NO169964B

NO169964B - ANALOGY PROCEDURE FOR THE PREPARATION OF THERAPEUTICALLY ACTIVE ACYL-SUBSTITUTED N-BENZOPYRANAL LACTAMES

Info

Publication number: NO169964B
Application number: NO880474A
Authority: NO
Inventors: Heinrich Christian Englert; Hans-Jochen Lang; Dieter Mania; Bernward Schoelkens; Erik Klaus
Original assignee: Hoechst Ag
Priority date: 1987-02-04
Filing date: 1988-02-03
Publication date: 1992-05-18
Also published as: FI880468A0; PT86691A; KR880009957A; DK55188D0; FI92064B; DE3881714D1; IE60932B1; NZ223384A; IE880295L; FI880468A; NO880474L; DK55188A; EP0277611A3; ES2058148T3; IL85289A; PT86691B; AU1122788A; EP0277611B1; NO880474D0; HU207729B

Description

Foreliggende oppfinnelse vedrører en analogifremgangsmåte for fremstilling av N-benzopyranyllaktamer med formel I, The present invention relates to an analogous process for the production of N-benzopyranyl lactams of formula I,

hvori in which

R<1> betyr CN, S02-( C1- Cb )-alkyl eller S02-Ph, R<1> means CN, SO2-(C1-Cb)-alkyl or SO2-Ph,

R<2> betyr H, ( C^-Cg )-alkoksy, R<2> means H, (C 1 -C 8 )-Alkoxy,

R<3> og R<4> er like og betyr alkyl med 1-4 C-atomer, R<3> and R<4> are the same and mean alkyl with 1-4 C atoms,

X betyr en kjede (CH2)m, som er substituert med en (C1-C2)-alkylgruppe og m betyr 3 eller 4, idet OH-gruppen i 3-stillingen og laktamgruppen i 4-stillingen alltid har transkonfigurasj on. X means a chain (CH2)m, which is substituted with a (C1-C2)-alkyl group and m means 3 or 4, the OH group in the 3-position and the lactam group in the 4-position always having a trans configuration.

C-atomene 3 og 4 av 3,4-dihydro-2H-benzo[b]pyransystemet (nedenfor også betegnet som "kromansysternet") av formel I er substituert asymmetrisk. Oppfinnelsen vedrører bare slike forbindelser som ved disse sentere har motsatte konfigura-sjoner. Dette betyr at laktamringen som substituent ved C-4 og OH-gruppen ved C-3 alltid er orientert "trans" til hverandre. Den ovennevnte definisjon av X betyr, at dessuten laktamringen minst inneholder et, maksimalt imidlertid m (med m i den innledningsvis nevnte definisjon) chirale C-atomer. Oppfinnelsen vedrører derved såvel forbindelse med R-, som også S-konfigurerte sentere. Det samme gjelder for det tilfelle at R<1>, R<2> og R<3> eller R<4> inneholder asymmetrisentere eller selv som substituenter frembringer et asymmetrisenter. Forbindelsene kan da foreligge som optiske isomere, som diastereoisomere, som rasemater eller som blandinger av disse. The carbon atoms 3 and 4 of the 3,4-dihydro-2H-benzo[b]pyran system (also referred to below as the "chroman ester") of formula I are substituted asymmetrically. The invention only relates to such compounds which at these centers have opposite configurations. This means that the lactam ring as a substituent at C-4 and the OH group at C-3 are always oriented "trans" to each other. The above definition of X means that, in addition, the lactam ring contains at least one, however at most m (with m in the initially mentioned definition) chiral C atoms. The invention thereby relates to connection with both R- and S-configured centres. The same applies to the case that R<1>, R<2> and R<3> or R<4> contain centers of asymmetry or even as substituents produce an center of asymmetry. The compounds can then exist as optical isomers, as diastereoisomers, as racemates or as mixtures of these.

I J. Med. Chem. 1986, 29, 2194-2201 omtales forbindelser som står nærmest forbindelsene fremstilt ifølge oppfinnelsen. De er der sammenfattet under følgende generelle formler: In J. Med. Chem. 1986, 29, 2194-2201 mentions compounds which are closest to the compounds produced according to the invention. They are there summarized under the following general formulas:

En stor del av disse forbindelser er også omtalt i forskjellige patentsøknader, og nevner herved: EP 107 423, EP 120 417, EP 076 075 og EP 120 428. A large part of these compounds are also mentioned in various patent applications, and hereby mention: EP 107 423, EP 120 417, EP 076 075 and EP 120 428.

Av disse forbindelser er det kjent, at de formår å senke et syklig forhøyet blodtrykk, idet de relakserer den glatte karmuskulatur resp. kan beskytte overfor pressorisk irrita-sjon eller gjøre ufølsomme. Of these compounds, it is known that they manage to lower a morbidly elevated blood pressure, as they relax the vascular smooth muscle resp. can protect against pressure irritation or make insensitive.

Med forbindelsene fremstilt ifølge oppfinnelsen med formel I ble det nå funnet en ny stoffklasse med blodtrykksenkende egenskaper, som adskiller seg fra det kjente, frem for alt ved, at de i laktamringen som substituent ved C-4 av kromansystemet har ekstra substituenter med ovennevnte definisjon. Denne substitusjon fører sammenlignet med de allerede kjente usubstituerte forbindelser, til en betraktlig økning av den antihypertensive tilleggsvirkning. Dessuten ble det iakttatt at det i noen tilfeller følges en slik virkningsøkning og en nedsettelse av den akutte toksisitet, hvorav det tilsammen lar seg avlede en forbedring av det terapeutiske bredde. Dette er nettopp av fremragende betydning ved en langtidsterapi som hypertonibehandlingen, hvor et medikament må inntas under tiden hele livet. With the compounds produced according to the invention with formula I, a new class of substances with blood pressure-lowering properties was now found, which differs from the known, above all in that in the lactam ring as a substituent at C-4 of the chroman system, they have additional substituents with the above definition. Compared to the already known unsubstituted compounds, this substitution leads to a considerable increase in the additional antihypertensive effect. In addition, it was observed that in some cases such an increase in effectiveness and a reduction in acute toxicity are followed, from which together it is possible to derive an improvement in the therapeutic breadth. This is precisely of outstanding importance in a long-term therapy such as the treatment of hypertension, where a drug must be taken during the whole life.

De steriske forutsetninger, som fører til en slik fordelaktig profil krever nærmere forklaring. Dette kunne i ovennevnte 1itteratursitat vise, at for en optimal antihypertensiv virkning, må substituentene ved C-3 og C-4 være anordnet til hverandre i trans-stilling. En cis-anordning førte derimot til en tydelig svekning av virkningen. Således trans-konfigurerte forbindelser opptrer nå i tillegg dessuten i form av antipoder. Ved hjelp av et eksempel kunne det vises at (- )-antipoden er meget sterkere virksom enn ( + )-antipoden. Til (-)-antipoden ble det derved tilordnet 4R,3S-konfigurasjon (sammenlign EP 0 120 428). The steric assumptions that lead to such an advantageous profile require further explanation. This could show in the above-mentioned literature quote that for an optimal antihypertensive effect, the substituents at C-3 and C-4 must be arranged to each other in the trans position. A cis device, on the other hand, led to a clear weakening of the effect. Compounds thus trans-configured now also appear in the form of antipodes. Using an example, it could be shown that the (-) antipode is much more effective than the ( + ) antipode. The 4R,3S configuration was thereby assigned to the (-)-antipode (compare EP 0 120 428).

