezujuće mjesto negativnog regulatora na LFA-1 definirano je aminokiselinskim reziduama Lys287, Leu298, Ile259, Leu302, Ile235, Val157, Tyr257, Lys305, Leu161, Leu132, Val233, Ile255, Val130, Tyr166, Ile306, Phe134, Phe168, Phe153, Tyr307, Val308, Ile309, Thr231, Glu284, Phe285, Glu301, Met154, Ile237, Ile150 i Leu295. Vezujuće mjesto LFA-1 regulatora opisano je u pratećoj američkoj patentnoj prijavi koja je naslovljena "Vezujuće mjesto LFA-1 regulatora i njegovo korištenje”, podnesenoj 2. travnja 1999. br. 27866/35375, serijski broj 09/285,477, koja je ovdje u cijelosti uključena kao referenca. U jednoj realizaciji, metode ovog izuma obuhvaćaju korištenje stanica koje vrše ekspresiju bilo LFA-1 ili ICAM. Also encompassed by the invention are methods of inhibiting LFA-1 binding to LFA-1-binding ICAM comprising the step of contacting LFA-1, or an ICAM-binding fragment, with a negative regulatory compound; wherein the negative regulator binds the LFA-1 αL polypeptide or fragment thereof, at a site selected from the group consisting of the diaryl sulfide binding conformation or the binding site defined by Ile259, Leu298, Ile235, Val157, Leu161 and Ile306 of human LFA-1 of an αL polypeptide and an LFA-1 3-chloro-4-(1-chloro-naphthalen-2-ylsulfanyl)-phenylamine-binding domain having the structure previously described. Alternatively, the negative regulator binding site on LFA-1 is defined by amino acid residues Ile259, Leu298, Ile235, Val157, Leu161, Ile306, Leu302,Tyr257, Leu132, Val233, Val130 and Tyr166. In the following alternative, the negative regulator binding site on LFA-1 is defined by amino acid residues Lys287, Leu298, Ile259, Leu302, Ile235, Val157, Tyr257, Lys305, Leu161, Leu132, Val233, Ile255, Val130, Tyr166, Ile306, Phe134, Phe168. Phe153, Tyr307, Val308, Ile309, Thr231, Glu284, Phe285, Glu301, Met154, Ile237, Ile150 and Leu295. The LFA-1 regulator binding site is described in a companion US patent application entitled "LFA-1 Regulator Binding Site and Use," filed Apr. 2, 1999, No. 27866/35375, Serial No. 09/285,477, which is hereby incorporated by reference. incorporated by reference in its entirety In one embodiment, the methods of the present invention comprise the use of cells expressing either LFA-1 or ICAM.

U metodama gdje se vrši ekspresija u stanici jednog od vezujućih partnera, drugi vezujući partner je bilo pročišćen ili izdvojen, u tekućem uzorku (pročišćen, djelomično pročišćen ili sirov) koji je uzet od neke osobe ili u staničnom lizatu. Izumom su također obuhvaćene metode gdje se vrši ekspresija u stanici i LFA-1 i ICAM. Ekspresija LFA-1 i ICAM vezujućih partnera može biti na istom staničnom tipu ili na različitim tipovima stanica. Poželjno, ekspresija LFA-1 polipeptida je na leukocitima, tj. limfocitima, monocitima ili granulocitima, dok je ekspresija ICAM polipeptida na endotelnim stanicama. In methods where expression is carried out in a cell of one of the binding partners, the other binding partner is either purified or isolated, in a liquid sample (purified, partially purified or crude) taken from an individual or in a cell lysate. The invention also covers methods where both LFA-1 and ICAM are expressed in a cell. Expression of LFA-1 and ICAM binding partners can be on the same cell type or on different cell types. Preferably, the expression of the LFA-1 polypeptide is on leukocytes, ie, lymphocytes, monocytes or granulocytes, while the expression of the ICAM polypeptide is on endothelial cells.

Izumom su također definirane metode za inhibiranje adhezije leukocita za endotelne stanice, koje uključuju stupanj dovođenja u dodir navedenih leukocita s negativnim regulatorom LFA-1 vezanja za ICAM koji veže LFA-1, pri čemu je negativan regulator vezanja nekog LFA-1 regulacijskog mjesta odabran iz skupine koju sačinjava konformacija koja veže diaril sulfid ili vezujuće mjesto koje je definirano s Ile259, Leu298, Ile235, Val157, Leu161 i Ile306 humanog LFA-1 αL polipeptida ili LFA-1 domene koja veže 3-klor-4-(1-klor-naftalen-2-ilsulfanil)-fenilamin. Alternativno, vezujuća konformacija diaril sulfida definirana je aminokiselinskim reziduama kao što je prije opisano. Podrazumijevaju se in vivo i in vitro metode. The invention also defines methods for inhibiting the adhesion of leukocytes to endothelial cells, which include the step of contacting said leukocytes with a negative regulator of LFA-1 binding to ICAM that binds LFA-1, wherein the negative regulator of binding of an LFA-1 regulatory site is selected from group formed by the diaryl sulfide-binding conformation or the binding site defined by Ile259, Leu298, Ile235, Val157, Leu161 and Ile306 of the human LFA-1 αL polypeptide or the LFA-1 domain that binds 3-chloro-4-(1-chloro- naphthalen-2-ylsulfanyl)-phenylamine. Alternatively, the binding conformation of diaryl sulfide is defined by amino acid residues as previously described. This includes in vivo and in vitro methods.

Ovim izumom su također definirane metode za poboljšanje patološkog stanja koje je posljedica LFA-1 vezanja za ICAM, koje obuhvaćaju primjenu osobi kojoj je potrebno negativnog regulatora LFA-1 vezanja za ICAM u količini koja je učinkovita za inhibiranje LFA-1 vezanja za ICAM, pri čemu je negativan regulator vezanja za LFA-1 regulacijsko mjesto odabran iz skupa kojega sačinjava konformacija koja veže diaril sulfid ili mjesto koje je definirano s Ile259, Leu298, Ilei235, Val157, Leu161 i Ile306 humanog LFA-1 ili humane LFA-1 domene koji vežu spoj 3-klor-4-(1-klor-naftalen-ilsulfanil)-fenilamin. The present invention also defines methods for ameliorating a pathological condition resulting from LFA-1 binding to ICAM, comprising administering to a subject in need thereof a negative regulator of LFA-1 binding to ICAM in an amount effective to inhibit LFA-1 binding to ICAM, at wherein the negative regulator binding to the LFA-1 regulatory site is selected from the group consisting of the diaryl sulfide binding conformation or the site defined by Ile259, Leu298, Ilei235, Val157, Leu161 and Ile306 of human LFA-1 or human LFA-1 binding domains compound 3-chloro-4-(1-chloro-naphthalen-ylsulfanyl)-phenylamine.

U poželjnoj realizaciji, metode ovog izuma uključuju uporabu diarilnog sulfidnog spoja za inhibiranje vezanja LFA-1 za ICAM. Poželjne metode uključuju uporabu spoja opće formule (I), njegove farmaceutski prihvatljive soli ili njegova prolijeka, kao što je prije opisano. In a preferred embodiment, the methods of the present invention include the use of a diaryl sulfide compound to inhibit the binding of LFA-1 to ICAM. Preferred methods include the use of a compound of general formula (I), a pharmaceutically acceptable salt thereof or a prodrug thereof, as previously described.

Terapeutske metode Therapeutic methods

U opsegu u kojem je adhezija leukocita za endotelne stanice povezanom s patološkim stanjem, izumom su definirane metode za poboljšanje patološkog stanja koje je povezano s akumuliranjem leukocita zbog LFA-1 vezanja za neki ICAM koji veže LFA-1 koje uključuju stupanj primjene osobi kojoj je to potrebno određene količine inhibitora LFA-1 vezanja za ICAM koja je učinkovita glede inhibiranja LFA-1 vezanja za ICAM, pri čemu se navedeni inhibitor veže za LFA-1 na mjestu na kojem se nalaze aminokiselinske rezidue Ile259, Leu298, Ile235, Val157, Leu161 i Ile306. Primjeri takvih medicinskih stanja uključuju, bez ograničenja, upalne bolesti, autoimune bolesti, reperefuzijske ozljede, infarkt miokarda, udar, hemoragični šok, presađivanje organa, i slično. Metode ovog izuma omogućuju poboljšanje cijelog niza patoloških stanja, uključujući primjerice (bez ograničenja) respiratorni stres sindrom, sindrom višestruke ozljede organa sekundarno do septikemije, sindrom višestruke ozljede organa sekundarno do traume; reperfuzijska ozljeda tkiva, akutni glomeluronefritis, reaktivni artritis, dermatoza s akutnim upalnim komponentama, moždani udar, toplinska ozljeda, Crohn-ova bolest; nekrotizirajući enterokolitis, sindrom povezan s granulocitnom transfuzijom, citokinom izazvana toksičnost, te bolesti koje su upravljane T-stanicama. To the extent that adhesion of leukocytes to endothelial cells is associated with a pathological condition, the invention defines methods for ameliorating a pathological condition associated with the accumulation of leukocytes due to LFA-1 binding to an ICAM that binds LFA-1, which include the degree of administration to a subject requires a specific amount of an inhibitor of LFA-1 binding to ICAM that is effective in inhibiting LFA-1 binding to ICAM, wherein said inhibitor binds to LFA-1 at the site where amino acid residues Ile259, Leu298, Ile235, Val157, Leu161 and Ile306. Examples of such medical conditions include, without limitation, inflammatory diseases, autoimmune diseases, reperfusion injury, myocardial infarction, stroke, hemorrhagic shock, organ transplantation, and the like. The methods of the present invention enable amelioration of a variety of pathological conditions, including for example (without limitation) respiratory stress syndrome, multiple organ injury syndrome secondary to septicemia, multiple organ injury syndrome secondary to trauma; tissue reperfusion injury, acute glomerulonephritis, reactive arthritis, dermatosis with acute inflammatory components, stroke, thermal injury, Crohn's disease; necrotizing enterocolitis, granulocyte transfusion-related syndrome, cytokine-induced toxicity, and T-cell driven diseases.

Upalno stanično aktiviranje te suvišni ili neregulirani citokini (npr. TNFα i IL-1β) također su obuhvaćeni bolestima kao što je reumatoidni artritis, osteoartritis, giht artritis, spondilitis, oftalmopatija povezana sa štitnom žlijezdom, Behcet-ova bolest, sepsa, septički šok, ednotoksični šok, gram negativna sepsa, gram pozitivna sepsa, sindrom toksičnog šoka, astma, kronični bronhitis, alergijski respiratorni distres sindrom, kronična plućna upalna bolest kao što je kronična ostruktivna plućna bolest, silikoza, plućna sarkoidoza, reperfuzijska ozljeda mikarda, mozda i ekstremiteta, fibroza, cistična fibroza, keloidno formiranje, nastajanje ožiljaka, ateroskleroza, bolesti odbacivanja transplantata, kronični glomerulonefritis, lupus, upalna trbušna bolest kao što je ulcerativni kolitis, proliferativna limfocitna bolest kao što je leukemija, te upalne dermatoze, kao što su atopički dermatitis, psorijaza, urtikarija, uveitis. Inflammatory cell activation and excess or dysregulated cytokines (eg, TNFα and IL-1β) are also implicated in diseases such as rheumatoid arthritis, osteoarthritis, gouty arthritis, spondylitis, thyroid-related ophthalmopathy, Behcet's disease, sepsis, septic shock, ednotoxic shock, gram negative sepsis, gram positive sepsis, toxic shock syndrome, asthma, chronic bronchitis, allergic respiratory distress syndrome, chronic pulmonary inflammatory disease such as chronic obstructive pulmonary disease, silicosis, pulmonary sarcoidosis, reperfusion injury of the mycardium, brain and extremities, fibrosis, cystic fibrosis, keloid formation, scarring, atherosclerosis, transplant rejection diseases, chronic glomerulonephritis, lupus, inflammatory bowel disease such as ulcerative colitis, proliferative lymphocytic disease such as leukemia, and inflammatory dermatoses such as atopic dermatitis, psoriasis , urticaria, uveitis.

Ostala stanja koja su karakterizirana povišenom razinom citokina uključuju ozljedu mozga zbog umjerene traume (vidi J. Neurotrauma, 12, pp. 1035-1043 (1995); J. Clin. Invest., 91, pp. 1421-1428 (1993)), kardiomiopatije, kao što je kongestivan srčani ispad (vidi Circulation, 97, pp. 1340-1341 (1998)), kaheksija, kaheksija sekundarno do infekcije ili malignosti, kaheksija sekundarno do sindroma imune deficijencije (AIDS), ARC (kompleks povezan s AIDS-om), mialgije groznice zbog infekcije, moždana malarija, osteoporoza i bolesti koštane resorpcije, keloidno formiranje, nastajanje ožiljnog tkiva i pireksija. Other conditions characterized by elevated cytokine levels include mild traumatic brain injury (see J. Neurotrauma, 12, pp. 1035-1043 (1995); J. Clin. Invest., 91, pp. 1421-1428 (1993)), cardiomyopathies, such as congestive heart failure (see Circulation, 97, pp. 1340-1341 (1998)), cachexia, cachexia secondary to infection or malignancy, cachexia secondary to immunodeficiency syndrome (AIDS), ARC (AIDS-related complex om), fever myalgias due to infection, cerebral malaria, osteoporosis and bone resorption diseases, keloid formation, formation of scar tissue and pyrexia.

Sposobnost negativnih regulatora ovog izuma za tretiranje artritisa može se demonstrirati modelom mišjeg kolagenom izazvanog artritisa [Kakimoto, et al. Immunol. 142:326-337 (1992)], modelom štakorova kolagenom izazvanog artritisa [Knoerzer, et al., Toxical Pathol. 25:13-19 (1997)], modelom štakorova adjuvantnog artritisa [Halloran, et al., Arthritis Rheum 39:810-819 (1996)], modelom štakorova stijekom streptokokne stanice izazvanog artritisa [Schimmer, et al., J. Immunol. 160:1466-1477 (1998)], ili modelom SCID-mišjeg humanog reumatoidnog artritisa [Oppenheimer-Marks, et al., J. Clin. Invest 101:1261-1272 (1998)]. The ability of the negative regulators of the present invention to treat arthritis can be demonstrated in a murine model of collagen-induced arthritis [Kakimoto, et al. Immunol. 142:326-337 (1992)], a rat model of collagen-induced arthritis [Knoerzer, et al., Toxical Pathol. 25:13-19 (1997)], a rat model of adjuvant arthritis [Halloran, et al., Arthritis Rheum 39:810-819 (1996)], a rat model of streptococcal cell-induced arthritis [Schimmer, et al., J. Immunol . 160:1466-1477 (1998)], or the SCID-mouse model of human rheumatoid arthritis [Oppenheimer-Marks, et al., J. Clin. Invest 101:1261-1272 (1998)].

Sposobnost negativnih regulatora za tretiranje Lyme artritisa može se demonstrirati sukladno metodi Gross, et al., Science, 218:703-706, (1998). Sposobnost negativnih regulatora za tretiranje astme može se demonstrirati modelom mišje alergijske astme sukladno metodi Wegner, et al., Science, 247:456-459, (1990), ili modelom mišje nealergijske astme sukladno metodi Bloemen, et al., Am. J. Respir. Crit. Care Med. 153:521529 (1996). The ability of negative regulators to treat Lyme arthritis can be demonstrated according to the method of Gross, et al., Science, 218:703-706, (1998). The ability of negative regulators to treat asthma can be demonstrated by a mouse model of allergic asthma according to the method of Wegner, et al., Science, 247:456-459, (1990), or by a mouse model of non-allergic asthma according to the method of Bloemen, et al., Am. J. Respir. Crit. Care Med. 153:521529 (1996).

Sposobnost negativnih regulatora za tretiranje upalne ozljede pluća može se demonstrirati modelom kisikom-izazvane mišje ozljede pluća sukladno metodi Wegner, et al., Lung, 170:267-279, (1992), modelom imunim kompleksom izazvane mišje ozljede pluća sukladno metodi Mulligan, et al., J. Immunol., 154:1350-1363, (1995), ili modelom kiselinom izazvane mišje ozljede pluća sukladno metodi Nagase, et al., Am. J. Respir. Crit. Care Med., 154:504-510, (1996). The ability of negative regulators to treat inflammatory lung injury can be demonstrated by the oxygen-induced mouse lung injury model according to the method of Wegner, et al., Lung, 170:267-279, (1992), the immune complex-induced mouse lung injury model according to the method of Mulligan, et al. al., J. Immunol., 154:1350-1363, (1995), or by an acid-induced murine lung injury model according to the method of Nagase, et al., Am. J. Respir. Crit. Care Med., 154:504-510, (1996).

Sposobnost negativnih regulatora za tretiranje upalnog gihta može se demonstrirati modelom mišjeg kemijski izazvanog kolitisa sukladno metodi Bennett, et al., J. Pharmacol. Exp. Ther.,180:988-1000, (1997). The ability of negative regulators to treat inflammatory gout can be demonstrated by a murine model of chemically induced colitis according to the method of Bennett, et al., J. Pharmacol. Exp. Ther., 180:988-1000, (1997).

Sposobnost negativnih regulatora za tretiranje autoimunog dijabetesa može se demonstrirati modelom NOD miša sukladno metodi Hasagawa, et al., Int. Immunol. 6:831-838 (1994), ili modelom mišjeg streptomicin-izazvanog dijabetesa sukladno metodi Herrold, et al., Cell Immunol. 157:489-500, (1994). The ability of negative regulators to treat autoimmune diabetes can be demonstrated with the NOD mouse model according to the method of Hasagawa, et al., Int. Immunol. 6:831-838 (1994), or by a mouse model of streptomycin-induced diabetes according to the method of Herrold, et al., Cell Immunol. 157:489-500, (1994).

Sposobnost negativnih regulatora za tretiranje upalne ozljede jetre može se demonstrirati modelom ozljede mišje jetre sukladno metodi Tanaka, et al., J. Immunol., 15I:5088-5095, (1993). The ability of negative regulators to treat inflammatory liver injury can be demonstrated by a mouse liver injury model according to the method of Tanaka, et al., J. Immunol., 15I:5088-5095, (1993).

Sposobnost negativnih regulatora za tretiranje upalne glomerularne ozljede može se demonstrirati modelom nefritisa štakorova nefrotoksična seruma sukladno metodi Kawasaki, et al., J. Immunol., 150:1074-1083 (1993). The ability of negative regulators to treat inflammatory glomerular injury can be demonstrated with a rat nephrotoxic serum nephritis model according to the method of Kawasaki, et al., J. Immunol., 150:1074-1083 (1993).

Sposobnost negativnih regulatora za tretiranje radijacijski-izazvanog enteritisa može se demonstrirati modelom štakorova abdominalnog ozračenja sukladno metodi Panes, et al., Gastroerrterology, 108:1761-1769 (1995). The ability of negative regulators to treat radiation-induced enteritis can be demonstrated in a rat model of abdominal irradiation according to the method of Panes, et al., Gastroenterology, 108:1761-1769 (1995).

Sposobnost negativnih regulatora za tretiranje radijacijskog preumonitisa može se demonstrirati modelom mišjeg plućnog ozračenja sukladno metodi Hallahan, et al., Proc. Natl. Acad. Sci (USA), 94:6432-6437 (1997). The ability of negative regulators to treat radiation premonitis can be demonstrated with a murine lung irradiation model according to the method of Hallahan, et al., Proc. Natl. Acad. Sci (USA), 94:6432-6437 (1997).

Sposobnost negativnih regulatora za tretiranje reperfuzijske ozljede može se demonstrirati u izoliranog srca sukladno metodi Tamiya, et al., Immunopharmacology, 29:53-63 (1995), ili u anesteziranog psa sukladno modelu Hartman, et al., Cardiovasc. Res. 30:47-54 (1995). The ability of negative regulators to treat reperfusion injury can be demonstrated in the isolated heart according to the method of Tamiya, et al., Immunopharmacology, 29:53-63 (1995), or in the anesthetized dog according to the model of Hartman, et al., Cardiovasc. Crisp. 30:47-54 (1995).

Sposobnost negativnih regulatora za tretiranje plućne reperfuzijske ozljede može se demonstrirati modelom reperfuzijske ozljede pluća štakora sukladno metodi deMeester, et al., Transplantation, 62:1477-1485 (1996), ili modelom plućnog edema kunića sukladno metodi Horgan, et al., Am. J. Physiol. 261:H1578-H1584 (1991). The ability of negative regulators to treat lung reperfusion injury can be demonstrated by the rat lung reperfusion injury model according to the method of deMeester, et al., Transplantation, 62:1477-1485 (1996), or the rabbit pulmonary edema model according to the method of Horgan, et al., Am. J. Physiol. 261:H1578-H1584 (1991).

Sposobnost negativnih regulatora za tretiranje može se demonstrirati modelom moždanog embolijskog udara kunića sukladno metodi Bowes, et al., Exp. Neurol., 119:215-219 (1993), modelom ishemije/reperfuzije srednje moždane arterije štakora metodom Chopp, et al., Stroke, 2.5:869875 (1994), ili modelom povratne ishemije kralježnice kunića sukladno metodi Clark et al., Neurosurg., 75:623-627 (1991). Sposobnost negativnih regulatora za tretiranje moždanog spazma krvnih žila može se demonstrirati modelom eksperimentalnog vazospazma štakora sukladno metodi Oshiro, et al., Stroke, 18:2031-2038 (1997). The ability of negative regulators to treat can be demonstrated in a rabbit cerebral embolic stroke model according to the method of Bowes, et al., Exp. Neurol., 119:215-219 (1993), the rat middle cerebral artery ischemia/reperfusion model by the method of Chopp, et al., Stroke, 2.5:869875 (1994), or the rabbit spinal cord ischemia-reperfusion model according to the method of Clark et al., Neurosurg. ., 75:623-627 (1991). The ability of negative regulators to treat cerebral vasospasm can be demonstrated by a rat model of experimental vasospasm according to the method of Oshiro, et al., Stroke, 18:2031-2038 (1997).

Sposobnost negativnih regulatora za tretiranje periferne arterijske okluzije može se demonstrirati modelom ishemije/reperfuzije štakorova skeletna mišića sukladno metodi Gute, et al., Mol. Cell Biochem., 179:169-187 (1998). The ability of negative regulators to treat peripheral arterial occlusion can be demonstrated by a rat skeletal muscle ischemia/reperfusion model according to the method of Gute, et al., Mol. Cell Biochem., 179:169-187 (1998).

Sposobnost negativnih regulatora za tretiranje odbacivanja organa može se demonstrirati modelom odbacivanja mišjeg srca sukladno metodi Isobe, et al., Science,155:1125-1127 (1992), modelom bubrežne kapsule mišje štitine žlijezde sukladno metodi Talento, et al., Transplantation, 55:418-422 (1993), modelom prihvaćanja bubrega cinomolškog majmuma sukladno metodi Cosimi, et al., J. Immunol., 144:4604-4612 (1990), modelom prihvaćanja živa štakora sukladno metodi Nakao, et al., Muscle Nerve, 18:93-102 (1995), modelom prihvaćanja mišje kože sukladno metodi Gorczynski and Wojcik, J. Immunol. 152:2011-2019, (1994), modelom prihvaćanja mišje rožnice sukladno metodi He, et al., Ophtalmol. Vis. Sci., 35:3218-3225 (1994), ili modelom ksenogenskog presađivanja stanice gušterače sukladno metodi Zeng, et al., Transplantation, 58:681-689 (1994). The ability of negative regulators to treat organ rejection can be demonstrated by the mouse heart rejection model according to the method of Isobe, et al., Science, 155:1125-1127 (1992), by the renal capsule model of the mouse thyroid gland according to the method of Talento, et al., Transplantation, 55 :418-422 (1993), cynomolgus monkey kidney uptake model according to the method of Cosimi, et al., J. Immunol., 144:4604-4612 (1990), live rat uptake model according to the method of Nakao, et al., Muscle Nerve, 18:93-102 (1995), mouse skin acceptance model according to the method of Gorczynski and Wojcik, J. Immunol. 152:2011-2019, (1994), by the mouse cornea acceptance model according to the method of He, et al., Ophthalmol. Height. Sci., 35:3218-3225 (1994), or by a model of xenogeneic pancreatic cell transplantation according to the method of Zeng, et al., Transplantation, 58:681-689 (1994).

Sposobnost negativnih regulatora za tretiranje bolesti odbacivanja organa (GVHD) može se demonstrirati modelom mišjeg letalnog GVHD sukladno metodi Harning, et al., Transplantation, 52:842-845 (1991). The ability of negative regulators to treat organ rejection disease (GVHD) can be demonstrated in a mouse model of lethal GVHD according to the method of Harning, et al., Transplantation, 52:842-845 (1991).

Sposobnost negativnih regulatora za tretiranje može se demonstrirati modelom metastaze humanog limfoma (u miševa) sukladno modelu Aoudjit, et al., J. Immunol., 161:2333-2338, (1998). The ability of negative regulators to treat can be demonstrated by a human lymphoma metastasis model (in mice) according to Aoudjit, et al., J. Immunol., 161:2333-2338, (1998).

Farmaceutske smjese Pharmaceutical mixtures

Ovim izumom također su definirane farmaceutske smjese koje sadrže diaril sulfid formuliran zajedno s jednim ili više farmaceutski prihvatljivih nosača. The present invention also defines pharmaceutical compositions containing diaryl sulfide formulated together with one or more pharmaceutically acceptable carriers.

Farmaceutske smjese ovog izuma mogu se primijeniti ljudima ili životinjama bilo kojim pogodnim putem. Primjerice, smjese se mogu primijeniti oralno, rektalno, parenteralno, intracisterno, intravaginalno, intraperitonealno, lokalno (kao prašci, pomade ili kapi), bukalno ili nazalno. Pojam “parenteralna” primjena kako se ovdje rabi označuje načine primjene koji uključuju intravensku, intramuskularnu, intraperitonealnu, intrasternalnu, intratekalnu, supkutanu ili intraartikularnu injekciju i infuziju. The pharmaceutical compositions of the present invention may be administered to humans or animals by any convenient route. For example, the compositions can be administered orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (as powders, ointments or drops), buccally or nasally. The term "parenteral" administration as used herein refers to routes of administration including intravenous, intramuscular, intraperitoneal, intrasternal, intrathecal, subcutaneous or intra-articular injection and infusion.

Farmaceutske smjese ovog izuma za parenteralnu injekciju obuhvaćaju farmaceutski prihvatljive sterilne vodene ili nevodene otopine, disperzije, suspenzije ili emulzije kao i sterilne praške za rekonstituiranje s sterilne otopine ili disperzije za injiciranje neposredno prije uporabe. Primjeri pogodnih vodenih ili nevodenih nosača, razrjeđivala, otapala ili posrednika uključuju vodu, etanol, polioli (kao što su glicerol, propilenglikol, polietilenglikol i slično), te njihove odgovarajuće smjese, biljna ulja (kao što je maslinovo ulje) te injicirajući organski esteri kao što je etil oleat. Odgovarajuća protočnost može se održati, primjerice, uporabom materijala za prevlake kao što je lecitin, zadržavanjem odgovarajuće veličine čestica u slučaju disperzija, te uporabom površinski aktivnih tvari. The pharmaceutical compositions of this invention for parenteral injection include pharmaceutically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions as well as sterile powders for reconstitution from a sterile solution or dispersion for injection immediately before use. Examples of suitable aqueous or non-aqueous carriers, diluents, solvents or mediators include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol and the like), and their respective mixtures, vegetable oils (such as olive oil) and injectable organic esters such as which is ethyl oleate. Adequate flowability can be maintained, for example, by using coating materials such as lecithin, by maintaining the appropriate particle size in the case of dispersions, and by using surfactants.

