DK145699B - METHOD OF ANALOGUE FOR PREPARING PENICILLAL COMPOUNDS - Google Patents

Description

/ ® l (19) DANMARK \ p.a// ® l (19) DENMARK \ p.a /

rø (12) FREMLÆGGELSESSKRIFT <n> 145699 Brow (12) PUBLICATION <n> 145699 B

DIREKTORATET FOR PATENT- OG VAREMÆRKEVÆSENETDIRECTORATE OF THE PATENT AND TRADEMARKET SYSTEM

(21) Ansøgning nr. 1197/72 (51) |nt.CI.3 C 07 D 499/32 (22) Indleveringsdag 15· tnar. 1972 (24) Løbedag 15· tnar. 1972 (41) Aim. tilgængelig 14. sep. 1975 (44) Fremlagt 51 · jan. 1983 (86) International ansøgning nr. -(86) International indleveringsdag -(85) Videreførelsesdag -(62) Stamansøgning nr. -(21) Application No. 1197/72 (51) | nt.CI.3 C 07 D 499/32 (22) Filing day 15 · tnar. 1972 (24) Race day 15 · tnar. 1972 (41) Aim. available Sep 14 1975 (44) Presented 51 · Jan. 1983 (86) International application # - (86) International filing day - (85) Continuation day - (62) Master application no -

(30) Prioritet 15· mar. 1972, 11689/72, GB(30) Priority 15 Mar 1972, 11689/72, GB

(71) Ansøger ASTRA LAEKEMEDEL AKTIEBOLAG, 151 85 Soedertaelje, SE.(71) Applicant ASTRA LEKEMEDEL SHARE COMPANY, 151 85 Soedertaelje, SE.

(72) Opfinder Bertil Åke Ekstroem, SE: Berndt Olof Harald _Sjoeberg, SE:(72) Inventor Bertil Åke Ekstroem, SE: Berndt Olof Harald _Sjoeberg, SE:

Peter Bamberg, SE.Peter Bamberg, SE.

(74) Fuldmægtig Plougmann & Vingtoft Patentbureau.(74) Plougmann & Vingtoft Patent Bureau.

(54) Analogifremgangsmåde til fremstil* ling af penicillinforbindelser.(54) Analogous process for the preparation of penicillin compounds.

Den foreliggende opfindelse angår en analogifremgangsmåde til fremstilling af hidtil ukendte penicillansyrederivater, nemlig estere af amidinopenicillansyrer med den almene formel IThe present invention relates to an analogous process for the preparation of novel penicillanic acid derivatives, namely esters of amidinopenicillanic acids of general formula I

/'"Λ sx /0¾/ '"Λ sx / 0¾

m (CH,) N - CH = H - CH - CH C- _ Im (CH,) N - CH = H - CH - CH C - _ I

OT I 4 LD CO - N-CH—COC—CH-O-COO-R*OT I 4 LD CO - N-CH-COC-CH-O-COO-R *

-O-ISLAND

\— eller terapeutisk acceptable syreadditionssalte deraf, i hvilken for-. 4 3C mel n er et tal 3-7; R betegner alkyl med 1-8 carbonatomer; Q c og R betegner hydrogen eller methyl.Or therapeutically acceptable acid addition salts thereof, in which 4 3C flour n is a number 3-7; R is alkyl of 1-8 carbon atoms; Q c and R represent hydrogen or methyl.

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Som eksempler på radikaler, der omfattes af de ovenfor anførte definitioner og af definitionerne i hele nærværende beskrivelse, kan nævnes: alkyl: methyl, ethyl, propyl, isopropyl, butyl,' isobutyl, pentyl, hexyl, heptyl, octyl og 2-ethyl-hexyl; ch CH2 - CH2 ch2 - CH2 - /CH2 “ N\ “ N\ XIH£ XH2 - CH2 XCH2 - CH2 ch2 - ch2 - ch2 - *^>.Ch2 ^^-CH2 -Examples of radicals encompassed by the above definitions and the definitions throughout this specification may be mentioned: alkyl: methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, hexyl, heptyl, octyl and 2-ethyl hexyl; ch CH2 - CH2 ch2 - CH2 - / CH2 “N \” N \ XIH £ XH2 - CH2 XCH2 - CH2 ch2 - ch2 - ch2 - * ^> .Ch2 ^^ - CH2 -

De ovenfor angivne illustrerende eksempler illustrerer radikalerne R4 og K5 samt andre radikaler, der omtales i nærværende beskrivelse, inden for den for hvert radikal angivne definition og inden for de grænser med hensyn til carbonatomantal, som måtte være anført for hvert enkelt radikal.The illustrative examples given above illustrate the radicals R4 and K5, as well as other radicals referred to in this specification, within the definition given for each radical and within the limits of carbon atom numbers which may be listed for each radical.

De ifølge den foreliggende opfindelse fremstillede forbindelser er værdifulde ved behandlingen af infektionssygdomme i mennesker eller dyr forårsaget af bakterielle organismer. De kan isoleres og anvendes som sådanne, men også, afhængigt af tilstedeværelsen af basiske eller svire grupper i molekylet, i form af salte med farmaceutisk tolerable organiske eller uorganiske syrer eller baser. Eksempler på velegnede syrer er saltsyre, brombrintesyre, svovlsyre, phosphor-syre, eddikesyre, vinsyre, citronsyre og fumarsyre. Eksempler på egnede baser er natriumhydroxid, kaliumhydroxid, calciumhydroxid, aluminiumhydroxid, ammoniumhydroxid, ikke-toxiske aminer såsom tri-alkylaminer, hervinder triethylamin, procain, dibenzylamin, N-benzyl--β-phenylethylamin, 1-ephenamin, Ν,κΡ"-dibenzylethylendiamin, dehydro-abietylamin, N,N^-bis-dehydroabietylethylendiamin, N-(lavere)alkyl-pvpe-ridin (f.eks. N-ethyl-piperidin) og andre baser, som er blevet anvendt til fremstilling af salte med penicilliner, 3 145899The compounds of the present invention are valuable in the treatment of infectious diseases in humans or animals caused by bacterial organisms. They can be isolated and used as such, but also, depending on the presence of basic or weak groups in the molecule, in the form of salts with pharmaceutically tolerable organic or inorganic acids or bases. Examples of suitable acids are hydrochloric, hydrobromic, sulfuric, phosphoric, acetic, tartaric, citric and fumaric. Examples of suitable bases are sodium hydroxide, potassium hydroxide, calcium hydroxide, aluminum hydroxide, ammonium hydroxide, nontoxic amines such as tri-alkylamines, recover triethylamine, procaine, dibenzylamine, N-benzyl - β-phenylethylamine, 1-ephenamine, Ν , dehydro-abietylamine, N, N ^-bis-dehydroabiethylethylenediamine, N- (lower) alkylpiperidine (e.g., N-ethylpiperidine) and other bases used to prepare salts with penicillins, 3 145899

Sidekæden i penicillinstrukturen med den almene formel Ϊ kan indeholde et asymmetrisk center. Afhængigt af konfigurationen omkring dette center vil forbindelserne forekomme i forskellige diastereo-isomere former, som alle er biologisk aktive. Ligeledes kan ester-grupperne indeholde asymmetriske carbonatomer, f.eks. når R = CH^, hvilket giver anledning til forskellige diastereoisome-re former, som også alle er biologisk aktive. Det vil forstås, at opfindelsen omfatter såvel de rene diastereoisomere som blandinger heraf.The side chain of the penicillin structure of the general formula Ϊ may contain an asymmetric center. Depending on the configuration around this center, the compounds will occur in various diastereoisomeric forms, all of which are biologically active. Likewise, the ester groups may contain asymmetric carbon atoms, e.g. when R = CH 2, giving rise to various diastereoisomeric forms, all of which are also biologically active. It will be understood that the invention encompasses both the pure diastereoisomers and mixtures thereof.

Det er kendt, at penicillansyrederivater med den almene formel IIIt is known that penicillanic acid derivatives of the general formula II

R\ /CH3R 1 / CH 3

- CH = N - CH - CH II- CH = N - CH - CH II

/ II 1°¾/ II 1 ° ¾

CO-N-CH - COOHCO-N-CH - COOH

1 2 hvor R og R sammen betegner - n“, hvor n er et tal 3-7, har stærk antibakteriel aktivitet, især mod gramnegative organismer (hollandsk patentbeskrivelse nr. 7016435). Sådanne forbindelser absorberes imidlertid ikke godt, når de administreres ad oral vej, og det er nødvendigt at administrere dem ved injektion. Ved den foreliggende opfindelse tilvejebringes estere af forbindelserne med formel II, hvilke estere absorberes godt oralt og derefter hydrolyseres i legemet til opnåelse af blod- og organniveauer af forbindelserne med den almene formel II, som er tilstrækkelige til behandling af infektionssygdomme forårsaget af bakterier, som er følsomme over for penicillansyrer med den almene formel II. Til opnåelse af fuld antibakteriel virkning af penicillansyrerne med formlen II er det nødvendigt at vælge sådanne estergrupper, som hurtigt hydrolyseres in vivo. Det er et vigtigt aspekt ved den foreliggende opfindelse , at de ved fremgangsmåden ifølge opfindelsen fremstillede forbindelser indeholder sådanne estergrupper, som hurtigt hydrolyseres i legemet efter oral absorption.1 2 where R and R together represent - n ", where n is a number 3-7, has strong antibacterial activity, especially against gram-negative organisms (Dutch Patent Specification No. 7016435). However, such compounds are not well absorbed when administered by oral route and it is necessary to administer them by injection. The present invention provides esters of the compounds of formula II which are well absorbed orally and then hydrolyzed in the body to obtain blood and organ levels of the compounds of general formula II sufficient to treat infectious diseases caused by bacteria which are sensitive to penicillanic acids of general formula II. In order to achieve full antibacterial activity of the penicillanic acids of formula II, it is necessary to select such ester groups which are rapidly hydrolyzed in vivo. It is an important aspect of the present invention that the compounds prepared by the process of the invention contain such ester groups which are rapidly hydrolyzed in the body after oral absorption.

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Fra dansk patentansøgning nr. 5650/70 kendes forbindelser, som er nært beslægtede med de her omhandlede forbindelser, idet begge klasser af forbindelser indeholder et strukturelement: R1 \ N-CH=N- / R2 og idet de af dansk patentansøgning nr. 5650/70 omfattede forbindel- 3 3 ser endvidere gennem strukturelementet -CO-R , hvor R betegner eventuelt substitueret hydroxyl, indbefatter en række estere.From Danish Patent Application No. 5650/70, compounds which are closely related to the compounds of the present invention are known, both classes of compounds having a structural element: R1 \ N-CH = N- / R2 and those of Danish patent application No. 5650 / 70 also included compounds through the structural element -CO-R, wherein R represents optionally substituted hydroxyl, including a variety of esters.

Dansk patentansøgning nr. 5650/70 beskriver eller antyder imidlertid ikke forbindelser, som i denne esterdel indeholder den carbonat-struktur, der er karakteristisk for de ved fremgangsmåden ifølge opfindelsen fremstillede forbindelser. Til påvisning, henholdsvis sandsynliggørelse af de her omhandlede forbindelsers overlegne farmakologiske virkning i sammenligning med de fra dansk patentansøgning nr. 5650/70 kendte forbindelser er udført følgende forsøg.However, Danish Patent Application No. 5650/70 does not disclose or imply compounds which contain in this ester moiety the carbonate structure characteristic of the compounds prepared by the process of the invention. For the purpose of demonstrating and proving, respectively, the superior pharmacological action of the compounds of this invention in comparison with the compounds known from Danish Patent Application No. 5650/70, the following experiments have been carried out.

Der foretages sammenligning af de farmakologiske egenskaber hos den foretrukne her omhandlede forbindelse med formlen AComparison of the pharmacological properties of the preferred compound of formula A herein is made

H,C-CH>CH,. cH, C-CH> CH ,. c

2 ^ 2\ /S\ /CH2 ^ 2 \ / S \ / CH

N-CH=N-CH - CH C AN-CH = N-CH - CH C A

1 ά Δ CH, CH, 0 3 I 3 Η CO - N- CH-COO - CH-0-C-0-C2H51 ά Δ CH, CH, 0 3 I 3 Η CO - N- CH-COO - CH-0-C-0-C2H5

og hos den foretrukne blandt de i henhold til dansk patentansøgning nr. 5650/70 fremstillede forbindelser med formlen Band in the preferred of the compounds of formula B prepared according to Danish Patent Application No. 5650/70

H2C-CH2_CH2^ N-CH=N-CH - CH CxH2C-CH2_CH2 ^ N-CH = N-CH - CH Cx

H2C-CH2-CH2"" II I CH3 0 CH3 BH2C-CH2-CH2 "" II I CH3 0 CH3 B

CO _ N-CH- C00- CH 2” 0- C- C- CH 3 ch3CO _ N-CH- C00- CH 2 ”0- C- C- CH 3 ch3

ved standardtest. Der udføres endvidere standardtest med den her omhandlede forbindelse Dby standard test. Furthermore, standard tests are carried out with the compound D in question

5 145699 / \ /\/CT3 N-CH=N-CH - CH C.N-CH = N-CH - CH C.

W Vch3 d CH3 co - n-ch-cooAh-ocooch(ch3) 2 til sairanenligning med den her omhandlede forbindelse A og den fra den danske patentansøgning kendte forbindelse B.W Vch3 d CH3 co - n-ch-cooAh-ocooch (ch3) 2 for comparison with the present invention A and the compound B known from the Danish patent application.

Forbindelserne A og D fremstillet ved fremgangsmåden ifølge den foreliggende opfindelse hydrolyseres ligesom forbindelse B, der er kendt fra dansk patentansøgning nr. 5650/70, i legemet, hvorved fås forbindelsen med formlen CCompounds A and D prepared by the process of the present invention are hydrolyzed, like compound B, known from Danish Patent Application No. 5650/70, in the body to give the compound of formula C

H„C-CH,-CHoV / S CH, 2 2 2 \ / \ X 3 N-CH=N-CH - CH C, / \ p H2C-CH2-CH2 x ch3 cH + C-CH, -CHoV / S CH, 2 2 2 \ / \ X 3 N-CH = N-CH - CH C, / \ p H2C-CH2-CH2 x ch3 c

CO - N-CH-COOHCO - N-CH-COOH

Forbindelse C er kendt som mecillinam, og den resorberes dårligt fra mavetarmsystemet. Det er formålet med forbindelserne A, B henholdsvis D at gøre forbindelsen C (mecillinam) terapeutisk tilgængelig ved oral absorption.Compound C is known as mecillinam and it is poorly resorbed from the gastrointestinal system. The purpose of compounds A, B and D, respectively, is to make compound C (mecillinam) therapeutically available by oral absorption.