Ved innføring av substituenter i laktamringen i hht. formel I frembringes nå ytterligere asymmetrisentere. Forbindelsene fremstilt ifølge oppfinnelsen kan da ikke bare opptre som antipoder, men man iakttar i tillegg opptreden av diastereoisomere, som ved identisk konstitusjonsformer absolutt er adskillbare fra hverandre ved hjelp av vanlig spektro-skopiske eller .kromatografiske metoder. Konfigurasjonen av et nytt asymmetrisentrum i laktamringen kan nå med hensyn til f.eks. konfigurasjonen av C-4 i kromansystemet være lik eller motsatt. Er den lik, så er den med hensyn til konfigurasjonen ved C-3 motsatt, er den motsatt så er den med hensyn til C-3 lik. Disse to relative anordninger av en bestemt konfigurasjon i laktamringen, med hensyn til trans-konfigurasjonen av kromansystemet manifiserer seg i 2 diastereomere forbindelser med formel I. Overraskende iakttas nå i mange tilfeller de ovennevnte fordelaktige antihypertensive egenskaper av forbindelsen I bare for en av de to diastereomere, mens den andre er meget virksom i forhold til den tilsvarende usubstituerte forbindelse. Foretrukket er dette slike forbindelser, som har en alkylsubstituent ved C-atomet av laktamringen som er naboplassert til laktam-nitrogenet og hvis relative konfigurasjon er identisk med konfigurasjonen av C-4 atomet i kromansystemet. When introducing substituents in the lactam ring in accordance with formula I, additional centers of asymmetry are now produced. The compounds produced according to the invention can then not only act as antipodes, but one also observes the appearance of diastereoisomers, which, with identical constitutional forms, are absolutely separable from each other by means of usual spectroscopic or chromatographic methods. The configuration of a new center of asymmetry in the lactam ring can reach with regard to e.g. the configuration of C-4 in the chromane system be equal or opposite. If it is equal, then with respect to the configuration at C-3 it is opposite, if it is opposite then it is equal with respect to C-3. These two relative arrangements of a particular configuration in the lactam ring, with respect to the trans-configuration of the chromane system, manifest themselves in 2 diastereomeric compounds of formula I. Surprisingly, in many cases, the above-mentioned beneficial antihypertensive properties of compound I are now observed only for one of the two diastereomers , while the other is very active compared to the corresponding unsubstituted compound. These are preferred compounds which have an alkyl substituent at the C atom of the lactam ring which is adjacent to the lactam nitrogen and whose relative configuration is identical to the configuration of the C-4 atom in the chromane system.

Innføring av substituenter i laktamringen i hht. formel I fører således ikke som man kunne ha ventet, i alle tilfelle til en brukbare antihypertensiv virkning, men det er å overholde en helt spesiell rommelig orientering av disse substituenter i forhold til de allerede tilstedeværende asymmetrisentere. Noen av forbindelsene med formel I adskiller seg videre fra de allerede kjente forbindelser også ved, at de i tillegg som substituenter i benzodelen har en arylsulfonyl- eller en arylsulfoksysubstituent med aryl i innledningsvis nevnte betydning. Det var tidligere ikke kjent, at også en slik substitusjon kan føre til denne antihypertensive virkning. Introduction of substituents in the lactam ring according to formula I thus does not, as one might have expected, in any case lead to a usable antihypertensive effect, but it is to observe a very special spatial orientation of these substituents in relation to the already present asymmetry centers. Some of the compounds of formula I also differ from the already known compounds in that they also have an arylsulfonyl or an arylsulfoxy substituent with aryl in the meaning mentioned at the outset as substituents in the benzo part. It was previously not known that such a substitution can also lead to this antihypertensive effect.

Videre viser noen av forbindelsene fremstilt ifølge oppfinnelsen, spesielt slike diastereomere, som som nevnt ovenfor har mindre utpreget hjertekretsløpeffekter, en sterk relakserende effekt på uretere. Dermed er slike forbindelser I verdifulle terapeutika ved behandling av urinlederkoliker eller ved 1itotripsibehandling. I begge tilfeller letter en relakserende effekt på uretere en avgang av stener. Furthermore, some of the compounds produced according to the invention, especially such diastereomers, which, as mentioned above, have less pronounced cardiovascular effects, show a strong relaxing effect on the ureters. Thus, such compounds are valuable therapeutics in the treatment of ureteral colic or in isotripsy treatment. In both cases, a relaxing effect on the ureters facilitates the passage of stones.

Foreliggende oppfinnelse er følgelig kjennetegnet ved at The present invention is therefore characterized by the fact that

a) forbindelser med formel II a) compounds of formula II

hvori R<1>, R<2>, R<3> og R<4> har ovennevnte betydning, omsettes med laktamer med formel III in which R<1>, R<2>, R<3> and R<4> have the above meaning, are reacted with lactams of formula III

eller or

b) forbindelser med formel IV b) compounds of formula IV

hvori R<1>, R<2>, R<3> og R<4> har ovennevnte betydning, wherein R<1>, R<2>, R<3> and R<4> have the above meaning,

omsettes med laktamer med formel III. is reacted with lactams of formula III.

Fremstilles forbindelsene I etter metodene a) eller b), så foregår dette ved, at man omsetter forbindelsene IV eller II i et egnet oppløsningsmiddel, fortrinnsvis i dipolare aprotiske oppløsningsmidler, som dimetylsulfoksyd eller THF med laktamene III, fortrinnsvis under innvirkning av baser, som natriumhydrid, K-tert.butylat eller lignende, for laktam-N-alkyleringen kjent egnede baser. Reaksjonstemperaturen er dermed varierbar innen vide grenser, fortrinnsvis arbeides mellom 0' og værelsestemperatur. Ved anvendelse av rasemiske eller også optisk enhetlige laktamer III, fås derved minst to nye produkter med formel I. Disse produkter kan ved de vanlige metoder, som krystallisering eller kromatografering spaltes, i mange tilfeller har også en kombinasjon av begge metoder vist seg som gunstig. De respektive produkter kan deretter ved vanlige fysikalske undersøkelser, som f.eks. røntgenstrukturanalyse eller NMR-spektroskopi, tilordnes den resp. samlede konfigurasjon. Optisk enhetlige, altså enantiomerrene forbindelser I kan fås ved etterfølgende rasematspaltning. Det anvendes imidlertid allerede enantiomerrene laktamer I så får man de diastereomere forbindelser I likeledes i enantiomerren form og en rasematspaltning blir overflødig. If the compounds I are prepared according to methods a) or b), this takes place by reacting the compounds IV or II in a suitable solvent, preferably in dipolar aprotic solvents, such as dimethylsulfoxide or THF with the lactams III, preferably under the influence of bases, such as sodium hydride , K-tert.butylate or the like, known suitable bases for the lactam N-alkylation. The reaction temperature is thus variable within wide limits, preferably working between 0' and room temperature. By using racemic or also optically uniform lactams III, at least two new products of formula I are thereby obtained. These products can be cleaved by the usual methods, such as crystallization or chromatography, in many cases a combination of both methods has also proven beneficial. The respective products can then by normal physical examinations, such as e.g. X-ray structure analysis or NMR spectroscopy, it is assigned to the resp. overall configuration. Optically uniform, i.e. enantiomerically pure, compounds I can be obtained by subsequent racemate cleavage. However, if enantiomeric lactams I are already used, the diastereomeric compounds I are also obtained in enantiomeric form and a racemate cleavage becomes redundant.

