DE2155081A1 - New heterocyclic compounds - Google Patents

Description

Dr. F. Zumsteln ε & »\. - Dr. E. Assmanr »Dr. R. Koenlgsberger - Dlpl.-Phys. H. Hotebauer - Tue. F. Zumsteln Jun.

PATENT LAWYERS TELETON COLLECTION NO 22 5341

TELEX 529979

TELCGRAMME ZUMPAT CHECK ACCOUNT MUNCHE-N 91139

BANK ACCOUNT BANKIIAUS H HOUSES

8 MUNICH 2.

BRÄUHAUSSTRASSfc 4 / IiI

DUI 1166

KOEIwKLlJKE BEOERLANDSCHE GIST- EN SPIRIYUSFAPRIEK N

DeIf t / NIEDiJRLAlJDE

"New Heterocyclic Compounds"

The present invention relates to new therapeutically useful penicillanic acid and cephalosporanic acid derivatives, Process for their preparation and pharmaceutical compositions containing these compounds.

The new penicillanic acid and cephalosporanic acid derivatives according to the invention are connections of the following general Porrael 1

2 -

209820/1159

BADORiGiNAi.

- CCi _o »R ^ O

H
NQ

(D

wherein Q is a group of the formulas

H
-C

O'

.H
^ COOH

(II)

or _c . 0 *

-C '

CII ₀ X

COOH

(III)

represents

where X is an atom, a Hydro Vyg.'uppe or a Means lower alkanoyloxy group or the group -CHpX and the carboxyl group of the formula III with the formation of a lactone group, i.e. a group of the formula

-CO-CH ₂ -

Il

or with formation of a lactan group, i.e. a group dcr formula

-C-NH-CH ₀ -0

209820/1159

are connected, E is a lower alkyl group or an aryl group, e.g. a phenyl group or a haphthy! group, the optionally one or more substituents, such as Chloratoiae, Fluorine atoms, nitro groups, amino groups or i-lower alkyl groups carries (preferably the 2,6-dichlorophenyl group represents) or a tertiary alkyl group such as the adamantyl group. R is a hydrogenator or an alkyl group or a carboxyl group optionally esterified with a lower alkyl group or one in a Alkali metal salt, an alkaline earth metal salt or an amine salt converted carboxyl group, a Carbaray! group, a

2 cyano group, an amino group or a chloratora, R a

Hydrogen atom, a cyano group, an amino group, a NiecU'igaralkoxycarbony! Amino groups, a lower alkyl group or one with a lower alkyl group, an iry "I group (preferably a phenyl group) or an aralkyl group

(preferably a benzyl group) substituted carboxyI-

group, or in which R is a carbamoyl group which optionally carries a lower alkyl group or a Ph.eny! - (Lower) -alkyl-mononalogensilyl esters, the benzyl esters or the phenacyl esters) and the amides (for example the SaccharyIderivate) of the penicillanic acids and cephalosporanic acids mentioned. The term "low * ¹ , as used in the present specification in relation to the alkyl or alkanoyl groups, means that the group contains at most 4 carbon atoms.

The compounds according to the invention can be prepared by several different processes, each of which involves an application of a known process for the preparation of penicillin on and cephalosporins !! represent.

215508t

Erf indungsgemäS w # * 4en CFIE compounds of the general formula prepared by "in a salt, an ester or an amide of a 6-or 7-Aminopenicillansäure- Aaiinocephalosporansäure compound of the formulas:

H
H ₂ N - CC

0 '

-N-

(IV)

or

H / C

II - Ν

(V)

CH,

C-CH ₉ X

COOH

wherein X has the meaning given above, wherein the substituent X, when it represents a hydroxyl group, is preferred is protected with an active Enter (e.g. «a 2.4" * Dinitrophenyl ester, a p-Nitrophenylester or an N-Hydroxysucclnirnidester) an acid of the general formula VI

H
-C- COOH

(VI)

1 2
in which R, R ¹ and R d each have the meanings given above, or with an active funct3o »^ llen 3> derivative which is suitable as an acylating agent for a primary amino group, for which reason.

209820/1159

215508t

esters chloride, CürbonsäurobroiKide, Säureanhydriüe including from stronger acids, such as lower aliphatic carbon säureciono ^, alkyl or aryl acids and sterically hindered acids "as dip" henylessigsäure mixed Anhydridο produced a.

In addition, an acid azide or an active thioester (e.g. with thiophenol or thioacetic acid) of the acid can be used.

Alternatively, the free acid itself can using a CarbcM imide Beaktionsteilnehmers with the 6-Aminopenicillonbäure- or 7-aminocephalosporanic acid compound are coupled Anstell _o? the 2,4-dinitrophenyl and p-uritrophenyl esters can be a corresponding azolide, ie an amide of the corresponding acid, the amide nitrogen of which is part of a quasi-aromatic 5-membered ring which carries at least two nitrogen atoms, ie imidazoles, pyrazoles, triazoles ,, benjimid & zol, benzotriazole and the substituted derivatives of these compounds can be used.

The procedures for carrying out these reactions with education of a penicillin or a cephalosporin and the methods for isolating the compounds so prepared e.g. known from British patents 932,644, 957 570, 959 054, 952 519, 932 530, 967 108 and 967 890.

The product obtained by the above-mentioned process in the form of the ester, the roller or the amide can be converted into the corresponding Penieillanufiui'c) or into the corresponding cophalosporanic acid according to procedures known per se Ti'ialkylrnlyloi.toj . becomes, the vereiitßXTiite XtiHüpp ^ easily forming the

corresponding acid of the general formula I are hydrolyzed.

Another inventive method for producing the Compounds of the general formula I consist in that an acid of the general formula A-COOH (in which A is a group of the following formula:

RR ¹

H -C-

R ² (VII)

1 2

means in which R, R and R have the meanings given above and the reactive 'suitably protected in the radical A Has groups, with a 6-isocyanatopenicillanic acid or a 7-isocyanato (desacetoxy) cephalosporanic acid compound of the formula O = C = N-Q (v / orin Q has the meaning given above), which have atoms or groups which the carboxyl group and optionally the hydroxyl group when present, i.e. when the group X is in the Formula III means a hydroxyl group, protect, reacts «preferably lot the group containing the carboxyl group or any existing liydroxygrunpo dec G-lsocyauatopenj cillansäiire- or T-IcocyanatücophaloiiporiincaiirG Reaction Part No. protects, a di- or trialkylsilyl group, which are easily produced by hydrolysis of <3t. ". i Product can be separated / grounded.

The reaction of the carboxylic acid of the formula A-COOH with dom ir.ocyanate der! «Oriael OGIJ-Q is preferably used in an inert

209820/1 159

Organic LofruncsiBitteliaedlum, such as toluene, chlorothane or Bensonitrlit & ^ rcb ^ £ u "hri, a small amount of an organic base, & .S. a substituted iridazole itstei serve as a catalyst Penicillans & ureoerivaU specified Heaktionaschema

_V A - COOH l 0-C = JJ-CH - CH C ^y * A-CO-NH-CH - CH C,

, Q ^ H GH ^ CN CH - 000K

COOE °

where E is the carboxyl group while R - s3i ^J. .on see (represents a doing group which is separated after the reaction, for example by hydrolysis.

A further process for the preparation of the penicillic acid and CepKiloaporansäurederivate of the a31ceiaeinr; i Forrael I consists in that nan an-isocyanatopenicillnnsäure- or T-Iöocyanatoeophalosporans.Äure-Vßrbindting dex-Forrael OsCselik-Q (where Q has the meaning given above), whose carboxyl groups or optionally present hydroxyl groups are suitably geseütsi, iait a metalX-organic compound of the formula A-Me »A-Ue-Hal or Λ-Me-A (where A has the meaning given above) _t Hc a metal atom, eg denotes a lithium, katrium or 14agneciura atom, the number I or II indicates the value of the metal and IJaI means a halogen atom (preferably a Oh'Jor- oävr Broiii-Atora), ussurtst, urj nia to separate dorr hotallions and dvr the carboxyl / 5r «i>i> e schUizonde liardrolyuiprbai'e 'group <> a« obtained ÄVfisclicnprödulct hydro-

lyoicrt. The reaction v / ircJ in an anhydrous organic Solvent riedium unier conditions which cause a reaction of the Grignard type, Reformatsky type, or a reaction more like it Favor kind, filled through.

The iGocyanatausßangRmaterialicn of the general formula O = C = Ii-Q (where Q has the meaning given above) can be obtained by reacting phosgene with a penicillanic acid or cephalosporic acid derivative of the general formula WNQ, where W is a hydrogen atom or a such group means that the group W-KH- can easily be formed by reaction ml ⁺ . Phosgene can be converted into an isocyanate group, the group Q having the meaning given above and being. the carboxyl groups and any hydroxyl groups present are suitably protected. The group W of the starting material can be introduced into the 6-aminopenicillanic acid or 7-aminocephalosporanic acid derivatives at the same time as the protective group for the carboxyl group and the hydroxyl group are introduced. Preferably, the group W is a tri- (lower) alkylsilyl _o When the group W is a lelclrti separable group, the reaction of such compounds proceeds with phosgene under the same conditions smoother than is the case when the group W represents a hydrogen atom. The reaction with phosgene must be carried out in an organic solvent medium which is inert with regard to the reactivity of the isocyanate group formed.

Toluene and methylene chloride or mixtures of these solvents are particularly suitable. Furthermore, to facilitate the reaction, an organic base can be used to bind the formed hydrogen chloride can be added * Preferably this base is a tertiary amine, such as triethylamine, which does not react with the isocyanate group. Because high temperatures

209820/1159

lead to a decomposition of the Penicillansäurc- or CEPHALOSPORANIC ACID skeleton v / ould, the reaction is preferably in β ear low temperatures, preferably at ⁰ C -4o performed.

The substituted isoxazol-5-yl-acetic acid - Auö £; angsmaterial of the general formula VI, which are essentially new and thus a further subject of the present invention Representing the invention were prepared according to the following procedures:

209820/1159 BADOBiQJNAL

ΛΟ

ο ο ο

CvJ W O

- XT-

ti 03

C \ J

¹ VS

W. O

ι ο

III O

pci

=, J

O O O OJ

CJ III O

I H

tr;

CV

Vi

CD

CAJ

(M ti; ο

O III O

W)

0)

III

CJ

209820/1159

BADOWGfNAL

The starting nitrile oxides can be used according to the well-known von Grundmann, Qujlico et al., Synthesis 1970. 341 and Exporienta £ 6, 1169 (1970) and the processes cited therein.

The reaction with n-butyl lithium in the presence of tetramethylenediamine (TMEDA) can take place in aprotic solvents such as toluene or tetrahydrofuran. The Y / eg 1 allows the greatest possible variation in the desired connections. In certain cases, the group R has been to, the 1,3-dipolar addition reaction but before treatment with butyllithium in a different Orr, PPE, _e z transferred B. -000H in the group -COMHp or -CM or -HH _2, 7.U to avoid the synthesis of the starting acotylenes, which are difficult to make in certain pills.

A group R ^ H of the formula VIA can be introduced directly via the V / eg 1 by using 1-butyne (R ² = CH ~) instead of propyne.

The other procedures are as follows:

1) a-Halogenation of compounds of the formula VIA, optionally followed by a reaction with a nucleophilic agent. For example, the ct-amino acid (Formol VI: R = 2,6-dichlorophenyl, R ¹ = H, R ² = NH _g ) via the cc-bromination of the corresponding methylisoxazole-5-γΙ-acetate with 1,3-dibromo -5,5-dimethylhydantoin, followed by hydrolysis and reaction of the α-bromic acid with concentrated ammonia. This procedure is an improved synthesis analogous to that of ibotenoic acid described in Cherc.Pharm.Bull. JU, 89 (1966).