Der anvendes følgende test til sammenligning af egenskaberne af de ovenfor beskrevne forbindelser: 1. Test på hund med hensyn til resorption og relativ biotilgængelighed af forbindelserne efter oral administration.The following tests are used to compare the properties of the compounds described above: 1. Test on dogs for resorption and relative bioavailability of the compounds after oral administration.

2. Test på mus med hensyn til resorption og relativ biotilgængelighed af forbindelserne efter oral administration.2. Tests on mice for resorption and relative bioavailability of the compounds after oral administration.

3. Opløselighed af forbindelserne A og B i vand, i phosphatpuf fer og i syntetisk mavesaft.3. Solubility of compounds A and B in water, in phosphate buffer and in synthetic gastric juice.

6 145699 1. Test på hund med hensyn til resorption og relativ biotilgængelighed efter oral administration.6 145699 1. Test on dog for resorption and relative bioavailability after oral administration.

De to estere af mecillinam A og B administreres oralt i et overkrydsningsforsøg med B,eagle-hunde. Der anvendes fire Beagler, to hanner og to hunner med en legemsvægt på 11 - 18 kg.The two esters of mecillinam A and B are administered orally in a crossover experiment with B, eagle dogs. Four Beagles, two males and two females with a body weight of 11 - 18 kg are used.

Teststoffer.Test substances.

Forbindelse A i en renhed på 93%, forbindelse B i en renhed på 89%, forbindelse C (mecillinam) anvendes som referenceforbindelse i forsøget.Compound A at a purity of 93%, Compound B at a purity of 89%, Compound C (mecillinam) is used as the reference compound in the experiment.

Begge estere anvendes som hydrochlorid.Both esters are used as hydrochloride.

Forsøgsudførelse.Experimental Design.

Forsøget udføres som 2-vejs overkrydsningsforsøg på 2 dage med et interval på 4 dage ifølge nedenstående skema:The test is carried out as a 2-day cross-over experiment in 2 days with a 4-day interval according to the following schedule:

Hund nr. Køn Legemsvægt Testdato 77-04-14 Testdato 77-04-18 i kgDog No. Gender Body weight Test date 77-04-14 Test date 77-04-18 in kg

49 han 12 Forbindelse A Forbindelse B49 male 12 Compound A Compound B

50 han 18 Forbindelse B Forbindelse A50 male 18 Compound B Compound A

51 hun 12 Forbindelse A Forbindelse B51 female 12 Compound A Compound B

52 hun 11 Forbindelse B Forbindelse A52 female 11 Compound B Compound A

Esterne indgives i hårde gelatinekapsler i en mængde, der er ækvivalent med 20 mg mecillinam pr. kg legemsvægt. Hundene lades faste natten over, men har fri adgang til vand.The esters are administered in hard gelatin capsules in an amount equivalent to 20 mg mecillinam per ml. kg body weight. The dogs are allowed to stay overnight, but have free access to water.

Der udtages blodprøver fra vena jugularis før og 15 minutter, 30 minutter, 1 time, 1 time og 30 minutter, 2 timer, 3 timer, 4 timer og 6 timer efter administrationen. Prøverne centrifugeres, og serum fraskilles.Blood samples are taken from the vena jugularis before and 15 minutes, 30 minutes, 1 hour, 1 hour and 30 minutes, 2 hours, 3 hours, 4 hours and 6 hours after administration. The samples are centrifuged and serum is separated.

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Analyse:Analysis:

Prøven analyseres med hensyn til indholdet af mecillinam ved anvendelse af cylinderplademetoden med Escherichia coli Leo HA 2 som testorganisme.The sample is analyzed for the content of mecillinam using the cylinder plate method with Escherichia coli Leo HA 2 as the test organism.

Testresultat:Test Result:

Middelserumniveauerne af mecillinam (forbindelse C) angives i tabel I sammen med de enkelte middelværdier på topniveauerne. Arealet under serumkoncentrationskurven (AUC), som anvendes som en indikation på den relative biotilgængelighed i testforbindelserne, angives i tabel II. Arealet beregnes ved de overlappende parabolers metode.The mean serum levels of mecillinam (compound C) are given in Table I, together with the individual mean levels at the peak levels. The area under the serum concentration curve (AUC) used as an indication of the relative bioavailability of the test compounds is given in Table II. The area is calculated by the method of the overlapping parabolas.

8 145599 in8 145599 in

10 CO10 CO

CM OCM O.

3 *· -3 * · -

Q) ’ O OQ) 'O O

6 .+1 +1 •H CM CO6. + 1 + 1 • H CM CO

•p © ©• p © ©

+, «I+, «I

o(0 CO © © fto (0 CO © © ft

tJI g IDtJI g ID

«.nj ό r- M 3 3 ^ 3-3 0) O >«.Nj ό r- M 3 3 ^ 3-3 0) O>

OH g +1 OOH g +1 O

13 Η -Η © +1 M -3 +1 CM in CJ1 0 - " 3 0) co co •H g pj 0013 Η -Η © +1 M -3 +1 CM in CJ1 0 - "3 0) co co • H g pj 00

Mg in CMMg in CM

Ό 0 3 - ro >1 Μ 0) H -Ό 0 3 - ro> 1 Μ 0) H -

3 g +1 O3 g + 1 o

,^+) -H CO +1, + + - -H CO + 1

Mil) -P H COMil) -P H CO

(DC -- > tji co co in Ο a) 3 r- o(DC -> tji co co in Ο a) 3 r- o

+) Ο co H+) Ο co H

(D X) 3 -- - H(D X) 3 - - H

,—, 0) rH i—l g -3-0 g +1 +1 \ tn -3 h co tn to λ: <u +i - - 3 \ M rH o \, -, 0) rH i — l g -3-0 g +1 +1 \ tn -3 h co tn to λ: <u + i - - 3 \ M rH o \

tJi O’ ι-H CM Η H CMtJi O 'ι-H CM Η H CM

O g (1) OO g (1) O

Ό 3 -Ό 3 -

rfj O C 0) Hrfj O C 0) H

CM -3 +> H CO 3 0) Λ +> C- ° fl) +1 3 ·· 3 3 - - 3CM -3 +> H CO 3 0) Λ +> C- ° fl) +1 3 ·· 3 3 - - 3

H O' 0 3 Η Η Ο MH O '0 3 Η Η Ο M

0) 3 *3 -3+1 +1 -3 -0) 3 * 3 -3 + 1 +1 -3 -

Η M -3 g H 00 +JOΗ M -3 g H 00 + JO

H 3 - - 3 HH 3 - - 3 H

H (DO) g O CM CO 3 111 On m © H +> fflH (DO) g O CM CO 3 111 On m © H +> ffl

Si 3 0 3 3 0 HO 3 /-. 3 CD 0)Si 3 0 3 3 0 HO 3 / -. 3 CD 0)

Eh Λ rH rH (D Ο MEh Λ rH rH (D Ο M

3 · H g -P CM O 3 rH3 · H g -P CM O 3 rH

O J) Η \ +) o r~ 0 CDO J) Η \ +) o r ~ 0 CD

iw Μ O O' 3 - - A! fl H <D 3 3 - Η H §3 UH 0) g \ -3 +1 +1 3-3iw Μ O O '3 - - A! fl H <D 3 3 - Η H §3 UH 0) g \ -3 +1 +1 3-3

id tji - g cd tn 3 Aid tji - g cd tn 3 A

•3 UH - - CD 3• 3 UH - - CD 3

«>(0 Ο Ο r- MO«> (0 Ο Ο r- MO

Ο MH © rH MHΟ MH © rH MH

-3 (0 3 CD-3 (0 3 CD

+> Ό 3 3 +> -+> Ό 3 3 +> -

(03(1) Ο) Γ- CO MH(03 (1) Ο) Γ- CO MH

3 (0 > -P Η O <Dg +1¾ Η -P - - -r> \ M3 3 3 H co -SCn -3 (0 g 3 X! 3 3 -P 3 -3 +1+1 \ -3 M 3 g 0) o3 (0> -P Η O <Dg + 1¾ Η -P - - -r> \ M3 3 3 H co -SCn -3 (0 g 3 X! 3 3 -P 3 -3 + 1 + 1 \ -3 M 3 g 0) o

g 0) © CM H CMg 0) © CM H CM

03+1 CQ O - - H -03 + 1 CQ O - - H -

(0 co CO O* 0) H(0 co CO O * 0) H

•3 3 H 03 3 Ό +1 (0 3 0 -3 3 « -P > ^ Ο > +> -3 - H 3 co ο* Ό co• 3 3 H 03 3 Ό +1 (0 3 0 -3 3 «-P> ^ Ο> +> -3 - H 3 co ο * Ό co

3 0) 3^ ["· 3 H3 0) 3 ^ ["· 3 H

Q) id -3 - - - -3 +> <d g ο o < 13-3 -w v 0) a) g m tS Q)Q) id -3 - - - -3 +> <d g ο o <13-3 -w v 0) a) g m tS Q)

Η MΗ M

3 - l 3 H3 - l 3 H

a) a) i o a) 3 T3 a) H MH Ό 3 3 M a) 3 0) 3 H f^rdCQX 3-3 > .3 CD +)3 -3 Ο) Si •3 0) 'd 0) 0) -3 Q) 3 -3 3a) a) ioa) 3 T3 a) H MH Ό 3 3 M a) 3 0) 3 H f ^ rdCQX 3-3> .3 CD +) 3 -3 Ο) Si • 3 0) 'd 0) 0 ) -3 Q) 3 -3 3

3H 3 MHiH 01 fti "dO3H 3 MHiH 01 fti „dO

g O’ -3 Η H ft *3 0) 3ftg O '-3 Η H ft * 3 0) 3ft

33Λ Q)-3 0 d)-P iB33Λ Q) -3 0 d) -P iB

30)3 Π3+) 03 >·· CDPQO 3 M 0) 3 +> H·30) 3 Π3 +) 03> ·· CDPQO 3 M 0) 3 +> H ·

M UH -3 g O’ -3 Ό Q)HM UH -3 g O '-3 Ό Q) H

*d 0) +> .0 0) H Λ 3 03·* d 0) +> .0 0) H Λ 3 03 ·

03 M 3 3 T9. 3 3 0) Ό CQ03 M 3 3 T9. 3 3 0) Ό CQ

H-3 0) Ο Ή 4H 0) 0,¾ -3 com Eh ft -- -3 M ft ~ S +1 9 145639H-3 0) Ο Ή 4H 0) 0, ¾ -3 com Eh ft - -3 M ft ~ S +1 9 145639

Tabel II.Table II.

Biotilgængelighed målt som arealet under serumkoncentrationskurven (AUC).Bioavailability measured as the area under the serum concentration curve (AUC).

Hund nr. AUC (timer x^ug x ml ^Dog No. AUC (hours x ^ ug x ml ^

Forbindelse A Forbindelse BCompound A Compound B

49 23,8 28,6 50 43,7 32,6 51 33,1 31,7 52 29,3 23,449 23.8 28.6 50 43.7 32.6 51 33.1 31.7 52 29.3 23.4

Middelværdi 32,5 '29,1 - S.D. 8,40 4,15 - S.E. 4,20 2,07Mean 32.5 '29, 1 - S.D. 8.40 4.15 - S.E. 4.20 2.07

Det fremgår af tabel I, at der kun er ubetydelige forskelle i de tidligere og sene middelværdier for serumniveauerne af mecillinam efter oral administration af forbindelse A og forbindelse B. 60 minutter efter administrationen giver forbindelse A imidlertid et markant højere niveau end forbindelse B (10,6 - 1,02 henholdsvis 7,5 - 1,70/Ug/ml), og disse højere niveauer er endnu tydelige ' -f J- 3 timer efter administrationen (6,18 - 1,58 henholdsvis 5,3 - 0,32 jyq/ml). Forbindelse A giver en middelværdi på de højeste serumniveauer på 12,1 - l,71yug/ml, der optræder 90 minutter efter administrationen, medens forbindelse B giver en topværdi på 10,3 - 1,10 ^ug/ml 2 timer efter administrationen. Dette senere niveau er imidlertid stadig lavere end det, der opnås med forbindelse A efter 2 timer (11,1 - 1,87/Ug/ml). Beregning af middelværdi for topniveauerne i serum uafhængigt af den tid, ved hvilken de indtræffer i de forskellige dyr, giver en værdi på 13,4 - l,20yug/ml for forbindelse A, hvil 10 145699 ket er ca. 20% højere end den, der opnås med den kendte forbin-delse B (10,9 - l,Q2^ug/ml). Statistisk analyse viser, at denne differens er signifikant ved p < 0,11.However, Table I shows that there are only negligible differences in the early and late mean levels of serum levels of mecillinam after oral administration of compound A and compound B. However, 60 minutes after administration, compound A provides a significantly higher level than compound B (10, 6 - 1.02 and 7.5 - 1.70 / ug / ml, respectively), and these higher levels are even more pronounced -f J- 3 hours after administration (6.18 - 1.58 and 5.3-0, respectively). 32 µg / ml). Compound A gives a mean value of the highest serum levels of 12.1-1.7 µg / ml occurring 90 minutes after administration, while Compound B gives a peak value of 10.3 - 1.10 µg / ml 2 hours after administration. However, this latter level is still lower than that achieved with Compound A after 2 hours (11.1 - 1.87 / ug / ml). Calculation of the mean serum peak levels, independent of the time at which they occur in the various animals, gives a value of 13.4-1.20 µg / ml for Compound A, which is about 10%. 20% higher than that obtained with the known compound B (10.9-1.1 Q2 µg / ml). Statistical analysis shows that this difference is significant at p <0.11.

Den relative biotilgængelighed for forbindelse A er højere end for forbindelse B i tre af fire dyr, der anvendes til forsøget.The relative bioavailability of compound A is higher than that of compound B in three of four animals used for the experiment.

Den middelværdi, der opnås for de relative biotilgængeligheder for den her omhandlede forbindelse A, er ca. 10% højere end de, der opnås med den kendte forbindelse B, eller 32,5 i sammenligning med 29,1 timer Xyiig/ml-^.The mean value obtained for the relative bioavailability of the present compound A is approx. 10% higher than those obtained with the known compound B, or 32.5 as compared to 29.1 hours Xy / µg / ml.

2. Test på mus med hensyn til resorption og relativ tilgængelighed efter oral administration.2. Test on mice for resorption and relative availability after oral administration.