Laktamer med formel III er i mange tilfeller kjent eller kan lett fremstilles etter litteraturkjente metoder. Likeledes kjent er i mange tilfelle bromhydrinene II, resp. epoksydene IV (sammenl. her de ovenfor siterte patentskrifter eller J. Med. Chem. 1986, 29, 2194-2201) eller kan fremstilles analogt der angitte metoder. Hittil ikke kjente er slike forbindelser II og IV, hvori R<1> betyr -S0n-Ar med n og Ar med ovennevnte betydning. De kan f.eks. fremstilles etter følgende skjema: Lactams of formula III are in many cases known or can be easily prepared according to methods known in the literature. Likewise known in many cases are the bromohydrins II, resp. the epoxides IV (cf. here the above-cited patent documents or J. Med. Chem. 1986, 29, 2194-2201) or can be prepared analogously to the methods indicated there. So far not known are such compounds II and IV, in which R<1> means -S0n-Ar with n and Ar with the above meaning. They can e.g. produced according to the following scheme:

2,3-dihydro-2H-benzo[b]pyran-4-oner med formel VIII 2,3-dihydro-2H-benzo[b]pyran-4-ones of formula VIII

omsettes med syreklorider Ar-SC^-Cl på i og for seg kjent måte etter typen av Friedel-Crafts-acylering til forbindelser med formel IX hvori R<1> betyr ArSOn (med Ar og n som definert ovenfor) og hvori R<2>, R<3> og R<4> har ovennevnte betydning. Disse forbindelser IX omsettes ved reduksjoner under standardbetingelser, f.eks. med NaBH4 i metanol, til forbindelsene X og deretter underkastes en vannavspaltning, f.eks. med pyridin/fosforoksyklorid, idet det oppstår 2H-benzo[b]pyrener med formel IX are reacted with acid chlorides Ar-SC^-Cl in a manner known per se according to the type of Friedel-Crafts acylation to compounds of formula IX in which R<1> means ArSOn (with Ar and n as defined above) and in which R<2 >, R<3> and R<4> have the above meaning. These compounds IX are reacted by reductions under standard conditions, e.g. with NaBH4 in methanol, to the compounds X and then subjected to a water elimination, e.g. with pyridine/phosphorus oxychloride, resulting in 2H-benzo[b]pyrenes of formula IX

Forbindelsene XI lar seg lett etter standardfremgangsmåter omdanne til epoksydene IV eller bromhydrinene II. Har ved denne reaksjonssekvens R<2> betydningen av NH2 eller OH, så er det eventuelt nødvendig beskyttelsesgrupper, som f.eks. dimetylaminometylengruppen for NH2 eller acetyl- eller metylgrupper for OH-gruppen. Disse avspaltes igjen på egnede trinn, fortrinnsvis etter gjennomføring av de i fremgangsmåt-ene a) eller b) omtalte omsetninger ved vanlige metoder. The compounds XI can easily be converted into the epoxides IV or the bromohydrins II by standard methods. If in this reaction sequence R<2> has the meaning of NH2 or OH, then there are possibly necessary protective groups, such as e.g. the dimethylaminomethylene group for NH2 or acetyl or methyl groups for the OH group. These are split off again in suitable steps, preferably after carrying out the conversions mentioned in procedures a) or b) using normal methods.

Forbindelsene fremstilt ifølge oppfinnelsen med formel I, er som allerede nevnt, antihypertensiva, som kan anvendes som farmasøytika i human- eller veterinærmedisinen. De administ-reres i doseringer på minst 0,001 mg/kg/dag, fortrinnsvis 0,005 mg og spesielt 0,55 mg/kg/dag til maksimalt 10 mg/kg/dag, fortrinnsvis 5 mg/kg/dag og spesielt 2 mg/kg/dag i kapsler, drageer, tabletter, pudder, tapper eller oppløsnin-ger med tilsetninger eller uten tilsetninger av galeniske hjelpestoffer enteralt, f.eks. oralt eller parenteralt (som f.eks. intravenøst ved injeksjon i karsystemet) resp. referert til en vekt på ca. 75 kg. De egner seg til behandling av hypertoni alene eller i kombinasjon med andre hypertensivvirkende legemidler, som f.eks. diuretika, Ca-antagonister eller ACE-hemmere. The compounds produced according to the invention with formula I are, as already mentioned, antihypertensives, which can be used as pharmaceuticals in human or veterinary medicine. They are administered in dosages of at least 0.001 mg/kg/day, preferably 0.005 mg and especially 0.55 mg/kg/day to a maximum of 10 mg/kg/day, preferably 5 mg/kg/day and especially 2 mg/kg /day in capsules, dragees, tablets, powders, drops or solutions with or without additions of galenic excipients enterally, e.g. orally or parenterally (such as intravenously by injection into the vascular system) resp. referred to a weight of approx. 75 kg. They are suitable for the treatment of hypertension alone or in combination with other antihypertensive drugs, such as e.g. diuretics, Ca antagonists or ACE inhibitors.

Disse angivelser refererer seg til et menneske av vekt på 75 kg. These indications refer to a human weighing 75 kg.

Eksempel 1 Example 1

(3R<*>,4S<*>,5'S<*>)-6-cyano-3,4-dihydro-2,2-dimetyl-4-[2'-okso-5'-metyl-l-pyrrolidinyl)-2H-benzo[b]pyran-3-ol og ( 3R*,4S*,5'R* ) - 6-cyano-3 ,4- dihydro-2 ,2- dimetyl- 4- [2 ' -okso-5 ' - metyl-l-pyrrolidinyl)-2H-benzo[b]pyran-3-ol. (3R<*>,4S<*>,5'S<*>)-6-cyano-3,4-dihydro-2,2-dimethyl-4-[2'-oxo-5'-methyl-1-pyrrolidinyl) -2H-benzo[b]pyran-3-ol and (3R*,4S*,5'R*)-6-cyano-3,4-dihydro-2,2-dimethyl-4-[2'-oxo- 5'-methyl-1-pyrrolidinyl)-2H-benzo[b]pyran-3-ol.

Til en oppløsning av 32 g (0,113 mol) 6-cyano-trans-3-bromo-3,4-dihydro-2,2-dimetyl-2H-benzo[b]pyran-4-ol i 200 ml DMSO innføres ved 20°C 3,6 g (0,12 mol) 80^-ig NaH (suspensjon i olje). Etter 1 times omrøring innføres ytterligere 4,8 g To a solution of 32 g (0.113 mol) 6-cyano-trans-3-bromo-3,4-dihydro-2,2-dimethyl-2H-benzo[b]pyran-4-ol in 200 ml of DMSO is introduced at 20 °C 3.6 g (0.12 mol) 80 µg NaH (suspension in oil). After 1 hour of stirring, a further 4.8 g are introduced

(0,16 mol) 80*-ig NaH og 15,9 g (0,16 mol) rasemisk 5-metyl-2-pyrrolidon. Etter 8 timers omrøring ved 20°C helles reaksjonsblandingen i isvann og utfellingen omkrystalliseres fra metanol. (0.16 mol) 80% NaH and 15.9 g (0.16 mol) racemic 5-methyl-2-pyrrolidone. After stirring for 8 hours at 20°C, the reaction mixture is poured into ice water and the precipitate is recrystallized from methanol.