209820/1159 BADOWQINAL

2) By treatment with Idthiuia introductory agents in the α-Position of the carboxylic acids of the formula TIA and reaction with a suitable means »

Examples of the new compounds of the general Pormol VI, which are prepared according to the scheme given above are the compounds of this formula in which the sub-

1 2

substituents R, R and R have the meanings given below own·

209820/1159 BATH ORIGINAL

OD Kl

cn to

R. Adamantyl R ¹ H R ² H Br 4-nitrophenyl H CH ₃ H H 4-aminophenyl H H K H 2,6-dichlorophenyl H H - NH - C - O - CH ₅ ~ <\ / V-NO ₀
ti w \\ // cL NH ₂ H CH ₃ 0 ^N ' H 2,6-dichlorophenyl Cl Cl 2,6-dichlorophenyl H NH ₂ CH ₃ 2,6-dichlorophenyl - g - NH ₂ H 2,6-dichlorophenyl - C £ N H 2,6-dichlorophenyl H 2,4,6-trimethylphenyl 2,4, o-trinethylpheny1 2,4,6-trimethylpheny1 2,6-dichlorophenyl
* 2,6-dichlorophenyl

cn cn ο

The new penicillanic acid and cephalosporanic acid der: j.vate of the general formula I have antibiotic properties, which, alone or mixed with other antibiotics, make them potentially useful for humans and animals. Some of the new compounds of general formula I have effects that are comparable to those of penicillin G. are. They have special effects against gram-positive organisms and continue to show a good effect against Penicillin-resistant staphylococci, especially those compounds in which R is the 2,6-dichlorophenyl group,

1 2

R is a hydrogen atom or a methyl group, R is a hydrogen atom and Q is a group of the formula II or III and in which X is an acetoxy group, as well as αIe 3? .Lze of these compounds.

The compounds according to the invention are voiisugr * veise in In the form of a non-toxic salt, such as in the form of the sodium, potassium or calcium salts for therapeutic purposes Purposes. Other salts that can be used include the nontoxic ones suitably crystallizing Salts with organic bases such as amines, e.g., trialkylamines, procaine and dibenzylamine.

In the treatment of bacterial infections, the inventive antibacterial compounds both topically, orally and parenterally appropriate for the administration of antibiotics administered in the usual way. They are administered in unit doses containing an effective amount of the active ingredient in combination with suitable physiologically compatible carrier materials or binders contain. The dosage units can be in the form of liquid preparations, such as solutions, suspensions, dispersions or Emulsions or in solid form as powders, tablets, capsules and the like.

209820/1159

Accordingly, the present invention also encompasses therapeutic ones Zusamroensetatmgeii, which is an effective amount of one of the compounds according to the invention in combination with a suitable one contain inert carrier material or binder. Such therapeutic compositions can additionally to one of these compounds also contain one or more additional therapeutic ingredients.

The expression “effective amount” as it is used in relation to the described prebindings describes an amount which, when administered in the usual way, destroys or inhibits the growth of the microorganism in question or, in other words, does not Amount to keep the growth of the bacteria under control. The size of an effective amount can be readily determined by those skilled in the art according to standard procedures for determining the relative activity of antibacterial agents when used against particular organisms by the various routes of administration.

Suitable carrier materials and binders can be any suitable physiologically acceptable ingredients which serve to facilitate the administration of the therapeutically active compound. The carrier materials can have an additional effect and can serve as a diluent, as a flavoring agent, as a binding agent, as a means to delay the effect, as stabilizers and the like.

Examples of carrier materials are water as gelatine, May contain gum arabic, algenates, dextran, polyvinylpyrrolidine, sodium carbosymethyl cellulose or the like, aqueous ethanol, syrup, isotonic saline, isotonic glucose, starch, lactose or other suitable materials,

209820/1159

commonly used in the pharmaceutical and veterinary industries

Another object of the present invention comprises a method for inhibiting the growth of bacteria by applying an effective amount of the invention antibacterial compounds on the site of bacterial attack.

For example, the method can be used to treat bacterial infections in animals by adding a <-; o An effective amount of an antibacterial compound is administered to the host animal.

The present invention is illustrated by the following examples further explained.

209820/1159 BAD OR] QlNAl

example

Preparation of 6 - {- [3- (2,6-dichlorophenyl) -isoxazol-5-yl] -acetamidoj-penicillanic acid (Sodium salt)

N - C - C

- IJ

COOlTa

In a three-necked flask equipped with a gas inlet tube, a thermometer and a dropping funnel, 755 mg (3.5 mmol) of 6-aminopenicillanic acid is suspended in 10 ml of ethyl acetate under a nitrogen atmosphere. The grater is then cooled in an ice bath and 0.51 ml (3.8 mmoles) of triethylamine are added, followed by the addition of 0.48 ml (3 »8 mmoles) of trimethylchlorosilane after 10 minutes. The mixture is stirred for about 35 minutes and then a further 0.51 ml (3.8 mmol) of triethylamine is added. Then 3- (2,6-dichlorophenyl) -isoxazol-5-yl-acetyl chloride (obtained by reacting 3- (2,6-dichlorophenyl) -isoxazol-5-yl-acetic acid with thionyl chloride in diethyl ether is added dropwise track dimethylformamide) in 5 ml of ethyl acetate to the reaction mixture in a 'such speed that the temperature does not rise above 5 ⁰ C. After the addition, the ice bath is removed and the mixture is stirred for a further 90 minutes at room temperature.

The reaction mixture .7Ir (I then under ice-cooling into a mixture of 20 ml v / water and 20 ml diethyl ether added, keeping the pH at 6.8. The aqueous layer becomes

209820/1159

BATH ORIGINAL

washed once more with 30 ml of diethyl ether. The aqueous layer is acidified to a pH of 1.5 after the addition of 40 ml of diethyl ether. After separation, the aqueous layer is washed again with 30 ml of diethyl ether. The organic layers are combined and washed once with 20 ml of ice water acidified to pH 1.5 and then with 20 ml of ice water. After drying and treatment with Norit, the organic layer is concentrated to about half its volume and then Hatrim-OL-ethylcaproate is added. The precipitated sodium salt is filtered off, washed with diethyl ether and dried. The yield of the compound, which is pure according to thin layer chromatography, is 550 mg (32 ° />) .

Example 2

Production of 6-f- [3- (2,6-dichlorophen _v ^ir l) -isoxazol-5-yl] -acetamidol-penicillanic acid (sodium salt)

A 250 ml three-necked flask is fitted with a thermometer, a good condenser and a dropping funnel. The reaction is carried out under nitrogen and 220 ml of dichloromethane and 2.72 g (10 mmol) of 3- (2,6-dichlorophenyl) -isoxazol-5-yl-acetic acid are added to the vessel. After the addition of 0.13 ml of N-vinyl imidazole as the catalyst, a solution of 3Þ g (10 mmol) of trimethylsilyl-6-isocyanatopenicillanat in dichloromethane was added dropwise to the solution kept at 20 ⁰ C stirred solution. After 23 hours the reaction is complete and the isocyanate has been converted into the desired product to an extent of about 70 °. The reaction mixture is poured into ice water buffered to a pH of 7 and extracted twice with diethyl ether. The aqueous layer is on a

209820/1159 6 ORIGINAL

Acidified pH 4.0 and extracted three times with diethyl ether. The desired product is completely separated from the aqueous layer. The collected organic layers are washed with a small amount of ice water and then dried over anhydrous magnesium sulfate, filtered and ^{concentrated at 0} ° C. to some extent in vacuo.

A solution of sodium ct-ethylcaproate in ethyl acetate is added dropwise to the concentrated solution. The colorless precipitate which forms is collected on a piIter } washed with diethyl ether and dried in vacuo. Yield 2.81 g (57 #). Analysis of the NMR spectrum of the product dissolved in hexadeuterodimethyl sulfoxide 60 Mc, tf values in ppm, tetramethylsilane as internal Standard) resulted in the following:

NH multiplet at 8.95 (0.7 protons)

C ₆ H, about 7.5 (3 protons)

Isoxazolyl-H 6.50 (proton)

-CH ₂ and ^C 2 " ^H ^ i 99 (3 protons)

Cc-H and C / - H multiplet 5.50 to 5.30 (2 Pro ^{5 ö} tone)

C ₅ - (CH ₅ ) ₂ 1.62 and 1.52 (doublet 6 protons)

Partial analysis of the IR spectrum (KBr disk, values in cm "" ¹ ):

3355 NH

1755 C = O lactam

1700 C = O amide

1610 C = O carboxylation

1505 NH deformation vibration

1600 C = C aromatic

1330 iGOxazolyl ring

755) C-G-L stretching

209820/1159

OJ ₁

I I!

Ο- ⁰ 'Ν _{β <}

In a three-necked flask with a gas inlet tube, a $ feöiia © iaiif # * üttd feifiifti $ r © t> f1: riifh $ iir gilded ir§, m & pmaMv man 500 ffig (t, 8 milol)? -Aminocephalocporatic acid iff 10 ml of ethylacetate under a nitrogen atffionsphere "Until Suepcntiion, iti öincto SifeWet gfekühit tihti aaisfi wfei? deß 0.5 C ^ i 2 ajMöi)! rriäth ^ iÄmift ^ geö #% gt * teök 5 Mitiüte" Oj 3 fflittiöiyl t müchfl and then stirred for 1 hour at room temperature.

MiöehtJiig Witd -. «Ttietit cooled and after the iJ? läthylaailiiaq. \ ilvalt! fttö becomes 3 «- (2 | 6

S-yi-fteetylchlotid (lh of \ i? Elsö as described in Example 1) in 5 al Äthylacetai iropferiWöice mi AEEF ReaktionsMöchung given woböi "stops the below 5 ⁰ O"

After the addition, the egg bath is removed and diö ffiißöhung for a further 2 hours at room temperature The reaction solution is then stirred into a mixture of

209820/1159

BATH

Pour 30 ml v / ater and 30 ml diethyl ether while cooling with ice, while keeping the pH at 7.0. 'The aqueous Layer is washed with a further 30 ml of diethyl ether and 30 ml of ethyl acetate. "After the addition of 50 mX ethyl acetate the aqueous layer is acidified to a pH of 1.7. The layers are separated and the aqueous layer is extracted again with 50 ml of ethyl acetate.

The combined ethyl acetate layers are applied once acidified ice water to a pH of 1.5 and washed twice with ice water. After separation, drying over Magnesium sulfate and treatment with Norit, the ethyl acetate layer is concentrated to about 1/3 of its volume and then sodium CT ethyl caprohate is added. The precipitated one The sodium salt is washed once with ethyl acetate and twice with η-hexane and, after filtration, dried in vacuo. The yield is 438 mg (0.8 mmol = $ 44). According to thin layer chromatography, the compound is pure.

A partial analysis of the IR analysis of the end product (KBr disk, Values in cm ") resulted in the following values:

- 3430)
ί 3280) 1670 NH = 0 ester 1760 C = 0 ß-Lac tarn and C 1690 - C = O amide 1600 C = O carboxylate ion 1558 C = C or C = N 1230)
1025) C-O-C esters 782 C-Cl

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BAD ORfQ) NAL

~ 22 -

Analysis of the NiIR spectrum of the final product j solved in Hexadeuterodimethylsulfoxide (60 Mc, cf-Vcrte in ppm, Ietramethylsilan as an internal standard) resulted in the following:

CO -

NS

O CH ₂ -C 0 -CH ₂ C ₆ -H C ₇ -H

Isoxazole-C.-H C / -H,

2.3 3.07

3.71 (AB quortet (Yes * 17.5 cps, 2 protons)

3.98 (2 protons)

4.73 - 5.20 quartet (J «=? 12 cps, 2 protons)

4.98 and 5.05 doublet (J ^ 4.5 cps ,, 1 proton)

5.47 -> 5.66 O ^ ai ^ ott (J ^ 4, ^r : cps, J '^ 8 cps,

1 proton)

6.51 (1 proton)

7,: "> 5 sharp, narrow gap pattern (3 protons)

9.22 and 9.35 doublet (J · «8 cps, 1 proton)

Elemental analysis C ₁₂ • H ₁₆ M ₃ O ₇ SCl ₂ Well 1
2 H ₂ O • found 44.97 - middle ber 26th C. 3.31 - 45.03 ok 45, 08 H 7.70 - 3.34 ok 3, 54 N 5.70 - 7.71 ok 7, 75 S. 5.68 ok 5, - $ 45.10> 3.38 io - 7.72 io P, D / / O

ber. (with 1/2 MdI crystal water)

209820/1159

BAD QKfGINAL

iff MafeiiiiBöfei ^ iis 164 1 *

0 Ii M

OOOiia

E.g. fine suspension of 620 m $ (?, B.