2.1 Sammenligning af forbindelse A og forbindelse B.2.1 Comparison of Compound A and Compound B.

Forbindelse A (fremstillet ifølge opfindelsen) anvendes som hydrochlo-rid i renhed 97,4%, forbindelse B (kendt teknik) anvendes som fri base, smeltepunkt 117,5 - 119,5°C, og forbindelse C, mecillinam, anvendes som reference.Compound A (prepared according to the invention) is used as a hydrochloride in purity 97.4%, compound B (known in the art) is used as a free base, m.p. 117.5 - 119.5 ° C, and compound C, mecillinam, is used as reference. .

Hvide SPF-hunmus med en vægt på 20 - 22 g indgives 4 mg (ca. 200 mg/kg legemsvægt) af en af forbindelserne A, B eller C oralt som opløsning i 0,2 ml vand beregnet som amidinopenicillansyre (forbindelce C). Der udtages blodprøver 30 minutter, 60 minutter, 90 minutter, 2 timer og 3 timer efter administrationen. Fem mus anæstetiseres med chloroform ved hvert prøvetidspunkt. Blod udtages ved hjertepunktur og sammenblandes. Nogle minutter før denne fremgangsmåde hepariniseres musen med 0,1 ml heparin (1000 IE) intraperi-tonealt. Plasmaet skilles fra og testes for indholdet af mecillinam ved cylinderplademetoden og med Escherichia coli 1008 som testorganisme.White SPF female mice weighing 20 to 22 g are administered 4 mg (about 200 mg / kg body weight) of one of the compounds A, B or C orally as a solution in 0.2 ml of water calculated as amidinopenicillanic acid (compound C). Blood samples are taken 30 minutes, 60 minutes, 90 minutes, 2 hours and 3 hours after administration. Five mice are anesthetized with chloroform at each sample time. Blood is taken at heart puncture and mixed. A few minutes before this procedure, the mouse is heparinized with 0.1 ml of heparin (1000 IU) intraperitoneally. The plasma is separated and tested for the content of mecillinam by the cylinder plate method and with Escherichia coli 1008 as the test organism.

Testresultat.Test results.

Serumniveauerne af amidinopenicillansyre (forbindelse C) efter oral administration af forbindelserne A, B og C i ækvimolære doser på ll,45^,umol pr. dyr er angivet i tabel III. Arealet under serumkoncentrationskurven (AUC) er angivet i tabel IV.Serum levels of amidinopenicillanic acid (Compound C) after oral administration of Compounds A, B and C in equimolar doses of 11, 45 µmol per day. animals are listed in Table III. The area under the serum concentration curve (AUC) is given in Table IV.

1X 1456931X 145693

Tabel IIITable III

Serumniveauer af mecillinam efter oral administration af ækvi-molære mængder af forbindelserne A, B og C.Serum levels of mecillinam after oral administration of equimolar amounts of compounds A, B and C.

Testforbindelse Serumniveau (^ug/ml) 30 minutter 60 minutter 90 minutter 2 timer 3 timerTest compound Serum level (µg / ml) 30 minutes 60 minutes 90 minutes 2 hours 3 hours

Forbindelse A (fremstillet i-følge opfindelsen) (hydro- chlorid) 42 31 14 10 3,8Compound A (prepared according to the invention) (hydrochloride) 42 31 14 10 3.8

Forbindelse B (kendt teknik) (fri base) 27 16 22 13 5,6Compound B (Prior Art) (Free Base) 27 16 22 13 5.6

Forbindelse CCompound C

(reference) 4,0 3,0 2,2 1,0 0,5(reference) 4.0 3.0 2.2 1.0 0.5

Tabel IVTable IV

Relativ biotilgængelighed målt som arealet under serumkoncentrationskurven (AUC).Relative bioavailability measured as the area under the serum concentration curve (AUC).

Testforbindelse AUC/timer x^ug x ml ^Test compound AUC / hr x ^ ug x ml ^

Forbindelse A (.fremstillet ifølge opfindelsen) 55,7Compound A (prepared according to the invention) 55.7

Forbindelse B (kendt teknik) 48,1Compound B (Prior Art) 48.1

Det fremgår af tabel III, at forbindelse A giver højere initiale serumniveauer end forbindelse B. Efter 90 minutter og 2 og 3 timer efter administrationen giver den kendte forbindelse B højere niveauer end forbindelse A. Den relative biotilgængelighed af for- 12 145699 bindelse A fremstillet ifølge opfindelsen er imidlertid bedre end den relative biotilgængelighed for forbindelse B, 55,7 i sammenligning med 46,1 timer x^ug x ml"-1, 2.2 Sammenligning af forbindelse A, B og D.It is evident from Table III that Compound A gives higher initial serum levels than Compound B. After 90 minutes and 2 and 3 hours after administration, the known Compound B gives higher levels than Compound A. The relative bioavailability of Compound A prepared according to however, the invention is better than the relative bioavailability of Compound B, 55.7 as compared to 46.1 hours x µg x ml "-1, 2.2 Comparison of Compounds A, B and D.

Der anvendes den i henhold til den foreliggende opfindelse fremstillede forbindelse A i en renhed på 93%, den kendte forbindelse B i en renhed på 89%, og den her omhandlede forbindelse D i en renhed på 95%. Alle forbindelser anvendes som hydrochlorider. Der anvendes forbindelse C, mecillinam, som reference ved forsøget.The compound A prepared according to the present invention is used in a purity of 93%, the known compound B in a purity of 89% and the present compound D in a purity of 95%. All compounds are used as hydrochlorides. Compound C, mecillinam, is used as reference in the experiment.

Forsøget udføres som beskrevet ovenfor i afsnit 2.1.The experiment is performed as described above in section 2.1.

Testresultat.Test results.

Serumniveauerne af mecillinam (forbindelse C) efter oral administration af teststofferne i ækvimolære doser (ll,45yUmol pr. dyr) angives i tabel V. Den relative biotilgængelighed beregnet som arealet under serumkoncentrationskurven (AUC) angives i tabel VI.The serum levels of mecillinam (compound C) after oral administration of the test substances at equimolar doses (II, 45 µmol per animal) are given in Table V. The relative bioavailability calculated as the area under the serum concentration curve (AUC) is given in Table VI.

Tabel VTable V

Serumniveauer for mecillinam efter oral administration af ækvimolære mængder af forbindelse A, B og D.Serum levels of mecillinam after oral administration of equimolar amounts of compounds A, B and D.

Testforbindelse Serumniveauer (^ug/ml) 15 min'. 30 min. 60 min. 90 min. 2 timer 3 timer 4 timerTest compound Serum levels (µg / ml) 15 min. 30 min. 60 min. 90 min. 2 hours 3 hours 4 hours

Forbindelse A (fremstillet ifølge,°pfin- 136 98 31 10 3,5 9,0 *> 0,97Compound A (prepared according to ° pfin 136.0 31 10 3.5 9.0 *> 0.97

Forbindelse BCompound B

(kendt teknik) 83 36 20 17 11 9,1 6,7(prior art) 83 36 20 17 11 9.1 6.7

Forbindelse D (fremstillet ifølge opfin- 130 102 35 17 6,5 2,8 1,9 delsen) _____Compound D (prepared according to the invention)

Udeladt ved beregningen af AUC.Omitted when calculating AUC.

x) 13 145699x) 13 145699

Tabel VITable VI

Relativ biotilgængelighed· målt som arealet under serumkoncentrationskurven (AUC).Relative bioavailability · measured as the area under the serum concentration curve (AUC).

Testforbindelse AUC, timer x^ug x ml 1Test compound AUC, hours x µg x ml 1

Forbindelse A (fremstillet ifølge opfindelsen) 98Compound A (made according to the invention) 98

Forbindelse B (kendt teknik) 74Compound B (Prior Art) 74

Forbindelse D (fremstillet ifølge opfindelsen) 107Compound D (made according to the invention) 107

Det fremgår af tabel V, at de her omhandlede forbindelser (A og D) har højere topniveauer end forbindelse B efter 15 minutter.Table V shows that the compounds of this invention (A and D) have higher peak levels than compound B after 15 minutes.

30'minutter efter administrationen giver de her omhandlede forbindelser mecillinamniveauer, som er 2,7 - 2,8 gange højere end de, der opnås med forbindelse B. 1,5 timer efter administrationen er det niveau, der opnås med forbindelse B, noget højere end det, der opnås med forbindelse A, og det samme, der opnås med forbindelse D. På senere tidspunkter er de niveauer, der opnås med forbindelse B, højere end de, som opnås med forbindelserne A og D. Den relative biotilgængelighed for forbindelserne er imidlertid 32 og 45% højere for forbindelse A henholdsvis forbindelse D end den, der opnås med forbindelse B.30 minutes after administration, the compounds of this invention give mexillin levels which are 2.7 - 2.8 times higher than those obtained with compound B. 1.5 hours after administration, the level achieved with compound B is somewhat higher. than that obtained with compound A and the same obtained with compound D. At later times, the levels obtained with compound B are higher than those obtained with compounds A and D. The relative bioavailability of the compounds is however, 32 and 45% higher for compound A and compound D, respectively, than that obtained with compound B.

Opløselighed af teststofferne.Solubility of the test substances.

Tabel VIITable VII

Opløseligheden af teststofferne som hydrochlorider i vand, phos-phatpuffer ved pH-værdi 6,5 og syntetisk mavesaft (pH-værdi 1,2).The solubility of the test substances such as hydrochlorides in water, phosphate buffer at pH 6.5 and synthetic gastric juice (pH 1.2).

14 14569914 145699

Testforbindelse Opløselighed (g/100 ml)Test compound Solubility (g / 100 ml)

Vand Phosphatpuffer Syntetisk mavesaft pH-værdi 6,5 pJ.værdl lf2Water Phosphate Buffer Synthetic gastric juice pH 6.5 p. Value lf2

Forbindelse A (fremstillet 7,3Compound A (prepared 7.3

Forbindelse BCompound B

(kendt teknik) 3,4 1,2 15(prior art) 3.4 1.2 15

Opløseligheden for den i henhold til den foreliggende opfindelse fremstillede forbindelse A er signifikant højere end opløseligheden for den kendte forbindelse B.The solubility of the compound A prepared according to the present invention is significantly higher than the solubility of the known compound B.

Det er af kritisk betydning for behandlingen af bakterieinfektioner med oralt administreret antibiotika såsom 6-aminopenicillansyre-derivater at opnå en resorption, som er så fuldstændig og hurtig som mulig. Jo mere fuldstændig resorption, der opnås, desto bedre vil stoffet udnyttes og således give højere mængder af antibiotisk aktivt stof ved infektionsstedet. Desuden reducerer en mere fuldstændig resorption risikoen for uønsket samvirken mellem stofferne og tarmens mikroflora, hvilket kan give anledning til bivirkninger hos patienten eller fremme tilvækst og spredning af sådanne bakteriestammer i mikrofloraen, der er resistente over for stoffet. En hurtigere resorption giver anledning til en hurtigere indsætning af den antibakterielle virkning på infektionsstedet, men det giver også anledning til højere og tidligere topniveauer i serum, hvilket på sin side bevirker en bedre og hurtigere penetration af antibiotiket i kroppens væv og organer. Dette svarer til, at intravenøs administration, som giver en særdeles hurtig og komplet resorption, anvendes i tilfælde af alvorlige infektioner, hvor en så effektiv behandling som mulig kræves.It is critical to treat bacterial infections with orally administered antibiotics such as 6-aminopenicillanic acid derivatives to obtain a resorption that is as complete and rapid as possible. The more complete resorption achieved, the better the drug will be utilized, thus providing higher levels of antibiotic active substance at the site of infection. In addition, a more complete resorption reduces the risk of undesirable interaction between the substances and the gut microflora, which can cause side effects in the patient or promote the growth and spread of such bacterial strains in the microflora resistant to the drug. A faster resorption gives rise to a faster insertion of the antibacterial effect at the site of infection, but it also gives rise to higher and earlier serum levels, which in turn causes better and faster penetration of the antibiotic into the body's tissues and organs. This is similar to the fact that intravenous administration, which provides a very rapid and complete resorption, is used in cases of severe infections where as effective treatment as possible is required.

15 145699145699

Det fremgår således af de ovenfor beskrevne forsøg, at de her omhandlede forbindelser er lettere opløselige i maven og derfor hurtigere resorberes i organismen, hvilket giver anledning til en ønskelig hurtig opnåelse af et højt niveau for forbindelsen C (mecillinam), end den kendte forbindelse B, idet der fås højere og tidligere indtrædende topniveauer for den aktive forbindelse C ved administration af de her omhandlede forbindelser.Thus, it is apparent from the experiments described above that the compounds of this invention are more readily soluble in the stomach and therefore more rapidly resorbed in the organism, giving rise to a desirable rapid attainment of a high level of compound C (mecillinam) than the known compound B , with higher and earlier onset peak levels of active compound C being obtained by administering the compounds of this invention.

På grund af de særlig gode farmakologiske egenskaber for forbindelse A går et foretrukket aspekt af den foreliggende opfindelse ud på, at denne forbindelse fremstilles.Due to the particularly good pharmacological properties of compound A, a preferred aspect of the present invention is that this compound is prepared.

De omhandlede forbindelser med den almene formel I tåles godt, de medfører en kun ringe bivirkningsfrekvens, og de kan let anvendes i farmaceutiske præparater, enten som sådanne eller i form af deres salte, ligesom de let kan blandes med faste bærere eller adjuvanser eller begge dele. I sådanne præparater kan forholdet mellem det terapeutisk aktive stof og bærerne og adjuvanserne variere mellem 1% og 95%· Præparaterne kan være behandlede til dannelse af f.eks. tabletter, piller eller dragéer, eller de kan leveres i medicinske beholdere, f.eks. kapsler, eller de kan, for miksturers vedkommende, fyldes på flasker. Der kan til fremstillingen anvendes farmaceutisk tolerable, organiske eller uorganiske, faste eller flydende bærestoffer, som er egnede til oral eller enteral administration eller til topisk applikation. Gelatine, lactose, stivelse, magnesiumstearat, talkum, vegetabilske og animalske fedtstoffer og olier, vegetabilsk gummi og polyalkylenglycol og andre kendte bærestoffer for farmaceutiske præparater er alle velegnede til fremstilling af præparater med de omhandlede forbindelser. Præparaterne kan desuden indeholde andre farmaceutisk aktive komponenter, som er godt administrerbare sammen med de omhandlede forbindelser ved behandling af infektionssygdomme. Som eksempler på sådanne egnede antibiotisk virksomme stoffer kan nævnes gentamycin og polymyxin.The present compounds of general formula I are well tolerated, they have a low frequency of adverse reactions, and can be readily used in pharmaceutical preparations, either as such or in the form of their salts, and readily admixed with solid carriers or adjuvants or both. . In such compositions, the ratio of the therapeutically active substance to the carriers and adjuvants may vary between 1% and 95%. The compositions may be treated to form e.g. tablets, pills or dragees, or they can be delivered in medical containers, e.g. capsules, or for bottles, they can be filled into bottles. Pharmaceutically tolerable, organic or inorganic, solid or liquid carriers suitable for oral or enteral administration or for topical application may be used in the preparation. Gelatin, lactose, starch, magnesium stearate, talc, vegetable and animal fats and oils, vegetable gum and polyalkylene glycol and other known carriers for pharmaceutical compositions are all well suited for the preparation of compositions of the subject compounds. The compositions may additionally contain other pharmaceutically active components which are well administrable with the compounds of the present invention in the treatment of infectious diseases. Examples of such suitable antibiotic active agents are gentamycin and polymyxin.