Den omkrystalliserte blanding spaltes over en kiselgelsøyle med CH2C12/CH30H (95:5). (3R*, 4S* , 5 'S* )-6-cyano-3,4-dihydro-2 ,2-dimetyl-4-[2 ' -okso-5 ' -metyl-1 ' -pyrrolidinyl]-2E-benzo-[b]pyran-3-ol isoleres og omkrystalliseres flere ganger fra etanol. The recrystallized mixture is separated over a silica gel column with CH 2 Cl 2 /CH 3 OH (95:5). (3R*, 4S*, 5'S*)-6-cyano-3,4-dihydro-2,2-dimethyl-4-[2'-oxo-5'-methyl-1'-pyrrolidinyl]-2E- benzo-[b]pyran-3-ol is isolated and recrystallized several times from ethanol.

Krystaller av smeltepunkt: 239-240<*>C Crystals of melting point: 239-240<*>C

NMR (d6-DMS0) CH3-signaler S 0,80 (3H, d) NMR (d6-DMS0) CH3 signals S 0.80 (3H, d)

1,17 (3H, s) 1.17 (3H, s)

1,45 (3H, s) 1.45 (3H, s)

Analyse: beregnet for Ci7<H>2g<N>203: 300,37Analysis: calculated for C17<H>2g<N>2O3: 300.37

Beregnet: C, 67,98; H, 6,71; N, 9,33* Calculated: C, 67.98; H, 6.71; N, 9.33*

Funnet: C, 67,9; H, 6,8; N, 9,3* Found: C, 67.9; H, 6.8; N, 9.3*

Den metanolisk moderlut, hvori (3R<*>, 4S<*>, 5 'R** )-6-cyano-3,4-dihydro-2 , 2-dimetyl-4-[2'-okso-5'-metyl-1'-pyrrolidinyl)-2H-benzo[b]pyran-3-ol er anriket, inndampes og forbindelsene isoleres over en kiselsøyle med CH2C12/CH30H (95:5). The methanolic mother liquor, in which (3R<*>, 4S<*>, 5 'R** )-6-cyano-3,4-dihydro-2 , 2-dimethyl-4-[2'-oxo-5'- methyl-1'-pyrrolidinyl)-2H-benzo[b]pyran-3-ol is enriched, evaporated and the compounds isolated over a silica column with CH2C12/CH3OH (95:5).

Krystaller av smeltepunkt: 190-192°C (fra eddikester ) Crystals of melting point: 190-192°C (from acetic acid)

NMR (d6-DMS0) CH3-signaler å 1,20 (3H, s) NMR (d6-DMS0) CH3 signals at 1.20 (3H, s)

1,30 (1,5H, s) 1.30 (1.5H, s)

1,43 (4,5H, s) 1.43 (4.5H, s)

Analyse: Beregnet for Ci7H2qN203<:> 300,37 Analysis: Calculated for Ci7H2qN2O3<:> 300.37

Beregnet: C, 67,98; E, 6,71; N 9,33* Calculated: C, 67.98; E, 6.71; N 9.33*

Funnet: C, 68,0; E, 6,7; N 9,4* Found: C, 68.0; E, 6.7; N 9.4*

Eksempel 2 Example 2

( - )-( 3S ,4R , 5 ' R ) - 6 - cyano-3 ,4-dihydro-2 ,2-d ime ty 1-4-(2 ' -okso-5'-metyl-l-pyrrolidinyl )-2H-benzo[b]pyran-3-ol og (-)- (-)-(3S,4R,5'R)-6-cyano-3,4-dihydro-2,2-dimethyl 1-4-(2'-oxo-5'-methyl-1-pyrrolidinyl) -2H-benzo[b]pyran-3-ol and (-)-

(3R,4S,5 'R )-6-cyano-3 , 4-dihydro-2 , 2-dimetyl-4-(2 '-okso-5 '-metyl-l-pyrrolidinyl )-2H-benzo[b]pyran-3-ol (3R,4S,5'R)-6-cyano-3,4-dihydro-2,2-dimethyl-4-(2'-oxo-5'-methyl-1-pyrrolidinyl)-2H-benzo[b] pyran-3-ol

Til en oppløsning av 30,2 g (0,15 mol) 6-cyano-3,4-dihydro-2,2-dimetyl-3,4-epoksy-2H-benzo[b]pyran i 150 ml dimetylsulfoksyd innføres 6 g (0,2 mol) 80*-ig NaH og 15 g (0,15 mol) To a solution of 30.2 g (0.15 mol) 6-cyano-3,4-dihydro-2,2-dimethyl-3,4-epoxy-2H-benzo[b]pyran in 150 ml of dimethylsulfoxide, 6 g (0.2 mol) 80*-ig NaH and 15 g (0.15 mol)

(R)-5-metyl-2-pyrrolidon (se René Amstutz, Bjørn Ringdahl, Bo Karlén, Margareth Rock og Donald J. Jenden, J. Med. Chem. 1985, 28, 1760-1765) og omrøres i 6 timer ved 20°C. Reaksjonsblandingen innføres i isvann, utfellingen vaskes nøytral, tørkes og adskilles over en kiselgelsøyle med CH2CI2/CH3OH 19:1 og omkrystalliseres fra etanol. Man får i første rekke (-)-(3R,4S,5'R)-6-cyano-3,4-dihydro-2,2-dimetyl-4-(2'-okso-5'-metyl-1'-pyrrolidinyl)-2H-benzo[b]pyran-3-ol. (R)-5-methyl-2-pyrrolidone (see René Amstutz, Bjørn Ringdahl, Bo Karlén, Margareth Rock and Donald J. Jenden, J. Med. Chem. 1985, 28, 1760-1765) and stirred for 6 hours at 20°C. The reaction mixture is introduced into ice water, the precipitate is washed neutral, dried and separated over a silica gel column with CH2CI2/CH3OH 19:1 and recrystallized from ethanol. One obtains primarily (-)-(3R,4S,5'R)-6-cyano-3,4-dihydro-2,2-dimethyl-4-(2'-oxo-5'-methyl-1' -pyrrolidinyl)-2H-benzo[b]pyran-3-ol.

Krystaller av smeltepunkt: 184"C a<20>D = -27,8" c = 1 i etanol. Crystals of melting point: 184"C a<20>D = -27.8" c = 1 in ethanol.

Som det andre langsommere eluerende produkt får man etter ekstra krystallisering fra etanol (-)-(3S,4R,5'R)-6-cyano-3,4-dihydro-2,2-dimetyl-4-(2'-okso-5'-metyl-l-pyrrolidinyl)-2H-benzo[b]pyran-3-ol. As the second slower eluting product, after additional crystallization from ethanol (-)-(3S,4R,5'R)-6-cyano-3,4-dihydro-2,2-dimethyl-4-(2'-oxo) is obtained -5'-methyl-1-pyrrolidinyl)-2H-benzo[b]pyran-3-ol.