Ove smjese mogu također sadržavati adjuvante kao što su konzervansi, sredstva za vlaženje, sredstva za emulgiranje i raspršivanje. Sprječavanje djelovanja mikroorganizama može se provesti uključivanjem različitih antibakterijskih i antigljivičnih sredstava, primjerice parabena, klorbutanola, fenol sorbične kiseline i slično. Također može biti poželjno da se uključe izotonična sredstva kao što su, primjerice, šećeri, natrijev klorid i slična. Produžena apsorpcija injicirajućeg farmaceutskog oblika može se postići uključivanjem sredstva koje odgađa apsorpciju kao što je aluminijev monostearat i želatina. These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying and dispersing agents. Preventing the action of microorganisms can be carried out by including various antibacterial and antifungal agents, for example paraben, chlorobutanol, phenol sorbic acid and the like. It may also be desirable to include isotonic agents such as, for example, sugars, sodium chloride and the like. Prolonged absorption of the injectable pharmaceutical form can be achieved by including an absorption delaying agent such as aluminum monostearate and gelatin.

U nekim slučajevima, da bi se produžio učinak lijeka, poželjno je usporiti apsorpciju lijeka nakon supkutane ili intramuskularne injekcije. To se može provesti uporabom tekuće suspenzije kristalnog ili amorfnog materijala koji je slabo topljiv u vodi. Brzina apsorpcije lijeka onda ovisi o brzini njegova otapanja koje opet može ovisiti o veličini kristala i kristalnom obliku. Alternativno, odgođena asporpcija lijeka koji je parenteralno primijenjen može se realizirati otapanjem ili raspršenjem lijeka u uljnom nosaču. In some cases, in order to prolong the effect of the drug, it is desirable to slow down the absorption of the drug after subcutaneous or intramuscular injection. This can be done by using a liquid suspension of crystalline or amorphous material that is poorly soluble in water. The rate of drug absorption then depends on the rate of its dissolution, which in turn may depend on the crystal size and crystal form. Alternatively, delayed absorption of parenterally administered drug can be achieved by dissolving or dispersing the drug in an oil carrier.

Injicirajući depo oblici izrađuju se oblikovanjem mikroenkapsuliranih matrica lijeka u biološki razgradivim polimerima kao što su polilaktid-poliglikolid. Ovisno o odnosu lijeka prema polimeru i prirodi konkretnog polimera koji se rabi, može se kontrolirsti brzina otpuštanja lijeka. Primjeri drugih biološki razgradivih polimera uključuju poli(ortoestere) i poli(anhidride). Depo injicirajuće formulacije mogu se također prirediti stavljanjem lijeka u liposome ili mikroemulzije koje su sukladne s tjelesnim tkivom. Injectable depot forms are made by forming microencapsulated drug matrices in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of the drug to the polymer and the nature of the specific polymer used, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations can also be prepared by encapsulating the drug in liposomes or microemulsions that are compatible with body tissue.

Formulacije za injiciranje mogu se primjerice sterilizirati filtriranjem kroz bakterijski filter ili filter koji zadržava viruse, ili ugrađivanjem sredstava za sterilizaciju u obliku sterilnih čvrstih smjesa koje se mogu otopiti ili raspršiti u sterilnoj vodi ili drugom sterilnom mediju za injiciranje neposredno prije uporabe. Injectable formulations can be sterilized, for example, by filtering through a bacterial or virus-retaining filter, or by incorporating sterilizing agents in the form of sterile solids that can be dissolved or dispersed in sterile water or other sterile injection media immediately prior to use.

Čvrsti dozirajući oblici za oralnu primjenu uključuju kapsule, tablete, pilule, praške i granule. U takvih čvrstim dozirajućim oblicima, aktivan spoj se miješa s bar jednim inertnim, farmaceutski prihvatljivim ekscipijentom ili nosačem kao što je natrijev citrat i dikalcijev fosfat i/ili punila kao što su škrob, laktoza, saharoza, glukoza, manitol i silicijeva kiselina, (b) veziva kao što su, primjerice, karboksimetilceluloza, gume (npr. alginati, akacija), želatine, polivinilpirolidon i saharoza, (c) vlažila kao što su glicerol, (d) sredstva za razgradnju kao što su agar-agar, kalcijev karbonat, krumpirov ili tapioka škrob, alginska kiselina, neki silikati i natrijev karbonat, (e) sredstva koja zadržavaju tekuće stanje kao što je parafin, (f) ubrzivači apsorbiranja kao što su kvarterni amonijevi spojevi, (g) sredstva za vlaženje kao što su primjerice cetilni alkohol i glicerol monostearat, (h) apsorbensi kao što su kaolin i bentonitna glina, te (i) lubrikanti kao što su talk, kalcijev stearat, magnezijev stearat, čvrsti polietilen glikoli, natrijev lauril sulfat i njihove smjese. U slučaju kapsula, tableta i pilula, dozirajući oblik može također sadržavati sredstva za puferiranje. Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate and dicalcium phosphate and/or fillers such as starch, lactose, sucrose, glucose, mannitol and silicic acid, (b ) binders such as, for example, carboxymethylcellulose, gums (e.g. alginates, acacia), gelatins, polyvinylpyrrolidone and sucrose, (c) wetting agents such as glycerol, (d) disintegrants such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, some silicates and sodium carbonate, (e) agents that retain the liquid state such as paraffin, (f) absorption accelerators such as quaternary ammonium compounds, (g) wetting agents such as cetyl alcohol and glycerol monostearate, (h) absorbents such as kaolin and bentonite clay, and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also contain buffering agents.

Čvrste smjese sličnog tipa mogu se također upotrebljavati kao punila u mekim ili tvrdim kapsulama korištenjem takvih ekscipijenata kao laktoza ili mliječni šećer, kao i visokomolekularnih polietileglikola i slično. Solid mixtures of a similar type can also be used as fillers in soft or hard capsules using such excipients as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like.

Čvrsti oblici za doziranje kao što su tablete, dražeje, kapsule, pilule i granule mogu se prirediti s prevlakama kao što su enteričke prevlake ili druge prevlake koje su dobro poznate u tehnici farmaceutskog formuliranja. Oni mogu proizvoljno sadržavati sredstva koja daju neprozirnost i mogu također biti takvog sastava koji otpušta aktivne sastojke isključivo ili djelomično u crijevnom traktu, proizvoljno na odgođeni način. Primjerni materijali uključuju polimere koji su topljivosti koja ovisi o pH, kakvi se javljaju pod imenom Eudragit®. Primjeri ugrađenih smjesa koje se mogu koristiti su polimerne tvari i voskovi. Solid dosage forms such as tablets, dragees, capsules, pills and granules may be prepared with coatings such as enteric coatings or other coatings well known in the art of pharmaceutical formulation. They may optionally contain agents that provide opacity and may also be of such a composition that releases the active ingredients exclusively or partially in the intestinal tract, optionally in a delayed manner. Exemplary materials include polymers of pH-dependent solubility, such as occur under the name Eudragit®. Examples of embedded compounds that can be used are polymeric substances and waxes.

Aktivni spojevi mogu također biti u mikroenkapsuliranom obliku ako je to odgovarajuće, s jednim ili više prije navedenih ekscipijenata. The active compounds may also be in microencapsulated form if appropriate, with one or more of the aforementioned excipients.

Tekući dozirajući oblici za oralnu primjenu uključuju farmaceutski prihvatljive emulzije, otopine, suspenzije, sirupe i eliksire. Uz aktivne spojeve, tekući dozirajući oblici mogu sadržavati inertna razrjeđivala koja su poznata u tehnici, kao što su primjerice voda i druga otapala, sredstva za otapanje i emulgiranje kao što su etilni alkohol, etil acetat, etil karbonat, benzilni alkohol, benzil benzoat, propilenglikol, 1,3-butilenglikol, dimetilformamid, ulja (konkretno ulje orašca, kukuruzno, ulje klica, maslinovo ulje, dabrovo i sezamovo ulje), glicerol, tetrahidrofurfurilni alkohol, polietilenglikolni i esteri masnih kiselina sorbitana, te njihove smjese. Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs. In addition to active compounds, liquid dosage forms may contain inert diluents known in the art, such as water and other solvents, solubilizing and emulsifying agents such as ethyl alcohol, ethyl acetate, ethyl carbonate, benzyl alcohol, benzyl benzoate, propylene glycol , 1,3-butylene glycol, dimethylformamide, oils (specifically walnut, corn, germ oil, olive oil, beaver and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and sorbitan fatty acid esters, and their mixtures.

Uz inertna razrjeđivala, oralne smjese mogu također sadržavati adjuvante kao što su sredstva za vlaženje, emulgiranje i suspendiranje, te sredstva za boju, miris i okus. In addition to inert diluents, oral formulations may also contain adjuvants such as wetting, emulsifying and suspending agents, and coloring, fragrance and flavoring agents.

Suspenzije, uz aktivne spojeve, mogu sadržavati sredstva za raspršivanje kao što su, primjerice, etoksilirani izostearilni alkoholi, polioksietilen sorbitol i sorbitan esteri, mikrokristalna celuloza, aluminijev metahidroksid, bentonit, agar-agar i tragakant, te njihove smjese. Suspensions, in addition to active compounds, may contain dispersing agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and their mixtures.

Smjese za rektalnu i vaginalnu primjenu mogu poželjno sadržavati supozitorije koje se mogu prirediti miješanjem spojeva ovog izuma s odgovarajućim nenadražujućim ekscipijentima ili nosačima kao što su kakao maslac, polietilen glikol ili vosak za supozitorije koji je čvrst na sobnoj temperaturi ali je tekuć pri tjelesnoj temperaturi te se prema tome tali u rektalnoj ili vaginalnoj šupljini oslobađajući aktivan spoj. Compositions for rectal and vaginal administration may preferably contain suppositories which may be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or suppository wax which is solid at room temperature but liquid at body temperature and therefore, it dissolves in the rectal or vaginal cavity, releasing the active compound.

Spojevi ovog izuma mogu se također primijeniti u obliku liposoma. Kao što je poznato u tehnici, liposomi se općenito ozvode iz fosfolipida ili ostalih lipidnih tvari. Liposomi nastaju iz mono- ili višelamelnih hidrtiranih tekućih kristala koji su raspršeni u vodenom mediju. Može se koristiti bilo koji netoksični, fiziološki prihvatljiv i metabolizirajući lipid koji može izgrađivati liposome. Ove smjese u liposomnom obliku, uz spoj ovog izuma, mogu sadržavati spoj ovog izuma, stabilizatore, konzervanse, ekscipijente i slično. Poželjni lipidi su fosfolipidi i fosfatidil kolini (lecitini), prirodni i umjetni. Metode dobivanja lecitina su poznate u tehnici. Vidi, primjerice, Prescott, Ed., Methods in Cell Biology, Vo1ume XN, Academic Press, New York, N.Y. ( 1976), p. 33 i dalje. The compounds of the present invention may also be administered in the form of liposomes. As known in the art, liposomes are generally derived from phospholipids or other lipid substances. Liposomes are formed from mono- or multi-lamellar hydrated liquid crystals that are dispersed in an aqueous medium. Any non-toxic, physiologically acceptable and metabolizable lipid capable of forming liposomes can be used. These mixtures in liposomal form, in addition to the compound of this invention, may contain the compound of this invention, stabilizers, preservatives, excipients and the like. Preferred lipids are phospholipids and phosphatidyl cholines (lecithins), natural and artificial. Methods of obtaining lecithin are known in the art. See, for example, Prescott, Ed., Methods in Cell Biology, Volume XN, Academic Press, New York, N.Y. (1976), pp. 33 et seq.

Spojevi ovog izuma mogu se koristiti u obliku farmaceutski prihvatljivih soli koje su izvedene iz anorganskih ili organskih kiselina. The compounds of this invention may be used in the form of pharmaceutically acceptable salts derived from inorganic or organic acids.

“Farmaceutski prihvatljivom soli” smatraju se one soli koje su, unutar dosega jasnog medicinskog rasuđivanja, pogodne za uporabu u dodiru s tkivima čovjeka i nižih životinja bez nepoželjne toksičnosti, nadražaja, alergijskog odgovora i slično, te odgovara razumnom odnosu boljitak/rizik. Farmaceutski prihvatljive soli su dobro poznate u tehnici. Primjerice, S. M. Berge, et al., opisuje detaljno farmaceutski prihvatljive soli u J. Pharmaceutical Sciences, 66:1 (1977). Doli se mogu prirediti in situ tijekom konačnog odvajanja i pročišćavanja spoja izuma ili odvojeno, reakcijom funkcijske skupine slboodne baze s odgovarajućom kiselinom. Reprezentativne kisele adicijske soli uključuju, ali nisu ograničene na acetat, adipat, alginat, citrat, aspartat, benzoat, benzensulfonat, bisulfat, butirat, kamforat, kamforolsulfonat, diglukonat, glicerofosfat, hemisulfat, heptanoat, heksanoat, fumarat hidroklorid, hidrobromid, hidrojodid, 2-hidroksietansulfonat (izotionat), laktat, maleat, metansulfonat, nikotinat, 2-naftalensulfonat, oksalat, pamoat, pektinat, persulfat, 3-fenilpropionat, pikrat, pivalat, propionat, sukcinat, tartrate, tiocijanat, fosfat, glutamat, bikarbonat, p-toluensulfonat i undekanoat. Primjeri kiselina koje se mogu koristiti za dobivanje farmaceutski prihvatljivih kiselih adicijskih soli uključuju anroganske kiseline kao što su klorovodična kiselina, bromovodična kiselina, sumporna kiselina i fosforna kiselina te takve organske kiseline kao što su oksalna kiselina, maleinska kiselina, jantarna kiselina i limunska kiselina. "Pharmaceutically acceptable salts" are considered those salts which, within the scope of clear medical reasoning, are suitable for use in contact with the tissues of humans and lower animals without undesirable toxicity, irritation, allergic response and the like, and correspond to a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge, et al., describes in detail pharmaceutically acceptable salts in J. Pharmaceutical Sciences, 66:1 (1977). Doles can be prepared in situ during the final separation and purification of the compound of the invention or separately, by reacting the functional group of a free base with an appropriate acid. Representative acid addition salts include, but are not limited to acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, fumarate hydrochloride, hydrobromide, hydroiodide, 2 -hydroxyethanesulfonate (isothionate), lactate, maleate, methanesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, phosphate, glutamate, bicarbonate, p- toluenesulfonate and undecanoate. Examples of acids that can be used to prepare pharmaceutically acceptable acid addition salts include inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid, and such organic acids as oxalic acid, maleic acid, succinic acid, and citric acid.

Bazične skupine koje sadrže dušik mogu biti kvarternizirane s takvim sredstvima kao što su metil, etil, propil i butil kloridi, bromidi i jodidi; dialkil sulfati kao dimetil, dietil, dibutil i diamil sulfati; dugolančani halogenidi kao što su decil, lauril, miristil i stearil kloridi, bromidi i jodidi; arilalkil halogenidi kao što su benzil i fenetil bromidi i ostali. Time se dobivaju u vodi ili u ulju topljivi produkti ili dispergirani produkti. Nitrogen-containing basic groups can be quaternized with such agents as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates such as dimethyl, diethyl, dibutyl and diamyl sulfates; long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; arylalkyl halides such as benzyl and phenethyl bromides and others. This results in water- or oil-soluble products or dispersed products.

Bazične adicijske soli mogu se prirediti in situ tijekom konačnog izdvajanja i pročišćavanja spoja ovog izuma reakcijom vrste koja sadrži karboksilnu kiselinu s odgovarajućom bazom kao što je hidorksid, karbonat ili bikarbonat farmaceutski prihvatljivog metalnog kationa ili amonija ili organskog primarnog , sekundarnog ili tercijarnog amina. Farmaceutski prihvatljive bazične adicijske soli uključuju, ali nisu isključivo ograničene na katione alkalnih metala ili zemnoalkalnih metala kao što su soli litija, natrija, kalija, kalcija, magnezija il aluminija i slično, te netoksične kvarterne amonijeve ili aminske katione uključujući amonij, tetrametilamonij, tetraetilamonij, metilamin, dimetilamin, trimetilamin, trietilamin, dietilamin, etilamin i slično. Ostali reprezentativni organski amini koji su korisni za dobivanje baznih adicijskih soli uključuju etilendiamin, atanolamin, dietanolamin, piperidin, piperazin i slično. Base addition salts can be prepared in situ during the final isolation and purification of a compound of this invention by reacting a carboxylic acid-containing species with an appropriate base such as a hydroxide, carbonate or bicarbonate of a pharmaceutically acceptable metal cation or ammonium or an organic primary, secondary or tertiary amine. Pharmaceutically acceptable base addition salts include, but are not exclusively limited to alkali metal or alkaline earth metal cations such as lithium, sodium, potassium, calcium, magnesium, and aluminum salts and the like, and non-toxic quaternary ammonium or amine cations including ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, diethylamine, ethylamine and the like. Other representative organic amines that are useful for making base addition salts include ethylenediamine, ethanolamine, diethanolamine, piperidine, piperazine, and the like.

Dozirajući oblici za lokalnu primjenu spoja ovog izuma uključuju praške, sprejeve, pomade i inhalante. Aktivan spoj miješa se u sterilnim uvjetima s farmaceutski prihvatljivim nosačem te s eventualno potrebnim konzervansima, puferima ili propelantima. Formulacija za očnu primjenu, očne pomade, prašci i otopine također su unutar dosega ovog izuma. Dosage forms for topical application of the compounds of this invention include powders, sprays, pomades and inhalants. The active compound is mixed under sterile conditions with a pharmaceutically acceptable carrier and with any necessary preservatives, buffers or propellants. Ocular formulation, ophthalmic ointments, powders and solutions are also within the scope of this invention.

Stvarna razina doziranja aktivnih sastojaka u farmaceutskim smjesama može varirati da se postigne određena količina aktivnog spoja (spojeva) koja je djelotvorna da se postigne određeni terapeutski odgovor za određenog bolesnika, smjesu i način primjene. Odabrana razina doziranja ovisi o aktivnosti konkretnog spoja, putu primjene, o uznapredovalosti stanja koje se tretira, te o stanju i povijesti bolesti bolesnika koji se tretira. Međutim, smatra da je unutar poznatog iskustva tehnike da se započne s razinama spoja koje su manje od potrebnih da bi se postigao željeni terapeutski učinak i da bi se postepeno povećalo doziranje sve dok se ne postigne željeni učinak. The actual dosage level of active ingredients in pharmaceutical compositions may vary to achieve a specific amount of active compound(s) effective to achieve a specific therapeutic response for a specific patient, composition, and route of administration. The selected dosage level depends on the activity of the specific compound, the route of administration, the advanced state of the condition being treated, and the condition and medical history of the patient being treated. However, it is believed to be within the known art to start with levels of the compound that are less than necessary to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved.

Općenito, oralno ili intravenski se bolesnom sisavcu primjenjuju doze od oko 0,1 do oko 1000 mg, oko 0,5 do oko 500 mg, oko 1 do oko 250 mg, oko 1,5 do oko 100, te poželjno oko 5 do oko 20 mg aktivnog spoja po kilogramu tjelesne težine. Po želji, učinkovita dnevna doza može se podijeliti u višestruke doze za potrebe primjene, npr. u dvije ili četiri odvojene doze dnevno. In general, doses of about 0.1 to about 1000 mg, about 0.5 to about 500 mg, about 1 to about 250 mg, about 1.5 to about 100, and preferably about 5 to about 20 mg of active compound per kilogram of body weight. If desired, the effective daily dose can be divided into multiple doses for administration purposes, for example, into two or four separate doses per day.

Ovaj izum prikazan je primjerima koji slijede. Primjer 1 prikazuje analizu visokog kapaciteta za detektiranje negativnih regulatora za LFA-1 vezanje za ICAM koji veće LFA-1. Primjer 2 odnosi se na vezujuću analizu za procjenu sposobnosti različitih spojeva u svezi s inhibiranjem LFA-1 vezanja za ICAM. Primjer 3 prikazuje sintezu negativnih regulatora. Primjer 4 prikazuje rezultate stanične analize korištenjem negativnih regulatora. This invention is illustrated by the following examples. Example 1 shows a high-throughput assay to detect negative regulators of LFA-1 binding to ICAMs that enhance LFA-1. Example 2 relates to a binding assay to assess the ability of various compounds to inhibit LFA-1 binding to ICAM. Example 3 shows the synthesis of negative regulators. Example 4 shows the results of a cellular assay using negative regulators.

Primjer 1 Example 1

Visokokapacitetni “screening” LFA-1/ICAM-1 vezujućih inhibitora High-throughput screening of LFA-1/ICAM-1 binding inhibitors

U nastojanju da se identificiraju inhibitori LFA-1/ICAM-1 vezanja, razrađena je visokokapacitetna “screening” analiza (HTS) da bi se ispitao veliki broj kemijskih spojeva iz vlastite biblioteke spojeva. In an effort to identify inhibitors of LFA-1/ICAM-1 binding, a high throughput screening assay (HTS) was developed to screen a large number of chemical compounds from our own compound library.

Prethodni eksperimenti su provedeni da bi se odredio linearni raspon LFA-1/ICAM-1 interakcije. Rekombinantni ICAM-1/IgGI protein (koji sadrži ICAM-1 pune duljine) priređen je kao kao što je opisano u američkim patentima br. 5,770,686, 5,837,478, i 5,869,262, a oba su ovdje uključena kao reference. Fuzijski protein je biotiniliran korištenjem pribora koji je dobiven od Pierce Chemical (Rockford, IL). Koncentracija biotiniliranog proteina (BioIgICAM-1) određena je mjerenjem apsorbancije na 280 nm, te su serijska razrjeđenja priređena da se dobije konačni koncentracijski raspon od 50 µg/ml do 0,008 µg/ml. Titriranje BioIgICAM-1 izvršeno je s proteinom koji je prvo razdijeljen na jažice ploče za analizu. Rekombinantni LFA-1 dodan je svakoj jažici u istoj koncentraciji i eksperiment (kao što je niže opisano) je proveden do završetka. Količina vezanja određena je za svaku jažicu, te je na temelju dijagrama rezultata odabrana jedna koncentracija BioIgICAM-1 za kasnije eksperimente. Na sličan način, titriran je LFA-1 korištenjem BioIgICAM-1 koncentracije koja je određena kao što je prije opisano. Previous experiments were performed to determine the linear range of the LFA-1/ICAM-1 interaction. Recombinant ICAM-1/IgGI protein (containing full-length ICAM-1) was prepared as described in US Pat. Nos. 5,770,686, 5,837,478, and 5,869,262, both of which are incorporated herein by reference. The fusion protein was biotinylated using a kit obtained from Pierce Chemical (Rockford, IL). The concentration of biotinylated protein (BioIgICAM-1) was determined by measuring absorbance at 280 nm, and serial dilutions were prepared to obtain a final concentration range of 50 µg/ml to 0.008 µg/ml. BioIgICAM-1 titration was performed with the protein that was first distributed to the wells of the analysis plate. Recombinant LFA-1 was added to each well at the same concentration and the experiment (as described below) was carried out to completion. The amount of binding was determined for each well, and one concentration of BioIgICAM-1 was selected for later experiments based on the result plot. Similarly, LFA-1 was titrated using BioIgICAM-1 concentration determined as previously described.

Prvog dana HTS postupka, tijelo za hvatanje tj. neblokirajuće anti-LFA-1 monoklonsko antitijelo (TS2/4.1; ATCC #HB244), razrijeđeno je puferom (50 mM natrijev karbonat/bikarbonat, 0,05% ProClin® 300, pH 9,6) do konačne koncentracije od 2 µg/ml. Immulon® 4 (Dynex Technologies, Chantilly, VA) jažice na ploči prevučene su sa 100 µl razrijeđenog antitijela po jažici, te je inkubacija provedena preko noći na 4°C. Drugog dana, ploče su zagrijana na sobnu temperaturu i isprane dva puta s puferom (fosfatna puferirana otopina bez kalcija i magnezija, CMF-PBS) s 0,05% Tween®-20). Svakoj jažici dodano je 200 µl blokirajuće otopine (5% želatine riblje kože u CMF-PBS s 0,05% ProClin® 300), te je provedeno blokirajuće inkubiranje na sobnoj temperaturi tijekom 30 min. Blokirajuća otopina je uklonjena sisanjem, te ploče nisu isprane. LFA-1 razrijeđen je na konačnu koncentraciju od 1 µg/ml u puferu za analizu (1% želatina riblje kože i 2 mM MgCl2 u CMF-PBS), te je u svaku jažicu dodano 100 µl. Inkubiranje je trajalo 1 sat i ploče su dva puta isprane puferom. On the first day of the HTS procedure, the capture body, i.e. non-blocking anti-LFA-1 monoclonal antibody (TS2/4.1; ATCC #HB244), was diluted with buffer (50 mM sodium carbonate/bicarbonate, 0.05% ProClin® 300, pH 9, 6) to a final concentration of 2 µg/ml. Immulon® 4 (Dynex Technologies, Chantilly, VA) plate wells were coated with 100 µl of diluted antibody per well, and incubation was performed overnight at 4°C. On the second day, the plates were warmed to room temperature and washed twice with buffer (calcium-magnesium-free phosphate-buffered saline, CMF-PBS) with 0.05% Tween®-20). 200 µl of blocking solution (5% fish skin gelatin in CMF-PBS with 0.05% ProClin® 300) was added to each well, and blocking incubation was performed at room temperature for 30 min. The blocking solution was removed by suction, and the plates were not washed. LFA-1 was diluted to a final concentration of 1 µg/ml in assay buffer (1% fish skin gelatin and 2 mM MgCl2 in CMF-PBS), and 100 µl was added to each well. Incubation lasted for 1 hour and the plates were washed twice with buffer.

Priređena je 2× “stock” otopina BioIgICAM-1 koja sadrži 0,1 µg/ml BioIgICAM-1 i 4 µM kristalnog ljubičastog (za koje je nađeno da je aktivator LFA—1/ICAM-1 vezanja) u puferu analize (EG&G Wallac, Gaithetsburg, MD). Alikvoti (50 µl) spojenih kemikalija (22 spoja/nakupina u 100% DMSO) iz kemijske biblioteke je dodano u jažice, a nakon toga je dodano 50 µl 2X “stock” BioIgICAM-1 da se dobije konačan volumen od 100 µl (sadrži 2% DMSO). Ploče su inkubirane jedan sat na sobnoj temperaturi, te isprane jednom puferom za ispiranje. Europij-obilježeni streptavidin (Eu-SA; #1244-360, EG&G Wallac) razrijeđen je 1:500 u puferu analize, 100 µl razrijeđenog Eu-SA dodano je u svaku jažicu, te su ploče inkubirane na sobnoj temperaturi jedan sat. A 2× stock solution of BioIgICAM-1 was prepared containing 0.1 µg/ml BioIgICAM-1 and 4 µM crystal violet (which was found to be an activator of LFA-1/ICAM-1 binding) in assay buffer (EG&G Wallac , Gaithetsburg, MD). Aliquots (50 µl) of pooled chemicals (22 compounds/pool in 100% DMSO) from the chemical library were added to the wells, followed by the addition of 50 µl 2X “stock” BioIgICAM-1 to give a final volume of 100 µl (containing 2 % DMSO). The plates were incubated for one hour at room temperature, and washed once with washing buffer. Europium-labeled streptavidin (Eu-SA; #1244-360, EG&G Wallac) was diluted 1:500 in assay buffer, 100 µl of diluted Eu-SA was added to each well, and the plates were incubated at room temperature for one hour.