In 10 ösl Ith ^ laö ^ ti the ift eiriött §Ö ffll

and the eleventh üteepitfltshteii iet _t under Btiök-

Yorllet j and otichaniticli getUhübi Wi? A _f gifet n & ch d ^ fti KUhlnn der SuöpetJ ^ ioii ö'i% eihefti liebad 0,46 (5 ₍ 3 isMöi) <ctinthylaain «Hboh 5 Hinuien Vietiolämi Ö _t 4 ^ ml (Si3 ffli Vietiolämi Ö t 4 ^ ml) ftritoctliyiehlor & ilan äugüeetkt «nd öae Μίΐΐϊο« Virö during a night htie external cooling foftgöfäetiit «Then the ftöaktiottöiofchung is t? rue" t with a Siö bath tand & b e eetat "oil (2 # 9 nälUthuyla) 5- {2 | 6-dichlorophenyl) * isö3taBol * 5-yl-soetylchlori (5 (obtained according to the Yloise described in Example 1) in 5 ml of ethyl acetate is added dropwise to the reaction mixture at such a rate κυ that the flow rate does not exceed 5 ⁰ O rises »The cooling bath is then removed and stirring continued for a further 2 hours»

BAD ORtQiNAU

«24 -

The reaction mixture is then added, with cooling and mechanical stirring, to a mixture of 20 ml of water and 20 ml of diethyl ether, while keeping the pH at 7.0. The aqueous layer is washed once with 20 ml of ethyl acetate and once with 20 ml of diethyl ether. After adding 40 ml of ethyl acetate to the aqueous layer, the pH is brought to 1.7. The aqueous layer is extracted again with 30 ml of ethyl acetate. Then these layers are combined, washed once with 20 ml of acidified ice water (pH 1.7) and once with 20 ml of normal ice water. After drying over magnesium sulfate and treatment with liorite, the organic layer is concentrated to about 1/3 of its original volume. Then Nf.triuin-ct-ethyleapronate is added, the precipitated sodium salt is collected on a filter, washed with Ätl ylar-etat and with diethyl ether and dried in vacuo. The yield is 0.603 mg (1.23 mmol = 43 ° β>) . According to thin layer chromatography, this compound is pure.

A partial analysis of the IR spectrum of the end product (KBr disk, values in cm ") gave the following values:

£ 3400 NH (wide absorption)

1750 C a Oβ lactam

1670 C = O amide

1590 C = O carboxylate ion

1555 C = C

i 1540 NH def. (Shoulder)

781 C-Cl

Analysis of the NMR spectrum of the end product, dissolved in hexadeuterodimethyl sulfoxide (60 Mc, cf values \ n ppm, tetramethylsilane as internal standard) gave the following:

C ₃ -CH ₅ 1.98 (3 protons)

2.95 - # ■ 3.65 AB-quartet (J ^ 17.5 cps;

2 protons)

209820/1159

BATH ORIGINAL

CH ₂ C

C ₆ -H and

C ₆ H

₆ H ₅

- 25th

(2 protons)

4.88, 4.97 5.43-5.52

^ 4.5 cps; 2 protons) and

6.50 (1 proton)

7 »55 (sharp narrow splitting pattern; 3 protons)

example

Preparation of 6-i- [3 »(2,6-] dichlorophenyl) -4-carboxyisoxaiiol-5-yl.! - acetamido3 Pen.io.illansu.ure

COOH

N, C-CH

2 HH CNC

C N-

COOH

In a three-necked flask fitted with a gas inlet, a thermometer and a dropping funnel, 314 Wj ( ¹ > 0 mHol) trimethylsilyl-b-isocyanato-penicillanate and 316 mg (1.0 mmol) 3- ( 2,6-dichlorophenyl) -4-carboxyisoxazol-5-yl-eusic acid (obtained by the reaction of 3- (2,6-dichlorophenyl) -4-carboxy ~ 5-methylisoxa2; ol, which is produced by the 1,3-dipolar Addition of 2,6-dichlorophenylbenconifril oxide and trimethyli3.ilyl-2-butynoate formed with 2 equivalents of n-butyllithium and 1 equivalent of tetraethylethylenediamine in toluene, followed by carbonylation with CO2)

209820/1159 BAD ORTQfNAC

in 25 ml of benzonitrile · A solution of 145 kg (1 »1 ffiliol) of N-methylbenzimidazole in 5 ml of benzonitrile is added dropwise to the mixture. J ¹ In results in a direct formation of carbon dioxide. After 2 hours the evolution of carbon dioxide ceases and the reaction mixture is poured into a mixture of 30 ml of water and 50 ml of diethyl ether while cooling with ice, while maintaining the pH at 7. The aqueous layer is extracted twice with 50 ml of diethyl ether.

After 50 ml of diethyl ether and 10 ml of ethyl acetate have been added, the pH is brought to 4. The layers are separated; v.nd the aqueous layer is extracted twice with 50 ml of diethyl ether.

The combined organic layers become ice water washed and dried over magnesium sulfate. After removal of the solvent, 136 mg of a pale yellow solid are obtained which, according to thin-layer chromatography, is pure was.

A partial analysis of the IR spectrum of the end product (KBr disk, Values in cm "") resulted in the following values;

1775 C = Oβ-lactam 1700 C = O carboxyl 1600 C = C aromatic 1560 C = N 1430 Isoxazole ring 780 C = Cl

209820/1159

" ²⁷ⁱ 21S5081

The analytics of the HtfR-SpefctarciBe da »end product _t dissolved in ilexädeutcfcöäiiiieirhyltnjlföxyd (60 Mc, cT-Wejetfc in PpBi ₄ Selra iuöthylöilart as internal standard) cr ^ from ÄiMs itel & etiäe ί

C * - QiL%, $ 2 vtnu 1.65 (6 i ^ gions)

C ₅ -H and C ₆ -H 5,33 - * 5 _f 70 «ttltäplett (2 protons)

C ^ H,? »55 ßcbarfos enges

UH 9.10 J tray

Example 6

Manufacture of the natritiüjßaizes dor 6-C-C3-I

JC - OH ₉ - C * - »- CH — GH 0 (0U _x )

In the main procedure described in Example 2 1.23 g (5 »mol) of 3- (2,4,6-trimethyl) -phenyl · jooxazol-5-yl-eBBigEiiure with 1 ^ 57 g (5 liters of oil) sriinethyleilyl-6-isocyanato-penicillanate in 2fj jal dry dichloi? aethane in the presence of about 0.05 ial-vinyliraidazole as a catalyst The reaction was complete in about 6.5 hours.

209820/1159

According to thin-layer chromatography, about $ 60 of the isocyanate had been converted into the desired product. The reaction product was treated in the usual way. At a pH of 4 * 5, the penicillin was extracted from the water with diethyl ether. The ether solution was washed with a small volume of ice water, treated with activated charcoal, dried over anhydrous magnesium sulfate and slightly ^{concentrated in vacuo at 0} ° C.

A solution of sodium ct-ethyl caproate in diethyl ether was added dropwise to the concentrated solution. The colorless precipitate formed was collected by Piltrat.oii and washed repeatedly with cold diethyl ether. After drying in vacuo, 0.8 g of the product was obtained good purity of the product was indicated by thin film chromatography, by IR and by NMR spectra.

Analysis of the NMR spectrum of the end product, dissolved in an approximately 2: 1 mixture of hexadeuterodimethyl sulfoxide and D ₂ O (60 Mc, cf values in ppm, internal standard 2,2-dimethylsilapentane-5-sulfonate) gave the following values ;

C ₆ H ₂ 6.95 (singlet, 2 protons)

Isoxazolyl C.-H 6.3 (1 proton) C ₅ -H and C ^ -H about 5.45 (AB-quartet, ί ^^ Ο, Ι ppm,

J ^ g «4 cps, 2 protons)

C ₂ -H 4.2 (1 proton)

CH ₂ -CO- 3.95 (2 protons)

P-CH ₅ 2.25 (3 protons)

(o-CHj) ₂ 2.05 (6 protons)

J ₂ 1.5 and 1.6 (6 protons).

209820/1159

BATH

example

Production of sodium salso of 7-l- [3- (2> 4,6-triinethyl) · plienylicoxasol-5-3 ^r l j-aoetaraido} -oephalosporanic acid

SH / ^S \

N C-CII ₀ -C-N-CH-CH CII ₉

"CH ₅ ^ MK ^ά (j 1 ^ 0

0 = c N , C-CH ₀ -OC-CH ₇

2.8 ml (20 mmol) of triethylamine were added dropwise to a stirred suspension of 2.7 g (10 mmol) of 7-aminocephalosporanic acid in 4-0 ml of dry dichloromevhan at ⁰ ° C. Then 2.55 ml (20 mmol) of trimethylcMorsilane were added dropwise. After the addition had ended, the reaction mixture was ^{kept at 0} ° C. for a few minutes, after which the ice bath was removed. The mixture was then stirred at room temperature for 1 hour. Subsequently, 1.2 ml (10 mmol) of quinoline was added, followed by the dropwise addition of a Lo-about 10 mmol of 3- (2,4 _f 6-trimethyl) -phenyliDOxazol-5-yl-acetyl chloride in 20 ml of dry Diehlormcthan at 5 ⁰ C was added. After stirring for a few minutes at room temperature, the reaction mixture was poured into ice-cold water, whereupon dilute sodium hydroxide solution was added. At a pI value of 7, the layers were separated and the layer was extracted twice with diethyl ether. The organic layers were discarded, and the content was repeatedly extracted with methyl ether at a pH of 5 to 1. The organic layers were

209820/1159 BADORJGWÄL

examined separately thin-layer chromatography · The purest extracts were combined, washed with Waosex ·, dried over anhydrous magnesium sulfate, filtered, concentrated and finally treated with a solution of HalriunH-ethylcaproate in ether. The solid precipitate was collected by filtration, washed with diethyl ether and dried in vacuo to a weight constant, yield 2.5 p. To produce dec Ια * is ta. 13 lionohydrafcij inen the crude product, which contained no other sulfur-containing substances according to the DünncohJchtoirromatographie, recrystallized from acetone · Dan final product (lg) was DVR with the exception of the presence of a small trace of acetone pure, E: · containing 1 Hol V / ater per mole Cephalosporin.

The analysis of the liMR spectrum of the end product, solved in Hexadeutarodimethylsulfoxide (60 Mc, cf values in ppm, internal Standard 2,2-dimethylsilapentane ~ 5-sulfonate) the following values:

NH S _f 26 and 9.12 (Düblect. J ^ O, 5 cps, about 0.8

Protons)

GAl ₉ 6.93 (slightly / widened single toilet,

* 2 protons)

Isoxazolyl-C.-H 6.33 (singlet, t proton) C ₇ -H 5.66, 5.58, 5.52 and 5.44

(weakly broadened signals, * J ^ ö, 5 cpß and J ** A »Ί cps, 1 proton)

Qc ~ tt 5.04 and 4.96 (J. *, «4, T ops)

⁰ ^ V 3 per te

0-CH ₂ 5.20, 4.99, 4.92 and 4.71 (J ^ 12.5 cps}) new

g 3.92 (broadened singlet, 2 protons)

S-CH ₂ ^ 3.72 »-3.43, -3.33 and ^ 5 # 04

(broadened signals »AB-Quartet, 2 prot <men)

P-CH ₃ 2.27 (3 protons)

^. 07 (singlet) I _g 2.01 (singlet) J

209820/111 ^

Example 8

Production of 7 - {- * C3- (2,6-Diehlor) -phenyl-4-raethylieoxa-

-C-N-CH-CH

COOH

A solution of about 1 mmole of trimethyloilyl-7-isocyanate-deacetoxycephalosporanate in 2 ml of toluene was 286 mg (1 mmol) of 3 ~ (2,6-dichloro) -phenyl-4-methylisoxazol-5-ylacetic acid, which was partially dissolved in 10 ml of dry toluene, given. The addition of about 0.1 millimole of 1-isopropylbenzimidazole as a catalyst started the slow reaction (which took about 24 hours at room temperature), When no development of carbon dioxide was rapidly detectable, a stream of dry nitrogen was passed over the surface of the stirred reaction mixture and the contents the reaction vessel was placed in a well-stirred mixture of egg water and diethyl ether. The pH was brought to a value of 6.8, the layers were separated and the fiber layer was twice with diethyl ether extracted. The combined organic extracts were washed twice with ice water. The organic layer was discarded and the combined V / ano layers (70 ml) at a pH of 2.3 with 80 ml of a 2: 1 mixture of