Krystaller av smeltepunkt: 258°C oc20j) = -86° (c= 1, etanol) Crystals of melting point: 258°C oc20j) = -86° (c= 1, ethanol)

Eksempel 3 Example 3

6-cyano-3,4-dihydro-2,2-dimetyl-trans-4-(2'-okso-3'-metyl-l-pyrrolidinyl )-2H-benzo[b]pyran-3-ol 6-cyano-3,4-dihydro-2,2-dimethyl-trans-4-(2'-oxo-3'-methyl-1-pyrrolidinyl)-2H-benzo[b]pyran-3-ol

Diastereomer A .og diastereomer B Diastereomer A .and diastereomer B

Diastereomer B: Diastereomer B:

Til en oppløsning av 16 g (0,056 mol) 6-cyano-trans-3-bromo-3,4-dihydro-2,2-dimetyl-2H-benzo[b]pyran-4-ol i 80 ml dimetylsulfoksyd settes 1,8 g (0,06 mol) 80*-ig NaH. Etter 1 times omrøring ved 20°C tilsettes ytterligere 2,4 g (0,08 mol) 80*-ig NaH og 8,6 g (0,084 mol) rasemisk 3-metyl-2-pyrrolidon og deretter omrøres i 16 timer ved 20°C. Etter avkjøling og inndrypping av 160 ml vann, frasuges utfellingen, vaskes med litt vann og tørkes. To a solution of 16 g (0.056 mol) 6-cyano-trans-3-bromo-3,4-dihydro-2,2-dimethyl-2H-benzo[b]pyran-4-ol in 80 ml of dimethyl sulfoxide is added 1, 8 g (0.06 mol) 80*-ig NaH. After stirring for 1 hour at 20°C, a further 2.4 g (0.08 mol) of 80% NaH and 8.6 g (0.084 mol) of racemic 3-methyl-2-pyrrolidone are added and then stirred for 16 hours at 20 °C. After cooling and instilling 160 ml of water, the precipitate is suctioned off, washed with a little water and dried.

Råproduktet omkrystalliseres to ganger fra eddiketylester/- metanol og deretter fra dimetylformamid/metanol. Den diastereomere lar seg således renisolere. The crude product is recrystallized twice from ethyl acetate/methanol and then from dimethylformamide/methanol. The diastereomer can thus be reisolated.

Krystaller av smeltepunkt: 261-263°C Crystals of melting point: 261-263°C

NMR (d6-DMS0) CH3-signaler: S 1,19 (3H, s) NMR (d 6 -DMSO) CH 3 signals: S 1.19 (3H, s)

1,21 (6H, s) 1.21 (6H, s)

1,45 (3H, s) 1.45 (3H, s)

Diastereomer A: Diastereomer A:

Moderluten fra ovennevnte forsøk inndampes og resten kromatograferes over en kiselgelsøyle med eddikester/petroleter (9:1). Diastereomeren A adskilles og omkrystalliseres fra etanol. The mother liquor from the above experiment is evaporated and the residue is chromatographed over a silica gel column with vinegar/petroleum ether (9:1). The diastereomer A is separated and recrystallized from ethanol.

Krystaller av smeltepunkt: 210-211°C Crystals of melting point: 210-211°C

NMR (d6-DMS0) CH3-signaler: S 1,13 (3H, d) NMR (d6-DMSO) CH3 signals: S 1.13 (3H, d)

1,20 (3H, s) 1.20 (3H, s)

1,47 (3H, s) 1.47 (3H, s)

Eksempel 4 Example 4

(3R<*>,4S<*>,5'S<*>)-3,4-dihydro-2,2-dimetyl-6-metylsulfonyl-4-(2'-okso-5'-metyl-l-pyrrolidinyl)-2H-benzo[b]pyran-3-ol (3R<*>,4S<*>,5'S<*>)-3,4-dihydro-2,2-dimethyl-6-methylsulfonyl-4-(2'-oxo-5'-methyl-1-pyrrolidinyl) -2H-benzo[b]pyran-3-ol

Til en oppløsning av 10,2 g (0,04 mol) 3,4-dihydro-2,2-dimetyl-3,4-epoksy-6-metylsulfonyl-2H-benzo[b]pyran og 4 g (0,04 mol) rasemisk 5-metyl-2-pyrrolidon i 50 ml DMSO innføres ved 20°C 1,5 g (0,05 mol) 80*-ig NaH. Man oppvarmer i 30 minutter ved 45° C, lar det omrøre 5 timer ved 20° C og heller i isvann. Det klares med aktivkull og produktet utsaltes. Utfellingen frasuges og diastereomeren A adskilles over en kiselgelsøyle med etanol/eddikester/toluen/petroleter 2:2:1:1 og omkrystalliseres fra eddikester. To a solution of 10.2 g (0.04 mol) 3,4-dihydro-2,2-dimethyl-3,4-epoxy-6-methylsulfonyl-2H-benzo[b]pyran and 4 g (0.04 mol) racemic 5-methyl-2-pyrrolidone in 50 ml DMSO is introduced at 20°C 1.5 g (0.05 mol) 80*-ig NaH. It is heated for 30 minutes at 45° C, left to stir for 5 hours at 20° C and poured into ice water. This is done with activated charcoal and the product is salted out. The precipitate is suctioned off and the diastereomer A is separated over a silica gel column with ethanol/acetic ester/toluene/petroleum ether 2:2:1:1 and recrystallized from ethyl acetate.

Krystaller av smeltepunkt: 190-193°C Crystals of melting point: 190-193°C

NMR (d6-DMSO) CH3-signaler: S 0,74 (3H, d) NMR (d6-DMSO) CH3 signals: S 0.74 (3H, d)

1,18 (3H, s) 1.18 (3H, s)

1,47 (3H, s) 1.47 (3H, s)

Fremstilling av utgangsmaterialer: p-metylsulfonylfenyl-1,1-dimetylpropargyleter får man fra p-metylsulfonylfenol og 3-metyl-3-klorbutin på i og for seg kjent måte. Preparation of starting materials: p-methylsulfonylphenyl-1,1-dimethylpropargyl ether is obtained from p-methylsulfonylphenol and 3-methyl-3-chlorobutene in a manner known per se.

Smeltepunkt: 64-65°C (fra litt petroleter) Melting point: 64-65°C (from a little petroleum ether)

2,2-dimetyl-6-metylsulfonylkromen får man ved oppvarming av p-metylsulfonylfenyl-1,1-dimetylpropargyleter i 1,2-diklor-benzen ved 180"C. 2,2-Dimethyl-6-methylsulfonyl chrome is obtained by heating p-methylsulfonylphenyl-1,1-dimethylpropargyl ether in 1,2-dichlorobenzene at 180°C.

Smeltepunkt: 107-108"C (fra diisopropyleter) Melting point: 107-108"C (from diisopropyl ether)

3-brom-3,4-dihydro-2,2-dimetyl-6-metylsul f ony1-2H-benzo-[b]pyran-4-ol får man fra 2,2-dimetyl-6-metylsulfonylkromen og N-bromsuksinimid i en 9:1 dimetylsulfoksyd/B^O-blanding. 3-bromo-3,4-dihydro-2,2-dimethyl-6-methylsulfonyl-2H-benzo-[b]pyran-4-ol is obtained from 2,2-dimethyl-6-methylsulfonyl chrome and N-bromosuccinimide in a 9:1 dimethyl sulfoxide/B2O mixt.

Smeltepunkt: 140-141°C fra diisopropyleter. Melting point: 140-141°C from diisopropyl ether.

3,4-dihydro-2,2-dimetyl-3, 4 - epoksy-6-metyl sul f onyl -2H-benzo[b]pyran-4-ol får man ved omsetning av 3-brom-3,4-dihydro-2,2-dimetyl-6-metylsulfonyl-2H-benzo[b]pyran-4-ol med natriumhydrid i DMSO. 3,4-dihydro-2,2-dimethyl-3,4-epoxy-6-methylsulfonyl-2H-benzo[b]pyran-4-ol is obtained by reacting 3-bromo-3,4-dihydro -2,2-dimethyl-6-methylsulfonyl-2H-benzo[b]pyran-4-ol with sodium hydride in DMSO.