Ploče su osam puta isprane puferom, u svaku jažicu dodano je 100 µl DELFIA® otopine za pojačavanje (EG&G Wallac) razrijeđene 1:2, te su ploče potresivane pet minuta pomoću Wallac mućkalice pri velikoj brzini. Ploče su očitane pomoću Wallac DELFIA® čitača fluorescencije (fluorimetar). Kontrole su se sastojale od pozitivnih i negativnih jažica i 50% vezujanih jažica koje su načinjene korištenjem blokirajućih antitijela, tj. anti-LFA-1 monoklonskih antitijela (TS 1/22.1, ATCC #HB202) ili anti-ICAM-1 monoklonskih antitijela. One nakupine (“pools”) koje su pokazale 50% ili veće inhibiranje LFA-1 vezanja za ICAM su identificirane i eksperiment je ponovljen korištenjem pojedinih kemikalija iz dotične nakupine. Određeni su inhibitori LFA-1/ICAM-1 vezanja, te je provedeno dodatno ispitivanje da se odredi ovisnost doze o inhibitorskoj aktivnosti. Izvršeno je daljnje ispitivanje odabranih spojeva tehnikama biokemijske i citološke analize. Plates were washed eight times with buffer, 100 µl DELFIA® amplification solution (EG&G Wallac) diluted 1:2 was added to each well, and plates were shaken for five minutes using a Wallac shaker at high speed. Plates were read using a Wallac DELFIA® fluorescence reader (fluorimeter). Controls consisted of positive and negative wells and 50% bound wells made using blocking antibodies, ie, anti-LFA-1 monoclonal antibody (TS 1/22.1, ATCC #HB202) or anti-ICAM-1 monoclonal antibody. Those pools that showed 50% or greater inhibition of LFA-1 binding to ICAM were identified and the experiment was repeated using individual chemicals from that pool. Inhibitors of LFA-1/ICAM-1 binding were determined, and an additional study was performed to determine the dose dependence of the inhibitory activity. Further testing of selected compounds was performed using biochemical and cytological analysis techniques.

Spojevi su grupirani sukladno zajedničkim strukturnim svojstvima, te je nađeno da sljedeći skup spojeva (navedeni niže) sadrži karakterističnu strukturu diaril sulfida. The compounds were grouped according to common structural properties, and the following set of compounds (listed below) were found to contain the characteristic diaryl sulfide structure.

3-Klor-4-(1-klor-nafthalen-2-ilsulfanil)-fenilamin 3-Chloro-4-(1-chloro-naphthalen-2-ylsulfanyl)-phenylamine

1-(3-Nitro-4-fenilsulfanil-fenil)-etanon 1-(3-Nitro-4-phenylsulfanyl-phenyl)-ethanone

1-(3-Nitro-4-fenilsulfanil-fenil)-etanon oksim 1-(3-Nitro-4-phenylsulfanyl-phenyl)-ethanone oxime

5-Trifluormetil-2-fenilsulfanil-benzonitril 5-Trifluoromethyl-2-phenylsulfanyl-benzonitrile

1-(3,5-diklorfenil)-3-fenilsulfanil-pirolidin-2,5-dion 1-(3,5-dichlorophenyl)-3-phenylsulfanyl-pyrrolidine-2,5-dione

Bis-2,4,6-Trinitrofenil-sulfid Bis-2,4,6-Trinitrophenyl-sulphide

2-Metil-1-(2-o-tolilsulfanil-fenil)-1H-pirol 2-Methyl-1-(2-o-tolylsulfanyl-phenyl)-1H-pyrrole

3-[2-(4-Klor-2-nitro-fenilsulfanil)-fenilamino-3H-izobenzofuran-1-on 3-[2-(4-Chloro-2-nitro-phenylsulfanyl)-phenylamino-3H-isobenzofuran-1-one

4-(Benzotiazol-2-ilsulfanil)-3-klor-fenilamin 4-(Benzothiazol-2-ylsulfanyl)-3-chloro-phenylamine

2-Nitro-4-klorfenil-(2'-aminofenil)-sulfid 2-Nitro-4-chlorophenyl-(2'-aminophenyl)-sulfide

6-Amino-2-klorfenil-(4'-metilfenil)-sulfid 6-Amino-2-chlorophenyl-(4'-methylphenyl)-sulfide

4-Nitrofenil-(2'-klorfenil)-sulfid 4-Nitrophenyl-(2'-chlorophenyl)sulfide

2, 4-Dinitrofenil-(4'-klorfenil)-sulfid 2, 4-Dinitrophenyl-(4'-chlorophenyl)-sulfide

4-Aminofenil-(2'-klorfenil)-sulf de 4-Aminophenyl-(2'-chlorophenyl)-sulf de

2, 4-Diaminofenil-(4'-izopropilfenil)-sulfid 2, 4-Diaminophenyl-(4'-isopropylphenyl)-sulfide

U nastojanju da se optimizira negativni regulacijski kapacitet identificiranih spojeva, načinjeni su različiti derivati diaril sulfida kao što je opisano u primjeru 3. Dodatni derivati diaril sulfida opisani su u pratećoj patentnoj priujavi naslovljenoj “Protivupalni i imuno-supresivni spojevi koji inhibiraju staničnu adheziju” koja je podnesena 2. travnja 1999, zastupnički broj 6446.US.Z3, serijski broj 09/286 645, koja je ovdje u cijelosti uključena kao referenca. In an effort to optimize the negative regulatory capacity of the identified compounds, various diaryl sulfide derivatives were made as described in Example 3. Additional diaryl sulfide derivatives are described in a companion patent application entitled "Anti-inflammatory and immuno-suppressive compounds that inhibit cell adhesion" which filed Apr. 2, 1999, Rep. No. 6446.US.Z3, Serial No. 09/286,645, which is incorporated herein by reference in its entirety.

Primjer 2 Example 2

Vezujuće analize Binding analyses

A. Analiza ICAM-1/LFA-1 biokemijske interakcije A. Analysis of ICAM-1/LFA-1 biochemical interaction

Spojevi koji antagoniziraju interakciju između ICAM-1 i LFA-1 mogu biti identificirani i njihova aktivnost izmjerena korištenjem biokemijskih i staničnih analiza. Primarna biokemijska analiza mjeri sposobnost konkretnog spoja da blokira interakciju između LFA-1 i njegova adhezijskog partnera ICAM-l, kao što je niže objašnjeno. Compounds that antagonize the interaction between ICAM-1 and LFA-1 can be identified and their activity measured using biochemical and cellular assays. A primary biochemical assay measures the ability of a particular compound to block the interaction between LFA-1 and its adhesion partner ICAM-1, as discussed below.

U ovoj biokemijskoj analizi, 100 µl anti-LFA-1 antitijela u svezi 5 µg/ml u Dulbecco-voj fosfatno puferiranoj otopini soli (D-PBS) korišteno je za prevlačenje jažica mikrotitarske ploče s 96 jažica preko noći na 4°C. Jažice su dva puta isprane puferom za ispiranje (CMF-PBS, 0,05% Tween® 20) i blokirane dodatkom 200 µl D-PBS koji sadrži 5% želatinu riblje kože. U svaku jažicu dodan je rekombinantni LFA-1 (100 µl 0,7 µg/ml) u D-PBS. Inkubiranje je nastavljeno jedan sat na sobnoj temperaturi i jažice su isprane dva puta puferom za ispiranje. Serijska razrjeđenja spojeva koji su ispitani kao LFA-1/ICAM-I negativni regulatori priređena su kao 10 mM “stock” otopine u DMSO, razrijeđena u D-PBS, 2 mM MgCl2, 1% želatine riblje kože i 50 µl svakog razrjeđenja dodano je u svaku jažicu, nakon čega je slijedio dodatak 50 µl 0,8 pg/ml BioIgICAM-1 svakoj jažici, te su ploče inkubirane na sobnoj temperaturi jedan sat. Jažice su zatim isprane dva puta puferom za ispiranje i jažicama je dodano 100 µl Eu-SA (EG&G Wallac) razrijeđeno je 1:100 u Delfia® puferu analize (EG&G Wallac). Inkubiranje je provedeno jedan sat na sobnoj temperaturi. Jažice su isprane osam puta puferom za ispiranje i u svaku jažicu dodano je 100 µ1 otopine za pojačavanje (EG&G Wallac). Inkubiranjem je nastavljeno pet minuta uz konstantno miješanje. Vremenski razlučena fluorimetrijska mjerenja izvršena su korištenjem Victor 1420 Multilabel Counter (EG&G Wallac) brojača i postotak inhibiranja svakog potencijalnog spoja izračunat je pomoću sljedeće jednadžbe: In this biochemical assay, 100 µl of anti-LFA-1 antibody at 5 µg/ml in Dulbecco's phosphate-buffered saline (D-PBS) was used to coat the wells of a 96-well microtiter plate overnight at 4°C. The wells were washed twice with wash buffer (CMF-PBS, 0.05% Tween® 20) and blocked by the addition of 200 µl D-PBS containing 5% fish skin gelatin. Recombinant LFA-1 (100 µl 0.7 µg/ml) in D-PBS was added to each well. Incubation was continued for one hour at room temperature and the wells were washed twice with wash buffer. Serial dilutions of compounds tested as LFA-1/ICAM-I negative regulators were prepared as 10 mM stock solutions in DMSO, diluted in D-PBS, 2 mM MgCl2, 1% fish skin gelatin, and 50 µl of each dilution was added into each well, followed by the addition of 50 µl of 0.8 pg/ml BioIgICAM-1 to each well, and the plates were incubated at room temperature for one hour. The wells were then washed twice with wash buffer and 100 µl Eu-SA (EG&G Wallac) diluted 1:100 in Delfia® assay buffer (EG&G Wallac) was added to the wells. Incubation was carried out for one hour at room temperature. Wells were washed eight times with wash buffer and 100 µl of amplification solution (EG&G Wallac) was added to each well. Incubation was continued for five minutes with constant mixing. Time-resolved fluorimetric measurements were performed using a Victor 1420 Multilabel Counter (EG&G Wallac) and the percent inhibition of each potential compound was calculated using the following equation:

[image] [image]

gdje se bkg odnosi na jažice koje nisu prevučene s anti-FA-1 antitijelom. where bkg refers to wells not coated with anti-FA-1 antibody.

B. Analiza ICAM-1/JY-8 stanične adhezije B. ICAM-1/JY-8 cell adhesion assay

Biološki relevantna aktivnost spojeva ovog izuma potvrđena je analizom stanične adhezije kojom se mjeri sposobnost spojeva da blokiraju adherenciju JY-8 stanica (humana EBV-transformirana B stanična linija s ekspresijom LFA-1 na njegovoj površini) za imobilizirani ICAM-1, na sljedeći način. Ova analiza može se provesti sa i bez dodanog IL-8. Za IL8 stimuliranje standardnih JY-8 stanica dodano je 30 ng/ml IL-8 za 30 minutno inkubiranje na 37°C. The biologically relevant activity of the compounds of this invention was confirmed by a cell adhesion assay measuring the ability of the compounds to block the adherence of JY-8 cells (a human EBV-transformed B cell line expressing LFA-1 on its surface) to immobilized ICAM-1, as follows. This assay can be performed with and without added IL-8. For IL8 stimulation of standard JY-8 cells, 30 ng/ml IL-8 was added for 30 minutes incubation at 37°C.

Za mjerenje aktivnosti inhibiranja u analizi stanične adhezije, ploče s 96 jažica prevučene su sa 70 µl rekombinantnog ICAM-1/Ig s koncentracijom 5 µg/ml u CMF-PBS preko noći 4°C. Jažice su isprane dva puta s D-PBS i blokirane dodatkom 200 µl D-PBS, 5% želatine riblje kože inkubiranjem jedan sat na sobnoj temperaturi. Fluorescentno obilježene JY-8 stanice (50 µl za 2×106 stanica/ml u RPMI-1640/1% fetalnog volovskog seruma (FBS)) dodane su u jažice. Za fluorescentno obilježavanje JY-8 stanica, 5×106 stanica isprano je jednom u RPMI 1640, resuspendirano u 1 ml RPMI-1640 koji sadrži 2 uM kalcein AM (Molecular Probes, OR), inkubirano na 37°C tijekom 30 minuta i isprano je jednom s RPMI-1640/1% FBS. Razrjeđenja spojeva koji se ispituju na LFA-1/ICAM-3 antagonističku aktivnost načinjena su u adhezijskom puferu iz 10 mM “stock” otopina u DMSO i 50 µl alikvoti dodani su u duplikatne jažice. Mikrotitarske ploče inkubirane su 45 minuta na sobnoj temperaturi i jažice su isprane blago jednom s RPMI-1640/1% FBS. Intenzitet fluorescencije izmjeren je u čitaču fluorescencije za ploče s valnom duljinom pobude na 495 nm i valnom duljinom emisije na 530 nm. Postotak inhibiranja potencijalnog spoja za danu koncentraciju izračunat je prema sljedećoj jednadžbi: To measure the inhibitory activity in the cell adhesion assay, 96-well plates were coated with 70 µl recombinant ICAM-1/Ig at a concentration of 5 µg/ml in CMF-PBS overnight at 4°C. The wells were washed twice with D-PBS and blocked by adding 200 µl of D-PBS, 5% fish skin gelatin by incubating for one hour at room temperature. Fluorescently labeled JY-8 cells (50 µl for 2×106 cells/ml in RPMI-1640/1% fetal bovine serum (FBS)) were added to the wells. For fluorescent labeling of JY-8 cells, 5×106 cells were washed once in RPMI 1640, resuspended in 1 ml of RPMI-1640 containing 2 µM calcein AM (Molecular Probes, OR), incubated at 37°C for 30 min, and washed once with RPMI-1640/1% FBS. Dilutions of compounds to be tested for LFA-1/ICAM-3 antagonistic activity were made in adhesion buffer from 10 mM "stock" solutions in DMSO and 50 µl aliquots were added to duplicate wells. Microtiter plates were incubated for 45 min at room temperature and the wells were washed gently once with RPMI-1640/1% FBS. Fluorescence intensity was measured in a plate fluorescence reader with an excitation wavelength of 495 nm and an emission wavelength of 530 nm. The percentage of inhibition of a potential compound for a given concentration was calculated according to the following equation:

[image] [image]

C. Analiza ICAM-31/Y-8 stanične adhezije C. ICAM-31/Y-8 cell adhesion assay

Može se pokazati da spojevi ovog izuma djeluju putem interakcije s integrinom LFA-l, specifično vezanjem αL I domene za koju se zna da je kritična za adheziju LFA-1 s cijelim nizom molekula stanične adhezije. Kao takvi, očekuje se da bi ti spojevi mogli blokirati interakciju LFA-1 s drugim CAM-ima, te je ovo inhibiranje demonstrirano LFA-1 vezanjem za ICAM-3. Spojevi ovog izuma procijenjeni su glede sposobnosti blokiranja dhezije JY-8 stanica za imobilizirani ICAM-3, na sljedeći način. The compounds of the present invention can be shown to act through interaction with the LFA-1 integrin, specifically by binding the αL I domain known to be critical for the adhesion of LFA-1 to a variety of cell adhesion molecules. As such, it is expected that these compounds could block the interaction of LFA-1 with other CAMs, and this inhibition was demonstrated by LFA-1 binding to ICAM-3. The compounds of this invention were evaluated for their ability to block the adhesion of JY-8 cells to immobilized ICAM-3, as follows.

Za mjerenje aktivnosti inhibiranja u analizi stanične adhezije, mikrotitarske ploče s 96 jažica prevučene su s 50 µl rekombinantnog ICAM-3/Ig s koncentracijom 10 µg/ml u CMF-PBS preko noći na 4°C. Jažice su isprane dva puta s D-PBS, blokirane dodatkom 100 µl D-PBS, 1% volovskog serum albumina (BSA) inkubiranjem jedan sat na sobnoj temperaturi, te su isprane jednom s RPM1-1640/5% toplinski inaktiviranim FBS (adhezijski pufer). Razrjeđenja spojeva koji se ispituju na LFA-1/ICAM-3 antagonističku aktivnost načinjena su u adhezijskom puferu iz 10 mM “stock” otopina u DMSO i 100 µl alikvoti dodani su u duplikatne jažice. Zatim su JY-8 stanice (100 µl pri 0,75×106 stanica/ml u adhezijskom puferu) dodane jažicama (sa i bez 30 ng/ml IL-8). Mikrotitarske ploče inkubirane su 30 minuta na sobnoj temperaturi, adherentne stanice fiksirane su s 50 µ1 14% glutaraldehid/D-PBS i provedeno je inkubiranje daljnjih 90 minuta. Jažice su isprane blago s dH2O i dodano je 50 µl dH2O, a nakon toga 50 µl 1% kristalno ljubičastog. Nakon pet minuta, ploče su isprane dva puta s dH2O i u svaku jažicu je dodano 225 µl etanola (EtOH) da se ekstrahira kristalno ljubičasto iz stanica. Apsorbancija je izmjerena na 570 nm u ELISA čitaču ploča. Postotak inhibiranja potencijalnog spoja izračunat je prema sljedećoj jednadžbi: To measure the inhibitory activity in the cell adhesion assay, 96-well microtiter plates were coated with 50 µl recombinant ICAM-3/Ig at a concentration of 10 µg/ml in CMF-PBS overnight at 4°C. Wells were washed twice with D-PBS, blocked with 100 µl of D-PBS, 1% bovine serum albumin (BSA) by incubating for one hour at room temperature, and washed once with RPM1-1640/5% heat-inactivated FBS (adhesion buffer ). Dilutions of compounds to be tested for LFA-1/ICAM-3 antagonistic activity were made in adhesion buffer from 10 mM "stock" solutions in DMSO and 100 µl aliquots were added to duplicate wells. Then, JY-8 cells (100 µl at 0.75×10 6 cells/ml in adhesion buffer) were added to the wells (with and without 30 ng/ml IL-8). Microtiter plates were incubated for 30 minutes at room temperature, adherent cells were fixed with 50 µl of 14% glutaraldehyde/D-PBS and incubation was carried out for a further 90 minutes. The wells were washed gently with dH2O and 50 µl of dH2O was added followed by 50 µl of 1% crystal violet. After five minutes, the plates were washed twice with dH2O and 225 µl ethanol (EtOH) was added to each well to extract the crystal violet from the cells. Absorbance was measured at 570 nm in an ELISA plate reader. The percentage of potential compound inhibition was calculated according to the following equation:

[image] [image]

Primjer 3 Example 3

Sinteza negativnih regulatora Synthesis of negative regulators

U nastavku je opisana sinteza različitih diaril sulfidnih spojeva sukladno ovom izumu. The synthesis of various diaryl sulfide compounds according to this invention is described below.

Opći postupci i polazne tvari General procedures and starting materials

Općenito, otapala su nabavljena bezvodna te nisu pročišćavana ili sušena, a plazne tvari su najbolje koje se mogu nabaviti. Tankoslojna kromatografija (TLC) načinjena je korištenjem staklenih ili aluminijskih ploča E. Merck, silika-gel 60 F254, 0,25 mm, ili Analtech silika-gel ploča (250 mikrona silika-gela). Vizualiziranje je izvršeno pomoću UV svjetla. Navedene vrijednosti Rf označuju jednu detektiranu mrlju. Spektri nuklearne magnetske rezonancije (NMR) načinjeni su na Bruker DPX 300 ili Varian 300 Gemini 2000 spektrometrima. Kemijski pomaci navedeni su u dijelovima na milijun (ppm) u odnosu na tetrametilsilan (TMS). Konstante vezanja su u Hz. U drugim slučajevima, NMR spektri su zabilježeni pomoću Unity XL-200 spektrometra. Maseni spektri su snimljeni na VG 70 SEG instrumentu na University of Washington, Department of Medicinal Chemistry, Mass Spectrometry Facility (visoko razlučivanje) ili na Finnigan Mat TSQ 70 spektrometru (nisko razlučivanje). Navedeni su samo molekularni ioni. Elementarna analiza je izvršena pomoću Quantitative Technologies, Inc. In general, the solvents obtained are anhydrous and not purified or dried, and plasma substances are the best that can be obtained. Thin layer chromatography (TLC) was performed using E. Merck glass or aluminum plates, silica gel 60 F254, 0.25 mm, or Analtech silica gel plates (250 micron silica gel). Visualization was performed using UV light. The specified Rf values indicate one detected spot. Nuclear magnetic resonance (NMR) spectra were taken on Bruker DPX 300 or Varian 300 Gemini 2000 spectrometers. Chemical shifts are given in parts per million (ppm) relative to tetramethylsilane (TMS). Binding constants are in Hz. In other cases, NMR spectra were recorded using a Unity XL-200 spectrometer. Mass spectra were recorded on a VG 70 SEG instrument at the University of Washington, Department of Medicinal Chemistry, Mass Spectrometry Facility (high resolution) or on a Finnigan Mat TSQ 70 spectrometer (low resolution). Only molecular ions are listed. Elemental analysis was performed using Quantitative Technologies, Inc.

A. Opća sintetska metoda A: Aminodiarilsulfidi A. General synthetic method A: Aminodiarylsulfides

B. 1. Opći opis sintetske metode A B. 1. General description of the synthetic method A

Jedan molarni ekvivalent (ekv.) željenog tiolfenola i 1 molarni ekvivalent odgovarajućeg nitroarilnog spoja stavljeni su u suhu tikvicu pod dušikom i otopljeni u suhom acetonu. Dodan je jedan i pol molarni ekvivalent bezvodnog kalijeva karbonata (K2CO3) i smjesa je energično miješana preko noći. Reakcijska smjesa je zatim razrijeđena eterom, isprana zasićenom otopinom NaHCO3, 3% otopinom NaHSO4 i zasićenom otopinom NaCl. Organski slojevi su osušeni iznad Na2SO4 i filtrirani. Dodan je heptan i otopina je koncentrirana vrijanjem sve dok nije uklonjena većina etera. Nakon hlađenja, kristalizirao je nitrodiaril sulfid. Produkt je sakupljen filtriranjem, ispran pentanom i osušen u vakuumu. One molar equivalent (eq) of the desired thiolphenol and 1 molar equivalent of the corresponding nitroaryl compound were placed in a dry flask under nitrogen and dissolved in dry acetone. One and a half molar equivalent of anhydrous potassium carbonate (K2CO3) was added and the mixture was stirred vigorously overnight. The reaction mixture was then diluted with ether, washed with saturated NaHCO3 solution, 3% NaHSO4 solution and saturated NaCl solution. The organic layers were dried over Na2SO4 and filtered. Heptane was added and the solution was concentrated by boiling until most of the ether was removed. After cooling, nitrodiaryl sulfide crystallized. The product was collected by filtration, washed with pentane and dried in vacuo.

Jedan molarni ekvivalent diaril sulfida i 5 molarnih ekvivalenata kositrenog klorida dihidrata otopljeno je u etanolu (10 do 30 volumena). Smjesa je zagrijana na 60°C u uljnoj kupelji i dodana je koncentrirana HCl (10 do 30 volumena). Nakon tri sata, reakcijska smjesa je ostavljena da se ohladi i dodano je 20 do 60 volumnih dijelova leda. Smjesa je neutralizirana na pH 10-12 dodatkom 5N otopine NaOH. Smjesa je dva puta ekstrahirana eterom i sjedinjeni eterski ekstrakti isprani su zasićenom otopinom NaHCO3 i zasićenom otopinom NaCl. Organski sloj je osušen iznad Na2SO4, filtriran i otapalo je uklonjeno rptacijskim uparavanjem. Dobivena krutina ili ulje je sakupljeno eterom, te je dokapavanjem dodano približno 5 molarnih ekvivalenata 1 N otopine HCl u eteru. Dobivena krutina je sakupljena filtriranjem, isprana eterom, pa osušena u vakuumu. One molar equivalent of diaryl sulfide and 5 molar equivalents of stannous chloride dihydrate were dissolved in ethanol (10 to 30 volumes). The mixture was heated to 60°C in an oil bath and concentrated HCl (10 to 30 volumes) was added. After three hours, the reaction mixture was allowed to cool and 20 to 60 parts by volume of ice was added. The mixture was neutralized to pH 10-12 by the addition of 5N NaOH solution. The mixture was extracted twice with ether and the combined ether extracts were washed with saturated NaHCO3 solution and saturated NaCl solution. The organic layer was dried over Na 2 SO 4 , filtered and the solvent was removed by evaporation. The resulting solid or oil was collected with ether, and approximately 5 molar equivalents of a 1 N solution of HCl in ether was added dropwise. The resulting solid was collected by filtration, washed with ether, and dried in vacuo.

Opća metoda sinteze A je shematski prikazana sljedećim dijagramom, gdje je X halogen, primjerice klor. The general method of synthesis of A is shown schematically in the following diagram, where X is a halogen, for example chlorine.

[image] [image]

2. Specifični primjer opće sintetske metode A 2. Specific example of general synthetic method A

Korištena je opća sintetska metoda A kao što je prije opisano, s izuzetkom što su korištene granule kositra umjesto poželjnog kositrenog klorida dihidrata, kao što je prikazano prije shematski. General synthetic method A was used as previously described, with the exception that tin granules were used instead of the preferred stannous chloride dihydrate, as shown schematically above.

2,4-Diklorfenil-(2'-klor-4'-nitrofenil)-sulfid 2,4-Dichlorophenyl-(2'-chloro-4'-nitrophenyl)-sulfide

1,54 g (8,60 mmol) 2,4-diklortiofenola i 1,65 g (1 ekv., 8,60 mmol) 3,4-diklornitrobenzena stavljeno je u suhu tikvicu u atmosferi dušika i otopljeno u suhom acetonu. Dodano je 1,78 g (1,5 ekv., 12,9 mmol) bezvodnog kalijeva karbonata (K2CO3) i magnetski je miješano 20 sati. Reakcijska smjesa je razrijeđena sa 150 ml etera i isprana zasićenom otopinom NaHCO3 (2×75 ml), 3% otopinom NaHSO3 (2×75 ml) i zasićenom otopinom NaCl (2×75 ml). Organska faza je osušena iznad bezvodnog NaHSO3, te filtrirana. Dodano je 50 ml heptana, te je otopina koncentrirana na približno 50 ml vrenjem u parnoj kupelji. Nakon hlađenja dobiveni su visokokvalitetni rkistali. Sakupljeni su filtriranjem, isprani s tri obroka pentana, te osušeni u vakuumu (70°C, 0,05 mm Hg, 2 sata). Dobiveno je 1,43 g (50% prinos) žutih kristala, talište 145-146°C, Rf 0,37 (etilacetat [EtAc]/Heptan, 1:20). 1H-NMR (CDCl3) 6.71 (d, J=8.9, 1H), 7.38 (d od d, J1=2.2, J2=8.2, 1H), 7.59 (d, J=8.2, 1H), 7.63 (d, J=2.2, 1H), 7.94 (d od d, J1=2.2, J2=8.9, 1H), 8.26 (d;1=2.5, 1H). MS (EI) m/z 333 (M+, 98). Analitički izračunato za C12H6Cl3NO2S: C, 43.08; H, 1.81; N, 4.19. Nađeno: C, 43.06; H, 1.77; N, 4.02. 1.54 g (8.60 mmol) of 2,4-dichlorothiophenol and 1.65 g (1 eq., 8.60 mmol) of 3,4-dichloronitrobenzene were placed in a dry flask under nitrogen and dissolved in dry acetone. 1.78 g (1.5 eq., 12.9 mmol) of anhydrous potassium carbonate (K 2 CO 3 ) was added and magnetically stirred for 20 hours. The reaction mixture was diluted with 150 ml of ether and washed with saturated NaHCO3 solution (2×75 ml), 3% NaHSO3 solution (2×75 ml) and saturated NaCl solution (2×75 ml). The organic phase was dried over anhydrous NaHSO3 and filtered. 50 ml of heptane was added, and the solution was concentrated to approximately 50 ml by boiling in a steam bath. After cooling, high-quality crystals were obtained. They were collected by filtration, washed with three portions of pentane, and dried in vacuum (70°C, 0.05 mm Hg, 2 hours). 1.43 g (50% yield) of yellow crystals were obtained, mp 145-146°C, Rf 0.37 (ethyl acetate [EtAc]/Heptane, 1:20). 1H-NMR (CDCl3) 6.71 (d, J=8.9, 1H), 7.38 (d from d, J1=2.2, J2=8.2, 1H), 7.59 (d, J=8.2, 1H), 7.63 (d, J =2.2, 1H), 7.94 (d of d, J1=2.2, J2=8.9, 1H), 8.26 (d;1=2.5, 1H). MS (EI) m/z 333 (M+, 98). Analytical calculated for C12H6Cl3NO2S: C, 43.08; H, 1.81; N, 4.19. Found: C, 43.06; H, 1.77; N, 4.02.