Ved behandlingen af bakteriéLle infektioner i mennesker administreres de her omhandlede forbindelser f.eks. i mængder svarende til 5 - 200 mg/kg/dag, fortrinsvis i mængder i området 10 - 100 mg/kg/dag i inddelte doser, f.eks. 2, 3 eller 4 gange dagligt. De administreres i dosisenheder, som f.eks. indeholder 175 > 350, 500 eller 1000 mg af forbindelserne.In the treatment of bacterial infections in humans, the compounds of this invention are administered e.g. in amounts corresponding to 5 - 200 mg / kg / day, preferably in amounts in the range of 10 - 100 mg / kg / day in divided doses, e.g. 2, 3 or 4 times daily. They are administered in dosage units, e.g. contains 175> 350, 500 or 1000 mg of the compounds.

16 U5S9916 U5S99

Eksempler på. foretrukne forbindelser fremstillet ifølge opfindel— sen er anført i nedenstående tabel A, hvor Me og Et henholdsvis betyder methyl og ethyl:Examples of. preferred compounds prepared according to the invention are listed in Table A below, where Me and Et are methyl and ethyl, respectively:

Tabel a Z'—N r5Table a Z'-N r5

(CH^n ^N- -i,oW(CH ^ n ^ N- -i, oW

pyrrolidyl-1 CH2OCOOEt hexahydro-lH-azepin-l-yl CH2OCOOEt hexahydro-1(2H)-azocinnyl CH2OCOOEt pyrrolidyl-1 CH(Me)OCOOEt hexahydro-lH-azepin-l-yl CH(Me)OCOOEt hexahydro-1(2H)-azocinnyl CH(Me)OCOOEtpyrrolidyl-1 CH2OCOOEt hexahydro-1H-azepin-1-yl CH2OCOOEt hexahydro-1 (2H) -azocinnyl CH2OCOOEt pyrrolidyl-1 CH (Me) OCOOEt hexahydro-1H-azepin-1-yl CH (Me) OCOOEt hexahydro ) -azocinnyl CH (Me) OCOOEt

De omhandlede forbindelser kan fremstilles ved i og for sig kendte metoder, f.eks. ved omsætning af et reaktivt derivat af et amid eller thioamid med den almene formel IIIThe compounds of this invention can be prepared by methods known per se, e.g. by reacting a reactive derivative of an amide or thioamide of the general formula III

(CH^l^N - CH = R6 III(CH2-1 ^ N - CH = R6 III

g hvor n er et tal 3 - 7r og R betegner O eller S, med en ester afg where n is a number 3 - 7r and R represents O or S, with an ester of

6-aminopenicillansyre med den almene formel IV6-aminopenicillanic acid of the general formula IV

s ,CH3s, CH3

H0IT - CH - CHH0IT - CH - CH

II l^f 4 IVII l ^ f 4 IV

CO — N-CH - COO-CH-O-COO-R1CO - N-CH - COO-CH-O-COO-R1

De reaktive derivater af amiderne eller thioamiderne med formlen III er syreamidhalogenider eller dialkylsulfatcomplexer af syreamid- acetaler og kan fremstilles efter i og for sig kendte metoder ved 5 hvor R og R har den ovenfor anførte betydning.The reactive derivatives of the amides or thioamides of formula III are acid amide halides or dialkyl sulfate complexes of acid amide acetals and can be prepared by methods known per se at 5 wherein R and R are as defined above.

17 145699 behandling af forbindelserne med formlen III med halogeneringsmidler, f.eks. phosgen, oxalyldichlorid, thionylchlorid eller thionylbro-mid, eller med et dialkylsulfat såsom dimethylsulfat. Omsætningerne med halogeneringsmidler foretages i inerte tørre organiske opløsningsmidler såsom diethylether, toluen, benzen, chloroform eller carbontetrachlorid. Halogeniderne fås sædvanligvis i form af hygroskopiske præcipitater.Treatment of the compounds of formula III with halogenating agents, e.g. phosgene, oxalyldichloride, thionyl chloride or thionyl bromide, or with a dialkyl sulfate such as dimethyl sulfate. The reactions with halogenating agents are carried out in inert dry organic solvents such as diethyl ether, toluene, benzene, chloroform or carbon tetrachloride. The halides are usually obtained in the form of hygroscopic precipitates.

Ved behandlingen af syreamid-dialkylsulfat-complexerne med stærke baser, f.eks. natriummethoxid, omdannes de til syreamidacetaler med den almene formel VIn the treatment of the acid amide-dialkyl sulfate complexes with strong bases, e.g. sodium methoxide, they are converted to acid amide acetals of the general formula V

(cCTk - ch(or7)2 V(cCTk - ch (or7) 2 V

v H / 7 hvor n har den ovenfor angivne betydning, og R betegner en lavere alkylgruppe, som stammer fra dialkylsulfat, som også kan omsættes med esterne af 6-aminopenicillansyre med formlen IV, hvorved der fås forbindelserne med formlen I.v H / 7 where n is as defined above and R represents a lower alkyl group derived from dialkyl sulfate which can also be reacted with the esters of 6-aminopenicillanic acid of formula IV to give the compounds of formula I.

Esterne af 6-aminopenicillansyre med den almene formel IV kan fremstilles ved behandling af 6-APA med forbindelserne med formlen R5 1 1 4The esters of 6-aminopenicillanic acid of general formula IV can be prepared by treating 6-APA with the compounds of formula R5 1 1 4

Y -CH-O-COO-RY -CH-O-COO-R

4 5 1 hvor R og R har den ovenfor anførte betydning, og Y betegner halogen eller en funktionel ækvivalent gruppe, som kan omsættes med en carboxygruppe under dannelse af en esterbinding, f.eks. en organisk sulfonsyrerest. Omsætningen udføres fortrinsvis i organiske opløsningsmidler såsom dimethylformamid, dimethylsulfoxid eller hexamethylphosphoramid.Wherein R and R are as defined above and Y represents halogen or a functional equivalent group which can be reacted with a carboxy group to form an ester bond, e.g. an organic sulfonic acid residue. The reaction is preferably carried out in organic solvents such as dimethylformamide, dimethylsulfoxide or hexamethylphosphoramide.

Alternativt behandles 6-acylaminopenicillansyrer med acylgrupper, som kan fjernes uden ødelæggelse af penicillinringsystemet, med forbindelserne med formlen 18 145699 R5 il 4Alternatively, 6-acylaminopenicillanic acids are treated with acyl groups which can be removed without destroying the penicillin ring system with the compounds of formula 18

Y -CH-O-COO-RY -CH-O-COO-R

4 5 1 hvor R , R og Y har den oveiifor anførte betydning, hvorved der fås estere af 6-acylaminopenicillansyrer, fra hvilke acylgrupperne derpå fjernes til dannelse af estere af 6-axninopenicillansyre med den almene formel III, En foretrukken fremgangsmåde består i, at man omsætter et salt, f,eks. natrium- eller kaliumsaltet af benzylpenicillin, med en forbindelse med formlen R5 il 4Wherein R, R and Y are as defined above to give esters of 6-acylaminopenicillanic acids, from which the acyl groups are then removed to form esters of 6-axininopenicillanic acid of the general formula III. one converts a salt, e.g. the sodium or potassium salt of benzylpenicillin, with a compound of formula R5 to 4

Y -CH-O-COO-RY -CH-O-COO-R

i et organisk opløsningsmiddel såsom dimethylfomamid, dimethyl-sulfoxid eller hexamethylphosphoramid eller i en blanding af et organisk opløsningsmiddel og vand, f.eks. vandigt acetone eller vandigt dioxan, til dannelse af den tilsvarende ester af benzylpenicillin. Phenylacetylsidekæden fjernes derpå efter den metode, som er beskrevet i hollandsk patentbeskrivelse nr. 6401421 eller sydafrikansk patentbeskrivelse nr. 67/2927, ved behandling med phosphorpentachlorid i nærværelse af en tertiær organisk base til dannelse af et iminochlorid, som omsættes med en alkohol såsom propanol til dannelse af den tilsvarende iminoether, som hydrolyseres ved tilsætning af vand til dannelse af esteren med formlen III. Alternativt kan phenylacetylsidekæden fjernes ved enzymatisk hydrolyse under anvendelse af Escherichia coli-acylase efter den fremgangsmåde, som er beskrevet i fransk patentskrift nr. 1576027.in an organic solvent such as dimethyl fomamide, dimethylsulfoxide or hexamethylphosphoramide or in a mixture of an organic solvent and water, e.g. aqueous acetone or aqueous dioxane to form the corresponding ester of benzylpenicillin. The phenylacetyl side chain is then removed by the method described in Dutch Patent Specification No. 6401421 or South African Patent Specification No. 67/2927, by treatment with phosphorus pentachloride in the presence of a tertiary organic base to form an imino chloride which is reacted with an alcohol such as propanol. forming the corresponding imino ether which is hydrolyzed by adding water to form the ester of formula III. Alternatively, the phenylacetyl side chain can be removed by enzymatic hydrolysis using Escherichia coli acylase following the procedure described in French Patent No. 1576027.

Ved en yderligere metode omsættes N-beskyttede 6-aminopenicillansyrer med en forbindelse med formlen R5 li 4In a further method, N-protected 6-aminopenicillanic acids are reacted with a compound of formula R5

Y -CH-O-COO-RY -CH-O-COO-R

til dannelse af den tilsvarende ester, fra hvilken beskyttelsesgrupperne fjernes til opnåelse af forbindelserne med den almene formel IV . Eksempler på anvendelige beskyttelsesgrupper er ben-zyloxycarbonylgruppen, som fjernes ved katalytisk hydrogenering, 19 145899 o-nitro-phenylsulfenylgruppen, som kan fjernes ved behandling med nucleofile midler ved sur pH-værdi (japansk patentbeskrivelse nr.to form the corresponding ester from which the protecting groups are removed to give the compounds of general formula IV. Examples of useful protecting groups are the benzyloxycarbonyl group which is removed by catalytic hydrogenation, the o-nitrophenylsulfenyl group which can be removed by treatment with nucleophilic agents at acidic pH (Japanese Patent Specification no.

505176), og tritylgruppen, som kan fjernes ved mild sur hydrolyse.505176), and the trityl group, which can be removed by mild acidic hydrolysis.

De omhandlede forbindelser kan fremstilles efter andre fremgangsmåder. β-Formamidopenicillansyre kan omdannes til estere med den almene formel VIThe compounds of this invention can be prepared by other methods. β-Formamidopenicillanic acid can be converted into esters of general formula VI

/CH/ CH

HCONHCH - CH C D R VIHCONHCH - CH C D R VI

I I CIi3 f CO-N- CH - COO-CE-O-COO-R4 4 5 hvor R og R har den ovenfor anførte betydning, ved omsætning med en forbindelse med formlen R5 1 I 4Wherein R and R have the meaning given above, by reacting with a compound of formula R5 1 I 4

Y -CH-O-COO-RY -CH-O-COO-R

under de ovenfor omtalte betingelser. Behandling af en forbindelse med formlen VI med en 1,1-dihalogendimethylether, f.eks. 1,1-di-chlordimethylether, i nærværelse af en tertiær organisk base, fører til dannelsen af et reaktivt derivat, som reagerer med en amin med den almene formel VIIunder the conditions mentioned above. Treatment of a compound of formula VI with a 1,1-dihalogen dimethyl ether, e.g. 1,1-Dichlorodimethyl ether, in the presence of a tertiary organic base, leads to the formation of a reactive derivative which reacts with an amine of the general formula VII

(CH2^n ^NH Vil hvor n har den ovenfor anførte betydning, hvorved der fås forbindelsen med formlen I.(CH2 ^ n ^ NH Will where n has the meaning given above to give the compound of formula I.

En yderligere fremgangsmåde består i omsætning af forbindelser med formlen II, hensigtsmæssigt i form af et salt, f.eks. en natrium-, kalium-, calcium- eller triethylammoniumsalt, med en forbindelse med formlen c R5 ] I 4A further process consists in reacting compounds of formula II, conveniently in the form of a salt, e.g. a sodium, potassium, calcium or triethylammonium salt, having a compound of formula c R 5] 4

Y -CH-O-COO-RY -CH-O-COO-R

20 145699 hvorved der fås de omhandlede forbindelser med den almene formel I. Omsætningen udføres hensigtsmæssigt i organiske opløsningsmidler såsom dimethylformamid, dimethylsulfoxid eller hexamethyl-phosphoramid eller i en blanding af vand og organiske opløsningsmidler såsom vandigt dioxan.The reaction is conveniently carried out in organic solvents such as dimethylformamide, dimethylsulfoxide or hexamethylphosphoramide or in a mixture of water and organic solvents such as aqueous dioxane.

Ved nogle af fremgangsmåderne til fremstilling af de omhandlede forbindelser kan'udgangsmaterialet være i form af et salt, f.elcs. et natrium-, kalium-, calcium- eller triethylammoniumsalt.In some of the processes for preparing the subject compounds, the starting material may be in the form of a salt, e.g. a sodium, potassium, calcium or triethylammonium salt.