209820/1159

Extracted diethyl ether and ethyl acetate. This extract was washed twice with 5 ml of ice water over anhydrous Magnesium sulfate dried, filtered and completely evaporated in vacuo. The residue, a pale yellow oil, solidified on stirring with dry diethyl ether. The ether was decanted and the plague again twice mixed with ether. The almost colorless plague was in the Vacuum dried to constant weight. Yield 290 mg. The final product was examined by thin layer chromatography, which revealed the presence of only one sulfur-containing compound. The specified structure was confirmed by IE and NMR spectra. The NMR spectrum showed that the final product was about 82% pure, since it was composed of 1 mole of acetic acid and 2.5 moles of diethyl ether (probably crystal bound) per 5 moles of the desired product. The partial analysis of the IR spectrum of the Final product (in chloroform, fourth in cm "*) gave the following:

ί 3500 OH carboxyl ί 3300 NH 1772 C = Oβ-lactam

ί 1730 C = O carboxyl

1690 C = O amide 1380-1430 isoxazole ring absorptions.

BADORiGINAL example

Preparation of the cyclohexylarain salt of 6- -Ca -chloro- [3- (2,4,6-trimethyl) -phonylisoxazol-5-yl] -ac1; amidoj -penicillanic acid

CH _s

C-CH-CN-CH-CH C (CH,) ₉

ci I; You

o = c -ν c

^ COOII. H _o W- (H

In 25 minutes, a solution of 700 mg of trimeth.ylsilyl-6-isocyanato-penicillanate in 10 ml of dry Mehlorinethane was added dropwise to a solution of 700 mg of 1-chloro-1- [3 ~ (2,4,6 ~ trimothyl) -phenylisoxai3ol -5-yl] acetic acid and about 0.03 ml of N-vinylimidazole were added as a catalyst in 25 ml of dry dichloromethane. The reaction mixture was additionally stirred for a further 4 hours. The in situ hydrolysis of the ßilylestero was carried out by adding about 0.2 ml of ethanol at ⁰ ° C. The reaction solution was poured into a well-stirred mixture of diethyl ether and ice cream buffered to a pH of 7. After the layers have been separated, the water layer is extracted once with diethyl ether and then acidified to a pII value of 3.5. The desired compound was incompletely separated from the water layer by extracting it twice with diethyl ether. The extracts were combined, washed with a small amount of single water, dried over anhydrous magnesium sulfate and evaporated in vacuo. The remaining oil was dissolved in 5 ml of acetone. A diluted lo-

209820/1159

BATH

sung by Cyclohexylftmin in diethyl ether vraröe lsmgaaw zubegeben until no further Wiederschlagsbilöung my * or £ o icle The colorless precipitate vrarde by. Filtration genatim ο it, washed with cold diethyl ether and dried in vacuo. Yield 250 mg _e Me. Identity of the final product with the formula given above was confirmed by BKR and Ift ~ Spcktron (KBr disk, 1775 cm " ¹ : C = Oβ-lactam, 1680 ciq" ¹ : C = O amide, 1390 and 1450 em "* ¹ : Iooxasol ring) · The purity of the end product was reduced to about 85 #.

Example 10

Preparation of the salt of 6-i- [3- (2,6-dichloro) ~ hl ^ thlilSl] -acetamidoi-penicillanic acid

Sh ^ ^b \

° " ^H " DD

H C

COOH'i

In the main in accordance with the method described in Example 2, 286 mg (1 mmol) of 3- (2,6-dicloro) «phcnyl-4-methylisoxasol-5-yl-acetic acid kit were 314 mg (1 wKo3) of trimethylsilyl-6 -ioooyanatopenieillanat in 10 ml of dry toluene, in the presence of about 0.01 Kl U-Isopropy'Jbenüijflidar. implemented as a catalyst, Me BeaktiomnniaeJiunc \ / nrdo uhc to & i about 15 ⁰ G g © riüirb. According to the IKinnschichteU.ro ι Lnwaif 4a " Xnmfwa $ £ zvtm miiidoijtoriö 'Ib / ji" last

20M20 / 1159

Produlrt been convicted. The reaction product was treated in the usual manner. At a pH of 3.8, the penicillin was removed from the water layer by / 40 ml extractions with diethyl ether. The combined extracts are washed with ice water, dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo to a volume of about 5 ml. The addition of a solution of sodium a-ethylcaproate in diethyl ether resulted in a colorless precipitate which was collected by filtration, washed with cold diethyl ether and dried in vacuo until it was constant. Finally the product was triturated in a small amount of cold dry acetone. Yield 300 mg. Examination of the end product by thin-layer chromatography, by IR spectrum and. \ MR spectrum confirmed the stated structure. The product was only contaminated with very small amounts of acetone and sodium a-ethyl caproate.

Analysis of the NMR spectrum of the final product dissolved in Ilexadeuterodimethylsulfoxyd (60 Mc, cf-Y / erte in ppm, internal Standard 2,2-dimethylsilapentane-5-sulfonut) the following values:

C ₃ - (CH-) ₂ 1.54 and 1.64 (6 protons)

looxazolyl O ₄ -CH ₅ 1.82 (3 protons)

CH ₂ -CO- at 3? 95 (broadened singlet,

3 protons)

C ₂ -H 4.03 (3 protons)

C ₅ -H and C ₆ -H at 5.5 (multiplet, 2 protons)

C ^ -IU at 7.6 (sharp close splitting

pattern, 3 protons)

Ii-H at 9.3 (doublet, 0.9 protons).

209820/1159

BADOPWQfNAL

of the Cyolohejcylaiainalzes der 6 ~ ö-.liethyl-f 5- (2 ' _f 4 »6-

⁰ H

CIMMI-GH- * I

CJH

we are borrowed according to the expiry described in Example 9 «

ä ^ uimolar © Hengen friaietliylßilyl-e penieillanat vm $ 1 »Methyl-1- [3- (2,4 _f 6-trimetliyl) ^

in the presence of a minor one Ais LögungsinitteX v «ar4 @ dry üiohloraoiliaii uses * Pie llBjwandlung%? ar nacji 6 «hours Stirring at room temperature completed the reaction mixture was treated in the usual way Penicillin extracted with diethyl ether at a "pIfcWe? I" of 4 and finally obtained as the cyclehexylamine salt. the

, the IE spectrum (intensive β-lactaacarbonyl absorption at 1778 cbi "" ¹ (KBr-Seheibe)) and the HI> 3R spectrum confirmed the stated structure of the end product and the good purity of the product,

The feiXanalye © of the complicated © «COW spectrum of the. End product ^ O {© ine mixture of D- and dea L-isoiaeren), in Hexadeutei? © diBiethylsulfoxyd (60 Hc, <f-values in ppm, in

209820/1159

external standard 2,2-dimethylsilapentane-5-sulfonate) gave the following values:

N-H

C ₆ H ₂

Isoxazolyl-C.-H

Cc-H and 5

C _a -HC ₂ -H

Cyclohexyl-Cj-II

T) -CIL

Cyclohexyl C ₅ II C ₅ - (CH ₃ ) ₂ and C ₀₁ -CH ₃

8.9 (about 1 proton)

6.95 (etv / as broadened singlet, 2 protons)

6.35 (2 narrow singlets, 1 proton) at 5.4 (2 protons)

at 4.2 (diffuse quartet) at 4.0 (2 narrow singlets)

_{2 protons}

at 2.9 (broad absorption range, about 1 proton)

2.3 (singlet, about 3 protons) 2.1 (singlet, about G protons) at 0.9 - > at 2.3

approx.

26 pro tones

at 1.4

at 1.65

209820/1159

Example 12

Production of the sodium sala of the 7 ~ £ ~ [? - ('', 4 » phenyl-4-mothyliGOxazole -!> ~ ylJ-rtcOiP luloj-oc < , ^

H C- CH ₀ - C - Ii - CH- (JH CH _{0 n}

CH ₅ ^ -0 ^ ² II | ² "

coo; »» ·.

1.38 ml (10 bjMoI) TriethylaI in were added dropwise -to a stirred suspension of 1.3 g (5 rallol) 7-AL'iinocephalorjporansäijre in 20 ml teockeneia DichloiViu than cei 0 ° 0. Subsequently, 1.26 ml (10 inMol) Triaie I ¹ ^ yIo iT. Oil were added dropwise at ⁰ ° C. After the end, the addition of ¹ % of methyl chloroilane was added to the reaction mixture for a few minutes ^{Stirred 0} ° C., before the ice bath was removed. Then during 1 stutlciö-bei'RauxffcenperaUir gox ^ ührt. Then 0.6 ml (5 mmol) of quinoline were added, whereupon a solution of ctv / a 4, ^f -> w! 1ol

chloride (about 90 igor fineness of 1.3 C (5 milol) of the corresponding carboxylic acid) was added in 10 nl of dry dichloromethane at 5 ° C., and the reaction was quenched for one minute with stirring at room temperature fermented in Eiowactcr; r, on. I ¹ Sr pU-V / cvt vurd «to 7 t-rhoht and the layers were separated. DLe VafjHerijehu-Ut, the geiaaf3 of the thin-oliichtchroaiatographiochen investigation an initial reaction product (a small amount of 7-amino ~

209820/1159

** JT3 > w

""egg

oa) contained, wrdo pweljaal with BlätlyrJäihey Pie oi ^ anieclion 8p | 4.cliten was «vervoyfeiu The poljicht wniHie, a» oehj.ioß <* «ä at pli - \ / eyten van 5, O _f 4 * 5 4 | 0 »It dietljyl ^; nhey exlrafelevt «Bor Ji ^ irakt at pH ^ ntliioli lecU ^ liaJi d« a gex / ünscJite lia \ iftpyo4ukt _t The pinei 'liöqune of liaty4wa- ^ iitJiyl0aj> r «Bat PW dle yielded c; ine« colorloee ^ foptem Yield 1.2% by weight dor JKinmscfracfciuhyoiaatogj ^ jihiQ »dop JR-Spekiren was üa, ® Mnd ^ oaxkki. lediglieli pit ¹ 3 '. «τΛνι (ulwa 1

DIo analysis äea '\ IIlß-Sppkti. * Ai. »R ^

piiv-r about 2} 1-kjschu »g of loading enterodiiaetliyleulfoxy4 v "nd 1) ^ 0 (60 Hc, cf-words in ppn, internal standard

the

G ₆ H €, 95 (SinguXott, 2 protons)

C ₇ -H b _f 12 \ ma 5.64 (doublet, J = ^ 4.6 epq, 1

C ₆ -H 5.10 wnd $, 02 (j) ubl <3tt _# J «<4» 6 c

bBi 5.15, 4.92, 4.81 and 4.59 (AB-Q \ jartctt, J-13.2 pps)

at 5.55 (Zenirwi »© in ^ js A ^ -Qu ^ rtetts) /. 3 * 87 (ctvas broadened singlet) j tone

₃ 2.28 (3 protons)

0-CO-CH ₅ 2.05 (sinculet) 7 ₉

(o-CH ₅ ) ₂ 1.98 (singlet) j

Iijoxaaoly 1-C ₄ -CH ₇ 1.71 (3! Photons).