Smeltepunkt: 1'65 °C (fra eddikester). Melting point: 1'65 °C (from acetic acid).

Eksempel 5 Example 5

(3R*,4S*,5'S*)-2 , 2-dimetyl-7-metoksy-6-fenylsulfonyl-4-( 2 ' - okso-5'-metyl-1'-pyrrolidinyl)-2H-benzo[b]pyran-3-ol og (3R*,4S*,5'R*)-2 ,2-dimetyl-7-metoksy-6-fenylsulfonyl-4-(2'-okso-5'-metyl-1'-pyrroiidinyl)-2H-benzo[b]pyran-3-ol (3R*,4S*,5'S*)-2,2-dimethyl-7-methoxy-6-phenylsulfonyl-4-(2'-oxo-5'-methyl-1'-pyrrolidinyl)-2H-benzo[b] pyran-3-ol and (3R*,4S*,5'R*)-2,2-dimethyl-7-methoxy-6-phenylsulfonyl-4-(2'-oxo-5'-methyl-1'-pyrroiidinyl )-2H-benzo[b]pyran-3-ol

Til en oppløsning av 9,6 g 3-brom-3,4-dihydro-2,2-dimetyl-7-metoksy-6-fenylsulfonyl-2H-benzo[b]pyran-4-ol i 60 ml dimetylsulfoksyd setter man 1,62 g (0,0675 mol) NaH i form av en 80#-ig suspensjon samt 10,3 g (0,11 mol) (± )-5-metyl-2-pyrrolidinon og omrører blandingen 4 timer ved 40'C. Deretter heller man blandinger på isvann og frasuges. Resten vaskes nøytralt, tørkes og underkastes kromatografl på kiselgel med elueringsmiddel toluen/metylenklorid/metanol 10:1:1. Som første hovedprodukt elueres derved (3R<*>,4S<*>,-5 'R* )-2 , 2-dimetyl-7-metoksy-6-fenylsulfonyl-4-(2 '-okso-5'-metyl-1'-pyrrolidinyl )-2H-benzo[b]pyran-3-ol: smeltepunkt 214-215°C, NMR (d6-DMS0) S (ppm): 1,17, 1,27, 1,4 (s,s,s; 9H, 2,2-dimetyl og 5-metyl). 1 .62 g (0.0675 mol) NaH in the form of an 80# suspension as well as 10.3 g (0.11 mol) (± )-5-methyl-2-pyrrolidinone and stir the mixture for 4 hours at 40°C . Mixtures are then poured into ice water and suctioned off. The residue is washed neutrally, dried and subjected to chromatography on silica gel with eluent toluene/methylene chloride/methanol 10:1:1. As the first main product, (3R<*>,4S<*>,-5 'R* )-2, 2-dimethyl-7-methoxy-6-phenylsulfonyl-4-(2 '-oxo-5'-methyl- 1'-pyrrolidinyl)-2H-benzo[b]pyran-3-ol: mp 214-215°C, NMR (d6-DMS0) S (ppm): 1.17, 1.27, 1.4 (s, s,s; 9H, 2,2-dimethyl and 5-methyl).

Som annet hovedprodukt får man det langsommere eluerende (3R* ,4S* ,5 'S* )-2 , 2-dimetyl-7-metoksy-6-fenylsulfonyl-4-(2'-okso-5'-metyl-1'-pyrrolidinyl)-2h-benzo[b]pyran-3-ol. Smeltepunkt: 244-245°C; NMR (d6-DMS0) S (ppm): 0,73 (d, 3H, 5'-CH3). As another main product, the slower eluting (3R* ,4S* ,5 'S* )-2 , 2-dimethyl-7-methoxy-6-phenylsulfonyl-4-(2'-oxo-5'-methyl-1') is obtained -pyrrolidinyl)-2h-benzo[b]pyran-3-ol. Melting point: 244-245°C; NMR (d 6 -DMSO) S (ppm): 0.73 (d, 3H, 5'-CH 3 ).

Fremstilling av utgangsmateriale: 3-brom-3 ,4-dihydro-2,2-dimetyl-7-metoksy-6-fenylsulfonyl-2H-benzo[b]pyran-4-ol får man av 2,2-dimetyl-7-metoksy-6-fenylsulfonyl-2H-kromen og N-bromsuksinimid i en 9:1 dimetylsulfoksyd/H20-blanding. Smeltepunkt: 202-203°C. Preparation of starting material: 3-bromo-3,4-dihydro-2,2-dimethyl-7-methoxy-6-phenylsulfonyl-2H-benzo[b]pyran-4-ol is obtained from 2,2-dimethyl-7- methoxy-6-phenylsulfonyl-2H-chrome and N-bromosuccinimide in a 9:1 dimethylsulfoxide/H 2 O mixt. Melting point: 202-203°C.

2,2-dimetyl-7-metoksy-6-fenylsulfonyl-2H-kromen får man av 2,2-dimetyl-4-hydroksy-7-metoksy-6-fenylsulfonyl-kroman med pyridin/fosforoksyklorid i benzen. 2,2-Dimethyl-7-methoxy-6-phenylsulfonyl-2H-chrome is obtained from 2,2-dimethyl-4-hydroxy-7-methoxy-6-phenylsulfonyl chromane with pyridine/phosphorus oxychloride in benzene.

Smeltepunkt: 140-141°C. Melting point: 140-141°C.

2,2-dimetyl-4-hydroksy-7-metoksy-6-fenylsulfonylkromam får man av 2,2-dimetyl-7-metoksy-6-fenylsulfonyl-kroman-4-on med natriumborhydrid i metanol. 2,2-dimethyl-4-hydroxy-7-methoxy-6-phenylsulfonylchromam is obtained from 2,2-dimethyl-7-methoxy-6-phenylsulfonyl-chroman-4-one with sodium borohydride in methanol.

Smeltepunkt: 146-147°C. Melting point: 146-147°C.

2,2-dimetyl-7-metoksy-6-fenylsulfonylkroman-4-on får man av fenylsulfonylklorid, 2,2-dimetyl-7-metoksykroman-4-on og aluminiumklorid i metylenklorid. 2,2-dimethyl-7-methoxy-6-phenylsulfonylchroman-4-one is obtained from phenylsulfonyl chloride, 2,2-dimethyl-7-methoxychroman-4-one and aluminum chloride in methylene chloride.

Smeltepunkt: 223-225°C. Melting point: 223-225°C.

Eksempel 6 Example 6

(3R*,4S*,5'S<*>)-3,4-dihydro-2,2-dimetyl-6-fenylsulfonyl-4-[2'-okso-5 * -metyl-l-pyrrolidinyl-2H-benzo[b]pyran-3-ol og (3R* , 4S* , 5 'R** )-3,4-dihydro-2,2-dimetyl-6-fenylsulfonyl-4-(2'-okso-5'-metyl-l-pyrrolidinyl)-2H-benzo[b]pyran-3-ol. (3R*,4S*,5'S<*>)-3,4-dihydro-2,2-dimethyl-6-phenylsulfonyl-4-[2'-oxo-5*-methyl-1-pyrrolidinyl-2H-benzo[ b]pyran-3-ol and (3R* , 4S* , 5 'R** )-3,4-dihydro-2,2-dimethyl-6-phenylsulfonyl-4-(2'-oxo-5'-methyl -1-pyrrolidinyl)-2H-benzo[b]pyran-3-ol.