Fremgangsmåden ifølge den foreliggende opfindelse til fremstilling af forbindelser med den almene formel IThe process of the present invention for the preparation of compounds of general formula I

^S\^CH3S ^ \ ^ CH3

(ch9) n-ch=n-ch - CH C \ I(ch9) n-ch = n-ch - CH C \ I

' i ch3 r5 CO - N-CH -coo-(!:h-o-coo-r1 eller terapeutisk acceptable syreadditionssalte deraf, i hvilken for- 4 mel n er et tal 3-7; R betegner en alkylgruppe med 1-8 carbonatomer 5 og R betegner hydrogen eller methyl, er således ejendommelig ved, atin CH3 R5 CO - N-CH-COO - (!: ho-COO-R1 or therapeutically acceptable acid addition salts thereof, in which formula 4 is a number 3-7; R represents an alkyl group of 1-8 carbon atoms 5 and R represents hydrogen or methyl is thus characterized by the fact that

a) et reaktivt derivat af en forbindelse med den almene formel IIIa) a reactive derivative of a compound of general formula III

(CH2^n >SV-CH=R6 III(CH2 ^ n> SV-CH = R6 III

hvor n har den ovenfor anførte betydning, og R er -0- eller -S-, omsættes med en forbindelse med den almene formel IVwhere n is as defined above and R is -O- or -S- is reacted with a compound of the general formula IV

/"S\ /CH3/ "S \ / CH3

H-N-CH - CH Cv IVH-N-CH - CH Cv IV

I CH3 R5 I 4I CH3 R5 I 4

CO - N-CH-C00-CH-0-C00-RCO - N-CH-C00-CH-0-C00-R

5 hvor R og R har den ovenfor anførte betydning; eller 21 1456995 wherein R and R are as defined above; or 21,55699

b) en forbindelse med den almene formel Vb) a compound of general formula V

----\ 0 - R7---- \ 0 - R7

(CH0) _ N-CH^ V(CH0) _ N-CH2 V

v y \ 7v y \ 7

0 - R0 - R

hvor n har den ovenfor anførte betydning, og R7 betegner en alkyl-gruppe, omsættes med en forbindelse med den almene formel IVwhere n is as defined above and R 7 represents an alkyl group is reacted with a compound of general formula IV

^ S\ ^CH3 h2n - ch - CH iv CH3 R5 I 4^ S \ ^ CH3 h2n - ch - CH iv CH3 R5 I 4

CO - N-CH - COO-CH-O-COO-RCO - N-CH - COO-CH-O-COO-R

4 5 hvor R og R har den ovenfor anførte betydning; ellerWherein R and R are as defined above; or

c) et reaktivt derivat af en forbindelse med den almene formel VIc) a reactive derivative of a compound of general formula VI

/CH3 HCONHCH - CH Cv/ CH3 HCONHCH - CH Cv

XCH0 R5 VIXCH0 R5 VI

I 4I 4

CO - N-CH - COO-CH-O-COO-RCO - N-CH - COO-CH-O-COO-R

4 54 5

hvor R og R har den ovenfor anførte betydning, omsættes med en forbindelse med den almene formel VIIwherein R and R are as defined above are reacted with a compound of general formula VII

(cOh vi1 hvor n har den ovenfor anførte betydning; eller(cOh vi1 where n has the meaning given above; or

d) en forbindelse med den almene formel IId) a compound of general formula II

y'-'N ^s\ ^chy '-' N ^ s \ ^ ch

(CH2)n N-CH=N-CH - CH C(CH2) n N-CH = N-CH - CH C

\__y CH3 II\ __ y CH3 II

CO - N-CH-COOHCO - N-CH-COOH

22 145699 hvor n har den ovenfor anførte betydning, omsættes med en forbindelse med den almene formel R5 1 1 4Wherein n has the meaning given above, it is reacted with a compound of the general formula R5 1 1 4

Y -CH-O-COO-RY -CH-O-COO-R

4 5 i hvor R og R har den ovenfor anførte betydning, og Y betegner halogen eller en funktionel ækvivalent gruppe, hvorefter den dannede forbindelse om ønsket omdannes til et salt deraf.Wherein R and R are as defined above, and Y represents halogen or a functional equivalent group, after which the compound formed, if desired, is converted to a salt thereof.

Fremgangsmåden ifølge opfindelsen belyses nærmere ved nedenstående eksempleriThe process according to the invention is illustrated in more detail by the following example

Eksempel 1.Example 1.

1’-Ethoxycarbonyloxyethyl-6-(hexahydro-lH-azepin-l-yl)-methylen-aminopenicillanat (fremgangsmådevariant b) .1'-Ethoxycarbonyloxyethyl 6- (hexahydro-1H-azepin-1-yl) methylene aminopenicillanate (process variant b).

3,1 g 1-hexamethyleniminocarboxaldehyd-dimethylaceta1 i 50 ml chloroform sættes dråbevis ved en temperatur på -30°C i løbet af 15 minutter til en opløsning af 5 g l’-ethoxycarbonyloxyethyl-S-amino-penicillanat og 1,9 ml triethylamin i 150 ml chloroform. Derpå lades temperaturen stige til 0°C i løbet af 30 minutter, og blandingen omrøres ved 0°C i løbet af yderligere 60 minutter. 120 ml vand tilsættes, og blandingen omrøres i yderligere 10 minutter. Vandfasen fraskiiles, og den organiske fase vaskes med vand og inddampes. Remanensen (5 g) viser et kraftigt IR-absorptionsbånd ved 1775 cm-1 (β-lactamring).3.1 g of 1-hexamethyleneiminocarboxaldehyde-dimethylacetyl in 50 ml of chloroform are added dropwise at a temperature of -30 ° C over 15 minutes to a solution of 5 g of 1'-ethoxycarbonyloxyethyl-S-amino-penicillanate and 1.9 ml of triethylamine. in 150 ml of chloroform. The temperature is then allowed to rise to 0 ° C over 30 minutes and the mixture is stirred at 0 ° C for a further 60 minutes. 120 ml of water are added and the mixture is stirred for a further 10 minutes. The aqueous phase is separated and the organic phase is washed with water and evaporated. The residue (5 g) shows a strong IR absorption band at 1775 cm -1 (β-lactam ring).

Inkubation af produktet med humant serum ved 37°C vises at forårsage en hurtig dannelse af 6-(hexahydro-lH-azepin-l-yl)-methylenaminopeni-cillansyre.Incubation of the product with human serum at 37 ° C is shown to cause a rapid formation of 6- (hexahydro-1H-azepin-1-yl) -methylenaminopenia-cillanoic acid.

11-Ethoxycarbonyloxyethyl-6-aminopenicillanatet fremstilles som be skrevet i svensk patentansøgning nr. 12688/70.The 11-ethoxycarbonyloxyethyl-6-aminopenicillanate is prepared as described in Swedish Patent Application No. 12688/70.

23 14569923 145699

Eksempel 2.Example 2.

Ved at erstatte l'-ethoxycarbonyloxyethyl-e-aminopenicillanatet ved en fremgangsmåde som beskrevet i eksempel 1 med ethoxycarbo-nyloxymethyl-6-aminopenicillanat fås ethoxycarbonyloxymethyl-6--(hexahydro-lH-azepin-l-yl)-methylenaminopenicillanat. Denne forbindelse viser en kraftig β-lactam-absorption i IR-spektret ved 1775 cm ^ og hydrolyseres hurtigt af humant serum til dannelse af den tilsvarende penicillansyre.By replacing the 1'-ethoxycarbonyloxyethyl-e-aminopenicillanate by a procedure described in Example 1 with ethoxycarboxyloxymethyl-6-aminopenicillanate, ethoxycarbonyloxymethyl-6- (hexahydro-1H-azepin-1-yl) -methyleneamopenicillanate is obtained. This compound shows a strong β-lactam absorption in the IR spectrum at 1775 cm 2 and is rapidly hydrolyzed by human serum to form the corresponding penicillanic acid.

Eksempel 3.Example 3

1'-Ethoxycarbonyloxyethyl-6-(piperidyl-1)-methylenamino-penicillanat.1'-ethoxycarbonyloxyethyl 6- (1-piperidyl) -methyleneamino-penicillanate.

Til 1,7 g (0,008 mol) phosphorpentachlorid og 1,8 g (0,016 mol) kinolin i 50 ml tørt methylenchlorid sættes 3,2 g (0,007 mol) l'--ethoxycarbonyloxyethylbenzylpenicillanat under omrøring og afkøling til -40°C. Reaktionen gennemføres i en tør inert· gas (argon). Efter 1 time tilsættes dråbevis 2,24 g tørt methanol, og temperaturen indstilles til -30°C. Efter 1 time sættes dråbevis 15 ml saltopløsning til opløsningen, medens temepraturen lades stige til 0°C.To 1.7 g (0.008 mole) of phosphorus pentachloride and 1.8 g (0.016 mole) of quinoline in 50 ml of dry methylene chloride add 3.2 g (0.007 mole) of 1'-ethoxycarbonyloxyethylbenzylpenicillanate with stirring and cooling to -40 ° C. The reaction is carried out in a dry inert gas (argon). After 1 hour, 2.24 g of dry methanol are added dropwise and the temperature is adjusted to -30 ° C. After 1 hour, 15 ml of brine are added dropwise to the solution while allowing the temperature to rise to 0 ° C.

Efter 15 minutter tørres den organiske fase og inddampes i vakuum. Remanensen behandles med petroleumsether og tørres, hvorved fås 2,8 g (100%'s udbytte) af en krystallinsk masse, IR-absorption: 1790 cm (β-lactam) . Til en opløsning af 2,8 g (0,007 mol) af dette materiale og 0,84 ml (0,006 mol) triethylamin i 50 ml chloroform sættes dråbevis ved -50 - -60°C 1,1 g (0,006 mol) N-piperidylchloro-formiminiumchlorid i 20 ml chloroform. Blandingen holdes under en tør argonatmosfære i 1,5 timer, i hvilken periode temperaturen stiger til 0°C. Opløsningsmidlet afdampes i vakuum ved 40°C, og remanensen opslemmes i 100 ml tørt acetone og filtreres. Filtratet inddampes i vakuum, og den som remanens vundne olie behandles med petroleumsether til krystallisation, udbytte 1,2 g, IR-absorption: 1760 cm (β-lactam), NMR: singlet ved 480 Hz (methylenimino), multiplet ved 320 Hz (penicillinkernens 5- og 6-stilling).After 15 minutes, the organic phase is dried and evaporated in vacuo. The residue is treated with petroleum ether and dried to give 2.8 g (100% yield) of a crystalline mass, IR absorption: 1790 cm (β-lactam). To a solution of 2.8 g (0.007 mol) of this material and 0.84 ml (0.006 mol) of triethylamine in 50 ml of chloroform is added dropwise at -50 to -60 ° C 1.1 g (0.006 mol) of N-piperidyl chloro -formiminium chloride in 20 ml of chloroform. The mixture is kept under a dry argon atmosphere for 1.5 hours, during which time the temperature rises to 0 ° C. The solvent is evaporated in vacuo at 40 ° C and the residue is slurried in 100 ml of dry acetone and filtered. The filtrate is evaporated in vacuo and the oil recovered as residue is treated with petroleum ether for crystallization, yield 1.2 g, IR absorption: 1760 cm (β-lactam), NMR: singlet at 480 Hz (methylenimino), multiplied at 320 Hz ( the 5 and 6 positions of the penicillin core).

24 14569924 145699

Eksempel 4.Example 4

Ethoxycarbonyloxymethyl-6-(hexahydro-lH-azepin-l-yl)-methylen-aminopenicillanat.Ethoxycarbonyloxymethyl-6- (hexahydro-lH-azepin-l-yl) methylene-aminopenicillanate.

Den i overskriften nævnte forbindelse fremstilles som beskrevet i eksempel 3 ud fra ethoxycarbonyloxyethylbenzylpenicillanat og 1,15 g (0,006 mol) hexahydro-lH-azepin-l-yl-chloroformiminium-chlorid, hvorved fås en olie. IR-absorption: 1760 cm-1 (β-lactam).The title compound is prepared as described in Example 3 from ethoxycarbonyloxyethylbenzylpenicillanate and 1.15 g (0.006 mol) of hexahydro-1H-azepin-1-yl chloroformiminium chloride to give an oil. IR absorption: 1760 cm -1 (β-lactam).

Eksempel 5.Example 5

11-Ethoxycarbonyloxyethyl-6-(hexahydro-lH-azepin-l-yl)-methylenanino-penicillanat. (Fremgangsmådevariant d)).11-ethoxycarbonyloxyethyl 6- (hexahydro-lH-azepin-l-yl) penicillanate -methylenanino. (Method variant d)).

1,02 g (3 mmol) tetrabutylammoniumhydrogensulfat opløses i 8,92 ml (9 mmol) 1,008 M natriumhydroxidopløsning under afkøling med is og omrøring. Der tilsættes 0,94 g (3 mmol) 6-(hexahydro-lH-azepin-yl)--methylenaminopenicillansyrehydrochlorid opløst i 15 ml ethanolfrit chloroform. Blandingen omrøres kraftigt i 30 minutter under afkøling med is. Den organiske fase fraskilles og tørres over natriumsulfat i 30 minutter under omrøring og afkøling med is. Opløsningen inddampes i vakuum, og remanensen opløses i 10 ml tørt acetone. Der tilsættes under omrøring og afkøling med is i løbet af 5 minutter 0,55 g (3,6 mmol) α-chlordiethylcarbonat opløst i 5 ml tørt acetone. Blandingen opvarmes til 50°C og omrøres i 6 timer ved 50°C. Opløsningen inddampes i vakuum, og remanensen opløses i 20 ml ethylacetat. Opløsningen vaskes med 15 ml vand, 10 ml 5%'s natriumbicarbonat-opløsning og 10 ml mættet natriumchloridopløsning, behandles med kul, tørres med natriumsulfat under afkøling på is og omrøring og inddampes i vakuum. Remanensen behandles med petroleumsether, hvorved fås 0,43 g 1'-ethoxycarbonyloxyethyl-6-(hexahydro-lH-azepin--1-yl)-methylenaminopenicillanat i form af en viskøs olie. Produktet har følgende NMR-spektrum i chloroform ved 60 MHz: NMR-spektrum (CDC13): δ(ppm) = 1,33 (3H,t,JHH = 7 Hz), 1,5 - 1,9 (17H, m), 3,38 (4H, m), 4,26 (2H, q, JRR = 7 Hz), l,33(t)JHR = 7Hz (3H), 1,5 - 1,9(m) (17H), 3,38(m) (4H), 4,26(q)JHH = 7 Hz (2H),Dissolve 1.02 g (3 mmol) of tetrabutylammonium hydrogen sulfate in 8.92 ml (9 mmol) of 1.008 M sodium hydroxide solution under cooling with ice and stirring. Add 0.94 g (3 mmol) of 6- (hexahydro-1H-azepin-yl) -methyleneamopenicillanic acid hydrochloride dissolved in 15 ml of ethanol-free chloroform. The mixture is stirred vigorously for 30 minutes while cooling with ice. The organic phase is separated and dried over sodium sulfate for 30 minutes with stirring and cooling with ice. The solution is evaporated in vacuo and the residue is dissolved in 10 ml of dry acetone. 0.55 g (3.6 mmol) of α-chlorodiethylcarbonate dissolved in 5 ml of dry acetone are added with stirring and cooling with ice over 5 minutes. The mixture is heated to 50 ° C and stirred for 6 hours at 50 ° C. The solution is evaporated in vacuo and the residue is dissolved in 20 ml of ethyl acetate. The solution is washed with 15 ml of water, 10 ml of 5% sodium bicarbonate solution and 10 ml of saturated sodium chloride solution, treated with charcoal, dried with sodium sulfate under cooling on ice and stirred and evaporated in vacuo. The residue is treated with petroleum ether to give 0.43 g of 1'-ethoxycarbonyloxyethyl-6- (hexahydro-1H-azepin-1-yl) methyleneaminopenicillanate in the form of a viscous oil. The product has the following NMR spectrum in chloroform at 60 MHz: NMR spectrum (CDCl3): δ (ppm) = 1.33 (3H, t, JHH = 7 Hz), 1.5 - 1.9 (17H, m) , 3.38 (4H, m), 4.26 (2H, q, JRR = 7 Hz), 1.33 (t) JHR = 7Hz (3H), 1.5 - 1.9 (m) (17H) , 3.38 (m) (4H), 4.26 (q) JHH = 7 Hz (2H),

4,37(d) (IH), 5,07(d)JHH = Hz (IH) , 5,46(d)JRH = 4 Hz (IH) , 6,82(dq)JHH4.37 (d) (1H), 5.07 (d) JHH = Hz (1H), 5.46 (d) JRH = 4 Hz (1H), 6.82 (dq) JHH

= 5 Hz (IH) = 7,61 (s) (IH).= 5 Hz (1H) = 7.61 (s) (1H).