209820 / 11S9

Example 13

Preparation of the sodium salt of 7-i ~ [3- (2,4,6-trimethyl).

phenyl-4-methylisoxazol-5-yl] - acetamidoJ ~ desacetoxy-

cephalosporanic acid

oh ^

-cn- ch ch "ch ₀

Essentially according to the procedure described in Example 9, 1.3 g (5 mmol) of 3- (2,4,6-T: .'imethyl) -phenyl-4-methylisoxazol-5-yl-acetic acid, dissolved in 25 ml of dry dichloromethane and 5.04 mmol of trimethylolyl-7-ioocyanatodesacetoxycephalosporanate, dissolved in 9 ral toluene in the presence of about 0.05 ml of N-vinylimidazole reacted as a catalyst The addition of the isocyanate in toluene took place in about 20 minutes. The evolution of carbon dioxide was already after 5 minutes to determine. After about 7.5 hours of additional stirring, the reaction was interrupted as a thin layer chromatogram of the reaction mixture shows an approximately 80 conversion of isocyanate into the desired product and the evolution of carbon dioxide had almost ceased. The reaction mixture was then treated in the usual manner. The cephalosporin was at a pH of 4.5 with the aid of a 9il foi3chuug of diethyl ether and ethyl acetate extracted. The combined extracts were washed with mild water, over anhydrous magnesium sulfate ßefc

209820/1159

net, filtered and completely evaporated in vacuo. The remaining oil was dissolved in diethyl ether. The addition part of the estimated required amount of sodium a-ethyl caproate, dissolved in diethyl ether, led to a precipitate, which was collected by filtration with washed a small amount of cold diethyl ether and dried in vacuo. The combined filtrates were Sodium cc-ethyl caproate was added again. The received second batch of solid material was treated as the first batch. The third and final batch was received by adding dissolved sodium ct-ethylcaproate, up to there was no further increase in the FieC blow. the The third batch, which was practically pure according to thin-layer chromatography, was recrystallized from acetone. In the end the three batches were redissolved in acetone. the Acetone solution was slightly concentrated in vacuo and then inoculated. After crystallization, the flask was transferred to the freezer. The next day they were Crystals collected by filtration, mxt cold acetone and Diäbhyläther washed and dried in vacuo to constant weight. Yield 1.7g. The specified structure was confirmed by IR and NMR spectra NMR spectrum and thin layer chromatograms was the final product only by a very small amount of acetone and a small amount of NjN'-didesacetoxycephalosporanic acid urea contaminated.

Analysis of the IIMR spectrum of the end product, dissolved in an approximately 2: 1 mixture of ilexadeuterodimethyl sulfoxide and D ₂ O (60 Mo, cT values in ppm, internal standard 2,2-dimethylnilapentane-5-sulfonate) gave the following:

C ₆ H ₂ 6, SY (slightly broadened singlet,

2 protons)

209820/1189

BADORiQtMAL

C ₇ -H and C ₆ -H 5.63, 5.46, 4.97 and 4.90

(AB quartet, J * «4.5 cps, 2 protons)

CH ₂ -CO 3.86 (broadened singlet, 2 protons)

S-CH ₂ about 3.7 → 2.9

(AB quartet, <Τ? «17 · 5-1 cps, 2 protons)

P-CH ₃ 2.29 (3 protons)

(o-CH ₃ ) ₂ 1.98 (singlet) 7 _{9 Prot (m <m}

C ₃ -CH ₃ 1.94 (singlet) J.

IsOXaZOl-C ₄ -CH ₃ 1.71 (3 protons).

Example Η

Preparation of the 6 - {- [3- (2,6-dichloro) -phe.iyl-4-carb? Nylisoxazol-5-ylj-acetamidoj -penicillanic acid

C - CH ₂ - C - N - CH-CH

O = CN ^C -COOH

600 mg (2 mmol) 3- (2,6-dichloro) -phenyl-4-carbamyl-3soxazol-5-yl-acetic acid and 63O mg (2 mmol) of triniethylsilyl-6-icocyanofco · penioillanat were dry in a mixture of 15 ml Benzonitrile and 15 ml of dry tetrahydrolurane dissolved, whereupon about 0.02 ml of II-Melhylimidnaol was added directly. The development of carbon dioxide increased after 3 hours of listening Room temperature drops sharply. A thin-layer chromatography of the Reaction mixture showed good isocyanate conversion

209820/1159 BATH ORIGINAL.

at. The reaction mixture was poured into a well-stirred, ice-cold mixture of 30 ml v / ater, 20 ml diethyl ether and 20 ml ethyl acetate. Then dilute NaOH solution was added to pH 8.5. The layers were separated and the water layer was purified by extraction with diethyl ether. The organic layers were discarded and the water layer was washed at pH 3.0 with a 1: 1 mixture of diethyl ether and ethyl acetate. The combined extracts were washed with ice water, dried over anhydrous magnesium sulfate, filtered and evaporated completely in vacuo. The resulting pale yellow solid + rff (1.1 g) was examined by IR and NMR spectra. The product contained the desired penicillin but also small) amounts NtH'-Dipenicillansäurefcarnstoff and Ausgangbcarbonsäure. To prepare a more accurate sample, the crude product was extracted repeatedly with cold, dry diethyl ether, in which the urea is hardly soluble. The ether extract was mixed with ice water, which was buffered to a pH of 7. The greater part of the starting carboxylic acid was removed from the water layer _{at a pH value of 4 f 5} and extracted after increasing amounts of ethyl acetate. The extracts, which were free from the starting carboxylic acid, the urea and the degradation products, were combined and completely evaporated in vacuo after the usual manipulations * The resulting colorless solid was dried to constant gray weight (350 mg). According to the thin layer chromatograms, the IH and the HMR spectra, the end product was pure except for the presence of small amounts of ethyl acetate and diethyl ether. The IR spectrum (KBr disk), complicated by the presence of the monomers and the dimers, showed a broad, intense range between 3000 and 3600 cm "" ¹ with band

209820/1159 BADORKSfNAL

at 3450ι 3350 and 3200 cnT, the NH absorptions both amide groups are attributable to carboxy OII absorption at about 2550 cm "", a broad, very intense carbony! .absorption with bands at + 1790, ± 1725, + 1

+ 1695 and Z 1655 cm, which are to be assigned to the ß-lactarn, the carboxyl group, the CO'IJH group or the C0 »NH ₂ group.

Example 15

Preparation of the sodium salt of 6-t- [3- (2,6 -]) dichloro) -phenyl-4-cyanoiso; azol-5-yl] -acetamido] -penicillanic acid

C = N

{ 2 H

C - CH ₀ - C - N-CH-CH

O = C N C

^N COONa

297 mg (1 mllol) 3- (2,6-dichloro) -phenyl-4-cyano-jooxazol-5-yl-acetic acid, 3H mg (1 mmol) trimothylsilyl-6-isocyunatoponicillanate and a trace of H-isopropyl alcohol dissolved in 5 ml Trockonom dichloromethone. According to dcu J) ünnschichtcbrouatograuiTi) was a good inocyanate conversion after 3-hour reaction at room temperature reached, Dau R (, - a] {- ionization product It was treated in the usual way. Polish it was elin solution of the penicillin in water dir / di J.xtraktioneii irijt Diüthy la liier at a pH value of 7.0 and 4.5 gcrojnigl. ]) as penicillin was »by extraction ladt Piiitli.vl-

209820/1159 BAO 0RK3INAL

ether removed from the water at a pH of 3.3. The ether extract was washed with ice water, dried over anhydrous magnesium sulfate, filtered and evaporated in vacuo. The remaining oil was dissolved in about 3 ml of dry ethyl acetate, whereupon about 0.6 mmol of sodium ct-ethyl caproate, dissolved in a small amount of ethyl acetate, was added · The addition of dry diethyl ether resulted in a pale colored precipitate which was collected by filtration, washed with cold ethyl ether and dried in vacuo to constant weight · Yield 180 mg. The end product was examined in the usual way · The sodium salt contained the desired penicillins and small amounts of a degradation product and sodium d-ethyl caproate · The IR spectrum of the end product »(KBr disk) showed, among other things, absorptions at 2260 (C ^ N), 1778 ( Carbonyl-ß-laotam), 1690 (carbonylamide), 1610 (carbonyl carboxylation) and - 14-00 cm (isoxazole ring absorptions).

209820/1159

Example 16

Preparation of the sodium salt of 6-f- [3- (1) -ada »antylisoxaeol-5-yl J-acetaBidl -penieillanpäure

C - CH ₉ - C - H - CH-CH

Ib veoeTi tical according to the procedure described in Example 9 became a solution of 780 mg (2.5 mmol) of trimethylsilyl-6-isocyanato-penieillanate in 10 ml of dry dichloromethane dropwise to a solution of 650 mg (2.5 mmol) 3- (1) -adamantyl-isoxazol-5-yl-acetic acid and about 0.02 ml of N-vinylimidazo? in 20 al dry dichloromethane given. The conversion was after a total of 2.5 hours Stirring ended at room temperature, which was indicated by a drastic reduction in the evolution of carbon dioxide. A thin layer chromatogram indicated good conversion of the isocyanate to the desired penicillin. The reaction product was treated in the usual way. The penicillin was isolated by double extractions with diethyl ether, which is carried out at pH 5 »5 or pH 4.0 were extracted from water. The extracts were separated, washed with ice-cold water, over anhydrous magnesium sulfate dried, filtered and evaporated completely in vacuo. Yields 700 and 300 mg, respectively. Both products yielded satisfactory IR spectra and contained in accordance with thin film chromatography only contains a penicillin. There

209820/1159

the sample obtained by extraction at a pH of 5 | 5 was obtained, contaminated by the Auagangsessi & isäurederivat it was dissolved in ether, whereupon sodium ethyl caproate was added. The obtained sodium salt (350 mg) was except for a small amount of Pure sodium ci-ethyl caproate. According to the NMR spectrum, the second product is pure with the exception of a small amount of diethyl ether (about 4.0% by weight).

Partial analysis of the IR spectrum of the sodium salt of the final product (KBr-Schelbe, Verte in cm ") resulted in the following»

1605 C = O carboxylate ion

- 1520 NH-Pofo'ination oscillation ± 1405 isoxazoleing absorption

- MOO NH 2910
2853 CHp groups 1775 C = Oβ-lactam ί 1675 C-O amide.

The anu'yoe of the HMR spectrum of the end product (the acid), dissolved in llexadeuterodimethyl sulfoxide (60 Mc, </ * ~ VJerte in ppm, internal standard 2,2-dimethylsilapentane-5-sulfonate) the following:

H-II at 8.9 (about 0.8 protons)

ΙΓιλ 1 1 * "3 Γ7 f \ 1" IT ι C ^ m * TT r \ ^ r \ 1 1 "15-w f \ -4 ~ f \ > Λ Λ

ij Ki si ei Zr U. I JJ --Ό α ~ · .Π O | flO ^ \ XlUuUiiy

C ₁₃ -JI and C ₆ -H 5.35 - i> 5.60 (Multiplelt, 2 protons)

C ,, - II 4.26 (1 proton)

Tf-ClI ₂ 3.79 (broadened sin ^ ulet, 2 protonons)

Aaam <"ni ^ l _L r- <<ppe 1.90 and 1.73 ->)

Zcntri. a ο l \ iuo \ (rbrr-i 1- ^ ', er · <Ab »or]) Lions)?" ca,

0, - (CH Ji. _? 1.64 xuA 1.51 J ^{? 2 Px} '°

209820 / 11S9 BADORJQiNAL

Example 17

Preparation of 6- $ t- (p-nitro) -benzyloxycarbonylamino- [3- (2,6-dichloro) -phenylisoxazol-5-yl! -Acetamido] -petiicillana

acid

-NH- CH-CH C (CH,) _o

COOH

-KO,

2.33 g (5 mmol) 1- (p-nitro) -benzyloxycarbonylamino ~ 1- [3- (2,6-dichloro) -phenylisoxazol-5-yl! -Acetic acid, 1.57 g (5 mmol) trimethylsilyl- 6-isooyanato-penicillanate and 0.1 ml of N-vinylimidazole as a catalyst were dissolved in 50 ml of dry dichloromethane. After stirring for 3 hours under nitrogen at room temperature, the conversion was complete. According to thin-sheet chromatography, about $ 70 of the isocyanate had been converted to the desired product. The reaction product was ^{cooled to 0} ° C., after which a few milliliters of cold acetone, which contained enough water to hydrolyze the silyl ester, were sucked in.

puts. The mixture was then completely evaporated in the cold in vacuo. The residue was dissolved in 75 ml of a cold 1ί1 mixture of diethyl ether and ethyl acetate. Since this penicillin was to be used for the production of the penicillin of Example 18, the isolation