3-Klor-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 3-Chloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride

0,680 g (2,03 mmol) 2,4-Diklorfenil-(2'-klor-4'-nitrofenil)-sulfida i 0,844 g (3,5 ekv., 7,11 mmol) kositrenih granula razmuljeno je u 20 ml koncentrirane HCl. Uz snažno miješanje, grijano je pod refluksom dva dana. Smjesa je ostavljena da se ohladi na sobnu temperaturu i razrijeđena je s 50 ml leda. Uz miješanje, smjesa je neutralizirana na pH 10 dodatkom približno 80 ml 5 N otopine NaOH. Izvršeno je ekstrahiranje eterom (2×100 ml). Sjedinjeni organski slojevi isprani su sa 100 ml zasićene otopine NaHCO3 i 2×100 ml zasićene otopine NaCI. To je osušeno iznad Na2SO4 i filtrirano. Izvršeno je koncentriranje na rotavaporu da se dobije smeđe ulje. Ulje je pokupljeno s 50 ml etera, dodano je 4 ml 1 N otopine HCI u eteru, dokapavanjem, da se dobije bijela krutina. Krutina je sakupljena filtriranjem, isprana s nekoliko dijelova etera, osušena (90°C, 3 sata, 0,05 mm Hg). Dobiveno je 0,548 g (79% prinos) bijele krutine, talište 183-185°C (raspad), Rf 0,33 (EtAc/Heptan 1:2 w/ 1% TEA). 1H-NMR (DMSO-d6) 6.55 (d, J=8.6, 1H), 6.66 (d od d, J1=2.5, J2=8.6, 1H), 6.89 (br s, 3H), 6.89 (d, J=2.5, 1H), 7.29 (d, J=2.2, 1H), 7.32 (d, J=2.2, 1H), 7.62 (d, J=2.2, 1H). MS (EI) m/z 303 M+ [100]. Analitički izračunato za C12H8Cl3NS·HCl: C, 42.26; H, 2.66; N, 4.11. Nađeno: C, 42.39; H, 2.57; N, 4.14. 0.680 g (2.03 mmol) of 2,4-Dichlorophenyl-(2'-chloro-4'-nitrophenyl)-sulfide and 0.844 g (3.5 eq., 7.11 mmol) of tin granules were dissolved in 20 ml of concentrated HCl. With vigorous stirring, it was heated under reflux for two days. The mixture was allowed to cool to room temperature and was diluted with 50 ml of ice. With stirring, the mixture was neutralized to pH 10 by the addition of approximately 80 ml of 5 N NaOH solution. Extraction was performed with ether (2×100 ml). The combined organic layers were washed with 100 ml of saturated NaHCO3 solution and 2×100 ml of saturated NaCl solution. This was dried over Na2SO4 and filtered. Concentration was carried out on a rotavapor to give a brown oil. The oil was taken up with 50 ml of ether, 4 ml of 1 N HCl in ether was added dropwise to give a white solid. The solid was collected by filtration, washed with several portions of ether, dried (90°C, 3 hours, 0.05 mm Hg). 0.548 g (79% yield) of a white solid was obtained, mp 183-185°C (dec), Rf 0.33 (EtAc/Heptane 1:2 w/ 1% TEA). 1H-NMR (DMSO-d6) 6.55 (d, J=8.6, 1H), 6.66 (d of d, J1=2.5, J2=8.6, 1H), 6.89 (br s, 3H), 6.89 (d, J= 2.5, 1H), 7.29 (d, J=2.2, 1H), 7.32 (d, J=2.2, 1H), 7.62 (d, J=2.2, 1H). MS (EI) m/z 303 M+ [100]. Analytical calculated for C12H8Cl3NS·HCl: C, 42.26; H, 2.66; N, 4.11. Found: C, 42.39; H, 2.57; N, 4.14.

3. Dodatni primjeri spojeva koji su priređeni općom sintetskom metodom A 3. Additional examples of compounds prepared by general synthetic method A

Sljedeći spojevi priređeni su iz komercijalno raspoloživih reagnesa korištenjem opće sintetske metode A. Načinjeni su polazni tiolni i nitroarilni spojevi, zajedno sintermedijerima (tiol+nitroaril-intermedijer). The following compounds were prepared from commercially available reagents using the general synthetic method A. The initial thiol and nitroaryl compounds were prepared together with intermediates (thiol+nitroaryl-intermediate).

3-Klor-4-(2-klorfenilsulfanil}-fenilamin hidroklorid 3-Chloro-4-(2-chlorophenylsulfanyl}-phenylamine hydrochloride).

2-klortiofenol + 3,4-diklornitrobenzen - 4-nitro-2-klorfenil-(2'-klorfenil)-sulfid 2-chlorothiophenol + 3,4-dichloronitrobenzene - 4-nitro-2-chlorophenyl-(2'-chlorophenyl)-sulfide

Siva krutina, talište 197-198°C (raspad), Rf 0,16 (EtAc/Heptan 1:4 w/ 1% TEA). 1H-NMR (DMSO-d6) 6.61 (d od d, J1=1.9, J2=7.3, 1H), 6.70 (d od d, J1=2.4, J2=8.4, 1 H), 6.94 (d, J=2.5, 1H), 7.18 (m, 2H), 7.28 (d, J=8.9, 1H), 7.45 (d od d, J1=1.6, J2=7.6, 1H), 7.78 (br s, 3H). MS (EI) m/z 269 M+ [100]. Analitički izračunato za C12H10Cl3NS·HCl: C, 47.22; H, 3.30; N, 4.59. Nađeno: C, 47.34; H, 3.15; N, 4.45. Gray solid, mp 197-198°C (dec), Rf 0.16 (EtAc/Heptane 1:4 w/ 1% TEA). 1H-NMR (DMSO-d6) 6.61 (d from d, J1=1.9, J2=7.3, 1H), 6.70 (d from d, J1=2.4, J2=8.4, 1H), 6.94 (d, J=2.5 , 1H), 7.18 (m, 2H), 7.28 (d, J=8.9, 1H), 7.45 (d of d, J1=1.6, J2=7.6, 1H), 7.78 (br s, 3H). MS (EI) m/z 269 M+ [100]. Analytical calculated for C12H10Cl3NS·HCl: C, 47.22; H, 3.30; N, 4.59. Found: C, 47.34; H, 3.15; N, 4.45.

3-Klor-4-(2-naftilsulfanil)-fenilamin hidroklorid 3-Chloro-4-(2-naphthylsulfanyl)-phenylamine hydrochloride

2-naftalentiol + 3,4-diklornitrobenzen - 4-nitro-2-klorfenil-2-naftilsulfid 2-naphthalenethiol + 3,4-dichloronitrobenzene - 4-nitro-2-chlorophenyl-2-naphthylsulfide

Prljavobijela krutina, talište 188°C (raspad), Rf 0,16 (EtAc/heptan 1:4 s 1% TEA). 1H-NMR (DMSO-d6) 6.08 (br s, 3H), 6.60 (d od d, J1=2.4, J2=8.4, 1H), 6.90 (d, J=2.5, 1H), 7.20 (d od d, J1=1.9, J2=8.6, 1H), 7.30 (d, J=8.6, 1H), 7.45 (m, 2H), 7.53 (s, 1H), 7.76 (m, 1H), 7.83 (m, 2H). MS (EI) m/z 285 M+ [100]. Analitički izračunato za C16H12ClNS·HCl: C, 59.64; H, 4.07; N, 4.35. Nađeno: C, 59.59; H, 4.07; N, 4.09. Off-white solid, mp 188°C (dec), Rf 0.16 (EtAc/heptane 1:4 with 1% TEA). 1H-NMR (DMSO-d6) 6.08 (br s, 3H), 6.60 (d from d, J1=2.4, J2=8.4, 1H), 6.90 (d, J=2.5, 1H), 7.20 (d from d, J1=1.9, J2=8.6, 1H), 7.30 (d, J=8.6, 1H), 7.45 (m, 2H), 7.53 (s, 1H), 7.76 (m, 1H), 7.83 (m, 2H). MS (EI) m/z 285 M+ [100]. Analytical calculated for C16H12ClNS·HCl: C, 59.64; H, 4.07; N, 4.35. Found: C, 59.59; H, 4.07; N, 4.09.

3-Klor-4-(2,3-diklorfenilsulfanil)-fenilamin hidroklorid 3-Chloro-4-(2,3-dichlorophenylsulfanyl)-phenylamine hydrochloride

2,3-diklortiofenol +3,4-diklornitrobenzen- 4-nitro-klorfenil-(2',3'-diklorfenil)-sulfid 2,3-dichlorothiophenol +3,4-dichloronitrobenzene-4-nitro-chlorophenyl-(2',3'-dichlorophenyl)-sulfide

Prljavobijela krutina, talište 207-209°C (raspad), Rf 0,33 (EtAc/heptan 1:2 s 1% TEA). 1H-NMR (DMSO-d6) 6.45 (d od d, J1=1.4, J2=8.1, 1H), 6.62 (d od d, J1=2.2, J2=8.6, 1 H), 6.79 (br s, 3H), 6.86 (d, J=2.5, 1 H), 7.21 (t, J=7.9, 1 H), 7.33 (d, J=8.2, 1H), 7.38 (d od d, J1=1.4, J2=8.1, 1H). MS (EI) m/z 303 (M+, 93). Analitički izračunato za C12H8Cl3NS·HCl: C, 42.26; H, 2.66; N, 4.11. Nađeno: C, 42.49; H, 2.56; N, 4.10. Off-white solid, mp 207-209°C (dec), Rf 0.33 (EtAc/heptane 1:2 with 1% TEA). 1H-NMR (DMSO-d6) 6.45 (d from d, J1=1.4, J2=8.1, 1H), 6.62 (d from d, J1=2.2, J2=8.6, 1H), 6.79 (br s, 3H) , 6.86 (d, J=2.5, 1 H), 7.21 (t, J=7.9, 1 H), 7.33 (d, J=8.2, 1H), 7.38 (d from d, J1=1.4, J2=8.1, 1H). MS (EI) m/z 303 (M+, 93). Analytical calculated for C12H8Cl3NS·HCl: C, 42.26; H, 2.66; N, 4.11. Found: C, 42.49; H, 2.56; N, 4.10.

3-Klor-4-(2,4,5-triklorfenilsulfanil)-fenilamin hidroklorid 3-Chloro-4-(2,4,5-trichlorophenylsulfanyl)-phenylamine hydrochloride

2,3,4-triklortiofenol + 3,4-diklornitrobenzen-4-nitro-2-klorfenil(2',4',5'-triklorfenil)-sulfid 2,3,4-trichlorothiophenol + 3,4-dichloronitrobenzene-4-nitro-2-chlorophenyl(2',4',5'-trichlorophenyl)-sulfide

Prljavobijela krutina, talište 192-194°C (raspad), Rf 0,33 (EtAc/heptan 1:2 s 1% TEA). 1H-NMR (DMSO-d6) 6.46 (br s, 3H), 6.53 (s, 1H), 6.65 (d od d, J1=2.3, J2=8.4, 1H), 6.89 (d, J=2.2, 1H), 7.35 (d, J=8.6, 1H), 7.88 (s, IH). MS (EI) m/z 337 (M+, 73). Analitički izračunato za C12H7Cl4NS·HCl: C, 38.38; H, 2.15; N, 3.73. Nađeno: C, 38.73; H, 2.07; N, 3.60. Off-white solid, mp 192-194°C (dec), Rf 0.33 (EtAc/heptane 1:2 with 1% TEA). 1H-NMR (DMSO-d6) 6.46 (br s, 3H), 6.53 (s, 1H), 6.65 (d of d, J1=2.3, J2=8.4, 1H), 6.89 (d, J=2.2, 1H) , 7.35 (d, J=8.6, 1H), 7.88 (s, 1H). MS (EI) m/z 337 (M+, 73). Analytical calculated for C12H7Cl4NS·HCl: C, 38.38; H, 2.15; N, 3.73. Found: C, 38.73; H, 2.07; N, 3.60.

3-Metoksi-4-(2,3-diklorfenilsulfanil}-fenilamin 3-Methoxy-4-(2,3-dichlorophenylsulfanyl}-phenylamine).

2,3-diklortiofenol + 2-brom-5-nitroanizol-4-nitro-2-metoksifenil-(2',3'-diklorfenil)-sulfid 2,3-dichlorothiophenol + 2-bromo-5-nitroanisole-4-nitro-2-methoxyphenyl-(2',3'-dichlorophenyl)-sulfide

Ovaj spoj je izdvojen kristaliziranjem iz etera, a ne dobivanjem HCl soli. Dobiveni su prljavobijeli kristali, talište 166-167°C, Rf 0,17 (EtAc/heptan 1:2 s 1% TEA). 1H-NMR (DMSO-d6) 3.67 (s, 3H), 5.75 (br s, 2H), 6.26 (d od d, J1=1.9, J2=8.3, 1 H), 6.37 (d, J=1.9, 1H), 6.46 (d, J=7.9, 1H), 7.13 (d, J=8.2, 1H), 7.16 (t, J=8.1, 1H), 7.31 (d, J=7.9, 1H). MS (EI) m/z 299 M+ [100]. Analitički izračunato za C13H11Cl2NOS: C, 52.01; H, 3.69; N, 4.67. Nađeno: C, 51.97; H, 3.59; N, 4.67. This compound was isolated by crystallization from ether, not by obtaining the HCl salt. Off-white crystals were obtained, melting point 166-167°C, Rf 0.17 (EtAc/heptane 1:2 with 1% TEA). 1H-NMR (DMSO-d6) 3.67 (s, 3H), 5.75 (br s, 2H), 6.26 (d of d, J1=1.9, J2=8.3, 1H), 6.37 (d, J=1.9, 1H ), 6.46 (d, J=7.9, 1H), 7.13 (d, J=8.2, 1H), 7.16 (t, J=8.1, 1H), 7.31 (d, J=7.9, 1H). MS (EI) m/z 299 M+ [100]. Analytical calculated for C13H11Cl2NOS: C, 52.01; H, 3.69; N, 4.67. Found: C, 51.97; H, 3.59; N, 4.67.

5-Amino-2-(2,3-diklorfenilsulfanil)-acetofenon hidroklorid 5-Amino-2-(2,3-dichlorophenylsulfanyl)-acetophenone hydrochloride

2,3-diklortiofenol + 2-klornitroacetofenon- 4-nitro-2-acetilfenil-(2',3'-diklorfenil)-sulfid 2,3-dichlorothiophenol + 2-chloronitroacetophenone-4-nitro-2-acetylphenyl-(2',3'-dichlorophenyl)-sulfide

Žuta krutina, talište 151-153ºC (raspad), Rf 0,35 (EtAc/heptan 1:1 s 1% TEA). 1H-NMR (DMSO-d6) 2.51 (s, 3H), 7.00-7.09 (m, 3H), 7.34 (t, J=7.9, 1H), 7.41 (brs, 1H), 7.44 (br s, 3H), 7.58 (d, J=7.9, 1H). MS (EI) m/z 311 M+ [100]. Analitički izračunato za C13H11Cl3NOS·HCl: C, 48.23; H, 3.47; N, 4.02. Nađeno: C, 47.78; H, 3.43; N, 3.79. Yellow solid, mp 151-153ºC (dec), Rf 0.35 (EtAc/heptane 1:1 with 1% TEA). 1H-NMR (DMSO-d6) 2.51 (s, 3H), 7.00-7.09 (m, 3H), 7.34 (t, J=7.9, 1H), 7.41 (brs, 1H), 7.44 (br s, 3H), 7.58 (d, J=7.9, 1H). MS (EI) m/z 311 M+ [100]. Analytical calculated for C13H11Cl3NOS·HCl: C, 48.23; H, 3.47; N, 4.02. Found: C, 47.78; H, 3.43; N, 3.79.

4-(2,3-diklorfenilsulfanil)-fenilamin 4-(2,3-dichlorophenylsulfanyl)-phenylamine

2,3-diklortiofenol + 4-bromnitrofenol-4-nitrofenil-(2',3'-diklorfenil)sulfid 2,3-dichlorothiophenol + 4-bromonitrophenol-4-nitrophenyl-(2',3'-dichlorophenyl)sulfide

Prljavobijela krutina, talište 175°C (raspad), Rf 0,31 (EtAc/heptan 1:2 s 1 % TEA). 1H-NMR (DMSO-d6) 7.13 (d, J=8.2, 2H); 7.26 (t, J=7.8, 1H), 7.42 (d, J=8.6, 2H), 7.47 (d od d, J1=1.9, J2=7.8, 1H), 7.76 (d od d, J1=1.3, J2=7.9, 1H), 8.04 (br s, 3H). MS (EI) m/z 269 M+ [100]. Analitički izračunato za C12H9Cl2NS·HCl: C, 47.00; H. 3.29; N, 4.57. Nađeno: C, 46.92; H, 3.36; N, 4.27. Off-white solid, mp 175°C (dec), Rf 0.31 (EtAc/heptane 1:2 with 1% TEA). 1H-NMR (DMSO-d 6 ) 7.13 (d, J=8.2, 2H); 7.26 (t, J=7.8, 1H), 7.42 (d, J=8.6, 2H), 7.47 (d from d, J1=1.9, J2=7.8, 1H), 7.76 (d from d, J1=1.3, J2 =7.9, 1H), 8.04 (br s, 3H). MS (EI) m/z 269 M+ [100]. Analytical calculated for C12H9Cl2NS·HCl: C, 47.00; H. 3.29; N, 4.57. Found: C, 46.92; H, 3.36; N, 4.27.

3-Klor-4-(1-naftilsulfanil~fenilamin hidroklorid 3-Chloro-4-(1-naphthylsulfanyl~phenylamine hydrochloride).

1-naftalentiol + 3,4-diklornitrobenzen-4-nitro-2-klorfenil-(1-naftil)-sulfid 1-naphthalenethiol + 3,4-dichloronitrobenzene-4-nitro-2-chlorophenyl-(1-naphthyl)-sulfide

Prljavobijela krutina, talište 192-194°C, Rf 0,37 (EtAc/heptan 1:2 s 1% TEA). 1H-NMR (DMSO-d6) 6.84 (m, 2H), 7.20 (d, J=1.9, 1H), 7.40 (d, J=7.0, 1H), 7.50 (t, J=7.8, 1H), 7.56-7.61 (m, 2H), 7.94 (d, J=8.2, 1H), 7.99 (m, 1H), 8.12-8.15 (m, 5H). MS (EI) m/z 285 M+ [100]. Analitički izračunato za C16H12ClNS·HCl: C, 59.64; H, 4.07; N, 4.35. Nađeno: C, 59.59; H, 4.19; N, 4.11. Off-white solid, mp 192-194°C, Rf 0.37 (EtAc/heptane 1:2 with 1% TEA). 1H-NMR (DMSO-d6) 6.84 (m, 2H), 7.20 (d, J=1.9, 1H), 7.40 (d, J=7.0, 1H), 7.50 (t, J=7.8, 1H), 7.56- 7.61 (m, 2H), 7.94 (d, J=8.2, 1H), 7.99 (m, 1H), 8.12-8.15 (m, 5H). MS (EI) m/z 285 M+ [100]. Analytical calculated for C16H12ClNS·HCl: C, 59.64; H, 4.07; N, 4.35. Found: C, 59.59; H, 4.19; N, 4.11.

3-Metil-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 3-Methyl-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride

2,4-diklortiofenol + 2-brom-5-nitrotoluen-4-nitro-2-metilfenil-(2', 4'-diklorfenil)-sulfid 2,4-dichlorothiophenol + 2-bromo-5-nitrotoluene-4-nitro-2-methylphenyl-(2', 4'-dichlorophenyl)-sulfide

Prljavobijela krutina, talište 195-197°C, Rf 0,41 (EtAc/heptan 1:2 s 1% TEA). 1H-NMR (DMSO-d6) 2.22 (s. 3H), 6.62 (d, J=8.6, 1H), 6.94 (d, J=8.2, 1H), 7.04 (br s, 1H), 7.29 (s, 1H), 7.32 (s, 1H), 7.66 (d, J=2.2, 1H), 7.89 (br s, 3H). MS (EI) m/z 283 M+ [100]. Analitički izračunato za C13H11Cl2NS·HCl: C, 48.69; H, 3.77; N, 4.37. Nađeno: C, 48.87; H, 3.73; N, 4.34. Off-white solid, mp 195-197°C, Rf 0.41 (EtAc/heptane 1:2 with 1% TEA). 1H-NMR (DMSO-d6) 2.22 (s. 3H), 6.62 (d, J=8.6, 1H), 6.94 (d, J=8.2, 1H), 7.04 (br s, 1H), 7.29 (s, 1H ), 7.32 (s, 1H), 7.66 (d, J=2.2, 1H), 7.89 (br s, 3H). MS (EI) m/z 283 M+ [100]. Analytical calculated for C13H11Cl2NS·HCl: C, 48.69; H, 3.77; N, 4.37. Found: C, 48.87; H, 3.73; N, 4.34.

3-Brom-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 3-Bromo-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride

2,4-diklortiofenol + 3-brom-4-klornitrobenzen-4-nitro-2-bromfenil-(2', 4'-diklorfenil)-sulfid 2,4-dichlorothiophenol + 3-bromo-4-chloronitrobenzene-4-nitro-2-bromophenyl-(2', 4'-dichlorophenyl)-sulfide

Prljavobijela krutina, talište 171-173°C (raspad), Rf 0,69 (EtAc/heptan 1:1 s 1% TEA). 1H-NMR (DMSO-d6) 6.66 (d, J=8.6, 1H), 6.81 (d od d, J1=2.2, J2=8.4, 1H), 7.19 (d, J=2.2, 1H), 7.29 (d, J=8.2, 1H), 7.32 (d od d, J1=2.2, J2=8.6, 1H), 7.64 (d, J=2.2, 1H), 7.92 (br s, 3H). MS (EI) m/z 347 (M+, 60). Analitički izračunato za C12H7BrCl2NS·HCl: C, 37.39; H, 2.35; N, 3.63. Nađeno: C, 37.78; H, 2.28; N, 3.61. Off-white solid, mp 171-173°C (dec), Rf 0.69 (EtAc/heptane 1:1 with 1% TEA). 1H-NMR (DMSO-d6) 6.66 (d, J=8.6, 1H), 6.81 (d from d, J1=2.2, J2=8.4, 1H), 7.19 (d, J=2.2, 1H), 7.29 (d , J=8.2, 1H), 7.32 (d of d, J1=2.2, J2=8.6, 1H), 7.64 (d, J=2.2, 1H), 7.92 (br s, 3H). MS (EI) m/z 347 (M+, 60). Analytical calculated for C12H7BrCl2NS·HCl: C, 37.39; H, 2.35; N, 3.63. Found: C, 37.78; H, 2.28; N, 3.61.

2,5-Diklor-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 2,5-Dichloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride

2,4-diklortiofenol + 2,4,5-triklornitrofenol - 4-nitro-2,5-diklorfenil-(2', 4'-diklorfenil)-sulfid 2,4-dichlorothiophenol + 2,4,5-trichloronitrophenol - 4-nitro-2,5-dichlorophenyl-(2', 4'-dichlorophenyl)-sulfide

Prljavobijela krutina, talište 168-176°C, Rf 0,39 (EtAc/heptan 1:2 s 1% TEA). 1H-NMR (DMSO-d6) 6.57 (d, J=8.6, 1H), 6.83 (br s, 3H), 7.06 (s, 1H), 7.30 (d od d, J1= 2.2, J2=8.6, 1H), 7.55 (s, 1H), 7.63 (d, J=2.2, 1H). MS (EI) m/z 347 (M+, 60). Analitički izračunato za C12H7Cl4NS·0,75HCl: C, 39.34; H, 2.13; N, 3.82. Nađeno: C, 39.34; H, 2.11; N, 3.71. Off-white solid, mp 168-176°C, Rf 0.39 (EtAc/heptane 1:2 with 1% TEA). 1H-NMR (DMSO-d6) 6.57 (d, J=8.6, 1H), 6.83 (br s, 3H), 7.06 (s, 1H), 7.30 (d of d, J1= 2.2, J2=8.6, 1H) , 7.55 (s, 1H), 7.63 (d, J=2.2, 1H). MS (EI) m/z 347 (M+, 60). Analytical calculated for C12H7Cl4NS·0.75HCl: C, 39.34; H, 2.13; N, 3.82. Found: C, 39.34; H, 2.11; N, 3.71.

4,5-Diklor-2-(2,4-diklorfenilsulfanil)-fenilamin 4,5-Dichloro-2-(2,4-dichlorophenylsulfanyl)-phenylamine

2,4-diklortiofenol + 3,4-diklor-2-fluornitrobenzen-4-nitro-2-klor-5-fluorfenil-(2', 4'-diklorfenil)-sulfid 2,4-dichlorothiophenol + 3,4-dichloro-2-fluoronitrobenzene-4-nitro-2-chloro-5-fluorophenyl-(2', 4'-dichlorophenyl)-sulfide

Polazeći od 2-fluornitrobenzena, zamijenjen je fluor da se dobije 2-sulfanilni spoj. Ovaj spoj je pročišćen “flash” kromatografijom kao slobodni amin, prljavobijela krutina, talište 119-122°C, Rf 0,20 (EtAc/heptan 1:20). 1H-NMR (DMSO-d6) 5.93 (s, 2H), 6.61 (d, J=8.6, 1H), 7.08 (s, 1H), 7.31 (d od d, J1=2.1, J2=8.5, 1H), 7.55 (s, 1H), 7.65 (d, J=1.9, 1H). MS (EI) m/z 337 (M+, 75). Analitički izračunato za C12H7Cl4NS: C, 42.51; H, 2.08; N, 4.13. Nađeno: C, 42.94; H, 1.97; N, 4.08. Starting from 2-fluoronitrobenzene, the fluorine was replaced to give the 2-sulfanyl compound. This compound was purified by flash chromatography as the free amine, off-white solid, mp 119-122°C, Rf 0.20 (EtAc/heptane 1:20). 1H-NMR (DMSO-d6) 5.93 (s, 2H), 6.61 (d, J=8.6, 1H), 7.08 (s, 1H), 7.31 (d from d, J1=2.1, J2=8.5, 1H), 7.55 (s, 1H), 7.65 (d, J=1.9, 1H). MS (EI) m/z 337 (M+, 75). Analytical calculated for C12H7Cl4NS: C, 42.51; H, 2.08; N, 4.13. Found: C, 42.94; H, 1.97; N, 4.08.