Forbindelsene fremstilles analogt eksempel 5 med rasemisk 5-metylpyrrolidin-2-on. Deretter spaltes blandingen på en kiselgelsøyle med metylenklorid/eddikester/etanol (94:3:3). The compounds are prepared analogously to example 5 with racemic 5-methylpyrrolidin-2-one. The mixture is then separated on a silica gel column with methylene chloride/acetic acid/ethanol (94:3:3).

(3R* ,4S* ,5 'S* )-3,4-dihydro-2,2-dimetyl-6-fenylsulfonyl-4-(2'-okso-5'-metyl-l-pyrroldinyl)-2H-benzo[b]pyran-3-ol. (3R* ,4S* ,5 'S* )-3,4-dihydro-2,2-dimethyl-6-phenylsulfonyl-4-(2'-oxo-5'-methyl-1-pyrroldinyl)-2H-benzo [b]pyran-3-ol.

Krystaller av smeltepunkt: 216-217°C fra etanol. Crystals of melting point: 216-217°C from ethanol.

NMR (d6-DMS0) 5'-CH3-signal: S 0,48 (3H, pseudoduplett). NMR (d 6 -DMSO) 5'-CH 3 signal: S 0.48 (3H, pseudo-doublet).

(3R*,4S<*>,5'R<*>)-3,4-dihydro-2,2-dimetyl-6-fenylsulfonyl-4-(2'-okso-5'-metyl-l-pyrrolidinyl )-2H-benzo[b]pyran-3-ol. (3R*,4S<*>,5'R<*>)-3,4-dihydro-2,2-dimethyl-6-phenylsulfonyl-4-(2'-oxo-5'-methyl-1-pyrrolidinyl) -2H-benzo[b]pyran-3-ol.

Krystaller av smeltepunkt: 240-241°C fra DMF/etanol. Crystals of melting point: 240-241°C from DMF/ethanol.

NMR (dfc-DMSO) 5'-CH3-signal: 5 1,27 (3H, d). NMR (dfc-DMSO) 5'-CH 3 signal: δ 1.27 (3H, d).

Farmakologiske data Pharmacological data

Innvirkningen av forbindelsene på det arterielle blodtrykk undersøkes på narkotiserte normotensive hann Sprague-Dawley rotter. Som bedøvelsesmiddel ble det anvendt "Nembutal" i en dosering på 70 mg/kg i.p. Blodtrykksmålingen foregikk ved hjelp av kateter i Arteria carotis med en trykkomdanner (Statham P 23 Db). For i.v. applikasjon ble stoffene oppløst i dimetylsulfoksyd og fortynnet med vann. Konsentrasjonen av stammoppløsningen var 1 mg/ml, idet oppløsningen inneholdt 15* metylsulfoksyd. Deler av denne stammoppløsning ble fortynnet for injeksjon resp. friskt med vann. The effect of the compounds on the arterial blood pressure is investigated in anesthetized normotensive male Sprague-Dawley rats. "Nembutal" was used as anesthetic in a dosage of 70 mg/kg i.p. Blood pressure was measured using a catheter in the carotid artery with a pressure transducer (Statham P 23 Db). For i.v. application, the substances were dissolved in dimethyl sulphoxide and diluted with water. The concentration of the stock solution was 1 mg/ml, the solution containing 15* methyl sulphoxide. Parts of this stock solution were diluted for injection or fresh with water.

Det kom til anvendelse konsentrasjoner på 50 (100 ), 30 (30 ) og 10 (10) >jg/ml for i.v. applikasjon, 100 (100 ), 50 (60 ) og 30 (30) jjg/ml for i.d. applikasjon. De angitte tall refererer seg derved til eksperiment a), de i klammer angitte tall til eksperiment b). Concentrations of 50 (100 ), 30 (30 ) and 10 (10) µg/ml were used for i.v. application, 100 (100 ), 50 (60 ) and 30 (30) jjg/ml for i.d. application. The indicated numbers thereby refer to experiment a), the numbers indicated in brackets refer to experiment b).

Blodtrykksmålingen ble foretatt etter 0; 0,2; 0,4; 1; 2; 3; 10; 15 og 30 minutter etter i.v. applikasjon, samt etter 0; 1; 3; 5; 10; 15; 20; 30; 40; 50 og 60 minutter etter i.d. applikasjon. EE>25 verdiene ble bestemt fra minima av de resp. blodtrykkskurver. The blood pressure measurement was taken after 0; 0.2; 0.4; 1; 2; 3; 10; 15 and 30 minutes after i.v. application, as well as after 0; 1; 3; 5; 10; 15; 20; 30; 40; 50 and 60 minutes after i.d. application. The EE>25 values were determined from the minima of the resp. blood pressure curves.

a) (±)-(3R*,4S*,5'S* )-6-cyano-3,4-dihydro-2,2-dimetyl-4-(2'-okso-5'-metyl-1'-pyrrolidinyl)-2H-benzo[b]pyran-3-ol a) (±)-(3R*,4S*,5'S* )-6-cyano-3,4-dihydro-2,2-dimethyl-4-(2'-oxo-5'-methyl-1'-pyrrolidinyl )-2H-benzo[b]pyran-3-ol

(forbindelse fra eksempel 1): (connection from example 1):

ED25 = 19 jJg/kg (i.v.) ED25 = 42 pg/kg (i.d.). ED25 = 19 µg/kg (i.v.) ED25 = 42 pg/kg (i.d.).

b ) (±)-6-cyano-3,4-dihydro-2,2-dimetyl-trans-4-(2-okso-l-pyrrolidinyl-2H-benzo[b]pyran-3-ol b ) (±)-6-cyano-3,4-dihydro-2,2-dimethyl-trans-4-(2-oxo-1-pyrrolidinyl-2H-benzo[b]pyran-3-ol

(eksempel fra J. Med. Chem. 1986, 29, 2194-2201). (example from J. Med. Chem. 1986, 29, 2194-2201).

E<D>25 = 42 pg/kg (i.v.) ED25 <=> 82 jjg/kg (i.d.). E<D>25 = 42 pg/kg (i.v.) ED25 <=> 82 jjg/kg (i.d.).

Påvirkning av KCl-indusert rytmisk kontraksjoner på marsvin-uretere in vitro Influence of KCl-induced rhythmic contractions on guinea pig ureters in vitro

Metode: Method:

Hannmarsvin ble avlivet ved nakkeslag og avblødning fra Carotidene. De to uretere ble med en gang uttatt og området nær nyrekjeglen ble unngått p.g.a. den der skrittgjørings-aktivitet. 2 cm lange stykker ble i første rekke i en petriskål, som inneholdt Tyrode-oppløsning, befridd for bindevev og deretter resp. hengt med en forspenning på 4,9 mN (=* 0,5 p) i et 25 ml organbad (Fa. Rhema Labortechnik, Hofheim). Organbadet inneholdt en med 37°C holdt og med Carbogen (95* 02, 5% C02j ) gjennomperlet Tyrodeoppløsning med følgende sammensetning (mmol/1): NaCl 137, KC1 2,68, MgS04 1,05, CaCl2 1,8, NaH2P04 0,41, NaEC03 11,9, glukose 5,55. Male guinea pigs were euthanized by neck blow and bleeding from the carotids. The two ureters were removed at once and the area near the renal cone was avoided due to that pacing activity. 2 cm long pieces were first in a Petri dish, which contained Tyrode solution, freed of connective tissue and then resp. suspended with a pretension of 4.9 mN (=* 0.5 p) in a 25 ml organ bath (Fa. Rhema Labortechnik, Hofheim). The organ bath contained a Tyrode solution kept at 37°C and permeated with Carbogen (95*02, 5% C02j ) with the following composition (mmol/1): NaCl 137, KC1 2.68, MgS04 1.05, CaCl2 1.8, NaH2P04 0.41, NaEC03 11.9, glucose 5.55.