208820/1159

BAD ORIQtNAL

longitudinal process is not carried out to isolate the product in an essentially pure state but to isolate as large an amount of the product as possible. For this purpose, the solution was mixed with 70 ml of ice water that was buffered to a pH of 7. The well-stirred mixture was acidified to a pH of 5.8 and transferred to a separatory funnel. The water layer was drained and discarded as it contained the by-product N, N'-dipenicillanate and only traces of the desired product. The organic layer was then washed twice with weakly acidic ice-cold water and once with a small amount of neutral water a. The organic layer, which had been completely freed from the urea and the catalyst in this way, was dried over anhydrous magnesium sulfate, filtered and completely evaporated in the cold. The residue was dried to constant weight in vacuo. Yield 3.4 g of a pale yellow, essentially crystalline solid. The thin layer chromatograms of the crude end product indicated the presence of only the desired penicillin and the starting protected amino acid in a ratio of about 2: 1. This was confirmed by the WMR spectrum, which also indicated the presence of ethyl acetate and a small amount of water. The calculated content of the desired penicillin in the crude product was about 2.2 to 2.4 g

209820/1159

Example 18

Preparation of 6- £ ^tt ~ amino [3- (2,6-dichloro) -phenylisoxazol- 5-yl! Acetamido] "-penicillansaure

H _x

-N-CH-CH

O = C N

C (CH ₃ ) ₂

3.0 g of the crude product of Example 17 which contained about 2 g of 6 ~ {a- (p-nitro) -benzyloxycarbonylamino- [3- (2,6-dichloro) -phenylisoxazol-5-yl! -Acetamido] -penicillai : Containing 3 acid were dissolved in 100 ml of ethyl acetate, mixed with 25 ml of water. The pH of the mixture was brought to a value of 7.0 by adding a dilute sodium hyuroxide solution. After 1.5 g of Pd / C 10 ⁰ Jo had been added , hydrogen was continuously passed under the surface. Analysis by thin-layer chromatography revealed that the reaction had ended after stirring for 135 minutes at room temperature. Nitrogen was bubbled through the reaction mixture for 10 minutes, then ice water was added and the pH was brought to 4.7. The contents of the funnel were transferred to a separatory funnel. The mixture separated into a clear ethyl acetate layer and a water layer separated by an emulsion layer. The water layer was drained and stored. Then the emulsion layer was centrifuged. The resulting layers were go-

209820/1159

BADORFOLNAL

separates. The ethyl acetate layer was pre-cleaned with the first ethyl acetate layer. The water layers were also combined and extracted again with ethyl acetate. The water layer was then discarded. The remaining catalyst was removed from the combined ethyl acetate extracts by filtration. The colored filtrate was ^{concentrated at 0} ° C. in vacuo to a volume of about 25 ml. Then 100 ml of ice water were added and the mixture was brought to a pH of 7 * 0. The layers were separated and the colored organic layer was discarded. The solution of the desired compound in water was extracted twice with a 1: 1 mixture of ethyl acetate and diethyl ether. The resulting practically colorless solution in water was acidified to a pH of 4.7 and extracted twice with an excess volume of ethyl acetate. The water layer was discarded and the combined ethyl acetate layers were washed twice with a small amount of ice water. The thin-layer chromatograms of the final extract showed an extensive (the mixture is a D, II mixture) sulfur- and ninhydrin-positive spots. After complete evaporation of the extract, a slightly colored pesticide was obtained. Yield 650 mg of dry material. The end product was investigated with the help of the IR and NMR spectra and was in the form of a 1: 1 molar mixture of the desired penicillin and ethyl acetate, possibly with small amounts of a 3- (2,6-dichloro) phenylisoxazole derivative is contaminated.

The partial analysis of the IR spectrum of the end product (KBr disk, Values in era "") resulted in the following values:

at 3300 ITH Carboxyl at 2600 OH ß-lactam 1780 C = O

20i82G / 11S9

ί 1750 C = O ethyl acetate

- 1705 C = * 0 carboxyl 1690 C = O amide 1590 υηα 1440 Isoxazole ring absorptions 785 C-Cl.

Example 19

Preparation of the sodium salt of 6- £ - [5- (4-Hitro) -phenylisoxazol-5-yl] -acetafflido-penicillin acid

CH _Λ

0 H

- CH ₀ - C - N - CH - CH C (CH,) _O

<- Il I J * -

O = C N -, C

^ OOONa

In the usual way, 166 mg (0.67 mmol) of 5- (4-nitro) -phenylisoxazol-5-yl-esoetic acid, 210 nitf (0.67 mmol) of trimethylsilyl-6-isocyanato-penicillanate and a trace of N-isopropylbenimidazole were used . 5 ml of benzonitrile were used as the solvent. The reaction was complete after stirring for 5 hours at room temperature. The contents of the flask were poured into an ice-cold, well-stirred mixture of 25 ml of water, 20 ml of diethyl ether and 25 ml of ethyl acetate. By adding a dilute HaOH solution, the acidic mixture (pH 5) was neutralized to 3 in pH-value of 7 and the layers were separated. The organic layer was verw # RFEN and WassorBchicht was for cleaning once with

209820/1159

BAP ORlGlNAi

40 ml of a 1: 1 mixture of ether and ethyl acetate was extracted. Then 30 ml of a 1: 1 mixture of ether and ethyl acetate were mixed with the water layer and the pH was lowered to a value of 3.5. The layers were separated and the inactive layer was re-extracted with 50 ml of the same mixture of solvents. The combined organic extracts were washed twice with a small volume of ice water, then dried over anhydrous magnesium sulfate, filtered and evaporated completely in vacuo. The resulting yellowish oil was triturated with dry diethyl ether. The partially crystalline contaminant formed was collected by filtration and then repeatedly stirred up in ether. After drying in vacuo, the colorless product ultimately obtained weighed 73 mg. Examination of the product by thin-layer chromatography and by NMR spectrum showed that the desired penicillanic acid contained 5 to 6 moles of water per mole of the compound and a small amount of diethyl ether, but was otherwise pure. The ether filtrate obtained and the ash solutions were completely evaporated. The residue was dissolved in 3 ml of a dry 1: 1 mixture of ether and ethyl acetate and then treated in the cold with a dilute solution of sodium a-ethyl caproate in ether. The precipitated sodium salt of the desired penicillin was collected by filtration and washed repeatedly with dry ether. After drying, this product weighed 134 mg. The product was tested in a conventional manner · Apart from adhering water (far less than is au in the free Ponicillansäui'e the FmII) was the purity of liatriiimsalzies to about PQ'm 85 $ appreciated since on about 5 Gev .- # of Abbauprodukteα and 10 to 15% by weight of jiatrium a-ethyl caproate contained.

The analysis of the «liMR-üpektruitte the in a mixture of about 6 parts of Hexadeuterodimethyltsulf # xyd and 1 part of DgO dissolved

209820/1159

6 - [- [3- (4-Nitro) -phenylisoxazol-5-yl] -acetamido} -penicillanic acid (6O Mc _f <f-Y7erte in ppm, internal standard 2,2-dimeth / isilapentane-5-sulfonate) the following values:

C ₆ H. 7.95 - ► 8.4 (AA ¹ BB ¹ splitting pattern,

* 4 protons)

Isoxazolyl C.-H 6.94 (1 proton)

Cc-H and C ₆ -H 5.5 (slightly broadened singlet ³ , 2 protons)

C ₂ -H 4.15 (1 proton)

4.0 (slightly broadened singlet, 2 protons)

₅₅ J ₂ 1.63 and 1.52 (6 protons),

IHo partial analysis of the IR spectrum of the liatriuasalsos of 6- {~ [5- (A-KitroJ-phenylisoxazole - ^ - ylJ-acetaiaidoJ-penicillanic acid (KBr ~ disk, values in cm ") resulted in the following values:

at 3380 HH

1770 C- = 0 J? -Lictan »(intensive)

1675 C = O amide (intense)

1600 C «0 carboxylation + probably aroraati-

see α C ^ C (very intense)

1525 HO ₂ + raögl. HH Def. (Oohr intensive)

1400 - 1460 leoxazole ring absorption 1355 HO ₂ (intensive)

ί 858 C-MOo «nd aroiaatiöclie«

* (medium intensity).

209 · 20/1159

BAD ORIGHiAL

Example 20

Preparation of 6-t <* ~ carbamyl- [3- (2,6-alehior) -plienylisoxazol-5-yl! -Acetamido! -Penicillic acid

GH 0 _H 1 Bv C-CH-CN-CH-CH

O. O = C N C

To a solution of 1.0 g (3.7 mmol) of 3- (2,6-dichloro) -phenylisoxazol-5-yl-acetamide in 15 ml of dry tetrahydrofuran which had been cooled to -7O ⁰ C were added dropwise a solution of 3.7 mmol of n-butyllithium in hexane. The addition \ ru \ ie so adjusted that the reaction mixture was maintained below -60 ⁰ C. After a few minutes further stirring at -70 ⁰ C 0.47 ml was added dropwise (about 3.7 mmol) of freshly distilled trimethylchlorosilane was added. Then the cooling bath was removed and the temperature allowed to rise to -30 ⁰ C. This procedure - the successive addition of 1 equivalent of n-butyllithium and 1 equivalent of trimethylchlorosilane - was repeated in equal amounts. _{0.56 ml (3.7 mmol) of N 1} N, N were added to the solution of the Ιί, Ν-BiG-triraethylsilyl derivative of the Auegangsprodukteu in a mixture of 15 ml of tetrahydrofuran and about 3.5 ml of hexane, prepared in situ in this way «, II · - Tetramethyliithylcndiamine. A solution of about 3.7 mmol of ii-butyllithiura in 1.76 ml of hexane was added dropwise to the mixture, which had been cooled again to -7!> ° C U>. The addition

209820/1159 BAO OR (QlIiAI.

- 5 »-

The speed was set in such a way that the reaction temperature was a maximum of −0 ° C. The reaction mixture was finally stirred for 1 hour at -70 to -6O ⁰ C. The reaction sequence was duroh the dropwise addition of a solution of 1.16 g (3.7 mmol) of trimethylsilyl-6-isocyanatopenicillanat in 10 ml of dry toluene, wherein the reaction temperature was kept below -55 ⁰ C, terminated. The reaction mixture was stirred additionally during 30 minutes at -60 ⁰ C. The reaction mixture and dilute hydrochloric acid were slowly and simultaneously poured into a well-gertihroa ice-cooled mixture of 50 ml of diethyl ether and 50 ml of water with a pH of 4. The pH-Ve * t of the mixture was then increased to 7 and the layers were separated. The water layer was extracted one more time with 50 ml of ether at pH 7. The organic layers were discarded. The water layer was extracted three times with ether at pH values of 5.0, 4.5 and 4.3 and once with a 1 »1 mixture of ethyl acetate and diethyl ether at a pH value of 4.3. By thin-layer chromatographic investigation it was found that the water layer no longer contained the desired penicillin together with small amounts of sulfur-containing impurities and that the first three ether extracts contained the penicillin in an essentially pure state. The ether extracts were combined, washed with ice water, dried over anhydrous magnesium sulfate, filtered and completely evaporated in vacuo. The pesticide obtained weighed 500 mg after prolonged drying in vacuo. The IR and NMR spectra confirmed the stated structure of the penicillin. The protected purity was 80 to 85 #.

The partial analysis of the IR spectrum of the end product (KBr disk, Values in cm "*) resulted in the following values:

209820/1159

- 3440 NH (probably CO-NH ₂ ) - 3330 NH (probably CO-NH) + 3210
mm NH (probably bound NH) 2500-2650 OH (carboxyl) 1780 C = Oβ-lactarn - 1720 C = O (carboxyl) 1690 and 1660 C = O of CO-NH and CO-NH ₂ 1598 aromatic C = C and NH ₂ deformat ί 1525 probably NH deformation 1395, 1430 Isoxazole ring absorptions

790 C-Cl and aromatic substitution pattern

Example 21

Preparation of 6 - {- [3-Methylisoxa4ol-5-yl! -Acetamido} penicillic acid

Λ f HH ^ S GH ₃

N Jf - CH ₉ - C - B - GC O ^

ti

^CH 3

c-ir < ^H

tr ⁿ cooh

Essentially according to the procedure described in Example 2 282 mg (2 mmol) of 3-ethylisoxazol-5-yl-esta-acid were obtained with 628 mg (2 mlol) trimethylsilyl-6 ^ *. iBOcyanatopennicillatiat in 10 ml of dry dichloromethane in the presence

209820 / 11S9

of three drops of II-isopropylbenzimidazole as a catalyst implemented. After the usual work-up of the reaction mixture a pale-colored product was obtained which, according to thin-layer chromatography and the IR and NMR spectra, was obtained had a good purity.