3,5-Diklor-4-(2,4-diklorfenilsulfanil)-fenilamin 3,5-Dichloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine

2,4-diklortiofenol + 3,4,5-triklornitrobenzen-4-nitro-2, 6-diklorfenil-(2', 4'-diklorfenil)-sulfid 2,4-dichlorothiophenol + 3,4,5-trichloronitrobenzene-4-nitro-2, 6-dichlorophenyl-(2', 4'-dichlorophenyl)-sulfide

Ovaj spoj je rekristaliziran kao slobodni amin da se dobije bijela krutina, talište 128-131°C, Rf 0,36 (EtAc/heptan 1:2). 1H-NMR (DMSO-d6) 6.33 (br s, 2H), 6.46 (d, J=8.5, 1H), 6.83 (s, 2H), 7.32 (d od d, J1=2.1, J2=8.5, 1H), 7.65 (d, J=2.2, 1H). MS (EI) m/z 337 (M+, 77). Analitički izračunato za C12H7Cl4NS: C, 42.51; H, 2.08; N, 4.13. Nađeno: C, 42.15; H, 2.10; N, 4.01. This compound was recrystallized as the free amine to give a white solid, mp 128-131°C, Rf 0.36 (EtAc/heptane 1:2). 1H-NMR (DMSO-d6) 6.33 (br s, 2H), 6.46 (d, J=8.5, 1H), 6.83 (s, 2H), 7.32 (d of d, J1=2.1, J2=8.5, 1H) , 7.65 (d, J=2.2, 1H). MS (EI) m/z 337 (M+, 77). Analytical calculated for C12H7Cl4NS: C, 42.51; H, 2.08; N, 4.13. Found: C, 42.15; H, 2.10; N, 4.01.

2,3-Diklor-4-(2,4-diklorfenilsulfanil)-fenilamin 2,3-Dichloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine

2,4-diklortiofenol + 2,3,4-trinitrobenzen-2,3-diklor-4-(2,4-diklorfeniltio)-nitrobenzen 2,4-dichlorothiophenol + 2,3,4-trinitrobenzene-2,3-dichloro-4-(2,4-dichlorophenylthio)-nitrobenzene

Ovaj spoj je pročišćen “flash” kromatografijom kao slobodni amin da se dobije bijela krutina, talište 116-119°C, Rf 0,33 (EtAc/heptan 1:4 s 1% TEA). 1H-NMR (DMSO-d6) 6.09 (s, 2H), 6.47 (d, J=8.7, 1H), 6.86 (d, J=8.7, 1H), 7.32 (d od d, J1=2.3, J2=8.7, 1H), 7.46 (d, J=8.7, 1H), 7.69 (d, J=2.2, 1H). MS (EI) m/z 337 (M+, 71). Analitički izračunato za C12H7Cl4NS: C, 42.51; H, 2.08; N, 4.13. Nađeno: C, 42.83; H, 2.02; N, 4.06. This compound was purified by flash chromatography as the free amine to give a white solid, mp 116-119°C, Rf 0.33 (EtAc/heptane 1:4 with 1% TEA). 1H-NMR (DMSO-d6) 6.09 (s, 2H), 6.47 (d, J=8.7, 1H), 6.86 (d, J=8.7, 1H), 7.32 (d from d, J1=2.3, J2=8.7 , 1H), 7.46 (d, J=8.7, 1H), 7.69 (d, J=2.2, 1H). MS (EI) m/z 337 (M+, 71). Analytical calculated for C12H7Cl4NS: C, 42.51; H, 2.08; N, 4.13. Found: C, 42.83; H, 2.02; N, 4.06.

B. Opća sintetska metoda B B. General synthetic method B

Opća sintetska metoda B je dijagramom prikazana niže: The general synthetic method B is shown diagrammatically below:

[image] [image]

gdje je X halogen, poželjno klor: where X is halogen, preferably chlorine:

Sljedeći spojevi su priređeni korištenjem opće sintetske metode B: The following compounds were prepared using general synthetic method B:

4-Nitro-2-klorfenil-(2',4'-dimetilfenil)-sulfid 4-Nitro-2-chlorophenyl-(2',4'-dimethylphenyl)-sulfide

4-Amino-2-klorfenil-(2',4'-dimetilfenil)-sulfid hidroklorid 4-Amino-2-chlorophenyl-(2',4'-dimethylphenyl)-sulfide hydrochloride

4-Amino-2-klorfenil-(2'-metil-4'-klorfenil)-sulfid 4-Amino-2-chlorophenyl-(2'-methyl-4'-chlorophenyl)-sulfide

4-Amino-2-klorfenil-(2',4'-difluorfenil)-sulfid hidroklorid 4-Amino-2-chlorophenyl-(2',4'-difluorophenyl)-sulfide hydrochloride

4-amino-2-klorfenil-(2',4',6'-triklorfenil)-sulfid 4-amino-2-chlorophenyl-(2',4',6'-trichlorophenyl)sulfide

4-Amino-2-klorfenil-(2'-amino-4'-klorfenil)-sulfid 4-Amino-2-chlorophenyl-(2'-amino-4'-chlorophenyl)-sulfide

4-Amino-2-klorfenil-(3',4'-diklorfenil)-sulfid 4-Amino-2-chlorophenyl-(3',4'-dichlorophenyl)-sulfide

4-Amino-2-klorfenil-2-(3-klor-5-trifluormetilpiridil)-sulfid 4-Amino-2-chlorophenyl-2-(3-chloro-5-trifluoromethylpyridyl)-sulfide

Bis-(4,4'-diamino-2,2'-diklorfenil)-sulfid Bis-(4,4'-diamino-2,2'-dichlorophenyl)-sulfide

4-Amino-2-klorfenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-chlorophenyl-(2',4'-dichlorophenyl)-sulfide

2-Klor-4-amino-5-metilaminofenil-(2',4'-diklorfenil)-sulfid 2-Chloro-4-amino-5-methylaminophenyl-(2',4'-dichlorophenyl)-sulfide

2-Klor-4-amino-5-morfolinofenil-(2',4'-diklorfenil)-sulfid 2-Chloro-4-amino-5-morpholinophenyl-(2',4'-dichlorophenyl)-sulfide

4-Amino-2-trifluormetilfenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-trifluoromethylphenyl-(2',4'-dichlorophenyl)-sulfide

4-Amino-2-fluorfenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-fluorophenyl-(2',4'-dichlorophenyl)-sulfide

5-Amino-3-klorfenil-(2',4'-diklorfenil)-sulfid 5-Amino-3-chlorophenyl-(2',4'-dichlorophenyl)-sulfide

Početni stupanj opće sintetske metode B prikazan je priređivanjem intermedijernog spoja koje je niže opisano. The initial step of the general synthetic method B is shown by the preparation of the intermediate compound described below.

4-Nitro-2-klorfenol-(2',4'dimetilfenil)-sulfid 4-Nitro-2-chlorophenol-(2',4'dimethylphenyl)sulfide

2,4-Dimetiltiofenol (1,0 g) i 3,4-diklornitrobenzen (1 ekv.) dodani su u 100 ml acetona koji sadrži K2CO3 (5 g). Smjesa je refluksirana 24 h. Nakon hlađenja na sobnu temperaturu, smjesa je filtrirana i aceton je uklonjen rotacijskim vakuumom. Dobivene rezidue su otopljene u malom volumenu CH2Cl2 i filtrirane. CH2Cl2 je zatim uklonjen pomoću rotacijskog vakuuma. Dobivene rezidue su tretirane metanolom (MeOH), što je izazvalo taloženje produkta. Žuti čvrsti produkt zatim je sakupljen filtriranjem i osušen na 40°C u vakuumu tijekom 24 h. Prinos produkta bio je 66%, a talište je bilo 128-130°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 2,4-Dimethylthiophenol (1.0 g) and 3,4-dichloronitrobenzene (1 eq.) were added to 100 ml of acetone containing K 2 CO 3 (5 g). The mixture was refluxed for 24 h. After cooling to room temperature, the mixture was filtered and the acetone was removed by rotary vacuum. The obtained residues were dissolved in a small volume of CH2Cl2 and filtered. The CH2Cl2 was then removed using a rotary vacuum. The obtained residues were treated with methanol (MeOH), which caused precipitation of the product. The yellow solid was then collected by filtration and dried at 40°C under vacuum for 24 h. The product yield was 66%, and the melting point was 128-130°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

Sljedeći spojevi su priređeni koristeći početni stupanj sukladno gore prikazanom intermedijeru. The following compounds were prepared using the starting step according to the intermediate shown above.

4-Amino-2-klorfenil-(2',4'-dimetilfenil)-sulfid hidroklorid 4-Amino-2-chlorophenyl-(2',4'-dimethylphenyl)-sulfide hydrochloride

2-Klor-4-nitrofenil-(2',4'-dimetilfenil)-sulfid (0,85 g) i željezni prah u MeOH (300 ml) dodani su u 20 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform:heksan (1:4). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Prljavobijela krutina je sakupljena filtriranjem i osušena na 50°C u vakuumu tijekom 24 h. Prinos produkta bio je 57%, a talište je bilo 180-185°C (raspad). Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 2-Chloro-4-nitrophenyl-(2',4'-dimethylphenyl)sulfide (0.85 g) and iron powder in MeOH (300 ml) were added to 20 ml of aqueous NH4Cl, in a molar ratio of 1:3: 5 for the substrate, iron powder, or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh) and then elution using the solvent system chloroform:hexane (1:4). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The off-white solid was collected by filtration and dried at 50°C under vacuum for 24 h. The product yield was 57%, and the melting point was 180-185°C (decomposition). Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-Amino-2-klorfenil-(2'-metil-4'-klorfenil-sulfid 4-Amino-2-chlorophenyl-(2'-methyl-4'-chlorophenyl-sulphide).

4-Nitro-2-klorfenil-(2'-metil-4'-klorfenil)-sulfid (1,0 g) i željezni prah u MeOH (300 ml) dodani su u 5 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malenom volumenu smjese kloroform:heksan (1:4) te filtrirane. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform:heksan (1:1). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Bijeli čvrsti produkt je sakupljen filtriranjem i osušen na 60°C u vakuumu tijekom 24 h. Prinos produkta bio je 71%, a talište je bilo 91-93°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 4-Nitro-2-chlorophenyl-(2'-methyl-4'-chlorophenyl)sulfide (1.0 g) and iron powder in MeOH (300 ml) were added to 5 ml of aqueous NH4Cl, in a molar ratio of 1: 3:5 for the substrate, iron powder, or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of the mixture of chloroform:hexane (1:4) and filtered. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh) and then elution using the solvent system chloroform:hexane (1:1). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The white solid product was collected by filtration and dried at 60°C under vacuum for 24 h. The product yield was 71%, and the melting point was 91-93°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-Amino-2-klorfenil-(2',4'-difluorfenil)-sulfid hidroklorid 4-Amino-2-chlorophenyl-(2',4'-difluorophenyl)-sulfide hydrochloride

4-Nitro-2-klorfenil-(2',4'-difluorfenil)-sulfid i željezni prah u MeOH (300 ml) dodani su u 20 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform:heksan (1:5). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Bezbojni čvrsti produkt sakupljen je filtriranjem i osušen na 50°C u vakuumu tijekom 24 h. Prinos produkta bio je 88%, a talište je bilo 187-190°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 4-Nitro-2-chlorophenyl-(2',4'-difluorophenyl)-sulfide and iron powder in MeOH (300 ml) were added to 20 ml of aqueous NH4Cl, in a molar ratio of 1:3:5 for substrate, iron powder , or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh) and then elution using the solvent system chloroform:hexane (1:5). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The colorless solid product was collected by filtration and dried at 50°C under vacuum for 24 h. The product yield was 88%, and the melting point was 187-190°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-amino-2-klorfenil-(2',4',6'-triklorfenil)-sulfid 4-amino-2-chlorophenyl-(2',4',6'-trichlorophenyl)sulfide

4-Nitro-2-klorfenil-(2',4',6;-triklorfenil)-sulfid (0,6 g) i željezni prah u MeOH (250 ml) dodani su u 32 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform:heksan (1:3). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Bijeli čvrsti produkt je sakupljen filtriranjem i osušen na 40°C u vakuumu tijekom 24 h. Prinos produkta bio je 72%, a talište je bilo 109-111°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 4-Nitro-2-chlorophenyl-(2',4',6;-trichlorophenyl)-sulfide (0.6 g) and iron powder in MeOH (250 ml) were added to 32 ml of aqueous NH4Cl, in a molar ratio of 1 :3:5 for the substrate, iron powder, or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh) and then eluting using the solvent system chloroform:hexane (1:3). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The white solid product was collected by filtration and dried at 40°C under vacuum for 24 h. The product yield was 72%, and the melting point was 109-111°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-Amino-2-klorfenil-(2'-amino-4'-klorfenil)-sulfid 4-Amino-2-chlorophenyl-(2'-amino-4'-chlorophenyl)-sulfide

4-Amino-2-klorfenil-(2'-nitro-4'-klorfenil)-sulfid (1,02 g) i željezni prah u MeOH (250 ml) dodani su u 60 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform:heksan (3:7). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Prljavobijela krutina je sakupljena filtriranjem i osušena na 50°C u vakuumu tijekom 24 h. Prinos produkta bio je 59%, a talište je bilo 86-88ºC. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 4-Amino-2-chlorophenyl-(2'-nitro-4'-chlorophenyl)sulfide (1.02 g) and iron powder in MeOH (250 ml) were added to 60 ml of aqueous NH4Cl, in a molar ratio of 1: 3:5 for the substrate, iron powder, or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh), and then eluting using the solvent system chloroform:hexane (3:7). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The off-white solid was collected by filtration and dried at 50°C under vacuum for 24 h. The product yield was 59%, and the melting point was 86-88ºC. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-Amino-2-klorfenil-(3',4'-diklorfenil)-sulfid 4-Amino-2-chlorophenyl-(3',4'-dichlorophenyl)-sulfide

4-Nitro-2-klorfenil-(3',4'-diklorfenil)-sulfid (1,0 g) i željezni prah u MeOH (250 ml) dodani su u 40 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform:heksan (3:7). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Bezbojni čvrsti produkt je sakupljen filtriranjem i osušen na 70°C u vakuumu tijekom 24 h. Talište produkta je bilo 103-105°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 4-Nitro-2-chlorophenyl-(3',4'-dichlorophenyl)-sulfide (1.0 g) and iron powder in MeOH (250 ml) were added to 40 ml of aqueous NH4Cl, in a molar ratio of 1:3: 5 for the substrate, iron powder, or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh), and then eluting using the solvent system chloroform:hexane (3:7). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The colorless solid product was collected by filtration and dried at 70°C under vacuum for 24 h. The melting point of the product was 103-105°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-Amino-2-klorfenil-2-(3-klor-5-trifluormetilpiridil-sulfid 4-Amino-2-chlorophenyl-2-(3-chloro-5-trifluoromethylpyridyl-sulfide).

4-Nitro-2-klorfenil-2-(3-klor-5-trifluormetilpiridil)-sulfid (1,5 g) i željezni prah u MeOH (250 ml) dodani su u 20 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform:heksan (3:7). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Bezbojni čvrsti produkt je sakupljen filtriranjem i osušen na 60ºC u vakuumu tijekom 24 h. Prinos produkta bio je 97%, a talište je bilo 96-98°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 4-Nitro-2-chlorophenyl-2-(3-chloro-5-trifluoromethylpyridyl)sulfide (1.5 g) and iron powder in MeOH (250 ml) were added to 20 ml of aqueous NH4Cl, in a molar ratio of 1: 3:5 for the substrate, iron powder, or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh), and then eluting using the solvent system chloroform:hexane (3:7). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The colorless solid product was collected by filtration and dried at 60ºC under vacuum for 24 h. The product yield was 97%, and the melting point was 96-98°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

Bis-(4,4'-diamino-2,2'-diklorfenil)-sulfid Bis-(4,4'-diamino-2,2'-dichlorophenyl)-sulfide

4-Amino-2-klorfenil-(2'-klor-4'-nitrofenil)- i željezni prah u MeOH (250 ml) dodani su u 20 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform:heksan (3:7). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Prljavobijeli produkt je sakupljen filtriranjem i osušen na 60°C u vakuumu tijekom 24 h. Prinos produkta bio je 66%, a talište je bilo 113-115 °C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 4-Amino-2-chlorophenyl-(2'-chloro-4'-nitrophenyl)- and iron powder in MeOH (250 ml) were added to 20 ml of aqueous NH4Cl, in a molar ratio of 1:3:5 for substrate, iron powder, or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh), and then eluting using the solvent system chloroform:hexane (3:7). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The off-white product was collected by filtration and dried at 60°C in vacuum for 24 h. The product yield was 66%, and the melting point was 113-115 °C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-Amino-2-klorfenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-chlorophenyl-(2',4'-dichlorophenyl)-sulfide

2-Klor-4-nitrofenil-(2',4'-diklorfenil)-sulfid i željezni prah u MeOH (250 ml) dodani su u 20 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane, pa je otapalo uklonjeno iz filtrata rotacijskim uparivačem. Produkt je dobiven miješanjem rezidua sa 75 ml destilirane vode (dH2O). Bezbojni čvrsti talog je sakupljen filtriranjem. Prinos produkta bio je 83%, a talište je bilo 105-107°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 2-Chloro-4-nitrophenyl-(2',4'-dichlorophenyl)-sulfide and iron powder in MeOH (250 ml) were added to 20 ml of aqueous NH4Cl, in a molar ratio of 1:3:5 for substrate, iron powder , or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered, and the solvent was removed from the filtrate using a rotary evaporator. The product was obtained by mixing the residue with 75 ml of distilled water (dH2O). A colorless solid precipitate was collected by filtration. The product yield was 83%, and the melting point was 105-107°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-Amino-2-klorfenil-(4'-acetamido-2'-klorfenil)-sulfid 4-Amino-2-chlorophenyl-(4'-acetamido-2'-chlorophenyl)-sulfide

4-Nitro-2-klorfenil-(4'-acetamido-2'-klorfenil)-sulfid i željezni prah u MeOH (250 ml) dodani su u 20 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform-heksan (1:4). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Prljavobijela krutina sakupljena je filtriranjem i osušena na 60°C u vakuumu tijekom 24 h. Prinos produkta bio je 76%, a talište je bilo 170-175 °C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 4-Nitro-2-chlorophenyl-(4'-acetamido-2'-chlorophenyl)-sulfide and iron powder in MeOH (250 ml) were added to 20 ml of aqueous NH4Cl, in a molar ratio of 1:3:5 for the substrate, iron powder, or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh) and then elution using the solvent system chloroform-hexane (1:4). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The off-white solid was collected by filtration and dried at 60°C under vacuum for 24 h. The product yield was 76%, and the melting point was 170-175 °C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-Amino-2-klorfenil-(4'-dimetilamino-2'-florfenil-sulfid 4-Amino-2-chlorophenyl-(4'-dimethylamino-2'-fluorophenyl-sulfide).

4-Nitro-2-klorfenil-(4'-dimetilamino-2'-klorfenil)-sulfid (0.2 g) i željezni prah u MeOH (250 ml) dodani su u 20 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform:heksan (2:3). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Blijedožuti čvrst produkt je sakupljen filtriranjem i osušen na 60°C u vakuumu tijekom 24 h. Prinos produkta bio je 58%. a talište je bilo 133-135°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 4-Nitro-2-chlorophenyl-(4'-dimethylamino-2'-chlorophenyl)-sulfide (0.2 g) and iron powder in MeOH (250 ml) were added to 20 ml of aqueous NH4Cl solution, in a molar ratio of 1:3: 5 for the substrate, iron powder, or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh) and then elution using the solvent system chloroform:hexane (2:3). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The pale yellow solid product was collected by filtration and dried at 60°C under vacuum for 24 h. The product yield was 58%. and the melting point was 133-135°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

2-Klor-4-amino-5-metilaminofenil-(2',4'-diklorfenil)-sulfid 2-Chloro-4-amino-5-methylaminophenyl-(2',4'-dichlorophenyl)-sulfide

2-Klor-4-nitro-5-metilaminofenil-(2',4'-diklorfenil)-sulfid (0.2 g) i željezni prah u MeOH (250 ml) dodani su u 20 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform:heksan (1:4). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Smeđi čvrsti produkt je sakupljen filtriranjem i osušen na 50°C u vakuumu tijekom 24 h. Prinos produkta bio je 16%, a talište je bilo 65-70°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 2-Chloro-4-nitro-5-methylaminophenyl-(2',4'-dichlorophenyl)-sulfide (0.2 g) and iron powder in MeOH (250 ml) were added to 20 ml of aqueous NH4Cl, in a molar ratio of 1: 3:5 for the substrate, iron powder, or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh) and then elution using the solvent system chloroform:hexane (1:4). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The brown solid product was collected by filtration and dried at 50°C under vacuum for 24 h. The product yield was 16%, and the melting point was 65-70°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

2-Klor-4-amino-5-N-morfolinofenil-(2',4'-diklorfenil)-sulfid 2-Chloro-4-amino-5-N-morpholinophenyl-(2',4'-dichlorophenyl)-sulfide

2-Klor-4-nitro-5-N-morfolinofenil-(2',4'-diklorfenil)-sulfid (0,9 g) i željezni prah u MeOH (250 ml) dodani su u 20 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform:heksan (3:2). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Bijeli čvrsti produkt je sakupljen filtriranjem i osušen na 50°C u vakuumu tijekom 24 h. Talište produkta je bilo 153-155°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 2-Chloro-4-nitro-5-N-morpholinophenyl-(2',4'-dichlorophenyl)sulfide (0.9 g) and iron powder in MeOH (250 ml) were added to 20 ml of aqueous NH4Cl, in with a molar ratio of 1:3:5 for the substrate, iron powder, or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh) and then elution using the solvent system chloroform:hexane (3:2). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The white solid product was collected by filtration and dried at 50°C under vacuum for 24 h. The melting point of the product was 153-155°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-Amino-2-trifluormetilfenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-trifluoromethylphenyl-(2',4'-dichlorophenyl)-sulfide

4-Nitro-2-trifluormetilfenil-(2',4'-diklorfenil)-sulfid (0,6 g) i željezni prah u MeOH (250 ml) dodani su u 20 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform:heksan (1:1). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Produkt je sakupljen filtriranjem i osušen na 50°C u vakuumu tijekom 24 h. Talište produkta nije određeno jer je produkt bezbojno ulje. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 4-Nitro-2-trifluoromethylphenyl-(2',4'-dichlorophenyl)-sulfide (0.6 g) and iron powder in MeOH (250 ml) were added to 20 ml of aqueous NH4Cl, in a molar ratio of 1:3: 5 for the substrate, iron powder, or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh) and then elution using the solvent system chloroform:hexane (1:1). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The product was collected by filtration and dried at 50°C in a vacuum for 24 h. The melting point of the product is not determined because the product is a colorless oil. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-Amino-2-fluorfenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-fluorophenyl-(2',4'-dichlorophenyl)-sulfide

4-Nitro-2-fluorfenil-(2',4'-diklorfenil)-sulfid i željezni prah u MeOH (250 ml) dodani su u 20 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform:heksan (1:1). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Blijedožut čvrsti produkt sakupljen je filtriranjem i osušen na 30°C u vakuumu tijekom 24 h. Prinos produkta bio je 55%, a talište je bilo 101-102°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 4-Nitro-2-fluorophenyl-(2',4'-dichlorophenyl)-sulfide and iron powder in MeOH (250 ml) were added to 20 ml of aqueous NH4Cl, in a molar ratio of 1:3:5 for substrate, iron powder , or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh) and then elution using the solvent system chloroform:hexane (1:1). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The pale yellow solid product was collected by filtration and dried at 30°C under vacuum for 24 h. The product yield was 55%, and the melting point was 101-102°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

5-Amino-3-klorfenil-(2',4'-diklorfenil)-sulfid 5-Amino-3-chlorophenyl-(2',4'-dichlorophenyl)-sulfide

3-Klor-5-nitrofenil-(2',4'-diklorfenil)-sulfid (1,0 g) i željezni prah u MeOH (250 ml) dodani su u 20 ml vodene otopine NH4Cl, u molarnom odnosu 1:3:5 za supstrat, željezni prah, odnosno NH4Cl. Smjesa je preko noći miješana mehanički pod refluksom. Otapalo je uklonjeno korištenjem rotacijskog vakuuma. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform:heksan (3:2). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Produkt je sakupljen filtriranjem i osušen na 25°C u vakuumu tijekom 24 h. Prinos produkta bio je 16%, a talište nije određeno jer je produkt bio smeđe ulje. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 3-Chloro-5-nitrophenyl-(2',4'-dichlorophenyl)sulfide (1.0 g) and iron powder in MeOH (250 ml) were added to 20 ml of aqueous NH4Cl, in a molar ratio of 1:3: 5 for the substrate, iron powder, or NH4Cl. The mixture was stirred mechanically under reflux overnight. The solvent was removed using a rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh) and then elution using the solvent system chloroform:hexane (3:2). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The product was collected by filtration and dried at 25°C in a vacuum for 24 h. The yield of the product was 16%, and the melting point was not determined because the product was a brown oil. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

C. Opća sintetska metoda C C. General synthetic method C

Niže je shematski, dijagramom, prikazana opća sintetska metoda C: Below is a schematic diagram of the general synthetic method C:

[image] [image]

Općom sintetskom metodom C priređeni su sljedeći spojevi: The following compounds were prepared by general synthetic method C:

4-Amino-2-klorfenil-(2'-klor-4'-nitrofenil)-sulfid 4-Amino-2-chlorophenyl-(2'-chloro-4'-nitrophenyl)-sulfide

4-Amino-2-klorfenil-(2'-nitro-4'-klorfenil)-sulfid 4-Amino-2-chlorophenyl-(2'-nitro-4'-chlorophenyl)-sulfide

4-Amino-2-klorfenil-6-(5-nitrokinolino)-sulfid 4-Amino-2-chlorophenyl-6-(5-nitroquinolino)-sulfide

4-Amino-2-klorfenil-(2'-klor-4'-nitrofenil)-sulfid 4-Amino-2-chlorophenyl-(2'-chloro-4'-nitrophenyl)-sulfide

2-Klor-4-aminotiofenol (2,0 g) i 3,4-diklornitrobenzen (1 ekv.) dodani su u 300 m1 acetona koji sadrži K2CO3 (20 g). Smjesa je refluksirana 24 h na 60°C. Nakon hlađenja na sobnu temperaturu, smjesa je filtrirana i aceton je uklonjen rotacijskim vakuumom. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Kloroform je zatim uklonjen rotacijskim vakuumom. Dobivene rezidue su razmuljene s MeOH, što je izazvalo taloženje produkta. Talog je sakupljen filtriranjem i žuti čvrsti produkt je osušen na 80°C u vakuumu tijekom 24 h. Prinos produkta bio je 59%, a talište je bilo 135-136°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 2-Chloro-4-aminothiophenol (2.0 g) and 3,4-dichloronitrobenzene (1 eq.) were added to 300 ml of acetone containing K 2 CO 3 (20 g). The mixture was refluxed for 24 h at 60°C. After cooling to room temperature, the mixture was filtered and the acetone was removed by rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. The chloroform was then removed by rotary vacuum. The resulting residue was triturated with MeOH, which caused precipitation of the product. The precipitate was collected by filtration and the yellow solid was dried at 80°C under vacuum for 24 h. The product yield was 59%, and the melting point was 135-136°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-Amino-2-klorfenil-(2'-nitro-4'-klorfenil)-sulfid 4-Amino-2-chlorophenyl-(2'-nitro-4'-chlorophenyl)-sulfide