Eksempel 6.Example 6

25 14569925 145699

Fremstilling af 1'-ethoxycarbonyloxyethyl-6fi-(hexahydro-lH-azepin--1-yl)-methylenaminopenicillanathydrochlorid (fremgangsmådevariant c) .Preparation of 1'-ethoxycarbonyloxyethyl-6β- (hexahydro-1H-azepin-1-yl) -methyleneamopenicillanate hydrochloride (process variant c).

A. Fremstilling af.1'-ethoxycarbonyloxyethyl-6j3-aminopenicillanat som udgangsmateriale.A. Preparation of 1'-ethoxycarbonyloxyethyl-6β-aminopenicillanate as starting material.

En opløsning af 7,4 g (0/016 mol) 1'-ethoxycarbonyloxyethylbenzyl-penicillanat i 10 ml tørt ethanolfrit chloroform sættes til en omrørt blanding af 3,8 g (0,018 mol) phosphorpentachlorid og 4,8 g (0,037 mol) quinolin i 25 ml tørt ethanolfrit chloroform ved -20°C i tør nitrogengasatmosfære. Efter omrøring ved denne temperatur i 1 time afkøles opløsningen til -30°C, og der tilsættes 12 ml tørt propanol i løbet af 20 minutter. Temperaturen lades stige til -15°C i løbet af 30 minutter, hvorefter der tilsættes 25 ml kold saltopløsning under kraftig omrøring, medens temperaturen stiger til -5°C. Reaktionsblandingen udhældes i 50 ml iskold ligroin og omrøres i 15 minutter, hvorefter der tilsættes yderligere 55 ml iskold ligroin, og blandingen omrøres i yderligere 10 minutter. Blandingen lades derpå separere, og olie-vand-fasen opsamles og ekstraheres med 30 ml chloroform. Chloroformfasen tørres ved 0°C over natriumsulfat, og olie-vand-fasen opsamles og ekstraheres med 30 ml chloroform. Chloroformfasen vaskes 2 gange med hver gang 100 ml iskold mættet natriumbicarbonatopløsning og tørres over natriumsulfat, og opløsningsmidlet afdampes under reduceret tryk. I infrarød analyse (CDClg) ses β-lactam (1775 cm ^) og ester (1755 cm ^). I NMR-spektrum (CDCl^) ses ved 4,51 ppm IH, d, CgH, J = 4,0 Hz, og ved 5,44 ppm IH, d, C^H, J = 4,0 Hz.A solution of 7.4 g (0.016 mole) of 1'-ethoxycarbonyloxyethylbenzyl penicillanate in 10 ml of dry ethanol-free chloroform is added to a stirred mixture of 3.8 g (0.018 mole) of phosphorus pentachloride and 4.8 g (0.037 mole) of quinoline. in 25 ml of dry ethanol-free chloroform at -20 ° C in dry nitrogen gas atmosphere. After stirring at this temperature for 1 hour, the solution is cooled to -30 ° C and 12 ml of dry propanol is added over 20 minutes. The temperature is allowed to rise to -15 ° C over 30 minutes, after which 25 ml of cold saline solution is added with vigorous stirring while the temperature rises to -5 ° C. The reaction mixture is poured into 50 ml of ice cold ligroin and stirred for 15 minutes, then an additional 55 ml of ice cold ligroin is added and the mixture is stirred for a further 10 minutes. The mixture is then allowed to separate and the oil-water phase is collected and extracted with 30 ml of chloroform. The chloroform phase is dried at 0 ° C over sodium sulfate and the oil-water phase is collected and extracted with 30 ml of chloroform. The chloroform phase is washed twice with 100 ml of ice-cold saturated sodium bicarbonate solution each time and dried over sodium sulfate, and the solvent is evaporated under reduced pressure. In infrared analysis (CDCl 3) β-lactam (1775 cm 2) and ester (1755 cm 2) are seen. In the NMR spectrum (CDCl3) is seen at 4.51 ppm 1H, d, CgH, J = 4.0 Hz, and at 5.44 ppm 1H, d, C1 H, J = 4.0 Hz.

B. Fremstilling af 1'-ethoxycarbonyloxyethyl-δβ-(hexahydro-lH--azepin-l-yl)-methylenaminopenicillanathydrochlorid.B. Preparation of 1'-Ethoxycarbonyloxyethyl-δβ- (hexahydro-1H-azepin-1-yl) -methyleneamopenicillanate hydrochloride.

Til en omrørt isafkølet opløsning af 3,3 g (0,01 mol) l'-ethoxy--carbonyloxyethyl-68-aminopenicillanat i 35 ml tørt ethanolfrit chloroform sættes 2,8 ml (0,02 mol) triethylamin og derpå 1,1 g (0,01 mol) 1,1-dichlordimethylether. Opløsningen lades henstå ved stuetemperatur natten over og afkøles derpå i isbad, hvorefter tilsættes l,lml (0,01 mol) hexamethylenimin. Blandingen opbevares 26 .145699 i køleskabet natten over, hvorefter opløsningsmidlet afdampes i vakuum. Remanensen fordeles mellem ethylacetat og vand. Efter tørring af ethylacetatfasen afdampes opløsningsmidlet, hvorved fås 3,8 g af en olieremanens, der indeholder ca. 80% af det ønskede produkt, ifølge dets NMR-spektrum, hvilket svarer til et udbytte på ca. 70%.To a stirred ice-cooled solution of 3.3 g (0.01 mole) of 1'-ethoxy-carbonyloxyethyl-68-aminopenicillanate in 35 ml of dry ethanol-free chloroform is added 2.8 ml (0.02 mole) of triethylamine and then 1.1 g (0.01 mole) of 1,1-dichlorodimethyl ether. The solution is allowed to stand at room temperature overnight and then cooled in an ice bath, after which 1.1 ml (0.01 mole) of hexamethylenimine is added. The mixture is stored 26.145699 in the refrigerator overnight, after which the solvent is evaporated in vacuo. The residue is partitioned between ethyl acetate and water. After drying the ethyl acetate phase, the solvent is evaporated to give 3.8 g of an oil residue containing ca. 80% of the desired product, according to its NMR spectrum, which corresponds to a yield of approx. 70%.

Det urene produkt kan ikke krystalliseres, men omdannes til krystallinsk hydrochlorid med isopropanolisk hydrogenchlorid efterfulgt af tilsætning af ether. Produktet har et NMR-spektrum (D20), som svarer til det, der fås med en autentisk prøve af produktet: f.eks. 5,56 ppm, IH, C5H, d, J = 4,0 Hz og 5,64 ppm, IH, CgH, dr J = 4,0 Hz.The crude product cannot be crystallized but converted to crystalline hydrochloride with isopropanol hydrochloride followed by addition of ether. The product has an NMR spectrum (D20) similar to that obtained with an authentic sample of the product: e.g. 5.56 ppm, 1H, C5H, d, J = 4.0 Hz and 5.64 ppm, 1H, CgH, dr J = 4.0 Hz.

Eksempel 7.Example 7

Fremstilling af 2'-ethyl-hexyloxycarbonyloxy-methyl-6-hexa-hydro-lH-azepin-l-yl-methylenaminopenicillanat (fremgangsmåde-variant a) ,Preparation of 2'-ethylhexyloxycarbonyloxy-methyl-6-hexa-hydro-1H-azepin-1-yl-methyleneaminopenicillanate (process variant a),

Til en isafkølet opløsning af 18,6 g (0,05 mol) af kaliumsaltet af benzylpenicillin i 50 ml dimethylformamid sættes i løbet af 15 minutter 11,0 g (0,05 mol) chlormethyl-2-ethylhexylcarbonat opløst i 50 ml dimethylformamid. Opløsningen omrøres natten over. Reaktionsblandingen udhældes i 200 ml mættet natriumbicarbonat-opløsning og ekstraheres 3 gange med hver gang 200 ml ethylacetat.To an ice-cooled solution of 18.6 g (0.05 mol) of the potassium salt of benzylpenicillin in 50 ml of dimethylformamide is added over 15 minutes 11.0 g (0.05 mol) of chloromethyl-2-ethylhexyl carbonate dissolved in 50 ml of dimethylformamide. The solution is stirred overnight. The reaction mixture is poured into 200 ml of saturated sodium bicarbonate solution and extracted 3 times with 200 ml of ethyl acetate each time.

De forenede ekstrakter af organisk opløsningsmiddel vaskes 3 gange med hver gang 200 ml fortyndet natriumbicarbonatopløsning og 50 ml mættet saltopløsning, tørres over magnesiumsulfat og inddampes i vakuum, hvorved fås 17,0 g af en olie. Remanensen vaskes gentagne gange ved dekantering med petroleumsether og opløses derefter i ethylacetat, omrøres sammen med 1 g aktivkul, filtreres og inddampes i vakuum, hvorved fås 5,3 g af en olieagtig remanens. Tyndtlags-chromatografi (ethylacetat-silicagel) viser 3 pletter i UV-lys.The combined organic solvent extracts are washed 3 times with 200 ml of dilute sodium bicarbonate solution and 50 ml of saturated brine, dried over magnesium sulfate and evaporated in vacuo to give 17.0 g of an oil. The residue is washed repeatedly by decantation with petroleum ether and then dissolved in ethyl acetate, stirred with 1 g of activated charcoal, filtered and evaporated in vacuo to give 5.3 g of an oily residue. Thin layer chromatography (ethyl acetate-silica gel) shows 3 spots in UV light.

Der isoleres 4,7 g 2'-ethyl-hexyloxycarbonyloxymethylbenzylpenicil-lanat (Rf-værdi 0,67) ved søjlechromatografi. Produktet viser en stærk IR-absorption ved 1790 - 1740 og 1670 cm-1 på grund af henholdsvis β-lactam, estercarbonyl og amid.4.7 g of 2'-ethylhexyloxycarbonyloxymethylbenzylpenicillanate (Rf value 0.67) is isolated by column chromatography. The product shows a strong IR absorption at 1790 - 1740 and 1670 cm -1 due to β-lactam, ester carbonyl and amide, respectively.

NMR-S pektrum (CDC13): <5 = 0,70 - 2,10(m), 15H; 1,40 (s), 6Hj 3,58 (s) 2H; 4,00 - 4,40 (m) 2H; 4,32(s) IH; 5,41(d) J = 4 Hz IH; 5,62(g)NMR-S spectrum (CDCl3): <5 = 0.70 - 2.10 (m), 15H; 1.40 (s), 6Hj 3.58 (s) 2H; 4.00 - 4.40 (m) 2H; 4.32 (s) 1H; 5.41 (d) J = 4 Hz 1H; 5.62 (g)

Jx = 4 Hz, J2 = 7 Hz IH; 5,65 (s) 2H; 6,35(d) J = 7Hz IH; 7,20 (s) 5H.Jx = 4 Hz, J2 = 7 Hz IH; 5.65 (s) 2H; 6.35 (d) J = 7Hz 1H; 7.20 (s) 5H.

27 14569927 145699

Til en omrørt opløsning af 2,1 g (10 mmol) phosphorpentachlorid i 25 ml tørt ethanolfrit chloroform sættes 2,0 g (20 mmol) quinolin og efter afkøling til -15°C 4,5 g (8,8 mmol) 21-ethyl-hexyloxycarbo-nyloxy-methyl-benzylpenicillanat opløst i 10 ml chloroform. Efter omrøring i 15 minutter ved -15°C tilsættes 7,5 ml (100 mmol) n-propa-nol i løbet af 10 minutter. Blandingen holdes ved -10°C i yderligere 15 minutter, og derefter tilsættes under kraftig omrøring 25 ml mættet saltopløsning. Efter 15 minutter ved -10°C tilsættes 2000 ml petroleumsether, og blandingen holdes ved -10°C i 1 time. Herved fås 2'-ethyl-hexyloxycarbonyloxymethyl-6-aminopenicillanat-hydrochlorid i form af et gult, halvfast udfældet materiale, som anvendes direkte i efterfølgende trin.To a stirred solution of 2.1 g (10 mmol) of phosphorus pentachloride in 25 ml of dry ethanol-free chloroform is added 2.0 g (20 mmol) of quinoline and after cooling to -15 ° C 4.5 g (8.8 mmol) ethylhexyloxycarboxyloxy-methyl-benzylpenicillanate dissolved in 10 ml of chloroform. After stirring for 15 minutes at -15 ° C, 7.5 ml (100 mmol) of n-propanol is added over 10 minutes. The mixture is kept at -10 ° C for a further 15 minutes and then 25 ml of saturated brine is added with vigorous stirring. After 15 minutes at -10 ° C, 2000 ml of petroleum ether is added and the mixture is kept at -10 ° C for 1 hour. There is thus obtained 2'-ethylhexyloxycarbonyloxymethyl-6-aminopenicillanate hydrochloride in the form of a yellow, semi-solid precipitated material which is used directly in subsequent steps.