Analysis of the NMR spectrum of the mixed in some DpO Hexadeuterodimethylsulfoxide dissolved product (60 Mc, ef values in ppm, tetramethylsilane as internal standard) the following values:

Isoxazolyl C, -H 6.22 (2 protons) C ₅ -H and C ₆ -H 5.50 C ₂ -H 4.32 (2 protons) CHg-CO- 3.82 Isoxazolyl-CH- 2.23 and 1.52 C, - (CH,) o 1.65

The partial analysis of the IR spectrum of the end product (in KBr, Values in cm **) resulted in the following values j

- 3500 OH carboxyl 3350 NH 17SÖ C = O ß-lactara 1740 C = O carboxyl 1670 C = O amide 1380-1430 Isoxasol ring absorptions

209820/1159

Example 1 22

Preparation of 7- £ - [3-methylisoxazol-5-yl] ~ acetainido} < c ephalosporanic acid

HH ^ \ C - CH ₀ - C - N - C - C CH ₉

ο Il I

N C-CH ₀ -OC-CH,

^ C ^ 0

COOH

Following the procedure described in Example 3, 3-methylisoxazol-5-yl-acetyl chloride (prepared from 4.5 mmol 3-methylisoxazol-5-yl-acetic acid and thionyl chloride) with IT, O-BiB-trimethylßilyl-7-ainino-cephalosporanate ^ made from 1.224 mg (4.5 mmol) 7-aminocephalosporanic acid) implemented

After the reaction and the work-up of the reaction mixture, 790 mg (44 %) of a pale yellow product with a purity according to the thin-layer chromatography of the IR and NMR spectra of about 70 °.

The analysis of the NMR spectrum of the final product, dissolved in a mixture of deuterochloroform and Hexadeuterodimethylsulfoxyd, the bit D ₂ O was added (60 Mc, ^ -Vierte in ppm, tetramethylsilane as an internal standard) _ga · b the following Y / erte:

209820/1159

2t55Q8t

Ißoxazolyl O, -H 6.12

C «-H 5.75 and 5.66 (Js ^ 4.5 cpe, 1 proton)

Cr-H 5.05 and 4.97 (J «i4.5 cpe * 1 proton)

CH ₂ -CO- 3.78 (i 2 pr οtoning ) 2 protons) 0-CH ₂ - 5 * 22 - - * 4 * 68 ( J ^ 13 cpe » 2 protons) S-CH ₂ 3.80 - ■ * 3 »15 ( Yes ^ IB Cp0, CO-CH ₅ 2.07 Isoxazolyl-CH, 2.25

The analysis of the HI spectrum of the end product (KBr, Values in cm "*) resulted in the following:

£ 3,280 OH 1760 CsO ß-lActam 1750 C = so Effter 1670 C = O amide 1230 C-O-C esters 1380 and 1420 Isoxazole Ring Absorptions ^

example

Preparation of the sodium salt of 6-fä- (N-phenyl) -oarbamyl- [3 ~ (2,6-dichloro) -phenylisoxazol ~ 5-yl! -Acetamido! -Penicillanic acid

C-CH-C-

N -. CH-

C = O

.0-

CH

I I

N-

■ AL ^N COONa

A solution of 1.0 g (2.9 mmol) of N- £ henyl-3- (2,6-dichloro) -phenyliaoxaaol-5-yl-acetamide in 15 ml of dry tetrahydrofuran was cooled to -70 ⁰ C. At -70 ⁰ C a precooled solution of about 2.9 mmol of n-butyllithium in 5 ml of a mixture of n-hexane and dry tetrahydrofuran, and then 0.44 ml (about 2.9 mtyol) Ν, ΙΙ, were successively added dropwise. Ν ¹ , H'-tetramethylethylenediamine was added and finally a precooled solution of 2.9 mmol of n-butyllithium in 5 ml of a mixture of n-hexane and dry tetrahydrofuran was added. The Heaktionsraißchung was stirred for 1 additional hour at -70 ^0C. To the wines produced in this reagent were then added dropwise at -70 ⁰ C a Löourg of 0.91 g (2.9 mmol) irimethyl £ jilyl-6-ioocyanato-penicillanafc in 5 ral dry toluene. ! laughing completion of the addition the temperature was allowed the Roaktionomischung to -50 ⁰ C rise and touched the Heaktionamiachung at this temperature

209820/1159 BATH ORIGINAL

about 30 minutes. Mused wurdendie reaction mixture and dilute hydrochloric acid added simultaneously to a solution, cooled to 0 ⁰ C well-stirred mixture of 30 ml WaBser and 30 ml of diethyl ether · The additions were balanced such that throughout the neutralization, a pH value of about 7 »5 ceased · The The resulting layers were separated and the water layer was extracted once with 30 ml of diethyl ether and once with 30 ml of ethyl acetate for purification of diethyl ether and formic acid was used. The combined organic layers containing no such compounds were discarded. The chromatogram of the water layer showed four well-developed spots, three small spots and one major spot.

The smaller blanks were assigned to the degradation products, NjN'-dipenicillanyl urea and n-butylcarbonaraidopenicillanic acid. The Rf values of the latter two picks were equal to that of the real penicillins. In order to separate the compound which was responsible for the fourth and the main spot of the chromatogram, the water layer was extracted with 30 ml of diethyl ether at pH 4.9 and pH 3.6, which led to a complete separation of the desired compound from the water layer. The remaining material and part of the third compound (presumably n-Dutylpcnicillin) were separated by extraction at pH 3-3 with a 2: 1 -Iiiijchung of diethyl ether and ethyl acetate :. In order to remove then TiobenprocJu / Tt au, this layer was v / i ed recovered with EicwaBser with a pH value of 4.6 wanclien, waa % \\ a third, almost pure extract and a number of

209t20 / 1159

BATH ORIGINAL

-63- 2T55Q8?

Wash waters that still contained significant amounts of the desired product resulted. The fourth extract was made through Extraction of the combined wash water with ethyl acetate pH 6.0. The four extracts were combined, washed with ice water, over anhydrous magnesium sulfate dried, filtered and concentrated in vacuo «The concentrated solution of the desired compound in Ethyl acetate was treated with a concentrated solution of sodium c-ethyl caproate in ethyl acetate. That The i? atrium salt of penicillin was precipitated from this solution by adding dry diethyl ether. Schlag uurüe collected by filtration with diethyl ether washed and bic dried to constant weight. Yield 580 mg. The final product was thin-filmchroroatographically and examined with regard to the IR and HMR spectra, whereby the stated structure was confirmed and it was established that the impurities in the end product a small amount of sodium a-ethyl caproate and were a small amount of degradation products

The partial analysis of the IR spectrum of the end product and dea Starting product (dissolved in chloroform, concentration about 10 mg / ml, values in cm) gave the following values

209820/1159

· »64 * ·

2155061

t 3420

t 5500 1775

t 1705

1600

1558

End product

KH
(probably

acetaaid
5450

m (board)

ß-LactaiB (intensive)

0 * 0 AmU

(very intensive)

OwO carbonylation

ar on # (

1695 QmO Äjnifl (intensive) 1596 C »0

0C u / ode 0N (hard, medium Internet)

* 1550 likely HH deformation (medium intensity) (intenγiv)

1557 0 * 0 and / or (echarf _f mtWXem sitat)

ί 1520 vmbreofeeinlioh Iteforwitlone

(middle

0 «0 arosw {bitter intensity)

t 1440
1380

(KBr-Scheibö) C-Ol (intensive) 1496 0 rn * (medium intellect)

arlttlere 1 intensity C ring-

Bitter ones
Intensity

1435 intensive)

( ring-

1380 mean f tiS intensity ) * ^ ^ηβη

785
(KB

«Older

(KBr-ßcheibe) literally aro-

roatißcheo Bubstl- Boheinlioh tutioneseuster

Intensity) (average intensity)

-. 65 -

B e i a pie 1

Preparation of 7- £ - [3- (4-Hitro) -phenylisoxazol-5-yl] -acetamidoul -cephalosporanic acid ______

Il

-CHo-C- NH -

CH-CH

I I

σ - ν

CH

COOH

, -0-C-CH.,

According to the procedure described in Example 3, 3- (4-2 ^ itro) -phenylisoxazol-5-yl-acetyl chloride (prepared from 1.64 g (6.6 mmol) of the corresponding acetic acid and thionyl chloride) with N, 0-Bia -trimethyleilyl-7-aia.Lnocephalosporanat (prepared in situ from 1.8 g (6.6 mmol) of 7-aminocephalosporanic acid) converted. The reaction product was worked up in the usual way. Was isolated in 1350 mg (40 i ') of a schv / achgelb stained Pegtstoffes · Gernäß DüpnschicKtchroniatographiej of the IR and NMR spectra of the final product, the purity was about 85 ^c />.

The analysis of the HIIR spectrum of the end product, dissolved in hexadeuterodimethyl sulfoxide (60 Mc ₁ (/ '- words in ppm, internal standard 2,2-diinothylsilapentane-5-sulfonate) gave the following values:

9.36 and 9.22 (J = 8.0 ± 0.5 cpn, about 0.8 protons)

7> 9 - * 0.5 (AA'DB splitting pattern, 4 protons)

209820/1159

BAD ORIrtlNAI Isoxazolyl C ₄ -H 7.05 (1 proton)

C ₇ -H 5.87, 5.79, 5.73 and 5.65

(weakly broadened signals,

Jsy 8.0 cps and J ^ _B <= ⁱ 4.6 cps, 1 proton)

C ₆ -H 5.19 and 5.11 (J ^ => 4.6 t 0.2 cps)

₂ (5.19), 4.97, 4.82 and 4.60 C \

( ^J AB ^{s 13} »° - ° ' ^{2 cps)}

CH ₂ -CO at 4.0 (slightly broadened singlet,

2 protons)

S-CHp at 3.6 (center of an AB quartet with

very faint outer lines, 2 protons)

CO-OH, 2.06 (3 protons.

Example 25

A quantity of 100 to 2000 mg of the sodium salt of 7 ~ £ - [3- (2,6-dichlorophenyl) -i8exuz.ol-5- J i! -Acetamido} -cephalosporanic acid is aseptically converted into a composition suitable for injectable Ampoule inserted. Before use, the powder is dissolved in a suitable amount of sterile and pyrogen- free water.

209820/1159

BATH ORIGINAL

Example 26

The compounds of Examples 1 to 20 became syrups made by mixing the following ingredients:

Sodium salt of the compound used 1.5 - 6 β

Loessllohe strength 1 -5g

Sodium cacharine 0.1-1 g

Hipa H 0.06 g

Strawberry flavor 0.1 - 5 6

Amaranth 0.010 g

Sucrose 30 g

V / aaser to a volume of 60 ml «

These syrups are suitable for oral administration *

Example 27

Capsules are commonly manufactured to be active Active ingredient contained a compound according to Examples 1 to 20 «They components of the capsules are the following:

Hatriumealz of the compound used 150-500 mg Potassium bicarbonate 100-300 mg

MagneaiUEiBtearate 2-10 rag

Lactose (sufficient for 1 capsule)

These capsules can be administered orally

209820/1159

215508t

Example, 28

In the usual way, tablets were produced which, as an active ingredient, contain a compound according to Examples 1
to 20 contained.

The ingredients of the tablets are as follows:

Sodium salt of the compound used 125-1300 mg, polyvinylpyrrolidone 5-30 mg

Amyluia-Ilais 100-300 mg

Magnesium acid content 1-2C mg

Lactose (sufficient for 1 tablet)

These tablets can be administered orally.