2-Klor-4-aminotiofenol (2,0 g) i 2,5-diklornitrobenzen (1 ekv.) dodani su u 300 m1 acetona koji sadrži K2CO3 (20 g). Smjesa je refluksirana 24 h na 60°C. Nakon hlađenja na sobnu temperaturu, smjesa je filtrirana i aceton je uklonjen rotacijskim vakuumom. Dobivene rezidue su otopljene u malom volumenu CH2Cl2 i filtrirane. The CH2Cl2 je zatim uklonjen rotacijskim vakuumom. Dobivene rezidue su razmuljene s MeOH, što je izazvalo taloženje produkta. Talog je sakupljen filtriranjem i žuti čvrsti produkt je osušen na 60°u vakuumu tijekom 24 h. Prinos produkta bio je 57%, a talište je bilo 191-193°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 2-Chloro-4-aminothiophenol (2.0 g) and 2,5-dichloronitrobenzene (1 eq.) were added to 300 ml of acetone containing K 2 CO 3 (20 g). The mixture was refluxed for 24 h at 60°C. After cooling to room temperature, the mixture was filtered and the acetone was removed by rotary vacuum. The obtained residues were dissolved in a small volume of CH2Cl2 and filtered. The CH2Cl2 was then removed by rotary vacuum. The resulting residue was triturated with MeOH, which caused precipitation of the product. The precipitate was collected by filtration and the yellow solid was dried at 60° under vacuum for 24 h. The product yield was 57%, and the melting point was 191-193°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-Amino-2-klorfenil-6-(5-nitrokinolino)-sulfid 4-Amino-2-chlorophenyl-6-(5-nitroquinolino)-sulfide

2-Klor-4-aminotiofenol (0,474 g) i 6-klor-5-nitrokinolin (1 ekv.) dodani su u 250 ml acetona koji sadrži K2CO3 (20 g). Smjesa je refluksirana 24 h na 60°C. Nakon hlađenja na sobnu temperaturu, smjesa je filtrirana i aceton je uklonjen rotacijskim vakuumom. Dobivene rezidue su otopljene u malom volumenu kloroforma i filtrirane. Zatim je uklonjen kloroform rotacijskim vakuumom. Dobivene rezidue su razmuljene s MeOH, što je izazvalo taloženje produkta. Produkt je dobiven korištenjem preparativne kromatografije primjenom otopljenih rezidua na malenu kolonu koja je napunjena silika-gelom (70-230 mesh), a zatim eluiranjem korištenjem sustava otapala kloroform:heksan (3:2). Frakcije koje sadrže produkt su spojene i otapalo je uklonjeno korištenjem rotacijskog vakuuma. Ćvrsti žuti produkt je sakupljen filtriranjem i osušen na 50°C u vakuumu tijekom 24 h. Prinos produkta bio je 62%, a talište je bilo 129-131°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 2-Chloro-4-aminothiophenol (0.474 g) and 6-chloro-5-nitroquinoline (1 eq.) were added to 250 ml of acetone containing K 2 CO 3 (20 g). The mixture was refluxed for 24 h at 60°C. After cooling to room temperature, the mixture was filtered and the acetone was removed by rotary vacuum. The obtained residues were dissolved in a small volume of chloroform and filtered. Chloroform was then removed by rotary vacuum. The resulting residue was triturated with MeOH, which caused precipitation of the product. The product was obtained using preparative chromatography by applying the dissolved residues to a small column filled with silica gel (70-230 mesh) and then elution using the solvent system chloroform:hexane (3:2). Fractions containing the product were combined and the solvent was removed using rotary vacuum. The solid yellow product was collected by filtration and dried at 50°C under vacuum for 24 h. The product yield was 62%, and the melting point was 129-131°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

D. Specifični sintetski postupci D. Specific synthetic procedures

Definirane su cjelovite metode priređivanja sljedećih spojeva: Complete methods of preparation of the following compounds are defined:

1-Acetamido-3-klor-4-(2,3-diklorfenilsulfanil)-benzen 1-Acetamido-3-chloro-4-(2,3-dichlorophenylsulfanyl)-benzene

3-Hidroksi-4-(2,3-diklorfenilsulfanil)-fenilamin hidroklorid 3-Hydroxy-4-(2,3-dichlorophenylsulfanyl)-phenylamine hydrochloride

6-Klor-5-(2,4-diklorfenilsulfanil)-1H-benzimidazol. 6-Chloro-5-(2,4-dichlorophenylsulfanyl)-1H-benzimidazole.

1-Acetamido-3-klor-4-(2,3-diklorfenillsulfanil)-benzen 1-Acetamido-3-chloro-4-(2,3-dichlorophenylsulfanyl)-benzene

0,165 g (2,1 ekv., 1,35 mmol) 4-dimetilaminopiridina (4-DMAP) stavljeno je u suhu tikvicu u atomosferi dušika i otopljeno u 5 ml bezvodnog tetrahidrofurana (THF). Dodano je 2 ml acetanhidrida, a zatim 0,220 g (1 ekv., 0,643 nmol) 3-klor-4-(2,3-diklorfenilsulfanil)-fenilamin hidroklorida. Miješano je 18 sati. Smjesa je zatim razrijeđena sa 75 ml etera i isprana zasićenom otopinom NaHCO3 (3×50 ml), 0,3 N otopinom HCl (3×30 ml) i zasićenom otopinom NaCl (2×30 ml), osušena iznad Na2SO4, filtrirana i otapalo je uklonjeno na rotacijskom uparivaču. “Flash” kromatografija (1,9×27 cm, EtAc/heptan (1:1)) dala je 0,135 g (61% prinos) bijele krutine, talište 167-169°C, Rf 0,25 (EtAc/heptan 1:1). 1H-NMR (DMSO-d6) 2.07 (s, 3H), 6.59 (d od d, J1=1.3, J2=7.9, 1H), 7.24 (t, J=8.1, 1H), 7.46 (d od d, J1=1.4, J2=8.1, 1H), 7.55 (m, 2H), 8.04 (d, J=1.9, 1H), 10.35 (s, 1H). MS (EI) m/z 345 (M+, 82). Analitički izračunato za C14H10Cl3NOS: C, 48.51; H, 2.91; N, 4.04. Nađeno: C. 48.29; H, 2.88; N, 3.92. 0.165 g (2.1 eq., 1.35 mmol) of 4-dimethylaminopyridine (4-DMAP) was placed in a dry flask under a nitrogen atmosphere and dissolved in 5 ml of anhydrous tetrahydrofuran (THF). 2 mL of acetic anhydride was added followed by 0.220 g (1 eq., 0.643 nmol) of 3-chloro-4-(2,3-dichlorophenylsulfanyl)-phenylamine hydrochloride. It's mixed at 6 p.m. The mixture was then diluted with 75 ml of ether and washed with saturated NaHCO3 solution (3×50 ml), 0.3 N HCl solution (3×30 ml) and saturated NaCl solution (2×30 ml), dried over Na2SO4, filtered and solvent was removed on the rotary evaporator. Flash chromatography (1.9×27 cm, EtAc/heptane (1:1)) gave 0.135 g (61% yield) of a white solid, mp 167-169°C, Rf 0.25 (EtAc/heptane 1: 1). 1H-NMR (DMSO-d6) 2.07 (s, 3H), 6.59 (d from d, J1=1.3, J2=7.9, 1H), 7.24 (t, J=8.1, 1H), 7.46 (d from d, J1 =1.4, J2=8.1, 1H), 7.55 (m, 2H), 8.04 (d, J=1.9, 1H), 10.35 (s, 1H). MS (EI) m/z 345 (M+, 82). Analytical calculated for C14H10Cl3NOS: C, 48.51; H, 2.91; N, 4.04. Found: C. 48.29; H, 2.88; N, 3.92.

3-Hidroksi-4-(2,3-diklorfenilsulfanil)-fenilamin hidroklorid 3-Hydroxy-4-(2,3-dichlorophenylsulfanyl)-phenylamine hydrochloride

0,19 g (0.67 mmol) 3-metoksi-4-(2,3-diklorfenilsulfanil) fenilamina stavljeno je u suhu tikvicu u atmosferi dušika, otopljeno u 10 ml suhog CH2Cl2 te ohlađeno na -78°. 0,32 ml (5 ekv., 3,3 mmol) bor-tribromida dodano je dokapavanjem, uz miješanje. Hladna kupelj je uklonjena i smjesa je ostavljena da reagira 20 sati. Otopina je ponovo ohlađena na -78°, te je dokapavanjem dodano 10 ml MeOH. Ostavljeno je da se zagrije na sobnu temperaturu i miješano je 1 sat. Otapalo je uklonjeno na rotacijskom uparivaču i dobiveno ulje je pokupljeno s 10 ml MeOH i opet je uklonjeno otapalo. Tvar je opet pokupljena s 10 ml MeOH i zatim je razrijeđena s 100 ml EtAc. Bijeli talog je uklonjen filtriranjem, pa je filtrat ispran zasićenom otopinom NaCl (3×50 ml), osušen iznad Na2SO4, filtriran, pa je otapalo uklonjeno rotacijskim uparivačem. Dobiveno ulje je pročišćeno “flash” kromatografijom (2,9×28 cm, EtAc/heptan (1:3)) pa je otopljeno u 25 ml etera i staloženo kao HCl sol dodatkom 5 ml 1 N HCl u eteru. Talog je sakupljen filtriranjem, ispran eterom, pa osušen (90ºC, 3 h, 0,3 mm Hg) da se dobije 0,17 g (80% prinos) bijele krutine, talište 222°C (raspad), Rf 0,49 (EtAc/heptan 1:1). 1H-NMR (DMSO-d6) 6.56-6.61 (m, 2H), 6.79 (d, J=1.9, 1H), 6.90 (br hump), 7.19 (t, J=8.1, 1H), 7.27 (d, J=8.2, 1H), 7.38 (d od d, J1=1.4, J2=8.1, 1H), 10.37 (br s, 1H). MS (EI) m/z 285 M+ [100]. Analitički izračunato za C12H9Cl2NOS·HCl: C, 44.67; H, 3.12; N, 4.34. Nađeno: C, 44.53; H, 2.91; N, 4.17. 0.19 g (0.67 mmol) of 3-methoxy-4-(2,3-dichlorophenylsulfanyl)phenylamine was placed in a dry flask in a nitrogen atmosphere, dissolved in 10 ml of dry CH2Cl2 and cooled to -78°. 0.32 ml (5 eq., 3.3 mmol) of boron tribromide was added dropwise with stirring. The cold bath was removed and the mixture was allowed to react for 20 hours. The solution was cooled again to -78°, and 10 ml of MeOH was added dropwise. It was allowed to warm to room temperature and stirred for 1 hour. The solvent was removed on a rotary evaporator and the resulting oil was taken up with 10 ml of MeOH and the solvent was again removed. The material was taken up again with 10 ml of MeOH and then diluted with 100 ml of EtAc. The white precipitate was removed by filtration, and the filtrate was washed with saturated NaCl solution (3×50 ml), dried over Na2SO4, filtered, and the solvent was removed using a rotary evaporator. The obtained oil was purified by "flash" chromatography (2.9×28 cm, EtAc/heptane (1:3)), then it was dissolved in 25 ml of ether and settled as an HCl salt by adding 5 ml of 1 N HCl in ether. The precipitate was collected by filtration, washed with ether, then dried (90ºC, 3 h, 0.3 mm Hg) to give 0.17 g (80% yield) of a white solid, mp 222°C (dec), Rf 0.49 ( EtAc/heptane 1:1). 1H-NMR (DMSO-d6) 6.56-6.61 (m, 2H), 6.79 (d, J=1.9, 1H), 6.90 (br hump), 7.19 (t, J=8.1, 1H), 7.27 (d, J =8.2, 1H), 7.38 (d of d, J1=1.4, J2=8.1, 1H), 10.37 (br s, 1H). MS (EI) m/z 285 M+ [100]. Analytical calculated for C12H9Cl2NOS·HCl: C, 44.67; H, 3.12; N, 4.34. Found: C, 44.53; H, 2.91; N, 4.17.

6-Klor-5-(2,4-diklorfenilsulfanil-1H-benzimidazol 6-Chloro-5-(2,4-dichlorophenylsulfanyl-1H-benzimidazole).

4-Klor-2-nitro-5-(2,4-diklorfenilsulfanil)-fenilamin priređen je općim postupkom s izuzetkom da je pročišćen kao slobodan amin “flash” kromatografijom. 1,37 g (3,93 mmol) slobodonog amina je dodano u 10 ml DMF i 10 ml EtOH. 4,43 g (5 ekv., 19,7 mmol) kositrenog klorida dihidrata je dodano uz 10 ml koncentrirane HCl. Grijano je na 60°C tijekom 20 sati. Reakcijska smjesa je ostavljena da se ohladi na sobnu temperaturu, zatim je razrijeđena s 30 ml vode, te dovedena na pH 12 dodatkom 30 ml 5 N NaOH. Smjesa je zatim dva puta ekstrahirana sa 150 ml etera. Sjedinjeni organski slojevi isprani sa sa 100 ml zasićene otopine NaHCO3 i 2×100 ml zasićene otopine NaCl, osušeni iznad Na2SO4 i filtrirani. Dodano je 40 ml heptana, te je koncentrirano na rotacijskom uparivaču i osušeno u vakuumu (100°C, 2 sata, 0,3 mm Hg) da se dobije 1,06 g (82%) analitički čiste bijele krutine, talište 206-208°C, Rf 0,51 (CH2Cl2/MeOH (9:1) s 1% TEA). 1H-NMR (DMSO-d6) 6.63 (d, J=8.6, 1H), 7.28 (d od d, J1=2.2, J2=8.7, 1H), 7.69 (d, J=2.2; 1H), 7.86 (br s, 1H), 7.93 (s, 1H), 8.38 (s, 1H), 12.80 (s, 1H). MS (EI) m/z 328 (M+, 97). Analitički izračunato za C13H7Cl3N2S: C, 47.37; H, 2.14; N, 8.50. Nađeno: C, 47.40; H, 2.04; N, 8.32. 4-Chloro-2-nitro-5-(2,4-dichlorophenylsulfanyl)-phenylamine was prepared by the general procedure except that it was purified as the free amine by flash chromatography. 1.37 g (3.93 mmol) of the free amine was added to 10 ml of DMF and 10 ml of EtOH. 4.43 g (5 eq., 19.7 mmol) of stannous chloride dihydrate was added with 10 ml of concentrated HCl. It was heated at 60°C for 20 hours. The reaction mixture was allowed to cool to room temperature, then diluted with 30 ml of water, and brought to pH 12 by adding 30 ml of 5 N NaOH. The mixture was then extracted twice with 150 ml of ether. The combined organic layers were washed with 100 ml saturated NaHCO3 solution and 2×100 ml saturated NaCl solution, dried over Na2SO4 and filtered. 40 mL of heptane was added and concentrated on a rotary evaporator and dried in vacuo (100°C, 2 hours, 0.3 mm Hg) to give 1.06 g (82%) of an analytically pure white solid, mp 206-208 °C, Rf 0.51 (CH 2 Cl 2 /MeOH (9:1) with 1% TEA). 1H-NMR (DMSO-d6) 6.63 (d, J=8.6, 1H), 7.28 (d from d, J1=2.2, J2=8.7, 1H), 7.69 (d, J=2.2; 1H), 7.86 (br s, 1H), 7.93 (s, 1H), 8.38 (s, 1H), 12.80 (s, 1H). MS (EI) m/z 328 (M+, 97). Analytical calculated for C13H7Cl3N2S: C, 47.37; H, 2.14; N, 8.50. Found: C, 47.40; H, 2.04; N, 8.32.

E. Specifični protokoli sinteze E. Specific synthesis protocols

Sljedeći spojevi priređeni su metodama koje su niže opisane: The following compounds were prepared by the methods described below:

4-Metilamino-2,2',4'-triklordifenilsulfid 4-Methylamino-2,2',4'-trichlorodiphenylsulfide

4-Amino-2-klorfenil-(4'-acetamido-2'-klorfenil)-sulfid 4-Amino-2-chlorophenyl-(4'-acetamido-2'-chlorophenyl)-sulfide

4-Amino-2-klorfenil-(4'-dimetilamino-2'-klorfenil)-sulfid 4-Amino-2-chlorophenyl-(4'-dimethylamino-2'-chlorophenyl)-sulfide

4-Aminometil-2-klorfenil-(2',4'-diklorfenil)-sulfid 4-Aminomethyl-2-chlorophenyl-(2',4'-dichlorophenyl)-sulfide

4-Metilamino-2,2',4'-triklordifenilsulfid 4-Methylamino-2,2',4'-trichlorodiphenylsulfide

4-Amino-2,2',4'-triklordifenil-sulfid (0,305 g, 1,0 mmol) dodan je u 15 ml mravlje kiseline. Smjesa je zatim miješana 8 sati, nakon čega je dodano 0,23 ml 37% formaldehida i smjesa je refluksirana 8 sati. Otapalo je aztim uklonjeno na rotacijskom uparivaču. Dobivene rezidue su primijenjene na malenu kolonu koja sadrži silika-gel (70-230 mesh). Produkt je zatim eluiran s kolone kloroformom. Otapalo je uklonjeno iz eluata na rotacijskom uparivaču i sakupljen je blijedožuti produkt. Prinos produkta je bio 44%, a talište je bilo 211°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 4-Amino-2,2',4'-trichlorodiphenyl-sulfide (0.305 g, 1.0 mmol) was added to 15 ml of formic acid. The mixture was then stirred for 8 hours, after which 0.23 ml of 37% formaldehyde was added and the mixture was refluxed for 8 hours. The solvent was removed on a rotary evaporator. The obtained residues were applied to a small column containing silica gel (70-230 mesh). The product was then eluted from the column with chloroform. The solvent was removed from the eluate on a rotary evaporator and the pale yellow product was collected. The product yield was 44%, and the melting point was 211°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-Nitro-2-klorfenil-(4'-acetamido-2'-klorfenil)-sulfid 4-Nitro-2-chlorophenyl-(4'-acetamido-2'-chlorophenyl)-sulfide

4-Amino-2,2'-diklor-4'-nitrodifenilsulfid (1,0 g, 3,17 mmol) zagrijan je u 15 ml acetanhidrida koji sadrži p-toluenesulfonske kisleine. Smjesa je ostavljena stajati 1 sat, nakon čega je uklonjeno otapalo na rotacijskom uparivaču. Rezidue su otopljene u EtAc i stavljene na malenu kolonu silika-gela (70-230 mesh). Filtrat je sakupljen, uparen do suhoga i rekristaliziran iz acetonitrila da se dobije žuti čvrsti produkt. Prinos produkta bio je 97%, a talište je bilo 163-165°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 4-Amino-2,2'-dichloro-4'-nitrodiphenylsulfide (1.0 g, 3.17 mmol) was heated in 15 ml of acetic anhydride containing p-toluenesulfonic acids. The mixture was allowed to stand for 1 hour, after which the solvent was removed on a rotary evaporator. The residues were dissolved in EtAc and applied to a small column of silica gel (70-230 mesh). The filtrate was collected, evaporated to dryness and recrystallized from acetonitrile to give a yellow solid. The product yield was 97%, and the melting point was 163-165°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-Nitro-2-klorfenil-(4'-dimetilamino-2'-klorfenil)-sulfid 4-Nitro-2-chlorophenyl-(4'-dimethylamino-2'-chlorophenyl)-sulfide

4-Amino-2-klorfenil-(2'-klor-4'-nitrofenil)-sulfid (1,0 g, 3,17 mmol)dodan je u suspenziju 60% natrijeva hidrida (1,43 g) u 250 ml THF na 0°C. Zatim je dodan jodmetan (0,2 ml, u 20 m1 THF), te je smjesa miješana na sobnoj temperaturi 48 sati. Smjesa je zatim primijenjena na Analtech silika-gel ploče. Svaka ploča je bila debljine 1000 mikrona silika-gela. Dva reakcijska produkta, tj. 4-nitro-2-klorfenil-(4'-metilamino-2'-klorfenil)-sulfid i 4-nitro-2-klorfenil-(4'-dimetilamino-2'-klorfenil)-sulfid, eluirani su smjesom 1:1 CHCl3 i heksana. Sporija elucijska vrpca odgovara prvom produktu, a brža elucijska vrpca drugom produktu. Nakon sakupljanja vrpci, spojevi su otopljeni u CHCl3. Otapalo je uklonjeno rotacijskim uparavanjem da se dobije žuti čvrsti produkt. Prinos željenog produkta bio je 36%, a talište je bilo 138-139°C. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 4-Amino-2-chlorophenyl-(2'-chloro-4'-nitrophenyl)sulfide (1.0 g, 3.17 mmol) was added to a suspension of 60% sodium hydride (1.43 g) in 250 ml THF at 0°C. Then iodomethane (0.2 ml, in 20 ml of THF) was added, and the mixture was stirred at room temperature for 48 hours. The mixture was then applied to Analtech silica-gel plates. Each plate was 1000 micron thick silica gel. Two reaction products, i.e. 4-nitro-2-chlorophenyl-(4'-methylamino-2'-chlorophenyl)-sulfide and 4-nitro-2-chlorophenyl-(4'-dimethylamino-2'-chlorophenyl)-sulfide, they were eluted with a 1:1 mixture of CHCl3 and hexane. A slower elution band corresponds to the first product, and a faster elution band to the second product. After collecting the bands, the compounds were dissolved in CHCl3. The solvent was removed by rotary evaporation to give a yellow solid. The yield of the desired product was 36%, and the melting point was 138-139°C. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

4-Aminometil-2-klorfenil-(2',4'-diklorfenil)-sulfid 4-Aminomethyl-2-chlorophenyl-(2',4'-dichlorophenyl)-sulfide

2-Klor-4-cijanofenil-(2',4'-diklorfenil)-sulfid (1,0 g, 3,18 mmol) dodan je u 200 ml THF pod dušikom na 0°C. Zatim je u otopinu u obrocima dodan litij-aluminijev hidrid (0,24 g). Smjesa je zatim ostavljena stajati 2 sata na 0°C, nakon čega je u smjesu dodan 1 ml 20% otopine NaOH, a zatim 1 ml dH2O. Otopina je zatim filtrirana, pa je otapalo uklonjeno na rotacijskom uparivaču. Dobivene rezidue su primijenjene na malenu kolonu koja sadrži silika-gel (70-230 mesh). Kolona je isprana s 1:1 smjesom kloroforma i heksana, pa je produkt eluiranjem korištenjem rotacijskog uparivača, a produkt je dobiven kao bezbojno ulje. Prinos produkta bio je 40%, a talište nije određeno. Za karakteriziranje spoja korištene su standardne analitičke tehnike, uključujući NMR spektroskopiju, elementarnu analizu i tankoslojnu kromatografiju. 2-Chloro-4-cyanophenyl-(2',4'-dichlorophenyl)sulfide (1.0 g, 3.18 mmol) was added to 200 mL of THF under nitrogen at 0°C. Lithium aluminum hydride (0.24 g) was then added to the solution in portions. The mixture was then left to stand for 2 hours at 0°C, after which 1 ml of 20% NaOH solution was added to the mixture, followed by 1 ml of dH2O. The solution was then filtered, and the solvent was removed on a rotary evaporator. The obtained residues were applied to a small column containing silica gel (70-230 mesh). The column was washed with a 1:1 mixture of chloroform and hexane, and the product was eluted using a rotary evaporator, and the product was obtained as a colorless oil. The yield of the product was 40%, and the melting point was not determined. Standard analytical techniques, including NMR spectroscopy, elemental analysis and thin-layer chromatography, were used to characterize the compound.

F. Ostali spojevi F. Other compounds

Sukladno općoj metodi sinteze sintetizirani su sljedeći spojevi: According to the general method of synthesis, the following compounds were synthesized:

Tablica II Table II

4-Nitro-2-klorfenil-(2',4'-dimetilfenil)-sulfid 4-Nitro-2-chlorophenyl-(2',4'-dimethylphenyl)-sulfide

4-Nitro-2-klorfenil-(2'-metil-4'-klorfenil)-sulfid 4-Nitro-2-chlorophenyl-(2'-methyl-4'-chlorophenyl)-sulfide

4-Nitro-2-klorfenil-(2', 4'-difluorfenil)-sulfid 4-Nitro-2-chlorophenyl-(2', 4'-difluorophenyl)-sulfide

4-Nitro-2-klorfenil-(2', 4', 6'-triklorfenil)-sulfid 4-Nitro-2-chlorophenyl-(2', 4', 6'-trichlorophenyl)-sulfide

4-Nitro-2-klorfenil-(3', 4'-diklorfenil)-sulfid 4-Nitro-2-chlorophenyl-(3', 4'-dichlorophenyl)-sulfide

4-Nitro-2-klorfenil-2-(3-klor-5-trifluormetilpiridil)-sulfid 4-Nitro-2-chlorophenyl-2-(3-chloro-5-trifluoromethylpyridyl)-sulfide

2-Klor-4-nitrofenil-(2', 4'-diklorfenil)-sulfid 2-Chloro-4-nitrophenyl-(2', 4'-dichlorophenyl)-sulfide

4-Nitro-2-klorfenil-(4'-acetamido-2'-klorfenil)-sulfid 4-Nitro-2-chlorophenyl-(4'-acetamido-2'-chlorophenyl)-sulfide

4-Nitro-2-klorfenil-(4'-dimetilamino-2'-klorfenil)-sulfid 4-Nitro-2-chlorophenyl-(4'-dimethylamino-2'-chlorophenyl)-sulfide

2-Klor-4-nitro-5-metilaminofenil-(2', 4'-dich1orofenil)-sulfid 2-Chloro-4-nitro-5-methylaminophenyl-(2', 4'-dichlorophenyl)-sulfide

2-Klor-4-nitro-5-morfolinofenil-(2', 4'-diklorfenil)-sulfid 2-Chloro-4-nitro-5-morpholinophenyl-(2', 4'-dichlorophenyl)-sulfide

4-Nitro-2-trifluormetilfenil-(2', 4'-diklorfenil)-sulfid 4-Nitro-2-trifluoromethylphenyl-(2', 4'-dichlorophenyl)-sulfide

4-Nitro-2-fluorfenil-(2', 4'-diklorfenil)-sulfid 4-Nitro-2-fluorophenyl-(2', 4'-dichlorophenyl)-sulfide

3-Klor-5-nitrofenil-(2', 4'-diklorfenil)-sulfid 3-Chloro-5-nitrophenyl-(2', 4'-dichlorophenyl)-sulfide

4-Nitro-2-klorfenil-( 1-naftil)-sulfid 4-Nitro-2-chlorophenyl-(1-naphthyl)-sulfide

4-Nitro-2-metilfenil-(2',4'-diklorfenil)-sulfid 4-Nitro-2-methylphenyl-(2',4'-dichlorophenyl)-sulfide

4-Nitro-2-bromfenil-(2',4'-diklorfenil)-sulfid 4-Nitro-2-bromophenyl-(2',4'-dichlorophenyl)-sulfide

4-Nitro-2, 5-diklorfenil-(2',4'-diklorfenil)-sulfid 4-Nitro-2, 5-dichlorophenyl-(2',4'-dichlorophenyl)-sulfide

4-Nitro-2, 6-diklorfenil-(2',4'-diklorfenil)-sulfid 4-Nitro-2, 6-dichlorophenyl-(2',4'-dichlorophenyl)-sulfide

4-Nitro-2-klor-5-fluorfenil-(2', 4'-diklorfenil)-sulfid 4-Nitro-2-chloro-5-fluorophenyl-(2', 4'-dichlorophenyl)-sulfide

4-Nitro-2, 3-diklorfenil-(2', 4'-diklorfenil)-sulfid 4-Nitro-2, 3-dichlorophenyl-(2', 4'-dichlorophenyl)-sulfide

4-Nitro-2-klorfenil-(4'-klor-2'-aminofenil)-sulfid 4-Nitro-2-chlorophenyl-(4'-chloro-2'-aminophenyl)-sulfide

4-Nitro-5-acetamido-2-klorfenil-(2', 4'-diklorfenil)-sulfid 4-Nitro-5-acetamido-2-chlorophenyl-(2', 4'-dichlorophenyl)-sulfide

4-Nitro-2-klorfenil-(4'-metilamino-2'-klorfenil)-sulfid 4-Nitro-2-chlorophenyl-(4'-methylamino-2'-chlorophenyl)-sulfide

4-Nitro-2-klorfenil-(4'-benzilamino-2'-klorfenil)-sulfid 4-Nitro-2-chlorophenyl-(4'-benzylamino-2'-chlorophenyl)-sulfide

4-Nitro-2-klorfenil-(4'-dibenzilamino-2'-klorfenil)-sulfid 4-Nitro-2-chlorophenyl-(4'-dibenzylamino-2'-chlorophenyl)-sulfide

4-Nitro-5-fenilsulfo-2-klorfenil-(2', 4'-diklorfenil)-sulfid 4-Nitro-5-phenylsulfo-2-chlorophenyl-(2', 4'-dichlorophenyl)-sulfide

3-Nitro-5-klorfenil-(2', 4'-diklorfenil)-sulfid 3-Nitro-5-chlorophenyl-(2', 4'-dichlorophenyl)-sulfide

Primjer 4 Example 4

Rezultati stanične analize Results of cellular analysis

Spojevi ovog izuma pokazali su se aktivnima u staničnim analizama koje su prije opisane, te su navedeni u tablici III niže, u kojoj su navedene µM IC50 vrijednosti inhibiranja LFA-1 vezanja za ICAM-1 i ICAM-3 koje su ispitane. Sparene vrijednosti (X/Y) označuju inhibiranje u odsutnosti i prisutnosti IL-8. Višestruke sparene vrijednosti (W/X; Y/Z) označuju ponovljene eksperimente. Crtice (--,X) označuju da eksperiment nije izvršen. “NT” označuje da spoj nije ispitan u konkretnoj analizi. The compounds of this invention were shown to be active in the cellular assays previously described, and are listed in Table III below, which lists the µM IC50 values of inhibition of LFA-1 binding to ICAM-1 and ICAM-3 that were tested. Paired values (X/Y) indicate inhibition in the absence and presence of IL-8. Multiple paired values (W/X; Y/Z) indicate repeated experiments. Dashes (--,X) indicate that the experiment was not performed. "NT" indicates that the compound was not tested in the specific analysis.