En tør opløsning af 2,-ethyl-hexyloxycarbonyloxy-methyl-6-amino-penicillanathydrochlorid i ethanolfrit chloroform afkøles til -30°C under omrøring og under tør inert argongas. Der tilsættes 2,83 ml (0,020 mol) triethylamin. Der tilsættes under omrøring i løbet af 20 minutter en opløsning af 1,76 g (0,00967 mol) [hexahydro-lH--azepin-l-yl]-chloroformiminiumchlorid i 7,5 ml tørt ethanolfrit chloroform, hvorhos temperaturen holdes ved -25°C. Blandingen omrøres under isafkøling i yderligere 30 minutter. Opløsningen inddampes i vakuum ved +20°C. Der udfældes triethylaminhydrochlorid ved triturering af remanensen med 30 ml acetone. Det udfældede materiale frafiltreres, og opløsningen inddampes i vakuum ved 20°C. Remanensen tritureres 2 gange med hver gang 50 ml petroleumsether, inddampes og opløses i 10 ml 4-methyl-2-pentanon. Efter opbevaring af opløsningen ved 3°C natten over fås 21-ethyl-hexyloxycarbonyloxy--methyl-6-(hexahydro-lH-azepin-l-yl)-methylenaminopenicillanat i form af et udfældet materiale, smeltepunkt 119 - 121°C.A dry solution of 2, -ethylhexyloxycarbonyloxy-methyl-6-amino-penicillanate hydrochloride in ethanol-free chloroform is cooled to -30 ° C with stirring and under dry inert argon gas. 2.83 ml (0.020 mol) of triethylamine is added. With stirring over 20 minutes, a solution of 1.76 g (0.00967 mole) of [hexahydro-1H-azepin-1-yl] chloroformiminium chloride in 7.5 ml of dry ethanol-free chloroform is added, keeping the temperature at - 25 ° C. The mixture is stirred under ice-cooling for an additional 30 minutes. The solution is evaporated in vacuo at + 20 ° C. Triethylamine hydrochloride is precipitated by trituration of the residue with 30 ml of acetone. The precipitated material is filtered off and the solution is evaporated in vacuo at 20 ° C. The residue is triturated twice with 50 ml of petroleum ether each time, evaporated and dissolved in 10 ml of 4-methyl-2-pentanone. After storing the solution at 3 ° C overnight, 21-ethylhexyloxycarbonyloxy-methyl-6- (hexahydro-1H-azepin-1-yl) methylene aminopenicillanate is obtained as a precipitated material, m.p. 119-121 ° C.

IR-spektrum (KBr): stærk absorption ved 1785 cm 1760 cm ^ og 1690 cm ^ på grund af β-lactam, carbonat og estercarbonyler og iminogruppen.IR spectrum (KBr): strong absorption at 1785 cm 1760 cm 2 and 1690 cm 2 due to β-lactam, carbonate and ester carbonyls and the imino group.

Eksempel 8.Example 8.

Fremstilling af ethoxycarbonyloxymethyl-63-[(hexahydro-1-(2H)-azo-cinyl)-methylenamino]-penicillanathydrochlorid (fremgangsmåde " variant a).Preparation of Ethoxycarbonyloxymethyl-63 - [(hexahydro-1- (2H) -azocinyl) methylenamino] -penicillanate hydrochloride (process "variant a).

28 US89928 US899

En opløsning af 5,4 g (0,015 mol) ethoxycarbonyloxymethyl-6 β-amino-penicillanathydrochlorid i 30 ml tørt ethanolfrit chloroform afkøles ved -40°C, og der tilsættes under omrøring 4,2 ml (0,030 mol) tri-ethylamin. En opløsning af 2,9 g (0,015 mol) [hexahydro-1(2H)-azo-cinyl]-chloroformiminiumchlorid i 7 ml tørt ethanolfrit chloroform sættes til den vundne opløsning ved -30°C og under omrøring. Reaktionsblandingen lades antage en temperatur på -5°C i løbet af 1 time, derefter udrystes opløsningen 3 gange med hver gang 115 ml iskold mættet saltopløsning og tørres over magnesiumsulfat i et isbad. Opløsningmidlet afdampes derpå under reduceret tryk, og remanensen opløses i 12 ml methylisobutylketon og lades henstå ved 5°C natten over, hvorefter der fås et krystallinsk produkt. Udbytte 2,7 g, smeltepunkt 137 - 139°C (sønderdeling), = + 204,5° (c = 1 i 96%'s ethanol).A solution of 5.4 g (0.015 mol) of ethoxycarbonyloxymethyl-6β-amino-penicillanate hydrochloride in 30 ml of dry ethanol-free chloroform is cooled at -40 ° C and 4.2 ml (0.030 mol) of triethylamine are added with stirring. A solution of 2.9 g (0.015 mol) of [hexahydro-1 (2H) -azocinyl] chloroformiminium chloride in 7 ml of dry ethanol-free chloroform is added to the obtained solution at -30 ° C and with stirring. The reaction mixture is allowed to reach a temperature of -5 ° C over 1 hour, then the solution is shaken 3 times with 115 ml of ice-cold saturated brine each time and dried over magnesium sulfate in an ice bath. The solvent is then evaporated under reduced pressure and the residue is dissolved in 12 ml of methyl isobutyl ketone and left to stand at 5 ° C overnight, whereupon a crystalline product is obtained. Yield 2.7 g, mp 137 - 139 ° (dec.) = + 204.5 ° (c = 1 in 96% ethanol).

IR-Spektrum (KBr): 1800 cm 1 (β-lactam, carbonyl), 1765 cm 1 (estercarbonyl) 1680 cm 1 (CH=fe-).IR Spectrum (KBr): 1800 cm 1 ((β-lactam, carbonyl), 1765 cm 1 ((ester carbonyl) 1680 cm 1 ((CH = Fe fe).

NMR-Spektrum (CDClg) : bred singlet ved 5,61 ppm (2H, protoner i penicillinkernens 5- og 6-stilling), dublet ved 7,91 ppm (l.H, J = 11 Hz, N-CH=fe), dublet ved 11,78 ppm (IH, J = 11 Hz, N-CH=ÉH).NMR Spectrum (CDCl3): broad singlet at 5.61 ppm (2H, protons at the 5- and 6-position of the penicillin nucleus), doubled at 7.91 ppm (1H, J = 11 Hz, N-CH = Fe), doubled at 11.78 ppm (1H, J = 11 Hz, N-CH = EH).

Eksempel 9.Example 9

Fremstilling af 1'-ethoxycarbonyloxyethyl-δβ-[(hexahydro-1(2h)--azocinyl)^methylenamino]-penicillanathydrochlorid (fremgangsmådevariant a) .Preparation of 1'-ethoxycarbonyloxyethyl-δβ - [(hexahydro-1 (2h) -azocinyl) -methyleneamino] -penicillanate hydrochloride (process variant a).

En opløsning af 0,02 mol 11-ethoxycarbonyloxyethyl-6P-amino-penicillanathydrochlorid i 40 ml tørt ethanolfrit chloroform afkøles til -40°C, og der tilsættes under omrøring 5,5 ml (0,04 mol) triethylamin. Temperaturen lades stige til -30°C, og der tilsættes under omrøring en opløsning af 3,9 g (0,02 mol) [hexahydro-l(2)~ -azocinyl]-chloroformiminiumchlorid i 10 ml tørt ethanolfrit chloroform. Efter 50 minutter er reaktionsblandingens temperatur steget til -15°C. Opløsningen inddampes derpå i vakuum, og til remanensen sættes 100 ml tørt toluen. Uopløseligt materiale fjernes ved filtrering, og filtratet udrystes 4 gange med hver gang 150 ml vandholdig kaliumchlorid-hydrogenchlorid med pH-værdi 2,2. De forenede vandfaser vaskes med 100 ml toluen, og derpå sættes 170 g natrium- 29 145690 chlorid til vandfasen, som ekstraheres 3 gange med hver gang 100 ml chloroform. Efter tørring af de forenede chloroformfaser over magnesiumsulfat i isbad afdampes opløsningsmidlet i vakuum, og remanensen opløses i 20 ml ethylisobutylketon og lades henstå ved 5°C natten over, hvorved fås et krystallinsk produkt. Udbytte 2,9 g, smeltepunkt 131 - 135°C (sønderdeling), = + 190,3°c (c = 1 i 96%1 s ethanol).A solution of 0.02 mole of 11-ethoxycarbonyloxyethyl-6β-amino-penicillanate hydrochloride in 40 ml of dry ethanol-free chloroform is cooled to -40 ° C and 5.5 ml (0.04 mole) of triethylamine is added with stirring. The temperature is allowed to rise to -30 ° C and, with stirring, a solution of 3.9 g (0.02 mol) of [hexahydro-1 (2) ~ -azocinyl] chloroformiminium chloride in 10 ml of dry ethanol-free chloroform is added. After 50 minutes the temperature of the reaction mixture has risen to -15 ° C. The solution is then evaporated in vacuo and to the residue is added 100 ml of dry toluene. Insoluble material is removed by filtration and the filtrate is shaken 4 times with 150 ml of aqueous potassium chloride hydrogen chloride at pH 2.2 each time. The combined aqueous phases are washed with 100 ml of toluene and then 170 g of sodium chloride are added to the aqueous phase, which is extracted 3 times with 100 ml of chloroform each time. After drying the combined chloroform phases over magnesium sulfate in an ice bath, the solvent is evaporated in vacuo and the residue is dissolved in 20 ml of ethyl isobutyl ketone and left to stand at 5 ° C overnight to give a crystalline product. Yield 2.9 g, mp 131-135 ° C (decomp.) = + 190.3 ° c (c = 1 in 96% 1 sec ethanol).

IR-Spektrum (KBr): 1800 cm 1 (β-lactamcarbonyl), 1760 cm 1 (ester-carbonyl), 1675 cm”1 (CH=fe-).IR Spectrum (KBr): 1800 cm 1 ((β-lactam carbonyl), 1760 cm 1 ((ester-carbonyl), 1675 cm ”1 (CH = fe-).

NMR-Spektrum (CDCl^): bred singlet ved 5,65 ppm (2H, protoner i penicillinkernens 5- og 6-stilling), dublet ved 7,89 ppm (IH, J = 12 Hz, N-CH=ÉH), dublet ved 11,88 ppm (IH, J = 12 Hz, N-CH=fe).NMR Spectrum (CDCl3): broad singlet at 5.65 ppm (2H, protons at the 5- and 6-position of the penicillin nucleus), doubled at 7.89 ppm (1H, J = 12 Hz, N-CH = ÉH), doubled at 11.88 ppm (1H, J = 12 Hz, N-CH = Fe).

Eksempel 10.Example 10.

Fremstilling af 11 (S) -ethoxycarbonyloxyethyl-6(3- (hexahydro-lH--azepin-l-yl)-methylenaminopenicillanathydrochlorid (optisk isomer).Preparation of 11 (S) -ethoxycarbonyloxyethyl-6 (3- (hexahydro-1H-azepin-1-yl) -methyleneamopenicillanate hydrochloride (optical isomer)).

A. Fremstilling af 1'(S)-ethoxycarbonyloxyethyl-6&-aminopenicillanat-hydrochlorid.A. Preparation of 1 '(S) -ethoxycarbonyloxyethyl-6β-aminopenicillanate hydrochloride.

En opløsning af 7,4 g (0,016 mol) 1'-(S)-ethoxycarbonyloxyethyl-benzylpenicillanat i 10 ml tørt ethanolfrit chloroform sættes til en omrørt blanding af 3,8 g (0,018 mol) phosphorpentachlorid og 4,8 g (0,037 mol) quinolin i 25 ml tørt ethanolfrit chloroform ved -20°C i tør nitrogengasatmosfære. Efter omrøring ved denne temperatur i 1 time afkøles opløsningen til -30°C, og der tilsættes i løbet af 20 minutter 12 ml tørt propanol. Temperaturen lades stige til -15°C i løbet af 30 minutter, hvorefter der under kraftig omrøring og under en temperaturstigning til -5°C tilsættes 25 ml kold saltopløsning. Reaktionsblandingen hældes i 50 ml iskold ligroin og omrøres i 15 minutter, derefter tilsættes yderligere 55 ml iskold ligroin, og blandingen omrøres i yderligere 10 minutter. Blandingen lades separere, og olie-vand-fasen opsamles og ekstraheres med 30 ml chloroform. Chloroformfasen tørres ved 0°C over natriumsulfat, og IR-analysen (CHClg) viser β-lactam (1790 cm 1) og ester (1765 cm 1).A solution of 7.4 g (0.016 mol) of 1 '- (S) -ethoxycarbonyloxyethyl-benzylpenicillanate in 10 ml of dry ethanol-free chloroform is added to a stirred mixture of 3.8 g (0.018 mol) of phosphorus pentachloride and 4.8 g (0.037 mol ) quinoline in 25 ml dry ethanol-free chloroform at -20 ° C in dry nitrogen gas atmosphere. After stirring at this temperature for 1 hour, the solution is cooled to -30 ° C and 12 ml of dry propanol is added over 20 minutes. The temperature is allowed to rise to -15 ° C over 30 minutes, after which, with vigorous stirring and a temperature rise to -5 ° C, 25 ml of cold saline is added. The reaction mixture is poured into 50 ml of ice-cold ligroin and stirred for 15 minutes, then an additional 55 ml of ice-cold ligroin is added and the mixture is stirred for a further 10 minutes. The mixture is allowed to separate and the oil-water phase is collected and extracted with 30 ml of chloroform. The chloroform phase is dried at 0 ° C over sodium sulfate and the IR analysis (CHCl 3) shows β-lactam (1790 cm 1) and ester (1765 cm 1).

30 US899 I NMR-Spektrum (CDCl^) ses ved 5,68 ppm IH, d, CgH, J = 4,0 Hz, og ved 5,20 ppm IH, d, C^H, J = 4,0 Hz.30 NMR Spectrum (CDCl3) is seen at 5.68 ppm 1H, d, CgH, J = 4.0 Hz, and at 5.20 ppm 1H, d, C1 H, J = 4.0 Hz.

B. I1-(S)-ethoxycarbonyloxyethyl-δβ-(hexahydro-lH-azepin-l-yl)--raethylenaminopenicillanathydrochlorid.B. 1- (S) -ethoxycarbonyloxyethyl-δβ- (hexahydro-1H-azepin-1-yl) -ethylenaminopenicillanate hydrochloride.