Kontraksjonene ble målt isometrisk under anvendelse av Gould/Statham UC2 givere. Etter en ekvilibreringstid på minst 15 minutter ble det til organbadet satt KC1 idet det ble oppmålt en konsentrasjon på 4xl0~<2> mol/l. Agonisten ble hensatt 2 minutter i badet, idet det opptrådte faslske kontraksjoner, uten å bevirke nevneverdig økning av grunn-spenningen. Contractions were measured isometrically using Gould/Statham UC2 transducers. After an equilibration time of at least 15 minutes, KC1 was added to the organ bath, with a concentration of 4x10~<2> mol/l being measured. The agonist was left in the bath for 2 minutes, as phasic contractions occurred, without causing a significant increase in the baseline tension.

Deretter ble det utspylt 1 minutt, hvorpå de rytmiske kontraksjoner med en gang opphørte. Etter en annen passering av agonistinngivning av spyleprosess ble prøvestoffet (Benzopyranderivatet) tilsatt i organbadet (i form av en oppløsning på 0,1 ml etanol, som utgjorde sluttkonsentra-sjoner i alle tilfeller IO-'7 mol/l) og avventet en inn-virkningstid på 1 minutt før KC1 ble tilsatt. På den følgende skylleprosess knyttet det seg to avsluttende agonisttilsetninger/spyleprosesser. Av de resp. 2 minutters perioder under KCl-innvirkning ble det bestemt følgende parametere: 1. midlere kontraksjonskraft, 2. kontraksjone-nes frekvens og 3. produkt av midlere kontraksjonskraft og f rekvens. It was then flushed out for 1 minute, after which the rhythmic contractions immediately ceased. After another passage of agonist administration through the flushing process, the test substance (the benzopyran derivative) was added to the organ bath (in the form of a solution of 0.1 ml of ethanol, which amounted to final concentrations in all cases of 10-7 mol/l) and awaited an in- action time of 1 minute before KC1 was added. In the following rinsing process, two final agonist additions/rinsing processes were connected. Of the resp. During 2 minute periods under KCl exposure, the following parameters were determined: 1. mean contraction force, 2. frequency of contractions and 3. product of mean contraction force and f frequency.

Utelukkelseskriterier var midlere kontraksjonskrefter under 4 mN eller frekvenser under 2/min. ved mer enn en av de fire perioder hvor det bare i badet var tilstede agonist KC1. Vurdert ble den prosentuelle hemming under prøvestoffet sammenlignet med den midlere verdi av to forløp. Exclusion criteria were mean contraction forces below 4 mN or frequencies below 2/min. at more than one of the four periods where agonist KC1 was present only in the bath. The percentage inhibition under the test substance was evaluated compared to the mean value of two runs.

Ved siden av den aritmetiske middelverdi (x) ble det beregnet standardfeil av middelverdien (SEM). Next to the arithmetic mean (x), the standard error of the mean (SEM) was calculated.

Resultater Results

Litteratur: Literature:

P. Schiantarelli og W.Murmann, P. Schiantarelli and W. Murmann,

Antispasmodic Activity of Rociverine on the Smooth Musculature of the Urinary Tract. Antispasmodic Activity of Rociverine on the Smooth Musculature of the Urinary Tract.

Arzneim.-Forsch./Drug Res. 30, 1102-1109 (1980) Arzneim.-Forsch./Drug Res. 30, 1102-1109 (1980)

Claims

1. Analogous process for the preparation of therapeutically active N-benzopyranyl lactams of formula I in which R<1> means CN, SO2-(C1-Cb)-alkyl or SO2-Ph, R<2> means H, (C1-C2)-Alkoxy, R<3> and R<4> are the same and mean alkyl with 1-4 C atoms, X means a chain (CH 2 )m, which is substituted by a (C 2 -C 2 )- alkyl group and m means 3 or 4, with the OH group in the 3-position and the lactam group in the 4-position always having a trans configuration, characterized by) compounds of formula II in which R<1>, R2", R3 and R<4> have the above meaning, are reacted with lactams of formula III or b) compounds of formula IV in which R<1>, R<2>, R<3> and R<4> have the above meaning, is reacted with lactams of formula III.

2. Analogous process according to claim 1, for the preparation of (3R*,4S*,5'S*)-6-cyano-3,4-dihydro-2,2-dimethyl-4-[2'-oxo-5'-methyl-1- pyrrolidinyl)-2H-benzo[b]pyran-3-ol, characterized by using correspondingly substituted starting materials.

3. Analogous method according to claim 1, for the preparation of (3R*,4S*,5'R*)-6-cyano-3,4-dihydro-2,2-dimethyl-4-[2'-oxo-5'-methyl- 1-pyrrolidinyl)-2H-benzo[b]pyran-3-ol, characterized in that similarly substituted starting materials are used.

4. Analogous process according to claim 1, for the production of (-)-(3S,4R,5*R)-6-cyano-3,4-dihydro-2,2-dimethyl-4-(2'-oxo-5'-methyl) -1-pyrrolidinyl)-2H-benzo[b]pyran-3-ol, characterized in that similarly substituted starting materials are used.

5. Analogous process according to claim 1, for the preparation of (-)-(3R,4S,5*R)-6-cyano-3,4-dihydro-2,2-dimethyl-4-(2'-oxo-5'-methyl) -1-pyrrolidinyl)-2H-benzo[b]pyran-3-ol, characterized in that similarly substituted starting materials are used.

6. Analogous process according to claim 1, for the preparation of 6-cyano-3,4-dihydro-2,2-dimethyl-trans-4-(2'-oxo-3'-methyl-1-pyrrolidinyl)-2H-benzo [b] pyran-3-ol, characterized by using correspondingly substituted starting materials.

7. Analogous process according to claim 1 for the preparation of (3R<*>,4S<*>,5'S<*>)-3,4-dihydro-2,2-dimethyl-6-methylsulfonyl-4-(2'-oxo-5' -methyl-1-pyrrolidinyl)-2H-benzo[b]pyran-3-ol, characterized by using correspondingly substituted starting materials.

8. Analogous method according to claim 1 for the production of (3R*,4S*,5'S*)-2,2-dimethyl-7-methoxy-6-phenylsulfonyl-4-(2'-oxo-5'-methyl-1'- pyrrolidinyl)-2H-benzo[b]pyran-3-ol, characterized by using correspondingly substituted starting materials.

9. Analogous method according to claim 1, for the preparation of (3R*,4S*,5'R*)-2,2-dimethyl-7-methoxy-6-phenylsulfonyl-4-(2'-oxo-5'-methyl-1' -pyrrolidinyl)-2H-benzo[b]pyran-3-ol, characterized by using correspondingly substituted starting materials.