Mada je ovaj izum prikazan specifičnim realizacijama, podrazumijeva se da su varijacije i modifikacije moguće za one koji poznaju ovo pdoručje. Prema tome, samo ona ograničenja poput onih u priloženim patentnim zahtjevima mogu se odnositi na ovaj izum. Although this invention is shown in specific embodiments, it is understood that variations and modifications are possible to those skilled in the art. Accordingly, only such limitations as those in the appended claims may apply to this invention.

Tablica III Table III

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Claims (20)

1. Spoj, naznačen time što je odabran iz skupa kojega sačinjavaju: 3-Klor-4-(2-klorfenilsulfanil)-fenilamin hidroklorid 3-Klor-4-(2-naftilsulfanil)-fenilamin hidroklorid 3-Klor-4-(2,3-diklorfenilsulfanil)-fenilamin hidroklorid 3-Klor-4-(2,4,5-triklorfenilsulfanil)-fenilamin hidroklorid 3-Klor-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 3-Metoksi-4-(2,3-diklorfenilsulfanil)-fenilamin 5-Amino-2-(2,3-diklorfenilsulfanil)-acetofenon hidroklorid 4-(2,3-diklorfenilsulfanil)-fenilamin 3-Klor-4-(1-naftilsulfanil)-fenilamin hidroklorid 3-Metil-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 1-Acetamido-3-klor-4-(2,3-diklorfenilsulfanil)-benzen 4-Metilamino-2,2',4'-triklordifenilsulfid 3-Brom-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 3-Hidroksi-4-(2,3-diklorfenilsulfanil)-fenilamin hidroklorid 6-Klor-5-(2,4-diklorfenilsulfanil)-1 H-benzimidazole 4-Amino-2-klorfenil-(2'4'-dimetilfenil)-sulfid hidroklorid 2,5-Diklor-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 4-Amino-2-klorfenil-(2'-metil-4'-klorfenil)-sulfid 4-Amino-2-klorfenil-(2',4'-difluorfenil)-sulfid hidroklorid 4-Amino-2-klorfenil-(2',4',6'-triklorfenil)-sulfid 4-Amino-2-klorfenil-(2'-amino-4'-klorfenil)-sulfid 4-Amino-2-klorfenil-(2'-klor-4'-nitrofenil)-sulfid 4-Amino-2-klorfenil-(2'-nitro-4'-klorfenil)-sulfid 4-Amino-2-klorfenil-(3',4'-diklorfenil)-sulfid 4-Amino-2-klorfenil-2-(3-klor-5-trifluormetilpiridil)-sulfid Bis-(4,4'-diamino-2,2'-diklorfenil)-sulfid 4-Amino-2-klorfenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-klorfenil-(4'-acetamido-2'-klorfenil)-sulfid 4-Amino-2-klorfenil-6-(5-nitrokinolino)-sulfid 4-Amino-2-klorfenil-(4'-dimetilamino-2'-klorfenil)-sulfid 2-Klor-4-amino-5-metilaminofenil-(2',4'-diklorfenil)-sulfid 2-Klor-4-amino-5-N-morfolinofenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-trifluormetilfenil-(2',4'-diklorfenil)-sulfid 4-Aminometil-2-klorfenil-(2', 4'-diklorfenil)-sulfid 4,5-Diklor-2-(2,4-diklorfenilsulfanil)-fenilamin 3,5-Diklor-4-(2,4-diklorfenilsulfanil)-fenilamin 2,3-Diklor-4-(2,4-diklorfenilsulfanil)-fenilamin 4-Amino-2-fluorfenil-(2',4'-diklorfenil)-sulfid 5-Amino-3-klorfenil-(2',4'-diklorfenil)-sulfid 4-Nitro-2-klorfenil-(2’,4’-dimetilfenil)-sulfid 4-Nitro-2-klorfenil-(2’-metil-4’-klorfenil)-sulfid 4-Nitro-2-klorfenil-(2’,4’-difluorfenil)-sulfid 4-Nitro-2-klorfenil-(2’,4’,6’-triklorfenil)-sulfid 4-Nitro-2-klorfenil-(3’,4’-diklorfenil)-sulfid 4-Nitro-2-klorfenil-2-(3-klor-5-trifluormetilpiridil)-sulfid 2-Klor-4-nitrofenil-(2’,4’-diklorfenil)-sulfid 4-Nitro-2-klorfenil-(4’-acetamido-2’-klorfenil)sulfid 4-Nitro-2-klorfenil-(4’-dimetilamino-2’-klorfenil)sulfid 2-Klor-4-nitro-5-metilaminofenil-(2’,4’-diklorfenil)-sulfid 2-Klor-4-nitro-5-morfolinofenil-(2’,4’-diklorfenil)-sulfid 4-Nitro-2-trifluormetilfenil-(2’,4’-diklorfenil)-sulfid 4-Nitro-2-fluorfenil-(2’,4’-diklorfenil)-sulfid 3-Klor-5-nitrofenil-(2’,4’-diklorfenil)-sulfid 4-Nitro-2-klorfenil-(1-naftil)-sulfid 4-Nitro-2-metilfenil-(2’,4’-diklorfenil)-sulfid 4-Nitro-2-bromfenil-(2’,4’-diklorfenil)-sulfid 4-Nitro-2,5-diklorfenil-(2’,4’-diklorfenil)-sulfid 4-Nitro-2,6-diklorfenil-(2’,4’-diklorfenil)-sulfid 4-Nitro-2-klor-5-fluorfenil-(2’,4’-diklorfenil)-sulfid 4-Nitro-2,3-diklorfenil-(2’,4’-diklorfenil)-sulfid 4-Nitro-2-klorfenil-(4’-klor-2’-aminofenil)-sulfid 4-Nitro-5-acetamido-2-klorfenil-(2’,4’-diklorfenil)-aulfid 4-Nitro-2-klorfenil-(4’-metilamino-2’-klorfenil)-sulfid 4-Nitro-2-klorfenil-(4’-benzilamino-2’-klorfenil)-sulfid 4-Nitro-2-klorfenil-(4’-dibenzilamino-2’-klorfenil)-sulfid 4-Nitro-5-fenilsulfo-2-klorfenil-(2’,4’-diklorfenil)-sulfid 3-Nitro-5-klorfenil-(2’,4’-diklorfenil)-sulfid1. Compound, indicated by the fact that it is selected from the set consisting of: 3-Chloro-4-(2-chlorophenylsulfanyl)-phenylamine hydrochloride 3-Chloro-4-(2-naphthylsulfanyl)-phenylamine hydrochloride 3-Chloro-4-(2,3-dichlorophenylsulfanyl)-phenylamine hydrochloride 3-Chloro-4-(2,4,5-trichlorophenylsulfanyl)-phenylamine hydrochloride 3-Chloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride 3-Methoxy-4-(2,3-dichlorophenylsulfanyl)-phenylamine 5-Amino-2-(2,3-dichlorophenylsulfanyl)-acetophenone hydrochloride 4-(2,3-dichlorophenylsulfanyl)-phenylamine 3-Chloro-4-(1-naphthylsulfanyl)-phenylamine hydrochloride 3-Methyl-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride 1-Acetamido-3-chloro-4-(2,3-dichlorophenylsulfanyl)-benzene 4-Methylamino-2,2',4'-trichlorodiphenylsulfide 3-Bromo-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride 3-Hydroxy-4-(2,3-dichlorophenylsulfanyl)-phenylamine hydrochloride 6-Chloro-5-(2,4-dichlorophenylsulfanyl)-1H-benzimidazoles 4-Amino-2-chlorophenyl-(2'4'-dimethylphenyl)-sulfide hydrochloride 2,5-Dichloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride 4-Amino-2-chlorophenyl-(2'-methyl-4'-chlorophenyl)-sulfide 4-Amino-2-chlorophenyl-(2',4'-difluorophenyl)-sulfide hydrochloride 4-Amino-2-chlorophenyl-(2',4',6'-trichlorophenyl)sulfide 4-Amino-2-chlorophenyl-(2'-amino-4'-chlorophenyl)-sulfide 4-Amino-2-chlorophenyl-(2'-chloro-4'-nitrophenyl)-sulfide 4-Amino-2-chlorophenyl-(2'-nitro-4'-chlorophenyl)-sulfide 4-Amino-2-chlorophenyl-(3',4'-dichlorophenyl)-sulfide 4-Amino-2-chlorophenyl-2-(3-chloro-5-trifluoromethylpyridyl)-sulfide Bis-(4,4'-diamino-2,2'-dichlorophenyl)-sulfide 4-Amino-2-chlorophenyl-(2',4'-dichlorophenyl)-sulfide 4-Amino-2-chlorophenyl-(4'-acetamido-2'-chlorophenyl)-sulfide 4-Amino-2-chlorophenyl-6-(5-nitroquinolino)-sulfide 4-Amino-2-chlorophenyl-(4'-dimethylamino-2'-chlorophenyl)-sulfide 2-Chloro-4-amino-5-methylaminophenyl-(2',4'-dichlorophenyl)-sulfide 2-Chloro-4-amino-5-N-morpholinophenyl-(2',4'-dichlorophenyl)-sulfide 4-Amino-2-trifluoromethylphenyl-(2',4'-dichlorophenyl)-sulfide 4-Aminomethyl-2-chlorophenyl-(2', 4'-dichlorophenyl)-sulfide 4,5-Dichloro-2-(2,4-dichlorophenylsulfanyl)-phenylamine 3,5-Dichloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine 2,3-Dichloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine 4-Amino-2-fluorophenyl-(2',4'-dichlorophenyl)-sulfide 5-Amino-3-chlorophenyl-(2',4'-dichlorophenyl)-sulfide 4-Nitro-2-chlorophenyl-(2',4'-dimethylphenyl)-sulfide 4-Nitro-2-chlorophenyl-(2'-methyl-4'-chlorophenyl)-sulfide 4-Nitro-2-chlorophenyl-(2',4'-difluorophenyl)-sulfide 4-Nitro-2-chlorophenyl-(2',4',6'-trichlorophenyl)-sulfide 4-Nitro-2-chlorophenyl-(3',4'-dichlorophenyl)-sulfide 4-Nitro-2-chlorophenyl-2-(3-chloro-5-trifluoromethylpyridyl)-sulfide 2-Chloro-4-nitrophenyl-(2',4'-dichlorophenyl)-sulfide 4-Nitro-2-chlorophenyl-(4'-acetamido-2'-chlorophenyl)sulfide 4-Nitro-2-chlorophenyl-(4'-dimethylamino-2'-chlorophenyl)sulfide 2-Chloro-4-nitro-5-methylaminophenyl-(2',4'-dichlorophenyl)-sulfide 2-Chloro-4-nitro-5-morpholinophenyl-(2',4'-dichlorophenyl)-sulfide 4-Nitro-2-trifluoromethylphenyl-(2',4'-dichlorophenyl)-sulfide 4-Nitro-2-fluorophenyl-(2',4'-dichlorophenyl)-sulfide 3-Chloro-5-nitrophenyl-(2',4'-dichlorophenyl)-sulfide 4-Nitro-2-chlorophenyl-(1-naphthyl)-sulfide 4-Nitro-2-methylphenyl-(2',4'-dichlorophenyl)-sulfide 4-Nitro-2-bromophenyl-(2',4'-dichlorophenyl)-sulfide 4-Nitro-2,5-dichlorophenyl-(2',4'-dichlorophenyl)-sulfide 4-Nitro-2,6-dichlorophenyl-(2',4'-dichlorophenyl)-sulfide 4-Nitro-2-chloro-5-fluorophenyl-(2',4'-dichlorophenyl)-sulfide 4-Nitro-2,3-dichlorophenyl-(2',4'-dichlorophenyl)-sulfide 4-Nitro-2-chlorophenyl-(4'-chloro-2'-aminophenyl)-sulfide 4-Nitro-5-acetamido-2-chlorophenyl-(2',4'-dichlorophenyl)-sulfide 4-Nitro-2-chlorophenyl-(4'-methylamino-2'-chlorophenyl)-sulfide 4-Nitro-2-chlorophenyl-(4'-benzylamino-2'-chlorophenyl)-sulfide 4-Nitro-2-chlorophenyl-(4'-dibenzylamino-2'-chlorophenyl)-sulfide 4-Nitro-5-phenylsulfo-2-chlorophenyl-(2',4'-dichlorophenyl)-sulfide 3-Nitro-5-chlorophenyl-(2',4'-dichlorophenyl)-sulfide 2. Farmaceutska smjesa, naznačena time što sadrži spoj prema zahtjevu 1 i farmaceutski prihvatljivi nosač.2. Pharmaceutical mixture, characterized in that it contains the compound according to claim 1 and a pharmaceutically acceptable carrier. 3. Farmaceutska smjesa, naznačena time što sadrži spoj koji je odabran iz skzpa kojega sačinjavaju: 3-Klor-4-(1-klor-naftalen-2-ilsulfanil)-fenilamin 1-(3-Nitro-4-fenilsulfanil-fenil)-etanon 1-(3-Nitro-4-fenilsulfanil-fenil)-etanon oksim 5-Trifluormetil-2-fenilsulfanil-benzonitril 1-(3,5-diklorfenil)-fenilsulfanil-pirolidin-2,5-dion Bis-2,4,6-Trinitrofenil-sulfid 2-Metil-1-(2-o-tolilsulfanil-fenil)-1H-pirol 3-[2-(4-Klor-2-nitro-fenilsulfanil)-fenilamino-3H-izobenzofuran-1-on 4-(Benzotiazol-2-ilsulfanil)-3-klor-fenilamin 2-Nitro-4-klorfenil-(2’-aminofenil)-sulfid 6-Amino-2-klorfenil-(4’-metilfenil)-sulfid 4-Nitrofenil-(2’-klorfenil)-sulfid 2,4-Dinitrofenil-(4’-klorfenil)-sulfid 4-Aminofenil-(2’-klorfenil)-sulfid 2,4-Diaminofenil)-(4’-izopropilfenil)-sulfid 4-Nitro-2-klorfenil-(2’,3’-diklorfenil)-sulfid 4-Amino-2-klorfenil-2-(5-nitro-3-brom)-piridin sulfid 4-Amino-2-klorfenil-(2’-nitro-4’-klorfenil)-sulfid3. Pharmaceutical mixture, characterized by the fact that it contains a compound selected from the group consisting of: 3-Chloro-4-(1-chloro-naphthalen-2-ylsulfanyl)-phenylamine 1-(3-Nitro-4-phenylsulfanyl-phenyl)-ethanone 1-(3-Nitro-4-phenylsulfanyl-phenyl)-ethanone oxime 5-Trifluoromethyl-2-phenylsulfanyl-benzonitrile 1-(3,5-dichlorophenyl)-phenylsulfanyl-pyrrolidine-2,5-dione Bis-2,4,6-Trinitrophenyl-sulphide 2-Methyl-1-(2-o-tolylsulfanyl-phenyl)-1H-pyrrole 3-[2-(4-Chloro-2-nitro-phenylsulfanyl)-phenylamino-3H-isobenzofuran-1-one 4-(Benzothiazol-2-ylsulfanyl)-3-chloro-phenylamine 2-Nitro-4-chlorophenyl-(2'-aminophenyl)-sulfide 6-Amino-2-chlorophenyl-(4'-methylphenyl)-sulfide 4-Nitrophenyl-(2'-chlorophenyl)-sulfide 2,4-Dinitrophenyl-(4'-chlorophenyl)-sulfide 4-Aminophenyl-(2'-chlorophenyl)-sulfide 2,4-Diaminophenyl)-(4'-isopropylphenyl)-sulfide 4-Nitro-2-chlorophenyl-(2',3'-dichlorophenyl)-sulfide 4-Amino-2-chlorophenyl-2-(5-nitro-3-bromo)-pyridine sulfide 4-Amino-2-chlorophenyl-(2'-nitro-4'-chlorophenyl)-sulfide 4. Metoda za tretiranje upalne bolesti, naznačena time što obuhvaća stupanj primjene sisavcu određene količine farmaceutske smjese prema zahtjevu 2 ili 3, koja je dovoljna za inhibiranje vezanja LFA-1 za njegov prirodni ligand koja je u kompeticiji s ICAM-1 ili ICAM-3 za vezanje s LFA-1.4. A method for treating an inflammatory disease, characterized by the step of administering to a mammal a certain amount of a pharmaceutical mixture according to claim 2 or 3, which is sufficient to inhibit the binding of LFA-1 to its natural ligand, which is in competition with ICAM-1 or ICAM-3 for binding with LFA-1. 5. Metoda za inhibiranje LFA-1 vezanje sa ICAM koji veže LFA-1, naznačena time što uključuje dovođenje u dodir LFA-1 s diaril sulfidom.5. A method for inhibiting LFA-1 binding to ICAM that binds LFA-1, comprising contacting LFA-1 with diaryl sulfide. 6. Metoda prema zahtjevu 5, naznačena time što je diaril sulfid supstituiran.6. The method according to claim 5, characterized in that the diaryl sulfide is substituted. 7. Metoda prema zahtjevu 5, naznačena time što je diaril sulfid spoj koji je označen općom formulom (I): [image] (I) gdje su A i B, neovisno, arilne skiupine odabrane iz skupa kojega sačinjavaju 5- i 6-člani aormatski prsteni, uključujući i ne ograničavajući se samo na njih – fenil, tienil, furil, pirimidinil, imidazolil, pirazolil, piridil, pirazinil, pirolil i piridazinil; R1, R2 i R3 su neovisno odabrani iz skupa kojega sačinjavaju vodik, -Ra, gdje je Ra vodik ili alkilna skupina koja sadrži od jedan od šest lančastih ili razgranatih ugljikovih atoma (C1-6 alkil), -halogen, gdje je halogen Cl, F, Br ili I, -NRbRc, gdje su Rb i Rc neovisno H, C1-6 alkil ili –CH2-aril, -NO2, -C(=O)Ra, -CN, -perfluorRa, kao trifluormetil, -N-C(=O)Ra, -(CH2)n-NRbRc, gdje je n cijeli broj od 1 do 6, 5- ili 6-člani heterociklički prsten, bilo alifatski ili aromatski, koji sadrži jedan ili više O, N ili S, proizvoljno supstituiranih, kao što su morfolino, i –S-aril, gdje je aril 5- ili 6-člani aromatski prsten, proizvoljno supstituiran; a R4, R5 i R6, neovisno, odabrani su iz skupa kojega sačinjavaju vodik, -Ra, -O-Ra, -halo, -NRbRc, -NO2, -C(=O)Ra, -CN, -perfluorRa, -N-C(=O)Ra, -(CH2)n-NRbRc, i - 5- ili 6-člani heterociklički prsten, alifatski ili aromatski, koji sadrži jedan ili više O, N ili S, proizvoljno supstituiran, -S-aril, ili gdje R4 i R5 su uzeti zajedno da sačinjavaju 5- ili 6-člani aromatski prsten, koji proizvoljno sadrži jedan ili više O, N ili S u prstenu, proizvoljno supstituiran.7. The method according to claim 5, characterized in that the diaryl sulfide is a compound denoted by the general formula (I): [image] (AND) where A and B are, independently, aryl groups selected from the group consisting of 5- and 6-membered aromatic rings, including but not limited to - phenyl, thienyl, furyl, pyrimidinyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrrolyl and pyridazinyl; R1, R2 and R3 are independently selected from the set they comprise hydrogen, -Ra, where Ra is hydrogen or an alkyl group containing one of six chain or branched carbon atoms (C1-6 alkyl), -halogen, where halogen is Cl, F, Br or I, -NRbRc, where Rb and Rc are independently H, C1-6 alkyl or –CH2-aryl, -NO2, -C(=O)Ra, -CN, -perfluoroRa, as trifluoromethyl, -N-C(=O)Ra, -(CH2)n-NRbRc, where n is an integer from 1 to 6, A 5- or 6-membered heterocyclic ring, either aliphatic or aromatic, containing one or more O, N or S, optionally substituted, such as morpholino, and -S-aryl, where aryl is a 5- or 6-membered aromatic ring , arbitrarily substituted; and R 4 , R 5 and R 6 , independently, are selected from the set they comprise hydrogen, -Ra, -O-Ra, -halo, -NRbRc, -NO2, -C(=O)Ra, -CN, -perfluoroRa, -N-C(=O)Ra, -(CH2)n-NRbRc, i - 5- or 6-membered heterocyclic ring, aliphatic or aromatic, containing one or more O, N or S, arbitrarily substituted, -S-aryl, or where R 4 and R 5 are taken together to form a 5- or 6-membered aromatic ring, optionally containing one or more O, N or S in the ring, optionally substituted. 8. Metoda prema zahtjevu 7, naznačena time što je diaril sulfid odabran iz skupa kojega sačinjavaju: 3-Klor-4-(2-klorfenilsulfanil)-fenilamin hidroklorid 4-Nitro-2-klorfenil-(2,3-diklorfenil)-sulfid 3-Klor-4-(2-naftilsulfanil)-fenilamin hidroklorid 3-Klor-4-(2,3-diklorfenilsulfanil)-fenilamin hidroklorid 3-Klor-4-(2,4,5-triklorfenilsulfanil)-fenilamin hidroklorid 3-Klor-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 4-(Benzotiazol-2-ilsulfanil)-3-klor-fenilamin 3-Klor-4-(1-klor-naftalen-2-ilsulfanil)-fenilamin 3-Metoksi-4-(2,3-diklorfenilsulfanil)-fenilamin 5-Amino-2-(2,3-diklorfenilsulfanil)-acetofenon hidroklorid 4-(2,3-diklorfenilsulfanil)-fenilamin 3-Klor-4-(1-naftilsulfanil)-fenilamin hidroklorid 3-Metil-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 1-Acetamido-3-klor-4-(2,3-diklorfenilsulfanil)-benzen 4-Metilamino-2,2',4'-triklordifenilsulfid 3-Brom-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 3-Hidroksi-4-(2,3-diklorfenilsulfanil)-fenilamin hidroklorid 6-Klor-5-(2,4-diklorfenilsulfanil)-1 H-benzimidazole 4-Amino-2-klorfenil-(2',4'-dimetilfenil)-sulfid hidroklorid 2,5-Diklor-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 4-Amino-2-klorfenil-(2'-metil-4'-klorfenil)-sulfid 4-Amino-2-klorfenil-(2',4'-difluorfenil)-sulfid hidroklorid 4-Amino-2-klorfenil-(2',4',6'-triklorfenil)-sulfid 4-Amino-2-klorfenil-(2'-amino-4'-klorfenil)-sulfid 4-Amino-2-klorfenil-(2'-klor-4'-nitrofenil)-sulfid 4-Amino-2-klorfenil-(2'-nitro-4'-klorfenil)-sulfid 4-Amino-2-klorfenil-(3',4'-diklorfenil)-sulfid 4-Amino-2-klorfenil-2-(3-klor-5-trifluormetilpiridil)-sulfid Bis-(4,4'-diamino-2,2'-diklorfenil)-sulfid 4-Amino-2-klorfenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-klorfenil-(4'-acetamido-2'-klorfenil)-sulfid 4-Amino-2-klorfenil-6-(5-nitrokinolino)-sulfid 4-Amino-2-klorfenil-(4'-dimetilamino-2'-klorfenil)-sulfid 2-Klor-4-amino-5-metilaminofenil-(2',4'-diklorfenil)-sulfid 2-Klor-4-amino-5-N-morfolinofenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-trifluormetilfenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-klorfenil-2-(5-nitro-3-brom)-piridin sulfid 4-Aminometil-2-klorfenil-(2', 4'-diklorfenil)-sulfid 4,5-Diklor-2-(2,4-diklorfenilsulfanil)-fenilamin 3,5-Diklor-4-(2,4-diklorfenilsulfanil)-fenilamin 2,3-Diklor-4-(2,4-diklorfenilsulfanil)-fenilamin 4-Amino-2-fluorfenil-(2',4'-diklorfenil)-sulfid 5-Amino-3-klorfenil-(2',4'-diklorfenil)-sulfid 3-Klor-4-(1-klor-naftalen-2-ilsulfanil)-fenilamin 1-(3-Nitro-4-fenilsulfanil)-fenil)-etanon 1-(3-Nitro-4-fenilsulfanil)-fenil)-etanon oksim 5-Trifluormetil-2-fenilsulfanil-benzonitril 1-(3,5-diklorfenil)-3-fenilsulfanil-pirolidin-2,5-dion Bis-2,4,6-Trinitrofenil-sulfid 2-Metil-1-(2-o-tolilsulfanil-fenil)-1H-pirol 2-[2-(4-Klor-2-nitro-fenilsulfanil)-fenilamino-3H-izobenzofuran-1on 4-(Benzotiazol-2-ilsulfanil)-3-klor-fenilamin 2-Nitro-4-klorfenil-(2’-aminofenil)-sulfid 6-Amino-2-klorfenil-(4’-metilfenil)-sulfid 4-Nitrofenil-(2’-klorfenil)-sulfid 2,4-Dinitrofenil-(4’-klorfenil)-sulfid 4-Aminofenil-(2’-klorfenil)-sulfid 2,4-Diaminofenil-(4’-izopropilfenil)-sulfid 4-Nitro-2-klorfenil-(2’,3’-diklorfenil)-sulfid 4-Amino-2-klorfenil-2-(5-nitro-3-brom)-piridin-sulfid8. The method according to claim 7, characterized in that the diaryl sulfide is selected from the group consisting of: 3-Chloro-4-(2-chlorophenylsulfanyl)-phenylamine hydrochloride 4-Nitro-2-chlorophenyl-(2,3-dichlorophenyl)-sulfide 3-Chloro-4-(2-naphthylsulfanyl)-phenylamine hydrochloride 3-Chloro-4-(2,3-dichlorophenylsulfanyl)-phenylamine hydrochloride 3-Chloro-4-(2,4,5-trichlorophenylsulfanyl)-phenylamine hydrochloride 3-Chloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride 4-(Benzothiazol-2-ylsulfanyl)-3-chloro-phenylamine 3-Chloro-4-(1-chloro-naphthalen-2-ylsulfanyl)-phenylamine 3-Methoxy-4-(2,3-dichlorophenylsulfanyl)-phenylamine 5-Amino-2-(2,3-dichlorophenylsulfanyl)-acetophenone hydrochloride 4-(2,3-dichlorophenylsulfanyl)-phenylamine 3-Chloro-4-(1-naphthylsulfanyl)-phenylamine hydrochloride 3-Methyl-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride 1-Acetamido-3-chloro-4-(2,3-dichlorophenylsulfanyl)-benzene 4-Methylamino-2,2',4'-trichlorodiphenylsulfide 3-Bromo-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride 3-Hydroxy-4-(2,3-dichlorophenylsulfanyl)-phenylamine hydrochloride 6-Chloro-5-(2,4-dichlorophenylsulfanyl)-1H-benzimidazoles 4-Amino-2-chlorophenyl-(2',4'-dimethylphenyl)-sulfide hydrochloride 2,5-Dichloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride 4-Amino-2-chlorophenyl-(2'-methyl-4'-chlorophenyl)-sulfide 