209820/1159

Claims (1)

PATENT CLAIMS: Compounds of the general formula N. O - C: ι » R ² ° N-Q (D wherein Q is a group of the formulas H
CC -C - Ν - H - C or I. 'COOH (II) CH, C - CH ₀ X C. COOH (III) represents, v / orin X is a vaco carbon atom, a hydroxyl group or a Niodri ^ .Aliumoylgruppe means, or the group -C 'L, vC' and the carboxyl group of Porw ·)! I'll be taking education a Laclon group of the formula 209820/1159 BATH ORIGINAL -70-215508t -CO-CH ₀ -0 or a lactam group of the formula -C-NH-CH ₀ -S are connected, H is a lower alkyl or an aryl, such as a phenyl or a haphthyl group, which optionally has 1 or more substituents, such as chlorine atoms, fluorine atoms, nitro groups, amino groups or lower alkyl groups, or the 9,6-Diehlophenylgruppe or R a tertiary Alkyl group, such as an adamantyl group, R represents a hydrogen atom or a lower alkyl group; - »e or an optionally esterified with a lower alkyl group or into an alkali metal salt, an alkaline earth metal salt or an amine as converted carboxyl group, a Cr-rbamylgruppo, a cyano group, an amino group or a chlorine atom, ji a '., Jo, oxygen atom, a cyano group, an amino group, a lower alkoxycarbonylamino group, a lower alkyl group, a carboxyl group which is substituted with a diodoxy group such as a phenyl group or an aralkyl group such as a benzyl group, or a carbanoyl group which is optionally bears a lower alkyl or phenyl substituent on the nitrogen atom, and the alkali metal salts, the alkaline earth metal salts and the amine salts, the esters, such as the tri- (NiGiirin) -nlkylrsi.lyßter, the di- (lower) -alkyl-monohalogcnailyl esters _t the The phenacyl or the amides, ie the saccharide derivatives of the pericillanic acids and cophalonic acids mentioned. 2, compounds according to claim 1, characterized in fjOJionn ;; oich ~ net, dai3 the group R in formula 1 c: Lno foothyl-, Üthyl-, 209820/1159 β ± »β gebeaenfüElS 'ilfci? o & an offer several aubstituen-th _t -like Ghioratoae, fluoimtoae. Hitro groups, amino groups, phenyl groups or ethyl groups substituted by phenyl groups, or a teirfeiäre A Hey! Group, such as an adainantyl group, R a hydrogen atom, a methyl, ethyl, propyl, butyl group or a carboxyl group, which may optionally be substituted with a methyl group. , Propyl or butyl group- esterified or into a sodium, EaliuDä, calcium or amine salt overrun, or H 2 a Car bar., R, Gyano group ether a Clilor atom and E öin WOifatoBi water, a Gyano- *, amino en B sy Iq xy car bony 1-amino- _t Nethy1-, ethyl, propyl or butyl group, with a iiethyl-, ethyl, ^ _r ropyl- butyl, phenyl - or Seazvj.gruppe esterified carboxyl group or a carbamoyl which optionally bears a methyl, propyl, butyl or phenyl substituent on the nitrogen atom, as well as dJLe Hatriuin- _r potassium, ftalsiuia- and amine salts, the $ rii3ethyloilyl ~ _r Dime thy liaonohalogensilyl-, triethylsilyl-, diethyiuionobii.lc _c -nsilyl-, benzyl- and phenacyl esters and the saccharinyl-, succiniaido- or phthalimidoiraitl-derivatives of these penicillan and ceplialosporan acids * 3 * Compounds according to claim 1, characterized in that II the 2,6-l) chlorophores: yl or the 2,6-chloro flvorpliony If luppe, R a Uasßeriitoffatojn he locates a diethyl p
group, Ii a Vasn ^ rntiuTatGiB od <.3 * oine Ik, Uiy! group and Q a GephaTohpoi ^ inrjawrcroct boticutcn, i, ou.io diG brawn diesvx * "V 4. 6-t- [3 - (? Fi * »- 3> Jp
anido] -ponici3] li * Hr iurc υηο ΰ (· ι · υιι 5. 7- {- [3-C 2,6-dicilophenyl) -isoxazol-5-yl j -acetaaido} cephalosporanic acid and their salts. 6. ή-Ι- [3- (2,6-Bichlorphenyl) -isoxazol-5-yl 3-acetamido3-deacetoxyoephalosporansawre and their salts. 7. 6- 1- [3- (2, 4, 6-trimethyl) phenyl isoxazol-5-yl] acetamido] penicillic acid and salts thereof. 8. 7-C- [3- (2,4 »6-trimethyl) -phenyl-isoxazol-5-yl] -acetamido} -cephalosporanic acid and their salts. 9. 7-i- [3- (2,6-dicbloro) -phenyl-4-ethyli-soxazol-5-yl3-acetamidol-deacetoxycephalosporanic acid and their salts 10. 6-fcx ~ chloro [3- (2,4,6-trimethyl) phenyliso3cazol-5-yl3-acetamido] ~ penicillanic acid and their salts. 11. 6- {~ [3- £ ^, e-Di acetamidoj ~ penicillanic acid and its salts. 12. 6- {α-Methyl [3- (2,4,6-trinetyl) -phenyl-isoxaeol-5-ylj-acetamidol-penicillanic acid and their salts 13. 7-2- [3- (2,4 _f 6-Triraethyl) -phenyl-4-r-diethyl-isoxazol-5-ylI-acetamidoJ-cephalosporanic acid and its salts. 14. 7- (- [3- (2 ρ 4,6-Tricfethyl) -fhettyl - ^ - mef üylissOxazol-5-ylj-acetamidoj-desacetoxycephaloeporanoic acid and its salts. 15 · 6 ~ {~ [; 5- (2,6 ~ dichloro) ~ phenyl ~ 4-oarbamyli.30xasol ~ 5-yl} -acetamidoj-penieillanic acid and their Sa.lse. 209820/1159 2T55081 16. 6-ί- [3- (2,6-DietLtor) -phenyl ^ -cyano-isoxazol-S-yl! -Acetamido] -penicillanic acid and their salts. 17 · 6- £ - [3- {1) -Adamantyl-ieoxazal ~! 5-yl! -Acetamido} penicillanic acid and its salts, 18 · 6 - {<t- (p-Witro) * - benzyloxycarbonylamino-C3- (2,6-dichloro) ~ phenylisoxazol-5-yl! -Acetamido} -penicillanic acid and its salts " 19. 6- oC-Amino- [3- (2,6-dichloro) -phonyl-isoxazol-5-yl} -acetamido -ponicillanic acid and its salts. 20. 6-i- [3- (4-nitro) -phenylisoxazol-5-yl! -Acetamido} penicillanic acid and their salts. 21. 6- {ct -carbamyl- [3- (2,6-dichloro) -phenyl-i-doxazol-5-ylI-acetaiaidoj -penicillapsic acid and its sal ^ j. 22. 6-i- (3-Methyl-isoxazol-5-yl) -acetamido3-penicillanic acid and their salts. 23. 7-i- [3-Hothyl-isoxazol-5-yl] -ocetaraidoj-penicillanic acid and their salts. 24. 6-i ^ - (N-phenyl) -carbarayl- [3- (2,6-dich.lor) -phenyli8oxazol-5-yl] -acetamidoi-penicillanic acid and their salts. 25. 7-ί- [3- (4-17itro) -phenylisoxazol ~ 5-yl! -Acetamido] - · ccphulospornic acid and its brawn * 209820/1169 26, process for the preparation of the compounds according to Claim 1, characterized in that there is a courtship, a Ester or an aide of an S-aminopenicillanic acid or 7-aminocephalosporanic acid compound of general formulas HH s ^ ^CII 3 CN C ^or / CN .C- CII ₉ X Κ / ^N COOH O ^ \ ^ * ι COOH before X has the meaning given in claim 1, where the substituent X, when it represents a hydroxyl group, is preferably protected with an active ester of an acid of the general formula RR ¹ II
-C- COOH R ² < ^VI > 1 2
wherein E, R and R are as indicated in claim 1 »Γ> ου αί own, or a functional derivative that al ::; Acylating agents for a primary amino group can act in an 8i? .H b ejcann b er V / eise um «ο t ζ t. 27 · Pharmaceutical Additions to _£ " ₍ in, dnih!, ·: · '» F; ek-. «· .: indicates that they contain one or more compounds according to claims 1 to 25 as well as a suitable pharmaceutical 209820 / TT5'9 BAD ORlQINAL Träglicliee carrier material included. 28. T compounds have the general formula odei their derivatives, wherein R is a Niedri ^ alkyl group, a Aryl group, such as a phenyl group or an 'iaphtbyl group, which optionally have one or more substitutes, such as Chlorine atoms, fluorine atoms, nitro groups, amino groups, lower alkyl groups wear, such as the 2,6-dichlorophenyl group or a tertiary alkyl group such as the adamantyl group, R a 1 iedri £? Lkyl ~, carbamyl, cyano, amino group or a ρ
Chlorine atom, R a cyano, amino, aralkoxyearbonylamino, lower alkyl or carbamoyl group, which can optionally carry a lower alkyl or phenyl substituent on the nitrogen atom, and Y a carboxyl group, which can optionally be converted into an alkali metal salt, an alkaline earth metal or an amine salt or an ester is converted, or represents a group He or Mc-Hal, in which Me represents a metal atom, Kummer I or II represents the valency of this metal atom and Hal represents a halogen atom, such as a chloratoin or a bromine atom. 29. [> - (1) -Adaraantylisoxazol-! 5-yl] -essigsili're and their Salts and esters. 209820/1159 BATH ORIGINAL 30 · 13- (4-Hitro) -piieiijliÄ -oxazol-5-ylJ-acetic acid «aid their salts trad ester * 31. [3 * C4-Afflino) - piieöjlisoa3aa5ol-5-jl3-ee «igsäMre and their salts and esters. 52. <s ~ Broa-C 5-i 2 _f 6-dI «3ilor) -pheny lisoxazol-5-y 13- acetic acid and its salts and esters, 33. ο, imino «[3- (2,6-dieiaor3 i3äiea ~ * ~ yli6omzol 5-yl 3-essigeäiare« nd their salts w & ä ester, 34 · f 3- (2 _f 6-J) ic3ilor) ^ phen3rl-4 ~ ffletli3rlisoxaaol ~ 5-yl I-acetic acid and its salts and esters, 35. α ^ chloro) -pbenylisoxazole ~ 5-Tl3-ßbic acid and its salts and Ester. 36 »ί 3- {2 _f 6-Diciilor3-plienyl-4-« yaiao-isoxazol-5-yl 3- «Acetic acid and its salts and esters, acetic acid and its salts and esters 38, £ 3- ί 2 f 4,6-friaethyl) -phetiyl - 4-HBielByli © O2ia E.ol-5-yl 3- acetic acid vlivJ its salts uM Bster, 39 · o-Methyl- [3- < 2,4,6-trinethyl) -pbenylisoxazol-5 ~ yl 3- acetic acid and its salts and Eat-ur, 209820/1159 40. oc-chloro-C3- (2,4f 6-trimethyl) -phenylisoxazol-5-ylJ-acetic acid and their salts and esters. 4L Process for the preparation of substituted isoxazol-5-yl-acetic acids and their derivatives of the general formula RR ¹ - CH - COOH H ² wherein R is a lower alkyl group, an aryl group such as a phenyl group or a naphthyl group, which optionally by one or more substituents, such as chlorine atoms, flua: at '). ae, nitro groups, amino groups, lower alkyl groups may be substituted, such as the 2,6-dichlorophenyl group or a tertiary alkyl group such as an adamantyl group, R is a lower alkyl group, a carbamyl group, a cyano p group, an amino group or a chloratora and R a Cyano group, an amino group, an aralkoxycarbonylaiaino group, a lower alkyl group or a carbamoyl group, which may optionally have a lower alkyl or phenyl substituent on the nitrogen atom, are characterized in that a compound of the formula RC-Ii -> 0 with a compound of the general formula R -G = G-GU ₀ -Z ₁ 1
in which R and R have the meanings given above and Z is a hydrogen atom, a carboxyl group, ei! io Xtthy1 group or a hydroxyl group, is subjected to a cycloaddition reaction, then, if appropriate, a reaction to change the group R is carried out, one 209820/1159 BATH ORIGINAL when Z is a hydrogen atom or a methyl group, to introduce a carboxyl group into the compound thus obtained, this connection with a Lithiujaalky! connection and COp and H ^ O converts, or the compound obtained, if Z denotes a hydroxyl group, oxidizes and the compound thus obtained is further oxidized to achieve a ct-llalogenation reacted, whereupon the compound thus obtained is treated with nucleophilic agents or with a lithium alkyl compound and a suitable means and, if necessary the acids obtained are converted into their salts or esters. 209820/1159 BATH ORIGINAL