Den ovenfor vundne chloroformopløsning 1'-(S)-ethoxycarbonyloxy-ethyl-63-aminopenicillanathydrochlorid afkøles til - 40°C, og der tilsættes under omrøring 5,80 ml (0,042 mol) triethylamin. Temperaturen lades stige til -25°C, og derpå tilsættes dråbevis en opløsning af 3,79 g (0,021 mol) [hexahydro-lH-azepin-l-yl-chloroformi-miniumchlorid] i 10 ml tørt ethanolfrit chloroform. Blandingen omrøres derpå i 30 minutter ved 0°C. Efter afdampning af opløsningsmidlet' under reduceret tryk blandes den resulterende remanens med 50 ml tørt acetone og filtreres. Opløsningsmidlet fjernes fra filtratet, som hældes over med petroluemsether, hvorefter den sirupslignende remanens opløses under omrøring med 10 ml 2-pentanon. Opløsningen holdes ved 5°C natten over, hvorefter råproduktet krystalliseres og isoleres ved filtrering. Råproduktet opløses i 12 ml methylenchlorid og vaskes 2 gange med hver gang 15 ml kold mættet saltopløsning. Efter tørring af den organiske fase over natriumsulfat afdampningsmidlet under reduceret tryk, og remanensen blandes med 5 ml tørt acetone, hvorved fås et analytisk rent krystallinsk produkt, smeltepunkt 147 - 149°C (sønderdeling) , = + 163,5° (c = li 96%'s ethanol).The above-obtained chloroform solution 1 '- (S) -ethoxycarbonyloxy-ethyl-63-aminopenicillanate hydrochloride is cooled to -40 ° C and 5.80 ml (0.042 mol) of triethylamine is added with stirring. The temperature is allowed to rise to -25 ° C and then a solution of 3.79 g (0.021 mol) [hexahydro-1H-azepin-1-yl-chloroforminium chloride] is added dropwise in 10 ml of dry ethanol-free chloroform. The mixture is then stirred for 30 minutes at 0 ° C. After evaporation of the solvent under reduced pressure, the resulting residue is mixed with 50 ml of dry acetone and filtered. The solvent is removed from the filtrate, which is poured over with petroleum ether and the syrup-like residue is dissolved with stirring with 10 ml of 2-pentanone. The solution is kept at 5 ° C overnight, after which the crude product is crystallized and isolated by filtration. The crude product is dissolved in 12 ml of methylene chloride and washed twice with 15 ml of cold saturated brine each time. After drying the organic phase over sodium sulfate evaporator under reduced pressure, and the residue is mixed with 5 ml of dry acetone to give an analytically pure crystalline product, mp 147 - 149 ° C (dec.) = + 163.5 ° (c = l 96% ethanol).

IR-Spektrum (KBr): 1795 cm ^ (β-lactamcarbonyl), 1760 cm ^ (ester-carbonyl), 1675 cm ^ (-CH=fe).IR Spectrum (KBr): 1795 cm ^ ((β-lactam carbonyl), 1760 cm ^ ((ester-carbonyl), 1675 cm ^ ((-CH = Fe).

Et proton-NMR-spektrum for produktet viser kun et signal for C-3 protonen, hvilket antyder tilstedeværelsen af kun én stereoisomer.A proton NMR spectrum for the product shows only one signal for the C-3 proton, suggesting the presence of only one stereoisomer.

Det bekræftes ved 13C-NMR-spektrum i CDClg, eftersom der kun kan ses ét signal for 2β-methyl-carbonatornet ved 29,44 ppm (i forhold til TMS), medens en blanding af de to stereoisomerer giver signaler for dette carbonatom ved 29,51 og 30,50 ppm.It is confirmed by 13 C NMR spectrum in CDCl 3, since only one signal can be seen for the 2β-methyl carbonator at 29.44 ppm (relative to TMS), while a mixture of the two stereoisomers gives signals for this carbon atom at 29 , 51 and 30.50 ppm.

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AnalyseAnalysis

Beregnet for ^20^32^^3^6^ C 50,25; H 6,75; Cl 7,42; N 8,79; S 6,71 Fundet: C 50,21; H 6,62; Cl 7,55; N 8,89; S 6,56.Calcd for ^ 20 ^ 32 ^^ 3 ^ 6 ^ C 50.25; H, 6.75; Cl 7.42; N, 8.79; S 6.71 Found: C, 50.21; H, 6.62; Cl 7.55; N, 8.89; S, 6.56.

NMR-Analyse D20 giver følgende data: 1,27 ppm 3H, t, (J = 7,0 Hz) -OCH2CH3 r -OCHO- CH3 1.4 - 2,1 ppm 17H, m C^iCH^ ,CIT ch2 ch7 ^NMR Analysis D20 gives the following data: 1.27 ppm 3H, t, (J = 7.0 Hz) -OCH2CH3 r -OCHO- CH3 1.4 - 2.1 ppm 17H, m C

(-1 N(-1 N

CH2 / ^ CH2^ CH2 V - 3.4 - 3,9 ppm 4H, m ^C^ N-/ _2 4,25 ppm, 2H, q, (J = 7,0Hz) -OCH2CH3 4,65 ppm, 1H, s, C^H 5,56 ppm IH, d, (J = 4,0 Hz) C^H 5,64 ppm IH, d, (J = 4,0Hz) C^H 6,79 ppm, IH, q, (J = 5,5Hz) -OCH- 8.05 ppm 1H, bred s, N-CH=^-CH2 / ^ CH2 ^ CH2 V - 3.4 - 3.9 ppm 4H, m ^ C ^ N- / _2 4.25 ppm, 2H, q, (J = 7.0Hz) -OCH2CH3 4.65 ppm, 1H, s , C 1 H 5.56 ppm 1H, d, (J = 4.0 Hz) C 1 H 5.64 ppm 1H, d, (J = 4.0Hz) C 1 H 6.79 ppm, 1H, q, (J = 5.5Hz) -OCH- 8.05 ppm 1H, broad s, N-CH =

HH

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Eksempel 11.Example 11.

1'-1sopropyloxycarbonyloxyethyl-6 β-(hexahydro-lH-azepin-l-yl)--methylenaminopenicillanathydrochlorid.1'-1sopropyloxycarbonyloxyethyl-6β- (hexahydro-1H-azepin-1-yl) -methylenaminopenicillanate hydrochloride.

En opløsning af 4,65 g (0,01 mol) 11-isopropyloxycarbonyloxyethyl-benzylpenicillanat i 30 ml tørt toluen afkøles under tørt nitrogen til -20°C. Til den omrørte opløsning sættes 4,05 ml (0,05 mol) tørt pyridin og derefter 2,85 g (0,0125 mol) phosphorpentachlorid i portioner. Blandingen omrøres ved -5°C i 2 timer. Udfældet pyridin-hydrochlorid fjernes ved filtrering, og filtratet udrystes 2 gange med hver gang 50 ml iskold mættet natriumbicarbonatopløsning og derefter med 50 ml iskold mættet saltopløsning og tørres derpå i et isbad over magnesiumsulfat. Efter frafiltrering af tørringsmidlet afdampes opløsningsmidlet under reduceret tryk, hvorved fås 5,0 g af en olieagtig remanens af det tilsvarende iminochlorid. Dette opløses i 15 ml tørt ethanolfrit chloroform, og opløsningen afkøles under tørt nitrogen til -30°C, hvorefter der under omrøring tilsættes 2,14 ml 4,15M hydrogenchloridopløsning (0,009 mol) i 1,3-propandiol. Temperaturen lades stige til -15°C i løbet af 30 minutter, derpå tilsættes under kraftig omrøring i 5 minutter 3,4 g natriumchlorid i 15 ml vand, hvorefter temperaturen stiger til -5°C. Derpå tilsættes 63 ml isafkølet ligroin, og blandingen omrøres i 20 minutter. Ligroin-vandfasen fjernes ved dekantering, og den tilbageværende gummi vaskes to gange med hver gang 50 ml koldt ether og opløses derpå i 40 ml tørt ethanolfrit chloroform. Opløsningen afkøles til -30°C under tørt nitrogen, og der tilsættes under omrøring 2,61 ml (0,0187 mol) triethylamin og derpå en opløsning af 1,84 g (0,009 mol) (hexahydro-lH-azepin-l-yl)-chloroformiminiumchlorid i 5 ml tørt, ethanolfrit chloroform. Opløsningens temperatur lades stige til -20°C i løbet af 30 minutter, og derpå tilsættes under omrøring 50 ml iskold mættet saltopløsning. Chloroformfasen udrystes med yderligere 50 ml iskold mættet saltopløsning og tørres derpå over magnesiumsulfat i et isbad i 30 minutter. Efter frafiltrering af tørringsmidlet og afdampning af opløsningsmidlet under reduceret tryk opløses remanensen i 20 ml methylisobutylketon og lades henstå i køleskabet natten over, hvorved fås 0,37 g krystallinsk produkt.A solution of 4.65 g (0.01 mole) of 11-isopropyloxycarbonyloxyethyl benzylpenicillanate in 30 ml of dry toluene is cooled under dry nitrogen to -20 ° C. To the stirred solution, 4.05 ml (0.05 mole) of dry pyridine and then 2.85 g (0.0125 mole) of phosphorus pentachloride are added in portions. The mixture is stirred at -5 ° C for 2 hours. Precipitated pyridine hydrochloride is removed by filtration and the filtrate is shaken twice with 50 ml of ice-cold saturated sodium bicarbonate solution, and then with 50 ml of ice-cold saturated brine, and then dried in an ice bath over magnesium sulfate. After filtering off the desiccant, the solvent is evaporated under reduced pressure to give 5.0 g of an oily residue of the corresponding imino chloride. This is dissolved in 15 ml of dry ethanol-free chloroform, and the solution is cooled under dry nitrogen to -30 ° C and then stirred with stirring 2.14 ml of 4.15M hydrogen chloride solution (0.009 mol) in 1,3-propanediol. The temperature is allowed to rise to -15 ° C over 30 minutes, then vigorously stir for 5 minutes 3.4 g of sodium chloride in 15 ml of water, after which the temperature rises to -5 ° C. Then 63 ml of ice-cooled ligroin is added and the mixture is stirred for 20 minutes. The ligroin aqueous phase is removed by decantation and the remaining gum is washed twice with 50 ml of cold ether each time and then dissolved in 40 ml of dry ethanol-free chloroform. The solution is cooled to -30 ° C under dry nitrogen and stirred with stirring 2.61 ml (0.0187 mol) of triethylamine and then a solution of 1.84 g (0.009 mol) (hexahydro-1H-azepin-1-yl) ) -chloroformiminium chloride in 5 ml of dry ethanol-free chloroform. The temperature of the solution is allowed to rise to -20 ° C over 30 minutes and then 50 ml of ice-cold saturated brine is added with stirring. The chloroform phase is shaken with an additional 50 ml of ice-cold saturated brine and then dried over magnesium sulfate in an ice bath for 30 minutes. After filtration of the desiccant and evaporation of the solvent under reduced pressure, the residue is dissolved in 20 ml of methyl isobutyl ketone and left to stand overnight in the refrigerator to give 0.37 g of crystalline product.

Claims (3)

33 145599 IR-Spektrum (KBr): 1790 cm-1 (β-lactamcarbonyl), 1760 cm-1 (estercarbonyl), 1680 cm 1 (CH=Sh-). NMR-Spektrum (CDC13): bred singlet ved 5,57 ppm (2H, protoner i penicillinkernens 5- og 6-stilling), bred singlet ved 7,80 ppm (IH, N-CH=fe).IR Spectrum (KBr): 1790 cm -1 (β-lactam carbonyl), 1760 cm -1 (ester carbonyl), 1680 cm -1 (CH = Sh-). NMR Spectrum (CDCl3): broad singlet at 5.57 ppm (2H, protons at the 5- and 6-position of penicillin nucleus), broad singlet at 7.80 ppm (1H, N-CH = Fe). 1. Analogifremgangsmåde til fremstilling af forbindelser med den almene formel I (CH2)n N - CH = N - CH - CH 3 I I r5 CO— n- CH - COO-CH-O-COO-R4 eller terapeutisk acceptable syreadditionssalte deraf, i hvilken formel 4 n er et tal 3 - 7; R betegner en alkylgruppe med 1-8 carbon- 5 atomer; og R betegner hydrogen eller methyl, kendetegnet ved, at a) et reaktivt derivat af en forbindelse med den almene formel III (C®2^n N-CH=R6 III c. hvor n har den ovenfor anførte betydning, og R er -O- eller -S-, omsættes med en forbindelse med den almene formel IV 34 145699 HpN - CH -An analogous process for the preparation of compounds of general formula I (CH 2) n N - CH = N - CH - CH 3 II R 5 CO - n - CH - COO-CH-O-COO-R 4 or therapeutically acceptable acid addition salts thereof, in which formula 4 n is a number from 3 to 7; R represents an alkyl group of 1-8 carbon atoms; and R is hydrogen or methyl, characterized in that a) a reactive derivative of a compound of the general formula III (C2-2 n N-CH = R6 III c. where n is as defined above and R is - O- or -S-, is reacted with a compound of the general formula IV 34 145699 HpN - CH - 1. I ch3 f CO—N-CH - C00-CH-O-COO-R4 IV '4 5 hvor R og R har den ovenfor anførte betydning; eller b) en forbindelse med den almene formel V ^^ ,0 - R7 (CH2>n /-CH\ \ - r7 V hvor n har den ovenfor anførte betydning, og R betegner en alkylgruppe, omsættes med en forbindelse med den almene formel IV /Sx ^CHX H~N - CH - CH NC^ ^ * i \ _5 I I CH3 i CO—N-CH - COO-CH-O-COO-R4 IV 4 5 hvor R og R har den ovenfor anførte betydning; eller c) et reaktivt derivat af en forbindelse med den almene formel VX ^\ /CH3 HCONHCH - CH NC I R5 VI I 5 [_ CO—N-CH - C00-CH-O-COO-R4 4 5 hvor R og R har den ovenfor anførte betydning, omsættes med en forbindelse med den almene formel VII NH VII hvor n har den ovenfor anførte betydning; eller1. In CH3 f CO-N-CH-C00-CH-O-COO-R4 IV '45 where R and R are as defined above; or b) a compound of the general formula V 1, O - R 7 (CH 2> n / -CH 2 - R 7 V where n is as defined above and R represents an alkyl group, is reacted with a compound of the general formula IV / Sx ^ CHX H ~ N - CH - CH NC ^ ^ * i \ _5 II CH3 in CO-N-CH - COO-CH-O-COO-R4 IV 4 5 wherein R and R have the meaning given above; or c) a reactive derivative of a compound of the general formula VX 2 / CH 3 HCONHCH - CH NC I R 5 VI I 5 [-CO-N-CH - C00-CH-O-COO-R 4 4 wherein R and R has the meaning given above, is reacted with a compound of the general formula VII NH VII wherein n has the meaning given above; or