4-Amino-2-chlorophenyl-(2',4'-difluorophenyl)-sulfide hydrochloride 4-Amino-2-chlorophenyl-(2',4',6'-trichlorophenyl)sulfide 4-Amino-2-chlorophenyl-(2'-amino-4'-chlorophenyl)-sulfide 4-Amino-2-chlorophenyl-(2'-chloro-4'-nitrophenyl)-sulfide 4-Amino-2-chlorophenyl-(2'-nitro-4'-chlorophenyl)-sulfide 4-Amino-2-chlorophenyl-(3',4'-dichlorophenyl)-sulfide 4-Amino-2-chlorophenyl-2-(3-chloro-5-trifluoromethylpyridyl)-sulfide Bis-(4,4'-diamino-2,2'-dichlorophenyl)-sulfide 4-Amino-2-chlorophenyl-(2',4'-dichlorophenyl)-sulfide 4-Amino-2-chlorophenyl-(4'-acetamido-2'-chlorophenyl)-sulfide 4-Amino-2-chlorophenyl-6-(5-nitroquinolino)-sulfide 4-Amino-2-chlorophenyl-(4'-dimethylamino-2'-chlorophenyl)-sulfide 2-Chloro-4-amino-5-methylaminophenyl-(2',4'-dichlorophenyl)-sulfide 2-Chloro-4-amino-5-N-morpholinophenyl-(2',4'-dichlorophenyl)-sulfide 4-Amino-2-trifluoromethylphenyl-(2',4'-dichlorophenyl)-sulfide 4-Amino-2-chlorophenyl-2-(5-nitro-3-bromo)-pyridine sulfide 4-Aminomethyl-2-chlorophenyl-(2', 4'-dichlorophenyl)-sulfide 4,5-Dichloro-2-(2,4-dichlorophenylsulfanyl)-phenylamine 3,5-Dichloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine 2,3-Dichloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine 4-Amino-2-fluorophenyl-(2',4'-dichlorophenyl)-sulfide 5-Amino-3-chlorophenyl-(2',4'-dichlorophenyl)-sulfide 3-Chloro-4-(1-chloro-naphthalen-2-ylsulfanyl)-phenylamine 1-(3-Nitro-4-phenylsulfanyl)-phenyl)-ethanone 1-(3-Nitro-4-phenylsulfanyl)-phenyl)-ethanone oxime 5-Trifluoromethyl-2-phenylsulfanyl-benzonitrile 1-(3,5-dichlorophenyl)-3-phenylsulfanyl-pyrrolidine-2,5-dione Bis-2,4,6-Trinitrophenyl-sulphide 2-Methyl-1-(2-o-tolylsulfanyl-phenyl)-1H-pyrrole 2-[2-(4-Chloro-2-nitro-phenylsulfanyl)-phenylamino-3H-isobenzofuran-1one 4-(Benzothiazol-2-ylsulfanyl)-3-chloro-phenylamine 2-Nitro-4-chlorophenyl-(2'-aminophenyl)-sulfide 6-Amino-2-chlorophenyl-(4'-methylphenyl)-sulfide 4-Nitrophenyl-(2'-chlorophenyl)-sulfide 2,4-Dinitrophenyl-(4'-chlorophenyl)-sulfide 4-Aminophenyl-(2'-chlorophenyl)-sulfide 2,4-Diaminophenyl-(4'-isopropylphenyl)-sulfide 4-Nitro-2-chlorophenyl-(2',3'-dichlorophenyl)-sulfide 4-Amino-2-chlorophenyl-2-(5-nitro-3-bromo)-pyridine-sulfide 9. Metode za tretiranje upalne bolesti koja nastaje zbog LFA-1 vezanja za njegov prirodni ligand koji kompetira s ICAM-1 ili ICAM-3 za vezanje na LFA-1, naznačena time što uključuje primjenu sisavcu kojemu je to potrebno spoja koji kompetira s 3-klor-4-(1-klor-naftalen-2-ilsulfanil)-fenilaminom za vezanje na LFA-1 u količini koja je dovoljna da inhibira vezanje prirodnog liganda za LFA-1.9. Methods for treating an inflammatory disease caused by LFA-1 binding to its natural ligand that competes with ICAM-1 or ICAM-3 for binding to LFA-1, comprising administering to a mammal in need thereof a compound that competes with 3 -chloro-4-(1-chloro-naphthalen-2-ylsulfanyl)-phenylamine for binding to LFA-1 in an amount sufficient to inhibit binding of the natural ligand to LFA-1. 10. Metoda prema zahtjevu 9, naznačena time što je spoj diaril sulfid.10. The method according to claim 9, characterized in that the compound is a diaryl sulfide. 11. Metoda prema zahtjevu 10, naznačena time što je diaril sulfid supstituiran.11. The method according to claim 10, characterized in that the diaryl sulfide is substituted. 12. Metoda prema zahtjevu 10, naznačena time što je diaril sulfid spoj označen općom formulom (I): [image] (I) gdje su A i B, neovisno, arilne skiupine odabrane iz skupa kojega sačinjavaju 5- i 6-člani aormatski prsteni, uključujući i ne ograničavajući se samo na njih – fenil, tienil, furil, pirimidinil, imidazolil, pirazolil, piridil, pirazinil, pirolil i piridazinil; R1, R2 i R3 su neovisno odabrani iz skupa kojega sačinjavaju vodik, -Ra, gdje je Ra vodik ili alkilna skupina koja sadrži od jedan od šest lančastih ili razgranatih ugljikovih atoma (C1-6 alkil), -halogen, gdje je halogen Cl, F, Br ili I, -NRbRc, gdje su Rb i Rc neovisno H, C1-6 alkil ili –CH2-aril, -NO2, -C(=O)Ra, -CN, -perfluorRa, kao trifluormetil, -N-C(=O)Ra, -(CH2)n-NRbRc, gdje je n cijeli broj od 1 do 6, 5- ili 6-člani heterociklički prsten, bilo alifatski ili aromatski, koji sadrži jedan ili više O, N ili S, proizvoljno supstituiranih, kao što su morfolino, i –S-aril, gdje je aril 5- ili 6-člani aromatski prsten, proizvoljno supstituiran; a R4, R5 i R6, neovisno, odabrani su iz skupa kojega sačinjavaju vodik, -Ra, -O-Ra, -halo, -NRbRc, -NO2, -C(=O)Ra, -CN, -perfluorRa, -N-C(=O)Ra, -(CH2)n-NRbRc, i - 5- ili 6-člani heterociklički prsten, alifatski ili aromatski, koji sadrži jedan ili više O, N ili S, proizvoljno supstituiran, -S-aril, ili gdje R4 i R5 su uzeti zajedno da sačinjavaju 5- ili 6-člani aromatski prsten, koji proizvoljno sadrži jedan ili više O, N ili S u prstenu, proizvoljno supstituiran.12. The method according to claim 10, characterized in that the diaryl sulfide is a compound denoted by the general formula (I): [image] (AND) where A and B are, independently, aryl groups selected from the group consisting of 5- and 6-membered aromatic rings, including but not limited to - phenyl, thienyl, furyl, pyrimidinyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrrolyl and pyridazinyl; R1, R2 and R3 are independently selected from the set they comprise hydrogen, -Ra, where Ra is hydrogen or an alkyl group containing one of six chain or branched carbon atoms (C1-6 alkyl), -halogen, where halogen is Cl, F, Br or I, -NRbRc, where Rb and Rc are independently H, C1-6 alkyl or –CH2-aryl, -NO2, -C(=O)Ra, -CN, -perfluoroRa, as trifluoromethyl, -N-C(=O)Ra, -(CH2)n-NRbRc, where n is an integer from 1 to 6, A 5- or 6-membered heterocyclic ring, either aliphatic or aromatic, containing one or more O, N or S, optionally substituted, such as morpholino, and -S-aryl, where aryl is a 5- or 6-membered aromatic ring , arbitrarily substituted; and R 4 , R 5 and R 6 , independently, are selected from the set they comprise hydrogen, -Ra, -O-Ra, -halo, -NRbRc, -NO2, -C(=O)Ra, -CN, -perfluoroRa, -N-C(=O)Ra, -(CH2)n-NRbRc, i - 5- or 6-membered heterocyclic ring, aliphatic or aromatic, containing one or more O, N or S, arbitrarily substituted, -S-aryl, or where R 4 and R 5 are taken together to form a 5- or 6-membered aromatic ring, optionally containing one or more O, N or S in the ring, optionally substituted. 13. Metoda prema zahtjevu 12, naznačena time što je diaril sulfid odabran iz skupa kojega sačinjavaju: 3-Klor-4-(2-klorfenilsulfanil)-fenilamin hidroklorid 4-Nitro-2-klorfenil-(2,3-diklorfenil)-sulfid 3-Klor-4-(2-naftilsulfanil)-fenilamin hidroklorid 3-Klor-4-(2,3-diklorfenilsulfanil)-fenilamin hidroklorid 3-Klor-4-(2,4,5-triklorfenilsulfanil)-fenilamin hidroklorid 3-Klor-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 4-(Benzotiazol-2-ilsulfanil)-3-klor-fenilamin 3-Klor-4-(1-klor-naftalen-2-ilsulfanil)-fenilamin 3-Metoksi-4-(2,3-diklorfenilsulfanil)-fenilamin 5-Amino-2-(2,3-diklorfenilsulfanil)-acetofenon hidroklorid 4-(2,3-diklorfenilsulfanil)-fenilamin 3-Klor-4-(1-naftilsulfanil)-fenilamin hidroklorid 3-Metil-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 1-Acetamido-3-klor-4-(2,3-diklorfenilsulfanil)-benzen 4-Metilamino-2,2',4'-triklordifenilsulfid 3-Brom-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 3-Hidroksi-4-(2,3-diklorfenilsulfanil)-fenilamin hidroklorid 6-Klor-5-(2,4-diklorfenilsulfanil)-1 H-benzimidazole 4-Amino-2-klorfenil-(2',4'-dimetilfenil)-sulfid hidroklorid 2,5-Diklor-4-(2,4-diklorfenilsulfanil)-fenilamin hidroklorid 4-Amino-2-klorfenil-(2'-metil-4'-klorfenil)-sulfid 4-Amino-2-klorfenil-(2',4'-difluorfenil)-sulfid hidroklorid 4-Amino-2-klorfenil-(2',4',6'-triklorfenil)-sulfid 4-Amino-2-klorfenil-(2'-amino-4'-klorfenil)-sulfid 4-Amino-2-klorfenil-(2'-klor-4'-nitrofenil)-sulfid 4-Amino-2-klorfenil-(2'-nitro-4'-klorfenil)-sulfid 4-Amino-2-klorfenil-(3',4'-diklorfenil)-sulfid 4-Amino-2-klorfenil-2-(3-klor-5-trifluormetilpiridil)-sulfid Bis-(4,4'-diamino-2,2'-diklorfenil)-sulfid 4-Amino-2-klorfenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-klorfenil-(4'-acetamido-2'-klorfenil)-sulfid 4-Amino-2-klorfenil-6-(5-nitrokinolino)-sulfid 4-Amino-2-klorfenil-(4'-dimetilamino-2'-klorfenil)-sulfid 2-Klor-4-amino-5-metilaminofenil-(2',4'-diklorfenil)-sulfid 2-Klor-4-amino-5-N-morfolinofenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-trifluormetilfenil-(2',4'-diklorfenil)-sulfid 4-Amino-2-klorfenil-2-(5-nitro-3-brom)-piridin sulfid 4-Aminometil-2-klorfenil-(2', 4'-diklorfenil)-sulfid 4,5-Diklor-2-(2,4-diklorfenilsulfanil)-fenilamin 3,5-Diklor-4-(2,4-diklorfenilsulfanil)-fenilamin 2,3-Diklor-4-(2,4-diklorfenilsulfanil)-fenilamin 4-Amino-2-fluorfenil-(2',4'-diklorfenil)-sulfid 5-Amino-3-klorfenil-(2',4'-diklorfenil)-sulfid 3-Klor-4-(1-klor-naftalen-2-ilsulfanil)-fenilamin 1-(3-Nitro-4-fenilsulfanil)-fenil)-etanon 1-(3-Nitro-4-fenilsulfanil)-fenil)-etanon oksim 5-Trifluormetil-2-fenilsulfanil-benzonitril 1-(3,5-diklorfenil)-3-fenilsulfanil-pirolidin-2,5-dion Bis-2,4,6-Trinitrofenil-sulfid 2-Metil-1-(2-o-tolilsulfanil-fenil)-1H-pirol 2-[2-(4-Klor-2-nitro-fenilsulfanil)-fenilamino-3H-izobenzofuran-1on 4-(Benzotiazol-2-ilsulfanil)-3-klor-fenilamin 2-Nitro-4-klorfenil-(2’-aminofenil)-sulfid 6-Amino-2-klorfenil-(4’-metilfenil)-sulfid 4-Nitrofenil-(2’-klorfenil)-sulfid 2,4-Dinitrofenil-(4’-klorfenil)-sulfid 4-Aminofenil-(2’-klorfenil)-sulfid 2,4-Diaminofenil-(4’-izopropilfenil)-sulfid 4-Nitro-2-klorfenil-(2’,3’-diklorfenil)-sulfid 4-Amino-2-klorfenil-2-(5-nitro-3-brom)-piridin-sulfid13. The method according to claim 12, characterized in that the diaryl sulfide is selected from the group consisting of: 3-Chloro-4-(2-chlorophenylsulfanyl)-phenylamine hydrochloride 4-Nitro-2-chlorophenyl-(2,3-dichlorophenyl)-sulfide 3-Chloro-4-(2-naphthylsulfanyl)-phenylamine hydrochloride 3-Chloro-4-(2,3-dichlorophenylsulfanyl)-phenylamine hydrochloride 3-Chloro-4-(2,4,5-trichlorophenylsulfanyl)-phenylamine hydrochloride 3-Chloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride 4-(Benzothiazol-2-ylsulfanyl)-3-chloro-phenylamine 3-Chloro-4-(1-chloro-naphthalen-2-ylsulfanyl)-phenylamine 3-Methoxy-4-(2,3-dichlorophenylsulfanyl)-phenylamine 5-Amino-2-(2,3-dichlorophenylsulfanyl)-acetophenone hydrochloride 4-(2,3-dichlorophenylsulfanyl)-phenylamine 3-Chloro-4-(1-naphthylsulfanyl)-phenylamine hydrochloride 3-Methyl-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride 1-Acetamido-3-chloro-4-(2,3-dichlorophenylsulfanyl)-benzene 4-Methylamino-2,2',4'-trichlorodiphenylsulfide 3-Bromo-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride 3-Hydroxy-4-(2,3-dichlorophenylsulfanyl)-phenylamine hydrochloride 6-Chloro-5-(2,4-dichlorophenylsulfanyl)-1H-benzimidazoles 4-Amino-2-chlorophenyl-(2',4'-dimethylphenyl)-sulfide hydrochloride 2,5-Dichloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine hydrochloride 4-Amino-2-chlorophenyl-(2'-methyl-4'-chlorophenyl)-sulfide 4-Amino-2-chlorophenyl-(2',4'-difluorophenyl)-sulfide hydrochloride 4-Amino-2-chlorophenyl-(2',4',6'-trichlorophenyl)sulfide 4-Amino-2-chlorophenyl-(2'-amino-4'-chlorophenyl)-sulfide 4-Amino-2-chlorophenyl-(2'-chloro-4'-nitrophenyl)-sulfide 4-Amino-2-chlorophenyl-(2'-nitro-4'-chlorophenyl)-sulfide 4-Amino-2-chlorophenyl-(3',4'-dichlorophenyl)-sulfide 4-Amino-2-chlorophenyl-2-(3-chloro-5-trifluoromethylpyridyl)-sulfide Bis-(4,4'-diamino-2,2'-dichlorophenyl)-sulfide 4-Amino-2-chlorophenyl-(2',4'-dichlorophenyl)-sulfide 4-Amino-2-chlorophenyl-(4'-acetamido-2'-chlorophenyl)-sulfide 4-Amino-2-chlorophenyl-6-(5-nitroquinolino)-sulfide 4-Amino-2-chlorophenyl-(4'-dimethylamino-2'-chlorophenyl)-sulfide 2-Chloro-4-amino-5-methylaminophenyl-(2',4'-dichlorophenyl)-sulfide 2-Chloro-4-amino-5-N-morpholinophenyl-(2',4'-dichlorophenyl)-sulfide 4-Amino-2-trifluoromethylphenyl-(2',4'-dichlorophenyl)-sulfide 4-Amino-2-chlorophenyl-2-(5-nitro-3-bromo)-pyridine sulfide 4-Aminomethyl-2-chlorophenyl-(2', 4'-dichlorophenyl)-sulfide 4,5-Dichloro-2-(2,4-dichlorophenylsulfanyl)-phenylamine 3,5-Dichloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine 2,3-Dichloro-4-(2,4-dichlorophenylsulfanyl)-phenylamine 4-Amino-2-fluorophenyl-(2',4'-dichlorophenyl)-sulfide 5-Amino-3-chlorophenyl-(2',4'-dichlorophenyl)-sulfide 3-Chloro-4-(1-chloro-naphthalen-2-ylsulfanyl)-phenylamine 1-(3-Nitro-4-phenylsulfanyl)-phenyl)-ethanone 1-(3-Nitro-4-phenylsulfanyl)-phenyl)-ethanone oxime 5-Trifluoromethyl-2-phenylsulfanyl-benzonitrile 1-(3,5-dichlorophenyl)-3-phenylsulfanyl-pyrrolidine-2,5-dione Bis-2,4,6-Trinitrophenyl-sulphide 2-Methyl-1-(2-o-tolylsulfanyl-phenyl)-1H-pyrrole 2-[2-(4-Chloro-2-nitro-phenylsulfanyl)-phenylamino-3H-isobenzofuran-1one 4-(Benzothiazol-2-ylsulfanyl)-3-chloro-phenylamine 2-Nitro-4-chlorophenyl-(2'-aminophenyl)-sulfide 6-Amino-2-chlorophenyl-(4'-methylphenyl)-sulfide 4-Nitrophenyl-(2'-chlorophenyl)-sulfide 2,4-Dinitrophenyl-(4'-chlorophenyl)-sulfide 4-Aminophenyl-(2'-chlorophenyl)-sulfide 2,4-Diaminophenyl-(4'-isopropylphenyl)-sulfide 4-Nitro-2-chlorophenyl-(2',3'-dichlorophenyl)-sulfide 4-Amino-2-chlorophenyl-2-(5-nitro-3-bromo)-pyridine-sulfide 14. Metoda za inhibiranje vezanja LFA-1 za svoj prirodni ligand koja kompetira sa ICAM-1 ili ICAM-3 za vezanje sa LFA-1, naznačena time što obuhvaća dodir stanice koja vrši ekspresiju LFA-1 na svojoj površini saspojem koji kompetira s 3-klor-4-(1-klor-naftalen-2-ilsulfanil)-fenilaminom za vezanje s LFA-1 u količini koja je dovoljna za inhibiranje vezanja LFA-1 s prirodnim ligandom.14. A method for inhibiting the binding of LFA-1 to its natural ligand that competes with ICAM-1 or ICAM-3 for binding to LFA-1, characterized in that it comprises contacting a cell that expresses LFA-1 on its surface with a compound that competes with 3 -chloro-4-(1-chloro-naphthalen-2-ylsulfanyl)-phenylamine for binding to LFA-1 in an amount sufficient to inhibit binding of LFA-1 to the natural ligand. 15. Metoda prema zahtjevu 9 ili 14, naznačena time što je ligand ICAM-1 ili ICAM-3.15. The method according to claim 9 or 14, characterized in that the ligand is ICAM-1 or ICAM-3. 16. Metoda za identificiranje negativnog regulatora LFA-1 vezanja sa svojim prirodnim ligandom koji kompetira s ICAM-1 ili ICAM-3 za vezanje s LFA-1, naznačena time što uključuje sljedeće stupnjeve: a) kontaktiranje LFA-1 s aktivatorom LFA-1 vezanja; b) mjerenje LFA-1 vezanja s prirodnim ligandom u prisutnosti i odsutnosti spoja koji se ispituje; i c) identificiranje spoja koji se ispituje kao negativni regulator kada se smanjeno vezanje LFA-1 za ligand uočava u prisutnosti spoja koji se ispituje.16. A method for identifying a negative regulator of LFA-1 binding with its natural ligand that competes with ICAM-1 or ICAM-3 for binding with LFA-1, characterized in that it includes the following steps: a) contacting LFA-1 with the LFA-1 binding activator; b) measuring LFA-1 binding to the native ligand in the presence and absence of the test compound; and c) identifying the test compound as a negative regulator when reduced binding of LFA-1 to the ligand is observed in the presence of the test compound. 17. Metoda prema zahtjevu 16, naznačena time što je aktivator kristalno ljubičasto (crystal violet).17. The method according to claim 16, characterized in that the activator is crystal violet. 18. Spoj koji je supstituirani diarilni sulfid ili njegova farmaceutski prihvatljiva sol ili prolijek, naznačen time što se veže za domenu I LFA-1 kao preferentnu u odnosu na ICAM.18. A compound which is a substituted diaryl sulfide or a pharmaceutically acceptable salt or prodrug thereof, characterized in that it binds to domain I of LFA-1 preferentially over ICAM. 19. Spoj koji je supstituirani diarilni sulfid formule (I): [image] (I) ili njegova farmaceutski prihvatljiva sol ili prolijek, naznačeni time što: A i B su, neovisno, arilne skupine odabrane iz skupa kojega sačinjavaju 5- i 6-člani aromatski prsteni, uključujući i ne ograničavajući se samo na njih – fenil, tienil, furil, pirimidinil, imidazolil, pirazolil, piridil, pirazinil, pirolil i piridazinil; R1, R2 i R3 su neovisno odabrani iz skupa kojega sačinjavaju vodik, -Ra, gdje je Ra vodik ili alkilna skupina koja sadrži od jedan od šest lančastih ili razgranatih ugljikovih atoma (C1-6 alkil), -halogen, gdje je halogen Cl, F, Br ili I, -NRbRc, gdje su Rb i Rc neovisno H, C1-6 alkil ili –CH2-aril, -NO2, -C(=O)Ra, -CN, -perfluorRa, kao trifluormetil, -N-C(=O)Ra, -(CH2)n-NRbRc, gdje je n cijeli broj od 1 do 6, 5- ili 6-člani heterociklički prsten, bilo alifatski ili aromatski, koji sadrži jedan ili više O, N ili S, proizvoljno supstituiranih, kao što su morfolino, i –S-aril, gdje je aril 5- ili 6-člani aromatski prsten, proizvoljno supstituiran; a R4, R5 i R6, neovisno, odabrani su iz skupa kojega sačinjavaju vodik, -Ra, -O-Ra, -halo, -NRbRc, -NO2, -C(=O)Ra, -CN, -perfluorRa, -N-C(=O)Ra, -(CH2)n-NRbRc, i 5- ili 6-člani heterociklički prsten, alifatski ili aromatski, koji sadrži jedan ili više O, N ili S, proizvoljno supstituiran, -S-aril, ili gdje R4 i R5 su uzeti zajedno da sačinjavaju 5- ili 6-člani aromatski prsten, koji proizvoljno sadrži jedan ili više O, N ili S u prstenu, proizvoljno supstituiran.19. A compound which is a substituted diaryl sulfide of formula (I): [image] (AND) or a pharmaceutically acceptable salt or prodrug thereof, characterized in that: A and B are, independently, aryl groups selected from the group consisting of 5- and 6-membered aromatic rings, including but not limited to - phenyl, thienyl, furyl, pyrimidinyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrrolyl and pyridazinyl; R1, R2 and R3 are independently selected from the set they comprise hydrogen, -Ra, where Ra is hydrogen or an alkyl group containing one of six chain or branched carbon atoms (C1-6 alkyl), -halogen, where halogen is Cl, F, Br or I, -NRbRc, where Rb and Rc are independently H, C1-6 alkyl or –CH2-aryl, -NO2, -C(=O)Ra, -CN, -perfluoroRa, as trifluoromethyl, -N-C(=O)Ra, -(CH2)n-NRbRc, where n is an integer from 1 to 6, A 5- or 6-membered heterocyclic ring, either aliphatic or aromatic, containing one or more O, N or S, optionally substituted, such as morpholino, and -S-aryl, where aryl is a 5- or 6-membered aromatic ring , arbitrarily substituted; and R 4 , R 5 and R 6 , independently, are selected from the set they comprise hydrogen, -Ra, -O-Ra, -halo, -NRbRc, -NO2, -C(=O)Ra, -CN, -perfluoroRa, -N-C(=O)Ra, -(CH2)n-NRbRc, i A 5- or 6-membered heterocyclic ring, aliphatic or aromatic, containing one or more O, N or S, optionally substituted, -S-aryl, or where R4 and R5 are taken together to form a 5- or 6-membered aromatic ring , which optionally contains one or more O, N or S in the ring, optionally substituted. 20. Metoda za tretiranje kliničkog stanja u sisavca, naznačena time što uključuje primjenu sisavcu terapeutski učinkovite količine spoja kao što je navedeno u zahtjevu 18 ili 19.20. A method for treating a clinical condition in a mammal, characterized in that it includes administering to the mammal a therapeutically effective amount of the compound as set forth in claim 18 or 19.
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